Susceptible Laboratory Strain Research Articles

Impact of deltamethrin-resistance in Aedes albopictus on its fitness cost and vector competence

Aedes albopictus is one of the most invasive species in the world as well as the important vector for mosquito-borne diseases such as dengue fever, chikungunya fever and zika virus disease. Chemical control of mosquitoes is an effective method to control mosquito-borne diseases, however, the wide and improper application of insecticides for vector control has led to serious resistance problems. At present, there have been many reports on the resistance to pyrethroid insecticides in vector mosquitoes including deltamethrin to Aedes albopictus. However, the fitness cost and vector competence of deltamethrin resistant Aedes albopictus remain unknown. To understand the impact of insecticide resistant mosquito is of great significance for the prevention and control mosquitoes and mosquito-borne diseases. A laboratory resistant strain (Lab-R) of Aedes albopictus was established by deltamethrin insecticide selecting from the laboratory susceptible strain (Lab-S). The life table between the two strains were comparatively analyzed. The average development time of Lab-R and Lab-S in larvae was 9.7 days and 8.2 days (P < 0.005), and in pupae was 2.0 days and 1.8 days respectively (P > 0.05), indicating that deltamethrin resistance prolongs the larval development time of resistant mosquitoes. The average survival time of resistant adults was significantly shorter than that of susceptible adults, while the body weight of resistant female adults was significantly higher than that of the susceptible females. We also compared the vector competence for dengue virus type-2 (DENV-2) between the two strains via RT-qPCR. Considering the results of infection rate (IR) and virus load, there was no difference between the two strains during the early period of infection (4, 7, 10 day post infection (dpi)). However, in the later period of infection (14 dpi), IR and virus load in heads, salivary glands and ovaries of the resistant mosquitoes were significantly lower than those of the susceptible strain (IR of heads, salivary glands and ovaries: P < 0.05; virus load in heads and salivary glands: P < 0.05; virus load in ovaries: P < 0.001). And then, fourteen days after the DENV-2-infectious blood meal, females of the susceptible and resistant strains were allow to bite 5-day-old suckling mice. Both stains of mosquito can transmit DENV-2 to mice, but the onset of viremia was later in the mice biting by resistant group as well as lower virus copies in serum and brains, suggesting that the horizontal transmission of the resistant strain is lower than the susceptible strain. Meanwhile, we also detected IR of egg pools of the two strains on 14 dpi and found that the resistant strain were less capable of vertical transmission than susceptible mosquitoes. In addition, the average survival time of the resistant females infected with DENV-2 was 16 days, which was the shortest among the four groups of female mosquitoes, suggesting that deltamethrin resistance would shorten the life span of female Aedes albopictus infected with DENV-2. As Aedes albopictus developing high resistance to deltamethrin, the resistance prolonged the growth and development of larvae, shorten the life span of adults, as well as reduced the vector competence of resistant Aedes albopictus for DENV-2. It can be concluded that the resistance to deltamethrin in Aedes albopictus is a double-edged sword, which not only endow the mosquito survive under the pressure of insecticide, but also increase the fitness cost and decrease its vector competence. However, Aedes albopictus resistant to deltamethrin can still complete the external incubation period and transmit dengue virus, which remains a potential vector for dengue virus transmission and becomes a threat to public health. Therefore, we should pay high attention for the problem of insecticide resistance so that to better prevent and control mosquito-borne diseases.

Evaluation of triflumuron and pyriproxyfen as alternative candidates to control house fly, Musca domestica L. (Diptera: Muscidae), in Riyadh city, Saudi Arabia.

This study was conducted to determine the susceptibility and resistance of some house fly strains of Musca domestica L. to the insect growth regulator insecticides triflumuron and pyriproxyfen in some locations in Riyadh city. Field-collected strains of M. domestica L. from five sites in Riyadh city that represented five slaughterhouse sites where flies spread significantly were tested against triflumuron and pyriproxyfen. Triflumuron LC50 values for the five collected strains ranged from 2.6 to 5.5 ppm, and the resistance factors (RFs) ranged from 13-fold to 27-fold that of the susceptible laboratory strain. Pyriproxyfen LC50 values for the field strains ranged from 0.9 to 1.8 ppm with RFs of 3-fold to 5-fold. These results indicate that pyriproxyfen is an effective insecticide to control house flies and should be used in rotation with other insecticides in the control programs applied by Riyadh municipality.

