Bacteriological quality of fresh and processed black soldier fly Hermetia illucens larvae reared on chicken manure in Kitwe, Zambia. - Related Documents




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257901.0000Bacteriological quality of fresh and processed black soldier fly Hermetia illucens larvae reared on chicken manure in Kitwe, Zambia. Black soldier fly larvae (BSFL) have high nutrient content and are rapidly becoming an alternative protein source for animal feed. However, microbial contamination is a potential risk due to the environment in which they are reared. This study assessed the bacteriological quality of fresh and processed BSFL by comparing the processed BSFL using two traditional methods (oven-dried and sun-dried), on their effectiveness at reducing the bacterial load and further elucidated bacterial composition. PCR was used to identify extended-spectrum β-lactamase (ESBL) and mecA genes in Escherichia coli and Staphylococcus spp., respectively. A total of 51 fresh BSFL samples were collected from a commercial poultry farm in Kitwe, Zambia. The results showed various bacterial genera, with a higher diversity among gram-positive isolates. The comparison of the effectiveness of two traditional processing methods for BSFL, sun-drying and oven-drying, showed that both methods significantly reduced the bacterial load, with oven-drying causing a larger reduction. While various genera were identified, we focused on E. coli and Staphylococcus spp. This is because some E. coli harbor ESBLs that hydrolyze β-lactam antibiotics like cephalosporins and penicillin, leading to resistance. Similarly, the genus Staphylococcus was selected since some strains are potentially pathogenic and contain the mecA gene that encodes resistance to β-lactam antibiotics. Molecular characterization of the isolated strains revealed bla(CTX-M) and bla(TEM) genes among E. coli, but the mecA gene was not detected among Staphylococcus. This study revealed that BSFL harbor bacteria of zoonotic significance, emphasizing the need for good processing methods to eliminate potential risks. IMPORTANCE: Isolation and identification of Escherichia coli and Staphylococcus spp. in processed black soldier fly larvae (BSFL) samples meant for animal feed indicate insufficient processing methods and pose a public health risk. For instance, some E. coli harbor extended-spectrum β-lactamases (ESBLs) that hydrolyze β-lactam antibiotics like cephalosporins and penicillin, leading to resistance. In addition, some E. coli commensals can transfer antimicrobial resistance genes to pathogenic bacteria through horizontal gene transfer using various mobile genetic elements, leading to resistance. Similarly, for Staphylococcus spp., some strains of the genus Staphylococcus are potentially pathogenic and contain the mecA gene that encodes resistance to β-lactam antibiotics. In this study, we used PCR to screen E. coli isolates for the two commonly reported ESBL genes in Zambia, bla(CTX-M) and bla(TEM), and Sanger sequencing was used to reveal bla(CTX-M) gene alleles. Our results highlight the importance of using adequate processing methods for BSFL to eliminate potential health risks to animal feed.202540539803
192910.9997Research Note: Detection of antibiotic-resistance genes in commercial poultry and turkey flocks from Italy. Antibiotics are routinely used in commercial poultry farms for the treatment of economically important bacterial diseases. Repeated use of antibiotics, usually administered in the feed or drinking water, may also result in the selection of resistant bacteria in animal feces, able to transfer their antimicrobial-resistance genes (ARG), residing on mobile elements, to other microorganisms, including human pathogens. In this study, single and multiplex PCR protocols were performed to detect tetracycline-, lincomycin-, chloramphenicol-, aminoglycoside-, colistin-, vancomycin-, and carbapenem-resistance genes, starting from 38 litter samples collected from 6 poultry and 2 turkey Italian flocks. The ARG were confirmed for all investigated classes of antimicrobials, except for colistin (mcr-1, mcr-2, mcr-3,mcr-4 mcr-5) and carbapenem (IMP, OXA-48, NDM, KPC), while the vanB gene was only detected for vancomycin. The highest positivity was obtained for tetracycline (tet[L], tet[M], tet[K], tetA[P]] and aminoglycoside (aadA2) ARG, confirming the predominant use of these antimicrobials in the veterinary practice and their potential to enhance the resistance patterns also in humans as a consequence of environmental contamination. On the contrary, the dissemination by poultry of ARG for critically important antimicrobials seems to be of minor concern, suggesting a negligible environmental dissemination by these genes in the Italian poultry industry. Finally, the molecular screening performed in this study using a noninvasive sampling method represents a simple and rapid tool for monitoring the ARG patterns at the farm level.202133799114
