BACKGROUND: Escherichia coli is a particularly complex species that is grouped into pathotypes of partly zoonotic intestinal pathogenic E. coli and extraintestinal pathogenic E. coli (ExPEC). Strains belonging to ExPEC are able to cause various clinical signs in hosts and due to similar genetic determinants, these hosts may act as a source of infection for each other. OBJECTIVES: Recent reports of outbreaks of human urinary tract infections (UTIs) have stimulated interest in the potential that E. coli from animals has to cause human UTIs via the food supply especially poultry meat, so we aimed to assess the genetic relationships between strains from these two hosts. METHODS: A total  of  260  E. coli  isolates  were  obtained  from human  UTI’s (160 strains) and  poultry colibacillosis cases (100 strains)  and  phylogenetic  grouping  was  done  based  on  the Triplex-PCR  method and virulence genotyping was carried out using a modified Tetraplex-PCR detecting hly, iucD, papEF and sfa/focDE genes. RESULTS: Statistical analysis of results demonstrated that prevalence of B2 & D phylogroups in human UTI’s (77%) and D & A groups in poultry strains (66%) are higher than others, considerably. Statistical analysis showed that distribution of A phylogroup within poultry isolates versus human and B2 phylogroup within human isolates versus poultry ones were higher, significantly. It was shown that iucD is noticeablymore prevalent in poultry strains rather than human isolates,. Also, sfa/focDE gene was significantly more distributed in human strains than poultry isolates. CONCLUSIONS: In sum, despite the minor genetic differences between isolates from both hosts, our results showed that there are major genetic similarities in E. coli isolates from human UTI and poultry colibacillosis cases in the region and these two hosts can play an important role as infection source for the other one.   ________________________________________________________________

The experiment evaluated the effects of intravenous administration of polymyxin B on experimental endotoxaemia in sheep. Twenty clinically healthy fat-tailed sheep were randomly divided into: a group treated with 6,000 U/kg of polymyxin B, a group at 12,000 U/kg, and positive and negative controls. Endotoxaemia was induced by intravenous administration of lipopolysaccharide (LPS) from E. coli serotype O55:B5 at 0.5 μg/kg. polymyxin was infused intravenously along with 2.5 L of isotonic intravenous fluids at 20 mL/kg/h. The positive control group received LPS and 2.5 L of isotonic fluids, the negatives receiving just 2.5 L of isotonic fluids. Clinical signs were evaluated before and at 1.5, 3, 4.5, 6, 24, and 48 h after LPS administration. Blood was also sampled at the denoted hours and serum haptoglobin, tumour necrosis factor-α (TNF-α), and plasma lactate concentrations were assayed. The serum concentration of TNF-α in the positive control group increased significantly up to 48 h after LPS administration. The concentration of TNF-α was significantly different from those of the polymyxin B and positive control groups from 3 to 48 h; also, the concentrations of haptoglobin at different times in the polymyxin groups were lower than those of the positive control group and were significant at hours 3 to 48 (P < 0.05). Following the LPS administration, haptoglobin and TNF-α concentrations changed without significant difference between the two polymyxin B groups. Polymyxin B (6,000 U/kg) restrained blood lactate concentrations. Furthermore, it significantly improved the clinical signs in endotoxaemic animals, including rectal temperature and heart and respiratory rates. Polymyxin B may be an antiendotoxic in fat-tailed sheep.

The purpose of this study was to determine the microbiological quality of food fish and its safety for consumers. The study included 24 fish representing grass carp, bighead carp, Siberian sturgeon, and wels catfish. Specimens were collected in winter. Aerobic bacteria, psychrophilic, Enterobacteriaceae, Staphylococcus spp., and E. coli counts were made, and the presence of Salmonella spp., L. monocytogenes, S. aureus, and other coagulase-positive staphylococci was investigated. The microbiological analysis showed a similar level of aerobic, psychrophilic, and Staphylococcus spp. contamination of the four fish species. The Enterobacteriaceae count was higher in the muscles of grass carp and bighead carp than S. sturgeon and wels catfish. No pathogenic bacteria such as Salmonella spp., E. coli, L. monocytogenes, Staphylococcus aureus, or other coagulase positive staphylococci were found in samples of the examined fish species. The fresh fish examined in this study were of good microbiological quality and there was no health risk for consumers.

