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Susceptibility of the Sandfly Phlebotomus Argentipes

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Susceptibility of the Sandfly Phlebotomus Argentipes Jpn. J. Infect. Dis., 68, 33–37, 2015 Original Article Susceptibility of the Sandfly Phlebotomus argentipes Annandale and Brunetti (Diptera: Psychodidae) to Insecticides in Endemic Areas of Visceral Leishmaniasis in Bihar, India Ram Singh1* andPramodKumar2 1National Centre for Disease Control, Patna branch, Bihar; and 2National Centre for Disease Control, Delhi, India SUMMARY: We present the results of susceptibility tests conducted on the sandfly Phlebotomus argen- tipes, the vector of visceral leishmaniasis in India. Adult P. argentipes insects were collected from 42 vil- lages in 6 districts of the state of Bihar, India, as follows: Patna, Vaishali, Muzaffarpur, Samastipur, Sheohar, and Sitamarhi. These adult insects were exposed to 4z DDT-, 5z malathion-, and 0.05z deltamethrin-impregnated papers using a WHO test kit by following the standard procedures. In 16 (38.1z) of 42 villages surveyed, the P. argentipes populations developed resistance to DDT. Susceptibil- ity tests using the organophosphate malathion in 22 villages revealed that in 1 (4.5z) village, the species developed resistance to this insecticide. P. argentipes was, however, highly susceptible to the synthetic pyrethroid deltamethrin. For long-term vector control of P. argentipes, it will be necessary to overcome the threat of insecticide resistance in this species. susceptibility test in this region (10). DDT resistance in INTRODUCTION P. argentipes populations was reported from some kala- Visceral leishmaniasis (VL) is a major public health azar-endemic areas in Bihar (11,12). However, informa- problem in India. Bangladesh, Nepal, and India signed tion on the status of insecticide resistance in the sandfly a memorandum of understanding and set an elimination is limited. Considering that the development of target of less than 1 case in 10,000 people by 2015 (1,2). resistance was reported in some places after prolonged The disease is transmitted by the bite of infected female use of DDT during IRS in endemic areas of Bihar, the sandflies, (Phlebotomus argentipes Annandale and present study was planned to investigate the susceptibil- Brunetti [Diptera: Psychodidae]), a well-known vector ity of P. argentipes in larger areas. The study was con- of VL called as kala-azar in India (3). P. argentipes is in ducted in highly endemic areas of VL to generate data general zoophilic but is also an opportunistic an- on the pattern of the susceptibility of P. argentipes to thropophile in nature (4). It prefers to breed in the soil DDT. This study also provided an opportunity to identi- of cattle shelters (5). The resting locations of adults are fy a potential insecticide that could be used as an alter- inside the cracks and crevices in the walls of human nate vector control strategy for kala-azar in the state of dwellings and cattle shelters (6), which are the target Bihar. Therefore, the organophosphate malathion and sites for the indoor residual spraying (IRS) of insecti- the synthetic pyrethroid deltamethrin were also tested, cides. Kala-azar was believed to have disappeared from as these are being widely used during IRS for the endemic areas during 1960–1970 as a collateral benefit malaria vector control program in India. of spraying DDT for malaria vector control under the National Malaria Control Program 1953 and National MATERIALS AND METHODS Malaria Eradication Program 1958 (7). It was a com- mon belief that the populations of P. argentipes were The study was conducted in 42 villages in 6 districts of effectively suppressed because of their higher level of Bihar, India. Among these villages, Hulashchak was susceptibility to DDT (8). Withdrawal of DDT spraying selected as the control village, as only one round of after 1962 by the National Malaria Eradication Pro- DDT spraying was performed in 1996 in 30 years. All gram was assumed to have resulted in increased vector the other villages were sprayed with DDT as part of the populations and consequently the resurgence of kala- kala-azar control program in the state of Bihar. The 6 azar during the mid-70s in the state of Bihar in India study districts of the state, Patna (25°25′N, 85°10′E), (8,9). Vaishali (25°30′N, 84°85′E), Muzaffarpur (26°23′N, Prior to 1979, P. argentipes wasconsideredtobe 85°45′E), Samastipur (25°5′N, 85°5′E), Sheohar highly susceptible to the insecticide of DDT in the WHO (26°2′N, 85°03′E), and Sitamarhi (26°49′N, 85°05′E), were selected based on their high endemicity of VL. Adult sandflies were collected from human dwellings Received August 14, 2013. Accepted May 30, 2014. and cattle shelters