557020.9997Monitoring the Spread of Multidrug-Resistant Escherichia coli Throughout the Broiler Production Cycle. The extensive use of antimicrobials in broiler production is changing the bird microbiota, fostering drug-resistant bacteria, and complicating therapeutic interventions, making the problem of multidrug resistance global. The monitoring of antimicrobial virulence and resistance genes are tools that have come to assist the breeding of these animals, directing possible treatments as already used in human medicine and collecting data to demonstrate possible dissemination of multidrug-resistant strains that may cause damage to industry and public health. This work aimed to monitor broiler farms in southern Brazil, isolating samples of E. coli and classifying them according to the profile of resistance to antimicrobials of interest to human and animal health. We also monitored the profile of virulence genes and conducted an epidemiological survey of possible risk factors that contribute to this selection of multidrug-resistant isolates. Monitoring was carried out on farms in the three southern states of the country, collecting samples of poultry litter, cloacal swabs, and beetles of the species Alphitobius diaperinus, isolating E. coli from each of these samples. These were evaluated by testing their susceptibility to antimicrobials of animal and human interest; detecting whether the samples were extended-spectrum β-lactamase enzyme (ESBL) producers; and when positive, selected for genotypic tests to identify resistant genes (CTX-M, TEM, and SHV) and virulence. Among the antimicrobials tested, enrofloxacin and ciprofloxacin demonstrated some of the highest frequencies of resistance in the isolated strains, with significant statistical results. The use of these antimicrobials increased the likelihood of resistance by over three times and was associated with a 1.5-fold higher probability of multidrug resistance. Of all isolates, 95% were multidrug-resistant, raising concerns for production and public health. Among 231 ESBL-positive samples, the CTX-M1 group predominated.202539858355
192830.9997Targeted Antimicrobial Resistance Gene Screening from Metagenomic DNA of Raw Milk Samples Identifies the Presence of Multiple Genes Including the mcr9. The current study has investigated the prevalence of antimicrobial resistance (AMR) genes in cow and goat raw milk samples. The misuse of antibiotics in the livestock sector has already been reported to be a major factor contributing to AMR risk. For the study, milk samples were collected from five different farms, and metagenomic DNA was extracted. Then, PCR amplification was carried out using primers specific to 15 different AMR genes. From the results obtained, the prevalence of β-lactam resistance genes, particularly blaTEM (24%), along with other genes like blaZ (12%) and blaSHV (8%), were observed in addition to the transmissible mcr9 gene (12%) conferring resistance to colistin. These findings underscore the urgent need for monitoring AMR genes and regulating antibiotic use in dairy farming to safeguard public health, as it poses a potential risk with the consumption of unpasteurized milk.202540488653
193340.9997Antibiotic Resistance Genes Occurrence in Conventional and Antibiotic-Free Poultry Farming, Italy. Antimicrobial resistance is a complex and widespread problem threatening human and animal health. In poultry farms, a wide distribution of resistant bacteria and their relative genes is described worldwide, including in Italy. In this paper, a comparison of resistance gene distribution in litter samples, recovered from four conventional and four antibiotic-free broiler flocks, was performed to highlight any influence of farming systems on the spreading and maintenance of resistance determinants. Conventional PCR tests, targeting the resistance genes related to the most used antibiotics in poultry farming, along with some critically important antibiotics for human medicine, were applied. In conventional farms, n. 10 out of n. 30 investigated genes were present in at least one sample, the most abundant fragments being the tet genes specific for tetracyclines, followed by those for aminoglycosides and chloramphenicol. All conventional samples resulted negative for colistin, carbapenems, and vancomycin resistance genes. A similar trend was observed for antibiotic-free herds, with n. 13 out of n. 30 amplified genes, while a positivity for the mcr-1 gene, specific for colistin, was observed in one antibiotic-free flock. The statistical analysis revealed a significant difference for the tetM gene, which was found more frequently in the antibiotic-free category. The analysis carried out in this study allowed us to obtain new data about the distribution of resistance patterns in the poultry industry in relation to farming types. The PCR test is a quick and non-expensive laboratory tool for the environmental monitoring of resistance determinants identifying potential indicators of AMR dissemination.202236139170
