Caecal Samples Research Articles

A muramidase from Acremonium alcalophilum hydrolyse peptidoglycan found in the gastrointestinal tract of broiler chickens.

This study evaluates peptidoglycan hydrolysis by a microbial muramidase from the fungus Acremonium alcalophilum in vitro and in the gastrointestinal tract of broiler chickens. Peptidoglycan used for in vitro studies was derived from 5 gram-positive chicken gut isolate type strains. In vitro peptidoglycan hydrolysis was studied by three approaches: (a) helium ion microscopy to identify visual phenotypes of hydrolysis, (b) reducing end assay to quantify solubilization of peptidoglycan fragments, and (c) mass spectroscopy to estimate relative abundances of soluble substrates and reaction products. Visual effects of peptidoglycan hydrolysis could be observed by helium ion microscopy and the increase in abundance of soluble peptidoglycan due to hydrolysis was quantified by a reducing end assay. Mass spectroscopy confirmed the release of hydrolysis products and identified muropeptides from the five different peptidoglycan sources. Peptidoglycan hydrolysis in chicken crop, jejunum, and caecum samples was measured by quantifying the total and soluble muramic acid content. A significant increase in the proportion of the soluble muramic acid was observed in all three segments upon inclusion of the microbial muramidase in the diet.

Characterization of faecal and caecal microbiota of free-ranging black-tailed prairie dogs (Cynomys ludovicianus) using high-throughput sequencing of the V4 region of the 16S rRNA gene.

Black-tailed prairie dogs (Cynomys ludovicianus) are keystone species within their grassland ecosystems; their population stability affects a multitude of other species. The goals of this study were to explore, describe and compare the bacterial communities in caecal and hard faecal samples from free-ranging black-tailed prairie dogs (n = 36) from KS, USA, using high-throughput sequencing of the V4 region of the 16S rRNA gene and to compare sex and geographic locations. A total of 22 paired faecal and caecal samples were collected post-mortem from free-ranging black-tailed prairie dogs from 5 different geographical locations. The results revealed that the microbiota of both faecal and caecal samples were dominated by the phylum Firmicutes (genera belonging to the Clostridiales order). There was significantly greater richness in faecal compared with caecal samples. There were significant differences between the 5 different geographic regions (P < 0.001), specifically in the relative abundances of genera. There were differences in rare members of the microbiome between faecal samples from male and female prairie dogs but with no significant impact on overall community structure. This study provides novel data and expands our knowledge about the gastrointestinal microbiome composition of free-ranging black-tailed prairie dogs, which has potential to inform conservation efforts and improve their captive management.

Abattoir-based study of Salmonella prevalence in pigs at slaughter in Great Britain

Abstract Consumption of pork and pork products can be associated with outbreaks of human salmonellosis. Salmonella infection is usually subclinical in pigs, and farm-based control measures are challenging to implement. To obtain data on Salmonella prevalence, samples can be collected from pigs during the slaughter process. Here we report the results of a Great Britain (GB) based abattoir survey conducted by sampling caecal contents from pigs in nine British pig abattoirs during 2019. Samples were collected according to a randomised stratified scheme, and pigs originating from 286 GB farms were included in this survey. Salmonella was isolated from 112 pig caecal samples; a prevalence of 32.2% [95% confidence interval (CI) 27.4–37.4]. Twelve different Salmonella serovars were isolated, with the most common serovars being S. 4,[5],12:i:-, a monophasic variant of Salmonella Typhimurium (36.6% of Salmonella-positive samples), followed by S. Derby (25.9% of Salmonella-positive samples). There was no significant difference compared to the estimate of overall prevalence (30.5% (95% CI 26.5–34.6)) obtained in the last abattoir survey conducted in the UK (2013). Abattoir-based control measures are often effective in the reduction of Salmonella contamination of carcasses entering the food chain. In this study, the effect of abattoir hygiene practices on the prevalence of Salmonella on carcasses was not assessed. Continuing Salmonella surveillance at slaughter is recommended to assess effect of farm-based and abattoir-based interventions and to monitor potential public health risk associated with consumption of Salmonella-contaminated pork products.

Technical specifications on a randomisation of sampling for the purpose of antimicrobial resistance monitoring from food-producing animals and food as from 2021.