Перспективы применения новых действующих веществ для борьбы с комнатными мухами

The toxicity of three new insecticides indoxacarb, chlorfenapyr and fluralaner for house flies of the susceptible laboratory strain S-NIID has been studied. A comparison is made of their insecticidal properties in the contact and intestinal routes of entry into the body. Sugar baits with this three insecticides are highly toxic to houseflies. Insecticidal baits containing 0.1–0.6 % a.i. of indoxacarb are highly effective against houseflies. Apparently, indoxacarb, chlorfenapyr, and fluralaner can be recommended for the introduction of insecticides into rotation schemes to combat winged flies, both in the form of concentrates used by spraying or insecticidal baits.

Insecticide resistance status of malaria vectors Anopheles gambiae (s.l.) of southwest Burkina Faso and residual efficacy of indoor residual spraying with microencapsulated pirimiphos-methyl insecticide

BackgroundThe rapid spread of insecticide resistance in malaria vectors and the rebound in malaria cases observed recently in some endemic areas underscore the urgent need to evaluate and deploy new effective control interventions. A randomized control trial (RCT) was conducted with the aim to investigate the benefit of deploying complementary strategies, including indoor residual spraying (IRS) with pirimiphos-methyl in addition to long-lasting insecticidal nets (LLINs) in Diébougou, southwest Burkina Faso.MethodsWe measured the susceptibility of the Anopheles gambiae (s.l.) population from Diébougou to conventional insecticides. We further monitored the efficacy and residual activity of pirimiphos-methyl on both cement and mud walls using a laboratory susceptible strain (Kisumu) and the local An. gambiae (s.l.) population.ResultsAn. gambiae (s.l.) from Diébougou was resistant to DDT, pyrethroids (deltamethrin, permethrin and alphacypermethrin) and bendiocarb but showed susceptibility to organophosphates (pirimiphos-methyl and chlorpyrimiphos-methyl). A mixed-effect generalized linear model predicted that pirimiphos-methyl applied on cement or mud walls was effective for 210 days against the laboratory susceptible strain and 247 days against the local population. The residual efficacy of pirimiphos-methyl against the local population on walls made of mud was similar to that of cement (OR = 0.792, [0.55–1.12], Tukey’s test p-value = 0.19).ConclusionsIf data on malaria transmission and malaria cases (as measured trough the RCT) are consistent with data on residual activity of pirimiphos-methyl regardless of the type of wall, one round of IRS with pirimiphos-methyl would have the potential to control malaria in a context of multi-resistant An. gambiae (s.l.) for at least 7 months.Graphical

Larvicidal potential of two silver nano-particles (Moringa oleifera and Ficus exasperata) against laboratory and field strains of Anopheles gambiae (Diptera: Culicidae) in Lagos, Nigeria

BackgroundThe interest in larvicides of plant origin is generally renewed in vector control because of their safety compared to synthetic larvicides. However, there are concerns about the relative safety dose of these phytochemicals on non-target organisms which led to the development of plant derived nanoparticles. In this study, we examined the bioefficacy of low doses of two green synthesized nanoparticles on immature stages of Anopheles mosquitoes in Nigeria. Aqueous plants (Moringa oleifera and Ficus exasperata) extracts were used in the biosynthesis. The prepared Ag-NPs were characterizations using Fourier-transform infrared spectroscopy (FT-IR), UV-Vis spectroscopy, and scanning electron microscopy (SEM). Third and early fourth instars of known susceptible laboratory strains of Anopheles gambiae s.s. (KISUMU strains) and pyrethroid resistant field strain of An. gambiae were exposed to serial dilutions of 0.25, 0.5, 0.75, 1.0 and 2.5 ppm of each phyto nanoparticles. Moribund and dead larvae were observed after 24 and 48 h post exposure, and the results were analysed with descriptive statistics.ResultsWith the laboratory mosquitoes, Moringa oleifera AgNP effected high mortalities of 88–100% (LC50 = 0.39 ppm; LC95 = 0.62 ppm) at 24 h post exposure except at the lowest concentration, while Ficus exasperate AgNP induced a 32–100% mortality (LC50 = 0.51 ppm; LC95 = 1.15 ppm) except at the lowest concentration. In the field populations, mortality in Moringa oleifera and Ficus exasperata was 23–93% (LC50 = 0.65 ppm; LC95 = 2.28 ppm) and 37–50% (LC50 = 1.51 ppm; LC95 = 391.64 ppm) respectively. There was no significant difference in mortality values between the laboratory and field strains (P < 0.05) at both 24 and 48 h post exposure times.ConclusionsOverall, the study demonstrates the bioefficacy and potential use of green synthesized nanoparticles, at very low concentrations for the control of Anopheles larvae even in areas where resistance to the current chemical insecticides have been reported.

