Piperonyl Butoxide Research Articles

Toxicity of isocycloseram, an isoxazoline insecticide, against laboratory and field-collected German cockroaches (Blattodea: Ectobiidae).

Isocycloseram is a new insecticide in the isoxazoline class that targets insect GABA-gated chloride channels. In this study, we evaluated a cockroach gel bait formulation containing 1% isocycloseram against a susceptible strain (UCR) and 5 field-collected strains (WM, RG386, Ryan, CDR, and SY) of the German cockroach, Blattella germanica (L.) (Blattodea: Ectobiidae), and compared it with several commercial insecticide baits in the laboratory. Using the Ebeling choice box method, we also tested a residual deposit of an SC formulation of isocycloseram against the UCR, RG386, and Ryan strains. The isocycloseram bait was among the fastest-performing treatments against adult males (mean survival time: 0.9-2.7 days) and mixed stages and sexes (mean survival time: 1.4-5.4 days) across all strains. Secondary transfer effects of the bait were demonstrated in the UCR strain by exposing new adult males to individuals killed by direct bait treatment. Physiological resistance was not detected in the WM, CDR, and RG386 strains with topical treatment of a diagnostic dose (3× LD95) of isocycloseram developed using the UCR strain. However, topical assays revealed resistance ratios (RR50) of 1.6 and 3.0× in the Ryan and SY strains, respectively. The performance of a 0.05% isocycloseram residual application against the Ryan strain was improved with the addition of piperonyl butoxide.

Rifampicin synergizes the toxicity of insecticides against the green peach aphid, Myzus persicae

Myzus persicae is an important pest that has developed resistance to nearly all currently used insecticidal products. The employment of insecticide synergists is one of the effective strategies that need to be developed for the management of this resistance. Our study showed that treatment with a combination of the antibiotic, rifampicin, with imidacloprid, cyantraniliprole, or clothianidin significantly increased their toxicities against M. persicae, by 2.72, 3.59, and 2.41 folds, respectively. Rifampicin treatment led to a noteworthy reduction in the activities of multifunctional oxidases (by 32.64%) and esterases (by 23.80%), along with a decrease in the expression of the CYP6CY3 gene (by 58.57%) in M. persicae. It also negatively impacted the fitness of the aphids, including weight, life span, number of offspring, and elongation of developmental duration. In addition, bioassays showed that the combination of rifampicin and a detoxification enzyme inhibitor, piperonyl butoxide, or dsRNA of CYP6CY3 further significantly improved the toxicity of imidacloprid against M. persicae, by 6.19- and 7.55-fold, respectively. The present study suggests that development of active ingredients such as rifampicin as candidate synergists, show promise to overcome metabolic resistance to insecticides in aphids.

The effectiveness of acaricidal drugs in bovine psoroptosis

The report presents the results of acaricides using from the macrocyclic lactones group and complex drug administration which includes deltamethrin, diflubenzuron and piperonyl butoxide by psoroptic scab of cattle. There was research at the beefproducing farm (Aberdeen Angus breed). It was four groups (n=10 by each). Animals of the 1st experimental group have received «Meradoc» drug (doramectin 1%, subcutaneous introduction, the neck area), 2d experimental group – «Ivermec» drug (ivermectin 1%, 1 ml per 50 kg, subcutaneous introduction, the neck area), 3d experimental group – «Deltsid 7.5» by single dose (15 ml per 100 kg) by the application on skin vertebral from the head to the tail. The 4th group was control without acaricidal treatment. It was found the single using of macrocyclic lactones groups’ drugs were destroyed only the larval, nymphal and imaginal phases of acarian Psoroptes without ticks’ eggs extermination. The scraping was included ticks’ eggs on 7th day after «Deltsid 7.5» drug using, but it wasn’t detected any larval, imaginal phases and ticks’ eggs on the 14th, 21st and 28th days after beginning of animal’s treatment. Moreover, to the 28th days there were registered the hair restoration and the skin in affected areas without compaction and folding. The using of complex drug «Deltsid 7.5» is more effective treatment of psoroptic scab of cattle. This drug has the acaricidal effect on the preimaginal phases Acariformes. There were blocked the enzymes activity which destroy the drug’s effect. To take on the Acariformes resistibility by acaricides it should be control the quality of anti-acaricidal treatments by microscopy of skin scrapings. The recommendation in the case of the positive result’s getting is the treatment alternation.

Application of insecticide dusts for the management of Indian meal moth (Lepidoptera: Pyralidae) wandering larvae under laboratory conditions.

