Resistance Risk Assessment Research Articles

Assessing the potential risk of human pathogen resistance to medical antifungal treatments arising from agricultural use of fungicides with the same mode of action

AbstractA mechanistic basis is described for assessment of resistance risk to medical antifungal treatments from agricultural use of fungicides of the same mode of action. The following need to occur in landscape environments for a risk to be posed by dual use: (i) emergence, whereby a resistant strain emerges by mutation and invasion; (ii) selection, whereby a mutation conferring a fitness advantage is selected for in the presence of fungicide; and (iii) exposure of humans to resistant strains from the landscape, potentially resulting in invasive fungal infection. We identify 20 human pathogens for which there is evidence that all three processes above could, in principle, occur. A model is derived to explore what determines resistance emergence and selection in human pathogens in landscape environments. Knowledge gaps are identified in key parameters. The analysis suggests that emergence and selection were particularly affected by fitness cost associated with the resistance mutation(s) and fungicide concentration. Emergence was also determined by the amount of pathogen reproduction (related to pathogen population size). If fungicide resistance is associated with even a small fitness cost, then environments with low fungicide concentrations, such as field soils, may not be conducive to resistance emergence or selection. These general findings were related to a specific case of observational data from the Netherlands for Aspergillus fumigatus. The analysis supports previous work that compost is towards the high‐risk end of the spectrum for this species. Agricultural soils, nonagricultural land and grassland were lower risk.

Risk assessment of broflanilide resistance in Panonychus citri (McGregor): Cross-resistance, inheritance and relative fitness

Risk assessment of broflanilide resistance in Panonychus citri (McGregor): Cross-resistance, inheritance and relative fitness

Resistance risk assessment and fitness cost of cyproflanilide in the common cutworm, Spodoptera litura (Lepidoptera: Noctuidae)

As a major crop pest, common cutworm (CCW) Spodoptera litura (Fabricius) has evolved resistance to several insecticides. Cyproflanilide is a novel meta-diamide insecticide targeting the γ-aminobutyric acid receptor (GABAR), and displays excellent insecticidal activity against lepidopteran pests, including CCW. Therefore, the resistance risk of CCW against cyproflanilide was assessed and the fitness cost was evaluated as well. After exposure with cyproflanilide for 10 generations (F10), the LC50 values of CCW against cyproflanilide increased 1.26 - fold. When the realized heritability (h2) to cyproflanilide in the field was assumed to be the laboratory-estimated value (h2 = 0.09) and the mortality of CCW was 50–90 %, 126–57 generations were required to obtain a 10-fold resistance against cyproflanilide. After selected with cyproflanilide for 10 generations, the egg duration, larval duration, pre-pupal duration and female pupal weight were significantly increased, but the pupal duration, pupation rate and life-cycle rate were significantly decreased. The mean generation time (T) of F10 was significantly prolonged by 4.39 days. The value of relative fitness (Rf) in F10 was 0.99. Although resistance developed slowly, the prolonged egg and larval durations have implications for managing CCW in the field. This information will contribute to identifying practices to delay the resistance development of CCW to cyproflanilide.

Research on Resistance Monitoring, Risk Assessment, and Mechanisms of <i>Megalurothrips usitatus</i>

<i>Megalurothrips usitatus</i>, also known as common thrips or cowpea thrips, is a widely distributed and highly destructive pest, primarily infesting legume crops. Due to its short generation cycle, high reproductive capacity, and concealed lifestyle, the effectiveness of chemical pesticide control has been continuously diminishing with the modernization of agricultural production and the extensive use of pesticides. <i>Megalurothrips usitatus</i> has gradually developed resistance to various commonly used pesticides, with resistance levels increasing year by year, thus exacerbating the difficulty of pest management and causing significant economic losses to agricultural production. Scholars, both domestically and internationally, have conducted in-depth research using methods from morphology, molecular biology, and ecology, showing that there are three main causes of pest resistance: enhanced detoxification enzymes, reduced sensitivity at target sites, and decreased cuticle penetration. These findings provide a wealth of theoretical support for resistance monitoring and management. The resistance of <i>Megalurothrips usitatus</i> to multiple pesticides is not only a local issue but also affects global agricultural sustainability. Research on the monitoring, risk assessment, and mechanisms of resistance in <i>Megalurothrips usitatus</i> contributes to prolonging the effective use of pesticides, improving control outcomes, and enhancing both the yield and quality of cowpea crops. These studies also provide a scientific basis for developing more effective control strategies and ensuring sustainable agricultural development.