Determination of insecticide resistance in Bemisia tabaci (Hemiptera: Aleyrodidae) populations from Punjab, Pakistan

The whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is one of the most important crop pest among the 100 worst invasive species which threatens agriculture worldwide and poses a serious threat to global food security. The control of B. tabaci depends on the insecticide applications in Pakistan, therefore, monitoring of insecticide resistance is essential for achieving control and for managing resistance. In current study, the field populations of B. tabaci from three crops (cotton, okra and tomato) were collected in five districts (Bahawalpur, Lodhran, Multan, Vehari and Faisalabad) of Punjab, Pakistan in 2017, 2018 and 2019. A leaf- dip bioassay was used to determine the resistance of these populations to five insecticides (Diafenthiuron, spirotetramat, deltamethrin, abamectin and nitenpyram). A very low to low level of resistance to nitenpyram, spirotetramat, and abamectin, whereas moderate to high level of resistance to diafenthiuron and deltamethrin were detected in all populations of B. tabaci compared with the laboratory susceptible strain. The resistance ratios ranged 16.37 to 26.07-fold for diafenthiuron, 52.76 to 73.85-fold for deltamethrin, 3.85 to 7.79-fold for nitenpyram, 3.62 to 13.75-fold for spirotetramat and 5.19 to 9.09-fold for abamectin. The presented results will be helpful to make decision on the proper usage of insecticides against whitefly control and to reduce the development of insecticide resistance.

Geographic Variation in Dominance of Spinosad Resistance in Colorado Potato Beetles (Coleoptera: Chrysomelidae).

Eastern New York State is frequently the site of Colorado potato beetle (Leptinotarsa decemlineata, Say) populations with the highest observed levels of insecticide resistance to a range of active ingredients. The dominance of a resistant phenotype will affect its rate of increase and the potential for management. On organic farms on Long Island, L. decemlineata evolved high levels of resistance to spinosad in a short period of time and that resistance has spread across the eastern part of the Island. Resistance has also emerged in other parts of the country as well. To clarify the level of dominance or recessiveness of spinosad resistance in different parts of the United States and how resistance differs in separate beetle populations, we sampled in 2010 beetle populations from Maine, Michigan, and Long Island. In addition, a highly resistant Long Island population was assessed in 2012. All populations were hybridized with a laboratory-susceptible strain to determine dominance. None of the populations sampled in 2010 were significantly different from additive resistance, but the Long Island population sampled in 2012 was not significantly different from fully recessive. Recessive inheritance of high-level resistance may help manage its increase.

Fitness costs associated with chlorantraniliprole resistance in Spodoptera exigua (Lepidoptera: Noctuidae).

