Journal of the American Mosquito Control Association, 1996
The effects of laundering and time of exposure on the insecticidal activity of military uniform f... more The effects of laundering and time of exposure on the insecticidal activity of military uniform fabric impregnated with permethrin at 0.125 mg active ingredient (AI)/cm2 are reported from susceptibility tests with laboratory-reared mosquitoes, Culex pipiens, and sand flies, Phlebotomus papatasi. Knockdown/mortality resulting from exposure of groups of female insects for periods of 1, 3, 5, 7, and 10 min was recorded and compared among 5 different treatment/wash groups (untreated/unwashed, treated/unwashed, treated/1-wash, treated/2-wash, treated/3-wash). Laundering was by machine washing with detergent and warm water followed by hot-air machine drying. Post-exposure assessments at intervals from 0 to 60 min and at 24 h showed that knockdown for each exposure time and wash group was initially low but increased steadily during the first hour post-exposure. Sand flies were less sensitive than mosquitoes to knockdown during the first 60 min after contact with treated/unwashed fabric; ho...
An intraspecific study on Phlebotomus papatasi, the main proven vector of Leishmania major among ... more An intraspecific study on Phlebotomus papatasi, the main proven vector of Leishmania major among the members of the subgenus Phlebotomus, was performed. The internal transcribed spacer 2 (ITS 2) of rDNA and the ND4 gene of mt DNA were sequenced from 26 populations from 18 countries (Albania, Algeria, Cyprus, Egypt, Greece, India, Iran, Israel, Italy, Lebanon, Morocco, Saudi Arabia, Spain, Syria, Tunisia, Turkey, Yugoslavia and Yemen), and compared. Samples also included three other species belonging to the subgenus Phlebotomus: P. duboscqi, a proven vector of L. major in the south of Sahara (three populations from Burkina Faso, Kenya and Senegal), P. bergeroti, a suspected vector of L. major (three populations from Oman Sultanate, Iran and Egypt), and one population of P. salehi from Iran. A phylogenetic study was carried out on the subgenus Phlebotomus. Our results confirm the validity of the morphologically characterized taxa. The position of P. salehi is doubtful. Variability in P. papatasi contrasts with that observed within other species having a wide distribution like P. (Paraphlebotomus) sergenti in the Old World or Lutzomyia (Lutzomyia) longipalpis in the New World. Consequently, it could be hypothesized that all populations of P. papatasi over its distribution area have similar vectorial capacities. The limits of the distribution area of L. major are correlated with the distribution of common rodents acting as hosts of the parasites.
Ivermectin (IVM) is a chemically modified macrocyclic lactone of Streptomyces avermitilis that ac... more Ivermectin (IVM) is a chemically modified macrocyclic lactone of Streptomyces avermitilis that acts as a potent neurotoxin against many nematodes and arthropods. Little is known of IVM's effect against either blood-feeding Phlebotomus sand flies, or the infective ...
Phlebotomus papatasi sand flies are major vectors of Leishmania major and phlebovirus infection i... more Phlebotomus papatasi sand flies are major vectors of Leishmania major and phlebovirus infection in North Africa and across the Middle East to the Indian subcontinent. Population genetics is a valuable tool in understanding the level of genetic variability present in vector populations, vector competence, and the development of novel control strategies. This study investigated the genetic differentiation between P. papatasi populations in Egypt and Jordan that inhabit distinct ecotopes and compared this structure to P. papatasi populations from a broader geographical range. A 461 base pair (bp) fragment from the mtDNA cytochrome b (cyt b) gene was PCR amplified and sequenced from 116 individual female sand flies from Aswan and North Sinai, Egypt, as well as Swaimeh and Malka, Jordan. Haplotypes were identified and used to generate a median-joining network, F values and isolation-by-distance were also evaluated. Additional sand fly individuals from Afghanistan, Iran, Israel, Jordan, L...
Journal of the American Mosquito Control Association, Jun 1, 2004
The effectiveness of 1-octen-3-ol (octenol) as an attractant for collecting medically important p... more The effectiveness of 1-octen-3-ol (octenol) as an attractant for collecting medically important psychodids has never been reported. This study evaluated the effects of carbon dioxide (CO2) and octenol released at 2 rates, individually and in combination, as attractants for adult sand flies in a small village in southern Egypt. Four sand fly species were collected: Phlebotomus papatasi, P. sergenti, Sergentomyia palestinensis, and S. schwetzi. Only P. papatasi was collected in numbers sufficient to allow statistical analysis. This study reaffirms that CO2 is an effective attractant for female P. papatasi and also demonstrates that neither male nor female P. papatasi respond to octenol alone. Additionally, no synergistic attractancy for either females or males was observed when CO2 and octenol were combined.
