To date, carbapenem-resistant Enterobacteriaceae have been found predominantly in clinical settings worldwide. Raoultella belongs to the Enterobacteriaceae family which can cause hospital-acquired infections, and carbapenem-resistant Raoultella spp. (CRR) is sporadically reported in the environment. We investigated the distribution and underlying resistance mechanisms of CRR in a wastewater treatment plant (WWTP) from eastern China between January 2018 and February 2019. A total of 17 CRR were isolated from 324 environmental samples, including Raoultella ornithinolytica (n = 15) and Raoultella planticola (n = 2). The detection of CRR was more frequent in the water inlet compared to anaerobic tank, aerobic tank, sludge thickener, activated sludge, mud cake storage area, and water outlet, and CRR was detected in mud cake stacking area. All CRR were resistant to imipenem, meropenem, ampicillin, piperacillin-tazobactam, cefotaxime, ceftazidime, trimethoprim-sulfamethoxazole and fosfomycin. Four different carbapenemase genes were identified, including blaKPC₋₂ (n = 13), blaNDM₋₁ (n = 8), blaNDM₋₅ (n = 1), blaIMP₋₄ (n = 1). Interestingly, isolated R. ornithinolytica from the WWTP were closely related to those reported from human samples in China. Plasmid analysis indicated that IncFII(Yp), IncP6, and IncU mediated blaKPC₋₂ spread, IncX3 and IncN2 mediated blaNDM spread in the environment. The core structure of the Tn3-ISKpn27-blaKPC₋₂-ISKpn6, ISAba125-blaNDM-bleMBL-trpF-dsbD were identified. The study provides evidence that Raoultella spp. may spread alarming carbapenem resistance in the environment and, therefore, the continuous surveillance for carbapenem resistance in the WWTP should be conducted, especially sludge.

The environment is one of the main reservoirs of antibiotic resistance genes (ARGs) but multidrug resistant (MDR) environmental isolates are barely characterised. As suggested by the name, Pedobacter species have been predominantly isolated from soils, but are also recovered from water (including drinking water), chilled food, fish, compost, sludge, glaciers and other extreme environments. The susceptibility phenotype of Pedobacter lusitanus NL19 (isolated from a deactivated uranium mine), its closely related species and the genus type strain were investigated. All strains are MDR bacteria, resistant to β-lactams, colistin, aminoglycosides and ciprofloxacin. Therefore, Pedobacter spp. are likely intrinsically resistant to β-lactams (including ertapenem) and to other three classes of antibiotics. 6%–8% of their total protein-encoding genes encode a diverse collection of putative ARGs, including β-lactamases. These enzymes are highly abundant in all the other Pedobacter strains with sequenced genomes, especially class C, class B3 and class A. LUS-1 and PLN-1 were further characterised in E. coli. LUS-1 is a class A β-lactamase and it conferred an increase in the MIC of cefotaxime, albeit very low. PLN-1 is a class B3 β-lactamase with carbapenemase activity, conferring resistance to ertapenem and a 66x and 16x increase in the MIC of imipenem and meropenem, respectively. PLN-1 also hydrolyses ampicillin, 1st and 3rd generation cephalosporins, and at a lower extent cephamycins and 4th generation cephalosporins. Therefore, Pedobacter spp. encode a large and diverse arsenal of resistance mechanisms that make them environmental superbugs.

The occurrence and distribution of 168 pharmaceutically active compounds (PhACs) in the surface seawater of Jiaozhou Bay (JZB) were investigated using ultra-high-performance liquid chromatography in tandem with a triple-quadrupole mass spectrometer equipped with an electrospray ionization source (UHPLC-ESI-MS-MS). Thirty-six compounds were detected, and 17 of these compounds were first detected in seawater, including sulfabenzamide, sulphacetamide, cephalonium, desacetyl-cefotaxime, cefminox, cefotaxime, cephradine, cefazolin, carprofen, nabumetone, glibenclamide, glimepiride, glipizide, prednisone, fluoromethalone, diazepam and amantadine. The total concentration of PhACs in the surface seawater ranged from 23.6 ng/L to 217 ng/L. The compounds found at the highest mean concentrations included amantadine (24.7 ng/L), lincomycin (8.55 ng/L), carprofen (8.30 ng/L), and tetracycline (7.48 ng/L). The PhAC concentration was higher in the inner bay than in the outside of the bay. In the inner bay, the eastern district showed higher concentrations of PhACs than the western district. Input from the Licun River may be the primary source of pollution. A statistically significant positive correlation was observed between nutrients and PhACs in seawater. Phosphate can be used to indicate the distribution of PhACs in JZB. Based on the individual risk quotient (RQ) values, lincomycin and ofloxacin posed high risks to the relevant aquatic organisms in JZB, especially in the eastern parts. Regular monitoring is required to evaluate the levels of PhACs as they are constantly released into JZB.