The field population of Spodoptera exigua, an intermittently occurring polyphagous pest, has developed resistance to chlorantraniliprole, while whether or not such resistance carries fitness costs remains poorly understood. Here we selected six generations of the Leshan population (LS-P) by two-way selecting method, and obtained a highly resistant strain (CH-RE) and resistant degeneration strain (CH-SE) sharing a similar genetic background. After that fitness costs were evaluated by comparing the life history characteristics of CH-RE, CH-SE and the laboratory susceptible strain (SE-Lab) via the age-stage two-sex life table method. The resistance ratio of CH-RE and CH-SE were 226.69-fold and 3.72-fold, respectively, and the estimated realized heritability (h2 ) of CH-RE was 0.058. Compared with CH-SE, the duration of pre-adult, the longevity of adult, adult preoviposition period (APOP) and average generation time (T) of CH-RE had significantly increased, but the oviposition days, average fecundity, intrinsic growth rate (r), weekly growth rate (λ) and reproductive rate (R0 ) decreased significantly. Moreover, the relative fitness of CH-RE was 0.25, and showed fitness costs. Concurrently, the fecundity of CH-SE was slightly lower than SE-Lab, but there was no significant difference in r, λ and R0 , and the fitness (1) of CH-SE was similar to SE-Lab (1.02), which was no fitness cost. These findings represent that chlorantraniliprole resistance in S. exigua has a fitness cost, and the fitness cost will disappear with the recovery of sensitivity when the insecticide is stopped for field populations, supporting that such resistance would be managed by switching off the selection pressure with rotation with alternate insecticides. © 2020 Society of Chemical Industry.

Resistance status and detoxification enzyme activity in ten populations of Panonychus citri (Acari: Tetranychidae) from Turkey

Acaricide resistance monitoring in phytophagous mites is crucial for effective pest management. Citrus growers in Cukurova region, a region that comprised more than 75% of total citrus production in Turkey, complained that acaricides have become inefficient in controlling outbreaks of the citrus red mite, Panonychus citri McGregor (Acari: Tetranychidae). To assess whether such control failures were caused by acaricide resistance, we bio-assayed P. citri populations collected from nine commercial orchards and a susceptible laboratory strain. Based on the mode of action (MoA) of acaricides, protonymphs were bio-assayed with etoxazole, spirodiclofen and spirotetramat, whereas abamectin and pyridaben were used for adults. Additionally, we determined kinetic esterase and glutathione S-transferase (GST) enzyme activities using a micro plate reader. Based on LC50 values, the resistance ratios of the orchard populations compared to the laboratory population varied from 2.81 to 34.82-fold, 1.22–18.35-fold, 1.23–40.43-fold, 1.76–27.50-fold and 2.24–75.06-fold for abamectin, etoxazole, spirodiclofen, spirotetramat and pyridaben, respectively. Total esterase and GST activities of the populations were in the ranges of 15.86–43.28 and 2.07–5.29 mOD/min/mg protein, respectively. The highest enzyme activity levels were detected in the Yakapinar population which was among one of the most resistant populations to all tested acaricides. The activities of detoxification enzymes may play an important role in the metabolic breakdown of the acaricides in resistant P. citri populations. Rotation of acaricides with different modes of action is fundamental to prevent potential resistance problems.

Transcriptome Analysis of Pyrethroid-Resistant Chrysodeixis includens (Lepidoptera: Noctuidae) Reveals Overexpression of Metabolic Detoxification Genes.

Chrysodeixis includens (Walker, [1858]) is one of the most important defoliator of soybean in Brazil because of its extensive geographical distribution and high tolerance to insecticides compared with other species of caterpillars. Because of this, we conducted bioassays to evaluate the efficacy of pyrethroid λ-cyhalothrin on a C. includens resistant strain (MS) and a susceptible (LAB) laboratory strain. High throughput RNA sequencing (RNA-seq) of larval head and body tissues were performed to identify potential molecular mechanisms underlying pyrethroid resistance. Insecticide bioassays showed that MS larvae exhibit 28.9-fold resistance to pyrethroid λ-cyhalothrin relative to LAB larvae. RNA-seq identified evidence of metabolic resistance in the head and body tissues: 15 cytochrome P450 transcripts of Cyp6, Cyp9, Cyp4, Cyp304, Cyp307, Cyp337, Cyp321 families, 7 glutathione-S-transferase (Gst) genes, 7 α-esterase genes from intracellular and secreted catalytic classes, and 8 UDP-glucuronosyltransferase (Ugt) were overexpressed in MS as compared with LAB larvae. We also identified overexpression of GPCR genes (CiGPCR64-like and CiGPCRMth2) in the head tissue. To validate RNA-seq results, we performed RT-qPCR to assay selected metabolic genes and confirmed their expression profiles. Specifically, CiCYP9a101v1, CiCYP6ae149, CiCYP6ae106v2, CiGSTe13, CiCOE47, and CiUGT33F21 exhibited significant overexpression in resistant MS larvae. In summary, our findings detailed potential mechanisms of metabolic detoxification underlying pyrethroid resistance in C. includens.