The Indian meal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), is a very common stored-product pest. The mature larvae wander around searching for suitable places to pupate, offering an opportunity to control this pest. We evaluated the efficacy of 5 insecticide dusts: Alpine (0.25% dinotefuran, 95% diatomaceous earth [DE]), CimeXa (92.1% amorphous silica gel), DX13 (100% DE), Tempo (1% cyfluthrin), and Tri-Die (1% pyrethrin, 10% piperonyl butoxide, 40% amorphous silica) against P. interpunctella wandering larvae by allowing larvae to pass 1-inch treated band, expose to treated vinyl tiles for 5min, and expose to treated harborage (with or without the presence of an untreated harborage). A commercially treated harborage product (Nattaro band) served as a positive control in the exposure to the treated harborage test. Tempo was significantly more effective than other insecticide dusts in the 1-inch band and forced exposure tests but caused only 44% and 54% larvae mortality in the 2 tests. In contrast, CimeXa, Tri-Die, and Tempo caused 84%-89% mortality when the larvae were provided with treated harborages. When both treated and untreated harborages were present, Tempo caused a significantly higher mortality and a lower percentage of emerged adults from larvae than Tri-Die, but not significantly more than CimeXa. Deploying Tempo or CimeXa-treated harborages and/or applying insecticide dust directly into the wall crevices, perimeters of the floor, shipping pallets, and other areas where P. interpunctella larvae hide could be an effective method for the management of the wandering stage of P. interpunctella larvae in storage facilities.

Temperature, mosquito feeding status and mosquito density influence the measured bio-efficacy of insecticide-treated nets in cone assays

BackgroundThe WHO cone bioassay is routinely used to evaluate the bioefficacy of insecticide-treated nets (ITNs) for product pre-qualification and confirmation of continued ITN performance during operational monitoring. Despite its standardized nature, variability is often observed between tests. We investigated the influence of temperature in the testing environment, mosquito feeding status and mosquito density on cone bioassay results.MethodsCone bioassays were conducted on MAGNet (alphacypermethrin) and Veeralin (alphacypermethrin and piperonyl butoxide (PBO)) ITNs, using laboratory-reared pyrethroid-resistant Anopheles funestus sensu stricto (FUMOZ strain) mosquitoes. Three experiments were conducted using standard cone bioassays following WHO-recommended test parameters, with one variable changed in each bioassay: (i) environmental temperature during exposure: 22–23 °C, 26–27 °C, 29–30 °C and 32–33 °C; (ii) feeding regimen before exposure: sugar starved for 6 h, blood-fed or sugar-fed; and (iii) mosquito density per cone: 5, 10, 15 and 20 mosquitoes. For each test, 15 net samples per treatment arm were tested with four cones per sample (N = 60). Mortality after 24, 48 and 72 h post-exposure to ITNs was recorded.ResultsThere was a notable influence of temperature, feeding status and mosquito density on An. funestus mortality for both types of ITNs. Mortality at 24 h post-exposure was significantly higher at 32–33 °C than at 26–27 °C for both the MAGNet [19.33% vs 7%; odds ratio (OR): 3.96, 95% confidence interval (CI): 1.99–7.87, P < 0.001] and Veeralin (91% vs 47.33%; OR: 22.20, 95% CI: 11.45–43.05, P < 0.001) ITNs. Mosquito feeding status influenced the observed mortality. Relative to sugar-fed mosquitoes, The MAGNet ITNs induced higher mortality among blood-fed mosquitoes (7% vs 3%; OR: 2.23, 95% CI: 0.94–5.27, P = 0.068) and significantly higher mortality among starved mosquitoes (8% vs 3%, OR: 2.88, 95% CI: 1.25–6.63, P = 0.013); in comparison, the Veeralin ITNs showed significantly lower mortality among blood-fed mosquitoes (43% vs 57%; OR: 0.56, 95% CI: 0.38–0.81, P = 0.002) and no difference for starved mosquitoes (58% vs 57%; OR: 1.05, 95% CI: 0.72–1.51, P = 0.816). Mortality significantly increased with increasing mosquito density for both the MAGNet (e.g. 5 vs 10 mosquitoes: 7% vs 12%; OR: 1.81, 95% CI: 1.03–3.20, P = 0.040) and Veeralin (e.g. 5 vs 10 mosquitoes: 58% vs 71%; OR 2.06, 95% CI: 1.24–3.42, P = 0.005) ITNs.ConclusionsThe results of this study highlight that the testing parameters temperature, feeding status and mosquito density significantly influence the mortality measured in cone bioassays. Careful adherence to testing parameters outlined in WHO ITN testing guidelines will likely improve the repeatability of studies within and between product testing facilities.Graphical