To monitor antimicrobial resistance in zoonotic and indicator bacteria from food‐producing animal populations and meat thereof under Decision 2020/1729, a guidance for randomised sampling procedures is provided. Prospective and retrospective sampling plans for samples and isolates are addressed. The former involves collecting sufficient numbers of representative animal and food samples from which recovered isolates are tested for antimicrobial susceptibility; the latter involves selecting randomly Salmonella isolates from collections constituted within the framework of the national control programmes in poultry flocks. A generic proportionate stratified sampling process and numerical illustrations of proportional allocation are provided. Stratified sampling of Salmonella isolates from poultry primary productions is performed with proportional allocation to the size of the isolate collections available in official laboratories. An alternative approach would be a simple random sampling within the sampling frame of flocks positive for Salmonella. Stratified sampling of caecal samples, accounting for at least 60% of the domestic production of food‐producing animal populations monitored, with proportionate allocation to the slaughterhouse production, allows for the collection of representative isolates of Campylobacter and indicator E. coli and enterococci in various animal populations. Sampling of different chilled fresh meat categories is performed at retail outlets serving the final consumer, with proportional allocation of the number of samples to the population of geographical areas accounting for at least 80% of the national population, to test for the presence of ESBL‐/AmpC‐/carbapenemase‐producing E. coli. Stratified sampling of imported fresh meat is performed at border control posts, with proportional allocation to the number of consignments and origin to test Salmonella and indicator E. coli for antimicrobial susceptibility, and to test for the presence of ESBL‐/AmpC‐/carbapenemase‐producing E. coli. The corresponding sampling design is based on the reliable existing TRACES statistics, and the effect of the UK leaving the EU cannot be considered at this stage because of the major uncertainties still associated with it. These technical specifications should be updated as needed based on the first monitoring campaigns and trends in AMR.

Effects of Dietary Protein Levels on Bamei Pig Intestinal Colony Compositional Traits

Diets containing different crude protein levels (16%, 14%, and 12%) were created to feed Bamei pigs in order to study the effect of these compositions on intestinal colonies. Therefore, 27 healthy Bamei pigs of similar weight (20.99 kg ± 0.16 kg) were selected and randomly divided into three groups for microbial diversity analysis. The results of this study show that microbial diversities and abundances in Bamei pig jejunum and caecum samples after feeding with different dietary protein levels were significantly different. Dietary crude protein level exerted no significant effect on the Shannon index for cecum microbes in these pigs, while Simpson, ACE, and Chao1 indices for group I were all significantly higher than those of either the control group or group II (P < 0.05). Indeed, data show that microbial diversities and abundances in the 14% protein level group were higher than those in either the 16% or 12% groups. Dominant bacteria present in jejunum and cecum samples given low-protein diets were members of the phyla Firmicutes and Bacteroidetes. Data show that as dietary crude protein level decreases, representatives of the microbial flora genus Lactobacillus in jejunum and cecum samples gradually increases. Values for the KEGG functional prediction of microbial flora at different dietary protein levels also show that genes of jejunum and cecum microorganisms were mainly enriched in the “metabolism” pathway and indicate that low protein diets increase intestinal metabolic activity. Therefore, we recommend that Bamei pig dietary protein levels are reduced 2% from their existing level of 16% crude protein. We also suggest that essential synthetic amino acids (AA) are added to optimize this ideal protein model as this will increase intestinal flora diversity in these pigs and enhance health. These changes will have a positive effect in promoting the healthy growth of Bamei pigs.

Complementarity 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.

