Shri Prakash

Temephos-induced resistance in Aedes aegypti and its cross-resistance studies to certain insecticides from India

A temephos-induced resistance in Aedes aegypti that was developed for 24 generations exhibited 20.3-fold resistance as compared to susceptible strain. The diagnostic dose of temephos 0.02xa0mg/l exhibited gradual decrease in larval mortality with the progression of generations. An operational dose (1xa0mg/l) exhibited the LT50 value of 41.42xa0min in the susceptible population, whereas the value of the resistant population increased to 72.62, 108.86, 122.34, 182.03, 244.82 and 304.86xa0min in the fourth, eighth, 12th, 16th, 20th and 24th generation, respectively. The study carried out showed 120xa0min as the cut-off limit for differentiation between susceptible and resistant A. aegypti. Cross-resistance studies showed a varied degree of cross-resistance to fenthion, chlorpyrifos, malathion and DDT, whereas comparatively higher cross-resistance was observed to chlorpyrifos. Study on diagnostic doses of insecticides to A. aegypti females indicated a gradual decrease in adult mortality at every eighth generation as compared to susceptible population when exposed to malathion 0.8%, fenthion 0.25%, DDT 4%, permethrin 0.25% and temephos 6.5%, whereas little or no change in mortality when exposed to λ-cyhalothrin 0.03% and propoxur 0.1%, was observed. Thus, the expression of larval resistance was observed in adult stages also.

Electroantennogram, flight orientation, and oviposition responses of Aedes aegypti to the oviposition pheromone n -heneicosane

Oviposition pheromones specifically influence the females of many insects to lay eggs in the sites resulting in more egg deposition. A previous report describes the principal role of n-heneicosane (C21) identified and characterized from the larval cuticle of Aedes aegypti (L.) in attracting the gravid mosquitoes to oviposit in treated substrates among other chemical components. However, the means by which this compound is perceived by the females for oviposition has not been reported. In this study, we have recorded the peripheral olfactory responses from the antenna of Ae. aegypti from 10−7xa0g to 10−3xa0g doses of n-heneicosane. The EAG response of female mosquitoes increased in a dose-dependent manner with increasing stimulus strength. In the orientation assay using Y-maze olfactometer, female mosquitoes were attracted to the odor plume of 10−6xa0g and 10−5xa0g dose, while the higher dose of 10−3xa0g plume enforced repellency to gravid mosquitoes. The response to oviposition substrates by gravid Ae. aegypti females differed across the range of concentrations of n-heneicosane under multiple choice conditions, larger number of eggs were deposited in 10xa0ppm (10xa0mg/l) solutions compared to lower and higher concentrations indicating 10xa0ppm was most attractive. Application of n-heneicosane at 10xa0ppm in breeding habitats will be a useful method to attract the gravid mosquitoes using ovitraps for surveillance and monitoring. The possible use of this compound in monitoring of mosquito population in endemic areas in relevance to integrated vector management strategies is discussed in detail.

Electroantennogram and Behavioral Responses of Culex quinquefasciatus (Diptera: Culicidae) Females to Chemicals Found in Human Skin Emanations

Abstract The southern house mosquito, Culex quinquefasciatus Say (Diptera: Culicidae), is closely associated with humans and is a vector of filariasis. Use of semiochemicals for control of mosquitoes is a novel and potentially ecofriendly control approach. Human skin emanations are known to attract mosquitoes. In the current study, olfactory and behavioral responses of host-seeking female Cx. quinquefasciatus to various chemical components of human skin emanations (carboxylic acids, alcohols, and aldehydes) were evaluated separately at three doses (0.01, 0.1, and 1 μg) for electroantennogram (EAG) and three doses (0.1, 1, and 10 μg) for behavioral assay. Results of EAG studies indicated that all carboxylic acids elicited significant olfactory responses except tetradecanoic acid (C14) and octadecanoic acid (C18). In particular, hexanoic acid (C6) elicited a maximum, eight-fold olfactory response compared with the solvent control. Ethylene glycol and benzyl alcohol exhibited significant EAG and behavioral responses, whereas among aldehydes, nonanal elicited high EAG and behavioral responses, but only at all three doses tested compared with control. Some carboxylic acids elicited significant behavioral responses, attracting ≈75% of females tested toward chemical stimuli in a Y-tube olfactometer.