Rapid identification of antibiotic resistance gene hosts by prescreening ARG-like reads

Effective risk assessment and control of environmental antibiotic resistance depend on comprehensive information about antibiotic resistance genes (ARGs) and their microbial hosts. Advances in sequencing technologies and bioinformatics have enabled the identification of ARG hosts using metagenome-assembled contigs and genomes. However, these approaches often suffer from information loss and require extensive computational resources. Here we introduce a bioinformatic strategy that identifies ARG hosts by prescreening ARG-like reads (ALRs) directly from total metagenomic datasets. This ALR-based method offers several advantages: (1) it enables the detection of low-abundance ARG hosts with higher accuracy in complex environments; (2) it establishes a direct relationship between the abundance of ARGs and their hosts; and (3) it reduces computation time by approximately 44–96% compared to strategies relying on assembled contigs and genomes. We applied our ALR-based strategy alongside two traditional methods to investigate a typical human-impacted environment. The results were consistent across all methods, revealing that ARGs are predominantly carried by Gammaproteobacteria and Bacilli, and their distribution patterns may indicate the impact of wastewater discharge on coastal resistome. Our strategy provides rapid and accurate identification of antibiotic-resistant bacteria, offering valuable insights for the high-throughput surveillance of environmental antibiotic resistance. This study further expands our knowledge of ARG-related risk management in future.

Resistance risk assessment of Rhizoctonia solani to four fungicides.

Hexaconazole, thifluzamide, difenoconazole and azoxystrobin are widely used fungicides for the control of Rhizoctonia solani in China. However, few studies have assessed the sensitivity and resistance risk of R. solani to these four fungicides. The sensitivities of 126 R. solani isolates to hexaconazole, thifluzamide, difenoconazole and azoxystrobin were determined, with average half maximal effective concentration (EC50) values of 0.0386, 0.0659, 0.663 and 1.508 μg mL-1, respectively. Field resistance monitoring of the four fungicides showed that the three isolates had moderate resistance to difenoconazole. Resistant mutants to the four fungicides were obtained by fungicide adaptation, and resistance could be stably inherited by most mutants. Compared with those of the parent isolates, the biological characteristics of hexaconazole-resistant mutants exhibited enhanced or similar compound fitness index (CFI), whereas most of the other mutants displayed reduced or comparable CFI. There was evidence of positive cross-resistance between hexaconazole and difenoconazole. In the presence of fungicides, the expression of the CYP51 genes in hexaconazole- and difenoconazole-resistant mutants significantly increased, the expression of SDH genes in thifluzamide-resistant mutants significantly decreased, and the expression of the Cyt b gene in azoxystrobin-resistant mutants did not significantly change. Based on these data, we speculated that R. solani had a low-to-medium resistance risk to four fungicides. The change of target gene expression may be one of the reasons for fungicide resistance in R. solani. This study provides a theoretical basis for monitoring resistance emergence and developing resistance management strategies to control R. solani. © 2024 Society of Chemical Industry.

The variation of resistome, mobilome and pathogen in domestic and industrial wastewater treatment systems

Wastewater treatment plants (WWTPs), including both domestic and industrial facilities, are key contributors to antibiotic resistance genes (ARGs) and human pathogens in the environment. However, the characteristics and dissemination mechanisms of ARGs in domestic (SD) and industrial (SI) wastewater treatment systems remain unclear, leading to uncertainties in risk assessment. Based on metagenomic analysis, we observed significant differences in the compositions of resistome (ARGs and metal resistance genes, MRGs), mobilome (mobile genetic elements, MGEs), and bacterial community between SD and SI. SI exhibited lower diversity of ARGs but higher abundance of MRGs compared to SD. The removal efficiency of resistome was lower in the SI than that in the SD. MGEs emerged as the primary driver of ARG dissemination in the WWTPs, followed by the bacterial community. Environmental conditions (physicochemical parameters, heavy metals, and antibiotics) indirectly influenced the variation of resistome. Significantly, environmental conditions and MGEs highly influenced the composition of resistome in the SI, while bacterial community more associated with resistome in the SD. Additionally, we identified 36 human bacterial pathogens as potential hosts of ARGs, MRGs, and MGEs in wastewater samples. This study provides new insights on the dissemination mechanisms and risk assessment of antimicrobial resistance in the different types of WWTPs.

