front page 08.BMP - Pakistan Journal of Entomology Karachi

Transcription

Pakistan j. entomol. Karachi. 23 (1&2): 2008
CONTENTS
DETERMINATION OF MAMMALIANS TOXICITY OF CEDRUS DEODARA ROOT OIL,
AGAINST ALBINO RATS (WISTAR STRAIN).
PERVEEN, R., NAQVI, S.N.H., M. AHMED AZMI, AHMED, M. & SAIMA
MEHMOOD………………………………………………………………………………………........
01-04
TAXONOMIC STUDIES OF SERGENTOMYIA FREETOWNENSIS SINTON, 1930,
SINDHICUS N. SUB. SP. (DIPTERA: PSYCHODIDAE) IN SINDH, PAKISTAN.
JUMA KHAN KAKARSULEMANKHEL……………………………………………………………..
05-10
COMPARATIVE TOXICITY OF ORGANIC SOLVENTS AND INSECTICIDES AGAINST
CABBAGE BUTTERFLY.
SEEMA TAHIR, TAHIR ANWAR, MUHAMMAD SAMIULLAH CHANNA, IMTIAZ AHMAD &
YOUSEF HAYAT KHAN……………………………………………………………………………..
REDESCRIPTION OF CAYSTRUS PYGMEAUS LINNAVUORI
PENTATOMIDAE) AN ISOLATED SPECIES FROM ETHIOPIAN REGION.
11-14
(HEMIPTERA:
MUHAMMAD ZAHID & IMTIAZ AHMAD…………………………………………………………...
STUDY OF RESISTANCE IN SITOPHILUS ORYZAE AGAINST
CYPERMETHRIN AND PHOSPHINE ON THE BASIS OF TOXICITY VALUES.
15-17
BIOSAL,
S.M. NAUSHAD ZAFAR, NAQVI, S.N.H., TARIQ, R.M. & REHANA PERVEEN ……………..
19-26
EFFICACY OF IMIDACLOPRID AND ENDOSULFAN IN COMPARISON WITH BIOSAL
(BIOPESTICIDE) AGAINST MYZUS PERSICAE (SULZER) ON MUSTARD CROP.
MUHAMMAD FAHEEM AKBAR, NIKHAT YASMIN, FARAH NAZ & NAZIA
QURESHI………………………………………………………………………………………………
27-30
REVISION OF THE GENUS ISOMETRUS HEMPRICH & EHRENBERG (SCORPIONIDA:
BUTHIDAE: CENTRURINAE) WITH DESCRIPTION OF TWO NEW SPECIES AND
CLADISTIC RELATIONSHIP FROM PAKISTAN.
RAFAT AMIR & SYED KAMALUDDIN …………….. ……………………………………………..
31-40
TAXONOMIC STUDIES OF SERGENTOMYIA BAGHDADIS (ADLER & THEODOR) IN
SINDH AND PUNJAB AND ITS PHYLOGENETIC RELATIONSHIPS.
JUMA KHAN KAKARSULEMANKHEL …………………………………………………………….
41-46
POTENTIAL OF SWEET FLAG RHIZOME OIL AND CUSTARD APPLE SEED OIL
AGAINST THE MAJOR SUCKING PESTS OF COTTON, AS COMPARED WITH
CONFIDOR + DELTAPHOS, AT ARI-TANDOJAM, SINDH-PAKISTAN.
RAJPUT MUHAMMED TARIQ, S. NAIMUL HASAN NAQVI, S.M. NAUSHAD ZAFAR &
ABDUL SATTAR BURRERO………………………………………………………………………..
47-50
STUDIES ON SAMPLING TECHNIQUES TO MONITOR ADULT POPULATION OF
WHITEFLY, BEMISIA TABACI (GENN.) IN CUCURBIT FIELD.
ABDUL GHANI LANJAR, MUHAMMED KHAN LOHAR, HAKIM ALI SAHITO, ASHFAQUE
AHMED NAHIYOON AND NAHEED BALOCH……………………………………………………
51-54
POPULATION DYNAMICS OF GROUND WATER BREEDING OF CULEX MOSQUITOES
OF KARACHI AND THATTA DISTRICT.
TANVEER FATIMA SIDDIQUI……………...............................................................................
LEVELS OF DENGUE FEVER VIRUS CONTROL: THE FFECTIVENESS AND VASTNESS
OF CONTROLLING POWER BOUNDARIES OF THESE LEVELS
RAJPUT MUHAMMED TARIQ & S. SALAHUDDIN QADRI……………………………………..
55-60
61-62
ENTOMOLOGICAL SOCIETY OF KARACHI, PAKISTAN (1971)
Office Bearers and Council for the year 2008-2009
President
S.N.H. Naqvi: D.Sc.: HEC Eminent Professor, Department of Zoology, University of
Karachi, Gulshan-e-Iqbal Town, Karachi-Sindh.
Vice-Presidents
Abdul Sattar Burrero: Ph.D. (Sindh), Director General, Agricultural Research
Institute, Tandojam, Sindh.
General Secretary
M. Arshad Azmi: Professor, Department of Zoology, University of Karachi, Karachi.
Joint Secretary
Tahir Anwer: Incharge, Pesticide Research Institute, SARC (PARC), University
Campus, Karachi.
Treasurer
Masarrat J. Yousuf: Professor, Department of Zoology, University of Karachi,
Karachi.
Councilors
S. Amin ullah Khan: Ph.D., M. Ahmed Azmi: Ph.D., Rukhsana Perveen: Ph.D.,
Rahila Tabassum: Ph.D., M.S. Wagan: Ph.D., Nasreen Memon: Ph.D., Muhammed
Zahid: Ph.D., S. Salahuddin Qadri: Ph.D.
Entomological Society of Karachi was established in 1971, with the object of
promoting Entomological Science and a closer cooperation between entomologists
of Pakistan. Pakistan J. Entomol. Karachi (Biannual) is available in exchange or by
subscribing the cost, from the Society’s office at the Department of ZoologyEntomology, University of Karachi, Karachi-75270, Pakistan.
The Journal is being abstracted by Biological Abstracts, Entomological
Abstracts, Chemical Abstracts and Current Contents and recognized by HEC in
“Y” category.
Due to high cost of publication the page charges are Rs.2500.00 for 1-4
pages and Rs.500.00 per page for extra pages or US $ 10.00 per page from
members and Rs.700.00 per page or US $ 15.00 per page from non-members.
Subscription Price
Pakistan
Rs.400.00 per copy/number
Other Countries
US $ 40.00 per copy/number
Composed and Designed at Muhammed Afzal Husain Qadri Biological Research
Centre, University of Karachi, Gulshan-e-Iqbal Town, Karachi-75270, Sindh-Pakistan
Pakistan J. Entomol. Karachi, 23 (1&2): 2008
Foreign & National Editorial Board
Carl Schaefer, Ph.D.
Michael Breuer, Ph.D.
University of Connecticut, Storrs, Conn. (USA)
Zoological Institute, Catholic
University of Leuven (Belgium).
A.R. Shakoori, Ph.D.
Kehkashan Akhter, Ph.D.
University of the Punjab, New Campus, Lahore
(Pakistan)
Department of Zoology, University of Karachi,
Karachi-75270, Sindh-Pakistan.
M.F. Khan, Ph.D.
M.A. Matin, Ph.D.
Department of Zoology, University of Karachi.
National Agricultural Research Centre (NARC),
Park Road, PO NIH, Islamabad (Pakistan).
S. Anser Rizvi, Ph.D.
R.C. Saxena, Ph.D.
Chairman, Neem Foundation, Mumbai, India.
Feyzi Onder, Ph.D.
M. Ather Rafi, Ph.D.
Department of Plant Protection, Agric. Faculty,
Ege Univ. Bornova, Azmir (Turkey).
National Agricultural Research Centre (NARC),
Park Road, PO NIH, Islamabad (Pakistan).
Seema Tahir, Ph.D.
Dr. Jumakhan Kakarsulemankhel, Ph.D.
Department of Zoology, University of Balochitan,
Saryab Road, Quetta, Pakistan.
Foreign Advisory Board
U.S.A.
Asia
Alfred Wheeler Jr. Ph.D.
Cornell University, Itacha, N.Y.
Chiu, Shin-Foon, Ph.D.
South China Agriculture Guangzhou
(Peoples Republic of China)
T.J. Henry, Ph.D.
US National History Museum
Washington, D.C. (USA)
V.K. Ganesalingam, Ph.D.
University of Jaffna
(Sri Lanka)
J. Koolman, Ph.D.
Philips Universitat
Marburg (Germany)
B.N. Islam, Ph.D.
BAU, Mymensingh, University
(Bangladesh)
R.W. Mwangi, Ph.D.
University of Nairobi
P.O. Box 72913, Nairobi (Kenya)
K. Sombatsiri, Ph.D.
Karetsart University
Bangkok (Thailand)
J.I. Olaifa, Ph.D.
University of Technology
Ogbomoso (Nigeria)
R.P. Singh, Ph.D.
Entomology Div., IARI
New Delhi 10013 (India)
Errol Hasan, Ph.D.
University of Queensland,
Gattons College, Lawes, QLD.
Absar Mustafa Khan, Ph.D.
Department of Zoology
M.U. Aligarh (India)
Europe
Africa
Australia
INSTRUCTIONS TO CONTRIBUTORS
(For Research and Review Articles)
1.
Manuscripts should be in English, typed and double spaced, on one side of the
paper. There should be an abstract, not exceeding 200 words, which will be
printed in small type before the introduction. Nothing in the text, except the
scientific name will be italicized. Tables should be typed on separate sheets.
Footnotes should be avoided as much as possible.
2. Illustrations should be in black Indian ink preferably on white or transparent
paper. All letterings and numericals should be in light pencil for insertion in
uniform style. The author’s name and the number of the figures should be
written on the back of each drawing. The size of the illustration after reduction
will not exceed 4x6 inch. The legends should be typed on a separate piece of
paper.
3. The author should submit a hard copy in original and a photocopy of the original
and soft copy in re-writable C.D.
4. Photographs should be glossy black and white prints. They should be
numbered separately from the Text-figures. The cost of the coloured plates will
be met by the author.
5. Reference should be cited in the text by giving the author’s name followed by
the year. Papers and books should be listed in the alphabetical order. Journals
should be abbreviated according to the latest edition of the World List of
Scientific Periodicals e.g.
DREYER, M. (1984). Effects of aqueous neem extracts and nee mol on the
main pests of Cucurbita pepo in Togo. Proc. 2nd Int. Neem Conf.
(Rauischholzhausen, 1983), pp. 435-443.
EDWARDS, C.A. AND HEATH, G.W. (1964). The Principles of Agricultural
Entomology. Chapman and Hall, London, 418 pp.
NAQVI, S.N.H., ASHRAFI, S.H. AND QADRI, M.A.H. (1968). Acid phosphatase
activity in the digestive system of the desert locust, Schistocerca gregaria
(Forskål). Aust. J. Biol. Sci. 21: 1047-52.
RAUPP, M.J. AND DENNO, R.F. (1983). Leaf age as a predictor of herbivore
distribution and abundance. In: Denno, R.F. and McClure, M.S. (eds.): Variable
Plants and Herbivores in Natural and Managed Systems. Academic Press, New
York, USA, pp. 91-124.
6. Page-proofs will be sent to the authors for correction which should be returned
within 7 days. Authors may be required to pay for alternations in proofs other
than those needed to correct printer’s errors. The responsibility of the results
will be on authors(s).
7. Key words/Index of maximum 10 words and short title of 8 words be mentioned
at the proper place.
8. Authors may order for purchase of extra reprints when proofs are returned to
the Editor. After acceptance of paper the amount of publication charges should
be paid before publication of the paper.
9. All correspondence should be addressed to the Executive Editor, C/o Office of
the Entomological Society of Karachi, Department of Zoology, University of
Karachi, Karachi-75270, Pakistan, or Assistant Editor, MAH Qadri Biological
Research Centre, University of Karachi.
10. Paper(s) for publication may be emailed at [email protected] or
naeemnaqvi.life.com.
Pakistan j. entomol. Karachi 23(1&2): 1-4, 2008
DETERMINATION OF MAMMALIAN TOXICITY OF CEDRUS DEODARA
ROOT OIL, AGAINST ALBINO RATS (WISTAR STRAIN)
PERVEEN1, R., NAQVI1, S.N.H., M. AHMED AZMI1, TARIQ2, R.M.,
AHMED3, M AND MEHMOOD1, S.
1
Baqai Institute of Pharmaceutical Sciences, Baqai Medical University, Super Highway, Karachi.
2
M.A.H. Qadri Biological Research Centre, University of Karachi, Karachi-Sindh, Pakistan.
3
Aga Khan Medical University, Stadium Road, Karachi.
ABSTRACT
Mostly Cedrus deodara root oil has been analyzed for sesquiterpenes and hydrocarbons, However, in
the present case GC-Mass analysis and spectral studies were done on root oil. This oil is being used
orally as anti-ulcer agent by Hakeems. However, scientifically it is necessary that the toxicity level be
determined against mammals. During present investigations mammalian toxicity was determined, by
oral administration, against albino rats (Wister strain). The LD50 by probit mortality graph was found to
be 34.4 gm/kg. This is quite safe as compared to Neem oil LD50 (5gm/kg).
Key words: Mammalian toxicity, Cedrus deodara root oil, albino rats.
INTRODUCTION
Essential oils are volatile and hydrophobic. They
are used in various industries, household cosmetics
and medicine etc. The production of Cedrus deodara
oil is about 2600 tons per year. The oral LD50 of
various essential oils various from 0.5-5gm/kg
(Tisser and Robert 1995).
According to MSDS (Material Safety Data Sheet)
reported by Environmental Health and Safety
Department, USA. Cedar wood oil may cause
irritation to skin, eyes and respiratory tract (Case No.
8000-27-9). The oral LD50 for rat is 5gm/kg, for rabbit
skin 5gm/kg and for draize test 500mg (MSDS)
(Anonymous 2003) GC mass analysis of oil showed
that C. deodara root oil contains deodarone and
atlantone (Sankarananyan et al., 1973).
Agarwal and Rastogi (1981) reported presence
of himachalol, himasecollone and cendarol in trunk
oil. Avcibasi et al. (1987) reported four terpenoids
i.e. α torosol, βtorosol, andirolactone and
transatlantone. Khan and Naheed (1990) reported a
new sesquiterpene, himachanene from bark of
Cedrus deodara. Molluscicidal activity of cedar wood
oil, neem oil was reported by Rao and Singh (2001).
Dimiri and Sharma (2004) reported acaricadal
effect of Cedrus deodara oil, Pomgamnia glabra oil,
Jatropa curcus oil and Benzyle benzoate, on sheep
mange mite. This indicates that these oils though not
toxic to mammals but are toxic to insect and
acarines and control the ectoparasites.
As cedar wood root oil was being used for
healing peptic ulcer in North Western Province of
Pakistan, therefore, it was necessary to find the
mammalian toxicity of this oil, which will help in
adjusting the therapeutic dose. For this reason LD50
of the root oil was determined.
MATERIALS AND METHOD
For toxicity determination albino rats (Wistar
strain) were taken. Rats were starved for 12 hours.
The weight of the rats was 250gm ± 5gm. Ten rats
were not treated (control) while 5 batches of 10 rats
each (5 male+5 female) were treated with oil. Then
2.2ml, 4.4ml, 6.6ml, 8.8ml and 11.1ml, oral dose was
given orally by special gauze syringe. After 24 hours
mortality readings were taken.
The volume was converted to grams by
multiplying volume with specific gravity of oil (0.91).
Five replications were done and readings were
subjected to Abbot's formula (1925). The rats were
allowed to feed on normal diet after experiments.
Experiments were conducted under controlled
condition at a temperature of 25°C + 2 and.relative
humidity as 50 + 2. The probit morality graph was
drawn and LD50 was noted.
Perveen et al.
2
RESULTS
Whenever, a new compound or drug is to be
used for clinical purpose, it is necessary that toxic
dose for mammals be determined. During present
experiments, the toxic dose of Cedrus deodara root
oil was determined, as it is being used as anti ulcer
drug in NWFP by Hakeems.
Toxicity experiments were done under controlled
conditions as mentioned above, on standard Wistar
Strain, being breed in animal house of Baqai Medical
University. The rats were treated with 2.2, 4.4, 6.6,
8.8 and 11.1 ml dose/rat. The average mortality after
24 hours was found to be 10, 22, 40, 49 and 68%
respectively. In the control batch (untreated) no
mortality was recorded i.e. zero.
According to the graph the LD50 was found to be
34.4gm/kg + 1.2 gm (Fig. 1) standard deviation, and
range at 95% confidence limit was also determined
(Table 1).
DISCUSSION
When a plant product is found effective against
any disease/disorder. The product acquires the
status of medical importance and termed as
medicinal plant. There are some barriers for using
and commercialization of these type of plant
products as reported by Isman (1997). This is
because of its acute toxicity, piscean toxicity, as
reported by Tariq et al. (2004), avian toxicity as
reported by Tariq & Naqvi (2006) and mammalian
toxicity as reported by Naqvi & Tariq (2007).
After the satisfaction or low toxicity of the
candidate plant product, it is tested against the small
mammals e.g. on rats, mice and rabbits before going
on direct application to human beings as Shirwaikar
et al. (2004) reported anti-diabetic activity of
aqueous leaf extract of Annona squamosa in
streptozotocin-nicotinamide type 2 diabetic rats.
Therefore in the present work, root oil of C. deodara
was tested against rats, as the oil acts as anti-ulcer
in human beings.
The LD50 (34.4 gm/kg) of the cedar wood root oil
is very safe. According to the classification of
Casarette and Doull (1992) any dose above 15gm/kg
is very safe. Most of the essential oils, cotton seed
oil, sunflower oil, clove oil, corn oil etc. are very safe
or approximately non-toxic. The LD50 of there oils
varies from 4-5 gm/kg or more (Tisser and Robert
1995). The oral LD50 of cedar wood oil (red juniper)
has been reported as 5 gm /kg. However, in the
present case the root oil of Cedrus deodara is much
safer than juniper oil. (MSDS) (Material and Safety
Data Sheet) issued by Department of Environmental
Health and Safety, USA.
The oral LD50 of neem oil has been reported as
4.2 gm/kg which is close to the LD50 of red cedar
wood oil, but this is also in safe category. Acute
toxicity of neem oil has reen reported as 3540mg/kg
to 5000 mg/kg (Farm Chem. Hand book 1995), and
US EPA (1993). During present analysis of root oil
three compounds were isolated viz, himachalol,
allchimachalol and trans-atlantone which are already
reported from trunk oil (Perveen 2006).
Toxic/ therapeutic affect of crude oil and isolated
compounds were also observed which have been
reported elsewhere. It has been found that the
Cedrus deodara root oil is quite safe and may be
used for therapeutic use and pest control as reported
by Dimiri and Sharma (2004). However, some mild
pathological effects were observed during present
investigation.
Naqvi & Tariq (2007) reported the mammalian
toxicity of two medicinal local plants, Acorus calamus
and Annona squamosa, against white rats. They
reported 8.7 ml = 38.37 gm/kg LD50 of calamus oil.
Whereas the LD50 of squamosa was reported as 7.7
ml = 33.95 gm/kg against the same wistar strain of
albino rats. The LD50 value of squamosa is very
close to the LD50 value of deodara in present
findings. Whereas the LD50 of calamus is more lower
as compared to deodara, being more safer than
deodara, but the LD50 of three local plant falls within
the safe limits that is above 15 gm/kg. Therefore the
root oil of C. deodara may be used safely as antiulcer.
CONCLUSION
The LD50 dose of C. deodara against small
mammal, Wistar strain (Albino rats) has been found
34.4 gm/kg, which is above 15 gm/kg dose.
Therefore it will also be safe for human beings as
well and may be used safely as anti-ulcer as
concluded experimentally in the present work.
Beside this, the oil of Acorus calamus which is also
used as medicine for stomach complaints, snake
bite, for remittent fevers Nasir (1978), and insect
repellent Tariq & Qadri (2001). The Acorus is locally
found in Pakistan and may be experimented for
above disorders in human beings. Further more the
leaves of Annona squamosa are reported having
anti-diabetic activity, by Shirwaikar. This plant is also
frequently available in Pakistan especially in Karachi.
It may also be taken into practical use after research
as the diabetic problem is a major problem in
Pakistan. The main active ingredient of Acorus oil is
the β-asarone. Streloke et al. (1989) and neoanonin
(squamosin) is the main active ingredient in
squamosa seed oil. Kawazu et al. (1989).
3
Determination of Mammalian Toxicity of Cedrus deodara root oil
Table 1
DETERMINATION OF TOXICITY OF CEDRUS DEODARA OIL AGAINST ALBINO RATS
Obs. No.
Dose per rat (ml)
Mean % mortality
S.D. +
S.E. +
Range at 95% confidence limit
0
Control
Zero
Zero
Zero
-
1
2.2
10
4.4
2.01
6.2-14.35
2
4.4
22
4.47
2.20
18.08-25.90
3
6.6
40
6.7
3.02
33.80-45.19
4
8.8
49
7.01
3.16
43.70-55.20
5
11.1
68
7.20
3.12
61.10-74.90
N = 5 range = mean + S.E. x significant value at 95% confidence limit.
Perveen et al.
4
REFERENCES
ABBOT W.S. (1925). A method of computing
effectiveness of an insecticide. J. Econ.
Entomol. 18:265-67.
AGARWAL, P.K. AND RASTOGI R.P (1981).
Terpenoids
from
Cedrus
deodara.
Phytochemistry 20(6); 1319-21
ANONYMOUS (2003). Material Safety Data Sheet
issued by Environmental Health and Safety
Department, USA.
AVCIBASI, H., ANIT, H. AND TOPRAK, M. (1987).
Four ter-penoids from Cedrus libanotica.
Phytochemistry 26(10): 2852-54.
CASARETT, L.J. AND DOULL, M.D. (1992).
Toxicological evaluation: The Basic Science of
Poisons. Published by Macmillan Publishing Co.
Inc. New York. Pp. 11-25.
DIMIRI, V. AND SHARMA, M.C. (2004). Effects of
Sarcoptic mange mite and its control with oil of
Cedrus deodara, Pangamia glabra, Jatropa
curcu’s and Benzyl Benzoate, both with and
without ascorbic acid on growing sheep,
assessment of weight gain, liver function,
nutrient digestion ability, wool production and
meat quality. J. Vet. Med. A (Physiol. Pathol:
Chem. Med) 51 (2): 79 - 84.
FARM CHEMICALS HANDBOOK (1995), Meister,
Publishing Co. Willough by, OH.
ISMAN, M.B. (1997). Neem and other botanical
insecticides: Barriers to commercialization.
Phytoparasitica 25(4): 339-344.
KAWAZU, KAZUYOSHI, JOCELYN, P., ALCANTARA
AND KOBAYASHI, A. (1989). Isolation and structure
of neoanonin, a novel insecticidal compound from the
seeds of Annona squamosa. AGRIC BIOL CHEM
53(10): 2719-2722.
KHAN, K., AND NAHEED, S. (1990). Chemical
investigations of Cedrus deodara. Stem bark
Isolation
and
identification
of
some
sesquiterpene hydrocarbons. J. Chem. Soc.
Pak. 12(4): 282-84.
NAQVI, S.N.H. AND TARIQ, R.M. (2007).
Mammalian toxicity of Acorus calamus (AC) and
Annona squamosa (AS) oil against Albino rats.
Pakistan j. entomol. Karachi. 22 (1&2): 37-38.
NASIR, Y.J. (1978). Araceae. Flora of Pakistan N o.
120. In E. Nasir and S.I. Ali (eds). Department of
Botany, University of Karachi and National
Herbarium, Agricultural Research Council,
Islamabad.
PERVEEN, R. (2006). Analysis of Cedrus deodara
root oil and its Pharmacokinetic and
Pharacodynamic studies, with reference to anti
ulcer and anti fungal effects. Ph.D. Thesis, Baqai
Medical University, Super Highway, Karachi,
Pakistan.
RAO, I.G. AND SINGH, D.K. (2001). Combination of
A. indica and C. deodara oil with PBO, MGK 264 an Embolia ribs against acuminata.
Chemosphere. 44 (8): 1691-95.
SANKARANARAYAN, R., KRISHAHAPA, S.,
BISARYA, S.C. AND DEV, S. (1973). Pryone
from Cedrus deodara essential oil. Tetrahedron
Letters (6): 427 1981 428.
SHIRWAIKAR, A., RAJENDRAN, K., KUMAR, C.D.
AND BODLA, R. (2004). Antidiabetic activity of
aqueous leaf extract of Annona squamosa in
streptozotocin-nicotinamide type 2 diabetic rats.
(online) Journal of Ethnopharmacology. Pp. 1-8.
STRELOKE, M., ASCHER, K.R.S., SCHMIDT, G.H.
AND NEUMANN (1989). Vapor pressure and
volatility of ß-asarone, the main ingredient of an
indigenous stored-product insecticide, Acorus
calamus oil. PHYTOPARASITICA 17(4): 299314.
TARIQ, R.M. AND NAQVI, S.N.H. (2006). Avian
toxicity of Acorus calamus (AC) and Annona
squamosa (AS) oil against chicken (poultry).
Pakistan j. entomol. Karachi 21 (1&2): 29-30.
TARIQ, R.M. AND QADRI, S.S. 2001. Repellent
activity of some local plant’s oil, two commercial
repellents, di-methyl phthalate and non-alcoholic
itter against dengue vector mosquitoes. Pakistan
j. entomol. Karachi, 16 (1&2): 7-10.
TARIQ, R.M., NAQVI, S.N.H., ZAFAR, S.M.N. AND
SAQIB, T.A. (2004). Piscean toxicity of rhizome
stem oil of Acorus calamus and seeds oil of
Annona squamosa against Labeo rohita and
Cyprinus carpio. Pakistan j. entomol. Karachi, 19
(1 & 2): 33-34.
TISSER, R AND ROBERT, M.C. (1995). Essential oil
safety. A guide for Health Care Professional.
Churchill Livingstone Publications. USA (ISBN
0443052603).
US ENVIRONMENTAL PROTECTION AGENCY
(1993). Azadirachtin tolerance Exemption,
Federal Register Vol. No. 30 Rules and
Regulations.
Pakistan j. entomol. Karachi 23 (1&2): 5-10, 2008
TAXONOMIC STUDIES OF SERGENTOMYIA FREETOWNENSIS SINTON
1930, SINDHICUS n. sub sp. (DIPTERA: PSYCHODIDAE)
IN SINDH, PAKISTAN
JUMA KHAN KAKARSULEMANKHEL
Investigator of Sand flies, Leishmaniases, Helminths, Mosquitoes & Ticks, Department of Zoology,
University of Balochistan, Saryab Road, Quetta, Pakistan
[email protected], Cell. # 0333-7860240
ABSTRACT
In the survey, the work was done to develop taxonomic record of sand fly Sergentomyia (Parrotomyia)
freetownensis Sinton, 1930, sindhicus n. sub sp. collected for the first time from new epidemic localities of
cutaneous leishmaniasis in Sindh Province (Pakistan). Since the specimens of the new sub species were
collected first time from the Sindh Province, therefore, it is named sindhicus due to its collection site. In view
of the published reports about the detection of encephalitis viruses from the species of the genus
Sergentomyia Franca and Theodor as well as from genus Phlebotomus Rondani and Berte, from the Indian
localities and European countries, the correct identification of the sand fly species becomes of significant
value in the study of epidemiology of leishmaniases and other viral diseases. The new sub species
sindhicus differs from the species S. freetownensis Sinton, 1930 in number of cibarial teeth, presence of 1
dozen punctiform fore teeth at the base of parallel teeth and ratio of anterior part of pharynx to its posterior
part. Therefore, in order to facilitate Zoologists and Medical researchers in correct identification, taxonomic
characters of S. freetownensis Sinton, sindhicus n. sub sp. were studied in detail with special reference to
its mouth parts, male and female genitalia and findings are presented in the present paper. Key to the
already known sub species of S. freetownensis Sinton is also formulated in order to distinguish the new sub
species.
