Background Our program was initiated to manage an unexpected increase in carbapenem-resistant Enterobacterales (CRE) contamination identified during routine environmental surveillance in the small animal intensive care unit (ICU) at The Ohio State University Veterinary Medical Center (VMC). Addressing CRE, an urgent public health threat, was critical to protect patients and veterinary personnel.Hypothesis/Objectives Design an active surveillance program to guide infection control and prevention (ICP) measures and successfully manage CRE in the VMC environment.Methods The VMC's environmental surveillance program has conducted monthly surveillance since 2018 to identify pathogens, including CRE. In response to increased CRE contamination in September 2023, a CRE Response Plan Working Group developed and implemented a plan to control contamination and prevent disease transmission. Interventions included enhanced surveillance, strengthened ICP measures, updated CRE protocols, improved cleaning and disinfection, hand hygiene audits and education, ICP training, and a unified communication strategy.Results Weekly environmental surveillance identified 4 repeatedly CRE-contaminated surfaces: the covered outdoor run, the emergency room and ICU hallway floors, and the ICU medication preparation counter and CUBEX machine. One patient with a clinical CRE infection was managed without any further identified transmission. By January 2024, repeated CRE contamination was no longer detected in the VMC. Screening ICU patients for CRE fecal colonization identified a 1.9% prevalence.Conclusions and clinical importance Active environmental surveillance enabled early detection of increased CRE contamination which guided ICP measures, decreasing the risk of CRE transmission. We provide a framework for responding to nosocomial threats in veterinary hospital settings.

Enterobacterales that are resistant to carbapenem (CRE) pose a major threat to public health. There is limited data on the epidemiology of CRE genotypic alleles circulating in the natural environment, particularly in Africa. This paper describes the epidemiology of CRE strains with specific plasmid replicons, sequence types, and resistance determinants, as identified in isolates from Nairobi River surface water, a wastewater treatment plant (WWTP), and slaughterhouse discharges in Nairobi, Kenya. From 336 total CRE isolates, 150 (44.6%) were selected for whole-genome sequencing. Six different genotypic alleles such as blaKPC-2, blaNDM-7, blaNDM-5, blaOXA-181, blaNDM-1, and blaOXA-232 were found. The blaNDM gene alleles, predominantly blaNDM-5 (n = 118, 78.6% of all alleles), were commonly detected in Escherichia coli. blaNDM-1 (n = 10, 6.6%) was detected in all Enterobacter spp. from WWTP, and Nairobi River, and blaNDM-7 (n = 7, 4.6%) in K. pneumoniae from WWTP, E. coli, and E. hormaechei from Nairobi River. The blaOXA variants were rare (blaOXA-181 n = 8, 5.3%; blaOXA-232 n = 3, 2%) while the least detected was blaKPC-2 (n = 5, 3.3%) in Enterobacter species from WWTP. The blaOXA-181 and blaOXA-232 variants were present in isolates of K. pneumoniae, E. coli, and E. hormaechei that co-harbored either blaNDM-1 or blaNDM-5 from WWTP and the Nairobi River. E. cloacae was detected carrying blaOXA-181, while K. pneumoniae isolates harboured blaOXA-181 and blaOXA-232, respectively, from WWTP. K. pneumoniae harboring blaNDM-5 co-harboring blaOXA-181 or blaOXA-232 was also identified. The most common sequence type (ST) found by multilocus sequence types (MLST) analysis was E. coli ST167 carrying blaNDM-5 (n = 29). K. pneumoniae ST147 carrying blaNDM-5 was also found in isolates co-harboring blaOXA-181 and blaOXA-232 variants. It's interesting to note that some Enterobacter species ST413 harboured blaNDM-1, while others harboured blaKPC-2, and one Citrobacter freundii ST964 was detected carrying blaNDM-5 of all CRE isolates. The presence of diverse CRE genotypes in surface water, wastewater, and slaughterhouse effluent in Nairobi County, Kenya, poses a public health threat that requires urgent and targeted intervention strategies.