Objective: The present study was carried out to identify the associated Salmonella and Escherichia coli in betel leaves (Piper betle), and to develop an effective method to remove those microbes. Materials and methods: Betel leaves were collected from local and whole sale markets, and borouj (cultivation place). Salmonella and E. coli were isolated and identified by cultural, morphological, and biochemical tests followed by confirmation by polymerase chain reaction (PCR) targeting the genus specific 16S rRNA genes. Antibiogram of the isolated bacteria was performed by disc diffusion method. Different concentrations of Salmosan-A Soln were used to remediate the contaminating bacteria keeping the quality of betel leaves for longer periods.Results: Total Salmonella counts in the betel leaves were 3.9×105, 4.9×106, 3.5×104, 1.1×103 and 1.5×103 CFU/mL, while E. coli counts were 5.5×107, 6.3×107, 4.4×105, 3.3×103 and 3.1×103 CFU/mL in the betel leaves collected from K.R. market, Kewatkhali Bazaar, whole sale market, borouj in Kushtia and borouj in Natore, respectively. Antibiogram study revealed that the isolated bacteria were sensitive to doxycyclline, ciprofloxacin, chloramphenicol and cefotaxime. Application of 0.3% Salmosan-A Soln was found to be the most effective and suitable, where [J Adv Vet Anim Res 2018; 5(1.000): 73-80]

Poultry has been identified as a reservoir of foodborne enteric pathogens and antimicrobial resistant bacteria. The objective of this study was to describe and compare antimicrobial resistant isolates from an Ontario broiler chicken farm-level baseline project (2003 to 2004) to the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) Ontario abattoir and retail surveillance data from 2003, and to the most recent (2015) CIPARS Ontario chicken surveillance data in order to assess the impact of an industry-wide policy change in antimicrobial use. Ceftiofur resistance (TIO-R) prevalence in Salmonella decreased by 7% on farm between 2003 and 2004 and 2015. During the same timeframe, TIO-R E. coli prevalence decreased significantly by 16%, 11%, and 8% in farm, abattoir, and retail samples, respectively. Gentamicin resistant (GEN-R) E. coli, however, increased by 10% in farm and 15% in retail-derived isolates, and trimethoprim-sulfamethoxazole resistant (TMSm-R) E. coli increased significantly by 20%, 18%, and 5% in farm, abattoir, and retail isolates, respectively. Similarly, ciprofloxacin-resistant (CIP-R) Campylobacter spp. significantly increased in retail isolates by 11% and increased in farm (33%) and abattoir isolates (7%). The decrease in TIO-R Salmonella/E. coli in recent years is consistent with the timing of an industry-led intervention eliminating the preventive use of ceftiofur, a third generation cephalosporin and class of antimicrobials deemed critically important to human medicine. The rise in GEN-R and TMSm-R prevalence is indicative of recent shifts in antimicrobial use. Our study highlights the importance of integrated surveillance in detecting emerging trends and determining the efficacy of interventions to improve food safety.

Colistin is one of the last-resort antibiotics for the treatment of multidrug-resistant infections in humans, but transmissible colistin-resistance genes have emerged in bacteria from animals. The rapid and sensitive detection among animals of colonization with bacteria carrying these genes is critical in helping to control further spread. Here we describe a method for broth enrichment of colistin-resistant Escherichia coli from animal fecal and cecal samples followed by real-time polymerase chain reaction (PCR) for the simultaneous detection of two of the main colistin-resistance genes, mcr-1 and mcr-2. The PCR uses a single set of nondegenerative primers, and mcr variants can be differentiated by melt-curve analysis. Overnight culture enrichment was effective for amplifying colistin-resistant E. coli, even when initially present in numbers as low as 10 bacteria per gram of sample. The mcr-1 and mcr-2 genes were not found in any of the Ontario swine and poultry samples investigated.

OBJECTIVE To determine the in vitro effects of calcitriol on indicators of immune system function in blood samples collected from healthy dogs. SAMPLE Blood samples from 8 healthy adult dogs. PROCEDURES Blood samples were incubated with calcitriol (10(−7)M) or control substance for 24 hours. Afterward, lipopolysaccharide (LPS)-, lipoteichoic acid (LTA)-, and N-acetylmuramyl-l-alanyl-d-isoglutamine hydrate (MDP)-stimulated leukocyte production of tumor necrosis factor (TNF) and interleukin-10 (IL10) were measured with a canine-specific multiplex assay. Phagocytosis of opsonized Escherichia coli and leukocyte expression of constitutive toll-like receptor 4 (TLR4) were evaluated via flow cytometry. Blood samples from 3 dogs were used to create a concentration-response curve to evaluate whether the observed cytokine modulation was concentration dependent. RESULTS Incubation of canine blood samples with calcitriol resulted in significant decreases in LPS-, LTA-, and MDP-stimulated leukocyte production of TNF but not IL10. Blunting of TNF production was concentration dependent. Leukocyte calcitriol exposure had no significant effect on phagocytosis and TLR4 expression. CONCLUSIONS AND CLINICAL RELEVANCE These data indicated that calcitriol induced an anti-inflammatory shift in canine leukocytes exposed to LPS, LTA, and MDP in vitro, without altering phagocytosis or TLR4 expression. Thus, calcitriol could represent a novel candidate immunomodulatory treatment for dogs.