Key words: Sand flies, Sergentomyia freetownensis Sinton 1930, sindhicus n. sub sp., Sindh.
INTRODUCTION
Phlebotomine sand flies are the proven vectors
of leishmaniases, sand fly fever and viral diseases.
There are about 700 species of sand flies of which
about 70 are considered to transmit diseases to
people.
Pakistan has several endemic foci of
leishmaniases and the disease is spreading
continuously (Rowland et al., 1999; Kolaczinski
et al., 2004; Kakarsulemankhel, 2004; Hashiguchi
et al., 2005; Khan, 2005; Kolachi et al., 2005; Wakil,
et al., 2006., Ahmad et al., 2008., Bari and Rahman,
2008).
Previous studies of the sand fly fauna of
Pakistan have been fragmentary. No comprehensive
taxonomic work exists in facilitating the identification
of Pakistani sand fly species except a few localized
surveys (Nasir,1958; Barnett and McDonald,1964;
Aslamkhan and Barnett, 1966,1967; Aslamkhan and
Rafiq, 1980; Burney and Lari 1986; Rab et al. 1986;
Safi, 1993; Aslamkhan 1996; Aslamkhan et al.
1997,1998). They have mentioned only the names of
the species they collected. Lewis (1967) while
studying west Pakistani sand flies, could not record
Sergentomyia freetownensis Sinton from Pakistan.
Species of genus Sergentomyia Franca and Parrot
are prevalent in the country. There are published
reports of transmission of encephalitis viruses
(Toscana virus-TOSV-Family Bunyaviridae, genus
Phlebovirus) from the species i. e. Phlebotomus
pernicious Newstead and Phlebotomus perfiliewi
Parrot of the genus Phlebotomus Rondani & Berte
from Italy (Verani
et. al.,1988), in
Spain
(Sanbonmatsu-Gamez et al., 2005), in southern
France in Sergentomyia minuta (Rondani) (Charrel
et al., 2006), Sabova virus in Phlebotomus of
Senegal (Bay et al.,1999), and in India also
(Geevarghese et al., 2005). In the light of these
findings, the correct identification of the species of
sand fly becomes of significant value in the study of
epidemiology of leishmaniases and other viral
diseases. In addition, the modern interest in
zoonoses, animal reservoirs of leishmaniases and
the role of sand flies as vectors, all have greatly
been focused and the significance of the correct
identification of sand flies is highlighted. During the
routine entomological collection, specimens of S.
freetownensis Sinton sindhicus n. sub sp. were
captured by the present author from human
residences of new endemic places of leishmaniases
(Jacobabad and Dadu) in Sindh Province.
Taxonomic characters of this fly especially mouth
parts and genitalia were studied and results are
given in the present paper. Key is also formulated of
subspecies of S. freetownensis Sinton and
relationships with its closest allies are also briefly
discussed. This attempt hopefully, will not only
serve as base line study of the taxonomy of
Pakistani sand flies but will also guide researchers in
correct identification of these flies.
The present investigation was carried out on the
materials (18 specimens of S. freetownensis Sinton
sindhicus n. sub. sp. collected from the field, human
residences, colleges, schools and hostels by means
Juma Khan Kakarsulemankhel
6
of sucking tubes and sticky papers in day and night
times in Sindh Province (Jacobabad and Dadu)
during May, 2006. Collected flies were transferred in
vials containing 70% alcohol and shifted to Zoology
Department, University of Balochistan, Quetta,
where descriptive observational study was carried
out. Each fly was processed and dissected following
the conventional techniques (Young and Duncan,
1994). Characteristics structures were studied,
measured, taxonomic notes were prepared and their
photographs were taken through camera mounted
Olympus microscope (BX41). Most structures were
measured with magnification of X100. All given
measurements are in mm. The data of specimens
critically examined for the description and
measurements are designated under “Material
examined”. Measured taxonomic characters are
those suggested by Dedet et al. (1991). Identification
of specimens was carried out with the help of
available literature (Kirk and Lewis, 1951; Lewis,
1967,1978). Prepared permanent slides were
deposited with the author’s collection of sandflies,
Department of Zoology, University of Balochistan,
Quetta.
pharynx with posterior width twice anterior width,
strongly constricted posteriorly, armature fairly
long,
denticles
stronger
and
more
numerous…freetownensis var. niger Parrot and
Schwetz (Belgian Congo, Nigeria, Ivory Coast,
Uganda).
-
3. ♀ pharyngeal teeth much smaller, some times
appearing as rows of points………freetownensis
var. sudanicus Theodor (Sudan and Kenya)
-
48-50 parallel narrow buccal teeth arranged on
an arc slightly concave posteriorly, with 10-12
anterior slightly pointed denticles………….........4
4.
♀ pigmented area brown, covering the whole width of buccal
cavity and prolonged anteriorly by a clear triangular
extension, pharynx a little less than twice as wide posteriorly
as
anteriorly
with
numerous
short
pointed
teeth……….…...freetownensis var. longoir Parrot (Abyssinia)
-
60-65 parallel, straight buccal teeth, at the bases
of these teeth a row of about a dozen punctiform
denticles……………………………………………5
RESULTS
S. freetownensis Sinton is known from one
female, from Sierra Leone. However, in the work of
Kirk and Lewis (1951) the name of africanus was
regarded as synonym of P. (S.) squamipleuris
Newstead and P. (S.) freetownensis Sinton becomes
the typical example of the various forms previously
known as P. africanus Newstead and its varieties.
Sinton (1930) discussed basic characters of S.
freetownensis. Buccal cavity has about 60 narrow
parallel teeth on an arc concave posteriorly, flask
shaped pharynx, about 2.5 times as wide posteriorly
as anteriorly, armed with numerous backwardly
directed armature, spermatheca an oval capsule
twice as long as wide, ending in a narrow duct.
Key to the sub species of S. freetownensis Sinton
1. ♀ antennal formula 2/3-15, A3= 0.135-0.17,
cibarial teeth 40-50 parallel arranged on a
concave arc, pigmented plate sausage shaped,
concave posteriorly and with a paler, triangular,
pointed, forward extension, pigmented plate
triradiate with thickened lateral arms, pharyngeal
spines backwardly directed, spermatheca an
elongated capsule opening into a relatively
narrow duct…freetownensis var. magnus Sinton
(Belgian Congo and Transvaal).
-
60-70 uniform parallel buccal teeths on an arc
slightly concave, no anterior denticles….………2
2. ♀ pigmented plate black, and sausage shaped,
occupying nearly the whole width of the cavity,
with an anterior sub triangular pale prolongation,
30-33 buccal teeth, and a small point like
denticle at the base of every second tooth in the
armature……………………………………………3
5. ♀ dark brown pigmented plate in the form of
crescent slightly concave possteriorly, not
occupying whole width of buccal cavity, a
paler anterior triangular prolongation, pharynx
with
well
developed
pointed
spines
freetownensis var. eremitis Parrot and de
Joliniere (Sudan).
-
46- 48 straight long, parallel buccal teeth on an
arc…………………………………………………..6
6. ♀ pigmented plate black, not occupying the
whole width of buccal cavity and with a small
triangular
anterior
prolongation,
pharynx
constricted posteriorly, armed with moderately
long denticles……….……freetownensis var. ater
Parrot (Abyssinia)
-
20 large, pointed buccal teeth with their bases
nearly in straight line, about three median and
one or two of the extreme lateral teeth shorter
than the rest, a row of fine punctiform teeth……7
7. ♀ pigmented plate large, mushroom shaped,
pharynx armed with numerous sharply pointed
teeth posteriorly.......................freetownensis var.
meridianus
De
Meillo
and
Lavoipierre
(Transvaal, South Africa)
Sergentomyia freetownensis Sinton, sindhicus
n. sub. sp. (Figures 1A–1G)
Phlebotomus minutus var africanus, Newstead, 1912,
Bull. ent. Res., 3: 363; Adler & Theodor, 1926, Bull.
ent. Res., 16: 401, Ann trop. Med. Parasit., 21: 63.
P. africanus, Adler et al. 1929, Rev. Zool. Bot. Afric. 18:72.
P. fensis Sinton, 1930, Ind. J. med. Res., 18: 171-193.
Taxonomic studies of Sergentomyia freetownensis Sinton 1930, Sindhicus n. sub sp.
Wings (X100)
Distribution
Narrow (1.50 long, 0.35 broad) and pointed,
α/β=0.66, δ=0.05, П=0.10, gamma= 0.25.
Pakistan: Sindh: Jacobabad and Dadu.
7
Comparative note
Palps and antennal segments (X100)
Palps 0.53 long, ratio 1, 2.6, 3.3,4, 7, formula 1, 2, 3,
4, .5. 3rd antennal segment 0.13 long, segments 4th
and 5th each 0.07 long. Position of ascoid on A3
0.76, and at A4 and A5 each 0.28 of the segments.
Mouth parts, Cibarium and Pharynx (X100)
(Figs.1A–F) Proboscis 0.20 long. Labrum 0.16 long,
strong chitinous structure, sides parallel, apex bluntly
pointed, and margins furnished by a series of long
very short leaf like sensillae closely together and
numbering about eight on either side. Mandible 0.16
long, blade like structure, outer edges markedly
serrated with sharp but minute teeth (Fig. 1A).
Dental depth of 0.05. Maxilla 0.15 long, basally stout
but narrows very much towards its apex (Fig.1B),
much narrower than mandibles, one edge is pointed
with about five relatively larger dot like chitinous
lateral teeth, widely separated, the opposite edge
with smaller ones set closely together (Fig.1C) and
numbering about 34 ventral teeth. Dental depth 0.09.
Hypopharynx 0.16 long, marginal leaf like sensillae
very much shorter, apical part is broadly concave
and in its center a salivary duct runs. Cibarium 0.05
broad, comb-like cibarial armature (Fig.1D)
consisting of about 65 parallel, straight teeth, the
extreme lateral ones are slightly bigger in length
than the median ones, teeth standing on a slightly
curved arch, anterior to it, there is a row of about
dozen punctiform denticles, pigment patch 0.04 long
and 0.01 broad (Fig.1E), brown in color with a
yellowish short triangular extension, pigmented area
not occupying the whole of the cibarium. Pharynx
0.13 long, flask shaped, slightly widened at its
posterior half but narrowing again towards base,
posterior breadth is 1.66 of the anterior breadth. The
anterior edge of armature forms an almost straight
line, most of the armature confined to the basal part
of the pharynx (Fig.1F).
Female genitalia (X100)
Spermatheca (Fig.1G) cylindrical, barrel-shaped,
0.08 long, 0.03 broad, narrow duct leaves from each
spermatheca and at some distance unite with one
another to form a joint single duct finally open in to
genital atrium.
♂ not known from Sindh during present survey.
Material examined
8 ♀, Sindh, May, 2006.
This new sub species shares some of its characters
such as comb-like buccal armature, flask shaped
pharynx, constricted posteriorly and oblong
spermatheca with magnus, ater, eremitis, longior,
niger, sudanicus, meridianus. It is distinct however,
in its group in having brown colored pigment patch,
about 65 parallel straight buccal teeth, with extreme
lateral ones slightly bigger in length than the median
ones, and with a row of about one dozen punctiform
denticles anterior to larger teeth, pharynx 1.6 X as
wide posteriorly as anteriorly.
DISCUSSION
According to Lewis (1978) S. africana complex is
mainly African and includes a number of forms which
have a comb-like cibarial armature and oblong
spermatheca and have been variously treated as
several species or as one by Quate (1964). Lewis
(1974) pointed out that due to some degree of intergradation between some named forms, various local
variants have come into existence. Therefore, Lewis
(1978) treated it as S. africana magna (Sinton) form
asiatica Theodor stat. n. The cibarium of ♀ has 4550 teeth in a concave row and no central patch of
fore teeth, no notch in ventral plate, pigment patch
concave posteriorly and has a simple forward
process, pharynx with long teeth and spermatheca is
oblong (Lewis (1967, 1978) and for these reasons,
Lewis (1974) regarded S. a. magna as a sub
species. However, ♀ S. freetownensis Sinton have a
triradiate pigmented area with thickened lateral
arms, cibarial teeth about 60, arranged on a concave
line, pharynx flask shaped, the broadest posterior
part being 2.50 times as broad as its anterior part
(Sinton, 1930).Since Pakistani specimens was not
available even Lewis (1978) suggested that
Pakistani specimens collected from Kandhkot and
Shikarpur by Lewis (1967) probably belongs to a
small northern form found in Morocco, Israel and
India, which Abonnenc and Yvore (1969) and
Abonnenc (1972) known as S. africanus Newstead
and its varieties.
Kirk and Lewis (1951) while discussing Ethiopian
sand flies, they suggested that P. africanus
Newstead (Adler et al. 1929) from Belgian Congo is
a synonym of P. squamipleuris Newstead 1912, and
P. freetownensis Sinton, 1930 becomes the typical
example of various forms. Two species of africanus
group i. e. S. a. magna Sinton form asiatica Theodor
stat. n. which was reported and described from
Sindh (Kandhkot and Shikarpur) by Lewis (1967)
8
having ♀ cibarium 45-50 parallel teeth in a concave
row no punctigorm fore teeth, and oblong
spermatheca and S. africana Newstead reported
from Uthal, Balochistan by Rab et al.(1986), details
of figures and diagnostic characters were not
supplied. The present new sub species i. e. S.
freetownensis Sinton sindhicus, n. sub sp. is quite
differentiated from the above mentioned sub species
in characteristics like more number (65) of cibarial
teeth, parallel, and straight, extreme lateral ones
slightly larger in size than the middle ones, and with
a row of about 12 punctiform denticles at the base of
larger buccal teeth, pharynx 1.6 time as wide
posteriorly as anteriorly, pigmented area not
occupying the whole of the cibarium. Therefore, in
the light of these characters, present specimens of
Sindh fully deserve to be treated as a new sub
species. Its name sindhicus is given due to its
locality (Sindh). Morph metric measurements and
morphology
of
labrum,
mandible,
mxilla,
hypopharynx, and cibarium of the of the new sub
species can not be compared with the other sub
species as the same has not been furnished by the
earlier workers like Sinton (1930), Lewis (1967,
1974, 1978) and Rab et al. (1986).
However, the new sub species has a very
localized and restricted distribution in Sindh and
specially its presence in the human residences of the
endemic foci of leishmaniases and its non
established vectorial capacity clearly demands to
initiate a comprehensive program for the control of
sand flies and leishmaniases. It is hoped that the
present findings would provide a basis for the further
research on sand flies taxonomy.
9
REFERENCES
Fig.
Sergentomyia
(Parrotomyia)
freetownensis
Sinton sindhicus n. sub sp. A, Mandible; B, Maxilla
showing lateral teeth; C, Maxilla showing ventral
teeth; D, Cibarium showing cibarial teeth; E, Pigment
patch; F, Pharynx showing pharyngeal armature at
its base; G, Spermatheca.
ABONNENC, C. (1972). Les phlebotomes de la
region Ethiopienne (Diptera, Psychodidae).
Mem. Off. Rech. sci. tech. Outre-mer no. 55:1289.
ABONNENC, C. AND YVORE, P. (1969).
Phlebotomes du complex africanus (Diptera,
Psychodidae). Cah. Off. Rech. sci. tech. Outremer. Ent. med. 7: 181-208.
ADLER, S., THEODOR, O. AND PARROT L. (1929).
Phlebotomes du Congo Belg. Rev. Zool. Bot.
Afric. 18: 72-89.
AHMAD, I., HUMAYUN, Z. AND AHMED, M. (2008).
Patterns of cutaneous Leishmaniasis cases
among troops and their families in Sibi. Pak.
Armed Forces ed. J. 58: 209-212.
ASLAMKHAN, K. (1996). Biodiversity of sand flies
hlebotominae) of Pakistan. M. Sc. Thesis.
Department of Zoology, Govt. College, Lahore,
Punjab Univ. Pakistan, 144 pp.
ASLAMKHAN,
K.,
ASLAMKHAN,
M.
AND
AZIZULLAH. (1997). The distributional records
of sand flies (Phlebotominae) of Pakistan and
Kashmir from 1908 to 1996. Pak. J. Zool. 29:
351-360.
ASLAMKHAN,
K.,
ASLAMKHAN,
M.
AND
AZIZULLAH (1998). Biodiversity of sand flies of
Pakistan and Kashmir. Pak. J. Zool. 30: 13-21.
ASLAMKHAN, M. AND BARNETT, H. C. (1966).
Studies on the bionomics of sandflies. Ann.
Rept. Uni. Md. Sch. Med. ICMR. 54-65.
10
ASLAMKHAN, M. AND BARNETT, H. C. (1967).
Studies on sand fly fever and on the bionomics
of sandflies. Ann. Rept. Uni. Md. Sch. Med.
ICMR. 129-157.
ASLAMKHAN, M. AND RAFIQ, S. (1980). Studies
on cutaneous leishmaniasis and sand flies of
Balochistan. Ann. Rept. Uni. Md. Sch. Med.
ICMR. 315-324.
BARI, A.U. AND RAHMAN, B. (2008). Many faces of
cutaneous leishmaniasis. Ind. J. Dermatol.
Venerol. Leprol. 74: 23-27.
BARNETT, H. C., AND MCDONALD, W. A. (1964).
Studies on sand fly and sandfly fever. Ann.
Rept. Uni. Md. Sch. Med. ICMR. 54-56.
BA. Y., TROUILLER, J., THONNON, J. AND
FONTENILLE, D. (1999). Phlebotomus of
Senegal: Survey of the fauna in the region of
Kedougou. Isolation of arbovirus. Bull. Soc.
Patho. Exotiq. 92: 131-135.
BURNEY, M. I. AND LARI, F. A. (1986). Status of
cutaneous leishmaniasis in Pakistan. Pak. J.
Med. Res. 25: 101-108.
CHARREL, R. N. A., IZRI, S., TEMMAM, X., DELAMBALLERIE, AND PAROLA, P. (2006).
TOSCANA-virus in Sergentomyia minuta flies.
Letter to the Editor. Emerg. Infect. Dis. 12: page
1-2.
DEDET, J. P., GOMES, E. L., FERREIRA-RANGEL,
R., FALCAO, A. L., GALATI, E. A.B.,
BERMUDEZ, H., HERRERO, M. V., FERRO. C.,
FELICIANGELI, D. AND HERVAS, D. (1991).
Proposition of a standard description for
Phlebotomine sand flies. Parassitologia, 33
(Suppl.): 127-135.
GEEVARGHESE, G., ARNAKALLE, V. A., JADI, R.,
KANOJIA, P. C., JOSHI, M. V. AND MISHRA,
A. C. (2005). Detection of Chandipura virus from
sandflies in the genus Sergetomyia (Diptera,
Psychodidae) at Karim Nagar District, Andhra
Pradesh, India. J. Med. Entomol. 10: 495-496.
HASHIGUCHI, Y., BARROSO, P. A., MARCO, D. J.,
BHUTTO,
A.
M.,
SOOMRO,
F.R.,
KAKARSULEM-ANKHEL, J. K., et al., (2005).
Cutaneous
eishmaniasis
in
Sindh
and
Balochistan, Pakistan, with special reference to
vector sand flies and causative agents. Proc. 5th.
Int. Symp. on Phlebotomine sand flies. Arch. de
L’ Inst. Pasteur de Tunis Jan-Jun, 82: 81.
KAKARSULEMANKHEL, J. K. (2004). Present
situation of cutaneous leishmaniasis n
Balochistan, Pakistan. Pak. J. Bio. Sci. 7: 698702.
KHAN, Z. (2005). Cutaneous leishmaniasis in N.
W.F.P. J. Postgrad. Med. Inst.19:226-228.
KIRK, R. AND LEWIS, D. J. (1951). The
Phlebotominae of the Ethiopian region. Trans. R.
Ent. Soc. Lond. 102: 383-510.
KOLACHI, H. B., DAHAR, M. Y., RATHI, S. L., AND
KHASKHELI, A. (2005). Epidemic of cutaneous
leishmaniasis in Taluka Johi, District Dadu,
Sindh. Infect. Dis. J. Pak. Apr-Jun: 37-40.
KOLACZINSKI, J., BROOKER, S., REYBURN, H.
AND ROWLAND, M. (2004). Epidemiology of
anthroponotic cutaneous leishmaniasis in
Afghan refugee camps in north-west Pakistan.
Trans. R. Soc. Trop. Med. Hyg. 98: 373-378.
LEWIS, D. J. (1967). The phlebotomine sand flies of
west Pakistan (Diptera, Psychodidae). Bull. Brit.
Mus. Nat. Hist. (Ent.), 19: 1-57.
LEWIS, D. J. (1974). The biology of the
Phlebotomidae in relation to leishmaniasis. Ann.
Rev. Ent. 19: 363-384.
LEWIS, D. J. (1978). The phlebotomine sand flies
(Diptera: Psychodidae) of the Oriental Region.
Bull. Brit. Mus. Nat. Hist. (Ent.), 37: 217-343.
NASIR, A. S. (1958). Sandfly fauna in west Pakistan.
Pak. J. Hlth. 8: 21-22.
QUATE, L. W. (1964). Phlebotomus sand flies of the
Paloich area in the Sudan (Diptera, Psychodiae).
J. Med. Entomol. 1: 213-268.
RAB, M.A., AZMI, F.H., IQBAL, J., HAMID, J.,
GHAFOOR, A., BURNEY, M.I. AND RASHTI,
M.A. S. (1986). Cutaneous leishmaniasis in
Balochistan: Reservoir hosts and sand fly
vectors in Uthal-Lasbella. J. Pak. Med. Assoc.
36: 134-138.
ROWLAND, M., MUNIR, A., DURRANI, N. NOYES,
H. AND REYBURN, H. (1999). An out break of
cutaneous leishmaniasis in settlements of
Afghan Refugees in North West Pakistan. Trans.
R. Soc. Trop. Med. Hyg. 93: 133-136.
SAFI, F.S. (1993). A contribution to the sand fly
fauna of Peshawar. M.Sc. Thesis. Government
College Lahore, University of Punjab, Pakistan,
61 pp.
SANBONMATSU-GAMEZ, S., PEREZ-RUIZ, M.,
COLLAO, X., SANCHEZ, F., ROSA-FRAILE, M.
DE-LA. et al. (2005). Toscana virus in Spain.
Emerg. Infect. Dis.11: 1701-1707.
SINTON, J. A. (1930). Some new species and
records of Phlebotomus from Africa. Ind. J. Med.
Res. 18: 171-193.
VERANI, P., CIUFOLINI, M. G., CACIOLLI, S.,
RENZI, A., NICOLLETTI, L., SABATINELLI, G.
et al., (1988). Ecology of viruses isolated from
sand flies in Italy and characterization of a new
Phlebovirus (Arbo-virus). Am. J. Trop. Med. Hyg.
38: 433-439.
WAKIL, A., BILQEES, F. M. AND SALIM, (2006).
Cutaneous leishmaniasis in Dadu district during
2001-2004. Proc. Parasitol. 41: 19-39.
YOUNG, D. G. AND DUNCAN, M. A. (1994). Guide
to the Identification and Geographic distribution
of Lutzomyia sand flies in Mexico, the West
Indies, Central and South America (Diptera:
Psychodidae). Mem. Am. Entomol. Inst. 57: 1881.
COMPARATIVE TOXICITY OF ORGANIC SOLVENTS AND
INSECTICIDESAGAINST CABBAGE BUTTERFLY
SEEMA TAHIR1, TAHIR ANWAR1, MUHMMED SAMIULLAH CHANNA1, IMTIAZ AHMED2
AND YOUSAF HAYAT KHAN3
1. Pesticide Research Institute, Southern-Zone Agricultural Research
Pakistan Agricultural Research Council, University Campus, Karachi-75270, Pakistan.
2. Department of Zoology, University of Karachi, Karachi-75270, Pakistan
3. Ecotoxicology Research Institute, National Agricultural Research Centre, Islamabad.
ABSTRACT
Under dose response observation chloroform was found to be more toxic with 60% mortality after 24 hours
followed by the methanol (56.6%), benzene (53.3%) and acetone (40%) when applied topically on the 3rd
instar larvae of cabbage butterfly. DDT was found to be toxic when tested against CBF caterpillar with other
pesticides like ϒ-BHC and chlorpyriphos.
KEYWORDS: Insecticides, solvents, toxicity.
INTRODUCTION
Large cabbage butterfly, P. brassicae L. is
one of the greatest pests of cruciferous plants
in the peak periods of its abundance, causing
serious change on cabbage cultivation. Their
larvae attach the leaves stems; inflorescence
pots of the host plant and lower the yield of
the seed. These are in delay distribute
throughout the Asia and Europe. In Pakistan it
is reported as a major pest of cauliflower
(Ghouri 1960) and causes heavy losses to
cabbage and cauliflower (Mushtaq and
Mohuddin, 1984).
Atwal (1976) recommended Malathion
(0.05%) Diazinon (0.02%) or carbaryl (0.02%)
for the control of this pest. In Pakistan the
chemical control of this pest has been
reported by many workers. (Gupta et al. 1985;
Anwar et al. 1989 & 1995). The sub lethal,
anti-feedant
&
repellants
effects
of
insecticides have been studied by Blackwell
(1988) & Ten (1981). Insecticides used to
maintain the population at less than damaging
levels when received little attention as to their
role in a management scouting program
utilizing the threshold concept. Growers
presently used many different insecticides to
control lepidpoterous pest of cabbage.
The objective of our study to evaluate the
efficacy of commonly used and nonregistered material and banned chemicals
(usually smuggled from neighbour countries
and used in remote areas of Sindh and
Punjab). With toxicity of solvents also. The
present investigations showed that DDT and
Chlorpyriphos were equally toxic when
applied topically on the thoracic region of 3rd
instars larvae of cabbage butterfly (CBF) after
24 hours of treatment.
The larvae of cabbage butterfly (CBF)
were collected from the cabbage field from
Sialkot. Larvae were brought to Entomological
Research Laboratory, National Agriculture
Research Centre, Islamabad. The larvae
were fed on fresh cauliflower leaves. Different
larval stages of cabbage butterfly were held in
cage of 45 x 45 x 45cm made up of wooden
frame from sided were provided with wire
screen of 20 mesh and bottom were made up
of ply wood. One side of cage had on opening
provided with sieve of mouslin cloth for
cleaning and feeding purpose, temperature
and relative humidity were maintain at 25oC
and 50% respectively.
Different concentrations 0.01, 0.025, 0.05
and 0.1% of DDT and Chlorpyriphos were
prepared using acetone as solvent. Three
replicates with batches of 10 insects were
used for each treatment. A batch of control
(no treatment) and check (acetone) were also
kept. Treated insects were keep in petridishes
(90mm) and provided fresh leaves of cabbage
with control and check. They were placed in
rearing room at temperature of 25oC and 50%
humidity. Observations for mortality were
Tahir et al.
12
taken (14.1 LD) at 6 hrs, 12 hrs, 24 hrs and
48 hrs after treatment. DDT was tested
against 2nd instar larvae (0.2g/10 larvae). 1µl
of the test concentration was topically applied
on dorsal side of thoracic region with the help
of
hand-mounted
micro-applicator.
Chlorpyriphos was tested against 3rd instar
larvae (2.0g/10 larvae). 0.1µl of the test
concentration ranging from 0.01 to 0.1% was
injected on lateral side of thoracic region with
the help of electrical micro-applicator (Bukard,
Manufacturing Co. Ltd., Reckmons Worth,
England. Variations in number of mortality for
treatment were analyzed by means of
Analysis of Variance; more were separate
(P=0.05) by Ducans (1955) multiply range
test Probit analysis of C50 and LC90 after 24
hours and 48 hours of the treatment.