Antibiotic-resistant infections cause an estimated 2.8 million illnesses and 35,900 deaths annually in the USA. Carbapenems are a class of antibiotics that are generally reserved to treat life-threatening invasive infections including sepsis. Accurate diagnosis of carbapenem-resistant infections is critical for early and appropriate treatment. bla IMP encodes bacterial production of the IMP metallo-beta-lactamase (MBL), which can confer resistance to all the beta-lactams including carbapenems. Zinc is an essential co-factor in the IMP MBL enzymatic hydrolysis of carbapenems. Tests for the presence of IMP carbapenemase, such as the Carba NP, include zinc sulphate (ZnSO 4 ) although broth dilution methods for determining MIC for carbapenems may vary. We hypothesized that ZnSO 4 availability would improve the accuracy of carbapenem MIC determination for bacteria expressing bla IMP . Thus, the objective of this study was to determine if supplemental ZnSO 4 affects the carbapenem MICs of Enterobacterales , Alteromonadales and Moraxellales expressing bla IMP . Isolates utilized for this study were originally recovered from environmental samples collected at farms, wastewater treatment plants and surface water. They were selected based on phenotypic non-susceptibility to carbapenems and genetic confirmation of bacterial carriage of bla IMP . Cation-adjusted Mueller–Hinton broth suspensions of each isolate standardized to a 0.5 MacFarland standard were tested with and without ZnSO 4 added at 0.1 mmol l −1 concentration to determine MICs using standard extended-spectrum beta-lactamase microbroth dilution MIC panels. Although we observed that Morganellaceae imipenem MICs were higher ( P <0.001) than those from other bacteria harbouring bla IMP , the inclusion of supplemental ZnSO 4 did not influence carbapenem MIC. This suggests that supplemental ZnSO 4 will not improve the accuracy of carbapenem MICs in environmental bacteria expressing IMP carbapenemase. Additional research will be required to identify important factors that may influence the expression of carbapenemase including IMP and the accurate determination of clinical MICs, which is critical to appropriate therapeutic decision-making.

Objective:To identify host-specific pathogenesis and intraspecies transmission potential of spatiotemporally related Escherichia coli causing human and canine urinary tract infections (UTIs). Methods:We collected 103 unique E coli isolates that caused clinical UTIs from a veterinary and human teaching facility over a 2-year period. Isolates were assessed for antimicrobial resistance and biofilm production and sequenced to compare population structure, functional pathways, and virulence and resistance gene composition. Results:Humans' E coli were more likely to be from sequence type (ST)-131 and ST95, whereas canine isolates were more likely to be from ST372. Sequence type 73, ST38, and ST12 were composed of equal proportions of human and canine isolates. Escherichia coli from both humans and canines were significantly enriched in different functional pathways that promote colonization of the urinary tract. Canine isolates carried a more robust virulome (β, 14.3; 95% CI, 5.8 to 22.9) that was enriched in many virulence genes and had greater biofilm production (OR, 2.91; 95% CI, 1.24 to 6.81). Human isolates carried a more abundant resistome (Incidence Risk Ratio [IRR], 1.69; 95% CI, 1.31 to 2.18) that conferred reduced susceptibility to multiple drug classes. Conclusions:Urinary-associated E coli from different hosts are composed of unique strain populations that harbor different functional, virulence, and resistance capacities. Escherichia coli from ST73, ST38, and ST12 can cause infections in both species, suggesting potential for cross-species transfer. Clinical Relevance:Our findings contribute to a broader understanding of E coli UTIs in dogs, how they differ from human infections, and the risk for cross-species transmission of specific E coli subgroups.