557150.9997ESβL E. coli isolated in pig's chain: Genetic analysis associated to the phenotype and biofilm synthesis evaluation. Resistance to new generation cephalosporins is an important public health problem globally, in terms of economic and social costs, morbidity and mortality. Βeta-lactamase enzymes are mainly responsible for the antibiotic resistance of Gram negative bacteria and extended-spectrum-β-lactamases (ESβLs) are one of the major determinants of resistance against oxymino-cephalosporins in Enterobacteriaceae. Food-producing animals represent one of the sources of antibiotic resistant bacteria, including pigs. Here we analysed the presence of E. coli resistant to III generation cephalosporins isolated from different matrices collected from intensively bred pigs. A total of 498 E. coli were isolated from faeces and carcasses of pigs at slaughterhouse as well as from pork meat and sausages. Among these, 73 were phenotypically confirmed to be ESβL producers. Genetic analysis revealed that all except two harboured at least one of the three selected genes: bla(CTX-M), bla(TEM), and bla(SHV). Furthermore, six of the E. coli ESβL isolated from faeces and carcasses swabs, were also able to produce biofilm, highlighting the virulence potential of these strains. The presence of Multi-Drug-Resistance patterns (MDR) recorded by the 73 ESβL E. coli was significant (60% of the strains were resistant to more than six antibiotics in MIC test). Results from the present study show that the transmission of resistant bacteria is possible along the food chain, including production of pork, one the most highly consumed meats around the world. Transmission is possible through the ingestion of raw meat products, and following cross-contamination between raw and cooked foods during preparation. The potential risk for human health demonstrated here, associated with the consumption of pork contaminated with bacterial strains characterized by multidrug resistance patterns, and the ability to produce ESβL and biofilm, is cause for concern. It is imperative to study future control strategies to avoid or limit as much as possible the transmission of these highly pathogenic strains through food consumption and/or contact with the environment.201930245289
159260.9997Identification of ESBL-Producing Enterobacterales From Vegetable Plants: Preliminary Findings From a Small Cross-Sectional Study in a Rural Area of Madagascar. Extended-spectrum beta-lactamases (ESBL)-producing enterobacterales are considered a key indicator for antimicrobial resistance (AMR) epidemiological surveillance in animal, human, and environment compartments. In this study, we aim to investigate the presence and genetic diversity of ESBL-producing enterobacterales on vegetable plants. We isolated beta-lactam resistant enterobacterales from several vegetable plants and sequenced their whole genome. Utilising standard genomic and phylogenetic methods, we sought to (i) characterise the resistance genes and plasmid content of the plant-isolated strains, (ii) investigate their genetic structure, and (iii) determine their relationships with strains from other reservoirs. Among the 22 strains collected from vegetable plants, 6 showed resistance to beta-lactam antibiotics, with 5 of them identified as ESBL producers. Our results indicated the presence of multidrug-resistant (MDR) strains containing multiple antibiotic resistance genes (ARGs). Importantly, no host-specific lineages were identified among the plant-isolated ESBL-producing E. coli (ESBL-Ec). Instead, these strains exhibited genetic and epidemiological connections with strains isolated from animals, humans, and the environment, suggesting transfer of ESBL-Ec between plants and other sources in rural Madagascar. These preliminary findings suggest that vegetable plants are contaminated as a result of human activities, posing a potential risk of human and animal exposure to antibiotic-resistant bacteria and genes.202540528688