Journal of the American Mosquito Control Association, 2001
The purpose of this study was to determine the baseline susceptibility of 4 species of phlebotomi... more The purpose of this study was to determine the baseline susceptibility of 4 species of phlebotomine sand flies from North Africa and the Middle East to various insecticides. Susceptibility was determined using the World Health Organization test kits for measuring resistance in mosquitoes exposed to insecticide-impregnated papers. Fifty, 90, and 99% lethal doses were calculated for bendiocarb, cyfluthrin, DDT, malathion, permethrin, and resmethrin on Phlebotomus bergeroti, P. langeroni, P. papatasi. and P. sergenti. The least toxic insecticide to all species was DDT, followed by malathion and permethrin in order of increasing toxicity. Cyfluthrin was the most toxic to P. langeroni and P. papatasi, followed by resmethrin and bendiocarb in order of decreasing toxicity. Resmethrin exhibited the highest toxicity to P. bergeroti followed by cyfluthrin and bendiocarb, whereas bendiocarb was most toxic to P. sergenti, followed by cyfluthrin and resmethrin in order of decreasing toxicity. An attempt was made to obtain data for deltamethrin, but close response data were insufficient to determine regression lines for this insecticide on these species. However, analysis of preliminary data indicated that deltamethrin is highly toxic to these sand flies.
Country-wide surveys forsand fly vectorsotLeishmania conductedduring 1988 and 1989 intheRepublic ... more Country-wide surveys forsand fly vectorsotLeishmania conductedduring 1988 and 1989 intheRepublic ofDjibouti. Standardized, consistentsamplingby oiledpapertraps concentrated within ten towns and encampments in four distinct ecological/altitudinal (coastal plain, 0-100 m; inland plateau, 400-750 m; mountain valley, 1,000 m; highland forest, >1,500m) and timedduring three seasonal periods (cool-dry, October-November; cool-moist, January-February; hot-dry, June-July). A totalof8,492 specimens,representing 19speciesandsubspecies identified. Phlebotomusspecieswith proven potentialability to transmitLeishmaniaaccountedfor22 percent (1,902/8,492) ofthe identified collection and comprised 15 to 39 percent of each seasonal sample. Phlebotomus alexandri and Phlebotomus bergeroti amongthemost andwidely distributed species,andmadeup62and 36percent,respectively,ofthePhlebotomuscollection. PhlebotomussergentiandPhlebotomusorientally also found,but atlownumbers and fromfewer locations. Seasonal ...
Sand fly saliva can modulate the host hemostatic, inflammatory and immunomodulatory responses and... more Sand fly saliva can modulate the host hemostatic, inflammatory and immunomodulatory responses and pre-exposure to saliva or bites of uninfected sand flies protect against leishmaniases in animal models. A number of salivary proteins involved in this protection were identified. However, polymorphisms in sequence and expression of sand fly salivary gland genes can modulate disease outcome. In this study, expression polymorphisms of nine salivary gland genes were assessed in three natural populations of Phlebotomus papatasi, the main vector of Leishmania major. Sand flies were collected at different locations in Egypt (Aswan and North Sinai) and Jordan (Sweimah), during different times of the year (i.e., early and late during the sand fly season) for two years. Expression of the salivary genes SP12, SP14, SP28, SP29, SP30, SP32, SP36, SP42, and SP44 were assessed by Real-Time PCR, and potential seasonal effects were evaluated. Our results indicate that an increase in mRNA abundance tow...
Journal of the American Mosquito Control Association, 2001
We report the 1st collection of Phlebotomus sergenti, a vector of the cutaneous and visceralizing... more We report the 1st collection of Phlebotomus sergenti, a vector of the cutaneous and visceralizing forms of Leishmania tropica, from southern Egypt. Four female and 1 male P. sergenti were collected from unlit Centers for Disease Control light traps placed in a village on the Nile River, 6 km north of Aswan, Egypt, during studies conducted from 1998 to 1999. This extends the known distribution of this species farther south in Egypt than previously recorded.