Carbapenemase-producing Enterobacterales (CPE) have emerged as an important nosocomial threat to hospitalized patients, but CPE can also colonize the enteric microbiota of healthy individuals in the community. We hypothesized that clinically relevant CPE are frequently transported in municipal wastewater flows to treatment plants where they are reduced but not eliminated and are subsequently discharged into nearby surface waters and disseminate in the environment. We sampled untreated influent, treated effluent, and nearby surface waters weekly for a one-year period at a single large metropolitan wastewater treatment plant (WWTP) servicing Columbus, Ohio USA. In addition, we investigated the dissemination of these CPE into the environment and the downstream watershed, including sediment, fish, mammals, waterfowl, crops and soils. CPE were recovered from each (100 %) of the 44 influent samples and from 37 (84 %) effluent samples. We also isolated CPE from 50 % (22/44) of the upstream water, 82 % (36/44) of downstream, and 68 % (30/44) way downstream water. CPE were most commonly Enterobacter spp. expressing the blaKPC-2 genotype, although a variety of other species and genotypes were observed. blaKPC concentration was greatest in the influent (mean = 106 gene copies/100 mL water) and treatment resulted in a 4-log reduction in blaKPC concentration (P < 0.05), which was consistent with reduction in total bacteria concentration. We found 22 of 450 fish vent swabs (4.9 %) carrying CPE, but CPE were not recovered from terrestrial wildlife living in the Scioto watershed. Fish intestinal microbiome maintained approximately 1.5 × 104 copies of blaKPC per gram of feces. Our data support the hypothesized flow of CPE from healthcare settings into surface water and the downstream natural environment via municipal wastewater discharge. Our results suggest that river environments can serve as a reservoir for CPE, facilitating their broader One Health dissemination among surface water, wildlife, agriculture, and ultimately back to humans.

The blaIMP resistance gene encodes a metallo-beta-lactamase in bacteria, which confers reduced susceptibility or resistance to all the beta-lactams, including carbapenems which are critical for treating life-threatening infections. The dissemination of blaIMP among various taxonomic families shows the diversity and range of horizontal gene transfer. Using short-read whole genome sequencing and bioinformatic tools, we determined the genetic motifs surrounding blaIMP present in 32 bacterial isolates recovered from environmental sources and agriculture facilities. blaIMP can be located extra-chromosomally on plasmids or within incomplete and complete Tn7 chromosomal structures. We identified a complete Tn7 transposon harboring the blaIMP-27 gene cassette within a class 2 integron located in chromosomal contigs of Shewanella spp. and Providencia spp. Acinetobacter spp. isolates were observed with truncated and incomplete Tn7 transposons, while conserving the class 2 integron and resistance gene cassettes. Additionally, IncQ1 plasmids carried by Proteus spp., Escherichia coli, and other Enterobacteriaceae spp. harbored class 2 integrons with blaIMP-64 and sat2 resistance gene cassettes. In an Acidovorax sp. isolate, blaIMP-27 and sat2 gene cassettes were found associated with an insertion sequence, ISL3 transposase, in an RP4 plasmid. The conserved structure of Tn7 in Shewanella spp. and Providencia spp. is consistent with these species being potential reservoirs from which other bacterial species have acquired partial Tn7 motifs, and the blaIMP-27 gene cassette. These data contribute to a broader understanding of the dissemination and temporality of blaIMP alleles and their mobile genetic elements.

Antimicrobial-resistant bacteria, including both pathogens and commensal organisms, can be introduced into livestock populations by a variety of mechanisms including contaminated water, animal and human movement, and animal feed. We hypothesized that animal feed is an important mechanism for introduction of both Salmonella and commensal reservoir bacteria harboring mobile antimicrobial-resistance genes into livestock populations. The identification of high-risk feed components may allow targeted interventions that will reduce carriage of these organisms in food animals and ultimately improve food safety. We collected 666 livestock feed component samples from 16 individual local mills (n = 263) and from 2 corporations with multiple feed mills located across the US (n = 403). Salmonella enterica were recovered from 7.2 % of samples (n = 48) and were more likely to be found (P < 0.02) in blended feeds, animal protein components, and oilseed meals. We also recovered two carbapenem-resistant Enterobacterales isolates (0.3 %), both E. coli sequence type 167 O101:H9 carrying an IncF plasmid mediated blaNDM-5, from two dried distiller's grain samples sourced from two states in the upper Midwest. Antimicrobial-resistant commensal bacteria were found in all feed component types, although the overall prevalence of each resistance phenotype was below seven percent except for AmpC β-lactamase producing strains (19.5 %). The identification of feed components with higher risk of contamination suggests that targeted interventions could potentially reduce feed contamination. Contamination reduction efforts focusing on feed components with heating and cooling steps may reduce the proliferation of pathogens and resistant bacterial strains in finished feed fed to livestock.