EVALUATION OF THERMAL FOGGER FOR EFFECTIVENESS OF LARVICIDE-ADULTICIDE MIXTURE AGAINST AEDES AEGYPTI

A hand-held thermal fogger was evaluated to spray a mixture of adulticide (Aqualuer 20-20® containing permethrin and piperonyl butoxide) and a biological larvicide (VectoBac®12AS, containing Bacillus thuringiensis subspecies israelensis). Adulticide was applied at maximum label rate of 37 mL/ha while larvicide rates were 0.58, 1.76, 2.64, and 3.50 L/ha (Maximum label rate 2.34 L/ha). Two rows, 7.6 m apart, of bioassay cages containing susceptible 25, 5-7 days old female Aedes aegypti were used to assess effectiveness of the adulticide. Along with each cage were two 473 mL empty plastic cups at 0 and 1.3 m above ground to collect the larvicide. Two control cages and two empty cups were placed upwind in the field for five minutes and removed before the spray. The study was conducted in the evening with five separated replications of a rate. The exposed cages and cups were retrieved from the field and brought back to the laboratory for further observations and tests to determine mortality at 24 and 48 h, respectively. All the adults in all treatment cages and all tests died within 24 h of the spray application. In the lab, larval mortality was assessed by introducing Ae. aegypti larvae, where mortality decreased with increasing distance from the spray line and increased with increasing application rate. The larval mortality at all distances was strongly correlated with application rate (R2 = 0.98). The highest rate used produced 64 -95% larval mortality at all distances. The results indicate that Aqualuer 20-20 and VectoBac 12AS can be applied simultaneously as a mixture achieving complete adult control and considerable larval control.

Sublethal effect and detoxifying metabolism of metaflumizone and indoxacarb on the fall armyworm, Spodoptera frugiperda

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) (Lepidoptera, Noctuidae), is a highly polyphagous invasive pest that damages various crops. Pesticide control is the most common and effective strategy to control FAW. In this study, we evaluated the toxicity of metaflumizone and indoxacarb against third-instar FAW larvae using the insecticide-incorporated artificial diet method under laboratory conditions. Both metaflumizone and indoxacarb exhibited substantial toxicity against FAW, with LC50 values of 2.43 and 14.66 mg/L at 72 h, respectively. The sublethal effects of metaflumizone and indoxacarb on parental and F1 generation FAW were investigated by exposing third-instar larvae to LC10 and LC30 concentrations of these insecticides. Sublethal exposure to these two insecticides significantly shortened adult longevity, extended pupal developmental times and led to reduced pupal weight, pupation rates, and adult fecundity in the treated parental generation and F1 generation at LC10 or LC30 concentrations, in comparison to the control group. The larval developmental times were shortened in the parental generation but prolonged in the F1 generation, after being treated with sublethal concentrations of metaflumizone. Furthermore, larvae exposed to LC10 or LC30 concentrations of indoxacarb exhibited elevated activity levels of cytochrome P450 monooxygenase and glutathione S-transferase, which coincides with the observed synergistic effect of piperonyl butoxide and diethyl maleate. In conclusion, the high toxicity and negative impact of metaflumizone and indoxacarb on FAW provided significant implications for the rational utilization of insecticides against this pest.

Insecticide susceptibility status of Anopheles gambiae mosquitoes and the effect of pre-exposure to a piperonyl butoxide (PBO) synergist on resistance to deltamethrin in northern Namibia.

Pyrethroid-based indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs) have been employed as key vector control measures against malaria in Namibia. However, pyrethroid resistance in Anopheles mosquitoes may compromise the efficacy of these interventions. To address this challenge, the World Health Organization (WHO) recommends the use of piperonyl butoxide (PBO) LLINs in areas where pyrethroid resistance is confirmed to be mediated by mixed function oxidase (MFO). This study assessed the susceptibility of Anopheles gambiae sensu lato (s.l.) mosquitoes to WHO tube bioassays with 4% DDT and 0.05% deltamethrin insecticides. Additionally, the study explored the effect of piperonyl butoxide (PBO) synergist by sequentially exposing mosquitoes to deltamethrin (0.05%) alone, PBO (4%) + deltamethrin (0.05%), and PBO alone. The Anopheles mosquitoes were further identified morphologically and molecularly. The findings revealed that An. gambiae sensu stricto (s.s.) (62%) was more prevalent than Anopheles arabiensis (38%). The WHO tube bioassays confirmed resistance to deltamethrin 0.05% in the Oshikoto, Kunene, and Kavango West regions, with mortality rates of 79, 86, and 67%, respectively. In contrast, An. arabiensis displayed resistance to deltamethrin 0.05% in Oshikoto (82% mortality) and reduced susceptibility in Kavango West (96% mortality). Notably, there was reduced susceptibility to DDT 4% in both An. gambiae s.s. and An. arabiensis from the Kavango West region. Subsequently, a subsample from PBO synergist assays in 2020 demonstrated a high proportion of An. arabiensis in Oshana (84.4%) and Oshikoto (73.6%), and 0.42% of Anopheles quadriannulatus in Oshana. Non-amplifiers were also present (15.2% in Oshana; 26.4% in Oshikoto). Deltamethrin resistance with less than 95% mortality, was consistently observed in An. gambiae s.l. populations across all sites in both 2020 and 2021. Following pre-exposure to the PBO synergist, susceptibility to deltamethrin was fully restored with 100.0% mortality at all sites in 2020 and 2021. Pyrethroid resistance has been identified in An. gambiae s.s. and An. arabiensis in the Kavango West, Kunene, and Oshikoto regions, indicating potential challenges for pyrethroid-based IRS and LLINs. Consequently, the data highlights the promise of pyrethroid-PBO LLINs in addressing resistance issues in the region.