Variations of the Escherichia coli population in the digestive tract of broilers

ABSTRACT We explored the between-group and temporal variations in the intestinal Escherichia coli populations of broilers under experimental conditions, taking both antimicrobial resistance and virulence into consideration. Four replicates of 45 commercial chicks were reared in four animal facilities. On their first day of life (Day 0), they were orally inoculated with two extended-spectrum-cephalosporin-resistant (ESCR) E. coli (2.72 log10 CFU of a bla CMY-2- and 2.55 log10 CFU of a bla CTX-M-carrying E. coli). Faecal samples were then collected weekly and caecal samples were obtained from birds sacrificed on Days 21 or 42. The total, ESC-, ciprofloxacin- and gentamicin-resistant E. coli populations were enumerated on MacConkey (MC) and MC-supplemented media, and eight virulence-associated genes (VAGs) (iroN, iutA, iss, ompT, hlyF, vat, frzorf4 , and fyuA) were sought by PCR on isolates obtained on MC agar. The results showed significant between-group differences in the size of the resistant sub-populations and the presence of VAGs. Contrary to bla CTX-M-positive strains, bla CMY-positive strains persisted up to Day 42, but represented only a minor fraction of the total E. coli population. The ESC-, gentamicin- and ciprofloxacin-resistant populations decreased over time. Isolates obtained during the first week contained a mean of 5.1 VAGs. The percentages of some VAG profiles differed between faecal isolates on Day 41 and caecal isolates on Day 42. The fluctuations or differences between E. coli isolates according to group, age, and faecal or caecal origin need to be considered when designing experimental protocols and seeking to improve colibacillosis control. RESEARCH HIGHLIGHTS Temporal variations in the intestinal E. coli populations of broilers was studied. The antibiotic-resistant populations decreased over time. Virulence profiles differed between faecal isolates on Day 41 and caecal isolates on Day 42. Strains with the highest numbers of virulence genes were present during the first days.

Assessing Campylobacter Colonization of Broiler Herds Ante Mortem and Monitoring Campylobacter Contamination Post Mortem by qPCR.

Human campylobacteriosis is the most prevalent zoonosis, with chicken meat contributing substantially to the number of cases. Measures to avoid or at least reduce exposure by meat contaminated with Campylobacter (C.) spp. are needed. With regard to the process hygiene criterion introduced in 2018 for Campylobacter spp. on broiler carcasses, we evaluated the performance of a recently developed quantitative real-time PCR (qPCR) for C. jejuni/coli on random caecal samples and chicken meat. With the qPCR on pooled caecal samples not only C. jejuni/coli positive (69.6%) versus negative broiler herds (30.4%) were identified, but herds highly colonized with C. jejuni/coli (39.4%) could also be identified. From the chicken meat samples, 8.0% were positive for C. jejuni/coli by qPCR and 0.7% by enumeration (>10 cfu/g) compared to 58.3% using cultural enrichment. Given the higher sensitivity, the qPCR method could replace the currently used enumeration method to assess the process hygiene criterion for Campylobacter spp. on broiler carcasses. Moreover, with the qPCR, a reliable identification of C. jejuni/coli colonized incoming broiler herds a few days before slaughter is feasible, which provides important information to optimize slaughter processes. Finally, identifying and monitoring herds with high C. jejuni/coli colonization rates could help to individually improve biosecurity measures at farm level, eventually reducing the C. jejuni/coli load on chicken meat.

Inhibitory Effect of Herbs on Extended Spectrum Beta Lactamase Enzyme from Escherichia coli of Healthy Broilers

Background: Bacterial resistance to beta-lactam antibiotics has risen dramatically in Escherichia coli from poultry and other food animals due to the production of extended-spectrum beta-lactamase enzyme (ESBLs) which degrades third generation cephalosporins.Herbs could be a better alternative for such cases. Methods: Present study was undertaken on 400 caecal samples of healthy broilers, collected from various poultry sale outlets of Jabalpur (M.P.). Samples were screened phenotypically and genotypically for the presence of extended spectrum beta lactamase producing E. coli. Inhibitory effect of fruit peel juice of Punica granatum and Syzigium aromaticum oil observed on the Beta lactamase enzyme obtained from these positive samples by colorimetric method using CENTA and NITROCEFIN as chromogenic substrate.Result: Phenotypically 135 samples/isolates were ESBL producing E.coli. On multiplex PCR assay 76 positive samples with bla TEM, blaCTX and bla SHV, gene was obtained. Oil of Syzygium aromaticum showed maximium per cent inhibition, Punica granatum depicted lower per cent inhibition with CENTA and NITROCEFIN respectively. Combination of both herbs showed increased inhibition. Tazobactum, (100µM) taken as the standard control exhibited 99.88 and 98 per cent inhibition of ESBL enzyme.