192770.9997First Molecular Characterization and Antibiogram of Bacteria Isolated From Dairy Farm Wastewater in Bangladesh. This pioneering study in Bangladesh combines phenotypic and genotypic approaches to characterize antibiotic-resistant bacteria in dairy farm wastewater, addressing a critical gap in regional antimicrobial resistance (AMR) research. Dairy farming is integral to global food production, yet the wastewater generated by these operations is a significant source of environmental and public health concerns, particularly in the context of antibiotic resistance. This study aimed to isolate and identify antibiotic-resistant bacteria from dairy farm wastewater and evaluate their antibiogram profiles to inform effective management strategies. A total of 60 wastewater samples were collected and subjected to conventional bacterial characterization, followed by molecular detection via PCR and 16S rRNA gene sequencing. The study identified Pseudomonas aeruginosa (35%), Escherichia coli (30%), Bacillus subtilis (16.67%), and Acinetobacter junii (8.33%) as the predominant bacterial species. Sequencing results demonstrated high compatibility with reference sequences, confirming the identities of the isolates. Antibiogram analysis revealed significant resistance patterns: P. aeruginosa exhibited the highest resistance to penicillin (85.71%) and amoxicillin (76.19%), while demonstrating greater sensitivity to ciprofloxacin and cotrimoxazole. E. coli showed notable resistance to penicillin (88.89%), amoxicillin, and ceftriaxone, while B. subtilis and A. junii also demonstrated high levels of resistance to multiple antibiotics. Notably, a substantial proportion of the isolates exhibited multidrug resistance (MDR), with MAR indices ranging from 0.37 to 0.75. Moreover, several antibiotic resistance genes (ARGs) including penA, bla (TEM) , bla (CTX-M) , tetA, tetB, tetC, and ermB were detected across the bacterial species, with high prevalence rates in P. aeruginosa and A. junii, suggesting the potential for horizontal gene transfer and further spread of resistance. These findings underscore the critical need for a One Health approach to mitigate the risks posed by antibiotic-resistant bacteria in dairy farm wastewater, emphasizing the critical importance of responsible antibiotic use and sustainable farming practices to protect public health and environmental integrity.202540458482
256280.9997Characterisation of Antimicrobial Resistance in Special-Fed Veal Production Environments. INTRODUCTION: Antimicrobial resistance (AMR) is one of the leading public health threats globally. AMR genes can be transferred between bacteria through lateral gene transfer, and AMR organisms can spread through environments by contaminated water, agriculture and animals. Thus, widespread environmental dissemination of bacteria and lateral gene transfer facilitate AMR transmission pathways. Farm environments in dairy and calf production are known to harbour AMR bacteria that pose a risk for food contamination and to workers in direct or indirect contact with animals. Escherichia coli is present in farm environments and is known to participate in lateral gene transfer, providing a good marker of resistance genes in each environment. METHODS: In this study, E. coli from nine cohorts of calves was isolated at different time points from nine barns, nine trailers and one slaughterhouse environment in a single special-fed veal calf production facility. The antimicrobial susceptibility to 15 antimicrobials, classified as highly or critically important by the World Health Organization, was characterised for E. coli isolates using Kirby-Bauer disk diffusion. RESULTS: The highest proportion of isolates showing multidrug resistance was present in barn environments (51.7%), where calves were housed from their arrival at < 2 weeks of age until they were transported to slaughter. Additionally, 15 E. coli isolates were resistant to 11 of the 15 antimicrobials tested. Trailer and slaughterhouse environments had greater prevalence of resistance after accommodating calves, including resistance to third-generation cephalosporins. CONCLUSION: These data highlight the importance of calf environments in the dissemination of resistant bacteria and gives insight into where interventions could be most effective in combatting antimicrobial-resistant bacteria that could infect humans and livestock.202539402773
196790.9997Identification of molecular determinants of antibiotic resistance in some fish farms of Ghana. Antimicrobial resistance is a global health challenge caused by the ability of microorganisms including bacteria, fungi, protozoans and viruses to survive the effects of drugs that hitherto were effective against them. This study sought to investigate the presence of antibiotic-resistant bacteria and their corresponding molecular determinants in fish farms of the Central and Western Regions of Ghana. Management practices and antibiotic use at the fish farms were obtained through the administration of a questionnaire. Coliform and Gram-positive bacterial loads of catfish (Clarias gariepinus), tilapia (Oreochromis niloticus) intestinal microbiota, and pond water samples recovered on MacConkey Agar and Mannitol Salt Agar were determined. Bacterial isolates were identified using various biochemical assays. Antibiotic resistance profiles and possible responsible genes of bacterial isolates were determined using the disc diffusion and Polymerase Chain Reaction (PCR) methods respectively. The study revealed that none of the fish farm managers admitted using antibiotics