Mechanisms for multiple resistances in field populations of rice stem borer, Chilo suppressalis (Lepidoptera: Crambidae) from Sichuan Province, China

Chilo suppressalis Walker (Lepidoptera: Crambidae) is a widely destructive pest occurring in rice, particularly in the rice-growing regions of Asia. In recent years, C. suppressalis has developed resistance to several insecticides because of the extensive use of insecticides. The resistance levels to four insecticides were determined among populations from different regions of Sichuan Province, China, using a drop-method bioassay. Based on LC50 values of a laboratory susceptible strain, all field populations showed moderate level of resistance to triazophos (23.9- to 83.5-fold) and were either susceptible or had a low level of resistance to abamectin (2.1- to 5.8-fold). All field-collected populations had a low or moderate level of resistance to chlorpyrifos (1.7- to 47.1-fold) and monosultap (2.7- to 13.5-fold). The synergism experiment indicated that the resistance of the XW19 to triazophos may be associated with cytochrome P450 monooxygenases (P450s), with the highest synergistic ratio (SR) of 3.05-fold and increased ratio (IR) of 2.28-fold for piperonylbutoxide (PBO). The P450 activity of the TJ19 population was the greatest among the six field populations. Moreover, the relative expression levels of four resistance-related P450 genes were detected with qRT-PCR, and the results indicated that CYP324A12, CYP321F3 and CYP9A68 were overexpressed in the resistant population, especially in the XW19 population (by 1.2-, 3.4 -, and 18.0-fold, respectively). In addition, the relative expression levels of CYP9A68 among the CZ19 and TJ19 populations were also enhanced 10.5- and 24.9-fold, respectively. These results suggested that CYP324A12, CYP321F3 and CYP9A68 may be related to the resistance development of C. suppressalis to triazophos.

Fitness cost of insecticide resistance on the life-traits of a Anopheles coluzzii population from the city of Yaoundé, Cameroon.

Background: Pyrethroid resistance is rapidly expanding in An. gambiae s.l. populations across Sub-Saharan Africa. Yet there is still not enough information on the fitness cost of insecticide resistance . In the present study, the fitness cost of insecticide resistance on Anopheles coluzzii population from the city of Yaoundé was investigated. Methods: A resistant An. coluzzii colony was established from field collected mosquitoes resistant to both DDT and pyrethroid and selected for 12 generations with deltamethrin 0.05%. The Ngousso laboratory susceptible strain was used as control. A total of 100 females of each strain were blood fed and allowed for individual eggs laying, and then different life traits parameters such as fecundity, fertility, larval development time, emergence rate and longevity were measured. The TaqMan assay was used to screen for the presence of the L1014F and L1014S kdr mutations. Results: Field collected mosquitoes from the F0 generation had a mortality rate of 2.05% for DDT, 34.16% for permethrin and 50.23% for deltamethrin. The mortality rate of the F12 generation was 30.48% for deltamethrin, 1.25% for permethrin and 0% for DDT. The number of eggs laid per female was lower in the resistant colony compared to the susceptible (p <0.0001). Insecticide resistant larvae were found with a significantly long larval development time (10.61±0.33 days) compare to susceptible (7.57±0.35 days). The number of emerging females was significantly high in the susceptible group compared to the resistant . The adults lifespan was also significantly high for susceptible (21.73±1.19 days) compared to resistant (14.63±0.68 days). Only the L1014F- kdr allele was detected in resistant population.. Conclusion: The study suggests that pyrethroid resistance is likely associated with a high fitness cost on An.coluzzii populations. The addition of new tools targeting specifically larval stages could improve malaria vectors control and insecticide resistance management.