Growth performance, gut health, carcass yield traits and profitability of broiler chicken raised on compound diet supplemented with probiotics

The present study was conducted to investigate the effects of probiotic on growth performance, gut health, viability, carcass yield traits and cost benefit analyses etc., of broiler chicken from d1-28 days. A total of 192 day-old broiler chicks (Ross 308) was assigned into four dietary treatment groups, i.e. D1 (Control), D2 (Poultry Starsol), D3 (Avilac plus) and D4 (Avibac), and each treatment replicated six times with eight birds per replicate in a completely randomized design. The chicks were raised in battery cages all the trial period. Ready-made starter diet (crumble) was fed the chicks up to 14 days, after that, grower (pellet) diet fed the broiler from d15 to 28 days. The water treated with probiotics at the rate of 1.0 g L-1 (Poultry Starsol), 1.0 ml L-1 (Avilac plus) and 1.0 g L-1 (Avibac) in D2, D3 and D4 treatment groups, respectively, and supplied the birds ad libitum entire the trial period. Data on feed intake (FI), body weight gain (BWG), feed conversion ratio (FCR) and livability (%) were collected. Apart from this, caecal digesta sample was taken on 28 days to determine the gut microflora population, i.e. total viable count (TVC) and total lactobacilli count (TLC) of broilers. Carcass characteristics (dressing %, blood weight, drumstick weight, thigh weight, breast weight %) and cost benefit were also calculated on the last day of trial period to assess the different meat cuts and the profitability of broiler. The data revealed that FI, viability (%), TVC and TLC of broilers had no difference (P&gt;0.05) between treatment on 28d. Except for first week, BWG was improved significantly (P&lt;0.05) in the birds fed probiotics during d1-21 and d1-28, respectively. Superior FCR values (1.30, 1.34) were observed in the broilers of probiotics supplemented groups from d1-28 days of age. The results of carcass traits were influenced significantly (P&lt;0.05) by dietary treatment. The percentage of breast weight, thigh weight, dressing yield, drumstick weight and blood weight was increased (P&lt;0.05) in the supplemental group compared to control. Higher (P&lt;0.01) net profit was observed in the birds of probiotics supplemented group than that of control group. It can be concluded that broiler responded positively as a result of probiotics supplementation in water, and can be raised profitably under farming condition with low cost.&#x0D; Int. J. Agril. Res. Innov. Tech. 10(1): 28-34, June 2020

MORPHOLOGY OF THE CAECUM IN LABORATORY ANIMALS IN HEALTH: SPECIES COMPARISON

In recent decades, the prevalence of gastrointestinal diseases has increased thus posing the immediate, both therapeutic and surgical treatment. It brings forth a problem of searching new and improving existing approaches and techniques for correcting the above-mentioned diseases. Preclinical studies in this area are conducted exclusively on laboratory animals and peculiarities of the morphological features of their organs are of great importance when comparing with the human morphology. The methodology used in the study included histological, morphometric and statistical techniques; biopsy samples of caecum taken from 5 rabbits were investigated. We assessed the correctness of the trait distribution by each of the variations, the mean values for each trait studied, standard errors and standard deviations. The significance of the difference of values between independent micrometric values in the normal trait distribution was determined by Student's criterion. The paper describes the main morphological characteristics of the caecum in rabbits and compared the findings obtained with similar structures of the human caecum. The caecum of rabbits, as of humans, has four layers: mucous, submucosal, muscular and serous. The mucous membrane consists of the epithelial layer located on the basement membrane and the muscular plate and contains cellular elements. The submucosa is composed of loose fibrous connective tissue, which contains collagen and reticular fibres, elements of diffuse lymphoid tissue, blood vessels, and nerve endings. The muscular and serous membranes are quite similar to the human caecum. Thus, the optic light microscopy has demonstrated the morphology of the caecum in rabbits is similar to that in the human caecum.

Effects of Ligustrum lucidum on egg production, egg quality, and caecal microbiota of hens during the late laying period