for prevention and treatment of diseases and no major disease outbreak had ever been recorded. Bacterial loads of pond water exceeded the acceptable level of ≤100 E. coli and <10 coliforms per mL for wastewater recommended for use in fish farming. In all, 145 bacterial isolates comprising 99 Gram negative and 46 Gram-positive bacteria were stored and identified. Most isolates were resistant to at least an antibiotic. Both Gram-negative and Gram-positive bacteria were highly resistant to beta-lactam antibiotics with a corresponding high percentage detection of the bla (TEM) gene compared to other classes of antibiotics. This study has revealed the presence of various molecular determinants of antibiotic resistance including bla (TEM), cmIA, qnrS, tetB and bla (CTX-M), in multidrug-resistant bacteria at some fish farms in Ghana. There is the need to increase awareness about risks associated with the misuse and overuse of antibiotics by humans and the potential risk of spread of multi-drug resistant-bacteria in the environment.202236097488
2583100.9997From Farm to Slaughter: Tracing Antimicrobial Resistance in a Poultry Short Food Chain. Background: Short food supply chains are commonly perceived as more sustainable and safer alternatives to conventional production systems, often linked to organic, free-range livestock practices. Materials and methods: This study investigates, for the first time, the distribution of antimicrobial resistance genes (ARGs) and characterizes the microbial communities' composition, using 16S rRNA sequencing and real-time PCR, respectively. Eleven fecal, 76 slaughterhouse surface, 11 cecal, and 11 carcass samples, from 11 poultry farms belonging to the same short food chain, were analyzed in the study. Results: While cleaning and disinfection procedures appeared to reduce the bacterial load on slaughterhouse surfaces, diverse and potentially resistant bacteria, including genera such as Staphylococcus and Streptococcus, persisted both before and after slaughter. ARGs conferring resistance to high-priority critically important antimicrobials (HPCIAs), such as fluoroquinolones and third-generation cephalosporins, were frequently detected on carcasses, with qnrS (76.15%, 95%CI 68.02-84.28%) and bla(CMY2) (57.8%, 95%CI 48.38-67.22%) being the most prevalent. The slaughtering process emerged as a critical step for ARG dissemination via intestinal bacteria, such as genus Lactobacillus. Additionally, the detection of mcr genes and bla(NDM) on carcasses but not in the bird gut samples suggests possible anthropogenic contamination. Discussion: These findings highlight that the evisceration process, slaughterhouse environment, and personnel are all contributing factors in ARG spread and underscore the need for enhanced hygiene protocols and reduced gut ARG carriage in domestic birds to mitigate the risk for the consumer.202540558194
2533110.9997Colistin Resistant mcr Genes Prevalence in Livestock Animals (Swine, Bovine, Poultry) from a Multinational Perspective. A Systematic Review. The objective of this review is to collect and present the results of relevant studies on an international level, on the subject of colistin resistance due to mcr genes prevalence in livestock animals. After a literature search, and using PRISMA guidelines principles, a total of 40 swine, 16 bovine and 31 poultry studies were collected concerning mcr-1 gene; five swine, three bovine and three poultry studies referred to mcr-2 gene; eight swine, one bovine, two poultry studies were about mcr-3 gene; six swine, one bovine and one poultry manuscript studied mcr-4 gene; five swine manuscripts studied mcr-5 gene; one swine manuscript was about mcr-6, mcr-7, mcr-8, mcr-9 genes and one poultry study about mcr-10 gene was found. Information about colistin resistance in bacteria derived from animals and animal product foods is still considered limited and that should be continually enhanced; most of the information about clinical isolates are relative to enteropathogens Escherichia coli and Salmonella spp. This review demonstrates the widespread dispersion of mcr genes to livestock animals, indicating the need to further increase measures to control this important threat for public health issue.202134822638
