Antimicrobial resistance in Bacillus-based biopesticide products

The crisis of antimicrobial resistant bacterial infections is one of the most pressing public health issues. Common agricultural practices have been implicated in the generation of antimicrobial resistant bacteria. Biopesticides, live bacteria used for pest control, are non-pathogenic and considered safe for consumption. Application of bacteria-based pesticides to crops in high concentrations raises the possibility of unintentional contributions to the movement and generation of antimicrobial resistance genes in the environment. However, the presence of clinically relevant antimicrobial resistance genes and their resistance phenotypes are currently unknown. Here we use a combination of multiple bioinformatic and microbiological techniques to define resistomes of widely used biopesticides and determine how the presence of suspected antimicrobial resistance genes translates to observable resistance phenotypes in several biopesticide products. Our results demonstrate that biopesticide products are reservoirs of clinically relevant antimicrobial resistance genes and bear resistance to multiple drug classes.

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Antibiotic Resistance in Bacillus-based Biopesticide Products

10.1101/2021.03.15.435560 ◽

2021 ◽

Author(s):

Mo Kaze ◽

Mark Sistrom ◽

Lauren Brooks

Keyword(s):

Antibiotic Resistance ◽

Resistance Genes ◽

Bacterial Infections ◽

Antibiotic Resistance Genes ◽

Agricultural Practices ◽

Resistant Bacteria ◽

Multiple Drug ◽

Antibiotic Resistant ◽

Microbiological Techniques ◽

High Concentrations

The crisis of antibiotic resistant bacterial infections is one of the most pressing public health issues. Common agricultural practices have been implicated in the generation of antibiotic resistant bacteria. Biopesticides, live bacteria used for pest control, are non-pathogenic and considered safe for consumption. Application of bacteria-based pesticides to crops in high concentrations raises the possibility of unintentional contributions to the movement and generation of antibiotic resistance genes in the environment. However, the presence of clinically relevant antibiotic resistance genes and their resistance phenotypes are currently unknown. Here we use a combination of multiple bioinformatic and microbiological techniques to define resistomes of widely used biopesticides and determine how the presence of suspected antibiotic resistance genes translates to observable resistance phenotypes in several biopesticide products. Our results demonstrate that biopesticide products are reservoirs of clinically relevant antibiotic resistance genes and bear resistance to multiple drug classes.

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Multi-omic surveillance of Escherichia coli and Klebsiella spp. in hospital sink drains and patients

10.1101/2020.02.19.952366 ◽

2020 ◽

Author(s):

B Constantinides ◽

KK Chau ◽

TP Quan ◽

G Rodger ◽

M Andersson ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Bacterial Infections ◽

Wide Spectrum ◽

Klebsiella Oxytoca ◽

Clinical Disease ◽

E Coli ◽

Antimicrobial Resistance Genes ◽

Wide Range

ABSTRACTEscherichia coli and Klebsiella spp. are important human pathogens that cause a wide spectrum of clinical disease. In healthcare settings, sinks and other wastewater sites have been shown to be reservoirs of antimicrobial-resistant E. coli and Klebsiella spp., particularly in the context of outbreaks of resistant strains amongst patients. Without focusing exclusively on resistance markers or a clinical outbreak, we demonstrate that many hospital sink drains are abundantly and persistently colonised with diverse populations of E. coli, Klebsiella pneumoniae and Klebsiella oxytoca, including both antimicrobial-resistant and susceptible strains. Using whole genome sequencing (WGS) of 439 isolates, we show that environmental bacterial populations are largely structured by ward and sink, with only a handful of lineages, such as E. coli ST635, being widely distributed, suggesting different prevailing ecologies which may vary as a result of different inputs and selection pressures. WGS of 46 contemporaneous patient isolates identified one (2%; 95% CI 0.05-11%) E. coli urine infection-associated isolate with high similarity to a prior sink isolate, suggesting that sinks may contribute to up to 10% of infections caused by these organisms in patients on the ward over the same timeframe. Using metagenomics from 20 sink-timepoints, we show that sinks also harbour many clinically relevant antimicrobial resistance genes including blaCTX-M, blaSHV and mcr, and may act as niches for the exchange and amplification of these genes. Our study reinforces the potential role of sinks in contributing to Enterobacterales infection and antimicrobial resistance in hospital patients, something that could be amenable to intervention.IMPORTANCEEscherichia coli and Klebsiella spp. cause a wide range of bacterial infections, including bloodstream, urine and lung infections. Previous studies have shown that sink drains in hospitals may be part of transmission chains in outbreaks of antimicrobial-resistant E. coli and Klebsiella spp., leading to colonisation and clinical disease in patients. We show that even in non-outbreak settings, contamination of sink drains by these bacteria is common across hospital wards, and that many antimicrobial resistance genes can be found and potentially exchanged in these sink drain sites. Our findings demonstrate that the colonisation of handwashing sink drains by these bacteria in hospitals is likely contributing to some infections in patients, and that additional work is needed to further quantify this risk, and to consider appropriate mitigating interventions.

