Chicken Feces Research Articles

Molecular detection of virulence genes in Salmonella spp. isolated from chicken faeces in Mafikeng, South Africa.

Chickens have been implicated in most Salmonella disease outbreaks because they act as carriers of the pathogen in their gut. There are over 2500 serotypes of Salmonella that have been reported worldwide and 2000 of these serovars can be found in chickens. The main objective of this study was to determine the Salmonella serotypes found in poultry farms around Mafikeng district, South Africa. Salmonella was identified according to the guidelines of the International Organization for Standardization (ISO) (ISO 6579:2002) standard techniques. Faecal samples were collected and analysed for Salmonella using conventional cultural methods and polymerase chain reaction targeting the 16S Ribosomal Deoxyribonucleic acid (rDNA) gene for Salmonella identification. Out of 130 presumptive Salmonella isolates determined by urease and triple sugar iron tests, only 46 isolates were identified as Salmonella serotypes of which S. Typhimurium was the most frequent with 18 (39.1%), followed by S. Heidelberg with 9 (19.6%), S. bongori with 7 (15.2%), S. Enteritidis with 6 (13.0%) and both S. Paratyphi B and S. Newport with 3 (6.5%) each. Seven virulence genes including invA 100%, spy 39%, hilA 9%, misL 30%, sdfI 13%, orfL 11% and spiC 9% were detected from these Salmonella isolates in this study. The presence of these virulence genes indicates high pathogenicity potential of these isolates which is a serious public health concern because of zoonotic potential of Salmonella.

High prevalence of mcr-1 encoding colistin resistance and first identification of blaCTX-M-55 in ESBL/CMY-2-producing Escherichia coli isolated from chicken faeces and retail meat in Tunisia

Avian industries have been reported as an important contributor in the worldwide spread of antibiotic resistance owing to some particular practices especially the overuse of antibiotics. Thus in this study, we aimed to characterize extended-spectrum-beta-lactamase (ESBL) and acquired-AmpC-beta-lactamase (aAmpC)-producing Escherichia coli isolates from chicken faeces and raw meat in Tunisia. During the year 2018, 286 faecal chicken swabs and 47 raw chicken meat samples were collected and processed to recover cefotaxime-resistant E. coli. Antimicrobial susceptibility was performed by disk-diffusion and/or broth-microdilution. blaTEM, blaSHV, blaCTX-M, and blaCMY genes were investigated by PCR/sequencing. Genes encoding resistance to colistin (mcr-1 to mcr-8), tetracycline (tetA/tetB), sulfonamide (sul1/sul3), and chloramphenicol (cmlA), were analysed by PCR. Class 1 integrons were investigated by PCR/sequencing. Phylogenetic groups of all isolates were determined. PFGE and MLST were performed for representative isolates. PCR-based replicon typing was performed in mcr1-harbouring isolates. Cefotaxime-resistant E. coli was detected in 22.4% (64/286) and 63.8% (30/47) of faeces and meat samples, respectively. Ninety isolates were ESBL-producers and harboured the genes: blaCTX-M-1 +/− blaTEM-1 (n = 65), blaCTX-M-55 +/− blaTEM-1 (n = 21), blaCTX-M-14 (n = 1), and blaSHV-12 (n = 3). The blaCMY-2 gene was detected in four ESBL-negative isolates. Isolates belonged to phylogroups D (50%), A (36.2%), B1 (9.6%), and B2 (4.3%). Fifty-four were colistin-resistant and 52 carried the mcr-1 gene. The tetA, sul1/sul3 and cmlA genes were detected among resistant isolates and 76 harboured class 1 integrons. MLST analysis revealed 13 sequence types (STs). The isolates were classified into 28 PFGE types. The IncP, IncFIB, and IncI1 replicons were detected among mcr-1-positive strains. We report a high frequency of ESBL-producers and colistin-resistant E. coli in chicken and derived food and the detection for the first time of blaCTX-M-55 in poultry in Tunisia.

Effect of Storage Temperature on the Survival of New Zealand Egg-Associated Salmonella Isolates in and on Eggs