Quantitative release assessment for beta-lactamase-producing Escherichia coli of dairy origin into vegetables

Abstract Outbreaks of food poisoning associated with vegetables contaminated with Escherichia coli have been reported globally. This study was conducted to assess the probability of releasing beta-lactamase-producing E . coli of dairy farm origin into vegetables in Japan. A release assessment model connecting dairy farms to vegetable farms was developed. Data on the indicated diseases and antimicrobial use in dairy cattle were obtained from the agricultural insurance program in Hokkaido Prefecture between 2016 and 2019. Data on bla -harboring E . coli in cattle were obtained from the Japan Veterinary Antimicrobial Resistance Monitoring System. Microbiological field sampling was conducted at dairy and vegetable farms. Inoculation experiments in vegetables were conducted. All processes in the model simulation were iterated for 1000 times. The estimated proportion of dairy farms holding cattle with bla -harboring E . coli was 6.00% (95% CI: 4.81–7.35%). Beta-lactams were used in 71.76% (333,098/464,204 average annual cases) of indicated diseases. The estimated concentration of bla -harboring E . coli in mixed fresh manure at affected farms was 3.33 log 10 CFU/g (95% CI: 3.33–3.34). The concentrations were reduced to 0.83 (95% CI: 0.34–1.25) and −0.54 log 10 CFU/g (95% CI: −1.04 to −0.05) in soil after immature manure, and slurry spraying, respectively. The estimated concentrations in the soil of fields of radish, leafy vegetables, tomato, and spring-seeded and autumn-seeded onions at harvest were −1.99 (95% CI: −2.48 to −1.57), −2.87 (95% CI: −3.36 to −2.42), −3.82 (95% CI: −4.32 to −3.40), −5.36 (95% CI: −5.85 to −4.93), and −5.94 (95% CI: −6.43 to −5.52) log 10 CFU/g, respectively. The concentrations in the bodies of leafy green spinach and lettuce were −9.88 (95% CI: −10.38 to −9.43) and −10.91 (95% CI, −11.52 to −8.05) log 10 CFU/g, respectively. The probability of ingesting bla -harboring E . coli of dairy origin with raw vegetables in Japan was thus assessed to be very low. One Health impact statement This study applied a release assessment in the World Organisation for Animal Health framework for antimicrobial resistance risk assessment to connect the antimicrobial use for dairy cattle, manure production and vegetable farming. As vegetable is often consumed raw, the assessment is directly associated with human health. Although the environmental continuum was not explored, the reduction of the hazard on the soil of vegetable farms was considered. The study involved the engagement with dairy and vegetable farmers and the agriculture industry in understanding farming behavior and value chain. The approach in this study can be applied to the complex problems across the animal-human-environment interface.

Risk assessment and transmission of fluoroquinolone resistance in drug-resistant pulmonary tuberculosis: a retrospective genomic epidemiology study

Fluoroquinolone resistance is a major challenge in treating Multidrug-Resistant Tuberculosis globally. The GenoType MTBDRsl Ver 2.0, endorsed by the WHO, was used to characterize fluoroquinolone resistance. The fluoroquinolone resistance rates in the MDR-TB, Rifampicin-Resistant TB, and non-MDR-TB were 33%, 16.5%, and 5.4%, respectively. The most common mutation found in fluoroquinolone-resistant isolates was D94G (49.5%) in the gyrA gene. Of the 150 MDR-TB isolates, the prevalence of Extensively Drug-Resistant Tuberculosis and pre-XDR-TB was 1.33% and 30%, respectively. Among the 139 RR-TB isolates, pre-XDR-TB prevalence was 15.8%. The fluoroquinolone resistance rates were 5.12% among the 1230 isoniazid-monoresistant isolates. The study found that MDR-TB and RR-TB have higher risk of fluoroquinolone resistance than non-MDR tuberculosis. Rifampicin-resistant isolates with a mutation at codon S450L have a higher risk (RR = 12.96; 95%CI: 8.34–20.13) of developing fluoroquinolone resistance than isolates with mutations at other codons in the rpoB gene. Isoniazid-resistant isolates with a mutation at codon S315T have a higher risk (RR = 2.09; 95%CI: 1.25–3.50) of developing fluoroquinolone resistance. The study concludes that rapid diagnosis of fluoroquinolone resistance before starting treatment is urgently needed to prevent the spread and increase of resistance and to achieve better treatment outcomes in areas where it is higher.