RESULTS AND DISCUSSION
The present investigation showed that
acetone, methanol and chloroform and
benzene which are known to be used as
solvents also possess toxic properties at
different concentration when applied topically
on the thoracic region of 3rd instar larvae of
Pieres brassicae. Chloroform found to be
more toxic with 60% mortality after 24 hours
followed by the methanol (56.6%), benzene
(53.3%) and acetone (40%) while mortality
evaluation against other pest species has
shown acetone to be more toxic than
methanol. The results are given in Table-1 &
Fig.1.
Topical bioassay application has been
used successfully on different test species
evaluation of toxicity of different insecticides
by topical application of the larvae of P.
brassicae has been reported (Choudhary et
al., 1988, Durha and Hameed, 1988, Kasana
et al., 1996, Tahir et al., 1999).
Endosulfan was found to be more toxic
than other tested insecticides when applied
topically CBF larvae (Mahabin et al., 1992)
which is in favor with the present result that
topical application of DDT and Chlorpyriphos
was found to be less toxic.
Sharma and Nath (1992) evaluated the
bio efficacy of insecticide against CBF larvae
and observed that toxicity of fenvalerate,
HCH, methyl parathion, and DDT is almost
same. Similarly in the present study the toxic
effects of both insecticides against 3rd instar
larvae of CBF were found.
Sharma and Nath (1992) observed that
the larvae of CBF were found to be more
susceptible to DDT when applied topically
which is fully compatible with present findings.
Anwar et al. (1995) observed the maximum
larvae mortality i.e. 87.50% after 24 hours
and 92.5% after 48 hours of treatment when
Dimlore (Chlorpyriphos and Dimethoate) the
applied on CBF larvae as chlorpyriphos is
also found to be more toxic to CBF in the
present study.
Singh and Jain (1987) observed the
toxicity of various solvents against housefly
when
screening
plant
products
as
insecticides. It was found that acetone,
methanol, benzene and chloroform when
applied topically on thoracic region were
found more toxic, while it was noted that as
the carbon chain of solvent increased the
toxicity also increased as tested Hydrocarbon
solvent on the toxicity of Cypermethrin to 3rd
instar larvae of S. Litura (Jaglan and Sircar
2003).
In past the DDT was commonly used
against the larvae of P. barssicae. Ripper et
al. (1948) observed the toxicity of DDT
against P. barssicae larvae when applied
DDT particles with substance digestible only
by specific insects which may be useful in
acquiring the destruction of beneficial insects
at the same time penetration of DDT through
cabbage butterfly eggs killed the larvae first
after hatching (Beament & Lal, 1957).
Similarly the DDT was found to be toxic
when tested against CBF caterpillar with other
pesticides like ϒ-BHC diazinon, dichlorvos &
carbaryl (Verma et al., 1969). These pervious
findings are in full agreement with present
results that relative toxicity of DDT is also like
other pesticides i.e. Chlorpyriphos against
CBF.
In the lab experiments it was found that
acetone and acrolein vapors could penetrate
into the wheat mass and kill concealed
insects in interkernel spaces while they were
harmless to wheat seed viability (Poumirza &
Tajbakhsh 2002 & 2008).
While in present study acetone was found to be
toxic when applied topically on CBF. Similarly the
acetone toxicity LC50 was found to be 15.40, 15.51,
17.55 & 18.26 µl/L respectively against Oryzaephilus
surinamensis (L), Callosobruchus maculates (F),
Tribolium confusum (Duv.) and Sitophilus granaries
(L.) respectively.
Comparative toxicity of organic solvents and insecticides
13
….
Fig. 1. Time course study of DDT and Chlorpyrifos (CHP) applied topically at the rate of 0.1µl
on the thoracic region against 3rd instar larvae of cabbage butterfly, Pieris brassicecae.
Table 1. Lethal Doses of DDT and chlorpyrifos at 95% confidence level after 24 and 48 hours
of treatment against larvae of P. barssicae (cabbage butterfly).
S. No.
Insecticides
1.
DDT
2.
3.
CHF
4.
Time
(hrs)
24
LD50s
Range
Regression line (non weighted)
0.36
0.022-0.058
Y=4.945 + 1.416 (X-8.524)
48
0.020
0.011-0.028
Y=5.394 + 1.828 (X-8.524)
24
0.040
0.021-0.067
Y=4.781 + 1.954 (X-8.524)
48
0.016
0.000-0.035
Y=5.287 + 0.958 (X-8524)
LD90s
5.
DDT
6.
7.
8.
CHF
24
0.279
0.130-2.417
Y=4.945 + 1.416 (X-8.524)
48
0.099
0.062-0.269
Y=5.394 + 1.828 (X-8.524)
24
0.222
0.109-3.060
Y=4.781 + 1.954 (X-524)
48
0.358
0.104-0.245
Y=5.287 + 0.958 (X-8.524)
Tahir et al.
14
REFERENCES
Indian Journal of Plant Protection, 20: 2, 202204.
ANWAR, T., S. TAHIR, A. JABBAR AND A.A.
HASHMI (1995). Field evaluation of novel
pesticides against cabbage butterfly, Pieris
barssicae (L.) (Pieridae: Lepidoptera).
MUHAMMAD ZAMAN (1986). Effect of four
pyrethroids on the insect pests of rape. Pakistan
J. Sci. Ind. Res., Vol. 29, No.3.
ANWAR, T., A. JABBAR, A. MAJID, S. TAHIR AND
S.N.H. NAQVI (1989). Antifeedant activity of
juliflorine against larvae of brassicae (L.). J. Pak.
Entomol. Kararachi, 41(1-2): 25-32.
MUHAMMAD ZAMAN (1989). Evaluation of some
insecticides against the mealy cabbage aphid
and cabbage butterfly on rape in Swat. Sarhad
J. Agric., Vol.5, No.4.
ATWAL, A.S. (1976). Agricultural pest of India South
East Asia. Kalyani Publishers, Delhi, India 479
p9.
MUSHTAQUE, M. AND MOHYUDDIN, A.I. (1984).
Pieris barssicae (Piendae Lepidoptera), a pest of
crucifers and its control by parasites. Pak. J.
Agric. Res. 5(3): 165-169.
BEAMENT, J.W.L.; AND LAL, R. (1957). Penetration
through the egg shell of Pieris barssicae. Bulletin
of Entomol. Res. 48 109-25.
BLACKWELL, A. (1988). A several and development
of Pieris barssicae larvae upon application of
sublethal doses
of chlordimeform.
Entomologia Experimentalis at Applicata, 48(2):
149-156.
CHOUDHARY, S.K.; SINGH, A.K.; SINGH, S.P.;
(1988). Evaluation of toxicity of some newer
insecticides against the larvae of cabbage
butterfly, Pieris barssicae L. Journal of
Research, 6:, 1-2, 88-90.
DUHRA, M.S.; HAMEED, S.F.; (1988). Toxicity of
some insecticides to the cabbage butterfly on
cauliflower. Journal of Research, 6: 1-2, 17-20.
DUNI CHAND SHARMA; MAHABIR SING; AMIT
NATH; SHARMA, D.C.; SINGH, M; NATH, A;
(1993).
Synergistic
properties
of
hexachlorocyclohexane. Indian Journal of
Entomology, 55: 4, 368-374.
GHOURI, A.S.K. (1960). Insect pest of Pakistan.
FAOPL. Prot Comm. S. Asia Pacific. Reg Tech.
Doc. 8:31.
GUPATA, P.R., VERMA, A.K. MISHRA, R.C. (1985).
Field efficacy of some insecticide against
caterpillars and thrips on cauliflowers crop.
Vegetable Science 12(1): 49-54.
JAGLAN, R.S.; SIRCAR, P. (2003). Effect of carbonchain of solvents on the toxicity of Cypermethrin
emulsion formulation against larvae of
Spodoptera litura (Fab.). Annals of Biology
(Hisar, India), 19(1), 81-83.
KASANA, A; MATIN, MA; RAFI, M.A. (1996).
Efficacy of Malathion against larvae of Pieris
barssicae. Pakistan Journal of Zoology, 27: 4,
359-361.
MAHABIR SINGH; CHANDEL, R; SHARMA, D.C.;
SINGH, M.; (1992). Toxicity of insecticides to
different larval instars of Pieris barssicae L.
POURMIZA, A.A.; TAJBAKHSH, M. (2002). Effect of
acetone in controlling stored pest insect.
Majallah-I Ulum-I Kishavarzi va Manabi-I Tabi
Danishgah-I Sanati-I Isfahan, 6(3), 229-239.
POURMIRZA. A.A. AND TAJBAKHSH. M. (2008).
Studies on the Toxicity of Acetone and Carbon
Dioxide on Stored-Poduct Insects and Wheat
Seed. Pak. Jour. Biol. Sci. 11(7): 953-963.
RIPPER, W.E.; GREENSLADE, R.M.; HEALTH, J.;
BARKER, K. (1948). New formulation of DDT
with selective properties. Pest Control Ltd.,
Harston, Cambridge, UK. Nature (London,
United Kingdom).
SHARMA, D.C., NATH, A; (1992). Effect of
hexachlorocyuclohexane on the bio-efficacy of
insecticides. Pesticide Research Journal, 4: 2,
111-115.
SINGH, D.; JAIN, D.C. (1987). Relative toxicity of
various organic solvents generally used in
screening plant products for insetticidal activity
against the house fly Musca domestica L. Indian
Journal of Experimental Biology, 25(8), 569-70.
TAHIR, S., ANWAR, T., KHAN, M.R., AZIZ, S.,
ILYAS, M. AND RAFI, M.A. (1999). Toxicological
of Dimlor (A Mixture of Eimethoate and
Chlorpyrifos
against
corn
leaf
Aphid,
Rhopalosiphum maidis (Fitch) under different
Micro-Climatic conditions. Pakistan J. of
Biological Sciences, 2(2): 315-317.
TEN, K.H. (1981). Antifeeding effect of Cypermethrin
and permethrin at sublethal levels against Pieris
barssicae Larvae. Pesticide Science, 12(6): 619626.
VERMA, A.N.; SHARMA, P.D.; SARAMMA, P.U.
(1969). Relative toxicity of some contact
insecticides against cabbage caterpillar, Pieris
barssicae (Pieridae; Lepidoptera) Journal of
Research (Punjab Agricultural University). 6(1)
(Suppl.), 197-9.
Pakistan j. entomol. Karachi, 23 (1&2): 15-17, 2008
REDESCRIPTION OF CAYSTRUS PYGMEAUS LINNAVUORI
(HEMIPTERA: PENTATOMIDAE) AN ISOLATED SPECIES
FROM ETHIOPIAN REGION
MUHAMMED ZAHID1 AND IMTIAZ AHMED2
1) Department of Zoology, Federal Urdu University of Arts, Sciences and Technology,
Gulshan-e-Iqbal Campus, Karachi.
2) Department of Zoology, University of Karachi, Karachi.
ABSTRACT
The species belonging to the stink bug genus Caystrus Stål pygmeaus Linnavuori is redescribed here
with reference to its colouration, head, pronotum, scutelum, hemelytra and important measurements
specially male genitalia from Barkina faso (earlier name upper Volta) south of Senegal in the western
Africa is considered here entirely isolated in its subclade C. trivalis (Gerstaecker) group.
Key words: Redescription, Caystrus pygmeaus, Ethiopian region.
INTRODUCTION
Linnavouri (1972) described C. pygmaeus
Linnavuori with reference to some of the external
characters of head & pronotum including some
measurements and male genitalia including some of
the characters of pygophore and paramere. He also
keyed his new species rather close to another of his
new species parviceps but considered pygmaeus
very distinctive in the genus. He described his new
species from Poundou, upper Volta (present name
Barkina Faso), south of Senegal in the western
Africa), on the basis of one male specimen which
was designated as holotype. The present 2nd author
had an opportunity to examine this species in
American Museum of Natural History (AMNH), New
York, by the courtesy of Dr. Toby Schuh incharge
entomological collections. Presently this species is
redescribed with special reference to its characters
of prnotum, scutelum, colouration & male genitalia. It
is considered entirely isolated in its C. trivalis
(Gerstaecker) group and to date its female genitalia
is unknown. Its relationship is described here for the
first time in its trivalis group.
The holotype of this species was examined by
the 2nd author of the present paper during the visit of
American Museum of Natural History, New York
(AMNH) by the courtesy of Dr. Toby Schuh incharge
entomological collections of that museum. For the
study of male genitalia the entire specimen with
pygophore was plunged in the boiling water for five
minutes. The pygophore in the softened specimen
was extracted and placed in warm 10% KOH
solution for a few minutes following the technique of
Ahmad (1986) & Ahmad and McPherson (1990,
1998) and the male genitalia specially the pygophore
and paramere were dissected and illustrated. The
male genitalial capsule with paramere and aedeagus
was then glued with the specimen.
RESULTS
Caystrus pygmaeus Linnavuori
(Figs. 1-3)
Caystrus pygmaeus Linnavuori 1972: 400-402;
1974: 402; 1982: 76.
Coloration and general shape
Body generally yellowish brown with brown
punctures; eyes brownish black; ocelli pinkish;
membrane of hemelytra hyaline; ovate.
Head
Slightly broader than long, anteocular distance
longer than remainder of head; paraclypei narrowed,
slightly longer than clypeus but not enclosing the
later; antennae short with second segment much
shorter than third, antennal formula 1<2<3<4<5;
labium reaching hind coxae.
Thorax
Pronotum about 2x broader than its length,
anterior margin slightly broader than head width
across the eyes, anterior angles acute, humeral
angles subacutely produced, lateral margins sinuate,
posterolateral margin distinctly sinuate.; scutellum
with elongated subrounded apical lobe.
16
Zahid & Ahmed
17
Key to close allies of C. pygmaeus
Male genitalia
Pygophore (Fig. 2) some what quadrangular,
dorsomedian
surface
deeply
concave,
ventroposterior margin medially knobed, lateral lobe
prominent,
truncately
produced;
proctiger
rectangular; paramere (Fig. 3) somewhat F- shaped,
outer margin convex, apex of blade beak shaped,
inner lobe broad with hairs.
Material Examined
One male 19. X 1927; Olsufjev; American Museum
of Natural History (AMNH)
Comparative note
This species is most closely related to
C. deserticolus Linnavuori in having head shorter
and lateral margins of pronotum narrowly lamellate
but it can easily be separated from the same in
having shorter length (9.0 mm), antennae short and
incrassate and puncturing of pronotum very coarse
and dense.
1. Puncturing
of
pronotum
sparse
and
fine………………………………………………….2
-
Puncturing of pronotum very coarse and
dense……………………………………………….3
2. Head much reduced in length (0.73 X as broad
as
long),
scutellum
sharply
triangular…………………………………edentatus
-
Head not much reduced as above, scutellum not
as above……………………....…………parviceps
3. Body of medium to large size, antennae
incrassate…………………..…….….mawambinus
-
Body of small size, antennae not as
above…….………………………………………...4
4. Lateral margins of pronotum straight only
extreme margin upcurved……...……deserticolus
-
Lateral
margins
of
pronotum
distinctly
sinuate……………….……………….…pygmaeus
DISCUSSION
Linnavouri (1972) gave the illustrations of head,
pronotum, pygophore and paramere but he
considered it most closely related to C. parviceps
although he considered it distinctive. C. pygmeaus
appears much smaller (9.0 mm in length) as
compared to C.parviceps (11.5 mm in length). The
tips of paraclypei appear separated in C. pygmeaus
but they clearly enclose clypeus in C. parviceps,
pronotum appears narrowish in C. pygmeaus as
compared to broader pronotum in C. parviceps,
lateral margins of pronotum appears distinctly
concave in the middle with posterolateral margins
distinctly sinuate in C. pygmeaus as compared to
lateral margins of pronotum slightly sinuate in C.
parviceps. We consider this species most closely
related with Linnavuori’s species C. mawambinus,
C. deserticolus and C. edentatus in having lateral
margin of abdominal venter less contrastedly pale
with brown punctures and body of small to medium
size but it appears entirely isolated in its clade in
having antennae short and incrassate, pronotum
very coarse and dense and paramere with apex of
blade subacute. We also have given here the key to
separate the above species to make the
identification of the presentspecies, much easier.
Linnavuori (1982) also gave some illustrations of C.
pygmaeus but his long key to the species of
Caystrus did not include only some of the above
species.
REFERENCES
AHMAD, I. (1986). A fool-proof technique for inflation
of male genitalia in Hemiptera (Insecta). J. ent.
Soc. Kar. 1 (2): 111-112.
AHMAD, I. AND MCPHERSON, J.E. (1990). Male
genitalia of the type species of Corimelaninae
White, Cydnoides Malloch and Galgupha Amyot
and
Serville
(Hemiptera:
Cydnidae:
coriomelaeniae)
and
their
bearing
on
classification. Ann. Entomol. Soc. Am. 83 (2):
162- 170.
AHMAD, I. AND MCPHERSON, J.E. (1998).
Additional information on male and female
genitalia of Parabrochymena Lariviere and
Brochymena Amyot and Serville (Hemiptera:
Pentatomidae). Ann. Entomol. Soc. Am. 91 (6):
800-807.
LINNAVUORI, R. (1972). Studies on African
Pentatomoidea. Arquivos do Museu Bocage (2)
3 (15): 395-434.
LINNAVUORI, R. (1974). Hemipterological studies.
Ann. Naturhist. Mus. Wien 78:393-413.
LINNAVUORI, R. (1982). Pentatomidae and
Acanthosomatidae (Heteroptera) of Nigeria and
the Ivory Coast, with remarks on species of the
adjacent countries in West and Central Africa.
Acta Zoologica Fennica, no. 163: 176 pp.
18
Zahid & Ahmed
STUDY OF RESISTANCE IN SITOPHILUS ORYZAE AGAINST BIOSAL,
CYPERMETHRIN AND PHOSPHINE ON THE BASIS OF TOXICITY VALUES
S.M. NAUSHAD ZAFAR1, TARIQ2, R.M. ASLAM3, M., BREUER4, M.,
NAQVI5, S.N.H. & PERVEEN6, R.
1
2
Department of Zoology, University of Karachi, Karachi-75270, Sindh-Pakistan.
M.A.H. Qadri Biological Research Centre, University of Karachi, Karachi-75270, Sindh-Pakistan.
3
Department of Department of Statistics, University of Karachi, Karachi-75270, Sindh-Pakistan.
4
Department of Ecology, Enology Section, Mehrhauser. Str. 119 Frieburg, Germany
5, 6
Department of Pharmacology, Baqai Medical University, Toll Plaza, Karachi, Sindh-Pakistan.
ABSTRACT
In the present work, the experiments were conducted for the resistance estimation in susceptible strain (SS),
Karachi strain (KS) and Lahore strains (LS) of Sitophilus oryzae L. (Rice weevil). The three strains (populations)
of S. oryzae were exposed to a neem formulation, Biosal (10 EC) available locally in the market, a synthetic
pyrethroid, Cypermethrin (10 EC) and a fumigant, Phosphine (PH3) used worldwide. The resistance was
estimated by filter paper impregnation contact method. The LD50 values were calculated by impregnated surface
area as μg/cm2.
The LC50 values of Biosal against three strains (populations) SS, KS & LS were computed as 18.600 47.1354
and 51.96 μg/cm2, respectively. Whereas the LC50 values of Cypermethrin against SS, KS and LS were
-4
-4
-4
2
computed as 29.08x10 , 40.40x10 and 44.33x10 μg/cm , respectively. While the LC50 values of Phosphine
(PH3) against SS, KS and LS were computed as 0.166429, 0.698113 and 0.84574 g/2.8m3, respectively.
Thus it may be concluded that, in three strains (populations), the increase in resistance may be shown as KS is
< LS. Whereas the toxicity (LD50) is in the following sequence, Biosal < Phosphine and < Cypermethrin.
As a whole the Karachi strain (KS) and Lahore strain (LS) were compared with that of susceptible strain and both
KS & LS were found more resistant to Cypermethrin and Phosphine. Lahore strain was found to be more
resistant as compared to Karachi strain, but least or negligible resistance was found against Biosal (neem
formulation = phytopesticide) which seems to be correct on the basis of LC50 values.
Key words: Resistance, Toxicity, S. oryzae, Biosal, Cypermethrin, Phosphine.
INTRODUCTION
Pakistan is an agricultural country; it depends
mainly on agricultural products. The cotton, paddy,
wheat and others are the main crops. Cereals have
an important nutritional value and are one of the
principal diet worldwide and among them rice has a
remarkable position. During storage, products may
be infested by insect pests causing deterioration of
the grain. This makes a treatment necessary for their
control. The rice weevils, Sitophilus oryzae (L.), the
granary weevil, Sitophilus granarius (Linnaeus), the
maize weevil, Sitophilus zeamais Motschulsky,
(Coleoptera: Curculionidae) and the broadnosed
grain weevil, Caulophilus oryzae (Gyllenhal) are
responsible for most of the insect-related damage of
stored rice and grain. These insects are called
primary insect pest, or internal feeder, because the
adults attack whole kernels and larvae feed and
develop entirely within the kernels. Oryzaephilus
surinamensis (L.) (Coleoptera: Silvanidae) is a
secondary pest that feeds on damaged kernels. The
Araecerus fasciculatus (De Geer) is 5th weevil which
is primary insect pest but is found in coffee beans.
To control stored grain pests different methods
have been used from time to time, e.g. by spray
method, fumigation method and biological method.
The fumigation method was more adopted as
compared to spray and biological control, due to
easy, effective and vast area covering control.
Indiscriminate and repeated use of the pesticides
and fumigants (Phosphine etc.) produced hazardous
effects specially, the resistance in the stored grain
pests. Many workers have reported resistance
against pesticide some of them are as follows (Stone
1968, W.H.O. 1970, Georghiou and Mellon 1983,
Saleem & Shakoori 1989, Preisler et al., 1990,
Zettler, 1991, Arthur and Zettler 1991, Chilcutt and
Tabashnik 1995, Grafius 1997, Mostafa and Zaved
1999, Fragoso et al., 2002, Asidi et al., 2005, Akiner
and Caglar 2006, N’Guessan et al., 2007, Khaliq et
al., 2007) they evaluated the resistance in stored
grain pest against pesticides.
Whereas resistance against fumigants, have
been reported by a number of researchers. Some of
them are as follows (Winks 1969, Tyler et al., 1983,
Taylor and Halliday 1986, White and Lambkin 1990,
Irshad and Iqbal 1994, Bengston et al., 1999,
Chaudhry 2000, Rajendram and Gunasekaran 2002,
20
S. M. N. Zafar et al.
Falaksher & Shakoori 2004. Amount of the fumigants
that may replace the ozone-depleting methyl
bromide, which is to be taken off the market, are
phosphine, ethyl formate, carbonyl sulfide, sulfuryl
fluoride, etc. The expected loss of methyl bromide
for the treatment of buildings, stored products and
soil is still regarded as a major problem, particularly
in the United States.
Fumigants are respiratory poisons and upon
exposure damage nervous system and muscles.
Lack of breathing may also cause accumulation of
CO2 which inturn keeps the spiracles open to allow
more fumigants to enter body cells as well as extra
cellular fluid and haemolymph. The acidosis due to
CO2 retention may effect the proteins and enzymes
in cells and haemolymph (Pratt and Reuss, 2004).
As it is well known that the use of persistent
organochlorines like DDT and the acute organophosphours compounds has led to hazardous effects
on environment and human beings. In response,
efforts were made to strengthen the Integrated Pest
Management (IPM) approach where there is
chemical control (Schmutterer 1995), if at all
necessary, should be combined with other methods
like crop rotation, resistant varieties and biological
control. In addition, attention was directed towards
the development of alternative chemicals.
The neem tree, Azadirachta indica A. Juss is so
far the most promising example of plants currently
used for pest control but Pruthi (1937) was the first
scientist who used neem as pesticide in India and
Siddiqui (1942) from Indo-Pak was the scientist who
isolated three bitter principles from neem oil for the
first time. This holy tree in Indo-Pak, from where it
originates, now has a global distribution throughout
the tropics. It is used for many purposes such as
shade tree, poles for construction, medicine, tooth
sticks and as a source of insecticide (National
Research Council, 1992). Since the early seventies
much research has been carried out on the
pesticidal properties of the neem tree and the results
have been published in proceedings from three
International Neem Conferences (Schmutterer and
Ascher, 1982; 1984; 1987). A summary of how neem
products are used as bio-pesticides, the mode of
action, effects on pests and natural enemies has
been reviewed by Schmutterer (1997). Although the
active ingredients in the neem, such as the
azadirachtin, are known, it has not been possible to
synthesize these complex compounds. Stable
formulations of purified extracts are commercialized
(Margosan-O and others) and distributed in several
countries.
The botanical pesticides prepared from neem
tree, other trees and plants are safer in use. It has
been proved that very little or no resistance develop
against phytopesticides, or developes after very long
time.
Naqvi (1987) discussed resistance in various
insect species due to indiscriminate use of pesticides
or constant use of the same pesticide in the field or
selection pressure in laboratory. These factors
increases the level of tolerance in the particular pest
strain to a great limit. The phenomenon is
biochemical and genetical. Hundreds of species and
strains have been reported to be resistant to various
pesticides or groups of pesticides. Sometimes crossresistance also develops.
Here in Pakistan the same problem of resistance
is being experienced by researchers and fumigators.
Therefore, the present work is a step to the solution
of this problem in Pakistan. Toxic effect of these
pesticides i.e – Phosphine (PH3), Pyrethroid
(Cypermethrin 10 EC) and Neem preparation
(Biosal) on S. oryzae were estimated. The work was
aimed to evaluate the level of resistance in S. oryzae
against these pesticides.
Pest collection and rearing technique
Stored grain insect pest Sitophilus oryzae adults
and its immature biological stages were collected for
experimental purpose. Samples were collected from
various grain warehouses at different localities of
Karachi i.e. various rice warehouses in Korangi,
Landhi, SITE area, TPX and Maripur Road. Insects
were collected separately in plastic bags along with
the rice on which they were feeding. They were
called as S. oryzae Karachi strain (KS). They were
then reared in the laboratory as a single population /
strain in University of Karachi.
Sitophilus oryzae Lahore strain (LS) were
collected from Rice warehouses in different localities
of Lahore and reared in the laboratory of SGS
Agricultural Laboratory, PECHS, Karachi under 28°C
+ 2°C temperature and at 65 + 5% r.h. Whereas
susceptible strain is being reared in the insectory of
Zoology Department, since 15 years. The susceptible
strain was given the code as (SS). Rearing of the
pest and its biological stages was done in the
insectory of Zoology Department, University of
Karachi. Insects were reared in glass jars filled with
rice in twelve (12) separate jars. Upper quarter of
each jar was left empty in order to provide space for
free movement of insects. Upper end of each jar was
covered by muslin cloth for ventilation and to avoid
escape of insects.
In order to avoid fungal attack due to high
humidity, grains were shaked and replaced by few
fresh grains from time to time and or whenever
required. Egg laying was allowed directly in the food
medium. Mature adults were introduced for this
purpose in the glass jars. Hatching of eggs was
observed regularly and required number of insects
were taken out and kept in the “Petri dishes” of 9
Study of resistance in Sitophilus oryzae against Biosal, Cypermethrin & Phosphine
Preparation of chemicals
The Biosal (10 EC) Neem formulation was
directly used in the present work, due to having low
toxicity against stored grain pest. Initially after trials
11.795 μg/cm2 to 27.522 μg/cm2 dose was selected
for susceptible strain and 15.726 μg/cm2 to 78.634
μg/cm2 dose was selected for both Karachi and
Lahore strain. Whereas of cypermethrin 0.1% stock
solution was prepared by dissolving 0.1 ml of
cypermethrin into 100 ml. bidistilled deionized water
and then different doses in ml were applied in the
experiment. After preliminary tests 0.15 ml to 1.00 ml
doses were used in the experiment and then
selected finalized doses were converted into μg/cm2
by using formula:
μg/cm2 = concentration x vol x 1000 x _1_
100
Area
While in the case of fumigant phosphine tablets
were used in the experiments. For this purpose from
0.125-2.00 tablets were kept in the artificial
fumigation chamber. This was done by breaking the
tablet into two parts and then in the four parts and
then it was weighed approximately as 0.125, 0.25
gm, 0.5 gm, 1.00 gm, and 2.00 gm.