The influence of egg storage temperature on Salmonella contamination of eggs is a key consideration in determining storage and shelf life recommendations for eggs. In this study, experiments assessed the survival of Salmonella isolates on and in eggs at storage temperatures (15 and 22°C) currently used in New Zealand. Eggshell surfaces were inoculated with a cocktail of 10 Salmonella isolates comprising five serotypes, at a concentration of ∼106 CFU per egg (for determining shell surface survival) or ∼103 CFU per egg (for determining internalization). Additionally, a subset of eggs was artificially contaminated with sterile chicken feces prior to Salmonella inoculation. Inoculated eggs were incubated at 15 and 22°C. At 0, 21, and 35 days of incubation, eggshells were enumerated for Salmonella, and egg contents were tested for Salmonella presence or absence (yolk) or most probable number (albumen). Higher levels of Salmonella were recovered from eggshells following incubation at 15°C (31% relative humidity [RH]) compared with 22°C (45% RH) after both 21 and 35 days of incubation. Recoverable numbers of Salmonella from visibly clean eggshell surfaces declined over time at both storage temperatures and were at, or below, the limit of detection from eggs stored at 22°C and 45% RH for 35 days. A substantially higher concentration of viable Salmonella was recovered from eggshells that were experimentally contaminated with chicken feces compared with those without, particularly from eggs stored at 15°C and 31% RH for 35 days (2.38 log higher CFU from eggs containing feces). No Salmonella was detected in egg contents (albumen or yolk) at any incubation temperature or time point, regardless of the presence of feces. Findings emphasize the importance of current regulations that require eggs sold at retail to be visibly clean and will inform risk management decisions regarding egg storage times and temperatures with respect to Salmonella control in and on New Zealand eggs at retail.

Limited changes in the fecal microbiome composition of laying hens after oral inoculation with wild duck feces

Interspecies transmission of fecal microbiota can serve as an indicator for (indirect) contact between domestic and wild animals to assess risks of pathogen transmission, e.g., avian influenza. Here, we investigated whether oral inoculation of laying hens with feces of wild ducks (mallards, Anas platyrhynchos) resulted in a hen fecal microbiome that was detectably altered on community parameters or relative abundances of individual genera. To distinguish between effects of the duck inoculum and effects of the inoculation procedure, we compared the fecal microbiomes of adult laying hens resulting from 3 treatments: inoculation with wild duck feces (duck), inoculation with chicken feces (auto), and a negative control group with no treatment. We collected cloacal swabs from 7 hens per treatment before (day 0), and 2 and 7 D after inoculation, and performed 16S rRNA amplicon sequencing. No distinguishable effect of inoculation with duck feces on microbiome community (alpha and beta diversity) was found compared to auto or control treatments. At the individual taxonomic level, the relative abundance of the genus Alistipes (phylum Bacteroidetes) was significantly higher in the inoculated treatments (auto and duck) compared to the control 2 D after inoculation. Seven days after inoculation, the relative abundance of Alistipes had increased in the control and no effect was found anymore across treatments. These effects might be explained by the perturbation of the hen's microbiome caused by the inoculation procedure itself, or by intrinsic temporal variation in the hen's microbiome. This experiment shows that a single inoculation of fecal microbiota from duck feces to laying hens did not cause a measurable alteration of the gut microbiome community. Furthermore, the temporary change in relative abundance forAlistipes could not be attributed to the duck feces inoculation. These outcomes suggest that the fecal microbiome of adult laying hens may not be a useful indicator for detection of single oral exposure to wild duck feces.

Characterization and comparison of yeasts from different sources for some probiotic properties and exopolysaccharide production

Abstract A total of 134 strains of yeasts isolated from fruits and vegetables (certain berry fruits, apples, pomegranates, carrots and grapes), free-range chicken feces and dairy products, and from 10 commercial yeast preparations were identified and subjected to analyses to determine their in vitro probiotic properties. Based on 26S rRNA sequence analysis all ten isolates from commercial products were identified as Saccharomyces cerevisiae, and the natural isolates as Candida corpophila, Candida diddensiae, Clavispora lusitaniae, Hanseniaspora opuntiae, Hanseniaspora uvarum, Kazachstania bovina, Kluyveromyces marxianus, Metschnikowia. pulcherrima, Metschnikowia sp., Meyerozyma carribbica, Pichia kluyveri and Wickerhamomyces anomalus. All isolates were found to be resistant to simulated gastric juice at pH 2.5 for 2 h and were able to grow at both 30 and 37 °C. The exopolysaccharide (EPS) production of isolates from commercial preparations and from natural sources varied between 249-275.22 and 27.95-272.22 mg/L, respectively. Two of the natural isolates had levels of EPS production comparable to the natural strains (S. cerevisiae T8-3C and S. cerevisiae P25-1) with 264.63 and 272.53 mg/L, respectively. Isolates were also investigated for autoaggregation and coaggregation abilities. The highest coaggregation ability was determined for the Saccharomyces cerevisiae P25-1 strain against Staphylococcus aureus (ATCC 25923).