The goal of this study was to identify antibiotics with potential risk in river water of the megacity Beijing, China. This was accomplished by using a tiered approach that combined hazard (phase I) and monitoring-based risk (phase II) assessment. Ninety-five candidate antibiotics were screened and 31 was identified as hazardous during phase I assessment. Of these hazardous antibiotics, 29 were identified as persistent and 7 were identified as bioaccumulative antibiotics. Fluoroquinolones, macrolides, sulfonamides, and aminoglycosides account for over 80% of these hazardous antibiotics. During phase II, four antibiotics (erythromycylamine, cefotaxime, ampicillin, and fusidic acid) that were not previously reported were detected in the surface water sampled from four major rivers in Beijing, with concentrations ranging from not detected to approximately 300 ng/L. The ecological risk assessment showed that erythromycylamine, cefotaxime, and ampicillin posed low to high levels of risk to the aquatic organisms. To summarize, erythromycylamine, cefotaxime, and ampicillin were identified as priority antibiotics in rivers in Beijing, China. Our results demonstrated the necessity of conducting monitoring-based verification process in identification of priority antibiotics in a specific region.

Listeria monocytogenes (L. monocytogenes) is frequently detected in ruminants, especially dairy cattle, and associated with the sporadic and epidemic outbreak of listeriosis in farms. In this epidemiological study, the prevalence, virulence, antibiotic resistance profiles, and genetic diversity of L. monocytogenes in three Egyptian dairy cattle farms were investigated. The risk factors associated with the fecal shedding of L. monocytogenes were analyzed. The L. monocytogenes strains from the three farms were categorized into distinct genotypes based on sampling site and sample type through enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR). A total of 1896 samples were collected from animals, environments, and milking equipment in the three farms. Results revealed that 137 (7.23%) of these samples were L. monocytogenes positive. The prevalence of L. monocytogenes in the animal samples was high (32.1%), and the main environmental source of prevalent genotypes in the three farms was silage. For all sample types, L. monocytogenes was more prevalent in farm I than in farms II and III. Risk factor analysis showed seasonal variation in production hygiene. For all sample types, L. monocytogenes was significantly more prevalent in winter than in spring and summer. The level of L. monocytogenes fecal shedding was high likely because of increasing age, number of parities, and milk yield in dairy cattle. Two virulence genes, namely, hlyA & prfA, were also detected in 93 strains, whereas only one of these genes was found in 44 residual strains. Conversely, iap was completely absent in all strains. The strains exhibited phenotypic resistance to most of the tested antibiotics, but none of them was resistant to netilmicin or vancomycin. According to sample type, the strains from the animal samples were extremely resistant to amoxicillin (95.2%, 80/84) and cloxacillin (92.9%, 78/84). By comparison, the strains from the environmental samples were highly resistant to cefotaxime (86.95%, 20/23). Furthermore, 25 multi-antibiotic resistance (MAR) patterns were observed in L. monocytogenes strains. All strains had a MAR index of 0.22–0.78 and harbored antibiotic resistance genes, including extended-spectrum β-lactamase (blaCTX-M [92.7%] and blaDHA-1 [66.4%]), quinolones (qnrS [91.2%], qnrA [58.4%], parC [58.4%], and qnrB [51%]), macrolides (erm[B] [76.6%], erm(C) [1.5%], and msr(A) [27%]), trimethoprim (dfrD [65.7%]), and tetracyclines (tet(M) [41.6%], tet(S) [8%], and int-Tn [26.3%]). ERIC-PCR confirmed that the strains were genetically diverse and heterogeneous. A total of 137 isolated L. monocytogenes strains were classified into 22 distinct ERIC-PCR groups (A–V). Among them, ERIC E (10.2%) was the most prevalent group. These results indicated that environment and milking equipment served as reservoirs and potential transmission ways of virulent and multidrug-resistant L. monocytogenes to dairy animals, consequently posing threats to public health. Silage is the main environmental source of prevalent genotypes on all three farms. Therefore, hygienic measures at the farm level should be developed and implemented to reduce L. monocytogenes transmission inside dairy cattle farms.

Marine fish species were analyzed for culturable and total metagenomic microbial diversity, antibiotic resistance (AR) pattern, and horizontal gene transfer in culturable microorganisms. We observed a high AR microbial load of 3 to 4 log CFU g⁻¹. Many fish pathogens like Providencia, Staphylococcus, Klebsiella pneumoniae, Enterobacter, Vagococcus, and Aeromonas veronii were isolated. Photobacterium and Vibrio were two major fish and human pathogens which were identified in the fish metagenome. Other pathogens that were identified were Shewanella, Acinetobacter, Psychrobacter, and Flavobacterium. Most of these pathogens were resistant to multiple antibiotics such as erythromycin, kanamycin, neomycin, streptomycin, penicillin, cefotaxime, bacitracin, rifampicin, trimethoprim, ciprofloxacin, and doxycycline with a high multiple antibiotic resistance index of 0.54–0.77. The fish microflora showed high prevalence of AR genes like bla TEM, Class I integron, tetA, aph(3′)-IIIa, ermB, aadA, and sul1. Nineteen of 26 AR isolates harbored Class I integrons showing high co-resistance to trimethoprim, kanamycin, doxycycline, and cefotaxime. Mobile R-plasmids from 6 of the 12 AR pathogens were transferred to recipient E. coli after conjugation. The transconjugants harbored the same R-plasmid carrying bla CTX₋M, dfr1, tetA, bla TEM, and cat genes. This study confirms that fish is a potential carrier of AR pathogens which can enter the human gut via food chain. To the best of our knowledge, this is the first study in the Indian subcontinent reporting a direct evidence of spread of AR pathogens to humans from specific marine fish consumption.