Method of Treatment for Toxicity Determination:
A group of hundred (100), adult weevils having
uniform age and size was released in petri dishes
having 9 centimeter diameter (= 4.5cms radius) with
impregnated paper. The insects were released with
respective neem formulation (Biosal) and standard
(cypermethrin) having different concentrations. A set
of ten petri-dishes were set up for neem formulation,
in duplicate, nine for five different concentrations,
and one for control. For standard (cypermethrin)
also, six petri dishes were set up, nine for nine
different concentrations and one for control. Mortality
counts were made after 24 hours in each
experiment. Each experiment was repeated 5 times.
If in any experiment mortality rate increased more
than 10% in the case of control the experiment was
discarded. The observations were analyzed
according to Abbot’s formula (1925) and recorded in
the form of Tables.
Lethal Concentration (LC50) Values:
Average values were calculated and mortality
curve was drawn on log-log graph paper to find out
LC50 by taking dose on x-axis whereas the percent
mortalities on y-axis for Biosal, “Cypermethrin and
Phosphine following Raymond et al., (1993).
Statistical analysis was done on computer program
by fitting actual readings, which were transformed by
computer and the LC50 of Biosal, Cypermethrin and
Phosphine against three strains were calculated.
RESULTS
Toxicity
The Biosal 10 E.C (Neem formulation) was used
in the present work, due to its low toxicity against
stored grain pests especially S. oryzae. After
preliminary tests, nine concentrations were selected
for each strain i.e. for susceptible (SS), Karachi (KS)
and Lahore strain (LS). The concentrations of Biosal
in ml was selected for susceptible strain. LC50
determination doses were 1.500, 1.750, 2.000,
2.250, 2.500, 2.75, 3.00, 3.25 and 3.50ml/4.5cm2.
These concentrations were calibrated into 11.795,
13.761, 15.726, 17.692, 19.658, 21.624, 23.590,
25.556 and 27.522 μg/cm2 for quantification. The
mean mortalities % with S.D value caused by these
concentrations were 22 + 4.47, 28 + 8.36, 34 + 8.94,
42 + 4.47, 48 + 8.36, 56 + 5.48, 68 + 8.36, 82 +
4.47, and 96 + 8.94%, respectively.
Whereas for Karachi and Lahore strains, the
concentrations of Biosal selected for LC50
determination were 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0,
9.0 and 10 ml/4.5cm2. These ml concentrations were
them converted into 15.726, 23.590, 31.543, 39.317,
47.181, 55.044, 62.907, 70,771 and 78.634 μg/cm2.
The mean mortalities % with S.D value caused
by these concentrations in the case of Karachi strain
(KS) were 16 + 5.48, 20 + 7.07, 28 + 8.36, 36 + 5.48,
44 + 8.94, 52 + 4.47, 64 + 5.48, 78 + 8.36 and 96 +
8.94%, respectively.
While the mean mortalities % with S.D value
produced by these concentrations in the case of
Lahore strain (LS) were 14 + 5.48, 18 + 4.47, 26 +
5.48, 32 + 4.47, 40 + 7.07, 48 + 8.36, 58 + 8.36, 72
+ 4.47 and 88 + 8.36%, respectively.
The LC50 value against susceptible strain,
Karachi strain and Lahore strain of S. oryzae has
been found as 18.600, 47.135 and 51.96 μg/cm2,
respectively, and are shown in Fig. 1,2, and 3,
respectively.
Toxicity of Biosal (10 EC) against Sitophilus oryzae
Susceptible strain (SS) showing LC50 = 18.600 µg/cm2
Transform Mortality
cms diameter (= 4.5 cms radius) for treatment. In
order to determine the efficiency of insecticides and
fumigants artificial infestation was created in the
required commodity. Samples were then treated with
insecticides, i.e. Neem formulation (Biosal),
Cypermethrin and Fumigant (Phosphine).
21
1.5
1
0.5
0
-0.5 0
-1
-1.5
5
10
15
20
Concentrations
Figure 1
25
30
22
2
1
0
-1
0
20
40
60
80
100
-2
The LC50 values of cypermethrin against SS, KS
and LS were found as 29.08 x 10-4, 40.4 x 10-4 and
44.33 x 10-4 μg/cm2 as shown in Fig. 4, 5, and 6,
respectively.
Toxicity of Cypermethrin (10 EC) against Sitophilus
oryzae susceptible strain (SS) showing
LC50 = 0.002908 μg/cm2.
Concentrations
Figure 2
Toxicity of Biosal (10 EC) against Sitophilus oryzae
Lahore strain showing LC50 =51.96 μg/cm2
1.5
1.5
1
0
0.00E+00
-0.5
1.00E-03
2.00E-03
3.00E-03
4.00E-03
5.00E-03
-1
-1.5
Concentrations
Figure 4
1
0.5
-1
-1.5
20
40
60
80
100
Concentrations
Figure 3
Toxic effects of cypermethrin 10 EC (Pyrethroid)
were directly concentrations dependant. The
selected concentrations against the susceptible
strain (SS) of S. oryzae were 0.15, 0.20, 0.25, 0.30,
0.35, 0.40, 0.45, 0.50 and 0.55 ml / 4.5cm2. These
concentrations were then converted into, 11.795 x
10-4, 15.726 x 10-4, 19.658 x 10-4, 23.590 x 10-4,
27.522 x 10-4, 31.453 x 10-4, 35.385 x 10-4, 39.317 x
10-4 and 432.249 x 10-4 μg/cm2, respectively.
The mean mortalities percentage with S.D value
produced by these concentrations were 18 + 8.36,
26 + 7.07, 32 + 4.47, 44 + 8.94, 52 + 4.47, 66 +
5.48, 78 + 8.36, 84 + 5.48 and 96 + 8.94%,
respectively.
Whereas the selected ml concentrations used
against both for Karachi strain (KS) and Lahore
strain (LS) were 0.20, 0.30, 0.40, 0.50, 0.60, 0.70,
0.80, 0.90 and 1.00 ml / 4.5 cm2. These ml
concentrations were then converted into 15.72 x 104
, 23.59 x 10-4, 31.45 x 10-4, 39.31 x 10-4, 47.18 x 104
, 55.04 x 10-4, 62.90 x 10-4, 70.77 x 10-4 and 78.63 x
10-4 μg/cm2.
The mean mortalities % with S.D value
produced by these concentrations in the case of
Karachi strain (KS) were 24 + 5.48, 26 + 5.48, 34 +
8.94, 48 + 8.36, 56 + 5.48, 68 + 8.36, 76 + 5.48, 84
+ 5.48 and 96 + 8.94%, respectively.
Toxicity of Cypermenthrin (10 EC) against Sitophilus
oryzae L. Karachi strain (KS) showing
LC 50= 0.00404 μg/cm2.
Transform Mortality
0
-0.5 0
2
1.5
1
0.5
0
-0.5
0.00E+00 2.00E-03 4.00E-03 6.00E-03 8.00E-03 1.00E-02
-1
-1.5
Concentrations
Figure 5
Toxicity of Cypermentrin (10 EC) against Sitophilus
oryzae L. Lahore strain (LS) showing the
LC50 = 0.004433 μg/cm2
Transform Mortality
Transform Mortality
While by applying the above concentrations
same as in KS, the mean mortalities % with S.D
value shown against Lahore strain (LS) were 12 +
4.47, 24 + 5.48, 30 + 7.07, 44 + 5.48, 52 + 4.47, 64
+ 5.48, 70 + 7.07, 82 + 4.47 and 96 + 8.94%.
Transform Mortality
Transform Mortality
Toxicity of Biosal (10 EC) against Sitophilus
oryzae L. Karachi stain (KS) showing
the LC50 =47.1354 μg/cm2
2
1
0
0.00E+00
-1
-2
2.00E-03
4.00E-03
6.00E-03
8.00E-03
1.00E-02
Concentrations
Figure 6
Toxic effects of phosphine against S. oryzae L.
were noted at 5 different selected dose by applying
in the fumigation chamber constructed especially for
The dose of 0.0.125 g / 2.8 m3 caused no
mortality after 24 hours but after 48 hours the
mortality was 40% and after four days (96 hours)
100% mortality was noted. While at the dose of 0.25
g / 2.8 m3, 19% insects were killed after 24 hours
exposure and 60% mortality was noted after 48
hours and the same mortality was observed after
three days (72 hours). All the insects died after 96
hrs and showed 100%, mortality.
Moreover, 56% and 50% mortality was found at
0.5 g / 2.8 m3 after 24 hours in the case of Karachi
and Lahore strains, respectively. The fumigational
effects of above doses in the fumigation chamber
were noted after 24,48,72 and 96 hours. The 100%
mortality was found at 0.500 g / 2.8 m3, 1.00 gm,
2.00 gm, in susceptible strain while in the case of
Karachi and Lahore strains only at 1.00 and 2.00 g /
2.8 m3 100% mortality was found.
The LC50 of phosphine against three strains (SS,
KS and LS) was found to be 0.166429, 0.698113
and 0.84574 g/2.8m3, respectively as shown in Fig.
7, 8 and 9.
Transform Mortality
Toxicity of Phospine gas against S. oryzae L.
susceptible strain (SS) showing the
LC50= 0.166429 g/2.8m3
3
2
1
0
-1
0
0.5
1
1.5
Concentrations
Figure 7
2
2.5
Transform Mortality
The dose 0.125 g / 2.8m3 was applied at the
temperature of 24 + 2 oC and 66 + 2 r.h., while the
dose of 0.25 g / 2.8m3 was applied at the
temperature of 26 + 2 oC and 64 + 2 r.h. The dose of
0.50 g / 2.8m3 was applied at the temperature of 28
+ 2 oC and 62 + 2 r.h., whereas 1.00 g / 2.8 m3 dose
was applied at 30 + 2 and 60 + 2 r.h. The dose of
2.00 g / 2.8m3 was applied at the temperature of 32
+ 2 oC and 58 + 2 r.h.
Toxicity of Phosphine gas against Sitophilus oryzae L.
Karachi strain (KS) showing the
LC50 =0.698113 g/2.8m3
3
2
1
0
-1 0
0.5
1
1.5
2
2.5
-2
Concentrations
Figure 8
Toxicity of Phosphine gas against Sitophilus oryaze L.
Lahore strain (LS) showing LC50= 0.84574 g/2.8m3
Transform Mortality
this research work. The selected doses for treatment
of susceptible (SS) Karachi (KS) and Lahore strain
(LS) were 0.125, 0.250, 0.500, 1.00 and 2.00 g / 2.8
m3, respectively.
23
3
2
1
0
-1 0
0.5
1
1.5
2
2.5
-2
-3
Concentrations
Figure 9
DISCUSSION
In the present study, the experiments were
carried out for the estimation of resistance against
susceptible strain (SS), Karachi strain (KS) and
Lahore strain (LS) of Sitophilus oryzae L. (Rice
weevil) adult weevils. These strains were exposed to
a neem based formulation, Biosal (10 EC) locally
available in the market, a synthetic pyrethroid,
cypermethrin (10 EC) and a fumigant used
worldwide, phosphine (PH3). The resistance was
estimated on the basis of calculating the LC50 values
(contact method). Some other parameters may also
be used in this connection, as are being used in
different parts of the world.
Using the above parameter the following results
have been obtained which may be compared, with
the results obtained by other researchers and
workers. The results are as follows.
The LC50 values of Biosal against SS, KS and
LS were computed as 18.600 47.1354 and 51.96
μg/cm2, respectively. Whereas the LC50 value of
Cypermethrin against SS, KS and LS were
computed as 29.08x10-4, 40.4x10-4 and 44.33x10-4
24
μg/cm2, respectively. While the LC50 value of
Phosphine against three strains were computed as
0.166429, 0.698113 and 0.84574 g/2.8m3,
respectively. On the other hand if the pesticide and
fumigant are compared with each other, used in this
study, then the sequence of increasing resistance
may be shown as follows: Biosal < Phosphine and <
Cypermethrin, which means the least tolerance have
been found against the biopesticide.
Biosal falls in phytopesticide group whereas
Phosphine (fumigant) has shown greater resistance
in the present study. On the other hand
Cypermethrin showed the greatest resistance in the
present study, which belongs to the synthetic
pyrethroid group. The same type of study on
resistance was studied by Georghiou and Mellon
(1983). They studied the resistance among the
mosquitoes species, and reported that only among
the mosquitoes 96 species were found resistant to
one or more groups of pesticides. As there are five
groups
of
pesticide:
Carbamate
(CB),
Organophosphate (OP), Organochlorine (OC),
Synthetic pyrethroid (SP) and plant originated
Biopesticides (BP). Among these 36 species were
resistant to one group, 32 species to two groups, 19
species to three groups, 8 species to four groups
and only one species to all five groups of pesticide.
The last part of the above report is exactly in line
with the present study, in which least/negligible
resistance has been found against S. oryzae (SS) by
paper impregnation method.
Khan and Naqvi (1985) reported the
effectiveness and penetration of Phosphine gas
(PH3) by using a fixed dose of 30 grams = 10
tablets/1000 ft3 at temperature ranging 28 to 31oC
and humidities at 76-82%, upto the penetration of
4,8,12 feet and in paper bags of 4 kgs each, having
rice in them. Results indicated that 100% mortality of
cigarettes weevils was obtained at the rate of 10
tablets/1000 ft3 upto the 100 ft3 in small stocks,
godowns and warehouses, the minimum exposure of
48 to 72 hours or more was required depending
upon temperature, humidity and quality of grains to
be fumigated. In the present investigation it was
carried out against rice weevils instead of cigarette
weevils, using the same phosphine gas but on
different dose. In the above report 10 tablets/1000 ft3
gave 100% mortality upto 100 ft3 against cigarette
weevils in 48 to 72 hours, which mean 1 tablet/100
ft3, whereas in the present work 0.25 tablet/100 ft3
gave 100% mortality in 48 to 72 hours in susceptible
strain, which is 75% low dose as compared to above
report, while 0.5 tablet gave 100% mortality in rice
weevil in 48 to 72 hours against Karachi and Lahore
strains which is also 50% low as compared to above
report. The difference in LC50 value against
susceptible, Karachi and Lahore strains of S. oryzae
due to PH3 is because of different insects in the two
reports, but other parameters are in line.
Vollinger
(1987)
reported
comparative
experimental study for possible development of
resistance against neem seed kernel extract (NSKE)
and deltamethrin in two genetically different strains
of diamond back moth, the Plutella xylostella (L.)
upto 42 generations. Response to NSKE and to
deltamethrin was compared with that of untreated,
susceptible lines. The larvae were treated during 3rd
and 4th instar stage. NSKE-treated lines showed no
sign of resistance in feeding and fecundity tests,
while deltamethrin treated lines developed X20 and
X35 resistance. There was no cross-resistance
between deltamethrin, NSKE and diflubenzuron
(Dimilin, a synthetic molting inhibitor). Beside this,
activity of esterase enzyme did not change
significantly, over the 35 generations.
Similarly Naqvi and Tabassum (1992) reported
about probable resistance by experiments in the two
separate lines of Musca domestica (PCSIR-strain)
which were subjected to the pressure of neem
extract (RB-a) and cyfluthrin (Solfac 10% EC). Their
studies indicated that after 35 generations 12 fold
increase in LC50 of cyfluthrin occurred, whereas in
the case of RB-a slight increase was found, this
showed that possibly resistance against neem
fraction will not appear or will develop after a long
time.
Ahmad et al. (1998) reported the LC50 as
19μg/cm2 of Cypermethrin against S. oryzae
(Karachi strain) by paper impregnation method.
While in the present work it has been found 40.4x104
μg/cm2, which is many times high and show higher
resistance against Cypermethrin in last 15 years.
Ivania et al. (1998) have reported the higher
frequencies and high levels of resistance to
Phosphine (PH3) against S. oryzae population as
also detected by the Food and Agricultural
Organization (FAO), with 10% and 20%CO2. While
in the present work, higher resistance in S. oryzae
has been found against Phosphine gas without using
CO2 gas.
Swain and Baral (2004) treated Sitophilus
oryzae and Callosobruchus chinensis, with seven
plant species leaf powder, to compare efficacy as
pest control agent. They reported that Sitophilus
oryzae was reasonably controlled by neem leaf
powder and C. chinensis was controlled by begunia
leaf dust powder. In the present case the neem
product controlled the Sitophilus oryzae and lesser
resistance was found against this product as
compared to cypermethrin and phosphine as evident
from LC50 for SS, KS and LS strains (18.6, 47.1 and
59.6μg/cm2). It also indicated that phosphine and
cypermethrin are more toxic at a comparatively lower
dose.
Nighat et al (2007) reported resistance problem
against malathion and deltamethrin in C-strains of
Rhyzopertha dominica (C-Chichawatni; K-Karachi;
W-Wazirabad S-Sialkot; L-Lahore and M-Multan
strains), on the basis of LC50. They reported C-strain
most resistant to malathion (LC50 11.5 ppm) while Mstrain was most resistant against deltamethrin (LC50
10.55 ppm). They used mixture of the two against
different strains and reported affective against K, S,
and M-strain at 4.71, 5.35 and 8.38 ppm). In the
present study the level of resistance was determined
in KS and LS strains, based on LC50. Therefore, the
results are not exactly comparable. However, on the
basis of LC50 they confirm former findings.
REFERENCES
ABBOTT, W.S. (1925). A method of computing
effectiveness on insecticide. J. Econ. Entomol.
18: 265-267.
AHMAD, I., NAQVI, S.N.H., ALAM, T., AZMI, M.A.
AND TABASSUM, R. (1998). Toxicity of
cypermethrin and Acorus calamus extract on
Sitophilus oryzae. Proc. Pakistan Congr. Zool.
Vol. 18: 31-36.
AKINER, M.M. AND CAĞLAR, S.S. (2006). The
status
and
seasonal
changes
of
organophosphate and pyrethroid resistance in
Turkish populations of the housefly, Musca
domestica L. (Diptera: Muscidae). J. Vector.
Ecol. 31(2): 426-32.
ARTHUR, F.H. AND ZETTLER, J.L. (1991).
Malathion resistance in Tribolium castaneum
(Coleoptera:
Tenebrionidae):
Difference
between discriminating concentrations by topical
application and residual mortality on treated
surface.
J. econ. Entomol. 84: 721-726.
ASIDI, A.N., N’GUESSAN, R, KOFFI, A.A., CURTIS,
C.F., HOUGARD, J.M., CHANDRE, F.,
CORBEL, V., DARRIET, F., ZAIM, M. AND
ROWLAND, M.W. (2005). Experimental hut
evaluation of bednets treated with an
organophosphate (Chlorphyrifos-methyl) or a
pyrethroid (Lambdacyhalothrin) alone and in
combination
against
insecticide-resistant
Anopheles gambiae and Culex qeuinquefaciatus
mosquitoes. Malar. J. 4(1): 25.
BENGSTON, M., PATRICK, J., COLLINS,
GREGORY, J., DAGLISH, VIVIENNE, L.,
HALLMAN., KOPITTKE, R. AND PAVIC, H.
(1999). Inheritance of phosphine resistance in
Tribolium
castaneum
(Coleoptera:
Tenebrionidae).
Journal
of
Economic
Entomology 92(1): 17-20.
CHAUDHRY, M.Q. (2000). An introduction to
phosphine as a fumigant, and insect resistance.
Pesticide Outlook Issue 3, 88-91.
CHILCUTT, C.F., AND TABASHNIK, B.E. (1995).
Evolution of pesticide resistance and slope of
the concentration-mortality line: are they
related? J. Econ. Entomol. 88: 11-20.
FALAKSHER, ALI, S.S. AND SHAKOORI, A.R.
(2004). Phosphine induced changes in various
esterase levels in 4th instar larvae of Trogoderma
25
granarium. Pakistan J. Zool., Vol. 36(4): 257260.
FRAGOSO, D.B., GUEDES, R.N., PICANCO, M.C.
AND ZAMBOLIM, L. (2002). Insecticide use and
organophosphate resistance in the coffee leaf
miner Leucoptera coffeella (Lepidoptera:
Lyonetiidae). Bull. Entomol. Res. 92(3): 203-12.
GEORGHIOU, G.P. AND MELLON, R.B. (1983).
Pest resistance to pesticides. Edited by G.P.
Georghiou and P. Saito. pp. 1-66 and 769-792.
GRAFIUS, E. (1997). Economic impact of insecticide
resistance in the Colorado potato beetle
(Coleoptera: Chrysomelidae) on the Michigan
potato industry. J. Econ. Entomol. 90: 11441151.
IRSHAD, M. AND IQBAL, J. (1994). Phosphine
resistance in important stored grain insects in
Pakistan. Pakistan J. Zool., 26: 347-350.
IVANIA, A., RENATO, G.A.R., SCHEILLA, B., SILVA
V.R.T., CASTRO M. FERNANDA PENETADO,
M. DE. (1998). Effects of carbon dioxide and
phosphine mixtures on resistant populations of
stored-grain insects. Journal of Stored Products
Research 34(1): 27-32.
KHALIQ, A., ATTIQUE, M.N. AND SAYYED, A.H.
(2007). Evidence for resistance to pyrethroids
and organophosphate in Plutella xylostella
(Lepidoptera: Plutelliidae) from Pakistan. Bull.
Entomol. Res. 97(2): 191-200.
KHAN, S.A. AND NAQVI, S.N.H. (1985). Laboratory
tests on the efficacy and penetration of
phosphine at varied temperatures, humidities
and exposure periods against Lasioderma
serricorne Fabricius in coriander seeds
(Coleoptera: Anobiidae). Proc. Ent. Soc. Kar.
14-15: 59-66.
MOSTAFA, A.A. AND ZAVED, A.B. (1999).
Resistance of Musca domestica (L.) (Diptera:
Muscidae) from Gamasa city to some
insecticides. J. Egypt Socio. Parasitol. 29(1):
193-201.
N’GUESSAN, R., CORBEL, V., BONNET, J.,
YATES, A., ASIDI, A., BOKO, P., ODJO, A.,
AKOGBETO, M. AND ROWLAND, M. (2007).
Evaluation of indoxacarb, an oxadiazine
insecticide for the control of pyrethroid resistant
Anopheles gambiae (Diptera: Culicidae). J. Med.
Entomol. 44(2): 270-6.
NAQVI, S.N.H. (1987). Pesticide resistance in
dipterous insects and its position in Pakistan.
Pakistan J. Ent. Kar. 2(1): 59-66.
NAQVI, S.N.H. AND TABASSUM, R. (1992).
Probable development of resistance against
neem extract (RB-a) and cyfluthrin (Solfac 10%
EC), in Musca domestica L. (PCSIR strain).
Pakistan J. entomol. Karachi, Vol. 7(1-2): 9-16.
NATIONAL RESEARCH COUNCIL, (1992). Neem:
A Tree For Solving Global Problems. National
Academy Press, Washington, D.C. pp. 1-141.
26
NIGHAT, A.S., MUNIR, M., ALI, S.S. AND
SHAKOORI, A.R. (2007). Efficacy of mixtures of
an organophosphate malathion and a synthetic
pyrethroid deltamethrin against lesser grain
borer, Rhizopertha dominica. Pakistan J. of
Zoology. 39(3): 179-184.
PRATT, S.J. AND REUSS, R. (2004). Scrubbing
carbon dioxide prevents overestimation of insect
mortality in long-duration static phosphine
toxicity assays. Journal of Stored Products
Research 40, 233-239.
PREISLER, H.K., HOY, M.A. AND ROBERTSON,
J.L. (1990). Statistical analysis of modes of
inheritance for pesticide resistance. J. Econ.
Entomol. 83: 1649-1655.
PRUTHI, H.S. (1937). Report of the Imperial
Entomologist. Sci. Rep. Agric. Res. Inst., New
Delhi, pp. 1935-1936.
RAJENDRAM, S., GUNASEKARAN, N. (2002). The
response of phosphine resistant lesser grain borer
and rice weevil Sitophilus oryzae in mixed-age
culture to varying concentrations of phosphine.
Pest Management Science, 58: 277-281.
RAYMOND, M., PROTO, G. AND RATSIRA, D.
(1993). PROBIT analysis of mortality assays
displaying quantal response, Version 3.3,
Licence L 93019, Praxeme, 34680 St. Georges,
d’Orqeues, France.
SALEEM, M.A. AND SHAKOORI, A.R. (1989).
Toxicity
of
malathion,
permethrin
and
cypermethrin against resistant and susceptible
strain of Tribolium castaneum (Herbst.) in
Pakistan. Pakistan J. Zool. 21 (4): 347-360.
SCHMUTTERER, H., ASCHER, K.R.S. AND
REMBOLD, H. (1982). Natural pesticides from
the neem tree (Azadirachta indica A. Juss.).
Proc. 1st Intl. Neem Conf. (Rottach-Egern, 1980)
pp 1-297, Eschborn, 1982.
SCHMUTTERER, H. AND ASCHER, K.R.S. (1984). Natural
pesticides from the neem tree (Azadirachta indica A. Juss.)
and other tropical plants. Proc. 2nd Intl. Neem Conf.
(Rauischolzhausen, 1983) pp. 1-587, Eschborn, 1984.
SCHMUTTERER, H. AND ASCHER, K.R.S. (1987).
Natural pesticides from the neem tree
(Azadirachta indica A. Juss.) and other tropical
plants. Proc. 3rd Intl. Neem Conf. (Nairobi,
Kenya, 1986) pp 1-704, Eschborn, 1987.
SCHMUTTERER, H. (1995). The Neem Tree:
Source of Unique Natural Products for
integrated Pest Management, Medicine, Industry
and other purposes. VCH Weinheim - New York,
Basel. Cambridge. Tokyo 1-696 pp.
SCHMUTTERER, H. (1997). Side-effects of neem
(Azadirachta indica) products on insect
pathogens and natural enemies of spider mites
and insects. J. Appl. Entomol. 121(2): 121-128.
SIDDIQUI, S. (1942). A note on the isolation of three
new bitter principles from neem oil. Curr. Sci. 7:
266-273.
STONE, B.F. (1968). A formula for determining the
degree of dominance in case of monofactorial
inheritance of resistance to chemicals. Bull.
WHO 38: 325-326.
SWAIN, T.K. AND BARAL, K. (2004). Effect of
certain plant products on some stored grain
pests. Journal of Applied Zoological Research.
15(2): 229-231.
TAYLOR, R.W.D. AND HALLIDAY, D. (1986). The
geographical spread of resistance to phosphine
by coleopterous pests of stored products. pp. 607613. In [ed.]. Proceedings of the Crop Protection
Conference, Pests and Diseases, 17-20
November, 1986. Brighton, Metropole, England.
TYLER, P.S., TAYLOR, R.W. AND REES, D.P.
(1983). Insect resistance to phosphine
fumigation in food warehouses in Bangladesh.
Int. Pest Control 25: 10-13.
VOLLINGER, M. (1987). The possible development
of resistance against neem seed kernel extract
and deltamethrin in Plutella xylostella. Proc. 3rd
Int. Neem Conf. (Nairobi 1986), pp. 543-554.
WHITE, G.G., AND LAMBKIN, T.A. (1990). Baseline
responses to phosphine and resistance status of
stored-grain beetle pests in Queensland,
Australia. J. Econ. Entomol. 83: 1738-1744.
WINKS, R.G. (1969). Resistance to the fumigant
phosphine in a strain of Tribolium castaneum
(Herbst). Insect Toxicol. Info. Serv. 12: 178.
WORLD HEALTH ORGANIZATION (1970). Insecticide
resistance and vector control: 17th Report of WHO
Expert Committee on insecticides: WHO Tech.
Report, Ser. No. 443.
ZETTLER, J.L. (1991). Pesticide resistance in Tribolium
castaneum
and
T.
confusum
(Coleoptera:
Tenebrionidae) from flour mills in the United States. J.
Econ. Entomol. 84: 763-767.