This study was conducted to determine the effects of diet supplementation of laying hens with Ligustrum lucidum (LL) on egg production, egg quality and caecal microbiota. A total of 360 72-week-old Hy-Line Brown laying hens were divided into three groups with four replicates of 30 birds each. The laying hens were fed with basal diet (control group), basal diet + 1% LL (group I) and basal diet + 2% LL (group II). The experiment lasted for 45 d. Eggs were collected daily and caecal samples were collected at the end of the experiment. Results showed that dietary supplementation with LL did not affect the average daily egg weight, the average daily feed intake, the cracked egg rate, the mortality and the egg quality (p>.05). However, groups I and II showed significantly increased laying rate and decreased feed/egg ratio (p＜.05). The differences in caecal microbiota between group II and the control group were significant. The relative abundance of Bacteroidetes, Firmicutes, Saccharibacteria and Verrucomicrobia at the phylum level; Ruminococcaceae, Rikenellaceae, Acidaminococcaceae, unclassified_Bacteroidales, S24-7 group and ODP1230B8.23 at the family level; RC9 gut group, Phascolarctobacterium, unclassified_Bacteroidales, Butyricicoccus, norank_f__Ruminococcaceae, Ruminococcaceae_UCG-014, coprostanoligenes group and Ruminococcaceae_UCG-005 at the genus level in group II changed significantly compared with that in the control group (p＜.05). Dietary supplementation with 1% and 2% LL could improve the laying performance and affect the caecal microbial community structure of laying hens during the late laying period. Highlights Dietary supplementations with Ligustrum lucidum significantly increased laying performance of hens during the late laying period. Dietary supplementations with Ligustrum lucidum not affected the egg quality of laying hens. Dietary supplementations with Ligustrum lucidum affected the microbial structure of caecum of laying hens.

Entecavir therapy reverses gut microbiota dysbiosis induced by hepatitis B virus infection in a mouse model

IntroductionChronic hepatitis B virus (HBV) infection is a global public health problem. The gut microbiota has been linked to pathogenesis of liver diseases induced by chronic HBV infection. Methods and ResultsThis study established a recombinant adeno-associated virus serotype 8 (rAAV8)-mediated persistent HBV infection mouse model. Entecavir (ETV) treatment significantly decreased the HBV DNA load both in serum and the liver. The comparison of gut microbiota composition of rAAV8-HBV-infected mice and ETV-treated mice with healthy controls was carried out using 16S rDNA sequencing analysis of caecal content samples. The intestinal microbiota alpha diversity of rAAV8-HBV-infected mice decreased, and significantly restored after 4 weeks of ETV therapy. Blautia and Clostridium sensu stricto significantly decreased in rAAV8-HBV-infected mice and was negatively correlated with both HBsAg and HBeAg levels. On the contrary, the Butyricicoccus and Prevotellaceae NK3B31 groups exhibited positive correlation with HBsAg and HBeAg. Furthermore, it was observed that Akkermansia, a known gut barrier-protecting bacterium, significantly decreased in rAAV8-HBV-infected mice and was restored to the level of that in healthy controls after ETV therapy, while the abundance of Akkermansia was negatively correlated with HBV DNA load both in serum and the liver. ConclusionTaken together, the results showed that dysbiosis of gut microbiota developed in the persistent HBV-infected mice and was effectively reversed by ETV treatment, shedding light on the mechanisms of gut microbiota on HBV-persistent infection and antiviral therapy.

Biofilm forming properties of quinolone resistant Escherichia coli from the broiler production chain and their dynamics in mixed biofilms

BackgroundQuinolone resistant Escherichia coli (QREC) have been found in samples from Norwegian broiler chicken, despite quinolones not being administered to poultry in Norway. Biofilm production may be one factor contributing to the observed persistence in the broiler production chain. In the present study, 158 QREC strains from chicken caecal and retail meat samples were screened for biofilm production in microtiter plates, biofilm morphotype on Congo Red (CR) agar plates and phylotype by multiplex PCR. Furthermore, the dynamics in mixed biofilms with strains of different morphotypes were studied on glass slides and on CR agar plates.ResultsAll strains but one produced biofilm in microtiter plates and/or on CR agar plates at room temperature. There were no differences between strains from chicken caecum and chicken retail meat in the mean amount of biofilm produced in microtiter plates. Furthermore, no differences in biofilm production were observed between phylotypes. However, significant differences in biofilm production were found between biofilm morphotypes. The morphotype RDAR (red dry and rough, which has both curli and cellulose in the matrix, was displayed by 70% of the strains. Mean biofilm production by these strains were significantly higher than by strains with the morphotypes PDAR (pink dry and rough) with only cellulose or BDAR (brown dry and rough) with only curli. Interestingly, the two latter morphotypes produced biofilms with the morphotype RDAR when grown together. None of the strains achieved significantly higher numbers of colony forming units (cfu) in mixed biofilms than in single strain biofilms on glass slides.ConclusionsThe results indicate that QREC can form biofilm reservoirs on both inert and organic surfaces in production environments, as well as on meat. This may contribute to persistence and dissemination of the strains. Strains with both curli and cellulose in the biofilm matrix were significantly better biofilm formers than strains lacking one of these components. However, strains with only one of the components could compensate for this by producing mixed biofilms with strains having the other component, and thereby most likely enhance their probabilities of persistence in the production environment.