Efficacy of pesticides against Nesidiocoris tenuis Reuter (Hemiptera: Miridae), an emerging threat in the cultivation of tomato in Northwest Europe

As non-chemical methods are not always effective to manage pests, the search for chemical solutions that efficiently combat pest species, while safeguarding the environment and beneficial organisms, is essential for advancing effective Integrated Pest Management (IPM) programs. Here, we investigated the efficacy of commercially available pesticides in managing Nesidiocoris tenuis, an emerging threat in the cultivation of tomato in Northwest Europe, while preserving the biocontrol agent Macrolophus pygmaeus. Fourteen pesticides were assessed for their residual toxicity on adults and nymphs in laboratory experiments, after which the most promising candidate was evaluated in cages and semi-commercial greenhouse conditions. Among the pesticides tested, chlorantraniliprole, hexythiazox, tebufenozide and azadirachtin had minimal impact on both insect species, with less than 37.5% corrected mortality. Pirimicarb, sulfoxaflor, acetamiprid, flupyradifurone, thiacloprid, tebufenpyrad, spinosad, indoxacarb, and a mixture of pyrethrins and piperonyl butoxide were harmful to N. tenuis, but also affected M. pygmaeus (up to 100% mortality). Emamectin benzoate was moderately effective to N. tenuis adults (87%) and nymphs (80%), but had limited effects on M. pygmaeus. Application of emamectin benzoate at the maximum recommended concentration resulted in a significantly higher mortality of N. tenuis adults (85%) compared to M. pygmaeus (53%) in cage experiments, while it significantly reduced N. tenuis nymphs and the number of necrotic rings in a greenhouse experiment. Emamectin benzoate emerges as a promising candidate for inclusion in IPM programs. However, further research is necessary to determine its compatibility with other management strategies for controlling N. tenuis and other tomato pests.

Monitoring Aedes populations for arboviruses, Wolbachia, insecticide resistance and its mechanisms in various agroecosystems in Benin

Aedes mosquitoes are the main vectors of arboviruses in Benin. Cases of dengue have been reported in Benin with all four serotypes of the virus actively circulating in this region. Some agricultural settings are known to harbor Aedes vectors responsible for the transmission of arboviruses. The massive use of certain insecticides in agricultural settings has probably contributed to insecticide resistance in these vectors. In Benin, the susceptibility of arbovirus vectors to insecticides is poorly studied. In addition, the distribution of Wolbachia spp., which is used against some arboviruses is unknown. Moreover, there is limited information regarding the vectors responsible for the transmission of arboviruses in Benin. This present study monitored the species composition, arboviruses, and Wolbachia symbiont status, as well as the phenotypic and molecular insecticide resistance profile of Aedes populations from three agroecosystems in Benin. Aedes species identification was performed morphologically and confirmed using qPCR. (RT)-qPCR assay was applied for monitoring the presence of DENV, CHIKV, ZIKV, and WNV pathogens as well as for naturally occurring Wolbachia symbionts. Insecticide resistance was assessed phenotypically, by permethrin (0.75%) exposure of Adults (F0) using World Health Organization (WHO) bioassay protocols, and at the molecular level, using TaqMan (RT)-qPCR assays for assessing knock-down resistance (kdr) mutations (F1534C, V1016G/I, and S989P) and the expression levels of eight detoxification genes (P450s from the CYP9 and CYP6 families, carboxylesterases and glutathione-S-transferases). Aedes aegypti (Ae. aegypti) mosquitoes were the most abundant (93.9%) in the three agroecosystems studied, followed by Aedes albopictus (Ae. albopictus) mosquitoes (6.1%). No arboviruses were detected in the study's mosquito populations. Naturally occurring Wolbachia symbionts were present in 7 pools out of 15 pools tested. This could influence the effectiveness of vector control strategies based on exogenously introduced Wolbachia, all present in the three agroecosystems. Full susceptibility to permethrin was observed in all tested populations of Ae. albopictus. On the contrary, Ae. aegypti were found to be resistant in all three agroecosystem sites except for banana plantation sites, where full susceptibility was observed. Molecular analysis revealed that individual target site resistance kdr mutations F1534C and V1016G/I were detected in most Ae. aegypti populations. Additionally, double mutant (F1534C + V1016G/I) mosquitoes were found in some populations, and in one case, triple mutant (F1534C + V1016G/I + S989P) mosquitoes were detected. Metabolic resistance, as reflected by overexpression of three P450 genes (CYP6BB2, CYP9J26, and CYP9J32), was also detected in Ae. aegypti mosquitoes. Our study provides information that could be used to strategize future vector control strategies and highlights the importance of continuing vector surveillance. Future studies should assess the effect of piperonyl butoxide (PBO) on metabolic resistance and identify the different strains of Wolbachia spp., to choose the best vector control strategies in Benin.