in each village in the early morning J-STAGE Advance Publication November 25, 2014. hours (0500–0700) with the help of a torch light and an DOI: 10.7883/yoken.JJID.2013.262 aspirator tube between March 2004 and November *Corresponding author: Mailing address: National Centre 2007. A total of 42 villages were evaluated for suscepti- for Disease Control, Patna branch, Bihar, India. Tel and bility to DDT, and because of the nonavailability of Fax: ＋916122572849, E-mail: ramsingh48＠hotmail.com sufficient numbers of sandflies for testing, malathion 33 Table 1. Susceptibility status of P. argentipes to 4z DDT in the WHO tube test 4z DDT at 1 h exposure Control District Village No. exposed z mortality No. exposed z mortality Corrected mortality Result1) Patna Hulashchak 90 100 30 0 100 S Mahinama Bazar 60 100 20 0 100 S Shanti nagar 60 96.7 15 0 96.7 T Janipur 60 100 15 0 100 S Dariyapur 60 100 20 0 100 S Gulmahia 60 66.7 18 0 66.7 R Gulmahia bagh 86 74.4 20 0 74.4 R Poonadih 60 90 16 0 90 T Naya Tola 60 80 20 0 80 T Sarayia 8095300 95 T Bakhara Chakia 45 95.5 15 0 95.6 T BhagahaTola 60 100 20 0 100 S Akopur 60 100 20 0 100 S Pulia Tola 60 100 20 5 100 S Majhaulia 52 98.1 20 0 98.1 S Vaishali Daulatpur Chandi 45 77.8 25 8 75.9 R MilePakari 60 53.3 15 0 53.3 R Rahimapur Pethia 60 78.3 15 6.7 76.7 R Kuwari bari Tola 48 68.8 16 6.3 66.5 R Muneem Chak 60 70 20 0 70 R Kanchanpur 60 95 15 0 95 T Jamalpur 60 93.3 20 0 93.3 T Wazitpur 48 97.9 20 0 97.9 T Kuwaribari Tola 60 73.3 18 0 73.3 R Islampur 60 63.3 25 8 60.1 R Chainpuramile pakari 60 85 18 5.6 84.1 T Muzaffarpur Gopnathpur Dokra 45 84.4 15 0 84.4 T GhandigramThuhma 50 78 15 6.7 76.4 R Chandra Hati 60 83.3 15 6.7 82.1 T Vijay Chhapara 45 93.3 16 6.3 92.9 T Pana pur 45 95.5 20 10 95 T Tarma Dushal Tola 45 95.6 18 0 95.6 T Akhatiyarpur 45 91.1 16 0 91.1 T Motipur 80 100 30 0 100 S Makshodpur 80 100 30 0 100 S Samastipur Jhhaypura 60 43.3 15 0 43.3 R Chandchouh 60 48.3 15 0 48.3 R Gangapur 55 54.5 15 0 54.5 R Bangra 50 64 15 6.7 61.4 R Mubarkpur 45 55.6 15 0 55.6 R Sheohar Rasheedpur 60 98.3 20 0 98.3 S Sitamarhi Ganeshpur 70 72.9 15 0 72.9 R 1): S, susceptible; T, tolerance; R, resistance. and deltamethrin susceptibility tests were performed sliding unit. The exposure tubes marked with red dots only in 22 and 20 villages, respectively. were lined with insecticide-impregnated paper and 1 The collected sandflies were brought to the laboratory control with carrier oil. All P. argentipes specimens in and released into a cage for acclimatization for 1 h ex- the holding tubes were transferred to the exposure tube clude insects that died of transportation stress. P. argen- using the sliding unit, and insects were kept in the ex- tipes specimens that were retained were used for suscep- posure tube for 1 h. The specimens were then trans- tibility tests, whereas other species of sandflies were ferred to a holding tube containing a 10z glucose removed. Papers impregnated with 4z DDT, 5z solution-soaked cotton pad. Mortality was recorded malathion, and 0.05z deltamethrin were used in sus- after 24 h of exposure. Abbot's formula was used to ceptibility tests by following the standard WHO suscep- correct the mortality rate if the mortality rate in the con- tibility test methods (13,14). trol was 5–20z ([(ztest mortality － zcontrol mortali- Blood-fed and active half-gravid females of P. argen- ty) × 100]/[100 － zcontrol mortality]). The tempera- tipes were used for the susceptibility test. Batches of ture and relative humidity were maintained at 25 ± 29C 15–25 sandflies were put in each holding tube with the and 75–80z, respectively. Three or 4 replicates were help of an aspirator tube through the filling hole in the performed for each insecticide in each village. The sus- 34 Insecticides Resistance in Sandfly P. argentipes Table 2. Susceptibility status of P. argentipes to 5z malathion in the WHO tube test 5z malathion at 1 h exposure Control District Village No. exposed z mortality No. exposed z mortality Corrected mortality Result1) Patna Dariyapur 45 100 20 0 100 S Gulmahiya 45 100 20 0 100 S Gulmahiya bagh 45 100 15 0 100 S Poonadih 45 100 20 0 100 S Bakharachakiya 45 100 15 0 100 S Akopur 45 100 16 0 100 S Vaishali Rahimapur Pethia 60 96.7 20 10 96.7 T Bidupur 45 100 16 0 100 S Muneemchak 45 100 18 0 100 S Muzaffarpur Gopnathpur Dokra 45 95.6 16 0 95.6 T Gandhigram Thumha 45 100 15 0 100 S Chandrahati 60 100 20 0 100 S Pana pur 45 100 15 0 100 S Tarma Dushal Tola 45 100 15 0 100 S Akhatiyarpur 45 100 15 6.7 100 S Vijayee Chhapara 45 100 15 0 100 S Samastipur Jhhaipura 45 84.4 15 0 84.4 T Chandchouh 40 87.5 15 0 87.5 T Gangapur 50 74 15 0 74 R Bangra 45 88.9 18 0 88.9 T Mubarakpur 45 95.6 16 0 95.6 T Sarayia 80 100 20 5 100 S 1): see Table 1 footnote.
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