EFFICACY OF IMEDACLOPRID AND ENDOSULFAN IN COMPARISON
WITH BIOSAL (BIOPESTICIDE) AGAINST MYZUS PERSICAE
(SULZER) ON MUSTARD CROP
MUHAMMED FAHEEM AKBAR1, NIKHAT YASMIN2, FARAH NAZ2 AND ABDUL HAQ3
1
Department of Agriculture and AgriBusiness Management, Karachi.
Deparment of Zoology, University of Karachi, Karachi-75270, Karachi-Pakistan.
3
Department of Food Science & Technology, University of Karachi, Karachi 75270, Pakistan.
2
ABSTRACT
Efficacy of different insecticides as foliar application was studied against mustard aphid Myzus persicae
(Sulzer) in Malir field (Karachi). The insecticides imidachloprid 25 WP (80 gramsl/acre), endosulfan (800
ml/acre) and Biosal (2000 ml/acre) were sprayed at one week interval using Knapsack hand sprayer
and a check plot was also maintained for comparison. Efficacy was sassessed by counting the aphid
mortality in mustard field plots. All tested insecticides performed better against aphid as compared to
untreated plots. Imidacloprid proved as the best with 86 percent aphid population reduction after 1st
nd
spray and 83 percent after 2 spray followed by Endosulfan and Biosal with 73, 70 and 56, 57 percent
reduction respectively.
Key words: Myzus persicae, Insecticides, Efficacy, Mustard.
INTRODUCTION
Next to Cotton and Rape, Mustard (Brassica
campestris) occupies the maximum area among
oilseed crops in Pakistan. Deficit of edible oil in
Pakistan is a chronic problem, so the
importance of brassica crop cannot be ignored.
Due to this importance the area under mustard
crop is increasing gradually from 227,000
hectares in 2005-06 to 256,000 hectares in
2006-07 (Source: Ministry of Food, Agriculture
and Cooperatives) with increasing trend in the
yield as well, which was 212,000 tonnes in
2006-07 as compared to 81,000 tonnes in
2005-06. One of major constraints in less
production of Brassica campestris is damage
due to insect pests. A number of insect pests
attack this crop including Myzus persicae
(Sulzer) which causes a major loss in crop yield
due to severe infestation in brassica. Over 250
species of the superfamily Aphidoidea feed on
agricultural and horticultural crops throughout
the world (Blackman & Eastop 1984). Aphids as
pests tend to have a wider host range than
economically unimportant species. The green
peach aphid, Myzus persicae, has an extremely
wide host range of over 100 plants including a
wide variety of vegetable and ornamental crops
(Baker 1982). Aphids suck sap from plant
(phloem) tissues using mouthparts modified for
piercing and sucking. Some aphids feed on
foliage while others feed on twigs, limbs,
branches, fruits, flowers or roots of plants.
Some species inject toxic salivary secretions
into plants during feeding. If left unchecked,
aphids can stunt plant growth, deform and
discolor leaves and fruit or cause gall formation
on leaves, stems and roots (Hamman 1985). In
case of heavy attack, plants wither resulting in a
drastic loss in seed yield and oil contents.
Keeping in view the importance of mustard crop
and pest infestation present studies were carried out
to control the aphid Myzus persicae (Sulzer) through
conventional and biopesticides to evaluate the
efficacy of Biosal which is environmentally friendly.
The experiment was conducted at the grower’s
field in Malir (Karachi) in a randomized complete
block design (RCBD) with three replicates having
four treatments including control plot. Three
insecticides viz. Imidachloprid 25 WP (80gm/acre)
Endosulfan 35 EC (800 ml/acre) and Biosal (2000
ml/acre) were tested.
After pest infestation, the crop was sprayed
twice at one week interval with hand operated
knapsack sprayer.
Pre-treatment data was collected before 24 hours of
each spray and post-treatment counts were made
after 24 hours, 72 hours and 168 hours. The
effectiveness of respective treatments was
computed through Henderson and Tilton formula
(1955). The data recorded was subjected to
statistical analysis using ANOVA and Duncan’s
multiple range Test (DMRT).
Akbar et al.
28
RESULTS
Results showed variation in relative toxicity and
persistence among all pesticide treatments. The
effectiveness however reduced with time interval in
case of Imedacloprid and Endosulfan, while Biosal
showed moderate result being less toxic initially,
gradually increasing after 72 hours with a decline in
effectiveness after 168 hours of each spray.
After 1st spray Imidacloprid was found to be
more toxic showing percent efficacy (90) after 24
(1st Spray)
Table 1
Insecticides
Dose
hours, (85) after 72 hours and (83) after 168 hours
followed by Endosulfan (81) after 24 hours, (72) after
72 hours and (66) after 168 hours, whereas Biosal
showed less toxic effect (51) after 24 hours, (61)
after 72 hours and (58) after 168 hours (Table 1).
Similar trend was observed after 2nd spray showing
effectiveness of Imedacloprid after 24, 72 & 168
hours as (88), (81) & (80), followed by Endosulfan
(76),(69) & (64) and biosal as (58),(60) & (55)
respectively, (Table 2).
% Efficacy After different Time Intervals
ml/Acre
24 Hours
72 Hours
Average
168 Hours
Imidacloprid
80 Grams
90
85
83
86
Endosulfan
800 ml
81
72
66
73
Biosal
2000 ml
51
61
58
56
8.7
LSD 0.05
(2nd Spray)
Table 2
Insecticides
Dose
ml/Acre
% Efficacy After different Time Intervals
24 Hours
72 Hours
Average
168 Hours
80 Grams
88
81
80
83
Endosulfan
800 ml
76
69
64
70
Biosal
2000 ml
58
60
55
57
Imidacloprid
7.5
LSD 0.05
Population Reduction
%age
Mustard
Aphid
DISCUSSION
100
80
60
Imidacloprid
40
Endosulfan
Azadirachtin
20
0
1 Day
3 Days
1st Spray
7 Days
1 Day
3 Days
2nd Spray
7 Days
Efficacy of Imidacloprid and Endosulfan in comparison with Biosal (Biopesticide) against Aphid
29
DISCUSSION
profenophos being moderate but yet more effective
than Azadirachtin.
According to the results, relative effectiveness of
the pesticides varied significantly after different time
intervals. Imedacloprid and Endosulfan were
comparatively more effective than biosal, which
being a neem based compound showed less toxic
effect may be due to strong antifeedant, insect
growth regulatory effect of neem (Azadirachta indica)
as referred by Mordue and Blackwell (1993). Similar
studies on relative efficacy of different insecticides
have shown variable results.
Higher persistence and effective control of
mustard aphid has been reported (Zheng et al. 1997)
by using carbofuran and Endosulfan under field
conditions (Begum et al. 1991, Brown et al. 1999).
Akbar et. al. (2006; 2007), evaluated the
effectiveness of Biosal (neem formulation) in
comparison with endosulfan and profenophos
against jassid on okra and brinjal at different time
intervals and found moderate effect of Biosal against
jassid while on okra profenophos found to be more
effective than endosulfan and vice versa on brinjal
crop against jassid.
Hussain et. al. (1992); Akbar et. al. (2005),
tested organophosphate(OPs) and pyrethroid
insecticides against white fly and jassid on soybean
and found OPs more effective than pyrethroids
against both insects; as pyrethroids lost their
effectiveness after 7 days while OPs were persistent
up to 14 days.
Naqvi et al. (2006) used Acorus Calmus (AC)
rhizome oil and Annona Squamosa (AS) seed oil
against sucking pests of cotton as compared to
Mospilan and Temaron. AC and AS were less
effective than Mospilan and Temaron but with higher
dose they gave 20-70% control of thrips, jassid and
white fly as compared to control plot.
Tariq et al., (2007) reported toxic effects against
cotton bollworms using AS and AC as botanical
pesticides in comparison with Karate, Polytrin-C and
Curacron and found as effective as standards even
better against Heliothis armigera after 2 weeks. In
case of spotted and pink boll worm AS and AC gave
moderate result as compared to Confidor + Deltafos
used standard. AS and AC seems less effective than
standards used but still AS and AC are safe to use
as eco-friendly biopesticides.
Narottam (2006) studied the effect of
endosulphan and azadirachtin when used alone
existed in middle order of effectiveness, whereas it
varied in efficacy when used in combination as
endosulphan + Bt (Bacillus thurigenesis) and
azadirachtin + Bt, however in either case azadiractin
was comparatively less effective than endosulfan.
Misrah (2002) also used similar combinations of
pesticides along with profenophos against aphids
and jassids of Okra and observed significant
superiority of some pesticides among others,
REFERENCES
AKBAR, M. F., YASMIN, N., KHAN,
QURESHI, M. H. AND NAZ, F.
Relative toxicity and persistence of
insecticides
against
jassid,
devastans dist. on soybean. Pakistan j.
Karachi, 20(1&2): 1-3
M. F.,
(2005).
different
amrasca
entomol.
AKBAR, M. F., YASMIN, N., KHAN, M. F., NAZ, F.
AND LATIF, T. A. 2006. Effectiveness of
profenophos in comparison with endosulfan
against amrasca devastans dist. on okra crop.
Pakistan j. entomol. Karachi, 21(1&2): 25-27
AKBAR, M. F., YASMIN, N., NAQVI, S. N. H.,
KHAN, M. F. AND NAZ, F. (2007). Relative
efficacy of biopesticide in comparison with
conventional pesticides against Amrasca
devastans dist. on brinjal crop. Pakistan j.
entomol. Karachi,22 (1&2):1-3
BAKER J. R. (1982). Insect and Related Pests of
Flowers and Foliage Plants. The North Carolina
Agricultural Extension Service. 75 pp.
BEGUM M., HUSSAIN M. AND TALUKDER F.A.
(1991) “Relative effectiveness of some granular
insecticides against mustard aphid”, Bang. J.
Agric. Sci., 18, 49-52.
BROWN J.J., MCCAFFEREY P., HARMON B.L.,
DAVIS J.B., BROWN A.P. AND ERICKSON
D.A. (1999) “Effect of late season insect
infestation on yield, yield components and oil
quality of Brassica napus, B. rapa, B. juncea and
Sinapis alba in the Pacific North West region of
theUnited States”, J. Agric. Sci., 132, 281-288.
HAMMAN P. J. (1985). Aphids on trees and shrubs.
L-1227. Texas Agricultural Extension Service
House and Landscape Pests. College Station,
Texas. 2 pp.
HENDERSON C.F. AND TILTON E.W. (1955). Tests
with acaricides against the brown wheat mite.
Journal of Economic Entomology 48:157-161.
HUSSAIN, T., KHAN, M. M., AKBAR, M. F., NAQVI,
S. M. S. H. AND RAJPUT, M. A. (1992). Relative
toxicity and persistence of different insecticides
against white fly, Bemisia tabaci Genn. on
soybean. Proc. Pakistan Congr. Zool., vol. 12,
pp. 295-297.
30
Akbar et al.
MISRAH, P. (2002). Field evaluation of some newer
insecticides against aphids (Aphis gossypii) and
jassids (Amrasca biguttula biguttula) on okra.
Ind. J. Entol. 64(1): 80-84.
MORDUE, A.J. AND BLACKWELL (1993).
Azadiractin: an update. J. Ins. Physiol., 39: 903924.
NAQVI, S. N. H., TARIQ, R. M., ZAFAR, S. M. N.,
AND ATTIQUE, M. R. (2006). Efficacy of Acorus
calamus (AC) rhizome oil and Annona
squamosa (AS) seed oil against sucking pests of
cotton at CCRI-Multan (Punjab) as compared to
Mospilan and Temaron. Pakistan j. entomol.
Karachi,21 (1&2):23-27.
NAROTTAM, K. M. (2006). Evaluation of some new
insecticides and biopesticides against jassid,
Amrasca
biguttula
biguttula
on
okra,
abelmoschus esculentus. Ind. Jor. App.
Entol.20: 1.
TARIQ, R. M., NAQVI, S. N. H., ZAFAR, S. M. N.
AND BURRERO, A. S. (2007). Toxic effects of
botanical pesticide, from Acorus calamus(AC)
and Annona squamosa (AS) against bollworms
at ARI-Tandojam, Sindh-Pakistan, Pakistan j.
entomol. Karachi,22 (1&2):31-36.
ZHENG B.Z., GAO X.W., ZHAO G.Y. AND CAO B.J.
(1997) “Insecticide resistance in turnip aphids,
Lipaphis erysimi (Kaltenbach), from Beijing and
suburbs”, Resis. Pest. Mngt., 9, 27-28.
REVISION OF THE GENUS ISOMETRUS HEMPRICH & EHRENBERG
(SCORPIONIDA: BUTHIDAE: CENTRURINAE) WITH DESCRIPTION OF
TWO NEW SPECIES AND CLADISTIC RELATIONSHIP FROM PAKISTAN
RAFAT AMIR AND SYED KAMALUDDIN
Department of Zoology Govt. Degree Girls College, 11-B, North Karachi. (R.A),
Federal Urdu University of Arts, Science and Technology
Gulshan-e-Iqbal Campus (S.K)
ABSTRACT
The genus Isometrus Hemprich & Ehrenberg is revised to accommodating two new species from Pakistan with
special reference to their morphological characters, male genitalia, electrophoresis and gel filtration
chromatography of the venom. A key to the sub-genera and 12-species of the genus Isometrus is also
formulated. The new species are also compared to their closest allies.
Key Words: Isometrus, New species, Buthidae, Key, electrophoresis, chromatography, cladistic relationship,
Pakistan.
INTRODUCTION
The representatives of the genus Isometrus are
disturbed in Tropical areas of the world like India,
Pakistan, Srilanka, Burma, Java, Australia, Africa,
South America and South Sea Islands. Both new
species are recorded from Pakistan, Sindh.
Pocock (1900) has been described the genus
Isometrus, later Vachon (1969 & 1972) worked on
Isometrus. Tikader and Bastawade (1983) have
been described the genus Isometrus from India and
proposed two genus on the basis of trichobothrial
pattern on pedipalpi. They also formulated a key to
the sub-genera and a key of nine species from India.
(1972), Cah. Pacif, 16: 169-180; Stahnke (1972),
Ent. News. 83: 121-133.
Diagnostic features:
Carapace sometimes with only median carina,
mesosomal tergites monocarinated, inferior surface
of cheliceral fixed finger with one tooth,
cephalothracic sternum small, triangular and not as
long as genital operculum, caudal region slender,
vesicle as wide as metasomal segment V and with a
strong, triangular subaculeus spine at the base of
aculeus. Pedipalpi tarsus (movable finger) with 5-6
distinct median oblique rows of granules (nonimbricated) and a short apical depressed and fringed
with setae. Trichobothria dorsal 1, dorsal 3 and
dorsal 4 on femur from β angle.
Comparative note:
The animals were collected from the Korangi,
Sindh Pakistan and were killed with the help of
formalin and preserved in 70% Alcohol.
For the sudy of male genitalia the specimens
were dissected out by removing the tergites of
mesosoma. After dissection the aedeagus was
mounted on slide then taken the photograph using
microscope and photographic camera. For the study
of electrophoresis and chromatography the
technique generally followed by Amir et al. (1994
a,b,c,d, 1995, 2003 and 2004).
RESULTS
Genus: Isometrus Hemprich and Ehrenberg
Ismetrus Hemprich and Ehrenberg (1829). Phys.
Scorp., 3; Kraepelin (1891), Jb. Hamb. Wiss. Anst. 8:
244; Pocock (1900), Fauna Brit. India, Arachn., 4445; Vachon (1969), Senckenberg Biol. 50: 417-420;
The genus Isometrus is the only genus of the
sub-family Centrorinae Kreplain. The members of
this genus can easily be separated in having
mesosomal tergites monocarinated, inferior surface
of cheliceral fixed finger with one tooth, trichobothria
dorsal 1, dorsal 3 and dorsal 4 on femur from β
angle and by the other characters as noted in the
description.
Key to the Subgenera and species of the genus
Isometrus Hemprich and Ehrenberg.
1. Trichobothria db placed always proximal to et
but distal to est, vesicle is not bulbous...I.
Raddyanus Vachon............................................2
-
Trichobothria db placed always distal to et, vesicle is more
bulbous..................................................................................I.
Closotrichus.........................................................................11
32
Rafat Amir & Syed Kamaluddin
segments II and III may or may not be ending
posteriorly into an upstanding procurved
spines, sub-aculeus spine provided with three
pairs of small denticulate granules on inner
margin..............................................................10
2. Metasoma 5 to 6 times as long as carapace,
vesicle
twice
as
long
as
aculeus...............................................................3
-
Metasoma 7 to 8 times as long as carapace,
vesicle more than twice as long as
aculeus...............................................................8
-
3. Metasoma always 5 times as long as carapace,
sub-aculeus spine provided with one pair of
small
denticulate
granules
on
inner
margin................................................................4
-
Metasoma always 6 times as long as carapace,
sub-aculeus spine provided with one or three
pairs of small denticulate granules on inner
margin................................................................5
4. Mesosomal tergites with 5 black and 6 yellow
bands,
vesicle globular and as long as
wide……………………......I. (R) rigidulus Pocock
-
Mesosomal tergites with a pair of conspicuous
median yellow bands, vesicle elongated and
longer than wide.................................................6
10. Ratio of median eyes to anterior and posterior
margins of carapace 1:1.9, vesicle tapering
distally and much granules on ventral surface,
fingers of pedipalpi short and stout, trichobothria
ei placed proximally to d5 on femur, dorsal
carinae of metasomal segments II and III ending
posteriorly into very strong, procurved
and
upstanding spines….... /. (R) acanthurus Pocock
-
5. Entire surface very weakly and finely granular,
pectinal teeth 12-13............ /. (R) vittatus Pocock
-
Entire surface of carapace weakly granular, 32
pectinal teeth...........................I. (R) atherii sp. n.
6. Subaculeus spine provided with only one pair of
small denticulate granules on inner margin,
manus
of
pedipalpi
smooth
and
acarinated...........................................................7
-
Subaculeus spine provided with three pairs of
small denticulate granules on inner margin,
manus of pedipalpi granular
and strongly
carinated..................I. (R) brachycentrus Pocock
7. Pectinal teeth 15/16, body size more than 35
mm in length, body colour yellowish variegated
with blakish brown patches, manus of pedipalpi
short, stout in male as well as in
female.............................. I. (R) thrustoni Pocock
-
Pectinal teeth 11/12, body size not more than 30mm
in length, body color brownish variedated with yellow
patches, manus of pedipalpi very long in
male.................. I. (R) isadensis Tikader & Bastawade
8. Metasoma 8 times as long as carapace, subaculeus spine provided with one pair of small
denticulate granules on inner margin, manus of
pedipalpi very long and thin in male, pectinal
teeth 17/17............................ /. (R) europaeus L.
-
Metasoma 7 times as long as carapace, subaculeus spine provided with two or three pairs
of small denticulate granules on inner margin,
manus of pedipalpi short and stout in male,
pectinal teeth less than 17/17.............................9
9. Aculeus very short and less than one third of
vesicular length, dorsal carinae of metasomal
Aculeus slightly longer and less than half the
length of vesicle, dorsal carinae of metasomal
segments II and III posteriorly spiniform, subaculeus spine provided with two small
denticulate
granules
on
inner
margin………………...............assamensis Oates
Ratio of median eyes to anterior and posterior
margins of carapace 1:2.25, vesicle globular on
distal portion and very weakly granular on ventral
surface, fingers of pedipalpi thin and long,
trichobothria e1 placed in a same plane to d5 on
femur, dorsal carinae of metasomal segments II and
III ending posteriorly into a very small sub-denticular
granules……...I. (R) corbeti Tikader and Bastawade
11. Body variegated with black to brown
bands and spots, entire surface coarsely
granular, pectinal teeth 15-16….…….I. (C)
sankeriensis Tikader and Bastawade
-
Body generally mustered yellow, entire surface
granular, pectinal teeth 29….. I (C) liaqatii sp. n.
Isometrus (Raddyanus) atherii sp.n.
(Figs. 1-11)
Colouration: Body generally yellow.
Prosoma (Fig.3): Entire surface of carapace weakly
granular, all carinae granular, ocular tubercles
reddish brown and granular, anterior margins
smooth and provided with 30-33 small reddish setae,
lateral margins granular but more granular on
anterior portion.
Pedipalp (Figs. 4a-4c): Manus stout, longer than
femur, shorter than carapace, almost all carinae
strongly granular and outer and anterior side
provided with a crenulated crest, 16-19 denticular
tubercles, patella longer than femur but shorter than
carapace, carinae on outer side granular, inner or
anterior surface provided almost granular strong
crest with 16 sub-denticular tubercles, manus or
hand flat and stout, length of underhand longer than
femur, fixed finger almost as long as femur but
Revision of the genus Isometrus Hemprich & Ehrenberg (Scorpionida: Buthidae: Centrurinae)
movable finger longer than carapace, dentition on
the fingers consisting of two rows of imbricated
teeth, granular on the fixed and movable finger,
trichobothrial pattern of pedipalp 'A' type.
Legs (Fig 5): Femur, patella smooth and carinae
crenulated, tibiae with strong tibial spurs, on the legs
III and IV, size 0.10 cm., pedal spurs spiny,
tarsomere I laterally smooth, tarsomere II smooth
and furnished below with one row of bristles on
ventral surface.
Pectin (Fig. 7): Pectin well developed and almost
two times longer than wide, seven middle lamellae
present, fulcra nearly triangular, pectin pale yellow
with 32 teeth.
Genital operculum (Fig. 8): Genital operculum
wider than long and sclerites slightly divided on
posterior portion from which small genital papillae
produced in male, sternum small and triangular.
Mesosoma: All tergites more smooth on posterior
portion of each tergite, sternites I-IV smooth and
each provided with slit-like stigmata for book lungs.
Metason: Cauda four times as long as carapace,
first segment shorter than wide, segments I-IV with
dorsal carinae crenulated, dentiform on posterior
portion, much more elevated on segment III and IV,
dorsolateral carinae evenly crenulated, lateral
carinae weakly developed only on posterior portion
of segment III-IV.
Telson (Fig. 6): Telson with vesicle not as wide or
deep as segment V, ventral surface densely smooth,
ventral median crest developed, sub-aculeus nodule
absent, aculeus weakly curved, as long as vesicle.
Male genitalia (Fig. 9): Flagellum 0.71 mm long,
elongated, elastic and flageller-like, trunk 0.52 mm
long, 0.12 mm wide, trunk cylindrical basally dilated,
pedicel very flat, 0.2 mm long, 0.1 mm wide, sperm
spine blunt, sperm tube narrowed sclerotized.
Material examined: Holotype, Male, Pakistan,
Shikarpur (Sindh), 7.12.93, leg. Rafat Amir, lodged
at MEMUK No. 86.
Paratypes: 7 females, other data same as holotype,
lodeged at ZMUK.
Comparative note: This new species is most
closely related to Isometrus (Raddyanus) vittatus
(Pocock) in having mesosomal tergites with a pair of
conspicuous median yellow bands, vesicle elongated
and longer than wide but it can easily be separated
from the same in having entire surface of carapace
weakly granular, 32 pectinal teeth are present and
33
by the other characters as noted in the key and
description.
Isometrus (Closotrichus) liaqatii sp.n.
(Figs. 12-22)
Colouration: Body generally mustered colour.
Prosoma (Fig. 14): Entire surface of carapace
granular, all carinae weakly granular, ocular
tubercles dark brown and black, anterior margins
smooth and provided with 23-44 small brownish
setae, lateral margins crenulated on anterior portion.
Pedipalp (Figs. 15a-15c): Manus slender shorter
than femur and carapace, almost all carinae weakly
granular, outer and anterior side provided with a
crenulated crest of 23-25 denticular tubercles,
patella longer than femur but always shorter than
carapace, inner or anterior surface provided almost
granular crest with 22 sub-denticular tubercles,
manus or hand slender and length of underhand
longer than femur, fixed finger almost as long as
femur but movable finger shorter than carapace,
dentition on the fingers consisting of three rows of
imbricated teeth, granular on the fixed and movable
finger, trichobothrial pattern of pedipalp 'A' type.
Legs (Fig. 16): Femur weakly granular, patella
smooth and carinae crenulated, tibiae with two
strong tibial spurs on the legs III and IV, size 0.1 cm,
pedal spurs spiny, tarsomere I laterally smooth, a
pair of pedal spurs present, tarsomere II flanked
below, distal portion with a few bristles.
Pectin (Fig. 18): Pectin well developed and almost
two and one fourth times longer than wide, eight
middle lamellae present, fulcra nearly triangular,
fulcra and lamellae clothed with microscopic hairs,
pectin light yellow with 29 teeth.
Genital operculum (Fig. 19): Genital operculum
wider than long and sclerites slightly divided on
posterior portion from which small genital papillae
produced in male, sternum small and triangular.
Mesosoma: All tergites granular but more granular
on posterior portion of each tergite, sternites I-V
granular and each provided with slit-like stigmata for
book lungs.
Metasoma: Cauda four times as long as carapace,
first segment shorter than wide, segments I-IV with
dorsal carinae crenulated, dentiform on posterior
portion, much more elevated on segment III and IV,
dorso-lateral carinae evenly crenulated, lateral
carinae strongly developed only on posterior protion
of segment III-IV.
34
Telson (Fig. 17): Telson with vesicle not deep as
segment V, ventral surface densely granular, ventral
median crest not developed, sub-aculeus nodule
absent, aculeus not curved, as long as vesicle.
Male genitalia (Fig. 20): Flagellum 0.6 mm long,
membranous, trunk cylindrical and blunt, 0.4 mm
long and 0.15 mm wide, sperm spine typically blunt,
sperm tube narrowed and short.
Material examined: Holotype, Male, Pakistan:
Korangi, (Karachi), 9.7.1993, leg. Rafat Amir, lodged
at MEMUK No. 148.
Paratypes: 19 females, other data same as
holotype, lodged at ZMUK.
Comparative note: This new species is most
closely related to Isometrus (Closotrichus)
sankeriensis in having entire trichobothria db placed
always distal to et and vesicle is more bulbous but it
can easily be separated from the same in having
entire surface of carapace granular, 29-pectinal
teeth are present and by the other characters an
noted in the key and description.
Polyacrylamide Gel Electrophoresis of Scorpion
Venom
Polyacrylamide gel electrophoresis of scorpion
venoms were performed under denaturing condition
using SDS-70L standard protein markers were used
to prepare standard curve. The marker include
bovine serum albumin (66.0 KDa), egg albumin (45.0
KDa), glyceraldehydes β phosphate, dehydroginase
(36.0 KDa), carbonic anhydrase (29.0 DKa),
trypsinogen (24.0 KDa), trypsin inhibiter (20.1 KDa),
and X- lactalbumin (14.2 KDa). All venom samples
showed multiple bands in the molecular weight
range of (70.0 to 14.0 KDa). Venom of Isometrus
species (Figs. 10 & 21) showed few broad bands.
Gel filtration
venom:
chromatography
of
scorpion
1000 mg of each of two Isometrus atherii sp.n.,
and /. iqbalis Sp,n., venoms were loaded on
sephadex G-50 column (2.5x90.0 cm) and eluted
with 0.1 M ammonium acetate buffer (Ph 6.90). Both
venoms were resolved into four peaks, representing
the presence of high to moderate molecular weight
components. Isometrus atherii (Fig. 11) posses two
major (I and III) peaks, whereas Isometrus liaqatii
(Fig. 22) into three major (I to III) and one moderate
peak (IV).
TABLE 1
Measurement in cm/mm meristic characters of the
male holotype Isometrus (Raddyanus) atherii sp.n.
Characters
Total length
Carapace length
Mesosoma length
Metasoma length
I segment length/wide
II segment length/wide
Ill segment length/wide
IV segment length/ wide
V segment length/wide
Telson length
Vesicle length/wide
Aculeus length
Pedipalp length
Femur length/wide
Patella length /wide
Chela length/wide
Fixed finger length
Movable finger length
Chelicera, Chela length/wide
Fixed finger length
Pectinal tooth count
Male genitalia
Holotype Male
4.5 cm.