Time of exposure and assessment influence the mortality induced by insecticides against metabolic resistant mosquitoes

BackgroundIncreasing metabolic resistance in malaria vector mosquitoes resulted in the development of insecticide-treated nets (ITNs) with active ingredients (AI) that target them. Bioassays that accurately measure the mortality induced by these AIs on ITNs are needed. Mosquito metabolic enzyme expression follows a circadian rhythm. Thus, this study assessed (i) influence of the time of day of mosquito exposure and (ii) timing of assessment of mortality post exposure (24 and 72 h) to ITNs against vectors that are susceptible to pyrethroids and those with metabolic and knockdown resistance mechanisms.MethodsTwo cone bioassay experiments were conducted following World Health Organization (WHO) guidelines. Firstly, on ITNs incorporated with 2 g AI/kg of deltamethrin (DM) alone, or combined with 8 g AI/kg piperonyl butoxide (PBO) synergist, during the day (9:00–14:00 h) and repeated in the evening (18:00–20:00 h). This was followed by a confirmatory experiment during the afternoon (12:00–14:00 h) and repeated in the night (22:00–24:00 h) using mosquitoes unexposed or pre-exposed to PBO for 1 h before exposure to DM ITNs. Each net piece was tested with a minimum of eight cones per time (N = 24). The outcome was mortality after 24 h (M24) or 72 h (M72) of holding.ResultsThe cone bioassays performed using metabolic resistant mosquitoes during the evening showed significantly lower M24 than those performed in the day for DM: odds ratio (OR) 0.14 [95% confidence interval (CI) 0.06–0.30, p < 0.0001] and DM PBO [OR 0.29 (95% CI 0.18—0.49, p < 0.0001). M72 was higher than M24 for metabolic resistant mosquitoes exposed to DM [OR 1.44 (95% CI 1.09–1.88), p = 0.009] and DM PBO [OR 1.82 (95% CI 1.42–2.34), p < 0.0001]. An influence of hour of experiment and time of assessment was not observed for mosquitoes that had knockdown resistance or that were pyrethroid-susceptible.ConclusionsTime of day of experiment and hour of assessment of delayed mortality after exposure of mosquitoes are important considerations in evaluating insecticides that interact with mosquito metabolism to counter metabolic resistant mosquitoes. This is important when evaluating field-aged ITNs that may have lower concentrations of AI.Graphical

Adult mosquitoes of the sibling species Anopheles gambiae and Anopheles coluzzii exhibit contrasting patterns of susceptibility to four neonicotinoid insecticides along an urban-to-rural gradient in Yaoundé, Cameroon

BackgroundNeonicotinoids are potential alternatives for controlling pyrethroid-resistant mosquitoes, but their efficacy against malaria vector populations of sub-Saharan Africa has yet to be investigated. The aim of the present study was to test the efficacy of four neonicotinoids against adult populations of the sibling species Anopheles gambiae and Anopheles coluzzii sampled along an urban-to-rural gradient.MethodsThe lethal toxicity of three active ingredients for adults of two susceptible Anopheles strains was assessed using concentration–response assays, and their discriminating concentrations were calculated. The discriminating concentrations were then used to test the susceptibility of An. gambiae and An. coluzzii mosquitoes collected from urban, suburban and rural areas of Yaoundé, Cameroon, to acetamiprid, imidacloprid, clothianidin and thiamethoxam.ResultsLethal concentrations of neonicotinoids were relatively high suggesting that this class of insecticides has low toxicity against Anopheles mosquitoes. Reduced susceptibility to the four neonicotinoids tested was detected in An. gambiae populations collected from rural and suburban areas. By contrast, adults of An. coluzzii that occurred in urbanized settings were susceptible to neonicotinoids except acetamiprid for which 80% mortality was obtained within 72 h of insecticide exposure. The cytochrome inhibitor, piperonyl butoxide (PBO), significantly enhanced the activity of clothianidin and acetamiprid against An. gambiae mosquitoes.ConclusionsThese findings corroborate susceptibility profiles observed in larvae and highlight a significant variation in tolerance to neonicotinoids between An. gambiae and An. coluzzii populations from Yaoundé. Further studies are needed to disentangle the role of exposure to agricultural pesticides and of cross-resistance mechanisms in the development of neonicotinoid resistance in some Anopheles species.

(Non)targeted Chemical Analysis and Risk Assessment of Organic Contaminants in Darkibor Kale Grown at Rural and Urban Farms.