The Prevalence of Gastrointestinal Helminths in Domestically Farmed Chicken in Van, Turkey

The present study was conducted with the aim to determine the prevalence of gastrointestinal helminth in domestically farmed chicken in İpekyolu, Tuşba and Edremit districts in Van Province in Turkey. Feces of 185 chicken were examined and 78 chicken (42.16%) yielded positive results for single species, 26 (14.05%) were positive for two species, and 6 chicken (3.25%) were positive for three species. Six helminth eggs, four nematodes and two cestodes, were detected in infected chicken feces. Those infected with single species were studied based on helminth and infection types and the rates of infection was determined as: Capillaria spp. 23 (12.50%), Heterakis gallinarum 19 (10.32%), Ascaridia spp. 18 (9.78%), Raillietina spp. 8 (4.35%), Davania proglottina 6 (3.26%), Trichostrongylus tenuis 4 (2.17%). The infection determined in feces caused by two species were Capillaria spp. + Heterakis gallinarum 10 (5.43%), Capillaria spp. + Ascaridia spp. 8 (4.35%), Heterakis gallinarum + Ascaridia spp. 7 (3.78%), Raillietina spp. + Davania proglottina 1 (0.54%) and infection determined in feces caused by three species were determined as Capillaria spp. + Heterakis gallinarum + Davania proglottina 4 (2.17%) and Capillaria spp. + Heterakis gallinarum + Trichostrongylus tenuis 2 (1.09%). The findings of the present study indicated that helminth infections were significantly prevalent in domestic chicken in Van Province in Turkey.

Cash for trash: an agro-economic value assessment of urban organic materials used as fertilizers in Cameroon

The rapid expansion of cities in sub-Saharan Africa generates increasing volumes of diversified organic wastes that require management and a high population need for local and fresh vegetables. Linking the different sources of organic materials generated in these cities with the needs of local food producers for fertilizers prepared from recycled organic materials is challenging. This requires that producers obtain the results they expect from the organic materials they recycle. Thus, recycling these organic materials in their raw form or after composting presents an opportunity for periurban agriculture, although the great variability in the quantity and quality of these materials raises questions about the consistency between their agronomic value and their economic value. In two large cities of Cameroon (Yaoundé and Bafoussam), different types of organic materials were sampled: unprocessed livestock waste and composts from manure or municipal solid waste. From an agronomic perspective, we calculated the fertilization value based on the nutrient (nitrogen, phosphorus, and potassium) content of the organic materials. From an economic perspective, we calculated the value based on the nutrient content of the organic materials and their substitution unit prices and recorded the current market price. This is the first time that this combined approach has been used in sub-Saharan Africa. Our results showed considerable variability and discrepancy in both the agronomic and the economic values. The market prices overvalued the urban composts by a factor of 6, while chicken feces were undervalued by a factor of 3. The unprocessed organic materials were the most interesting from an economic and agronomic perspective. Our findings suggest that (i) the composting process needs to be improved and (ii) the humus potential should be calculated to better assess the amendment value of organic materials and as a basis for adjusting their market price.

Genome Analysis and Multiplex PCR Method for the Molecular Detection of Coresistance to Cephalosporins and Fosfomycin in Salmonella enterica Serovar Heidelberg

Heidelberg is among the top three Salmonella enterica serovars associated with human foodborne illness in Canada. Traditional culture and antimicrobial susceptibility testing techniques can be time-consuming to identify Salmonella Heidelberg resistant to cephalosporins and fosfomycin. Rapid and accurate detection of such antibiotic-resistant Salmonella Heidelberg isolates is essential to adopt appropriate control measures. In this study, 15 Salmonella Heidelberg strains isolated from feces of Canadian broiler chickens were characterized by whole genome sequencing. Salmonella Heidelberg genomes had an average coverage of greater than 80-fold, an average of 4,761 protein-coding genes, and all belonged to multilocus sequence type ST15. Genome sequences were compared with genomes in the National Center for Biotechnology Information Pathogen Detection database ( www.ncbi.nlm.nih.gov/pathogens/ ), including human outbreak isolates. The Canadian broiler isolates clustered with chicken isolates from the United States and an equine clinical isolate from Ontario, Canada. In agreement with their antimicrobial resistance phenotypes, several chromosomally encoded specific antimicrobial resistance genes including fosA7 and multidrug resistance efflux pump determinants were detected. An AmpC-like β-lactamase gene, blaCMY-2, linked with a quaternary ammonium compound resistance gene, sugE, on a replicon type IncI1 plasmid was detected in all 15 broiler Salmonella Heidelberg isolates. Of the 205,031 published Salmonella genomes screened in silico, 4,954 (2.4%) contained blaCMY-2, 8,143 (4.0%) contained fosA7, and 919 (0.4%) contained both resistance genes. The combination of both resistance genes (fosA7 and blaCMY-2) was detected in 64% of the Heidelberg genomes and in a small proportion of various other serovars. A PCR method was developed to detect Salmonella Heidelberg in pure culture and chicken feces based on specific primers targeting genes conferring fosfomycin (fosA7) and third-generation cephalosporin (blaCMY-2) resistance as well as the Salmonella-specific invA gene and the universal 16S rRNA genes. The PCR assay was specific and sensitive for blaCMY-2 and fosA7 harboring Salmonella Heidelberg. However, some other Salmonella serovars containing these two resistance genes could also be detected by the developed PCR method.