Zoonotic pathogens, including Salmonella and antimicrobial resistant bacteria, may contaminate the food or treats consumed by our pets. These may directly impact the health of the pets or may be transferred to humans who are in close contact. To better understand the potential risk, we purchased 505 pet treats from pet and farm supply stores, grocery stores, and online retailers in the U.S. over a period of 16 months to identify and characterize Salmonella and Enterobacterales resistant to Highest Priority Critically Important Antimicrobials. We used selective media to detect Salmonella and bacteria resistant to colistin, carbapenems, fluoroquinolones, and 3rd and 4th generation cephalosporins. Four pig ear treats from Brazil were positive for Salmonella, with serotypes, Muenchen, Derby, Agona and Regent. We found that S. Muenchen and S. Derby were closely related to clinical and environmental isolates from the U.S., Canada, Venezuela, and Colombia. We detected three colistin resistant isolates, Klebsiella pneumoniae, Escherichia coli, and Enterobacter hormaechei, all from pig ear treats from Brazil, harboring the mcr-1.18 resistance gene on identical IncX4 plasmids. In addition, we recovered one carbapenem resistant E. coli harboring both blaKPC-2 and blaNDM-5 from a "bully stick". We found that treats originating from North America and treats purchased in grocery stores had a lower risk of contamination with bacteria resistant to the antimicrobials tested. Outreach and extension activities are needed to increase awareness of the risks of contaminated pet treats and to highlight the importance of hand hygiene when feeding and interacting with pets.

2. Abstract Antibiotic-resistant infections cause an estimated 2.8 million illnesses and 35,900 deaths annually in the US. Carbapenems are a class of antibiotics that are generally reserved to treat life-threatening invasive infections including sepsis. Accurate diagnosis of carbapenem-resistant infections is critical for early and appropriate treatment. blaIMP encodes bacterial production of the IMP metallo-beta-lactamase (MBL) which can confer resistance to all the beta-lactams including carbapenems. Zinc is an essential co-factor in the IMP MBL enzymatic hydrolysis of carbapenems. Tests for the presence of IMP carbapenemase, such as the Carba NP, include zinc-sulfate (ZnSO4) although broth dilution methods for determining minimum inhibitory concentration (MIC) for carbapenems do not. We hypothesized that ZnSO4 availability would improve the accuracy of carbapenem MIC determination for Enterobacterales expressing blaIMP. Thus, the objective of this study was to determine if supplemental ZnSO4 affects the carbapenem MICs of Enterobacterales and other bacteria expressing blaIMP. Isolates utilized for this study were originally recovered from environmental samples collected at farms, wastewater treatment plants, and from surface water. They were selected based on phenotypic non-susceptibility to carbapenems and genetic confirmation of bacterial carriage of blaIMP. Cation-adjusted Mueller-Hinton broth suspensions of each isolate standardized to a 0.5 MacFarland standard were tested with and without ZnSO4 at 0.1 mmol/L concentration to determine MICs using standard extended-spectrum beta-lactamase micro-broth dilution MIC panels. Although we observed that Morganellaceae imipenem MICs were higher (P < 0.001) than those from other Enterobacterales harboring blaIMP, the inclusion of supplemental ZnSO4 did not influence carbapenem MIC. Additional research will be required to identify important factors that may influence the expression of carbapenemase including IMP, and the accurate determination of clinical MICs which is critical to appropriate therapeutic decision-making.