Antibiotic resistance risk assessment in Mymensingh, Bangladesh: Current scenario from human-animal-environmental interfaces viewpoint

The imprudent use of antibiotics increases the environmental microflora's resistance to various drugs, particularly antibiotics. Prescription data is crucial for understanding antibiotic usage frequency and dosage. This health-focused study aims to analyze antibiotic prescription patterns in human and veterinary practices to identify emerging trends in environmental antibiotic resistance.For this survey, A total of 6550 prescriptions were randomly collected from hospitals and pharmacies in Mymensingh sadar upazila, Bangladesh, between August and October 2022. Of these, 5123 (78 %) were for human cases and 1427 (22 %) for veterinary purposes. Photos of the prescriptions were taken and analyzed to understand prescribing habits. Additionally, 30 water samples from rivers, ponds, sewage, and households in Mymensingh City Corporation were collected to assess environmental antibiotic levels and resistance patterns of microorganisms.The analysis showed that Cephalosporins were the most prescribed antibiotics, found in 570 (56.27 %) of human prescriptions and 230 (42.99 %) of veterinary prescriptions. Aminoglycosides had the lowest frequency, with 13 (1.2 %) for humans and 46 (8.6 %) for animals. Macrolides (12.24 %), carboxylic acids (1.87 %), and rifamycins (1.28 %) were only found in human prescriptions, while sulfa drugs (10.84 %), tetracyclines (5.42 %), and combinations of antibiotics (14.77 %) were only in animal prescriptions. Quinolones were prescribed 4.06 times more for humans, while aminoglycosides were used 3.54 times more for animals. Environmental samples showed E. coli had the highest resistance (MAR Value: 0.625) against eight antibiotics.This study illuminates the human-animal prescription patterns that are influenced by environmental factors which drive antibiotic stewardship in Bangladesh. It is imperative for practitioners to exercise caution and adhere to guidelines when prescribing antibiotics, both in human and veterinary practices, given the alarming trend of antibiotic resistance. Additionally, measures must be taken to restrict the influx of antibiotics residue into the environment.

Risk assessment, fitness cost and transcriptome analysis of cyantraniliprole resistance in Spodoptera frugiperda

Spodoptera frugiperda is a notorious invasive pest causing substantial yield losses of crops and has developed resistance to various types of insecticides. In this study, a cyantraniliprole-resistant strain, SfCYAN-R, was obtained from a susceptible strain, SfCYAN-S, after 13 generations of selection with cyantraniliprole. The fitness cost in SfCYAN-R strain was evaluated, and the putative resistance-related genes were explored by RNA-seq analysis. The results showed that SfCYAN-R strain developed 23.97-fold resistance to cyantraniliprole with the realistic heritability of 0.127. The development time of eggs, larvae, prepupae and pupae in SfCYAN-R strain was significantly prolonged than that in SfCYAN-S strain, but no difference in pupation rate, emergence rate and female fecundity was observed between SfCYAN-R and SfCYAN-S strains. Comparative gene expression analysis between SfCYAN-R and SfCYAN-S strains identified 776 significant differentially expressed genes (DEGs), among which several DEGs associated with xenobiotic metabolism were upregulated in SfCYAN-R strain. These results provide insights into the resistance mechanisms of cyantraniliprole and would be helpful for resistance management of S. frugiperda.

Baseline of susceptibility, risk assessment, biochemical mechanism, and fitness cost of resistance to dimpropyridaz, a novel pyridazine pyrazolecarboxamide insecticide, in Bemisia tabaci from China

Bemisia tabaci is one of the most destructive agricultural insect pests around the world, and it has developed high levels of resistance to most pesticides. Dimpropyridaz, a novel insecticide developed by BASF, displays excellent activity against piercing-sucking insect pests. In this study, baseline of susceptibility showed all tested field populations of B. tabaci are susceptible to dimpropyridaz. After continuous selection with dimpropyridaz in the lab, a B. tabaci strain (F12) developed 2.2-fold higher level of resistance compared with a susceptible MED-S strain, and the realized heritability (h2) was estimated as 0.0518. The F12 strain displayed little cross-resistance to afidopyropen, cyantraniliprole, sulfoxaflor, or abamectin, and significantly increased activity of cytochrome P450 monooxygenase (P450). The fitness cost of dimpropyridaz resistance was evident in F12 strain, which had a relative fitness of 0.95 and significantly lower fecundity per female compared with MED-S strain. Taken together, B. tabaci displays high susceptibility to dimpropyridaz in the field, and low risk of developing resistance to dimpropyridaz under successive selection pressure. Little cross-resistance to popular insecticides was found, and fitness cost associated dimpropyridaz resistance was observed. Higher activity of cytochrome P450 in the F12 strain, may be involved in the process of detoxifying dimpropyridaz in whitefly.