0.6 cm.
1.4 cm.
2.5 cm.
0.4/0.26 cm.
0.45/0.30 cm.
0.5/0.33 cm.
0.55/0.36 cm.
0.6/0.4 cm.
0.6 cm.
0.3/0.3 cm.
0.3 cm.
1.7 cm.
0.5/0.2 cm.
0.6/0.3 cm.
0.9/0.3 cm.
0.5 cm.
0.52 cm.
0.2/0.1 cm.
0.11 cm.
0.12 cm.
32
1.25 mm
TABLE 2
Variation in tarsomere II spine counts in Isometrus
(Raddyanus) atherii sp.n. on each specimen, the
spine of the left and right legs of each pair were
counted
Legs
Margin
4
5
6
7
8
I
Prolateral
5
8
4
7
5
Retrolateral
5
7
6
7
7
Prolateral
4
4
6
9
5
Retrolateral
5
5
6
8
6
Prolateral
5
7
5
4
6
Retrolateral
6
6
5
5
7
Prolateral
5
5
5
7
5
Retrolateral
5
5
4
4
7
II
III
IV
35
TABLE 3
Measurement in cm/mm meristic characters of the
male holotype Isometrus (Closotiichus) liaqatii sp.n.
Characters
Total length
Carapace length
Mesosoma length
Metasoma length
I segment length/wide
II segment length/wide
Ill segment length/wide
IV segment length/ wide
V segment length/wide
Telson length
Vesicle length/wide
Aculeus length
Pedipalp length
Femur length/wide
Patella length /wide
Chela length/wide
Fixed finger length
Chelicera, Chela length/wide
Fixed finger length
Pectinal tooth count
Male genitalia
Holotype Male
4.6 cm.
0.65 cm.
1.2 cm.
2.76 cm.
0.50/0.2 cm.
0.53/0.22 cm.
0.55/0.25 cm.
0.58/0.27 cm.
0.6/0.3 cm.
0.6/0.3 cm.
0.3/0.3 cm.
0.3 cm.
1.7 cm.
0.45/0.2 cm.
0.5/0.25 cm.
0.85/0.25 cm.
0.45 cm.
0.50 cm.
0.35/0.2 cm.
0.13 cm.
0.15 cm.
29
1.3 mm.
DISCUSSION
The genus Isometrus Hemprich and Ehrenberg,
is an only genus of the sub-family Centrurinae
Kraepelin, and distributed in Oriental, Australian,
Ethiopean and Neotropical regions. The sub-family
Centrurinae plays sister group relationship with
Buthinae by their synapomorphies like dorsal arm of
movable fringes of chelicerae furnished with four
minute teeth on inner margin and trichobothrial
pattern is A-type, but plays out group relationship by
its autapomorphy III and IV pairs of leg without tibial
spur.
Presently (Fig. 23) the genus Isometrus includes
twelve species which fall into two groups. The first
group includes only two species viz. sankeriensis
and liaqatii which plays sister group relationship with
each other and out group relationship with second
group, which includes ten species viz. rigidulus,
vittatus, atherii, bradycentrus, thrustoni, isadensis,
europaeus, corbeti, acanthurus and assamensis.
The second group further fall into two sub-groups.
Among first sub-group the corbeti and acanthurus
play sister group relationship with assamensis,
further all these three species play sister group
relationship with each other and out group
relationship with europaeus.
TABLE 4
Variation in tarsomere II spine counts in Isometrus
(Raddyanus) liaqatii sp.n. on each specimen, the
spine of the left and right legs of each pair were
counted
Among second subgroup, the thrustoni plays
sister-group relationship with isadensis and out
group relationship with bradycentrus. All these play
sister group relationship with rigidulus, vittatus and
atherii play sister group relationship with each other
and out group relationship with rigidulus.
ILLUSTRATION OF FIGURES
Legs
Margin
4
5
6
7
8
I
Prolateral
6
4
5
4
4
Retrolateral
5
5
5
5
5
Prolateral
6
3
5
7
5
Retrolateral
4
3
4
5
3
Prolateral
7
5
5
5
6
Retrolateral
4
4
7
4
3
Prolateral
7
6
8
5
5
Retrolateral
5
3
4
4
4
II
III
IV
Figs. 1-11. Isometrus atherii (sp. n.): 1. entire, dorsal
view; 2. same, ventral view; 3. prosoma, dorsal view;
4a-4c. pedipalp, lateral view, a. femur, b. patella, c.
hand; 5. leg, lateral view; 6. telson, lateral view; 7.
pectin, ventral view; 8. genital operculum, ventral
view; 9. aedeagus, lateral view, 10. showing
electrophoresis
of
venom;
11.
showing
chromatography of venom. Figs. 12-22. Isometrus
liaqatii (sp.n.): 12. entire, dorsal view; 13. same,
ventral view; 14. prosoma, dorsal view; 15a-15c.
pedipalp, lateral view; a. femur, b. patella, c. hand;
16. leg, lateral view; 17. telson, lateral view; 18.
pectin, ventral view; 19. genital operculum, ventral
view; 20. aedeagus, lateral view; 21. showing
electrophoresis
of
venom;
22.
showing
chromatography of venom.
Fig. 23. Cladogram showing relationship of the
included taxa.
36
37
38
39
40
REFERENCES
AMIR, R., ALAM. J.M. AND KHAN, M.A.J. (l994a).
comparative study of two toxins with
phospalipase A^ activity isolated from the
venom of Androctonus australis. Pakistan J.
Zool. 26 (2): 127-133.
AMIR, R., ALAM. J.M. AND KHAN, M.A.J. (l994b).
Investigation on scorpion venoms as naval
insecticides. Pakistan J. entomal Karachi, 9:
109-114.
AMIR, R., ALAM. J.M. AND KHAN, M.A.J. (l994c).
proloagulent property of venoms of some
medically important scorpion from Sindh
region. Pakistan J. Zool. 26(3): 261- 263.
AMIR, R., ALAM. J.M. AND KHAN, M.A.J. (l994d).
Comparative studies on the enzymatic content
of venom from fifteen scorpion species from
Sindh region. Pakistan J. Zool. 26 (1): 77-79.
AMIR, R., ALAM. J.M. AND KHAN, M.A.J. (1995).
Insecticidal activity of Toxic components from
scorpion venoms and their effects on cationic
concentration of Haemalymph of Periplanata
Americana. Pakistan j. entomol. Karachi,
70:45-49.
AMIR, R., KAMALUDDIN, S. AND KHAN, M.A.J.
(2003). Redescription of Androctonus sp.
(Sorpionida: Buthidae) from Sindh, Pakistan
with special reference to its genitalia and
chemical analysis of venom. J. nat. hist. wildl.
2 (2): 21-25.
AMIR, R., KAMALUDDIN, S. AND KHAN, M.A.J.
(2004). Redescription of Odontobuthus doriae
odonturus (Pocock), (Arachnida: Scorpionida:
Buthidae) from Pakistan with special reference
to
its
genitalia,
chromatography
and
electrophoresis of venom. J. nat. hist. Wildl. 3
(1): 17-21.
POCOCK, R. L. (1900). Fauna of British India,
Arachnida, London: 1-279.
STAHNKE, H. I. (1972). A nomenclatural
conundrum. Ent. News. 83:121-133.
TIKADER, B.K. AND BASTAWADE, D.B. (1983).
The fauna of India. Scorpion (Scorpionida &
Arachnida). Zool. Ind., 3: 1-671.
VACHON, M. (1969). Complement a la description
C’ Isometrus madagassus Roewer 1943
(Scorpiones: Buthidae). Sencken-berg. boil. 50
(5-6): 417-420.
VACHON, M. (1972). Sur l’ establissement d'une
nomenclature
trichobothriale
uniforme
convenout existence de trios types distinct de
trichobothriotaxie, C. r. hebd. Seunc. Acad.
Sci. paris, 275D (18): 2001-2004.
TAXONOMIC STUDIES OF SERGENTOMYIA BAGHDADIS (ADLER AND
THEODOR) (DIPTERA: PSYCHODIDAE) IN SINDH AND PUNJAB AND
COMPARISION WITH OTHER PAKISTANI SPECIES OF
THE SUBGENUS PARROTOMYIA THEODOR
JUMA KHAN KAKARSULEMANKHEL
Investigator of Sandflies, Leishmaniases, Helminths, Ticks & Mosquitoes, Department of Zoology,
University of Balochistan, Saryab Road, Quetta, Pakistan.
[email protected]
Cell # 0333-7860240
ABSTRACT
During the routine sandfly survey, Sergentomyia (Parrotomyia) baghdadis (Adler and Theodor) was
collected for the first time from new epidemic localities of cutaneous leishmaniasis in Sindh and Punjab
In view of the published reports about the detection of encephalitis viruses from the species of the
genus Sergentomyia Franca and Theodor, from the Indian localities, the correct identification of sand fly
species, becomes of significant value in the study of epidemiology of leishmaniases and other viral
diseases. Therefore, in order to facilitate Zoologists and Medical researchers in correct identification,
taxonomic characters of S. baghdadis were studied in detail with special reference to its mouth parts,
male and female genitalia and findings are presented in the present paper. A comparative note of
taxonomic characters of this species with its closest allies has also been given.
Key words: Sandflies, Sergentomyia baghdadis, Parrotomyia, Sindh, Punjab.
INTRODUCTION
Pakistan has several endemic forms of
leishmaniases, a protozoan disease transmitted by
the bite of infected sand flies. The disease is
spreading continuously and sand flies are being
recorded from new localities.
Lewis (1967,1978) reported the prevalence of
five species of sand flies of the subgenus
Parrotomyia Theodor of the genus Sergentomyia
Franca and Theodor in the country viz. S. africana
(Sinton), S. shorttii (Adler and Theodor), S. grekovi
(Khodukin) and S. palestinensis (Adler and Theodor)
including the present one. However, S. grekovi, S.
palestinensis and S. baghdadis (Adler & Theodor)
have also been reported from the Balochistan
Province (Kakarsulemankhel, 2004,2006).
Previously, several viruses have been found in
sand flies (Lewis, 1978). In view of the recently
published reports about the detection of encephalitis
viruses from the species of the genus Sergentomyia
Franca and Theodor from the Indian localities and
their possible role in kala-azar transmission
(Geevarghese et al., 2004), the correct identification
of the species becomes of significant value in the
study of epidemiology of leishmaniases and other
viral diseases.
Lewis (1967) and Artemiev (1978) while studying
Pakistani and Afghani sand flies respectively, did not
furnish morphometric measurements of mouth parts
(proboscis, mandible, hypopharynx, maxilla, buccal
cavity, pharyngeal armature), female genitalia and
male terminalia. Sketches of mandible, hypopharynx
and maxilla were also not furnished. Aslamkhan et
al. (1997, 1998) reported S. baghdadis from Lehri,
Sangsila and Dera Bugti areas of Balochistan
province but they neither described nor illustrated
morphology of taxonomic characters. Therefore, to
fill this gap, morphometric measurements and
photographs of diagnostic structures of S. baghdadis
as additional information, are being given here. A
comparative note of its taxonomic characters with its
closest allies is also given.
MATERIALS AND METHODS
During the routine entomological collection of
sand flies in Sindh & Punjab Provinces in 2006 the
author has collected several hundred Phlebotmine
sandflies including S. baghdadis and its closest
allies. The present investigation was carried out on
the materials (35 specimens of S. baghdadisi)
collected from Sindh and Punjab Provinces during
May, 2006 with sucking tubes and sticky traps. The
collected materials was preserved, processed and
dissected by conventional methods (Young and
Duncan, 1994). Identification of specimen was
carried out with the help of available literature
(Lewis, 1967, 1978; Artemiev, 1978). Morphometric
measurements and photographs were taken from
camera mounted Olympus microscope (BX41). Most
of the structures were measured with a low
magnification (X100) whereas spermatheca, ducts
and furca were examined under high magnification
Juma Khan Kakar
42
(X400). All given measurements are in mm.
Prepared permanent slides were deposited with the
author’s collection of sand flies, Department of
Zoology, University of Balochistan, Quetta.
RESULTS
The subgenus Parrotomyia Theodor includes
babu (Annandale), barraudi (Sinton), grekovi,
palestinensis, shortii (Adler and Theodor), montana
(Sinton), bailyi (Sinton) and the present one.
However, Kakarsulemnkhel (2004a, 2006) reported
babu, grekovi, and palestinensis from Balochistan
Province and examined them. The sub genus is
defined on the basis of following characters:
Comb-like buccal armature, lamp-glass shaped
pharynx, and elliptical capsular spermatheca, male
terminalia with 2 apical and two sub apical spines on
style or all spines apical.
1978). Mandible (X100) also blade like, 0.16-0.17
long, distal end of the mandible sharply pointed and
its inner margin carries small teeth, dental depth
0.015. Maxillary blade (X100) 0.16-0.17 long, stout
based, but gradually narrows towards its apex,
maxilla containing two rows of teeth, about 11 widely
spaced lateral teeth on outer edge near the apex
and its dental depth 0.035 and on inner edge after
some distance from the apex there is another row of
about 30 ventral teeth, its dental depth 0.08. Cibarial
cavity (Fig. 1A) (X100) 0.05 broad, 12-14
comparatively larger and pointed horizontal teeth
arranged in a concave line and 3-4 smaller median
teeth. Pigment patch of varying shape and size, also
present. Buccal plate shows a marked notch of
varying shape in the middle line. Posterior to the
buccal cavity, lies a large chitinous structure,
pharynx (X100) 0.13-0.15 long, anterior breadth
0.04-0.05, and 0.06-0.07 posterior breadth, several
toothed lines in the form of armature present at the
base of the pharynx.
Female genitalia (X400) (Fig. 1B)
Sergentomyia (Parrotomyia) baghdadis Adler
and Theodor, 1929
(Fig. 1A- 1B, 2A-2F)
Phlebotomus baghdadis Adler and Theodor, 1929,
Ann. Trop. Med. Parasit., 23: 281. Sinton, 1932.
Indian J. Med. Res., 20: 60; 1933.21: 422-423.
Sergentomyia (Parrotomyia) baghdadis Adler and
Theodor, Lewis, 1967, Bull. Brit. Mus. Nat. Hist.
(Ent.)19: 30, Artemiev, 1976. Medskaya Parazit.
45: 424; Artemiev, 1978. Ministry of Health,
Afghanistan, Kabul, 29. Lewis, 1978, Bull. Brit. Mus
Nat. Hist. (Ent.), 37: 258.
Female Wings, Palps, Antenna (X100)
Wings 1.3-1.35 long, breadth 0.3-0.33,
α / β=0.38, δ=0.05, gamma=0.25, П=0.13. Palps
(X100) 0.62-0.65 long, ratio 1,2.6,4,3.2,8.3, formula
1,2,3,4,5. Antennal segment (X100) A3 0.14-0.16
and A4 and A5 each 0.08-0.09. Ascoid formula 2
over A3-A15. Ascoids comparatively shorter (X400)
0.12 long and their positions (X400) on A3 0.71, on
A4 and A5 0.36. Papilla formula 1over A3-A5.
Positions of ascoids 0.79 and 0.75 respectively on
A3 and on A4 and A5 each.
Mouth parts, Cibarium, Pharynx (X100) (Fig. 1A)
Proboscis 0.2-0.21 long. Labrum hard sword
shaped structure, (X100) 0.15-0.16 long, relatively
narrow, sides parallel, apex bluntly pointed, margins
furnished with a series of long leaf like sensillae
closely together and numbering about 10 on either
side. Hypopharynx (X100) 0.15-0.16 long, also a
blade like structure, perforated by a narrow salivary
duct, apical end pointed, margins with short, thin and
soft serrations. Hypopharynx with soft serrations is a
character of the genus Sergentomyia (Artemiev,
Spermatheca (Fig.1B) 1.08-1.16 long, its head is
hard. 0.24 broad and plug-shaped while its base is
quite broader (0.64). Individual spermathecal duct
(X400) 1.2 long, arising from each spermatheca and
combine with one another to form a common duct
(1.1) finally falling in genital atrium (breadth 0.72).
Furca length 1.4.
Male Wings, Palps and Antenna (X100)
Wings 1.35-1.37 long, 0.3-0.32 broad, length
/breadth 4.28--4.5, α / β=0.46, δ=0.02, gamma=0.23,
П=0.09. Palpas (X100): 0.48-0.5 long, ratio
1,3.5,4,5,10 and formula 1,2,3,4,5. Antennal
segment 3 (X100) 0.15-0.16 long, A4 0.08 and A5
0.09 long, ascoid formula 1 over A3-A15, ascoid
(X400) very small in length (0.36 long), position of
ascoid on A3 0.65 and on A4 and A5 0.28.
Mouth parts, Cibarium, Pharynx (X100) (Fig. 2A)
Proboscis (X100) 0.17 long. Labrum: 0.10-0.11
long. Hypopharynx and maxilla each 0.1-0.11.
Mandible absent. Cibarium (X100) (Fig.2A): cibarial
cavity sharply angular, 0.03 long, below the ventral
plate a row 13-15 of minute and weak horizontal
teeth present arranged on a concave line, teeth
some times invisible. Pharynx (X100): 0.1-0.12 long,
anterior 0.03 and 0.05 posterior breadth, armature
very weak and confined at base of pharynx.
Male genitalia (X100) (Fig. 2B)
Coxite 0.2-0.22 long, 0.06-0.07 broad, style 0.1
long, 0.03 broad, a small ventral seta at 0.7 on style.
Paramere (Fig.2C) with blunt end, 0.12-0.13 long,
0.03 broad, narrow neck starts at 0.83 of length of
paramere, lower corners of paramere extending
downwards. Surstyle (Fig.2D) 0.16-0.17 long, apical
ends of paramere and surstyle almost at same level.
Taxonomic studies of Sergentomyia baghdadis (Adler and Theodor) in Sindh & Punjab
Filaments (F) with transverse striations. Sperm pump
(P) head (Fig.2E) like a semicircular funnel. F/ sperm
pump=2.5. Aedeagus sheath (Fig.2F) short, 0.070.08 long, apical end of aedeagus sheath rounded,
colorless but slightly curved ventrally.
Material examined
18 ♀, 15 ♂, Sindh, Punjab, May, 2006.
Distribution
Iraq, Iran, southern Afghanistan, Pakistan.
Present study, new record: Sindh: Dadu,
Jacobabad, Khairpur Nathan Shah, Mehar, Multan,
Nau Shehro Feroz, Qambar Ali Khan, Sehwan
Shareef, Sukkur. Punjab: Dera Ghazi Khan, These
are endemic foci of cutaneous leishmaniasis (CL).
British Museum (Natural History): Dera Ismail Khan,
Jhelum, Kandhkot, Lahore, Lyallpur, Pano Aqil,
Peshawar, Sargodha, Tank. Landi Kotal, Mir
Mohammad, Rawalpindi, Said Pur, Taxilla (Lewis,
1967).
Comparative note
The babu group (babu and baghdadis) is entirely
isolated with apomorphies of presence of hind notch
in the ventral plate of cibarium which is of particular
importance because both species have better
developed characters. Their oblong spermatheca
with broader distal end isolates them among its
subgroups. They also share synapomorphies of hind
notch at ventral plate of cibarium. The two species,
however, are clearly separated on the bases of
buccal teeth. S. baghdadis appears unique in this
group with aut-apomorphy of presence of smaller
median teeth in the cibarium. The grekovi and
palestinensis-group maintain their separate identity
but having synapomorphies of buccal teeth arranged
on a straight line, hind notch at ventral plate lost,
presence of large triangular pigment patch, almost
spherical spermathecae and smooth genital
filaments. S. grookovi can be easily differentiated for
its autapomorphies of short genital filaments and
pharyngeal armature directed obliquely down. The
palestinensis can be discriminated by its
autapomorphic characters of about 15-18 parallel,
arrow like buccal teeth, each with nodular thickening
near its center and also with a second row of about
19-21 punctiform denticles at the base of the longer
ones and with pharynx much dilated posteriorly.
However, S. baghdadis shares some of its
characters such as buccal cavity with 20 hardly
visible teeth arranged in concave lines, pigment
patch very pale or absent, genital filaments with
transverse striations in male, comb-like cibarial
armature, lamp-glass shaped pharynx, and long
spermathecal capsule, ventral plate of cibarium with
angular deep notch at center. It is distinct, however,
in its group in having 15-18 buccal teeth including 34 very small median teeth and weak pharyngeal
43
armature, comparatively longer spermatheca with
larger head and basal part of capsule much broader
is closely related to S. babu (Annandale) in which
male cibarium with about 20 hardly visible uniform
teeth arranged in concave line, there is additional
row of smaller denticles at their bases, pigment
patch very small or absent, paramere with slightly
beaked end and shorter than the length of surstyle,
lower corners of paramere extending downwards, in
female ventral plate of cibarium with angular deep
notch and about 28 sharply pointed uniform
horizontal teeth arranged on a concave row but S.
baghdadis can easily be separated from the same in
having 12-14 lateral cibarial teeth and 3-5 very small
in the center, pharyngeal armature weak,
comparatively longer spermathecal capsule with
larger head and basal part of capsule much broader.
DISCUSSION
Results of the present study were compared with
the published data of S. baghdadis (Adler and
Theodor) from other territories (Table 1). Length /
breadth ratio of the wings of our specimens (♀= 4.04.3; ♂=4.2-4.5) were observed to be shorter than of
the results (♀= 4.5-4.7; ♂=4.7-4.9) reported by Lewis
(1967). Similarly, A3 of present samples were found
a little smaller (♀= 0.14-0.15; ♂=0.15-0.16) than of
the findings (♀= 0.15-0.17; ♂=0.14-0.19) given by
Lewis (1967). Number of cibarial teeth in Pakistani
flies whether in present study or in the past study
conducted by Lewis (1967), were almost same and
less in number (about 18) however, more cibarial
teeth (20-25) were reported by Artemiev (1978) in
Afghanistan S. baghdadis. F/ sperm pump ratio in
Pakistani ♂ S. baghdadis were observed almost
similar whether in present study or in past study of
Lewis (1967).
Though minor variations in morphometric
measurements of taxonomic characters were found
when were compared with published data from other
territories, however, present work is in conformity
with the findings of Lewis (1967) and Artemiev
(1978). The effect of different ecological factors like
temperature, relative humidity ecological niche on
the growth and size of structures of flies can not be
ruled out. Belazzoug et al.(1982) while working in
different ecological zones of Algeria has shown that
number of cibarial teeth varies according to certain
climatic factors (mainly humidity).
It is hoped that present findings would provide
the basis for further research on sand flies taxonomy
in the country and indirectly also on other aspects
that are essential for the control of sand flies and the
disease leishmaniases. Keeping in view of its
presence in human residences in the areas of C.L. in
Sindh and Punjab Provinces, vectorial role of S.
baghdadis needs to be further investigated.
44
Juma Khan Kakar
Fig.1. Female Sergentomyia (Parrotomyia) Baghdadis (Adler and
Theodor): A, Cibarium (X400); B, Spermatheca (X400).
Fig.2. Male Sergentomyia (Parrotomyia) baghdadis (Adler and Theodor):
A, Cibarium (X400); B, Male Genitalia (X200); C, Paramere (X200); D,
Surstyle (X200); E, head of spermathecal pump (X400); F, Paramere
(X200) and Aedeagus (X400).
Taxonomic studies of Sergentomyia baghdadis (Adler and Theodor) in Sindh & Punjab
Table-1. Comparison of Taxonomic Characters (mm) of S. baghdadis
(Adler & Theodor)
Characters
Sindh
Sand flies
Present Study
Sinton
(1932)
Lewis (1967)
Afghanistan
(Artemiev,
1978)
♀
♂
1.30 – 1.35
1.35 – 1.37
-
1.51- 1.72
1.39 – 1.64
-
♀
♂
4. 0 – 4. 3
4. 2 – 4.5
-
4. 5 – 4. 7
4. 5 – 4. 9
-
♀
♂
0. 38
0. 46
-
0. 3 – 0.8
-
-
♀
♂
0. 14 – 0. 15
0. 15 – 0. 16
-
0. 15 – 0. 17
0. 14 – 0. 19
-
♀
♂
0. 93 – 1. 0
1. 45 – 1. 50
-
1.0 – 1. 1
1. 0 – 1. 2
-
♀
♀
11
30
-
-
-
♀
12-14 larger +
3-4 smaller,
median
13-15 hardly
visible
16-18
7 large, each side +
4 small
in centre,
16-20 +4-5
central,
Scarcely visible
14- 16
2. 5
2. 5
Beaked,
Lower corners
extending
downwards
Apical end
rounded, but
slightly curved
ventrally
colorless
-
-
-
-
Wing Length
Wing
L / Breadth
Alar Index
A3 Length
A3 / Labrum
Maxilla Teeth
Lateral
Ventral
Cibarial Teeth
♂
Filament /Pump
Paramere
♂
♂
Adeagus
♂
Beaked,
Lower corners
extending
downwards
Apical end
rounded, but slightly
curved
ventrally
colorless
45
Juma Khan Kakar
46
REFERENCES
ARTEMIEV, M.M. (1978). Sand flies (Diptera,
Psychodidae, Phlebotominae) of Afghanistan. iv+87 pp. Kabul.
ASLAMKHAN,
K.,
ASLAMKHAN,
M.
AND
AZIZULLAH. (1997). The distribution records of
sand flies (Phlebotominae) of Pakistan and
Kashmir from 1908 to 1996. Pakistan J. Zool.,
29: 351-360.
ASLAMKHAN, K., ASLAMKHAN, M. AND AZIZULLAH.
(1998). Biodiversity of sand flies of Pakistan and
Kashmir. Pakistan J. Zool., 30: 13-21.
BELAZZOUG, S., MAZHOUL, D., ADDADI, K.
AND DEDET, J.-P. (1982). Sergentomyia minuta
parroti (Adler and Theodor, 1927) en Algierie
(Diptera: Psychodidae). Ann. Parasit. Hum.
Comp., 57: 621-630.
GEEVARGHESE, G., AMAKALLE, V.A., JADI, R.,
KANOJIA, P.C. JOSHI, M. AND MISHRA, A.C.
(2004). Detection of Chandipura Virus from
Sand Flies in the Genus Sergentomyia (Diptera:
Phlebotomidae) at Karimnagar, District, Andhra
Pradesh, India. J. Med. Entomol. 10: 495-496.
KAKARSULEMANKHEL, J.K. (2004). Composition of the
Phlebotominae Fauna (Diptera:Psychodidae) in
Balochistan, Pakistan. J. Biol. Sci., 4 (3): 391-392.
KAKARSULEMANKHEL, J.K. (2006). Sand flies of
Pakistan (Diptera: Psychodidae), pp.87-98. In:Role of
Insect Taxonomy, Systematic in sustainable
Agriculture. Proc. Natl. Workshop.PARC/NARC. Feb.
13-14, 2006, Islamabad.
LEWIS, D.J. (1967). The phlebotomine sand flies of
west Pakistan (Diptera: Psychodidae). Bull.Brit.
Mus. Nat. Hist. (Ent.), 19: 1-57.
LEWIS, D.J. (1978).The phlebotomine sand flies
(Diptera, Psychodidae) of the Oriental region.
Bull. Brit. Mus. Nat. Hist. (Ent.): 37: 217-343.
THEODOR, O. (1948). Classification of the Old
World species of the sub family Phlebotominae.
Bull. Ent. Res., 39: 85-111.
YOUNG, D.G. AND DUNCAN, M.A. (1994). Guide to
the Identification and Geographic distribution of
Lutzomyia sand flies in Mexico, the West Indies,
Central and South America (Diptera: Psychodidae).
Mem. Am. Entomol. Inst., 54: 1- 881.
POTENTIAL OF SWEET FLAG RHIZOME OIL AND CUSTARD APPLE
SEED OIL AGAINST THE MAJOR SUCKING PESTS OF COTTON, AS
COMPARED WITH CONFIDOR + DELTAPHOS, AT ARI-TANDOJAM,
SINDH-PAKISTAN
TARIQ*, R.M., NAQVI**, S.N.H., ZAFAR*, S.M.N. AND BURRERO***, A.S.