Fitness cost of insecticide resistance on the life-traits of a Anopheles coluzzii population from the city of Yaoundé, Cameroon

Background: Pyrethroid resistance is rapidly expanding in An. gambiae s.l. populations across Sub-Saharan Africa. Yet there is still not enough information on the fitness cost of insecticide resistance . In the present study, the fitness cost of insecticide resistance on Anopheles coluzzii population from the city of Yaoundé was investigated. Methods: A resistant An. coluzzii colony was established from field collected mosquitoes resistant to both DDT and pyrethroid and selected for 12 generations with deltamethrin 0.05%. The Ngousso laboratory susceptible strain was used as control. A total of 100 females of each strain were blood fed and allowed for individual eggs laying, and then different life traits parameters such as fecundity, fertility, larval development time, emergence rate and longevity were measured. The TaqMan assay was used to screen for the presence of the L1014F and L1014S kdr mutations. Results: Field collected mosquitoes from the F0 generation had a mortality rate of 2.05% for DDT, 34.16% for permethrin and 50.23% for deltamethrin. The mortality rate of the F12 generation was 30.48% for deltamethrin, 1.25% for permethrin and 0% for DDT. The number of eggs laid per female was lower in the resistant colony compared to the susceptible (p <0.0001). Insecticide resistant larvae were found with a significantly long larval development time (10.61±0.33 days) compare to susceptible (7.57±0.35 days). The number of emerging females was significantly high in the susceptible group compared to the resistant . The adults lifespan was also significantly high for susceptible (21.73±1.19 days) compared to resistant (14.63±0.68 days). Only the L1014F- kdr allele was detected in resistant population.. Conclusion: The study suggests that pyrethroid resistance is likely associated with a high fitness cost on An.coluzzii populations. The addition of new tools targeting specifically larval stages could improve malaria vectors control and insecticide resistance management.

Repellency and Contact Irritancy Responses of Aedes aegypti (Diptera: Culicidae) Against Deltamethrin and Permethrin: A Cross-Regional Comparison.

Control strategies exploiting the innate response of mosquitoes to chemicals are urgently required to complement existing traditional approaches. We therefore examined the behavioral responses of 16 field strains of Aedes aegypti (L.) from two countries, to deltamethrin and permethrin by using an excito-repellency (ER) test system. The result demonstrated that the escape percentage of Ae. aegypti exposed to pyrethroids did not vary significantly between the two countries in both contact and noncontact treatment despite the differing epidemiological patterns. Deltamethrin (contact: 3.57 ± 2.06% to 31.20 ± 10.71%; noncontact: 1.67 ± 1.67% to 17.31 ± 14.85%) elicited relatively lower responses to field mosquitoes when compared with permethrin (contact: 16.15 ± 4.07% to 74.19 ± 4.69%; noncontact: 3.45 ± 2.00% to 41.59 ± 6.98%) in contact and noncontact treatments. Compared with field strains, the mean percentage of escaping laboratory susceptible strain individuals were significantly high after treatments (deltamethrin contact: 72.26 ± 6.95%, noncontact: 61.10 ± 12.31%; permethrin contact: 78.67 ± 9.67%, noncontact: 67.07 ± 7.02%) and the escaped individuals spent significantly shorter time escaping from the contact and noncontact chamber. The results indicated a significant effect of resistance ratio on mean escape percentage, but some strains varied idiosyncratically compared to the increase in insecticide resistance. The results also illustrated that the resistance ratio had a significant effect on the mortality in treatments. However, the mortality in field mosquitoes that prematurely escaped from the treated contact chamber or in mosquitoes that stayed up to the 30-min experimental period showed no significant difference.

Phosphine resistance in saw-toothed grain beetle, Oryzaephilus surinamensis in the United States

Phosphine (PH3) fumigation resulting in sub-lethal exposure has led to the development of phosphine resistance in many stored-product insect species worldwide and is a major challenge to the continued effective use of phosphine. In 2016 phosphine resistance was found in Tribolium castaneum (Herbst) and Plodia interpunctella (Hübner) collected from California dried fruit and nut processing facilities. Although Oryzaephilus surinamensis (L.) infests grain, dried fruit, and nuts in storage and processing facilities, phosphine resistance in this species has not been studied in the United States. In this study, the discriminating dose of phosphine for O. surinamensis eggs was estimated using a laboratory susceptible strain; it was found to be 28.4 ppm over a 72-h fumigation period (1 mg/L of phosphine = 714.18 ppm or 1 ppm = 0.0014 mg/L). Discriminating dose bioassays were used to determine phosphine resistance in both eggs and adults of 14 different populations collected from California and Oklahoma. Resistance to phosphine was detected in four out of 14 populations in adults and nine out of 14 populations in eggs and ranged from 2 to 100%. Phosphine percent survival values in both adults and eggs of three populations, namely, Box BR, Box BF, and OKWat were >90%. Lethal concentration values required to kill 99% of individuals in samples for adults of these three populations were predicted as 320.5, 290.7, and 263 ppm, respectively, and those for eggs were 1030.7, 1055.9, and 564.5 ppm, respectively, over a 72-h fumigation period. This study confirms that phosphine resistance is present in O. surinamensis in the United States.