This study investigated the presence and human hazards associated with pesticides and other anthropogenic chemicals identified in kale grown in urban and rural environments. Pesticides and related compounds (i.e., surfactants and metabolites) in kale samples were evaluated using a nontargeted data acquisition for targeted analysis method which utilized a pesticide mixture containing >1,000 compounds for suspect screening and quantification. We modeled population-level exposures and assessed noncancer hazards to DEET, piperonyl butoxide, prometon, secbumeton, terbumeton, and spinosyn A using nationally representative estimates of kale consumption across life stages in the US. Our findings indicate even sensitive populations (e.g., pregnant women and children) are not likely to experience hazards from these select compounds were they to consume kale from this study. However, a strictly nontargeted chemical analytical approach identified a total of 1,822 features across all samples, and principal component analysis revealed that the kale chemical composition may have been impacted by agricultural growing practices and environmental factors. Confidence level 2 compounds that were ≥5 times more abundant in the urban samples than in rural samples (p < 0.05) included chemicals categorized as "flavoring and nutrients" and "surfactants" in the EPA's Chemicals and Products Database. Using the US-EPA's Cheminformatics Hazard Module, we identified that many of the nontarget compounds have predicted toxicity scores of "very high" for several end points related to human health. These aspects would have been overlooked using traditional targeted analysis methods, although more information is needed to ascertain whether the compounds identified through nontargeted analysis are of environmental or human health concern. As such, our approach enabled the identification of potentially hazardous compounds that, based on their hazard assessment score, merit follow-up investigations.

Antimicrobial Action of Sambacus nigra, Symphytum officinale, Urtica dioica, and Mentha pulegium Plant Compounds against Antibiotic Resistant Streptococcus pneumoniae

Infections by Streptococcus pneumoniae are among the leading causes of illness and death in children below 5 years, people with underlying debilitating medical conditions, and the elderly worldwide. Over the past three decades, antimicrobial resistance in S. pneumoniae has drastically increased, with resistance reported in Penicillin, erythromycin, trimethoprim/sulfamethoxazole, vancomycin, tetracycline, chloramphenicol and ofloxacin. This phenomenon has necessitated a continuous search for alternative compounds against S. pneumoniae, such as Plants. The objective was to investigate antimicrobial natural compounds present in S. nigra, S. officinale, U. dioica, and M. pulegium against antibiotic-resistant S. pneumoniae. The study utilised cross-sectional design where S. nigra leaves, S. officinale roots, U. dioica leaves, and M. pulegium leaves were collected from Kisii highland forests. Phytochemicals were identified by gas chromatography (GC/MS). Microbiological testing of the resistant strain of S. pneumoniae was done at Jaramogi Oginga Odinga Teaching and Referral Hospital Laboratory (JOOTRHL). The MBC and MIC were determined using disk diffusion. Data was analysed using ANOVA (P&lt;0.05). Results showed alkaloids, phenols, tannins, flavonoids, saponins, glycosides, steroids and terpenoids at different extraction solvent concentrations. S. nigra, U. dioica, S. officinale and M. pulegium phytochemical compounds showed significant differences at 0.825 &lt; 3.01. S. nigra and U. dioica showed considerable activity on resistant strains of S. pneumoniae. The MIC of S. nigra and U. dioica were 0.625 mg/L and 1.25 mg/L, while MBC was 0.3125 mg and 0.625 mg respectively. GC/MS analysis of phytochemical compounds on S. nigra and U. dioica showed a total of 8 NIST drug compounds. GC/MS study data; thymol, metharbital, Diethyltoluamide, Cyclohexanol, 5-methyl-2-(1-methylethyl)-, (1. alpha., 2. beta., 5. alpha.)-, aspirin, Piperonyl butoxide, triamterene and Glycine, N-benzoyl- are the most effective phytochemical compound in S. nigra and U. dioica against S. pneumoniae. The study concluded that S. nigra and U. dioica leaves pose active compounds that could be further used as alternatives in the treatment of antibiotic-resistant S. pneumoniae

Heavy Metals and Pesticide Residues in Small Farm Cheese Production in Croatia—Challenge between Quality and Quantity

The beneficial health effects of cheese as a source of bioactive compounds with antioxidant, antimicrobial, anti-inflammatory, immunomodulatory, and analgesic effects are well known. The aim of this study is to determine the presence of pesticide residues and heavy metals in 79 cheese samples from small farms in Croatia. The samples were analyzed over a period of three years for the quality parameters of fat, protein, dry matter, salt, and pH to determine whether metrological conditions affect the quality of cheese and to test the correlation between the variables in different types of cheese. A total of 509 pesticide residues were analyzed using liquid and gas chromatography with tandem mass spectrometry. Piperonyl butoxide was found in two samples. Inductively coupled plasma with mass spectrometry was used for a metal content analysis, and Cd, Cr, Mn, Ni, and Pb were found in the range of &lt;0.005–0.012 mg kg−1, &lt;0.02–0.84 mg kg−1, 0.031–1.128 mg kg−1, &lt;0.03–0.67 mg kg−1, and &lt;0.01–0.12 mg kg−1, respectively. Cd was detected in just three samples. Mn was found in all analyzed samples. All tested samples complied with EU regulations and directives, and at the point of analysis, none posed a direct health risk for consumers. Sustainability on small farms could be ensured with the responsible use of pesticides and through a consistent and reliable supply of fresh, high-quality milk.