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Merging Metagenomics and Spatial Epidemiology To Understand the Distribution of Antimicrobial Resistance Genes from Enterobacteriaceae in Wild Owls

Applied and Environmental Microbiology ◽

10.1128/aem.00571-20 ◽

2020 ◽

Vol 86 (20) ◽

Author(s):

Elizabeth A. Miller ◽

Julia B. Ponder ◽

Michelle Willette ◽

Timothy J. Johnson ◽

Kimberly L. VanderWaal

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

Spatial Epidemiology ◽

Anthropogenic Sources ◽

Resistant Bacteria ◽

Metagenomic Sequencing ◽

Wild Animals ◽

Natural Ecosystems ◽

Content Type ◽

Antimicrobial Resistance Genes

ABSTRACT Antimicrobial resistance (AMR) is a well-documented phenomenon in bacteria from many natural ecosystems, including wild animals. However, the specific determinants and spatial distribution of resistant bacteria and antimicrobial resistance genes (ARGs) in the environment remain incompletely understood. In particular, information regarding the importance of anthropogenic sources of AMR relative to that of other biological and ecological influences is lacking. We conducted a cross-sectional study of AMR in great horned owls (Bubo virginianus) and barred owls (Strix varia) admitted to a rehabilitation center in the midwestern United States. A combination of selective culture enrichment and shotgun metagenomic sequencing was used to identify ARGs from Enterobacteriaceae. Overall, the prevalence of AMR was comparable to that in past studies of resistant Enterobacteriaceae in raptors, with acquired ARGs being identified in 23% of samples. Multimodel regression analyses identified seasonality and owl age to be important predictors of the likelihood of the presence of ARGs, with birds sampled during warmer months being more likely to harbor ARGs than those sampled during cooler months and with birds in their hatch year being more likely to harbor β-lactam ARGs than adults. Beyond host-specific determinants, ARG-positive owls were also more likely to be recovered from areas of high agricultural land cover. Spatial clustering analyses identified a significant high-risk cluster of tetracycline resistance gene-positive owls in the southern sampling range, but this could not be explained by any predictor variables. Taken together, these results highlight the complex distribution of AMR in natural environments and suggest that both biological and anthropogenic factors play important roles in determining the emergence and persistence of AMR in wildlife. IMPORTANCE Antimicrobial resistance (AMR) is a multifaceted problem that poses a worldwide threat to human and animal health. Recent reports suggest that wildlife may play an important role in the emergence, dissemination, and persistence of AMR. As such, there have been calls for better integration of wildlife into current research on AMR, including the use of wild animals as biosentinels of AMR contamination in the environment. A One Health approach can be used to gain a better understanding of all AMR sources and pathways, particularly those at the human-animal-environment interface. Our study focuses on this interface in order to assess the effect of human-impacted landscapes on AMR in a wild animal. This work highlights the value of wildlife rehabilitation centers for environmental AMR surveillance and demonstrates how metagenomic sequencing within a spatial epidemiology framework can be used to address questions surrounding AMR complexity in natural ecosystems.

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Research and Technological Advances Regarding the Study of the Spread of Antimicrobial Resistance Genes and Antimicrobial-Resistant Bacteria Related to Animal Husbandry

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph16244896 ◽

2019 ◽

Vol 16 (24) ◽

pp. 4896

Author(s):

Na Li ◽

Chong Liu ◽

Zhiguo Zhang ◽

Hongna Li ◽

Tingting Song ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

High Throughput Sequencing ◽

Animal Husbandry ◽

Control Measures ◽

Research Progress ◽

Future Research ◽

Resistant Bacteria ◽

Technological Advances ◽

Antimicrobial Resistance Genes

The extensive use of antimicrobials in animal farms poses serious safety hazards to both the environment and public health, and this trend is likely to continue. Antimicrobial resistance genes (ARGs) are a class of emerging pollutants that are difficult to remove once introduced. Understanding the environmental transfer of antimicrobial-resistant bacteria (ARB) and ARGs is pivotal for creating control measures. In this review, we summarize the research progress on the spread and detection of ARB and ARG pollution related to animal husbandry. Molecular methods such as high-throughput sequencing have greatly enriched the information about ARB communities. However, it remains challenging to delineate mechanisms regarding ARG induction, transmission, and tempo-spatial changes in the whole process, from animal husbandry to multiple ecosystems. As a result, future research should be more focused on the mechanisms of ARG induction, transmission, and control. We also expect that future research will rely more heavily on metagenomic -analysis, metatranscriptomic sequencing, and multi-omics technologies