Background: In 2018, the Ohio State University College of Veterinary Medicine (OSU CVM) implemented an Antimicrobial Stewardship Program, central to which was the integration of an environmental surveillance (ES) program. The ES focuses on pathogens recognized as urgent threats to public health by the Centers for Disease Control and Prevention. The pathogens currently targeted include carbapenemase-producing Enterobacterales (CPE), Salmonella spp., methicillin-resistant Staphylococcus spp. (MRSs), vancomycin resistant Enterococcus spp., and enrofloxacin resistant Pseudomonas aeruginosa. Identification of these pathogens allows the hospital to be aware of the local environmental microflora which can act as a sentinel for disease in the hospital, potentially causing healthcare associated infections. Therefore, the objective of this program is to identify resistant bacterial pathogens, characterize their resistance profiles, analyze prevalence patterns, and initiate infection control interventions where needed in the OSU VMC. Method: From January 2018 through December 2023, a total of 5449 samples were collected from approximately 86 locations across the OSU VMC encompassing the small animal, equine, and farm animal sections. A majority (64%, n=3561) of samples were collected from the small animal hospital, with the farm animal section contributing 1055 samples and the equine section 899. Areas sampled were frequented by both humans and animals, as well as surfaces exclusively touched by humans. Samples were collected using Swiffers® and processed through selective culture media. Result: Approximately half (52%, n=2890) of the samples collected represented human-touch only surfaces. A total of 3794 bacterial isolates were recovered, with an overall low prevalence for all targeted pathogens. Prevalence of CPE was 2% (n=103), with Enterobacter species being the most common. Recovery of MRSs was 8.5% (n=464) and Salmonella species was 1% (n=47). Conclusion: Through this initiative, the equine division of the OSU VMC collaborated with the antimicrobial stewardship team to enhance their Salmonella fecal and ES practices. In 2019, ES was critical in identifying persistent CPE and extended-spectrum cephalosporin-resistant Enterobacteriaceae in the ICU and surrounding areas of the small animal hospital. Effective measures were taken to halt the spread of ESC among patients and eliminate CPE in the environment. With the discovery of a new CPE in early 2023 in the small animal ICU and nearby areas, the program initiated targeted ES and cleaning and disinfection protocols, to identify contaminated areas and control disease transmission. These efforts have increased patient safety, health, and well-being, demonstrating how ES can be an important tool for infection control and prevention in veterinary settings.

The global food trade provides a means of disseminating antimicrobial resistant (AMR) bacteria and genes. Using selective media, carbapenem-resistant species of Enterobacterales (Providencia sp. and Citrobacter sp.), were detected in a single package of imported frozen shrimp purchased from a grocery store in Ohio, USA. Polymerase chain reaction confirmed that both isolates harbored bla(NDM-1) genes. Following PacBio long read sequencing, the sequences were annotated using the NCBI Prokaryotic Genome Annotation Pipeline. The bla(NDM-1) genes were found in IncC plasmids, each with different antimicrobial resistance island configuration. We found that the bla(NDM-1) AMR islands had close relationships with previously reported environmental, food, and clinical isolates detected in Asia and the United States, highlighting the importance of the food chain in the global dissemination of antimicrobial resistance.

The dissemination of antibiotic resistance (AR) through various environments and the role of AR hotspots in public health crises are gaining increasing attention. Aquatic biofilms are speculated to play a significant role in AR spread due to their collection of diverse microorganisms and facilitation of horizontal gene transfer (HGT). However, few studies have characterized the AR genes (resistome) present in natural river biofilms. The goal of this study was to use MinION long-read sequencing to analyze the microbiome, resistome, and mobile genetic elements (MGEs) in periphyton (epilithic biofilms) (n = 56) from a multiuse watershed in Ohio, to elucidate the role of periphyton in clinically relevant AR. Key members of the periphyton microbiome included Flavobacterium and Aeromonas. Overall, periphyton microbial communities shifted with season and location. Specifically, species of Porphyrobacter and Cyanobacteria were more abundant in biofilms during the summer season. Potentially pathogenic bacteria, including the family Enterobacteriaceae, the fish pathogen Pseudomonas koreensis, and the human pathogen Shigella flexneri, were more abundant in sites downstream of the large city, Columbus, OH, than upstream. The periphyton resistome carried diverse AR genes for a variety of classes, but had minimal clinical relevance. Escherichia, Escherichia coli, and Muvirus were common hosts of AR genes (ARGs) and MGEs. Pseudomonas and Cyanobacteria were frequently MGE hosts, but not AR genes, indicating the potentially important role of these taxa in HGT within and around biofilms. While the sequencing depth in this study was relatively shallow, these findings highlight the mobility potential for the transmission of ARGs in river biofilms.