2558120.9997Antimicrobial resistance in wild game mammals: a glimpse into the contamination of wild habitats in a systematic review and meta-analysis. BACKGROUND: Wild game meat has over the years gained popularity across the globe as it is considered a food source with high protein content, low fat content, and a balanced composition of fatty acids and minerals, which are requirements for a healthy diet. Despite this popularity, there is a concern over its safety as many species of wildlife are reservoirs of zoonotic diseases including those of bacterial origin, more so antibiotic-resistant bacteria. METHODS: This study aimed to describe the prevalence of antibiotic-resistant bacteria in mammalian wild game, following the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. RESULTS: The overall pooled prevalence of antibiotic resistance was established at 59.8% while the prevalence of multidrug resistance (MDR) was 17.2%. Resistance was reported in 32 wild game species and the meta-analysis revealed the highest prevalence of antibiotic resistance in Yersinia spp. (95.5%; CI: 76.8 - 100%) followed by Enterococcus spp. (71%; CI: 44.1 - 92%), Salmonella spp. (69.9%; CI: 44.3 - 90.0%), Staphylococcus spp. (69.3%; CI: 40.3 - 92.3%), and Escherichia coli (39.5%; CI: 23.9 - 56.4%). Most notably, resistance to highest priority, critically important antimicrobials, was recorded in all genera of bacteria studied. Additionally, a significantly higher prevalence of antibiotic resistance was observed in studies conducted in remote settings than those in the vicinity of anthropogenic activities, pointing to extensive contamination of wild habitats. CONCLUSION: This review shows the presence of antibiotic resistance and the carriage of antimicrobial resistance (AMR) genes by bacteria isolated from mammalian wild game species. This is a cause for concern if critical steps to prevent transmission to humans from meat and meat products are not applied in the wild game meat production chain. The extensive occurrence of antibiotic resistance in the wild calls for expansion and adaptation of future AMR surveillance plans to include areas with various anthropogenic pressures including in sylvatic habitats.202539799360
2561130.9997Longitudinal assessment of antibiotic resistance gene profiles in gut microbiomes of infants at risk of eczema. BACKGROUND: While there is increasing knowledge about the gut microbiome, the factors influencing and the significance of the gut resistome are still not well understood. Infant gut commensals risk transferring multidrug-resistant antibiotic resistance genes (ARGs) to pathogenic bacteria. The rapid spread of multidrug-resistant pathogenic bacteria is a worldwide public health concern. Better understanding of the naïve infant gut resistome may build the evidence base for antimicrobial stewardship in both humans and in the food industry. Given the high carriage rate of extended spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in Asia, we aimed to evaluate community prevalence, dynamics, and longitudinal changes in antibiotic resistance gene (ARG) profiles and prevalence of ESBL-producing E. coli and K. pneumoniae in the intestinal microbiome of infants participating in the Growing Up in Singapore Towards Healthy Outcomes (GUSTO) study, a longitudinal cohort study of pregnant women and their infants. METHODS: We analysed ARGs in the first year of life among 75 infants at risk of eczema who had stool samples collected at multiple timepoints using metagenomics. RESULTS: The mean number of ARGs per infant increased with age. The most common ARGs identified confer resistance to aminoglycoside, beta-lactam, macrolide and tetracycline antibiotics; all infants harboured these antibiotic resistance genes at some point in the first year of life. Few ARGs persisted throughout the first year of life. Beta-lactam resistant Escherichia coli and Klebsiella pneumoniae were detected in 4 (5.3%) and 32 (42.7%) of subjects respectively. CONCLUSION: In this longitudinal cohort study of infants living in a region with high endemic antibacterial resistance, we demonstrate that majority of the infants harboured several antibiotic resistance genes in their gut and showed that the infant gut resistome is diverse and dynamic over the first year of life.202032345218
5610140.9997Characterization of antimicrobial resistance profiles in Escherichia coli isolated from captive mammals in Ecuador. BACKGROUND: This study focuses on the AMR profiles in E. coli isolated from captive mammals at EcoZoo San Martín, Baños de Agua Santa, Ecuador, highlighting the role of wildlife as reservoirs of resistant bacteria. AIMS: The aim of this research is to investigate the antimicrobial resistance profiles of E. coli strains isolated from various species of captive mammals, emphasizing the potential zoonotic risks and the necessity for integrated AMR management strategies. MATERIALS & METHODS: A total of 189 fecal samples were collected from 70 mammals across 27 species. These samples were screened for E. coli, resulting in 90 identified strains. The resistance profiles of these strains to 16 antibiotics, including 10 β-lactams and 6 non-β-lactams, were determined using the disk diffusion method. Additionally, the presence of Extended-Spectrum Beta-Lactamase (ESBL) genes and other resistance genes was analyzed using PCR. RESULTS: Significant resistance was observed, with 52.22% of isolates resistant to ampicillin, 42.22% to ceftriaxone and cefuroxime, and 27.78% identified as ESBL-producing