Journal of the Egyptian Society of Parasitology, 2007
Phlebotumus kazeruni, a blood-feeding, xerophilic sand fly species found broadly throughout North... more Phlebotumus kazeruni, a blood-feeding, xerophilic sand fly species found broadly throughout North Africa and Western Asia, is a suspected vector of cutaneous leishmaniasis (CL). Following successful laboratory colonization of this species, we employed the murine (BALB/c) infection model to determine whether our Sinai strain of P. kazeruni was able to successfully acquire, develop, and transmit a Sinai strain of Leishmania major. Groups of female sand flies were fed 1) by membrane, hamster blood containing culture-produced L. major promastigotes, 2) by membrane, hamster blood containing a suspension of L. major tissue amastigotes, and 3) directly upon L. major lesions in BALB/c mice. Samples of blood-fed sand flies from each group were dissected on selected days post-feeding and examined by light microscope for acquired and developing Leishmania infections. Female P. kazeruni acquired viable parasites by the three feeding methods. Development of ingested parasites to infective-stage ...
The fat-tailed gerbil Pachyuromys duprasi is a common burrowing rodent found across the northern ... more The fat-tailed gerbil Pachyuromys duprasi is a common burrowing rodent found across the northern Sahara Desert from Morocco to Egypt. There is overlap in the geographical distribution and ecological habitats of P. duprasi, several Old World Leishmania species, and numerous sand fly vectors of Leishmania, but there are no records that document the natural occurrence of this gerbil with any species of Leishmania or phlebotomine sand fly. Experiments were conducted to determine its potential as a natural host and laboratory animal model for Leishmania major. Captive-born P. duprasi were inoculated subcutaneously (s.c.) in the tail with promastigotes or amastigotes of an Egyptian strain of L. major and monitored for signs of infection. Local swelling and erythema was visible 10-12 days after amastigote inoculation, and within 3-4 weeks swelling had increased tail widths by up to 78%. Infections progressed more slowly and less conspicuously following inoculation with promastigotes. Tissue density of amastigotes in the gerbil's tail lesions after inoculating with either stage of L. major was significantly lower than that produced in the footpads of BALB/c mice by the same parasite and incubation period. Laboratory transmission of L. major to P. duprasi by sand fly bite was demonstrated and acquisition of L. major, by bite, from tail lesions of infected P. duprasi to laboratory-reared Phlebotomus papatasi was also achieved with 10% of biting flies developing promastigote infections. The acquisition and development of L. major infections in P. papatasi after biting an infected P. duprasi and the susceptibility of P. duprasi to L. major delivered at low densities by sand fly bites indicate that fat-tailed gerbils could serve as a natural host and reservoir of L. major.
Journal of the American Mosquito Control Association, 2006
Insecticide and resistance bioassays and microplate assays were performed on Culex pipiens mosqui... more Insecticide and resistance bioassays and microplate assays were performed on Culex pipiens mosquitoes to determine the level and mechanisms of resistance. Culex pipiens larvae were collected from three filariasis-endemic areas of Egypt and reared to adults for subsequent production and testing of F1 generation larvae and adults. Bioassays were performed using World Health Organization (WHO) methods with the diagnostic doses of 6 organophosphate insecticides for larvae and 1 organochlorine (OC), 4 pyrethroid, 2 organophosphate, and 2 carbamate insecticides for adults. Microplate assays were performed to measure levels of beta esterase, acetylcholinesterase, insensitive acetylcholinesterase, oxidases, and glutathione-S-transferase enzymes. Larval bioassay results showed clear indications of resistance to organophosphate insecticides. Adult bioassays also showed widespread, significant resistance to many insecticides from all four classes, including the OC, DDT. The Qalubiya larval population was susceptible only to malathion, whereas Sharkiya larvae were susceptible to malathion, temephos, and chlorpyrifos. On the other hand, larval specimens from Assiut were resistant to all insecticides tested. Larval bioassay results were supported by those of microplate assays in showing elevated levels of glutathione S-transferase in populations from all three areas. In general, microplate results confirmed patterns of resistance observed using bioassays, and mechanisms of resistance were evident for all three areas sampled. Mechanisms of resistance are discussed in relation to microplate and bioassay results for the areas sampled and pesticides used.