Carbapenem-resistant bacteria (CRB) present a significant global public health concern. Sub-Saharan Africa has borne a heavy burden of CRB with a reported prevalence of up to 60% in some patient populations. es in Africa focus on clinical CRB isolates, with limited data on their spread in the natural environment. Therefore, the purpose of this study was to report the recovery of CRB from Nairobi River surface waters and nearby anthropogenic and zoonotic sources in Nairobi County, Kenya. A total of 336 CRB were recovered from 336 (250 mL) samples, with 230 of the samples (68.5%) producing one or more CRB isolates. CRB were recovered most commonly from untreated sewage influent (100% of 36 samples; 79 total isolates), treated effluent (93% of 118 samples; 116 total isolates), Nairobi River surface waters upstream (100% of 36 samples; 57 total isolates), downstream (100% of 36 samples; 45 total isolates), and way downstream from the wastewater treatment plant (73% of 11 samples; 19 total isolates), slaughterhouse effluent discharges 1.5%, (5/336), animal contact areas 0.9%, (3/336), a manhole sewer from the affluent neighborhood of Karen at 2.7%, (9/336) respectively. The CRB included Escherichia coli (158, 47%), Klebsiella pneumoniae (74, 22%), and Enterobacter spp (43, 13%). Aeromonas spp (29, 9%) Acinetobacter baumannii (12, 3.6%), Citrobacter freundii (7, 2.1%), Pseudomonas aeruginosa (5, 1.5%) and other species (8, 2.4%). CRB genotypes included blaNDM (246, 73.2%), blaKPC (40, 12%), blaVIM (51, 15.2%), blaOXA-48-like (65, 19.3%), blaIMP (15, 4.5%), and blaGES (7, 2.1%). Sixty-nine of the CRB isolates (20.5%) harbored multiple carbapenemase-encoding genes. Our results indicate that clinically important CRB are commonly present in Nairobi River surface water and from nearby wastewater and livestock sources. These pose an important public health threat that requires urgent intervention strategies and additional investigation.

Once considered to be a simple cause-and-effect relationship with localized impact, the concept of how antimicrobial use drives antimicrobial resistance is now recognized as a complex, transdisciplinary problem on a global scale. While the issue of antimicrobial resistance is often studied and addressed at the antimicrobial-human or antimicrobial-animal treatment interface, the role of the environment in the One Health dynamics of antimicrobial resistance is not as well understood. Antimicrobial-resistant bacteria, including those resistant to carbapenem drugs, are emerging in veterinary clinical environments, on farms, and in natural habitats. These multidrug-resistant bacteria can colonize our livestock and companion animals and are later disseminated into the environment, where they contaminate surface waters and colonize wildlife. From here, the One Health transmission cycle of antimicrobial-resistant bacteria is completed as environmental reservoirs can serve as sources of antimicrobial resistance transmission into human or animal healthcare settings. In this review, we utilize a One Health perspective to evaluate how environments become contaminated and, in turn, become reservoirs that can colonize and infect our veterinary species, and how the veterinary field is combating environmental contamination with antimicrobial stewardship regulations and program implementation. The companion Currents in One Health by Parker et al, AJVR , April 2024, addresses the intensive research that justifies this One Health cycle of antimicrobial resistance transmission and emerging techniques that are dissecting the complex interactions at the One Health interface.

Since their commercialization, scientists have known that antimicrobial use kills or inhibits susceptible bacteria while allowing resistant bacteria to survive and expand. Today there is widespread antimicrobial resistance (AMR), even to antimicrobials of last resort such as the carbapenems, which are reserved for use in life-threatening infections. It is often convenient to assign responsibility for this global health crisis to the users and prescribers of antimicrobials. However, we know that animals never treated with antimicrobials carry clinically relevant AMR bacteria and genes. The causal pathway from bacterial susceptibility to resistance is not simple, and dissemination is cyclical rather than linear. Amplification of AMR occurs in healthcare environments and on farms where frequent exposure to antimicrobials selects for resistant bacterial populations. The recipients of antimicrobial therapy release antimicrobial residues, resistant bacteria, and resistance genes in waste products. These are reduced but not removed during wastewater and manure treatment and enter surface waters, soils, recreational parks, wildlife, and fields where animals graze and crops are grown for human and animal consumption. The cycle is complete when a patient carrying AMR bacteria is treated with antimicrobials that amplify the resistant bacterial populations. Reducing the development and spread of AMR requires a One Health approach with the combined commitment of governments, medical and veterinary professionals, agricultural industries, food and feed processors, and environmental scientists. In this review and in the companion Currents in One Health by Ballash et al, JAVMA, April 2024, we highlight just a few of the steps of the complex cyclical causal pathway that leads to the amplification, dissemination, and maintenance of AMR.

