The objective of this study was to determine the prevalence and characteristics of antimicrobial-resistant Enterococcus faecalis and E. faecium isolated from the oral cavities of captive giant pandas in China. The virulence-associated determinant and antimicrobial resistance genes were detected and antimicrobial susceptibility tests were performed on 54 strains of each bacterium. All isolates showed 100% multidrug resistance. E. faecalis isolates showed a higher percentage of strains resistant to gentamicin (48.1%), vancomycin (55.6%), linezolid (100%), and streptomycin (33.3%) than E. faecium isolates. The resistance genes of Enterococcus spp. were present to highly varying extents according to antibiotic type, their presence breaking down for E. faecalis and E. faecium respectively as aac(6')/aph(2″) 5.56% and 5.56%; aph(3')-Ⅲ 0% and 14.81%; ant(6)-I 0% and 3.7%; ant(4')-Ia 0% and 64.81%; tetL 20.37% and 100%; vanA 92.59% and 46.3%; vanB 0% and 0%; cfr 0% and 90.74%; optrA 96.3% and 3.7%; blaZ 0% and 1.85%; blaTEM 0% and 0%; tetA 20.37% and 0%; tetC 24.07% and 100%; tetM 0% and 0%; ermA 12.96% and 100%; ermB 5.56% and 3.7%; and ermC 0% and 1.85%.Virulence-associated determinants were detected in this research, which typically include efaA, gelE, asa1, ace, cylA, esp and hyl; however, the latter three were not detected. High proportions of the isolates carried the efaA, gelE, asa1, and ace genes. Respectively for E. faecalis and E. faecium their detection was efaA 98.1% and 85.2%; gelE 98.1% and 87%; asa1 92.6% and 87%; and ace 87% and 85.2%. This is the first study on the potential disease risk and antimicrobial-resistant characteristics of E. faecalis and E. faecium isolates in giant panda oral cavities. The results of this study show that the antimicrobial resistance rate of Enterococcus spp. isolated from the oral cavity of captive pandas is very high, and thus needs to be monitored.

The value of neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (Kim-1), and liver-type fatty acid binding protein (L-FABP) was assessed in early diagnosis of gentamicin-induced acute kidney injury (AKI) in dogs. Subcutaneous gentamicin injection in 16 healthy adult beagles made the AKI model. Blood was sampled every 6 h to detect NGAL, Kim-1, L-FABP, and serum creatinine (SCr) concentrations. Kidney tissue of two dogs was taken before the injection, as soon as SCr was elevated (78 μmol/L), and when it had risen to 1.5 times the baseline, and haematoxylin-eosin staining and transmission electron microscopy (TEM) were used to observe changes. NGAL, Kim-1, and SCr levels were significantly increased (P < 0.05) at 18, 30, and 78 h post injection, but L-FABP concentration was not associated with renal injury. At the earliest SCr elevation stage, findings were mild oedema, degeneration, and vacuolisation in renal tubular epithelial cells in pathology, and mild cytoplasmic and mitochondrial oedema in TEM. At this time point, NGAL and Kim-1 concentrations were significantly increased (P < 0.05), indicating that these two molecules biomark early kidney injury in dogs. Using receiver operating characteristic curve analysis, their warning levels were > 25.31 ng/mL and > 48.52 pg/mL. Plasma NGAL and Kim-1 above warning levels are early indicators of gentamicin-induced AKI in dogs.

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) is a highly resistant and difficult to cure zoonotic microorganism, which makes up a large part of food toxic infections and has shown high prevalence among pig population all over the world. The aim of the study was to establish the occurrence of MRSA in slaughterhouses, evaluate its antimicrobial resistance, and verify whether there are any differences or similarities with reference to other European countries. Material and Methods: A total of 100 pigs, 105 carcasses, 19 workers, and 24 samples from the environment of several slaughterhouses were examined by conventional microbial and molecular methods. Results: In total, 78 MRSA isolates were found. MRSA prevalence in slaughtered pigs varied from 8.0% to 88.6% depending on the slaughterhouse, reaching higher prevalence in slaughterhouses with higher slaughter capacity. In total, 21.1% of all workers were carriers of MRSA and 6.7% of carcasses were contaminated with MRSA. The 98.2% of MRSA isolates were resistant to penicillin, 89.1% to tetracycline, 60.1% to erythromycin, 65.5% to gentamycin, and 15 different spa types were found, among which spa type t01333 was most widespread. Conclusion: The study indicated that MRSA prevalence and spa types differed according to slaughterhouse slaughter capacity and good hygiene practices. Quite high MRSA occurrence among slaughterhouse workers is one of the main factors which increase pork contamination risk.