Rapid determination and risk evaluation of multi-class antibiotics in aquatic products by one-step purification process coupled with ultra-high performance liquid chromatography–tandem mass spectrometry

A sensitive and robust multiclass analytical method was established to simultaneously determine 55 antibiotics in aquatic products through liquid chromatography–tandem mass spectrometry. A simple one-step purification process was successfully developed, which combined post-acidic acetonitrile extraction directly by an enhanced matrix removal cartridge. This approach eliminated the need for solvent transition. The established method for 55 antibiotics achieved an excellent linear relationship with R2 values ≥ 0.9921 in the range of 0.05–200 μg/L. The quantitation limits ranged within 0.04–5.0 μg/kg. Satisfactory recoveries (76.2%–99.7 %) were achieved with the relative standard deviations below 13.9 %. Furthermore, the antibiotic residues in aquatic products were analyzed, and the health and antibiotic resistance risk assessments were conducted. Although the health risks of target antibiotics were acceptable, a resistance risk was observed. Therefore, monitoring antibiotic residue levels in aquatic products requires considerable attention and further research to ensure the quality of marine products and consumer safety.

Risk assessment, cross-resistance, biochemical mechanism and fitness cost of tetraniliprole resistance in the tomato pinworm Tutaabsoluta

Tuta absoluta is one of the most devastating and invasive insect pests throughout the world. It feeds on numerous solanaceous plant species and has evolved resistance to various types of popular chemical agents. Tetraniliprole is a novel diamide chemical agent that acts as a modulator of the ryanodine receptor, and it has been considered as a promising insecticide against T. absoluta. In this study, we subjected a T. absoluta strain to 17 generations of continuous selection with tetraniliprole in the lab. The resulting tetraniliprole-selected T. absoluta strain (YN-Tet) exhibited a 12.9-fold higher resistance to tetraniliprole compared with a susceptible strain (YN–S), and the realized heritability (h2) was estimated to be 0.1437. The YN-Tet strain exhibited a low level of cross-resistance to chlorantraniliprole and significantly higher cytochrome P450 monooxygenase activity. There was a fitness cost of tetraniliprole resistance in the YN-Tet strain, which had a relative fitness of 0.83 compared with the YN-S strain and a significantly lower fecundity per female. These results enhance our knowledge of the risk and mechanism of tetraniliprole resistance, enabling future optimization of resistance management strategies.

Protective effects of antibiotic resistant bacteria on susceptibles in biofilm: Influential factors, mechanism, and modeling

In environmental biofilms, antibiotic-resistant bacteria facilitate the persistence of susceptible counterparts under antibiotic stresses, contributing to increased community-level resistance. However, there is a lack of quantitative understanding of this protective effect and its influential factors, hindering accurate risk assessment of biofilm resistance in diverse environment. This study isolated an opportunistic Escherichia coli pathogen from soil, and engineered it with plasmids conferring antibiotic resistance. Protective effects of the ampicillin resistant strain (AmpR) on their susceptible counterparts (AmpS) were observed in ampicillin-stress colony biofilms. The concentration of ampicillin delineated protective effects into 3 zones: continuous protection (<1 MIC of AmpS), initial AmpS/R dependent (1–8 MIC of AmpS), and ineffective (>8 MIC of AmpS). Intriguingly, Zone 2 exhibited a surprising “less is more” phenomenon tuned by the initial AmpS/R ratio, where biofilm with an initially lower AmpR (1:50 vs 50:1) harbored 30–90 % more AmpR after 24 h growth under antibiotic stress. Compared to AmpS, AmpR displayed superiority in adhesion, antibiotic degradation, motility, and quorum sensing, allowing them to preferentially colonize biofilm edge and areas with higher ampicillin. An agent-based model incorporating protective effects successfully simulated tempo-spatial dynamics of AmpR and AmpS influenced by antibiotic stress and initial AmpS/R. This study provides a holistic view on the pervasive but poorly understood protective effects in biofilm, enabling development of better risk assessment and precisely targeted control strategies of biofilm resistance in diverse environment.