New in vitro model to evaluate kinetics of antimycobacterial drug release from bioresorbable polymeric carriers

Sustained-release drugs against tuberculosis are a promising approach to therapy since they positively affect patient compliance with long regimens, especially when it comes to the multidrug-resistant form of the disease. Conventional UV-visible spectroscopy does not work well with multicomponential culture media used for growing M. tuberculosis. The aim of this study was to develop a method for evaluating the kinetics of anti-tuberculosis drug released from bioresorbable polymeric carriers suitable for screening a wide range of encapsulated prolonged-release drugs and identifying the best performing candidate. While studying the growth dynamics of the laboratory susceptible strain M. tuberculosis H37Rv in the presence of different levofloxacin concentrations (from 0.03 to 0.4 μg/ml), we developed a model, which is essentially a set of 2 parallel experiments evaluating the kinetics of drug release into the culture medium. The results of these 2 experiments conducted on 3 encapsulated forms of levofloxacin loaded onto bioresorbable polymeric PLGA carriers (particles sized 50 μm and 100 μm and the matrix) revealed that release kinetics of the drug largely depended on the type of polymeric carrier. The best encapsulation of the antibiotic and its gradual release into the culture medium was observed for the matrix. All experiments were run in 3 replicates. The obtained data were analyzed using descriptive statistics.

Development of resistance in stored grain pest, Trogoderma granarium (Everts) against deltamethrin and its effective control by synergistic toxicity of bifenthrin and chlorpyrifos

The current investigation was conducted to estimate the resistance level against deltamethrin in fourth and sixth instar larvae of Trogoderma granarium collected from wheat storage warehouses of Dera Ghazi Khan (DGK), Okara (OKR) and Gujranwala (GUW). Results showed that the fourth and sixth instar larvae of collected strains warehouses moderately resistant to deltamethrin compared with the same developmental stages of insecticide susceptible laboratory strain (LAB-S). Efficacies of bifenthrin, chlorpyrifos and their various combinations were analyzed against fourth and sixth instar larvae of different strains of pest populations to minimize the development of resistance against deltamethrin. LC50s of bifenthrin against fourth and sixth instar larvae of all the four populations were recorded at low concentrations compared to deltamethrin. The LC50 values of bifenthrin against fourth instar larvae of LAB-S, DGK, OKR and GUW populations were 359.32, 416.58, 524.01 and 598.91 ppm, respectively, while these values were 204.30, 262.98, 301.89 and 355.22 ppm against sixth instar larvae of those populations, respectively. Chlorpyrifos was found to be the most effective insecticide compared to bifenthrin and deltamethrin. The LC50 values of chlorpyrifos were 189.33, 305.37, 376.30 and 459.15 ppm against fourth instar larvae of LAB-S, DGK, OKR and GUW populations, respectively, whereas for sixth instar larvae those values were 98.24, 129.61, 163.43 and 230.74 ppm, respectively. Various combinations of bifenthrin and chlorpyrifos viz., 1:1, 2:1, 3:1, 1:2 and 1:3 were also tested. Based on the values of LC50 and relative toxic unit, 1:3 combination was observed as the most effective among all combinations. The LC50 values of fourth instar larvae of all the four populations against each insecticide and their combinations were recorded at high concentration compared to that of sixth instar larvae. Results of current investigation need to be considered in present stored grain insect pest control program of warehouses in Punjab1.