Journal of the American Mosquito Control Association, 1996
The effects of laundering and time of exposure on the insecticidal activity of military uniform f... more The effects of laundering and time of exposure on the insecticidal activity of military uniform fabric impregnated with permethrin at 0.125 mg active ingredient (AI)/cm2 are reported from susceptibility tests with laboratory-reared mosquitoes, Culex pipiens, and sand flies, Phlebotomus papatasi. Knockdown/mortality resulting from exposure of groups of female insects for periods of 1, 3, 5, 7, and 10 min was recorded and compared among 5 different treatment/wash groups (untreated/unwashed, treated/unwashed, treated/1-wash, treated/2-wash, treated/3-wash). Laundering was by machine washing with detergent and warm water followed by hot-air machine drying. Post-exposure assessments at intervals from 0 to 60 min and at 24 h showed that knockdown for each exposure time and wash group was initially low but increased steadily during the first hour post-exposure. Sand flies were less sensitive than mosquitoes to knockdown during the first 60 min after contact with treated/unwashed fabric; ho...
An intraspecific study on Phlebotomus papatasi, the main proven vector of Leishmania major among ... more An intraspecific study on Phlebotomus papatasi, the main proven vector of Leishmania major among the members of the subgenus Phlebotomus, was performed. The internal transcribed spacer 2 (ITS 2) of rDNA and the ND4 gene of mt DNA were sequenced from 26 populations from 18 countries (Albania, Algeria, Cyprus, Egypt, Greece, India, Iran, Israel, Italy, Lebanon, Morocco, Saudi Arabia, Spain, Syria, Tunisia, Turkey, Yugoslavia and Yemen), and compared. Samples also included three other species belonging to the subgenus Phlebotomus: P. duboscqi, a proven vector of L. major in the south of Sahara (three populations from Burkina Faso, Kenya and Senegal), P. bergeroti, a suspected vector of L. major (three populations from Oman Sultanate, Iran and Egypt), and one population of P. salehi from Iran. A phylogenetic study was carried out on the subgenus Phlebotomus. Our results confirm the validity of the morphologically characterized taxa. The position of P. salehi is doubtful. Variability in P. papatasi contrasts with that observed within other species having a wide distribution like P. (Paraphlebotomus) sergenti in the Old World or Lutzomyia (Lutzomyia) longipalpis in the New World. Consequently, it could be hypothesized that all populations of P. papatasi over its distribution area have similar vectorial capacities. The limits of the distribution area of L. major are correlated with the distribution of common rodents acting as hosts of the parasites.
Ivermectin (IVM) is a chemically modified macrocyclic lactone of Streptomyces avermitilis that ac... more Ivermectin (IVM) is a chemically modified macrocyclic lactone of Streptomyces avermitilis that acts as a potent neurotoxin against many nematodes and arthropods. Little is known of IVM's effect against either blood-feeding Phlebotomus sand flies, or the infective ...
Phlebotomus papatasi sand flies are major vectors of Leishmania major and phlebovirus infection i... more Phlebotomus papatasi sand flies are major vectors of Leishmania major and phlebovirus infection in North Africa and across the Middle East to the Indian subcontinent. Population genetics is a valuable tool in understanding the level of genetic variability present in vector populations, vector competence, and the development of novel control strategies. This study investigated the genetic differentiation between P. papatasi populations in Egypt and Jordan that inhabit distinct ecotopes and compared this structure to P. papatasi populations from a broader geographical range. A 461 base pair (bp) fragment from the mtDNA cytochrome b (cyt b) gene was PCR amplified and sequenced from 116 individual female sand flies from Aswan and North Sinai, Egypt, as well as Swaimeh and Malka, Jordan. Haplotypes were identified and used to generate a median-joining network, F values and isolation-by-distance were also evaluated. Additional sand fly individuals from Afghanistan, Iran, Israel, Jordan, L...
Journal of the American Mosquito Control Association, Jun 1, 2004
The effectiveness of 1-octen-3-ol (octenol) as an attractant for collecting medically important p... more The effectiveness of 1-octen-3-ol (octenol) as an attractant for collecting medically important psychodids has never been reported. This study evaluated the effects of carbon dioxide (CO2) and octenol released at 2 rates, individually and in combination, as attractants for adult sand flies in a small village in southern Egypt. Four sand fly species were collected: Phlebotomus papatasi, P. sergenti, Sergentomyia palestinensis, and S. schwetzi. Only P. papatasi was collected in numbers sufficient to allow statistical analysis. This study reaffirms that CO2 is an effective attractant for female P. papatasi and also demonstrates that neither male nor female P. papatasi respond to octenol alone. Additionally, no synergistic attractancy for either females or males was observed when CO2 and octenol were combined.