Resistance risk assessment for benzovindiflupyr in Sclerotium rolfsii and transmission of resistance genes among population.

Sclerotium rolfsii is a destructive soil-borne fungal pathogen which is distributed worldwide. In previous study, the succinate dehydrogenase inhibitor (SDHI) fungicide benzovindiflupyr has been identified for its great antifungal activity against Sclerotium rolfsii. This study is aimed to investigate the resistance risk and mechanism of benzovindiflupyr in Sclerotium rolfsii. Eight stable benzovindiflupyr-resistant isolates were generated by fungicide adaptation. Although the obtained eight resistant isolates have a stronger pathogenicity than the parental sensitive isolate, they have a fitness penalty in the mycelial growth and sclerotia formation compared to the parental isolate. A positive cross-resistance existed in the resistant isolates between benzovindiflupyr and thifluzamide, carboxin, boscalid and isopyrazam. Three-point mutations, including SdhBN180D, SdhCQ68E and SdhDH103Y, were identified in the benzovindiflupyr-resistant isolates. However, molecular docking analysis indicated that only SdhDH103Y could influence the sensitivity of Sclerotium rolfsii to benzovindiflupyr. After mycelial co-incubation of resistant isolates and the sensitive isolate, resistance genes may be transmitted to the sensitive isolate. The in vivo efficacy of benzovindiflupyr and thifluzamide against benzovindiflupyr-resistant isolates was a little lower than that against the sensitive isolate but with no significant difference. The results suggested a low to medium resistance risk of Sclerotium rolfsii to benzovindiflupyr. However, once resistance occurs, it is possible to spread in the population of Sclerotium rolfsii. This study is helpful to understanding the risk and mechanism of resistance to benzovindiflupyr in multinucleate pathogens such as Sclerotium rolfsii. © 2024 Society of Chemical Industry.

Health risk assessment and characterization of PM2.5 bound bioaerosols at the municipal solid waste landfill site of Nagpur, India

Landfill being the reservoir of organic waste, serves as the most important source in the generation and emission of bioaerosols in the environment, and these bioaerosols have a significant impact on the environment and public health. In India, there are limited studies on the characterization of culturable bacteria and fungi in the bioaerosols originating from landfills. Therefore, the present study aimed to quantify and identify the bioaerosols and study their characteristics at a landfill site in Nagpur, India. The study describes the seasonal variation of PM2.5, PM2.5-associated bacteria and fungi in the landfill air, their molecular identification, antibiotic resistance, pathogenicity, dispersion and health risk assessment during winter and pre-monsoon seasons at the landfill site and surrounding residential sites in upwind and downwind directions. The study showed PM2.5 concentration was highest in the winter season (265.56 μg/m3) followed by the pre-monsoon season (173.1 μg/m3) at the landfill site. The culturable bioaerosol level was also significantly higher in winter (11056 CFU/m3) in comparison to pre-monsoon season (2244 CFU/m3) at the landfill site. Upon isolating and characterizing the microorganisms in the bioaerosols, some of the bacterial isolates, like Bacillus, Staphylococcus gallinarum, and Streptomyces speibonae were found to be resistant to chloramphenicol, netillin, nitrofurantoin, streptomycin etc. About 30% of the bacterial isolates were found to show β-hemolytic virulence activity. This indicated the presence of multi-drug resistant and potentially pathogenic bacteria in the bioaerosols over the landfill. The health risk assessment in adults indicated that the workers at the landfill site were at risk of bacterial aerosol in the winter season (hazard quotient i.e., HQ>1). The impact of bioaerosols on children needs to be investigated in the further studies and regular monitoring of bioaerosols is suggested to avoid any morbidity. To the best of our knowledge, this is the first study on the characterization of microorganisms from the bioaerosols over an open landfill site in India.

Resistance risk assessment, cross-resistance potential and realized heritability of resistance to methomyl in Musca domestica Linnaeus.