Enterococci are widespread, being part of the bacterial flora of humans and animals. The food chain can be therefore considered as the main route of transmission of antibiotic resistant bacteria between the animal and human populations. Milk in particular represents a source from which resistant bacteria can enter the human food chain. The aim of the study was to determine the occurrence and resistance to antimicrobial agents of Enterococcus spp. strains isolated from raw goat’s milk samples. A total of 207 goat’s milk samples were collected. Samples were cultivated on selective media and confirmed as E. faecium or E. faecalis and screened for selected resistance genes by PCR. Drug susceptibility determination was performed by microdilution on Sensititre EU Surveillance Enterococcus EUVENC Antimicrobial Susceptibility Testing (AST) Plates and Sensititre US National Antimicrobial Resistance Monitoring System Gram Positive CMV3AGPF AST Plates. Enterococcal strains totalling 196 were isolated, of which 40.8% were E. faecalis and 15.3% were E. faecium. All tested isolates were susceptible to linezolid, penicillin and tigecycline. For most other antimicrobials the prevalence of resistance was 0.5–6.6% while high prevalence of quinupristin/dalfopristin (51.5%), tetracycline (30%) and lincomycin (52%) resistance was observed. This study affords better knowledge concerning the safety of raw goat’s milk in terms of the enterococci possible to isolate from this foodstuff. It seems that enterococci in milk are still mostly susceptible to antimicrobials of major concern as multiply resisted drugs, such as gentamycin and vancomycin. However, the presence of multi-resistant strains in goat milk is cause for apprehension.

Land application of manure that contains antibiotics and resistant bacteria may facilitate the establishment of an environmental reservoir of antibiotic-resistant microbes, promoting their dissemination into agricultural and natural habitats. The main objective of this study was to search for acquired antibiotic resistance determinants in the gut microbiota of wild boar populations living in natural habitats. Gastrointestinal samples of free-living wild boars were collected in the Zemplén Mountains in Hungary and were characterised by culture-based, metagenomic, and molecular microbiological methods. Bioinformatic analysis of the faecal microbiome of a hunted wild boar from Japan was used for comparative studies. Also, shotgun metagenomic sequencing data of two untreated sewage wastewater samples from North Pest (Hungary) from 2016 were analysed by bioinformatic methods. Minimum spanning tree diagrams for seven-gene MLST profiles of 104 E. coli strains isolated in Europe from wild boars and domestic pigs were generated in Enterobase. In the ileum of a diarrhoeic boar, a dominant E. coli O112ab:H2 strain with intermediate resistance to gentamicin, tobramycin, and amikacin was identified, displaying sequence type ST388 and harbouring the EAST1 toxin astA gene. Metagenomic analyses of the colon and rectum digesta revealed the presence of the tetQ, tetW, tetO, and mefA antibiotic resistance genes that were also detected in the gut microbiome of four other wild boars from the mountains. Furthermore, the tetQ and cfxA genes were identified in the faecal microbiome of a hunted wild boar from Japan. The gastrointestinal microbiota of the free-living wild boars examined in this study carried acquired antibiotic resistance determinants that are highly prevalent among domestic livestock populations.

The value of neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (Kim-1), and liver-type fatty acid binding protein (L-FABP) was assessed in early diagnosis of gentamicin-induced acute kidney injury (AKI) in dogs.

The aim of this study was to evaluate the occurrence of Shiga toxin (stx)-producing Escherichia coli (STEC) in diarrheic newborn calves, as well as the resistance profile of this microorganism against antimicrobials routinely used in veterinary therapy. The antimicrobial profile of Eugenia uniflora against E. coli clinical isolates was also analyzed. Specimens from the recto-anal junction mucosa were investigated by using chromogenic medium and identification of E. coli was done using microbiological methods (Gram staining, indole test, methyl red test, Voges-Proskauer test, citrate test, urease test, and hydrogen sulfide test). The stx1 and stx2 genes corresponding to the STEC pathotype were evaluated by using polymerase chain reaction and electrophoresis. The susceptibility profile to antimicrobial agents commonly used in veterinary therapeutic practice and the antimicrobial effect of lyophilized hydroalcoholic extract of E. uniflora L. leaves against E. coli clinical isolates were evaluated by disk diffusion and microdilution methods. Shiga toxin-positive E. coli was identified in 45% of diarrheic newborn calves (stx1 = 23.2%, stx2 = 4.0%, stx1 + stx2 = 18.2%). The frequency of stx-positive E. coli in the bacterial population was equal to 17.0% (168/990 clinical isolates): 97 (9.8%) stx1-positive E. coli, 12 (1.2%) stx2-positive E. coli, and 59 (6.0%) stx1 + stx2-positive E. coli isolates. All stx-positive E. coli analyzed showed resistance to multiple drugs, that is, from 4 to 10 antimicrobials per clinical isolate (streptomycin, tetracycline, cephalothin, ampicillin, sulfamethoxazole + trimethoprim, nitrofurantoin and nalidixic acid, ciprofloxacin, gentamicin, and chloramphenicol). Effective management measures should be implemented, including clinical and laboratory monitoring, in order to promote animal and worker health and welfare, prevent and control the spread of diseases, and ensure effective treatment of infectious diseases. The E. uniflora L. leaves showed inhibition of microbial growth based on the diameter of halos, ranging from 7.9 to 8.0 mm and 9.9 to 10.1 mm for concentrations of 50 and 150 mg/mL, respectively. This plant displayed bacteriostatic action and a minimum inhibitory concentration of 12.5 mg/mL for all clinical isolates. Its clinical or synergistic effects with antimicrobial agents must be determined from clinical and preclinical trials.