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Characterization and Horizontal Transfer of Antimicrobial Resistance Genes and Integrons in Bacteria Isolated from Cooked Meat Products in China

Journal of Food Protection ◽

10.4315/0362-028x.jfp-17-119 ◽

2017 ◽

Vol 80 (12) ◽

pp. 2048-2055 ◽

Cited By ~ 2

Author(s):

Tao Yu ◽

Xiaobing Jiang ◽

Yu Liang ◽

Yanping Zhu ◽

Jinhe Tian ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

Meat Products ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Integrase Gene ◽

Antimicrobial Resistance Genes ◽

Class 1 ◽

Cooked Meat ◽

Cooked Meat Products

ABSTRACT The aim of this study was to investigate antimicrobial resistance and the presence and transferability of corresponding resistance genes and integrons in bacteria isolated from cooked meat samples in the People's Republic of China. A total of 150 isolates (22 species belonging to 15 genera) were isolated from 49 samples. Resistance of these isolates to antimicrobials was commonly observed; 42.7, 36.0, and 25.3% of the isolates were resistant to tetracycline, streptomycin, and ampicillin, respectively. Multidrug resistance was observed in 41 (27.3%) of the isolates. Sixteen resistance genes, i.e., blaTEM-1 and blaCTX-M-14 (β-lactams), aac(3)-IIa (gentamicin), strA and strB (streptomycin), qnrB and qnrS (fluoroquinolone), sul1, sul2, and sul3 (sulfamethoxazole), cat1 and cat2 (chloramphenicol), and tetM, tetA, tetS, and tetB (tetracycline), were found in 54 isolates. One isolate of Pseudomonas putida carried qnrB, and sequence analysis of the PCR product revealed 96% identity to qnrB2. The qnr genes were found coresiding and were cotransferred with bla genes in two isolates. Twelve isolates were positive for the class 1 integrase gene, and four isolates carried the class 2 integrase gene. However, no class 3 integrase gene was detected. One isolate of Proteus mirabilis carried dfrA32-ereA-aadA2, and this unusual array could be transferred to Escherichia coli. Nonclassic class 1 integrons lacking qacEΔ1 and sul1 genes were found in 2 of the 12 intI1-positive isolates. Our results revealed the presence of multidrug-resistant bacteria in cooked meats and the presence and transferability of resistance genes in some isolates, suggesting that cooked meat products may act as reservoirs of drug-resistant bacteria and may facilitate the spread of resistance genes.

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Untreated Liquid Hospital Waste: Potential Source of Multidrug Resistant Bacteria

Bangladesh Journal of Microbiology ◽

10.3329/bjm.v32i0.28473 ◽

2016 ◽

pp. 21-24

Author(s):

Md Kamruzzaman Siddiqui ◽

Nazma Khatoon ◽

Pravas Chandra Roy

Keyword(s):

Antimicrobial Resistance ◽

Waste Water Treatment Plant ◽

Diffusion Method ◽

Resistant Bacteria ◽

Multiple Drug ◽

Drug Resistant ◽

Salmonella Spp ◽

Antibiotic Resistant ◽

E Coli ◽

Antimicrobial Resistance Genes

Antimicrobial resistance in both pathogenic and commensal bacteria is increasing steadily. Failure of antibiotic resistant bacteria containment is responsible for this expansion. Healthcare effluent acts as the store house of harmful infectious pathogens. Potential health risk includes spreading of diseases by these pathogens and wide dissemination of antimicrobial resistance genes. The present study was carried out to investigate the multiple-drug resistance among the bacterial strains that were isolated and identified from the effluents of Jessore Medical College Hospital & Jessore Queens hospital private limited. Identified bacteria were E. coli , Klebsiella spp., Enterobacter spp., Proteus vulgaris and Salmonella spp.. Occurrence of E. coli and Enterobacter spp. were found to have the highest percentages and present in majority of the samples. The identified organisms antibiotic resistant pattern were analyzed by agar disc diffusion method against 6 antibiotics. Results of antibiotic susceptibility test showed that all of the isolates were multi-drug resistant (e4). From the study, we observed that 75% of the isolates were resistant to amoxicillin, followed by Ampicillin (64%), Chloramphenicol (31%), Gentamycin (29%), Nitrofurantoin (27%) and least resistant being Ciprofloxacin 23%. Among the isolates Salmonella spp. were showed highest rate of resistance against all the used antibiotics. The result denotes that, the identified bacteria have been well exposed to the tested antimicrobials and they have established mechanisms to avoid them. Therefore, proper waste water treatment plant should be established to diminish the risk of disseminating multiple drug resistant microorganisms for the safeguard of public health.Bangladesh J Microbiol, Volume 32, Number 1-2,June-Dec 2015, pp 21-24

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Prevalence of virulence and antimicrobial resistance genes in Salmonella spp. isolated from commercial chickens and human clinical isolates from South Africa and Brazil