The use of insecticides in agricultural settings often exerts negative effects on nontarget species. Methomyl, a broad-spectrum carbamate insecticide, is recommended to manage a number of insect pests of the cotton crop. Recently, Musca domestica, which is a nontarget insect species in cotton fields, has shown resistance to methomyl in Pakistan. The present study tried to assess resistance-risk assessment, rapidity of resistance development to methomyl, cross-resistance potential to other insecticides, resistance heritability and to forecast the projected rate of resistance development under field conditions. For this purpose, a field strain of M. domestica with 186 fold resistance to methomyl was re-selected in the laboratory for eight consecutive generations. Consequently, LD50 values increased rapidly (126.64 ng/fly to 3112.79 ng/fly) compared to those before selection experiments. Similarly, RR values increased from 186 to 3113 fold as a result of the selection process. However, resistance to methomyl did not remain stable when the selected strain (Meth-SEL) reared for the next five generations in a pesticide free environment. The Meth-SEL strain also developed cross-resistance to permethrin. The realized heritability (h2) value for the Meth-SEL strain was 0.39 with 27% average mortality of M. domestica. Assuming the standard deviation (σp) value 0.27 and the h2 value 0.39 for eight generations of continuous exposure to methomyl, then five, seven, eight, ten and twelve generations at 90, 80, 70, 60 and 50% selection intensity, respectively, would be required for a tenfold increase in the LD50 value of methomyl. In conclusion, the Meth-SEL strain of M. domestica exhibited a high risk of resistance development to methomyl under continuous selection pressure. Resistance increased rapidly during selection experiments that reflect the probability of resistance development under field conditions if M. domestica receive exposures to methomyl during its applications for the management of target pest species.

Risk assessment and molecular mechanism of Sclerotium rolfsii resistance to boscalid

Boscalid, a widely used SDHI fungicide, has been employed in plant disease control for over two decades. However, there is currently no available information regarding its antifungal activity against Sclerotium rolfsii and the potential risk of resistance development in this pathogen. In this study, we evaluated the sensitivity of 100 S. rolfsii strains collected from five different regions in China during 2018–2019 to boscalid using mycelial growth inhibition method and assessed the risk of resistance development. The EC50 values for boscalid ranged from 0.2994 μg/mL to 1.0766 μg/mL against the tested strains, with an average EC50 value of 0.7052 ± 0.1473 μg/mL. Notably, a single peak sensitivity baseline was curved, indicating the absence of any detected resistant strains. Furtherly, 10 randomly selected strains of S. rolfsii were subjected to chemical taming to evaluate its resistance risk to boscalid, resulting in the successful generation of six stable and inheritable resistant mutants. These mutants exhibited significantly reduced mycelial growth, sclerotia production, and virulence compared to their respective parental strains. Cross-resistance tests revealed a correlation between boscalid and flutolanil, benzovindiflupyr, pydiflumetofen, fluindapyr, and thifluzamide; however, no cross-resistance was observed between boscalid and azoxystrobin. Thus, we conclude that the development risk of resistance in S. rolfsii to boscalid is low. Boscalid can be used as an alternative fungicide for controlling peanut sclerotium blight when combined with other fungicides that have different mechanisms of action. Finally, the target genes SDHB, SDHC, and SDHD in S. rolfsii were initially identified, cloned and sequenced to elucidate the mechanism of S. rolfsii resistance to boscalid. Two mutation genotypes were found in the mutants: SDHD-D111H and SDHD-H121Y. The mutants carrying SDHD-H121Y exhibited moderate resistance, while the mutants with SDHD-D111H showed low resistance. These findings contribute to our comprehensive understanding of molecular mechanisms underlying plant pathogens resistance to SDHI fungicides.

Prediction of antibiotic sorption in soil with machine learning and analysis of global antibiotic resistance risk

Although the sorption of antibiotics in soil has been extensively studied, their spatial distribution patterns and sorption mechanisms still need to be clarified, which hinders the assessment of antibiotic resistance risk. In this study, machine learning was employed to develop the models for predicting the soil sorption behavior of three classes of antibiotics (sulfonamides, tetracyclines, and fluoroquinolones) in 255 soils with 2203 data points. The optimal independent models obtained an accurate predictive performance with R2 of 0.942 to 0.977 and RMSE of 0.051 to 0.210 on test sets compared to combined models. Besides, a global map of the antibiotic sorption capacity of soil predicted with the optimal models revealed that the sorption potential of fluoroquinolones was the highest, followed by tetracyclines and sulfonamides. Additionally, 14.3% of regions had higher antibiotic sorption potential, mainly in East and South Asia, Central Siberia, Western Europe, South America, and Central North America. Moreover, a risk index calculated with the antibiotic sorption capacity of soil and population density indicated that about 3.6% of soils worldwide have a high risk of resistance, especially in South and East Asia with high population densities. This work has significant implications for assessing the antibiotic contamination potential and resistance risk.