Monitoring of Fabric Integrity and Attrition Rate of Dual-Active Ingredient Long-Lasting Insecticidal Nets in Tanzania: A Prospective Cohort Study Nested in a Cluster Randomized Controlled Trial.

Pyrethroid-treated long-lasting insecticidal nets (LLINs) have been the main contributor to the reduction in malaria in the past two decades in sub-Saharan Africa. The development of pyrethroid insecticide resistance threatens the future of LLINs, especially when nets become holed and pyrethroid decays. In this study, three new classes of dual-active ingredient (AI) LLINs were evaluated for their physical durability: (1) Royal Guard, combining pyriproxyfen, which disrupts female fertility, and a pyrethroid, alpha-cypermethrin; (2) Interceptor G2, which combines the pyrrole chlorfenapyr and a pyrethroid (alpha-cypermethrin); (3) Olyset Plus, which incorporates the pyrethroid permethrin and the synergist piperonyl butoxide, to enhance the pyrethroid potency; and Interceptor, a reference net that contains alpha-cypermethrin as the sole active ingredient. About 40,000 nets of each type were distributed in February 2019 to different villages in Misungwi. A total of 3072 LLINs were followed up every 6-12 months up to 36 months to assess survivorship and fabric integrity. The median functional survival was less than three years with Interceptor, Interceptor G2, and Royal Guard showing 1.9 years each and Olyset Plus showing 0.9 years. After 36 months, 90% of Olyset Plus and Royal Guard and 87% of Interceptor G2 were no longer in use (discarded) due to wear and tear, compared to 79% for Interceptor. All dual-AI LLINs exhibited poor textile durability, with Olyset Plus being the worst.

Resistance of mosquitoes to Lambda-Cyhalothrin and DDT in a Niger Delta Region of Nigeria.

The escalation of insecticide resistance across the World Health Organization (WHO) African region calls for the routine monitoring of insecticide resistance in mosquitoes. While pyrethroids are recommended for use and dichlorodiphenyltrichloroethane (DDT) has been prescribed, it is imperative to reevaluate their efficacy across diverse geographical settings. The extent of resistance among mosquitoes to DDT and lambda-cyhalothrin remains understudied in Ika North East, Delta State, Nigeria, where the yellow fever virus was reported in 2019. We investigated the susceptibility of female mosquitoes to DDT (4%) and lambda-cyhalothrin (0.05%) in this specific study area. Adult female mosquitoes of Anopheles gambiae, Culex quinquefasciatus, and Aedes albopictus, aged between 2 and 4 days, were subjected to these insecticides utilizing the WHO bioassay method. The assessment of knockdown was done between 10 and 60 min, then mortality after 24 h. C. quinquefasciatus and A. albopictus mosquitoes exposed to DDT and lambda-cyhalothrin exhibited high susceptibility, resulting in complete mortality (100%); however, A. gambiae displayed resistance, with mortality rates of 19% and 76%, respectively . Notably, A. gambiae mosquitoes exposed to piperonyl butoxide (PBO)-DDT and PBO-lambda-cyhalothrin exhibited enhanced mortality, reaching 95% (indicating suspected resistance) and 100% (indicating susceptibility), respectively. Knockdown time (KDT) for 50% in mosquitoes exposed to lambda-cyhalothrin ranged from 3.94 to 33.51 min. Similarly, KDT model for 95% ranged from 19.04 to 84.15 min. Among the tested mosquito species, Culex mosquitoes exhibited the shortest knockdown resistance time for lambda-cyhalothrin, recorded at 3.94 min. Similarly, the KDT for DDT ranged from 24.97 to 187.06 min for 50% mortality and from 61.04 to 431.03 min for 95% mortality, respectively. Anopheles mosquitoes exposed to PBO + DDT recorded the lowest KDT. Our study underscores the potential of exercising caution in the use of DDT and lambda-cyhalothrin insecticides for the control of mosquitoes due to emerging resistance.

Exploring the molecular mechanisms of increased intensity of pyrethroid resistance in Central African population of a major malaria vector Anopheles coluzzii.