E. coli. Multiresistance (resistance to more than three antibiotic groups) was found in 35.56% of isolates. Carnivorous and omnivorous animals, particularly those with prior antibiotic treatments, were more likely to harbor resistant strains. DISCUSSION: These findings underscore the role of captive mammals as indicators of environmental AMR. The high prevalence of resistant E. coli in these animals suggests that zoos could be significant reservoirs for the spread of antibiotic-resistant bacteria. The results align with other studies showing that diet and antibiotic treatment history influence resistance profiles. CONCLUSION: The study highlights the need for an integrated approach involving veterinary care, habitat management, and public awareness to prevent captive wildlife from becoming reservoirs of antibiotic-resistant bacteria. Improved waste management practices and responsible antibiotic use are crucial to mitigate the risks of AMR in zoo environments and reduce zoonotic threats.202439016692
1926150.9997Whole genome sequencing revealed high occurrence of antimicrobial resistance genes in bacteria isolated from poultry manure. BACKGROUND: Global demand for food has driven expansion and intensification of livestock production, particularly in developing nations where antibiotic use is often routine. Waste from poultry production, including manure, is commonly utilized as fertilizers in agroecosystems, risking environmental contamination with potentially zoonotic bacteria and antimicrobial resistance genes (ARGs). METHODS: Here, 33 bacterial isolates were recovered from broiler (n = 17) and layer (n = 16) chicken manure by aerobic culture using Luria Bertani agar. Antimicrobial susceptibility testing (AST) was performed using disc diffusion method. MALDI-ToF and 16S rRNA sequencing were used to identify and compare a subset of antibiotic-resistant isolates (n = 13). Comparison of whole genome sequence assemblies and phenotypic assays were used to assess capacity for biofilm formation, heavy metal tolerance and virulence. RESULTS: AST by disc diffusion revealed all isolates were resistant to a minimum of three antibiotics, with resistance to ampicillin, co-trimoxazole, fluoroquinolones, tetracyclines, streptomycin, rifampicin and/or chloramphenicol detected. Stutzerimonas sp. and Acinetobacter sp. were the common genera observed in this study. Genome sequencing of each selected isolate revealed carriage of multiple ARGs capable of conferring resistance to many antimicrobials commonly employed in poultry production and human medicine, including tetracyclines, quinolones, macrolides, sulfonamide and cephalosporins. CONCLUSIONS: The high occurrence of ARGs in studied bacterial isolates confirms that poultry manure could act as a source of genetic material that could be transferred to commensal microbiota and opportunistic pathogens of humans. Understanding the complex resistome interplay between humans, animals, and the environment requires a One Health approach, with implications for agricultural settings and public health.202539880102
5667160.9997Quantifying antibiotic impact on within-patient dynamics of extended-spectrum beta-lactamase resistance. Antibiotic-induced perturbation of the human gut flora is expected to play an important role in mediating the relationship between antibiotic use and the population prevalence of antibiotic resistance in bacteria, but little is known about how antibiotics affect within-host resistance dynamics. Here we develop a data-driven model of the within-host dynamics of extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae. We use bla(CTX-M) (the most widespread ESBL gene family) and 16S rRNA (a proxy for bacterial load) abundance data from 833 rectal swabs from 133 ESBL-positive patients followed up in a prospective cohort study in three European hospitals. We find that cefuroxime and ceftriaxone are associated with increased bla(CTX-M) abundance during treatment (21% and 10% daily increase, respectively), while treatment with meropenem, piperacillin-tazobactam, and oral ciprofloxacin is associated with decreased bla(CTX-M) (8% daily decrease for all). The model predicts that typical antibiotic exposures can have substantial long-term effects on bla(CTX-M) carriage duration.202032379042