Highly Sensitive Amperometric Immunosensor for Detection of Plasmodium falciparum Histidine-Rich Protein 2 in Serum of Humans with Malaria: Comparison with a Commercial Kit

ABSTRACT A disposable amperometric immunosensor was developed for the detection of Plasmodium falciparum histidine-rich protein 2 (PfHRP-2) in the sera of humans with P. falciparum malaria. For this purpose, disposable screen-printed electrodes (SPEs) were modified with multiwall carbon nanotubes (MWCNTs) and Au nanoparticles. The electrodes were characterized by cyclic voltammetry, scanning electron microscopy, and Raman spectroscopy. In order to study the immunosensing performances of modified electrodes, a rabbit anti-PfHRP-2 antibody (as the capturing antibody) was first immobilized on an electrode. Further, the electrode was exposed to a mouse anti-PfHRP-2 antibody from a serum sample (as the revealing antibody), followed by a rabbit anti-mouse immunoglobulin G-alkaline phosphatase conjugate. The immunosensing experiments were performed on bare SPEs, MWCNT-modified SPEs, and Au nanoparticle- and MWCNT-modified SPEs (Nano-Au/MWCNT/SPEs) for the amperometric detection of PfHRP-2 in a solution of 0.1 M diethanolamine buffer, pH 9.8, by applying a potential of 450 mV at the working electrode. Nano-Au/MWCNT/SPEs yielded the highest-level immunosensing performance among the electrodes, with a detection limit of 8 ng/ml. The analytical results of immunosensing experiments with human serum samples were compared with the results of a commercial Paracheck Pf test, as well as the results of microscopy. The specificities, sensitivities, and positive and negative predictive values of the Paracheck Pf and amperometric immunosensors were calculated by taking the microscopy results as the “gold standard.” The Paracheck Pf kit exhibited a sensitivity of 79% (detecting 34 of 43 positive samples; 95% confidence interval [CI], 75 to 86%) and a specificity of 81% (correctly identifying 57 of 70 negative samples; 95% CI, 76 to 92%), whereas the developed amperometric immunosensor showed a sensitivity of 96% (detecting 41 of 43 positive samples; 95% CI, 93 to 98%) and a specificity of 94% (correctly identifying 66 of 70 negative samples; 95% CI, 92 to 99%). The developed method is more sensitive and specific than the Paracheck Pf kit.

Oviposition responses of Aedes aegypti and Aedes albopictus to certain fatty acid esters

Laboratory studies were carried out to observe the oviposition responses of Aedes aegypti (L.) and Aedes albopictus (Skuse) to several C21 fatty acid esters. The oviposition activity of these dengue and chikungunya vectors to the long-chain fatty acid esters of C21 length have not been reported earlier. From the multiple choice experiments on oviposition activity in standard mosquito cages, it was observed that compounds hexadecyl pentanoate, tetradecyl heptanoate and tridecyl octanoate presented significant oviposition repellent activity against the two mosquito species, while one compound propyl octadecanoate was found to attract A. aegypti to the treated oviposition substrate at 1- and 10-ppm concentrations. The possible utilization of these esters in integrated vector management is discussed.

Comparative activity of three repellents against the ticks Rhipicephalus sanguineus and Argas persicus

Abstract. The residual repellent activity of N, N‐diethylphenyl‐acetamide (DEPA), N, N‐diethyl‐3–methyl benzamide (DEET) and dimethylphthalate (DMP) against the hard tick Rhipicephalus sanguineus (Latreille) and the soft tick Argus persicus (Oken) from topical application on rabbits and hens respectively in the laboratory have been studied. It was found that a 25% concentration of DEET gave strong repellency (>90%) against larvae and nymphs of R. sanguineus for 15 and 7 days respectively and against A.persicus larvae and nymphs >90% for 11 and 5 days respectively. Strong repellency (>90%) against adults of the hard and soft ticks was observed for 9 and 7 days respectively with 40% DEET. DEPA at 25% concentration showed strong repellency (>90%) against larvae and nymphs of R.sanguineus and A.persicus for 11 and 5 days respectively. Adults of hard and soft ticks were strongly repelled (>90%) for 9 and 5 days respectively by 40% DEPA. DMP was found to be the least effective against all stages of both species of tick.