*Department of Zoology, University of Karachi, Karachi-75270, Pakistan.
**Department of Pharmacology, Baqai Medical University, Super Highway,
Toll Plaza, Karachi-Pakistan.
***Entomology Section, Agriculture Research Institute, Tandojam, Sindh-Pakistan.
ABSTRACT
The two local plants, Acorus calamus (Sweet flag/Batch) rhizome oil and Annona squamosa (Custard
apple/Seeta phall/Shareefa) seeds oil was tested against three major sucking pests, whitefly (nymph & Adult)
(Bemisia tabaci), Thrips (Thrip tabaci) and Jassids (Amrasca devastant). The results of these were compared
with that of Confidor + Deltaphos (CD) used as standard. The controlling power (potential) of the two candidate
tested plants and standard was 65.02, 63.23 and 78.02%, respectively by A. calamus (AC) and A. squamosa
(AS) and standard (CD) after two weeks, against Adults of whitefly. Whereas 70.00, 66.86 and 80.47% against
whitefly nymph, in the same sequence. While the potentiality of AC, AS and CD, against Thrips was, 46.93,
46.61 and 69.92%, respectively after two weeks. Whereas against Jassids the efficacy of AC, AS and CD was
found to be 60.22, 62.43 and 82.32%, respectively after two weeks. Overall the phytopesticides AC and AS were
found having less effective than Confidor + Deltaphos, but even that the candidate oils gave remarkable good
results. More research, struggle and results are needed to prove them as pesticides.
Key words: Acorus calamus, Annona squamosa, Confidor + Deltaphos, Major sucking pests, Cotton.
INTRODUCTION
Phytopesticides are eco-friendly and safe from
the pollution point of view. Whereas the commercial
and synthetic pesticides such as pyrethroids are
reported hazardous because their poisoning
syndromes, synergies and therapy has been
reported by Ray and Philip (2000). The pyrethroid
induced paresthesia, a central or local toxic effect
have been reported by Wilks (2000). Therefore, to
chalk out new alternates of commercial and synthetic
pesticides the researchers are busy in testing
different plants extracts, such as the neem which is
providing a good alternate as a phytopesticide and
several formulations are being used in different parts
of the developed and developing countries. Several
plants and trees are tested such as marrango tree by
Ermel et al. (1991), Cameroon by Kouninki et al.
(2005), Cerrado Plant extract by Rodngues et al.
(2006), Pamp and Occimum by Verma et al. (2006),
but more attention have been given to Acorus
calamus (AC) and Annona squamosa (AS), such as
Ahmed et al. (2000), Alesso et al. (2003), Johri et al.
(2004), Morio and Kuriyama (2005), Miguel et al.
(2006). Naqvi, et. al. (2006), Akbar et al. (2007),
Tariq et al. (2007). Therefore keeping in mind this
strategy we have tested the oil formulation from
Acorus and Annona against the pests of cotton crop
at field level in agriculture sector.
MATERIALS AND METHODS
The extracted oil from the dried rhizome of
Acorus calamus was coded as “AC” whereas the oil
of Annona squamosa, coded as AS was extracted
from the seeds. Both oil formulations contained 2%
emulsifier (Twin-80) and after preparing layout the
candidate pesticides were sprayed on cotton crop
after taking pre-treatment observation in all the
cases. Other precautions were also taken in
agriculture treatments, such as the humidity, the
wind, the rain, the temperature etc. The observations
were recorded after 24, 72 hours, and one week. In
the case of whitefly nymph only the observations
were recorded after 72 hours and one week. The
results were analyzed on standard pattern and
compared with standards Confidor + Deltaphos. The
experiments were conducted on cotton crop at
Integrated Pest Management (IPM) Laboratory,
Agriculture Research Institute (ARI) Tandojam, in the
year 2001. The Randomized Complete Block Design
(RCBD) method was adopted. Six replications of 4
treatments was done and the average value was
used in the analyses.
RESULTS
Four treatments were designed T1 (AC), T2
(AS), T3 (Confidor + Deltaphos) and T4 (Control).
After taking pre-treatment observation, the designed
plots for AC, AS & C+D were sprayed at the dose of
1250ml/acre of AC & AS, whereas the doe of C+D
was 800ml/acre in (1:1) ratio. While the control plot
was not sprayed, the experiments were set at six
places as replicates at a time, on a cotton crop
shown on 31-03-2001. The experiments were done
in July 2001.
All the plots designed for experiments of AC,
AS, C+D, were sprayed on same day after pre-
Tariq et al.
48
treatment observation. The observations were taken
after 24, 48, 72 hours, one week and two weeks.
The reduction percentage after 24, 48, 72 hours, one
week and two weeks each by AC, AS and C+D
against whitefly adults per leaf has been shown in
Table-1. Whereas against whitefly nymphs per leaf
has been shown in Table-2.
The pre-treatment observation and observation
after spray of 24, 48, 72 hours, one week and two
weeks, with reduction percentage of the above
duration against Thrips/leave have been shown in
Table-3, whereas against Jassids/leaf has been
shown in Table-4. It may be noted in all four case i.e.
whitefly adults, while fly nymphs, Thrips and Jassids
that the efficacy of AC & AS is less as compared to
Confidor + Deltaphos (C+D), but even that they (AC
& AS) are remarkably comparable with Confidor +
Deltaphos after 2 week. It means that the
phytopesticide may give control of insects upto 15
days (2 weeks) effectively which is less than C+D,
but is more safer and beneficial as compared these
synthetic and conventional pesticides.
Table-1. Pre-treatment observation & average reduction percentage of whitefly adults/leaf.
No. of
Treatments
T1 Acorus
calamus
T2 Annona
squamosa
T3 Confidor +
Deltaphos
T4 Control*
PreTreatments
1.25
24 Hrs.
48 Hrs.
72 Hrs.
One week
Two week
0.63
0.81
0.96
1.86
0.78
1.29
0.62
0.74
1.01
1.19
0.82
1.16
0.40
0.31
0.40
0.54
0.49
1.65
2.15
2.68
2.73
2.23
24 Hrs.
70.69
71.16
81.39
48 Hrs.
67.77
72.38
88.43
2.71
Reduction %
72 Hrs.
64.57
62.73
85.24
One week
31.86
56.41
80.22
Two week
65.02
63.23
78.02
Table-2. Pre-treatment observation & average reduction percentage of whitefly nymphs/leaf.
No. of
Treatments
T1 Acorus
calamus
T2 Annona
squamosa
T3 Confidor +
Deltaphos
T4 Control*
PreTreatments
0.93
24 Hrs.
0.69
0.62
0.80
0.67
0.49
1.00
0.95
0.53
0.74
0.63
0.56
0.87
0.33
0.27
0.38
0.27
0.33
1.38
1.96
2.26
2.43
Reduction %
72 Hrs.
67.08
69.54
84.36
2.07
1.69
24 Hrs.
64.79
51.53
83.16
48 Hrs.
48 Hrs.
72.56
76.54
88.05
72 Hrs.
One week
One week
67.63
69.56
86.95
Two week
Two week
70.00
66.86
80.47
Table-3. Pre-treatment observation & average reduction percentage of Thrips /leaf.
No. of
Treatments
T1 Acorus
calamus
T2 Annona
squamosa
T3 Confidor +
Deltaphos
T4 Control*
PreTreatments
4.46
24 Hrs.
1.63
1.77
1.73
3.22
3.37
4.34
1.49
1.42
1.62
3.71
3.39
4.01
1.05
0.75
0.70
1.73
1.91
4.27
4.27
4.14
3.95
Reduction %
72 Hrs.
56.20
58.98
82.27
7.22
6.35
24 Hrs.
61.82
65.10
75.41
48 Hrs.
48 Hrs.
57.24
65.70
81.88
72 Hrs.
One week
One week
55.40
48.61
76.04
Two week
Two week
46.93
46.61
69.92
Potential of sweet flag rhizome oil and custard apple seed oil against the major sucking pests
49
Table-4. Pre-treatment observation & average reduction percentage of Jassids/leaf.
No. of
Treatments
T1 Acorus
calamus
T2 Annona
squamosa
T3 Confidor +
Deltaphos
T4 Control*
PreTreatments
1.30
24 Hrs.
0.62
0.78
0.66
0.79
0.72
0.99
0.64
0.59
0.48
0.59
0.68
1.05
0.24
0.26
0.23
0.34
0.32
1.51
2.23
2.25
2.43
Reduction %
72 Hrs.
72.84
80.24
90.53
2.44
1.81
24 Hrs.
73.27
72.41
89.65
48 Hrs.
48 Hrs.
65.33
73.77
88.44
72 Hrs.
One week
One week
67.62
70.82
86.06
Two week
Two week
60.22
62.43
82.32
*The reduction% in control was nil, due to which it has not been shown in reduction columns.
DISCUSSION
Johri et al. (2004) reported comparative toxicity
of seven indigenous botanical extracts against the
infestative stage of three insect pests of agricultural
importance. The plants used by them were
Tephrosia vogelii Hook F. (leaves & seeds –
rotenoids), Annona squamosa Linn (seed-oil),
Tribulus terrestris Linn (leaves-saponins), Pongamia
glabra Vent (leaves & seed-oils) were all above
extracted in petroleum ether, and ethyl alcohol
extract of Thevetia nerifolia Juss (fresh fruit-Thevetin
glycoside) and Duranta repens Linn (fruit-alkaloids)
while both the solvents were applied in two steps for
Caesalpinia crista Linn. seed-glycoside and fatty oil)
were screened for their toxicity against three
phytophagous insect pests, viz. Painted bug
Bagrada cruciferarum Kirk on mustard and cabbage,
white butterfly Pieris brassicae Linn. on cabbage and
blister beetle, Mylabris pustulata Thunb on mung.
The T. nerifolia showed higher toxicity followed by A.
squamosa, T. vogelii and C. crista.
In the present work Acorus calamus (AC) &
Annona squamosa (AS) were used to control
whitefly (nymphs + Adults), Thrips & Jassids on
cotton crop. Both AC & AS gave more than 60%
control in the case of whitefly (N+A), whereas
standard gave more than 70% control after two
weeks. Whereas in the case of Thrips AC, AS &
C+D gave 46.93, 46.61 & 69.92% control
respectively. While in the case of Jassids AC, AS &
C+D gave 60.22, 62.43 & 82.32% control
respectively after two weeks.
Shuijin et al. (2006) reported the toxicity more
than of 10 insecticides by two methods, the topical
application and the leaf dipping method. The aim of
this study was to monitor the resistance against
these insecticides in common cutworm. Spodoptera
litura (Fabricius). The topical application method was
found more sensitive for monitoring the resistance to
contact insecticides whereas the leaf dipping method
was more sensitive for monitoring resistance to
insecticides with stronger stomach action. The
results were conducted by using susceptible strain
and were compared with two field populations of S.
litura. The two field populations were shown to have
high
resistance
to
organophosphates
and
carbamates. However, the Nanjing population had
low resistance to profenofos and methomyl. No
resistance to monosultap, abamectin, hexaflumuron
and fipronil was found in these populations. They
proposed to avoid the use of pyrethroids
organophosphates and the carbamates should be
limited and the other insecticides with no resistance
should be used rationally and in rotation to delay
resistance development. Whereas in the present
work keeping in mind the above resistance problem
against
the
pyrethroids,
organophosphates,
organochlorinates, and carbamates, the botanical
(plant origin)/phytopesticides are used against
sucking pest of cotton crop in field, i.e. Ac & AS.
Naqvi et al. (2006) reported the efficacy of
Annona oils as biopesticide against the major
sucking pests (whitefly nymphs + Adults, Jassids &
Thrips) of cotton on field level at Central Cotton
Research Institute (CCRI) Multan (Punjab-Pakistan).
The results were compared with Mospilan &
Tamaron. The results were based on 24, 72 hours &
one week observation. They used 1500ml/acre dose.
But in the present work 1250ml/acre dose was used
for the experiments against the above major sucking
pests of cotton i.e. whitefly nymphs + Adults, Jassids
& Thrips. They sued 125ml/acre of Mospilan against
whitefly (N+A) and 500ml/acre of Tamaron against
Thrips & Jassids. They concluded that AC & AS
proved less effective as compared to Mospilan &
Tamaron but may be successfully used against
these pests. The same conclusion was drawn in the
present work after the experimentation at ARITandojam.
Akbar et al. (2007) reported the efficacy of
Biosal a neem product of HEJ, Karachi-Pakistan
against Amrasca devastans, the Jassids on brinjal
crop. They concluded that the Biosal being a neem
based formulation showed less toxic effects as
compared to Endosulfan & Profenophos, may be
due to strong anti-feedant, insect growth regulatory
Tariq et al.
50
(IGR) effect of neem. In the present work, as well,
the Calamus & Squamosa oil proved less effective
against whitefly (N+A), Thrips & Jassids as
compared to confidor + Deltaphos, preferably due to
the same above reasons.
Tariq et al. (2007) reported the toxic effects of A.
calamus & A. squamosa against the bollworms
(American, spotted & pink bollworm) on cotton crop.
They reported 45.76, 57.20, 48.30, 58.05 & 47.03%
control of American bollworm by AC, AS, Karate,
Polytrin-C & Curacron respectively, upto two weeks.
Whereas against spotted bollworm, the control after
two weeks by AC, AS & Confidor + Deltaphos was
41.12, 58.15 & 78.66% respectively. While the
control after two weeks in the cast of pink bollworm
by AC, AS & C+D was 51.54%, 58.17 & 81.94%
respectively. But in the present work the experiments
were done against the major sucking pests of cotton
by Acorus calamus (AC) & Annona squamosa (AS)
to control whitefly (nymphs + Adults), Thrips &
Jassids on cotton crop. Both AC & AS gave more
than 60% control in the case of whitefly (N+A),
whereas standard gave more than 70% control after
two weeks. Whereas in the case of Thrips AC, AS &
C+D gave 46.93, 46.61 & 69.92% control
respectively. While in the case of Jassids AC, AS &
C+D gave 60.22, 62.43 & 82.32% control
respectively after two weeks.
REFERENCES
AHMED, I., AHSAN, T., TABASSUM, R., AZMI, A.
AND NAQVI, S.N.H. (2000). Effects of Acorus
calamus extract and cypermethrin on enzymatic
activities in Sitophilus oryzae. J. Exp. Zool.
India. Vol. 3(2): 169-173.
AKBAR, M.F., YASMIN, N., NAQVI, S.N.H., KHAN,
M.F. & NAZ, F. (2007). Relative efficacy of
biopesticide in comparison with conventional
pesticides against Amrasca devastans Dist. on
Brinjal crop. Pakistan j. entomol. Karachi, 22 (1&2):
1-3.
ALESSO, E., TORVISO, R., LANTANO, B., ELRICH,
M.,
LILIANA,
M.,
FINKIELSZTEIN,
MOLTRASIO, G., AGUIRRE, J.M. AND
BRUNET, E. (2003). Synthesis of 1-ethyl-2methyl-3-arylindanes. Stereochemistry of finemembered ring formation (online) ARKIVOC
2003 (X): 283-297.
ERMEL, K., KALINOWSKI, H.O. AND SCHMUTTERER,
H. (1991). Isolation and characterization of narrangin,
a new insect growth regulating (IGR) substance from
the seed kernels of the marrango tree, Azadirachta
excelsa (Jack). J. Appl. Entomol. 112(5): 512-519.
JOHRI, P.K. MAURYA, R., SINGH, D., TIWARI, D.
AND JOHRI, R. (2004). Comparative toxicity of
seven indigenous botanical extracts against the
infestative stage of three insect pests of
agricultural importance. Journal of Applied
Zoological Researches. 15(2): 202-204.
KOUNINKI, H., HAUBRUGE, E., NOUDJOU, F.E.,
LOGNAY, G., MALAISSE, F., NGASSOUM,
M.B., GOUDOUM, A., MAPONGMETSEM,
P.M., NGAMO, L.S.T. AND HANCE, T. (2005).
Potantial use of essential oils from Cameroon
applied as fumigant or contact insecticides
against Sitophilus zeamais Motsch (Coleoptera:
Curculionidae). Communications in Agricultural
and Applied Biological Sciences. 70(4): 787-792.
MIGUEL, B., ZAVALA, F., SISNIEGAS, M.,
ZAVALETA, G., MOSTACERO, J. AND
TARAMONA, L. (2006). Larvicidal evaluation of
aqueous suspensions of Annona muricana
Linnaeus “custard apple” against Aedes aegypti
Linnaeus
(Diptera,
Culicidae).
REVISTA
PERUANA DE BIOLOGIA 12(1): 145-152.
MORIO, T. AND KURIYAMA, M. 2005. Pests of
Annona cherimola (Annonaceae) in a newly
introduced area, with respect to their host range.
Journal of the Entomological Research Society.
7 (Part 3): 1-12.
NAQVI, S.N.H., TARIQ, R.M., ZAFAR, S.M.N. &
ATTIQUE, M.R. (2006). Efficacy of Acorus calamus
(AC) rhizome oil and Annona squamosa (AS) seed
oil against sucking pests of cotton at CCRI-Multan
(Punjab) as compared to Mospilan & Tamaron.
Pakistan j. entomol. Karachi, 21 (1&2): 23-27.
RAY, D.E. AND PHILIP, J.F. (2000). Pyrethroid
insecticides: Poisoning syndromes, synergies
and therapy. Journal of Toxicology Clinical
Toxicology. 38(2): 95-101.
RODNGUES, A.M.S., PAULA, J.E.D., DEGALHER, N.,
MOLEZ, J.F. AND ESPINDOLA, L.S. (2006).
Larvicidal activity of some Cerrado plant extracts
against Aedes aegypti. Journal of the American
Mosquito Control Association. 22(2): 314-317.
SHUIJIN, H., XU, J. AND HAN, Z. (2006). Baseline
toxicity data of insecticides against the common
cutworm Spodoptera litura (Fabricius) and a
comparison of resistance monitoring methods.
International Journal of Pest Management 52
(3): 209-213.
TARIQ, R.M., NAQVI, S.N.H., ZAFAR, S.M.N. &
BURRERO, S. (2007). Toxic effects of botanical
pesticide, from Acorus calamus (AC) and Annona
squamosa (AS) against bollworms at ARITandojam Sindh-Pakistan. Pakistan j. entomol.
Karachi, 22 (1&2): 31-36.
VERMA, P.R., SUBBURAJU, T. AND BALAKRISHNAN, N.
(2006). Larvicidal activity of Artemisia nilagirica
(Clarke) Pamp. and Ocimum sanctum Linn. – A
preliminary study. JOURNAL OF NATURAL
REMEDIES. 6(2): 157-161.
WILKS, M.F. (2000). Pyrethroid induced paresthesia: A
central or local toxic effect? Journal of Toxicology
Clinical Toxicology. 38(2): 103-105.
STUDIES ON SAMPLING TECHNIQUES TO MONITOR ADULT
POPULATION OF WHITEFLY, BEMISIA TABACI (GENN.)
IN CUCURBIT FIELD
ABDUL GHANI LANJAR*, MUHAMMED KHAN LOHAR*, HAKIM ALI SAHITO*,
NAHEED BALOCH** AND ASHFAQUE AHMED NAHIYOON*
*Department of Entomology, Sindh Agriculture University, Tandojam
**Department of Zoology, University of Sindh, Jamshoro
ABSTRACT
A half - acre field of melon, Cucumis melo L. was selected at Tandojam during spring, 2000 for
sampling adult population of whitefly, Bemisia tabaci (Genn.). The methods of sampling were direct and
indirect count. In the direct count, 30 leaves were randomly selected and examined from 2nd, 3rd and
5th nodes of the creepers. In the indirect count, yellow sticky traps of different shapes, i.e., flat,
cylindrical and round were used. The traps were mounted at 6", 12", and 18" above ground. Highest
population of B. tabaci 6.56 ± 1.34 per leaf was recorded on the 2nd- node leaves as compared to 3rd,
2.62 ± 0.52 and 5th-node leave 2.52 ± 0.56. More activity 6.56 ± 1.34 per leaf was recorded at 8.00
a.m. than 4.03 ± 0.83 at 12.00 noon. Highest catches 22.87 ± 2.69 per trap were recorded on cylindrical
and lowest 9.24 ± 1.39 on round traps. Traps mounted at 6" above ground level gave 14.37 ± 2.02
catches of B. tabaci adults. Population counts in all three techniques were highly significant (P<0.01).
Key words: Bemisia tabaci, Melon, Population and Sampling.
INTRODUCTION
placed parallel to plant height catch more whiteflies
(Malamed-Madjar et al. 1982).
The whitefly, Bemisia tabaci (Gennadius) is one
of the most important members of Family
Aleyrodidae for its grave ravages on tropical and
subtropical agriculture. It is a polyphagus insect pest
recorded on more than 506 species of plants and
field crops (Mound and Halsey, 1978). Greathead
(1986) updated this information, who listed the host
range up to 540 species belonging to 77 plant
Families. In Pakistan, it was recorded on 160 plant
species belonging to 113 genera of 42 families
including crops, ornamentals, fruit and forest trees,
and weeds (Attique et at., 2003; Lanjar and Sahito,
2005).
In a cotton field, the sample unit per plant
consisted of first two fully expanded main terminal
leaves (Musuna, 1986). The sampled leaves with high
and low infestations of B. tabaci were found between
8th leaves from top to bottom (Fowler, 1956; Mound,
1965). Melamed-Madjar et al. (1982) reported that the
most infested leaves were between 5th and 6th nodes.
Gusmao et al. (2005) mentioned the most
appropriate sampling methods were beating one
apical leaf in the tray to assess adults and by direct
counting of nymphs on a basal leaf of tomato plants.
Sequeira and Naranjo (2008) recommended that
minimum sample of 20 leaves at nodes 3, 4 or 5
below the terminal as the most parsimonious and
practical sampling protocol for cotton fields. B.
tabaci count on plant leaves was considerably high
early in the morning, because the adults were less
mobile (Gerling and Horowitz, 1984). The work of the
previous authors demonstrate that satisfactory
sampling method to manage B. tabaci on cucurbit
crops is utmost important.
Proper sampling is essential for estimating the
number of insect pests in a field crop. Various
Sampling methods have been used to monitor B.
tabaci population. The immature stages of whitefly
are hard to detect (Ohnesorge and Rapp, 1986).
The adult dispersed in field for a considerable
distance (Byrne, 1999). They are characteristically
attracted to yellow surfaces (Cohen, 1982). The use
yellow sticky trap is a major tool in monitoring adult
population (Berlinger, 1980; Gerling and Horowitz,
1984; Butler et al. 1985). Various shapes and sizes
of the traps such as flat and cylindrical have been
satisfactorily used to monitor B. tabaci population
(Byrne et al. 1986; Youngman et al. 1986). Traps
Observations
The investigations on the sampling techniques for
monitoring adult population of B. tabaci were
conducted in a melon field at Tandojam during spring
Lanjar, A.G. et al.
52
season. Melon crop was sown on an area of 1/2 acre.
The whitefly adult populations on melon were
sampled by using direct and indirect counts.
Indirect Count
Yellow traps of different shapes namely flat,
cylindrical and round were placed, two near the sides
and one trap in center of the melon field. Three
cylindrical shaped traps were placed above ground at
different heights of 6", 12" and 18". The traps were
coated with adhesive grease.
Observations on
population count were taken twice a week. After each
observation the grease on yellow traps was changed.
There were three (trap sizes and heights) replications
of each sampling technique.
Trap Size
Flat trap:
Made of a plastic sheet (12" x
12") fitted into one meter long
wooden stick.
Cylindrical trap:
Made of a plastic pipe of 12"
length and 6" diameter filled with
plaster of paris to support onemeter long wooden stick.
Round trap:
Made of a glass globe of 8"
diameter filled with plaster of
Paris to support one -meter long
wooden stick.
4th week of April, where 42.33±3.56 adult B. tabaci
were recorded on cylindrical trap, 26.33±4.00 on flat
and 18.66±3.13 on round trap. The average adult B.
tabaci count on cylindrical trap was 22.87±2.68, on
flat trap was 14.37±2.02 and on round trap was
9.24±1.39.
Results show that the use of cylindrical traps is
better than flat and round traps. B. tabaci adults were
trapped from all directions on the cylindrical traps in
the melon crop. This indicates that B. tabaci
population was equally distributed in the melon field.
The results showed that population of adult B.
tabaci caught on three traps were highly significant.
The present results are in agreement with the findings
of earlier authors. Berlinger (1980) reported that B.
tabaci was strongly attracted to yellow color surface.
Butler et al. (1985) stated that yellow sticky traps
became a major tool in monitoring of B. tabaci
population. Flat traps of various sizes have generally
been used (Butler et al. 1986). However, cylindrical
types have given better results (Byrne et al. 1986,
Youngman et al. 1986).
30
Direct Count
Population densities of adult B. tabaci were
estimated by direct count on melon leaves. The
sampling unit per leaf comprised of fully expanded
leaves of 2nd, 3rd and 5th nodes of melon creepers.
Thirty leaves of different creepers were randomly
selected and examined twice a week for B. tabaci
population. The counting of B. tabaci was carried out
between 8.00 a.m. and 12.00 noon.
25
20
15
10
5
0
Fl at
cy l i ndr i cal
r ound
Fig.1. Mean catches per trap of Bemisia tabaci on
various types of yellow sticky traps mounted in melon
field during spring season.
Whitefly Activity at Different Ground Levels
Indirect Count
Population of B. tabaci was estimated on melon
from germination until harvest of the crop. Figure -1
shows that at initial growth stage of the creepers the
number of adults caught in the traps was very low.
The first adult catches on the different traps were
recorded during 1st week of March just after crop
germination. During initial catches 5.00±0.94,
2.66±0.72 and 1.00±0.47 adults per trap were
recorded on cylindrical, flat and round traps,
respectively.
Population of B. tabaci fluctuated later on and
showed two peaks. The first population peak was
observed during 4th week of March, where the adults
caught on cylindrical, flat and round traps were
50.00±4.11, 38.00±4.32 and 26.33±2.59 per trap,
respectively. The second peak was recorded during
Figure 2 shows that the population of B. tabaci
varied at the different ground levels. At initial growth
stage, activity of B. tabaci was higher at 6" above
ground level than at12" and 18". The activity of B.
tabaci depended proportionately on the growth of
creeper. Two peaks namely, the 1st during 4th week
of March, and the 2nd during 4th week of April were
observed. At 1st peak the count of adult B. tabaci
was 38.00±4.32, 25.00±3.09, and 7.00±0.94
adults/trap at 6", 12" and 18" respectively. At the
second peak the count of adult B. tabaci was
26.33±4.00, 17.33±4.83 and 13.33±2.88 per trap
respectively. The mean total count of adult B. tabaci
at 6", 12" and 18" was 14.37±2.02, 7.81±1.43, and
3.61±0.68, respectively.
The result of this experiment indicates that in
melon field the flight and dispersal activities of B.
tabaci adults were maximum at 6" above ground level,
and lower at 18". It clearly shows that B. tabaci retain
Studies on sampling techniques to monitor adult population of whitefly, Bemisia tabaci (Genn.)
53
their activities mostly up to the height of melon
creepers. The present results are in agreement with
the findings of earlier researchers. Sharaf (1982)
reported that horizontally placed traps gave high
catches per trap than those placed vertically. Traps
fixed at ground level gave higher catches in fallow
than in a cotton field (Gerling and Horowitz, 1984).
Melamed Madjar et al. (1984) reported that whitefly
catches within field and 4 meter away from border
were the same, when the traps were placed at plant
height.
Suitable Timing of B. tabaci Sampling
Direct Count
In the direct count, numbers of B. tabaci on 2nd,
3rd and 5th node on leaves were recorded (Figure
3). In this experiment, adult B. tabaci were counted
on melon leaves from early March till 20th May. The
population of B. tabaci increased with the growth of
melon creepers. Two population peaks, during 4th
week of March and 3rd week of April, were
observed. The highest mean density was recorded
during the 2nd peak.