Onderstepoort Journal of Veterinary Research ◽

10.4102/ojvr.v83i1.1067 ◽

2016 ◽

Vol 83 (1) ◽

Cited By ~ 27

Author(s):

Oliver T. Zishiri ◽

Nelisiwe Mkhize ◽

Samson Mukaratirwa

Keyword(s):

Drug Resistance ◽

Antimicrobial Resistance ◽

South African ◽

Resistance Genes ◽

Broiler Chickens ◽

Antimicrobial Agents ◽

Multiple Drug Resistance ◽

Multiple Drug ◽

Poultry Production ◽

Antimicrobial Resistance Genes

Salmonellosis is a significant public health concern around the world. The injudicious use of antimicrobial agents in poultry production for treatment, growth promotion and prophylaxis has resulted in the emergence of drug resistant strains of Salmonella. The current study was conducted to investigate the prevalence of virulence and antimicrobial resistance genes from Salmonella isolated from South African and Brazilian broiler chickens as well as human clinical isolates. Out of a total of 200 chicken samples that were collected from South Africa 102 (51%) tested positive for Salmonella using the InvA gene. Of the overall 146 Salmonella positive samples that were screened for the iroB gene most of them were confirmed to be Salmonella enterica with the following prevalence rates: 85% of human clinical samples, 68.6% of South African chicken isolates and 70.8% of Brazilian chicken samples. All Salmonella isolates obtained were subjected to antimicrobial susceptibility testing with 10 antibiotics. Salmonella isolates from South African chickens exhibited resistance to almost all antimicrobial agents used, such as tetracycline (93%), trimethoprim-sulfamthoxazole (84%), trimethoprim (78.4%), kanamycin (74%), gentamicin (48%), ampicillin (47%), amoxicillin (31%), chloramphenicol (31%), erythromycin (18%) and streptomycin (12%). All samples were further subjected to PCR in order to screen some common antimicrobial and virulence genes of interest namely spiC, pipD, misL, orfL, pse-1, tet A, tet B, ant (3")-la, sul 1 and sul. All Salmonella positive isolates exhibited resistance to at least one antimicrobial agent; however, antimicrobial resistance patterns demonstrated that multiple drug resistance was prevalent. The findings provide evidence that broiler chickens are colonised by pathogenic Salmonella harbouring antimicrobial resistance genes. Therefore, it is evident that there is a need for prudent use of antimicrobial agents in poultry production systems in order to mitigate the proliferation of multiple drug resistance across species.Keywords: Salmonella; antimicrobial resistance; chicken; human; susceptibility; virulence gene

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Antibiotic Resistance Profile and Resistance Determination of Bacteria Isolated from Water in Southern Benin

Journal of Advances in Microbiology ◽

10.9734/jamb/2021/v21i430345 ◽

2021 ◽

pp. 91-105

Author(s):

Hornel Koudokpon ◽

Victorien Dougnon ◽

Christelle Lougbegnon ◽

Esther Deguenon ◽

Wassiyath Mousse ◽

...

Keyword(s):

Drinking Water ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Water Samples ◽

Meca Gene ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Gram Negative ◽

Antimicrobial Resistance Genes ◽

Southern Benin

Background: The environment plays an important role in the dissemination of multidrug resistant bacteria, especially through the aquatic ecosystem, including hospital effluents, rivers, but also spring water and drinking water. This study aims to determine selected antimicrobial resistance genes in some aquatic matrices in southern Benin. Methods: Collected water samples were filtered through a membrane 0.22 µm thick. After filtration, the membrane was deposited on Muëller Hinton agar. Then the colonies resulting from this subculture were subjected to a microbiological examination by the conventional method. The antibiotic sensitivity test was carried out by the Kirby Bauer method according to the recommendations of the French Society of Microbiology. Resistance genes were looked for by PCR. Results: Of the 222 water samples collected, 265 bacterial strains were isolated, the majority of which were strains of Coagulase Negative Staphylococcus (CNS) with 37.74% (n = 100), followed by strains of Klebsiella pneumoniae (21.89%; n = 58), Escherichia coli (10.57%; n = 28). All isolated gram-negative bacilli strains are multidrug resistant with resistance of all strains to amoxicillin, ampicillin and amoxicillin + clavulanic acid. Of the 15 resistance genes searched in the genome of Gram-negative bacilli strains, 8 were detected, namely the TEM, SHV, CTX-M15, VIM, NDM, SUL1, SUL2 and AADA genes. Resistance of CNS strains to amoxicillin, oxacillin and cefoxitin was observed. The meca gene was detected in all CNS strains. The vanA and VanB genes were only detected in strains isolated from drinking water in sachets collected from producers and street sellers. Conclusion: These results show the dissemination of resistance genes in Benin and once again confirms the urgency of a global fight against antimicrobial resistance.

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