1. Public aquariums are organizations that offer their visitors an opportunity to engage with, gain knowledge of, and contribute to the conservation of species and ecosystems. Unfortunately, the ways many aquariums maintain their populations include primarily sourcing marine species from the wild. This creates the potential for negative impacts to stressed aquatic environments and a need for a holistic approach to quantify the impacts of aquarium population management. 2. The objective of this study was to implement a developed One Welfare-Based Ecosystem Model to predict impacts of aquarium population management on humans, animals, and the environment. Ecosystem indicators were selected and grouped into physical-chemical, biological, and socio-economic categories. Data included measurements of environmental and animal indicators at field sites frequently visited by aquariums to collect animals from, aquarium visitor surveys, and publicly available data. Model development included comparing field data to reference information and then adding weightings based on indicator significance and alpha-testing of the model. 3. At model completion, each institution was allocated a One Welfare status. Confidence ratings of measured indicators and reference data occurred for each field site to determine the appropriateness of use of the model for each aquarium. 4. Findings included a positive One Welfare status, with collaborating institutions scoring in the good and excellent categories (81.92/100 and 92.51/100). Confidence ratings showed positive results (62/100 and 78/100) for model applicability for the field sites studied as well as future use by other institutions assessing similar parameters. 5. To maintain their recognition as leaders in conservation, public aquariums must objectively assess their direct and indirect influence on marine ecosystems moving forward.

Wastewater treatment plants (WWTPs) are thought to be a major disseminating source of antibiotic resistance (AR) to the environment, establishing a crucial connection between human and environmental resistome. The objectives of this study were to determine how wastewater effluents impact microbiome and resistome of freshwater and fish, and identify potential AR-carrying clinically relevant pathogens in these matrices. We analyzed wastewater influent and effluent from four WWTPs in three metropolitan areas of Ohio, USA via shotgun metagenomic sequencing. We also sequenced river water and fish guts from three reaches (upstream, at the WWTP outfall, and downstream). Notably, we observed a decline in microbiome diversity and AR gene abundance from wastewater to the receiving river. We also found significant differences by reach and trophic level (diet) in beta-diversity of the fish gut microbiomes. SourceTracker revealed that 0.443 and 0.248 more of the of the fish gut microbiome was sourced from wastewater effluent in fish from the outfall and downstream locations, respectively, compared to upstream fish. Additionally, AR bacteria of public health concern were annotated in effluent and river water samples, indicating potential concern for human exposure. In summary, our findings show the continued role of wastewater as a significant AR reservoir and underscores the considerable impact of wastewater discharge on aquatic wildlife, which highlights the One Health nature of this issue.

Concern about zoonoses and wildlife has increased. Few studies described the role of wild mammals and environments in the epidemiology of Salmonella. Antimicrobial resistance is a growing problem associated with Salmonella that threatens global health, food security, the economy, and development in the 21st century. The aim of this study is to estimate the prevalence and identify antibiotic susceptibility profiles and serotypes of non-typhoidal Salmonella enterica recovered from non-human primate feces, feed offered, and surfaces in wildlife centers in Costa Rica. A total of 180 fecal samples, 133 environmental, and 43 feed samples from 10 wildlife centers were evaluated. We recovered Salmonella from 13.9% of feces samples, 11.3% of environmental, and 2.3% of feed samples. Non-susceptibility profiles included six isolates from feces (14.6%): four non-susceptible isolates (9.8%) to ciprofloxacin, one (2.4%) to nitrofurantoin, and one to both ciprofloxacin and nitrofurantoin (2.4%). Regarding the environmental samples, one profile was non-susceptible to ciprofloxacin (2.4%) and two to nitrofurantoin (4.8%). The serotypes identified included Typhimurium/I4,[5],12:i:-, S. Braenderup/Ohio, S. Newport, S. Anatum/Saintpaul, and S. Westhampton. The epidemiological surveillance of Salmonella and antimicrobial resistance can serve in the creation of strategies for the prevention of the disease and its dissemination throughout the One Health approach.