Journal of the American Mosquito Control Association, 2001
The purpose of this study was to determine the baseline susceptibility of 4 species of phlebotomi... more The purpose of this study was to determine the baseline susceptibility of 4 species of phlebotomine sand flies from North Africa and the Middle East to various insecticides. Susceptibility was determined using the World Health Organization test kits for measuring resistance in mosquitoes exposed to insecticide-impregnated papers. Fifty, 90, and 99% lethal doses were calculated for bendiocarb, cyfluthrin, DDT, malathion, permethrin, and resmethrin on Phlebotomus bergeroti, P. langeroni, P. papatasi. and P. sergenti. The least toxic insecticide to all species was DDT, followed by malathion and permethrin in order of increasing toxicity. Cyfluthrin was the most toxic to P. langeroni and P. papatasi, followed by resmethrin and bendiocarb in order of decreasing toxicity. Resmethrin exhibited the highest toxicity to P. bergeroti followed by cyfluthrin and bendiocarb, whereas bendiocarb was most toxic to P. sergenti, followed by cyfluthrin and resmethrin in order of decreasing toxicity. An attempt was made to obtain data for deltamethrin, but close response data were insufficient to determine regression lines for this insecticide on these species. However, analysis of preliminary data indicated that deltamethrin is highly toxic to these sand flies.
Country-wide surveys forsand fly vectorsotLeishmania conductedduring 1988 and 1989 intheRepublic ... more Country-wide surveys forsand fly vectorsotLeishmania conductedduring 1988 and 1989 intheRepublic ofDjibouti. Standardized, consistentsamplingby oiledpapertraps concentrated within ten towns and encampments in four distinct ecological/altitudinal (coastal plain, 0-100 m; inland plateau, 400-750 m; mountain valley, 1,000 m; highland forest, >1,500m) and timedduring three seasonal periods (cool-dry, October-November; cool-moist, January-February; hot-dry, June-July). A totalof8,492 specimens,representing 19speciesandsubspecies identified. Phlebotomusspecieswith proven potentialability to transmitLeishmaniaaccountedfor22 percent (1,902/8,492) ofthe identified collection and comprised 15 to 39 percent of each seasonal sample. Phlebotomus alexandri and Phlebotomus bergeroti amongthemost andwidely distributed species,andmadeup62and 36percent,respectively,ofthePhlebotomuscollection. PhlebotomussergentiandPhlebotomusorientally also found,but atlownumbers and fromfewer locations. Seasonal ...
Sand fly saliva can modulate the host hemostatic, inflammatory and immunomodulatory responses and... more Sand fly saliva can modulate the host hemostatic, inflammatory and immunomodulatory responses and pre-exposure to saliva or bites of uninfected sand flies protect against leishmaniases in animal models. A number of salivary proteins involved in this protection were identified. However, polymorphisms in sequence and expression of sand fly salivary gland genes can modulate disease outcome. In this study, expression polymorphisms of nine salivary gland genes were assessed in three natural populations of Phlebotomus papatasi, the main vector of Leishmania major. Sand flies were collected at different locations in Egypt (Aswan and North Sinai) and Jordan (Sweimah), during different times of the year (i.e., early and late during the sand fly season) for two years. Expression of the salivary genes SP12, SP14, SP28, SP29, SP30, SP32, SP36, SP42, and SP44 were assessed by Real-Time PCR, and potential seasonal effects were evaluated. Our results indicate that an increase in mRNA abundance tow...
Journal of the American Mosquito Control Association, 2001
We report the 1st collection of Phlebotomus sergenti, a vector of the cutaneous and visceralizing... more We report the 1st collection of Phlebotomus sergenti, a vector of the cutaneous and visceralizing forms of Leishmania tropica, from southern Egypt. Four female and 1 male P. sergenti were collected from unlit Centers for Disease Control light traps placed in a village on the Nile River, 6 km north of Aswan, Egypt, during studies conducted from 1998 to 1999. This extends the known distribution of this species farther south in Egypt than previously recorded.