Molecular mechanisms driving the escalation of pyrethroid resistance in the major malaria mosquitoes of Central Africa remain largely uncharacterized, hindering effective management strategies. Here, resistance intensity and the molecular mechanisms driving it were investigated in a population of Anopheles coluzzii from northern Cameroon. High levels of pyrethroid and organochloride resistance were observed in An. coluzzii population, with no mortality for 1× permethrin; only 11% and 33% mortalities for 5× and 10× permethrin diagnostic concentrations, and <2% mortalities for deltamethrin and DDT, respectively. Moderate bendiocarb resistance (88% mortality) and full susceptibility to malathion were observed. Synergist bioassays with piperonyl butoxide recovered permethrin susceptibility, with mortalities increasing to 53.39%, and 87.30% for 5× and 10× permethrin, respectively, implicating P450 monooxygenases. Synergist bioassays with diethyl maleate (DEM) recovered permethrin and DDT susceptibilities (mortalities increasing to 34.75% and 14.88%, respectively), implicating glutathione S-transferases. RNA-seq-based genome-wide transcriptional analyses supported by quantitative PCR identified glutathione S-transferase, GSTe2 (RNA-seqFC = 2.93 and qRT-PCRFC = 8.4, p < 0.0043) and CYP450, CYP6Z2 (RNA-seqFC = 2.39 and qRT-PCRFC = 11.7, p < 0.0177) as the most overexpressed detoxification genes in the pyrethroid-resistant mosquitoes, compared to mosquitoes of the susceptible Ngousso colony. Other overexpressed genes include P450s, CYP6M2 (FC = 1.68, p < 0.0114), CYP4G16 (FC = 2.02, p < 0.0005), and CYP4G17 (FC = 1.86, p < 0.0276). While high frequency of the 1014F kdr mutation (50%) and low frequencies of 1014S (6.61%) and 1575Y (10.29%) were observed, no ace-1 mutation was detected in bendiocarb-resistant populations, suggesting the preeminent role of metabolic mechanism. Overexpression of metabolic resistance genes (including GSTe2 and CYP6Z2 known to confer resistance to multiple insecticides) in An. coluzzii from the Sudan Savannah of Cameroon highlights the need for alternative management strategies to reduce malaria burden in northern Cameroon.

Combination Insecticide Treatments with Methoprene and Pyrethrin for Control of Khapra Beetle Larvae on Different Commodities.

Trogoderma granarium Everts, the khapra beetle, is a serious pest of stored products throughout the world. Larvae pose a significant threat to stored products because they feed on >100 different commodities, possess the ability to enter facultative diapause, and are difficult to detect. Control methods for T. granarium include fumigation, contact insecticides, trapping, and insecticide-incorporated packaging. The objective of this study was to determine the residual efficacy of two insecticide formulations (methoprene + deltamethrin + piperonyl butoxide synergist Gravista® and methoprene + deltamethrin, DiaconIGR®Plus). These insecticides were evaluated on three stored product commodities, corn, wheat, and brown rice, by exposing T. granarium larvae during a 12-month testing period. Both formulations significantly reduced adult emergence on corn and wheat for 12 months and on brown rice for up to 6 months. Adult emergence was highest at month 12 for corn (8.41%), and brown rice (85.88%), and month 9 for wheat (39.52%), treated with DiaconIGR®Plus or Gravista®, respectively. A biological index used to measure the development of exposed larvae on the treated grain from the larval stage (low values) to adult emergence (high values) was lower (fewer adults) on corn and wheat compared to controls. Despite differences in formulations, each of these grain protectants could be utilized by stored commodity managers to protect commodities during storage and transportation for T. granarium when and if this pest is detected at USA ports of entry.

Knock-in mutagenesis in Drosophila Rdl underscores the critical role of the conserved M3 glycine in mediating the actions of broflanilide and isocycloseram on GABA receptors

γ-Aminobutyric acid receptors (GABARs) are crucial targets for pest control chemicals, including meta-diamide and isoxazoline insecticides, which act as negative allosteric modulators of insect GABARs. Previous cell-based assays have indicated that amino acid residues in the transmembrane cavity between adjacent subunits of Drosophila RDL GABAR (i.e., Ile276, Leu280, and Gly335) are involved in mediating the action of meta-diamides. In this study, to confirm this result at the organismal level, we employed CRISPR/Cas9-mediated genome editing, generated six transgenic Drosophila strains carrying substitutions in these amino acid residues, and investigated their sensitivity to broflanilide and isocycloseram. Flies homozygous for the I276F mutation did not exhibit any change in sensitivity to the tested insecticides compared to the control flies. Conversely, I276C homozygosity was lethal, and heterozygous flies exhibited ∼2-fold lower sensitivity to broflanilide than the control flies. Flies homozygous for the L280C mutation survived into adulthood but exhibited infertility. Both heterozygous and homozygous L280C flies exhibited ∼3- and ∼20-fold lower sensitivities to broflanilide and isocycloseram, respectively, than the control flies. The reduction in sensitivity to isocycloseram in L280C flies diminished to ∼3-fold when treated with piperonyl butoxide. Flies homozygous for the G335A mutation reached the adult stage. However, they were sterile, had small bodies, and exhibited reduced locomotion, indicating the critical role of Gly335 in RDL function. These flies exhibited markedly increased tolerance to topically applied broflanilide and isocycloseram, demonstrating that the conserved Gly335 is the target of the insecticidal actions of broflanilide and isocycloseram. Considering the significant fitness costs, the Gly335 mutation may not pose a serious risk for the development of resistance in field populations of insect pests. However, more careful studies using insect pests are needed to investigate whether our perspective applies to resistance development under field conditions.