Immuno- and glyco-histochemistry as a tool to evaluate the oregano supplemented feed effects in pig gut.

Among oregano properties, its antioxidant and antibacterial effects are particularly interesting. Oregano is also able to induce a higher glycoconjugate production in gut, creating a physical barrier against microorganisms. This study evaluated the effects of adding an aqueous extract of oregano (OAE) to the diet of two homogenous groups of pigs during the finisher phase. The diets were as follows: control commercial diet (CTR group) and CTR diet supplemented (2 g/kg) with OAE (O group). Samples of ileum and caecum from the two groups were examined by conventional histochemistry to analyze complex carbohydrates and by immunohistochemistry to detect Bcl-2 Associate X protein (BAX), an indicator of oxidative stress. Glyco-histochemistry showed significant differences between the two groups. Immunohistochemistry revealed a lower presence of BAX in O group. The OAE supplementation improved the production of glycoconjugates, able to enhance in pig the protection of intestinal mucosa by means of direct and indirect defense actions. The reduced BAX immunostaining observed in O group may be an indicator of enhanced antioxidant action promoted by oregano. The results of this study can be used in further research to identify ways to improve endogenous defence ability, with the aim of reducing antibiotic use and preventing antimicrobial resistance.

Assembly of hundreds of novel bacterial genomes from the chicken caecum

BackgroundChickens are a highly important source of protein for a large proportion of the human population. The caecal microbiota plays a crucial role in chicken nutrition through the production of short-chain fatty acids, nitrogen recycling, and amino acid production. In this study, we sequence DNA from caecal content samples taken from 24 chickens belonging to either a fast or a slower growing breed consuming either a vegetable-only diet or a diet containing fish meal.ResultsWe utilise 1.6 T of Illumina data to construct 469 draft metagenome-assembled bacterial genomes, including 460 novel strains, 283 novel species, and 42 novel genera. We compare our genomes to data from 9 European Union countries and show that these genomes are abundant within European chicken flocks. We also compare the abundance of our genomes, and the carbohydrate active enzymes they produce, between our chicken groups and demonstrate that there are both breed- and diet-specific microbiomes, as well as an overlapping core microbiome.ConclusionsThis data will form the basis for future studies examining the composition and function of the chicken caecal microbiota.

Prevalence and Antibiotic Resistance Pattern of Salmonella Isolated from Caecal Contents of Exotic Chicken in Debre Zeit and Modjo, Ethiopia.

A cross-sectional study was conducted between December, 2013, and May, 2014, to determine the prevalence and antibiotic resistance feature of Salmonella isolated from broilers slaughtered in Debre Zeit and Modjo towns, Ethiopia. A total of 384 caecal content samples were collected for microbiological examination following the standard techniques and procedures outlined by the International Organization for Standardization to isolate Salmonella. The sensitivity of the isolates subjected to nine antimicrobials was tested by the Kirby–Bauer disk diffusion method. The overall prevalence of Salmonella was 14.6%, and its occurrence differ significantly by farm (p < 0.05). The occurrence of the bacteria was not statistically different in the midland (15.2%) and lowland (13.3%) (p > 0.05) and between males (13.5%) and females (15.6) (p > 0.05). Of the 50 isolates, 48 were resistant to at least one drug. Multidrug resistance was recorded in 43 (86.0%) of the isolates. The study demonstrated considerable prevalence and high antimicrobial resistant Salmonella in exotic chicken and indicates the potential importance of chickens as source of foodborne salmonellosis and multiple antimicrobial resistance of Salmonella. Improving the hygienic practice of farms could help to reduce the occurrence of Salmonella in farms. Further studies are needed to describe the risk factors associated with the emergence of drug-resistant Salmonella in chicken.