Journal of the Egyptian Society of Parasitology, 2007
Phlebotumus kazeruni, a blood-feeding, xerophilic sand fly species found broadly throughout North... more Phlebotumus kazeruni, a blood-feeding, xerophilic sand fly species found broadly throughout North Africa and Western Asia, is a suspected vector of cutaneous leishmaniasis (CL). Following successful laboratory colonization of this species, we employed the murine (BALB/c) infection model to determine whether our Sinai strain of P. kazeruni was able to successfully acquire, develop, and transmit a Sinai strain of Leishmania major. Groups of female sand flies were fed 1) by membrane, hamster blood containing culture-produced L. major promastigotes, 2) by membrane, hamster blood containing a suspension of L. major tissue amastigotes, and 3) directly upon L. major lesions in BALB/c mice. Samples of blood-fed sand flies from each group were dissected on selected days post-feeding and examined by light microscope for acquired and developing Leishmania infections. Female P. kazeruni acquired viable parasites by the three feeding methods. Development of ingested parasites to infective-stage ...
The fat-tailed gerbil Pachyuromys duprasi is a common burrowing rodent found across the northern ... more The fat-tailed gerbil Pachyuromys duprasi is a common burrowing rodent found across the northern Sahara Desert from Morocco to Egypt. There is overlap in the geographical distribution and ecological habitats of P. duprasi, several Old World Leishmania species, and numerous sand fly vectors of Leishmania, but there are no records that document the natural occurrence of this gerbil with any species of Leishmania or phlebotomine sand fly. Experiments were conducted to determine its potential as a natural host and laboratory animal model for Leishmania major. Captive-born P. duprasi were inoculated subcutaneously (s.c.) in the tail with promastigotes or amastigotes of an Egyptian strain of L. major and monitored for signs of infection. Local swelling and erythema was visible 10-12 days after amastigote inoculation, and within 3-4 weeks swelling had increased tail widths by up to 78%. Infections progressed more slowly and less conspicuously following inoculation with promastigotes. Tissue density of amastigotes in the gerbil's tail lesions after inoculating with either stage of L. major was significantly lower than that produced in the footpads of BALB/c mice by the same parasite and incubation period. Laboratory transmission of L. major to P. duprasi by sand fly bite was demonstrated and acquisition of L. major, by bite, from tail lesions of infected P. duprasi to laboratory-reared Phlebotomus papatasi was also achieved with 10% of biting flies developing promastigote infections. The acquisition and development of L. major infections in P. papatasi after biting an infected P. duprasi and the susceptibility of P. duprasi to L. major delivered at low densities by sand fly bites indicate that fat-tailed gerbils could serve as a natural host and reservoir of L. major.
Journal of the American Mosquito Control Association, 2006
Insecticide and resistance bioassays and microplate assays were performed on Culex pipiens mosqui... more Insecticide and resistance bioassays and microplate assays were performed on Culex pipiens mosquitoes to determine the level and mechanisms of resistance. Culex pipiens larvae were collected from three filariasis-endemic areas of Egypt and reared to adults for subsequent production and testing of F1 generation larvae and adults. Bioassays were performed using World Health Organization (WHO) methods with the diagnostic doses of 6 organophosphate insecticides for larvae and 1 organochlorine (OC), 4 pyrethroid, 2 organophosphate, and 2 carbamate insecticides for adults. Microplate assays were performed to measure levels of beta esterase, acetylcholinesterase, insensitive acetylcholinesterase, oxidases, and glutathione-S-transferase enzymes. Larval bioassay results showed clear indications of resistance to organophosphate insecticides. Adult bioassays also showed widespread, significant resistance to many insecticides from all four classes, including the OC, DDT. The Qalubiya larval population was susceptible only to malathion, whereas Sharkiya larvae were susceptible to malathion, temephos, and chlorpyrifos. On the other hand, larval specimens from Assiut were resistant to all insecticides tested. Larval bioassay results were supported by those of microplate assays in showing elevated levels of glutathione S-transferase in populations from all three areas. In general, microplate results confirmed patterns of resistance observed using bioassays, and mechanisms of resistance were evident for all three areas sampled. Mechanisms of resistance are discussed in relation to microplate and bioassay results for the areas sampled and pesticides used.
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