5665170.9997Complementarity of Selective Culture and qPCR for Colistin Resistance Screening in Fresh and Frozen Pig Cecum Samples. Retrospective studies involving the screening of frozen stored collections of samples are commonplace when a new threat emerges, but it has been demonstrated that the freeze-thaw process can affect bacterial viability. The study of colistin-resistant bacteria in human and animal samples is an example of this issue. In this study, we compared culture-based and PCR-based methods for analyzing relative occurrence and diversity of colistin-resistant bacteria in caecal samples to determine the most appropriate method for frozen samples. Thus, 272 samples from the caecal contents of healthy pigs were tested before and after a 6-month freezing period. A selective medium was used when traditional isolation of colistin-resistant bacteria was tested, while a real-time SYBR(®) Green I PCR assay was applied for mcr-1 quantification. The number of samples with colistin-resistant isolates was higher in fresh samples (247/272) than in frozen ones (67/272) and showed a higher diversity of colistin-resistant genera. PCR identification of mcr colistin resistance genes evidenced that mcr-1 was the most prevalent mcr gene and mcr-2 was detected for the first time in pigs from Spanish animal production. The number of samples with mcr-1-carrying bacteria after a freezing period decreased, while real-time quantitation of the mcr-1 gene showed similar values in frozen and fresh samples. Therefore, when frozen cecal samples need to be analyzed, molecular detection of DNA could be the best option to provide a highly representative frame of the initial population present in the sample, and culture-based methods might be a useful complement to study colistin resistance levels.202033240230
2843180.9997High Throughput Screening of Antimicrobial Resistance Genes in Gram-Negative Seafood Bacteria. From a global view of antimicrobial resistance over different sectors, seafood and the marine environment are often considered as potential reservoirs of antimicrobial resistance genes (ARGs) and mobile genetic elements (MGEs); however, there are few studies and sparse results on this sector. This study aims to provide new data and insights regarding the content of resistance markers in various seafood samples and sources, and therefore the potential exposure to humans in a global One Health approach. An innovative high throughput qPCR screening was developed and validated in order to simultaneously investigate the presence of 41 ARGs and 33 MGEs including plasmid replicons, integrons, and insertion sequences in Gram-negative bacteria. Analysis of 268 seafood isolates from the bacterial microflora of cod (n = 24), shellfish (n = 66), flat fishes (n = 53), shrimp (n = 10), and horse mackerel (n = 115) show the occurrence of sul-1, ant(3″)-Ia, aph(3')-Ia, strA, strB, dfrA1, qnrA, and bla(CTX-M-9) genes in Pseudomonas spp., Providencia spp., Klebsiella spp., Proteus spp., and Shewanella spp. isolates and the presence of MGEs in all bacterial species investigated. We found that the occurrence of MGE may be associated with the seafood type and the environmental, farming, and harvest conditions. Moreover, even if MGE were detected in half of the seafood isolates investigated, association with ARG was only identified for twelve isolates. The results corroborate the hypothesis that the incidence of antimicrobial-resistant bacteria (ARB) and ARG decreases with increasing distance from potential sources of fecal contamination. This unique and original high throughput micro-array designed for the screening of ARG and MGE in Gram-negative bacteria could be easily implementable for monitoring antimicrobial resistance gene markers in diverse contexts.202235744743
1925190.9997Camel Milk Resistome in Kuwait: Genotypic and Phenotypic Characterization. Antimicrobial resistance (AMR) is one of the major global health and economic threats. There is growing concern about the emergence of AMR in food and the possibility of transmission of microorganisms possessing antibiotic resistance genes (ARGs) to the human gut microbiome. Shotgun sequencing and in vitro antimicrobial susceptibility testing were used in this study to provide a detailed characterization of the antibiotic resistance profile of bacteria and their ARGs in dromedary camel milk. Eight pooled camel milk samples, representative of multiple camels distributed in the Kuwait desert, were collected from retail stores and analyzed. The genotypic analysis showed the presence of ARGs that mediate resistance to 18 classes of antibiotics in camel milk, with the highest resistance to fluoroquinolones (12.48%) and disinfecting agents and antiseptics (9%). Furthermore, the results pointed out the possible transmission of the ARGs to other bacteria through mobile genetic elements. The in vitro antimicrobial susceptibility testing indicated that 80% of the isolates were resistant to different classes of antibiotics, with the highest resistance observed against three antibiotic classes: penicillin, tetracyclines, and carbapenems. Multidrug-resistant pathogens including Klebsiella pneumoniae, Escherichia coli, and Enterobacter hormaechei were also revealed. These findings emphasize the human health risks related to the handling and consumption of raw camel milk and highlight the necessity of improving the hygienic practices of farms and retail stores to control the prevalence of ARGs and their transmission.202438786109