Prevalence and characteristics of Livestock-Associated Methicillin-Resistant Staphylococcus aureus (LA-MRSA) isolated from chicken meat in the province of Quebec, Canada

This study was conducted to estimate the prevalence of Livestock-Associated Methicillin-Resistant Staphylococcus aureus (LA-MRSA) in retail chicken meat and broiler chickens from the Province of Quebec, Canada, and to characterize LA-MRSA isolates. A total of 309 chicken drumsticks and thighs were randomly selected in 2013 from 43 retail stores in the Monteregie. In addition, nasal swabs and caeca samples were collected in 2013–2014 from 200 broiler chickens of 38 different flocks. LA-MRSA was not detected in broiler chickens. Fifteen LA-MRSA isolates were recovered from four (1.3%) of the 309 chicken meat samples. Multi-Locus Sequence Typing (MLST) and SCCmec typing revealed two profiles (ST398-MRSA-V and ST8-MRSA-IVa), which were distinct using pulse-field gel electrophoresis (PFGE) and microarray (antimicrobial resistance and virulence genes) analyses. In addition to beta-lactam resistance, tetracycline and spectinomycin resistance was detected in all isolates from the 3 positive samples of the ST398 profile. Southern blot hybridization revealed that the resistance genes aad(D) and lnu(A), encoding resistances to aminoglycosides and lincosamides respectively, were located on plasmid. All isolates were able to produce biofilms, but biofilm production was not correlated with hld gene expression. Our results show the presence of two separate lineages of MRSA in retail chicken meat in Quebec, one of which is likely of human origin.

Effects of Antimicrobial peptides on egg production, egg quality and caecal microbiota of hens during the late laying period.

This study was conducted to determine the effects of diet supplementation of laying hens with antimicrobial peptides (AMP) on egg production, egg quality and caecal microbiota. A total of 360 Hy-Line Brown laying hens (72weeks old) were divided into three groups with four replicates of 30 birds each. The laying hens were fed with the basal diet (Control), the basal diet+50mg/kg AMP (group 1) and the basal diet+100mg/kg AMP (group 2). The experiment lasted for 45 d. Eggs were collected daily and caecal samples were collected at the end of the experiment. The results showed that AMP supplementation caused a significantly increased laying rate and decreased feed/egg ratio (p＜.05). Meanwhile, a distinctive difference in cecal microbiota was observed between AMP and control groups and the average values of microbial diversity and richness were lower in the AMP group than in the control group. At the phylum level, the relative abundance of Verrucomicrobia and Cyanobacteria were lower in the AMP group than in the control group. In conclusion, the results indicated that dietary supplementation with AMP can improve egg production and affect the cecal microbial community membership and structure of hens during late laying period.

Digestive Tract of Beavers (Castor fiber) Associated with Staphylococcal Species Variability and Their Properties

Objective: Staphylococci from beavers‘ gut (Castor fiber) were analysed as a contribution in the basic microbiology as well as in the part of beavers‘ microbiome study. Methodology: Free-living beavers (12), both male and female (aged 4-5 years) were caught with a net in north-east part of Poland Województwo (Provincie) Podlaskie Gmina-Wizajny, GPS:22º 52E:54º22N and placed in wire cages. Sampling was provided with the Polish colleagues. Samples of jejunum (12), colon (12) and caecum (6) were done according to all ethic rules for animal handling. Results: Staphylococci detected in jejunum reached 2.73 ± 1.16 cfu/g (log 10) on average; their counts in caecum reached 1.87 ± 0.37 cfu/g on average and in colon 2.89 ± 1.70 cfu/g. After score evaluation using MALDI-TOF spectrometry, a high variability in staphylococcal strains/species distribution in beavers‘ gut was found; in total, including 18 identified strains, nine species were detected belonging into five clusters; all strains were allotted in the coagulase-negative staphylococcal species. The most frequently detected species were S. hominis and S. haemolyticus (five strains for each). The other species included two strains-S. epidermidis and S. lentus, S. pasteuri, S. cohnii, S. vitulinus, S. warneri, S. xylosus, one strain for each one species. Fifteen strains were DNase and almost hemolysis negative. Ten strains (not depending on the species) showed low-grade biofilm ability. Most of strains were methicillin susceptible with high production of lactic acid. Conclusion: The study is original contribution in staphylococcal microbiome of beavers and gives opportunity in more detail study of individual strain species.