The occurrence, distribution, and risk assessment of antibiotics in freshwater systems are receiving global attention, because of their impact on the environment and human health. However, few studies have focused on this topic in Northeast China and its Songhua River, the third-largest river in China. This study investigated the occurrence and distribution of 12 antibiotics, including three cephalosporins (cefazolin, cefmetazole, cefotaxime), three macrolides (azithromycin, clarithromycin, roxithromycin), three fluoroquinolones (ofloxacin, norfloxacin, flumequine), and three sulfonamides (sulfadiazine, sulfapyridine, sulfamethoxazole) in the mainstream and tributaries of the Songhua River. A total of 152 surface water samples were collected in January, May, July, and October 2016. These samples were analyzed using solid-phase extraction (SPE) and ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). The results indicated the wide use of all 12 antibiotics in the mainstream of the Songhua River. Sulfamethoxazole and cefazolin were the dominant antibiotics, with maximum concentrations of 73.1 and 65.4 ng L⁻¹, respectively. Other antibiotics were present at mean concentrations below 15 ng L⁻¹, except cefmetazole, present at a mean concentration of 35.6 ng L⁻¹. The spatial distribution of antibiotics showed that unbalanced regional development may lead to the distribution pattern of the antibiotics in the tributaries and the mainstream. Thus study also assessed the seasonal variation of antibiotics in urban surface water; cephalosporin, sulfonamide, fluoroquinolone, and macrolide concentrations were significantly higher during the icebound season than during non-icebound season. This may be due to the low temperature and water flow of the river in winter. Risk assessment showed that azithromycin, clarithromycin, roxithromycin, flumequine, and sulfamethoxazole posed a low or median risk to the aquatic organisms in the mainstream. The potential risks created by antibiotics to the aquatic environment should not be neglected in the Songhua River. The potential risks created by antibiotics to the aquatic environment should not be neglected in the Songhua River.

This study aimed to assess the antibiogram of Aeromonas strains recovered from cattle faeces and the potential pathogenic status of the isolates. The antibiogram of the Aeromonas isolates demonstrated total resistance to clindamycin oxacillin, trimethoprim, novobiocin and ticarcillin. However, Aeromonas strains were sensitive to cefotaxime, oxytetracycline and tobramycin. The Aeromonas strains from Lovedale and Fort Cox farms were found to possess some virulence genes. The percentage distribution was aer 71.4 %, ast 35.7 %, fla 60.7 %, lip 35.7 % and hlyA 25 % for Lovedale farm and aer 63.1 %, alt 10.5 %, ast 55.2 %, fla 78.9 %, lip 21 % and hlyA 35.9 % for Fort Cox farm. Class 1 integron was present in 27 % of Aeromonas isolates; the bla TEM gene was present in 34.8 %, while the blaP1 class A β-lactamase gene was detected in 12.1 % of the isolates. Approximately 86 % of the isolates formed a biofilm on microtitre plates. The presence of multiple antibiotic resistance and virulence genes in Aeromonas isolates from cattle faeces reveals the pathogenic and infectious importance of these isolates and is of great significance to public health. The possession of a biofilm-forming capability by such isolates may lead to difficulty during the management of infection related to Aeromonas species.

Aeromonas species are broadly distributed in nature and agricultural environments and have been isolated from feces, bedding, and drinking water of healthy pigs. We assessed the incidence, virulence properties, and antimicrobial resistance profile of Aeromonas spp., isolated from pig feces. Antibiogram was done using the disc diffusion methods, and polymerase chain reaction was used for the detection of putative virulence genes. Identification of isolates revealed three phenotypic species with percentage distribution as follows: Aeromonas hydrophila 23 (45.1 %), Aeromonas caviae 16 (31.4 %), and Aeromonas sobria 12 (23.5 %). All Aeromonas isolates in the study were absolutely susceptible to cefotaxime and resistant to penicillin. A. cavaie and A. sobria demonstrated absolute susceptibility against ciprofloxacin and streptomycin. Aeromonas species showed varied susceptibility to cephalothin as follows: A. hydrophila 78.3 %, A. cavaie 93.7 %, and A. sobria 91.7 %. The percentage distribution of virulence genes among Aeromonas isolates were as follows: Aerolysin (aer) 74.5 %, flagellin gene (fla) 68.6 %, cytotoxin (hly A) 43.1 %, lipase (lip) 39.2 %, enterotoxic activities (ast) 31.3 %, and cytotonic gene (alt) 13.7 %. Reports from this study shows that Aeromonas species isolated from pig fecal samples are multi-drug resistant and possess virulence potential which may result to possible risk of human or animal infection and likely contamination of food and water from this sources.