Pseudomonas aeruginosa is an important animal pathogen and contributes to hemorrhagic pneumonia in mink. Between April 2011 and December 2016, samples of lung, liver, and spleen were collected from mink with this disease on 11 mink farms in 5 Chinese provinces. From these samples, we obtained 98 isolates of P. aeruginosa that belonged to 5 serotypes: G (n = 58), I (n = 15), C (n = 8), M (n = 5), and B (n = 2); 10 isolates were not typeable (10/98). More than 90% of the isolates formed biofilms, and 85% produced slime. All 98 isolates were resistant to 10 antibiotics (oxacillin, ampicillin, penicillin G, amoxicillin, ceftriaxone, cefazolin, cefaclor, tilmicosin, tildipirosin, and sulfonamide). However, almost all were susceptible to gentamicin, polymyxin B, and amikacin. We identified 56 unique genotypes by pulsed-field gel electrophoresis. These findings have revealed genetic diversity and high antimicrobial resistance in P. aeruginosa isolated from mink with hemorrhagic pneumonia and will facilitate the prevention and control of this disease.

The objective of this study was to estimate the presence of the important foodborne pathogen Campylobacter jejuni in organically raised chickens in the province of Quebec. The recovered isolates were further characterized for their antimicrobial resistance profile, autoagglutination property and chemotaxis. Antimicrobial resistance was evaluated using agar dilution for: tetracycline, erythromycin, chloramphenicol, ciprofloxacin, gentamicin, nalidixic acid, clindamycin, ampicillin, azithromycin, bacitracin, and ceftiofur. Autoagglutination was measured by monitoring optical density changes in a bacterial suspension after 3 h of incubation at room temperature. Chemotaxis was evaluated after a contact time of 3 h between isolates and mucin, using a quantitative protocol. A total of 10 lots of chickens was sampled in August and September 2009; half of them were positive for the presence of C. jejuni. Antimicrobial resistance was found only for tetracycline (44%), erythromycin (6%), azithromycin (6%) and clindamycin (2%). Variation was observed in the minimum inhibitory concentrations (MICs) for ceftiofur and bacitracin, for which C. jejuni possess intrinsic resistance. Autoagglutination and chemotaxis varied among isolates and lot-level differences in these were observed. Autoagglutination and chemotaxis levels appeared as independent isolate properties. Further monitoring and characterization of isolates originating from organic chickens is of interest since this type of production might represent another source of exposure of consumers to a variety of the foodborne pathogen C. jejuni.

In 8 Holstein cows, 50 colony-forming units (CFU) Of Escherichia coli was administered into 1 mammary gland. Infections were established in all inoculated glands. In 4 of the 8 cows, 500 mg of gentamicin sulfate was administered by intramammary infusion 14 hours after inoculation; the other 4 cows were untreated controls. Infusions of gentamicin also were given after each of the 3 successive milkings after the initial infusion, so that a total dose of 2 g of gentamicin was given to each of the treated cows. During the 33-hour treatment period and for the first milking after the last infusion of gentamicin, the treated cows had a mean gentamicin concentration of greater than or equal to 31.0 microgram/ml in milk samples that were collected from inoculated quarters immediately before each milking. Concentrations of 0.34 and 0.69 microgram of gentamicin were detected in milk from 2 cows at 8 days after inoculation with E coli. Mean serum concentrations of gentamicin were > 0.37 microgram/ml throughout the treatment period and the first 12 hours after the last infusion, with a mean peak concentration of 0.96 microgram/ml at 24.4 hours. The range of peak concentration of gentamicin detected in urine from all treated cows was 42 to 74.4 microgram/ml. Peak concentration of E coli in milk in the treated cows (6.08 +/- 1.02 log10 CFU/ml) did not significantly (P > 0.05) differ from that of the control cows (5.26 +/- 1.00 log10 CFU/ml). Similarly, mean duration of infection in the treated cows (54 hours) did not differ significantly from that of the control cows (48 hours). The treatment groups also did not differ significantly in peak concentrations of albumin or IgG1 in milk, although mean concentrations of albumin and IgG1 at 16 hours after inoculation, and of albumin at 20 hours, was significantly (P < 0.05) higher in the milk from control cows than from the treated cows. Mean values of peak rectal temperature and of mean rectal temperature throughout the trial did not differ between the groups. At the end of the 4-week trial, 1 of 4 inoculated glands in treated cows and 3 of 4 in control cows had somatic cell counts less than or equal to preinoculation concentrations (5.18 log10 cells/ml). Intramammary administration of gentamicin did not affect the duration or severity of experimentally induced E coli mastitis. In addition, substantial concentrations of gentamicin were detected in the serum of treated cows, suggesting that intramammary treatment may result in prolonged drug residues in tissue.