Density of adults B. tabaci was significantly
higher at the 2nd node with 14.90±1.54
individuals/node than 6.36±0.94 at 3rd node and
6.54±0.75 at 5th node. Mean total density of the
adults was 6.56±1.34 at 2nd node, while lower
number 2.52±0.55 individuals at 5th node. The result
indicates that adults prefer 2nd nodes of leaves,
because they are fresh and broader in size.
Naranjo and Flint (1995) reported that B. tabaci
adults on the top stratum of the plant were fairly
uniformly distributed over leaves from main stem of
2-7 nodes, but adults were abundant at 5th node
leaves. Musuna (1986) stated that B. tabaci adult
sampling unit comprises the first two fully expanded
main terminal leaves and one leaf at mid level of the
plant.
Like the indirect and direct count of adult B. tabaci
on the melon leaves, the data in Table-I shows similar
trend of population growth, i.e., two population peaks:
first in 4th week of March and the 2nd in 3rd week of
April. The data of the number of B. tabaci adults was
significantly different at the two timings (T=3.73,
P<0.01). Mean total density of adult fly at 8.00 am
was 6.56±1.34, while 4.03±0.83 at12.00 noon. The
present results are in agreement with those of Gerling
and Horowitz (1984) They reported that direct
counting should be made early in the morning when
adults are least mobile. Musuna (1986) found similar
trend of B. tabaci adult sampling. According to him B.
tabaci adult count was higher during 9.00-12.00h than
during 17.00- 18.00h, although the differences were
not significant at P=0.05.
Table-1. Mean density/leaf of B. tabaci recorded
on 2nd node leaves at 8.00 a.m. and 12.00 noon
on melon creeper during spring season.
Observation
Date
04- March
11
18
25
01- April
08
15
22
29
06- May
13
20
Mean
SE
2nd node
8.00 a.m.
Mean ± SE
2.00 ± 0.29
4.18 ±0.61
8.54 ±0.88
13.45 ±1.86
4.00 ±0.52
9.81 ±0.96
11.81 ±1.56
14.90 ±1.54
3.54 ±0.67
2.72 ± 0.47
2.00 ± 0.35
1.81 ±0.31
6.56
1.34
2nd node
12.00 noon
Mean ± SE
1.27 ±0.24
2.72 ±0.38
4.90 ± 0.74
7.45 ± 0.89
2.90 ±0.58
8.09 ±0.61
4.36 ±0.56
10.1 ±0.82
2.54 ±0.35
1.45 ±0.23
1.45 ±0.23
1.09 ±0.20
4.03
0.83
Lanjar, A.G. et al.
54
REFERENCES
ATTIQUE, M.R., MUHAMMED RAFIQ, ABDUL
GHAFFAR, ZAHOOR AHMAD AND A.I.
MOHYUDDIN (2003). Hosts of Bemisia
tabaci (Genn.) (Homoptera: Aleyrodidae) in
cotton areas of Punjab, Pakistan. Crop
Protection 22 (5): 715-720.
LANJAR, A.G. AND H.A. SAHITO (2005). Effect of
some spring hosts on the life cycle of whitefly,
Bemisia tabaci (Genn.) Pakistan j. entomol.
Karachi 20 (1-2): 13-18.
MELAMED-MADJAR, V., S. COHEN, M. CHEN, S.
TARN AND D. ROSILIO (1982). A method for
monitoring Bemisia tabaaci and timing of spray
application against the pest in cotton fields in
Israel. Phytoparasitica. 10: 85-91.
BERLINGER, M.J. (1980). A yellow sticky trap for
whiteflies: Trialeurodes vaporariorum and
Bemisia tabaci (Aleyrodidae). Ent. Experim.. 27:
98-102.
MOUND, L.A. (1965). Effect of leaf hair on cotton
whitefly populations in the Sudan Gezira. Empire
Cotton Growing Review. 42: 33-40.
BUTLER, G.D. JR., T.J. HENNEBERRY AND W.D.
HUTCHISON (1986). Biology, sampling and
population dynamics of Bemisia tabaci. Agric.
Zool. Rev.l:167-195.
MOUND, L.A. AND S.H. HALSEY (1978). Whitefly of
the world: A systematic catalogue of Aleyrodidae
(Homoptera) with host plant and natural enemy
data. British Museum (Natural History). John
Wiley & Sons, London. 340 pp.
BYRNE, D.N. (1999). Migration and dispersal by the
sweet potato whitefly, Bemisia tabaci. Agricultural
and Forest Meteorology 97 (4): 309-316.
BYRNE, D.N., P.K. VON BRETZEL AND C.V.
HOFFINAN (1986). Impact of trap design and
placement when monitoring for the bandedwinged whitefly, and sweet-potato whitefly
(Homoptera: Aleyrodidae). Environ. Ent. 15:
300-304.
COHEN, S. (1982). Control of whitefly vector of virus
by color mulching. In pathogens, Vectors and
Diseases, approaches to Control ( Harris K.F
and K. Maramoroches, Eds.). Academic Press,
New York. pp. 45-56.
FOWLER, H.D. (1956). Some physiological effects
of attack by whitefly (Bemesia gossypiperda)
and of spraying parathion on cotton in the Sudan
Gezira. Empire Cotton Growing Review. 33:
288-299.
GERIING, D. AND A.R. HOROWITZ (1984). Yellow
traps for evaluating the population levels and
dispersal patterns of Bemisia tabaci (Gennadius)
(Homoptera: Aleyrodidae). Ann. Ent. Soc. Amer.
77: 753-759.
GREATHEAD, A.H. (1986). Host plants. In Bemesia
tabaci. A Literature Survey (M.J.W.Cock, Ed.).
C.A.B. Intern. Inst. Biolo. Cont, U.K. pp.17-25.
GUSMÃO, M.R., M. C. PICANÇO, J.C. ZANUNCIO,
D.J.H. SILVA AND J.A.F. BARRIGOSSI (2005).
Standardised
sampling
plan
for Bemisia
tabaci (Homoptera: Aleyrodidae) in outdoor
tomatoes. Scientia Horticulturae 103 (4):403412.
MUSUNA, A.C.Z. (1986). A method for monitoring
whitefly, Bemisia tabaci (Genn.) in cotton in
Zimbabwe. Agric. Ecosys. Environ. 17: 29-35.
NARANJO, S.E AND H.M. FLINT (1994). Spatial
distribution of pre-imaginable Bemisia tabaci
(Homoptera: Aleyrodidae) in cotton and
development of fixed precision sequential
sampling plants. Environ. Ent. 23 (2): 254-266.
OHNESORGE, B., AND G. RAPP (1986). Monitoring
Bemisia tabaci: A review. Agric. Ecosys. and
Environ. 17 (1/2): 21-27.
SEQUEIRA, R.V AND S.E. NARANJO (2008).
Sampling and
management
of Bemisia
tabaci (Genn.) biotype B in Australian cotton.
Crop Protection 27 (9): 1262-1268
SHARAF, N.S. (1982). Determination of the proper
height, direction, position and distance of a
yellow sticky trap for monitoring adult sweet
potato whitefly populations Bemisia tabaci
Genn., (Homoptera: Aleyrodidae). Dirasat. 9:
169-182.
YOUNGMAN, R.R., N.C. TOSANCO, V.P. JONES,
K. KIDO AND E.T. NATWIK (1986). Correlation
of seasonal trap counts of Bemisia tabaci
(Homoptera: Aleyrodidae) in southeastern
California. J. Econ. Ent. 79: 67-70.
POPULATION DYNAMICS OF GROUND WATER BREEDING OF CULEX
MOSQUITOES OF KARACHI AND THATTA DISTRICT
TANVEER1 FATIMA SIDDIQUI AND NAQVI2, S.N.H.
1
Department of Zoology, Federal Urdu University of Arts, Sciences and
Technology, Gulshan-e-Iqbal Campus, Karachi-Pakistan.
2
Department of Pharmacology, Baqai Medical University, Super Highway,
Toll Plaza, Karachi-Pakistan.
ABSTRACT
A general survey of population of mosquito larvae commonly found in Karachi and Thatta district was
carried out. The mosquito larvae of six (6) different species were collected from different ground water
habitats, during January 2004 to December 2004. Samples were collected, identification, progressive
changes, their abundance and faunistic association noted, are included.
It is to be noted that larval fauna depends not only on the type of the habitat but also on the changes in
the seasons, resulting in the replacement of summer with winter species.
Key words: Population, Dynamics, Breeding, Culex Mosquito, Karachi, Thatta District.
INTRODUCTION
Mosquitoes are very important vectors and
ectoparasite for human and animals. Insect-born
diseases, such as malaria, yellow-fever, lymphatic
filariasis, dengue, encephalitis, are transmitted by
mosquitoes.
Up till now little systematic efforts has been
made to study the culicidae of Pakistan. The fauna
of British India dealing with anopheline (Christophers
1933) and culicine (Barnard 1934) mosquitoes still
remains the standard reference work. An excellent
survey work done on the culicidae of Ceylon, India,
and Pakistan has been given by Qutubuddin (1960).
The culicidae of Pakistan remain poorly known
except for the anopheline mosquitoes, which are
better known because of their involvement in the
transmission of malaria. Information about the
culicine mosquitoes of Pakistan, special reference to
Karachi is still very meager. However studies on
bionomics, filariasis and cytogenetics have added
some information concerning the distributional
record of mosquitoes Nasiruddin 1952; Aslamkhan
and Salman 1969; Baker and Aslamkhan 1969;
Aslamkhan and Wolf 1971).
Adequate
knowledge
of
disease-vector
population dynamics is of paramount importance in
the understanding of the epidemiology of any insectborn disease.
The purpose of present survey of larval breeding
site in the Karachi and Thatta District of Pakistan
was performed to delineate the range of conditions
under which ground water mosquitoes breed, thus
providing some indication of the oviposition
preferences of the females and population of various
prevailing species.
Water samples were collected from different
localities of all five districts of Karachi and Thatta
district. All promising sites were first inspected for
the presence of larval mosquitoes, negative samples
were not analyzed. Larval abundance was estimated
by counting the number of culicine larvae found in 10
to 20 dips with standard dipper. The number of dips
varied with the size of the habitat, the density and
dispersion of the larvae. Less dips were taken in
small homogenous habitats such as ground pools,
while more dips were taken in heterogenous
environments such as large ponds etc., where
mosquito abundance varied considerably from dip to
dip.
All collection were taken back to the laboratory
where a maximum of 10 slides of “different looking”
larvae were made and identified (WHO 1975). The
remainder of the collection was reared to adult,
identified and counted. The percentage composition
of the total specimens identified including both larval
slides and emerging adults was used to estimate the
mean number of larvae per dip for each species of
Culex mosquitoes.
Temperature (ºC), pH, dissolved oxygen of each
sample were also recorded. Temperature were taken
56
Tanveer Fatima Siddiqui
with an ordinary thermometer at the edge of the
habitat where larvae were collected.
pH was recorded by pH meter, Dissolved
oxygen mg/L was estimated by Wrinklers method in
accordance with the procedure of Taras et al.
(1971).
Mosquito larvae were sampled throughout the
year 2004; the immature stages of Culex genus were
the most abundant mosquito larvae in the study
areas. Six (6) Culex mosquito species were collected
in the study area. Both immature and adult of these
species were identified; these species are Culex
bitaeniorhynchus, Culex fusco, Culex p. fatigans,
Culex tritaeniorhynchus, Cx. pseudorishnui and Cx.
vagans.
In the study area different breeding habitats
were characterized. For convenience, the habitats
samples have been classified into six categories
defined below.
1. Pond – Large body of water, usually permanent,
adjacent to some small pockets of population.
2. Pool – Small body of standing water, probably
temporary (i.e., dries during the hot months or in
dry season), filled by rain or run-off water.
3. Borrow pit – Small depression, excavated for
construction materials; difficult to tell old borrow
pit from ground pools.
4. Ditches – Road side drain to carry off rain water
or any other sources.
5. Catch basin – Cement tanks, usually filled with
water, when sufficient water was not available.
6. Artificial container – Tyre puncture tubs,
overhead tanks, buckets, flower pots etc.
Physico-chemical conditions (Temp, pH, and
Dissolved oxygen) for each of the six habitats have
been analyzed. Temperature varied mostly due to
seasonal changes. In general, smaller bodies of
water showed the greatest variation than larger
bodies. Temperature has two peaks with higher
temperature in May-June and September-October.
Slight changes in pH and dissolved oxygen were
also observed due to change in the weather
conditions, or filling of habitats by rain or sewerage
water.
A total of 19414 mosquito larvae were collected
and examined during 2004, collectively from all six
Districts out of 19414 larvae 15512 larvae were from
Culex genus and rest belong to Anopheles and
Aedes genus Table-1 shows that Culex population is
most prevalent among all the three genera.
Detail population is given in (Table-2, and
Figure-1 and 2).
A total of 15512 Culex mosquito larvae were
collected from all Districts, comprising six different
specimens (Table-2, and Figure 1 and 2). In the genus
Culex, the Cx. p. fatigans was the most common, and
showed highest percentage, i.e. 47.43%. The second
highest was of Cx. tritaeniorhynchus i.e. 34.20%.
Culex bitaeniorhynchus and Culex vagan were found
in minimum percentage with 3.27% and 2.68%,
respectively.
Seasonal fluctuation of different Culex species
during twelve months, i.e. from January 2004 to
December 2004 (Graph and Data-1). It shows that
Culex p. fatigans was the most common throughout
the year, but inversely related to temperature, while
Culex tritaeniorhynchus, Culex pseudovishnui and
Culex fuscocephalus directly related to temperature.
According to data and graph population of
Cx. p. fatigans increased during November 2004 and
January 2004. As temperature decreases population
frequently increases.
With the analysis of different species and
breeding habitats, it was found that Cx. bitae
collected from ponds, pools and ditches.
Sirivanakarn (1976) has elevated the dark form of
Cx. bitae. In the present survey, light and dark forms
were not routinely separated. This species mostly
breed in clean water with spirogyrs.
Cx. fuscocephalus (Theobald, 1902) collected
only from ponds and pools with a dense cover of
duckweed (Lemnaceae) or dark-coloured water with
decaying leaves. Cx. p. fatigans (Wiedemann, 1928)
mostly collected from ponds, pools and catch basins,
this mosquito species is inversely correlated with
temperature during winter, population densities
frequently exceeded 400 larvae/dip in polluted pool
habitats. Cx. pseudorishnui (Colles, 1957) breed
almost all different type of habitats except catch
basin and polluted water. This species is positively
correlated with temperature. This species frequently
associated with Cx. tritaeniorhynchus (e.g., Colless,
1957; Reuben 1971). This species mostly occurs in
clean-water habitats. Cx. tritaeniorhynchus was most
frequently collected species during monsoon and
post monsoon seasons. Positively related to
temperature and inversely related to dissolved
oxygen. During the summer Cx. tritaeniorhynchus
breeding in all ground water habitats, especially with
grasses and decaying vegetation were high.
Cx. vegans mostly found in permanent ponds. This
species mostly found in Lahore (Barnard 1934), but
in few cases in Karachi.
In all District of Karachi and Thatta District
definite
mosquito-habitat
associations
were
determined which were modified by increases in
temperature, and organic pollution. Seasonal
temperature changes resulted in the shift of fauna
e.g., Cx. p. fatigans increases in abundance during
winter mean cold weather and Cx. tritaeniorhynchus
increased during summer-season.
Population dynamics of ground water breeding of Culex mosquitoes of Karachi & Thatta District
57
58
Population dynamics of ground water breeding of Culex mosquitoes of Karachi & Thatta District
Graph and Data-1
59
60
REFERENCES
ASLAMKHAN, M., AND BAKER, R.H., (1969).
Karyotypes of some Anopheles, Filalbia, and
Culex mosquitoes of Asia. Pakistan J. Zool., 1:
107.
ASLAMKHAN, M., AND SALMAN, C., (1969). The
bionomics of the mosquitoes of the Changa
Manga National Forest, West Pakistan. Pakistan
J. Zool. 1: 183-205.
ASLAMKHAN, M. & WOLF, M.S. (1971). Rural
bankroftian filariasis in two villages in Dinajpur
District, East Pakistan, II. Entomological
investigation. Am. J. Trop. Med. Hyg., 20.
REISEN, W.K. AND SIDDIQUI, T.F., (1977). The
influence of consepecific immature on the
oviposition preferences of Anopheles stephensi
Liston and Culex tritaeniorhynchus Giles
(Diptera: Culicidae). J. Med. Ent.
REUBEN, R. (1971). Studies on the mosquitoes of
North Arcot District, Madras State, India. Part 5.
Breeding places of the Culex vishnui group of
species. J. Med. Ent., 8: 363-66.
SIRIVANAKARN, S., (1976). Medical entomological
studies III. A revision of the subgenus Culex in
the oriental region (Diptera: Culicidae). Contr.
Amer. Ent. Int., 12: 1-72.
BAKER, R.H., AND ASLAMKHAN, M., (1969).
Karyotype of some Asian mosquitoes of the
subfamily Culicinae (Diptera; Culicidae). J. Med.
Ent., 6: 44-52.
TARAS, M.J., GREENBERG, A.E., HOAK, R.D.
AND RAND, M.C., (1971). Standard methods for
the examination of water and waste water. 13th
Edition, Amer. Publ. Hlth. Assoc. Washington,
D.C. p. 874 p.
BARNARD, P.J., (1934). The fauna of British India,
including Ceylon and Burma. Diptera Vol. 5,
Family Culicidae. Tribes Megarhinini and
Culicini. London, XXX VIII + 463 pp. London.
THEOBALD, F.V. (1902). A short description of the
Culicidae of India, with description of new
species of Anopheles. Proc. Roy. Soc., 69, 367394.
CHRISTOPHERS, S.R., (1933). The fauna of British
India, including Ceylon and Burma. Anophelini,
London, XII + 371 pp. London.
WADA, Y., M. MOGI, AND J. NISHIGAKI, (1971).
Studies on the population estimation for insects
of medical importance. IV, A method for the
estimation of relative density of Culex
tritaeniorhynchus larvae in the whole paddyfields of an area. Trop. Med. 13 (2): 86-93.
COLLESS, D.H., (1957). Note on the Culicine
mosquitoes of Singapore. III. Larval breeding
places. Ann. Trop. Med. Parasit. 51: 102-16.
NASIRUDDIN, M. (1952). Mosquitoes breeding in
tree-holes and bamboo stumps in Dacca (East
Pakistan, Bangladesh). Pakistan J. Hlth. 2: 110112.
QUTUBUDDIN, M., (1960). The mosquito fauna of
Kohat-Hangu Valley, West Pakistan. Mosquito
News, 20: 355-361.
WHO (1975). Manual of practical entomology in
malaria. Part II. Methods and techniques. Wld.
Hlth. Org., Geneva, Switzerland, 191 pgs.
LEVELS OF DENGUE FEVER VIRUS CONTROL: THE FFECTIVENESS
AND VASTNESS OF CONTROLLING POWER BOUNDARIES
OF THESE LEVELS
RAJPUT MUHAMMED TARIQ1 & S. SALAHUDDIN QADRI2
1
MAH Qadri Biological Research Centre, Room No.11,
University of Karachi, Karachi-75270, Sindh-Pakistan
2
Department of Zoology, Jamia Millia Degree College, Malir Karachi-Pakistan
ABSTRACT
Each (any) problem has some levels (steps) to solve that problem. Usually a problem is solved step by
step, but some times, in special and the most important case the particular problem is solved by taking
all steps simultaneously and not by step by step. This type of situation happens there, where the
boundaries of vastness of that problem grows by the passage of time. It means when all steps are
necessary to be taken and there is a shortage of time to solve that problem, especially when the
problem is growing with the passage of time. The Dengue Fever Virus (DFV) problem is being spread
by a mosquito genus, the Aedes. The problem is increasing day-by-day not only in Pakistan but also in
the world by the passage of time. Therefore the levels (steps) with their controlling power and
effectiveness boundaries are discussed in detail.
Key words: Aedes, DFV, Levels, Control, Effectiveness, Boundaries.
INTRODUCTION & DISTRIBUTION
The Aedes aegypti & Ae. albopictus have the
global distribution and both are the confirmed vector
(causative agents) of DFV. Barraud (1934) has
reported Aedes aegypti in Pakistan since last 75
years. Naqvi (1992) presented a survey report of
mosquitoes and houseflies of Karachi regions from
12 stations selected in the region of Karachi.
Suleman et al. (1993) reported 8 species of Aedes in
Pakistan from Peshawar, Charsadda, Mardan &
Swabi viz. Ae. albopictus, capius, lineatopenis,
pseudotaeniatus, pulchriventer, vittatus, uniliniatus &
walbus. The ninth species of Ae. aegypti has been
present long before in different areas of Pakistan as
reported by Barraud (1934), especially in Punjab &
Sindh. Kamimura et al. 1986, WHO (1989) also
reported by Chan et al. (1995). Suleman (1996),
Naqvi, et al. (1997), Tariq & Zafar (2000), Tariq &
Qadri (2001), Qadri et al. (2007).
MATERIALS & METHODS
Literature survey, observations for controlling the
vector of DFV, Medical strategies in Hospitals of
Karachi in 2006, 2007 & 2008.
RESULTS & DISCUSSIONS
There are three basic levels to control the
Dengue & Malaria vector and other mosquitoes as
well.
(1) Ground level: (Larval stage/Breeding stage).
This stage is restricted to some/few places
where they (Larvae) breed they may be control and
destroyed easily, as they are non-flyer, remain at
particular place in limited area. They may be
controlled by destroying water in which they are
contained, by putting Bt., fishes & Lemna plants in
large area water which can not be destroyed or dried
up containing these larvae. These larvae usually
breed in small pots, plastic containers, cans, drums,
earthen pots, over-head & ground water tanks,
present in the human populated area. As they are
present in hundreds in small plastic containers & in
thousands number in the stored water for checking
the puncture tubes at the puncture tyre shops as
reported by Tariq & Zafar (2000). According to their
report 11.2% puncture tyre shops were recorded
positive for Aedes mosquito eggs larvae, pupae &
adults. If awareness is created among the public
through media then the destruction of these breeding
points & places is sure. These breeding points are
easy to destroy and cheep or less expensive as
compared to spray and other methods. Only the
manpower is needed to do work against breeding
points themselves, after awareness by media and
other sources.
(2) Air level: Adult stage/flying stage of
mosquito.
In the adult stage the mosquitoes are flyer and
spread far and wide in the area, they are breeding.
As they fly from one place to another place their
boundaries of presence increase, but the power of
control through spray method decreases, due to
which the effectiveness of the control strategy also
decrease. At the same time expenses to control
them increases ultimately due to the use of
insecticide, spray machine, sprayer, vehicle, driver,
spray-man, diesel, etc. for spraying. The spray is not
Tariq et al.
62
done simultaneously in the area (Say Karachi city) it
is done area by area, stepwise due to this reason the
mosquitoes shift from the sprayed area to the other
areas, due to which they are not killed but remain
alive. The other reason is that the only selected area
is sprayed not the whole city or area.
Thus the spreading boundaries (as they are
flyer) of mosquitoes in this stage is increased, with
the increased expenses but the boundaries of
controlling power to these flying mosquitoes is
decreased resulting in less effectiveness of control,
as compared to ground level control of mosquitoes
and few mosquitoes may be killed in this level by
spray. All the people or most of the people may be
infected in this stage.
(3) Clinical/Hospital level: Infection stage by
mosquitoes.
This is the stage in which only infected persons
are brought to Clinic/Hospital, after the diagnostic
report and tests, the patients are given platelets in
points. The expenses in this stage increases more
and more as compared to second stage due to the
high cost of platelet bags, platelets, Clinic/Hospital
charges, diagnostic test charges, etc. In this level
only few hundred people are given treatment, only
those who can reach Clinic or Hospital.
The transmission of virus by the mosquitoes is
continue and is increasing day-by-day by the
passage of time during infection season (out break)
Jawad et al. (2001), Kautner et al. (1997). The
number of patients increases an daily basis,
because only the patients are given concentration
but not a single mosquito is killed in this level. The
mosquito which is infecting the person to be patient
is left alive, free and uncontrolled, to save the life of
the patient is left the only option for us. So here the
infective boundaries by the mosquitoes are
unlimited, the expenses are very high as compared
to second level and the control of the causative
agents is zero.
CONCLUSION
It may be concluded that all 3-steps are
necessary to control the situation of this problem
currently. Unfortunately no concentration is given to
the 1st level, only formalities are being completed for
the second level and full/all concentration is being
given to the 3rd level in which causative agent control
is zero. If this strategy will not be changed then how
we can solve this problem faced by us during last
years and still are facing.
REFERENCES
BARRAUD, P.J., (1934). The fauna of British India,
including Ceylon and Burma. Diptera Family
Culicidae. Tribes Megarhimini and Culicinae,
Vol. 5, Taylor and Francis, London.
CHAN, Y.C., SALAHUDDIN, N.I. AND KHAN, J.
(1995). Dengue Haemorrhagic Fever out break
in Karachi, Pakistan. Trans. R. Soc. Trop. Med.
Hyg. 88: 619-620.
JAWAD, K.A., MASOOD, S., TASSAWAR, H.,
INAM, B., WAHEED UZ ZAMAN, T. (2001). Out
break of Dengue Haemorrhagic fever in Karachi.
Pakistan Armed Forces Medical Journal 51(2):
94-98.
KAMIMURA, K., TAKASU, T., AHMED, A. AND
AHMED, A. (1986). A survey of mosquitoes in
Karachi area. Pakistan J. Pak. Med. Assoc., 36:
182-187.
KAUTNER, INGRID, ROBINSON, MAX MD.
KUHNLE, URSULA MD, (1997). “Dengue virus
infection: Epidemiology, pathogenesis, clinical
presentation, diagnosis, and prevention”. The
Journal of Pediatrics. Vol. 131(4): 516-524.
NAQVI, S.N.H. (1992). Survey and determination of
resistance in mosquitoes and houseflies of
Karachi region. PSF-Project No. S-KU/Bio-161,
pp. 1-132.
NAQVI, S.N.H., TABASSUM, R., AHMAD, I. AND
FATEMA, T. (1997). Seasonal population
fluctuation of mosquito larvae in different
locations of Karachi region. Bull. Pure and
Applied Sci. 16A (1-2): 15-19.
QADRI, S.S., TARIQ, R.M. AND AHMED, I. (2007).
Dengue Kee Wapsee. Global Science, Nov.
2007, 21-26.
SULEMAN, M. AND SHAFQAT, K. (1993). Notes on
Aedine Mosquitoes as Diurnal Pests of Human
skin Abbottabad Area. Pakistan Journal of
Zoology 25(3): 253-260.
SULEMAN, M., ARSHAD M., AND KHAN, K. (1996).
Yellow Fever Mosquito (Diptera: Culicidae)
introduced into Landi Kotal, Pakistan tire
importation. J. Med. Ent. 33: 689-693.
SULEMAN, M., KHALID, M. AND KHAN, S., (1993).
Ecology of mosquitoes in Peshawar adjoining
areas species composition and relative
abundance. Pakistan J. Zool., 25: 321-328.
TARIQ, R.M. & ZAFAR, S.M.N. (2000). Why the
population of Dengue vector mosquitoes is
increasing day-by-day in Karachi and other
areas of Sindh, Pakistan? Pakistan j. entomol.
Karachi. 15(1&2): 7-10.
WHO, (1989). Geographical distribution of
arthropods borne diseases and their principle
vectors, WHO/Vector Biology and control
Division, Geneva, No. WHO/VBC/89-967.

front page 08.BMP - Pakistan Journal of Entomology Karachi

Transcription

Documents pareils

programme

EMBASSY OF PAKISTAN IN FRANCE PHOTO

09 : V : 2013

changing family issues - Hu

Russian Academician Alexander Granberg

Is India a Major Power?

Univ.-Prof. Dr. Husain Kassim