Resistance Status of the Malaria Vector Mosquitoes, Anopheles stephensi and Anopheles subpictus Towards Adulticides and Larvicides in Arid and Semi-Arid Areas of India

Abstract Susceptibility studies of malaria vectors Anopheles stephensi Liston (Diptera: Culicidae) and An. subpictus Grassi collected during 2004–2007 from various locations of Arid and Semi-Arid Zone of India were conducted by adulticide bioassay of DDT, malathion, deltamethrin and larvicide bioassay of fenthion, temephos, chlorpyriphos and malathion using diagnostic doses. Both species from all locations exhibited variable resistance to DDT and malathion from majority of location. Adults of both the species were susceptible to Deltamethrin. Larvae of both the Anopheline species showed some evidence of resistance to chlorpyriphos followed by fenthion whereas susceptible to temephos and malathion.

Susceptibility of immature stages of Aedes (Stegomyia) aegypti; vector of dengue and chikungunya to insecticides from India

Susceptibility of Aedes aegypti to some insecticides in different geographic areas was conducted during dengue and chikungunya outbreak. At present, the only method of preventing dengue and chikungunya is to control the vector, which is the weakest link in vector-borne diseases. In our study, the susceptibility of A. aegypti collected from urban areas of Delhi, Mumbai, Jodhpur, Chennai and Coimbatore was evaluated against temephos, fenthion, malathion and DDT. The A. aegypti from different locations exhibited 0.33–7.11, 0.36–3.00, 0.65–2.84 and 2.16–20.8 fold more lethal concentration value of 50% (LC50) to temephos, fenthion, malathion and DDT, respectively, compared to susceptible reference strain. The result reveals that A. aegypti from various locations studied are still susceptible to temephos, fenthion and malathion, whereas low level of DDT resistance was noticed in field-collected A. aegypti. Amongst the insecticides tested, temephos was found to be relatively more effective in controlling A. aegypti, followed by fenthion, malathion and DDT.

Midgut Microbial Community of Culex quinquefasciatus Mosquito Populations from India

The mosquito Culex quinquefasciatus is a ubiquitous species that serves as a major vector for west nile virus and lymphatic filariasis. Ingestion of bloodmeal by females triggers a series of physiological processes in the midgut and also exposes them to infection by these pathogens. The bacteria normally harbored in the midgut are known to influence physiology and can also alter the response to various pathogens. The midgut bacteria in female Cx. quinquefasciatus mosquitoes collected over a large geographical area from India was studied. Examination of 16S ribosomal DNA amplicons from culturable microflora revealed the presence of 83 bacterial species belonging to 31 bacterial genera. All of these species belong to three phyla i.e. Proteobacteria, Firmicutes and Actinobacteria. Phylum Proteobacteria was the most dominant phylum (37 species), followed by Firmicutes (33 species) and Actinobacteria (13 species). Phylum Proteobacteria, was dominated by members of γ-proteobacteria class. The genus Staphylococcus was the largest genus represented by 11 species whereas Enterobacter was the most prevalent genus and recovered from all the field stations except Leh. Highest bacterial prevalence was observed from Bhuj (22 species) followed by Nagrota (18 species), Masimpur (18 species) and Hathigarh (16 species). Whereas, least species were observed from Leh (8 species). It has been observed that individual mosquito harbor extremely diverse gut bacteria and have very small overlap bacterial taxa in their gut. This variation in midgut microbiota may be one of the factors responsible for variation in disease transmission rates or vector competence within mosquito population. The present data strongly encourage further investigations to verify the potential role of the detected bacteria in mosquito for the transmission of lymphatic filariasis and west nile virus. To the best of our knowledge this is the first study on midgut microbiota of wild Cx. quinquefasciatus from over a large geographical area.

Scanning electron microscopic studies on egg surface morphology and morphometrics of Culex tritaeniorhynchus and Culex quinquefasciatus (Diptera: Culicidae)

Culex tritaeniorhynchus Giles, 1901 and Culex quinquefasciatus Say, 1823 is an important vector of Japanese encephalitis and Bancroftian filariasis, respectively in India and South East Asian countries. In this paper, we are describing the surface morphology and morphometrics of the egg of C. tritaeniorhynchus in comparison with C. quinquefasciatus for the first time. The results indicated that eggs of both the species appears to be similar to great extent in surface morphology, however, morphometrics provide 56.81% demarking attributes out of 44 attributes at various significant levels (pu2009<u20090.05–0.001), i.e., egg length, width and ratio of length/width, attributes of micropylar apparatus including corolla, disc, mound, tubercles size, and length of tubercular rows in micropylar region, size and density of tubercles, exochorionic pore in conical-shaped regions of eggs, and size of tubercular wheel units. Structurally, the additional presence of large tubercles strengthens the micropylar region to bear various collapsing forces in these species.