Forty-nine fresh goat’s milk samples produced by local farmers and sold in market for public consumption as well as raw goat milk in Johor, Malaysia were analysed for total plate count(TPC) , E. coli, Coliform, Brucella melitensis, Brucella abortus,Coxiella burnetii as well as aflatoxin M1 (AFM1) content, as measures for food safety. The mean counts per ml for TPC were 4.90 x 105, 6.50 x 105, 1.60 x 105 and 1.48 x 106 for pasteurised, unpasteurised and unknown (status of pasteurisation) milk sold in the market as well as the raw milk from milkcollection center (MCC), respectively. Among pasteurised samples, only one had TPC count higher than the permitted level whereas the rest were all within the permitted level. The mean counts per ml for E. coli were <1.00 x 102 for pasteurised and unknown milkwhereas 1.67 x 101 for unpasteurised and 1.18 x 102 for raw milk. The mean counts per ml for coliform were 9.53 x 103, 9.76 x103, 1.20 x 102 and 1.16 x 104 for pasteurised, unpasteurised, unknown milk and raw milk, respectively. Overall, no significantdifferences on the bacterial counts in both pasteurised and unpasteurised milk. All milk samples were negative of B. melitensis and B. abortus, but one unknown sample fromthe market and two raw samples from MCC were positive of C. burnetii through the ELISA test. The unknown sample from the market showed the presence of C. burnetii when further analysed microscopically. Meanwhile, no sample exceeded the permitted level of AFM1 in milk.

Septicaemic pasteurellosis is a fatal, sometimes epidemic, bacterial disease of domestic and wild animals including deer, bison, elk, and pronghorn antelope caused by Pasteurella multocida. This is the case report of septicaemic pasteurellosisin a captive sambar deer. The carcass was sent from Royal Endurance Stable, Bachok, Kelantan to the Kota Bharu RegionalVeterinary Laboratory for post-mortem. Gross examination of organs was followed by collection of specimens from lung, kidney,liver, spleen and heart for histopathology and bacterial examination. Pooled organ samples with rumen content were collected and sent to the nearest Chemistry Department for investigation. For histology, the liver, lung, spleen, kidney, and heart specimens were fixed in 10% neutral formalin, and routinely embedded in paraffin. Fivemicrometer sections were stained with H&E. Other tests such as worm and ectoparasiteidentification were conducted to identify the parasites. Post-mortem lesions revealed generalised haemorrhage in the organs.Pasteurella multocida serogroup B and E. coli were isolated from multiple tissues of the animal. Histological examination alsorevealed severe congestion and haemorhage of multiple tissues with infiltration of the inflammatory cells. The most likely mode of transmission of these bacteria is through an infected wound and into the bloodstream, thereby causing severe septicemia and death to the animal.

OBJECTIVE To evaluate a fluorescence resonance energy transfer quantitative PCR (FRET-qPCR) assay for detection of gyrA mutations conferring fluoroquinolone resistance in canine urinary Escherichia coli isolates and canine urine specimens. SAMPLE 264 canine urinary E coli isolates and 283 clinical canine urine specimens. PROCEDURES The E coli isolates were used to validate the FRET-qPCR assay. Urine specimens were evaluated by bacterial culture and identification, isolate enrofloxacin susceptibility testing, and FRET-qPCR assay. Sensitivity and specificity of the FRET-qPCR assay for detection of gyrA mutations in urine specimens and in E coli isolated from urine specimens were computed, with results of enrofloxacin susceptibility testing used as the reference standard. RESULTS The validated FRET-qPCR assay discriminated between enrofloxacin-resistant and enrofloxacin-susceptible E coli isolates with an area under the receiver operating characteristic curve of 0.92. The assay accurately identified 25 of 40 urine specimens as containing enrofloxacin-resistant isolates (sensitivity, 62.5%) and 226 of 243 urine specimens as containing enrofloxacin-susceptible isolates (specificity, 93.0%). When the same assay was performed on E coli isolates recovered from these specimens, sensitivity (77.8%) and specificity (94.8%) increased. Moderate agreement was achieved between results of the FRET-qPCR assay and enrofloxacin susceptibility testing for E coli isolates recovered from urine specimens. CONCLUSIONS AND CLINICAL RELEVANCE The FRET-qPCR assay was able to rapidly distinguish between enrofloxacin-resistant and enrofloxacin-susceptible E coli in canine clinical urine specimens through detection of gyrA mutations. Therefore, the assay may be useful in clinical settings to screen such specimens for enrofloxacin-resistant E coli to avoid inappropriate use of enrofloxacin and contributing to antimicrobial resistance.