As safe agents of last resort, carbapenems are reserved for the treatment of infections caused by multidrug-resistant organisms. The impact of β-lactam antibiotics, cefotaxime, and meropenem on the frequency and diversity of carbapenemase-producing organisms recovered from environmental samples has not been fully established. Therefore, this methodological study aimed at determining β-lactam drugs used in selective enrichment and their impact on the recovery of carbapenemase-producing Enterobacterales (CPE) from untreated wastewater. We used a longitudinal study design where 1L wastewater samples were collected weekly from wastewater treatment plant (WWTP) influent and quarterly from contributing sanitary sewers in Columbus, Ohio USA with 52 total samples collected. Aliquots of 500 mL were passed through membrane filters of decreasing pore sizes to enable all the water to pass through and capture bacteria. For each sample, the resulting filters were placed into two modified MacConkey (MAC) broths, one supplemented with 0.5 μg/mL of meropenem and 70 μg/mL of ZnSO 4 and the other supplemented with 2 μg/mL cefotaxime. The inoculated broth was then incubated at 37° C overnight, after which they were streaked onto two types of correspondingly-modified MAC agar plates supplemented with 0.5 μg/mL and 1.0 μg/mL of meropenem and 70 μg/mL of ZnSO 4 and incubated at 37°C overnight. The isolates were identified based on morphological and biochemical characteristics. Then, up to four distinct colonies of each isolate’s pure culture per sample were tested for carbapenemase production using the Carba-NP test. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) MALDI-TOF MS was used to identify carbapenemase-producing organisms. In total 391 Carba-NP positive isolates were recovered from the 52 wastewater samples: 305 (78%) isolates had bla KPC , 73 (19%) carried bla NDM , and 14 (4%) harbored both bla KPC and bla NDM resistance genes. CPE genes of both bla KPC and bla NDM were recovered in both types of modified MAC broths, with 84 (21%) having a bla KPC gene , 22 (6%) carrying bla NDM and 9 (2%) harbored both a bla KPC and bla NDM of isolates recovered from MAC medium incorporated with 0.5ug/mL meropenem and 70ug/mL ZnSO 4 . The most prevalent isolates were Klebsiella pneumoniae , Escherichia coli , and Citrobacter spp .

Aquatic ecosystems are currently facing a multitude of stressors from anthropogenic impacts, including climate change, pollution, and overfishing. Public aquariums positively contribute to ecosystems through conservation, education, and scientific advancement; but may also negatively detract from these systems through collection of animals from the wild and sourcing from commercial suppliers. Changes within the industry have occurred, although evidence-based assessments of 1) how aquariums collect and maintain their populations to determine sustainability of the environment they have harvested; and 2) the welfare of these harvested animals once within the aquariums are still needed. The objectives of this study were to assess the ecosystem health of locations aquariums frequently visit to collect fish from the wild, and then evaluate the wellbeing of fishes at aquariums after extended periods in captivity. Assessments included use of chemical, physical, and biological indicators at field sites, and use of a quantitative welfare assessment at aquariums for comparison to species reared through aquaculture. Anthropogenic pressures at field sites were observed, but no evidence of high degradation or compromised health of animals were found. Welfare assessments of aquarium exhibit tanks produced high-positive scores overall (> 70/84), demonstrating that both wild collected (avg. score 78.8) and aquaculture fishes (avg. score 74.5) were coping appropriately within their environments. Although findings indicated that fish can be taken from the wild at low-moderate rates without any deleterious impact on the environment and cope equally well in aquarium settings, alternatives such as aquaculture should be considered as a strategy to reduce pressure on known stressed aquatic environments or where significant numbers of fishes are being taken.