Fitness cost of insecticide resistance on the life-traits of a Anopheles coluzzii population from the city of Yaoundé, Cameroon

Background: Pyrethroid resistance is rapidly expanding in An. gambiae s.l. populations across Sub-Saharan Africa. Yet there is still not enough information on the fitness cost of insecticide resistance . In the present study, the fitness cost of insecticide resistance on Anopheles coluzzii population from the city of Yaoundé was investigated. Methods: A resistant An. coluzzii colony was established from field collected mosquitoes resistant to both DDT and pyrethroid and selected for 12 generations with deltamethrin 0.05%. The Ngousso laboratory susceptible strain was used as control. A total of 100 females of each strain were blood fed and allowed for individual eggs laying, and then different life traits parameters such as fecundity, fertility, larval development time, emergence rate and longevity were measured. The TaqMan assay was used to screen for the presence of the L1014F and L1014S kdr mutations. Results: Field collected mosquitoes from the F0 generation had a mortality rate of 2.05% for DDT, 34.16% for permethrin and 50.23% for deltamethrin. The mortality rate of the F12 generation was 30.48% for deltamethrin, 1.25% for permethrin and 0% for DDT. The number of eggs laid per female was lower in the resistant colony compared to the susceptible (p &lt;0.0001). Insecticide resistant larvae were found with a significantly long larval development time (10.61±0.33 days) compare to susceptible (7.57±0.35 days). The number of emerging females was significantly high in the susceptible group compared to the resistant . The adults lifespan was also significantly high for susceptible (21.73±1.19 days) compared to resistant (14.63±0.68 days). Only the L1014F-kdr allele was detected in resistant population.. Conclusion: The study suggests that pyrethroid resistance is likely associated with a high fitness cost on An.coluzzii populations. The addition of new tools targeting specifically larval stages could improve malaria vectors control and insecticide resistance management.

Toxicity evaluation of Spinosad against Malathion and deltamethrin resistant population of rusty grain beetle, Cryptolestes ferrugenius (Stephens) in Andhra Pradesh, India

A laboratory experiment was implemented in the Department of Entomology Agricultural College, Bapatla to know the resistance levels of malathion and deltamethrin against Rusty grain beetle, Cryptolestes ferrugenius (Stephens) and management with novel insecticide molecule Spinosad. The degree of resistance developed by resistant population of C. ferrugenius shows 34.66 and 21.35 folds resistant at LC50 level; 4.14 and 9.89 folds resistant at LC99.9 level to malathion and deltamethrin, respectively in comparison with laboratory susceptible strain at 72 HAT. Insecticide tested for the management of malathion and deltamethrin resistant population of C. ferrugenius, Spinosad was found to be the finest at LC99.9 level with the relative toxicity (3.43 and 1.48 times) than malathion and deltamethrin respectively.

First report of AChE1 (G119S) mutation and multiple resistance mechanisms in Anopheles gambiae s.s. in Nigeria

Susceptibility and PBO synergist bioassays were done using 3–5 days old female Anopheles mosquito collected from Lagos State, Nigeria with WHO test papers DDT (4%), permethrin (0.75%), Bendiocarb (1%) and PBO (4%) according to standard procedures. The activities of cytochrome P450s, glutathione S-transferase and carboxylesterases were determined using biochemical assays. The presence of kdr-w, kdr-e and Ace-1R mutations were examined using molecular assays. Resistance to DDT and permethrin in An gambiae s.s from the four Local Government Areas (LGAs) was recorded while suspected resistance to bendiocarb was recorded in mosquitoes from Alimosho and Kosofe LGAs. PBO synergist reduced the knockdown time and also recorded significantly (P < 0.05) higher 24 hrs percentage mortality compared to non-synergized bioassays. Increased activities of detoxifying enzymes was recorded in wild mosquito compared to the insecticides susceptible laboratory strain and this was significant (P < 0.05) in P450s, esterase α and β. Kdr-w was detected in An. gambiae s.s from all the LGAs, kdr-e (L1014S) was detected in Alimosho, Kosofe and Ibeju-Lekki, while the Ace-1R gene was detected in Alimosho and Kosofe. Results from this study provide evidence for resistance of An. gambiae from Lagos State to multiple classes of neurotoxic insecticides with multiple resistance mechanisms to these insecticides.