The Yersinia high-pathogenicity island (HPI) carried by a new integrative and conjugative element (ICE) in a multidrug-resistant and hypervirulent Klebsiella pneumoniae strain SCsl1

Multidrug-resistant and hypervirulent Klebsiella pneumoniae (hvKP) poses a significant risk to public health. To better understand the molecular characteristics of multidrug-resistant and hypervirulent K. pneumoniae of animal origin, fifteen K. pneumoniae strains from the liver, blood of sick pigs and chicken feces were collected. All K. pneumoniae isolates were subjected to antimicrobial susceptibility testing, string test, multi-locus sequence typing and whole genome sequencing. Seven K. pneumoniae isolates were found carrying the mcr-1.1 gene. Among them, a multidrug-resistant and hypervirulent K. pneumoniae strain SCsl1 isolated from the liver of a diseased pig was found to harbor 16 resistance genes (e.g., mcr-1.1) and 16 virulence genes including aerobactin. Moreover, a novel integrative and conjugative element, named ICEKpSL1, was identified in SCsl1, which contains a full Yersinia high-pathogenicity island (HPI). This element could be excised from the chromosome to form a circular intermediate, indicating potential transmission of the Yersinia pathogenicity island. The emergence of multidrug-resistance and hypervirulence in K. pneumoniae from animals warrants further surveillance.

A Sensitive, Specific and Simple Loop Mediated Isothermal Amplification Method for Rapid Detection of Campylobacter spp. in Broiler Production

Campylobacteriosis is one of the most common foodborne diseases worldwide. Two Campylobacter species – C. jejuni and C. coli in poultry and poultry products are considered to be the main source of human campylobacteriosis. Therefore, studying Campylobacter status in poultry flocks is needed to prevent transmission of disease and reduce human risk, health cost, and economic losses. In this study, we adapted and used a Loop-Mediated Isothermal Amplification (LAMP) assay for specific, sensitive, simple and cost-effective rapid detection of C. jejuni and C. coli in the poultry production chain. Amplified LAMP products were detected using a small, low-cost portable commercial blue LED transilluminator and a direct visual detection strategy was demonstrated. By using optimized conditions for amplification a limit of detection (LOD) of 50 CFU/ml was achieved for testing of C. jejuni and C. coli in spiked chicken feces without enrichment. The method took 60–70 min from receiving the samples to the final results (including 30 min for amplification). The optimized LAMP showed a relative accuracy of 98.4%, a specificity of 97.9%, and a sensitivity of 100% in comparison to real-time PCR method. Cohen’s kappa index also showed an excellent agreement (0.94) between the two methods. The results showed that the method is specific, sensitive and is suitable to develop for rapid detection of Campylobacter spp. at poultry production.

An efficient method for extracting microplastics from feces of different species

Concerns have been raised regarding the ingestion of microplastics (MPs) by numerous organisms including humans. However, no efficient and standardized methods are available for extracting MPs from feces. In this study, we introduce a novel approach with high digestion efficiency that involves using Fenton’s reagent and nitric acid to remove feces solids. Firstly, Fenton’s reagent was used to degrade small solids and decompose large solids into small pieces. Secondly, nitric acid was used to digest the remaining solids and filters. Furthermore, absolute ethyl alcohol was used to remove the mineral residues wrapped on the plastic surfaces and disperse MPs. By using this method, 97.78 % MPs can be recovered from human and chicken feces, and no significant changes were observed in the physical and Raman spectral properties of different polymer types of MPs. This method has also been verified by extracting MPs from field feces. Overall, the proposed method can efficiently digest feces solids and extract MPs with higher recovery rate, less intermediate steps and less damage, which can serve as an economical and feasible method for the detection of MPs in the feces of different species.

From farm to fork: identical clones and Tn6674-like elements in linezolid-resistant Enterococcus faecalis from food-producing animals and retail meat.

Increasing numbers of linezolid-resistant Enterococcus carrying optrA are being reported across different niches worldwide. We aimed to characterize the first optrA-carrying Enterococcus faecalis obtained from food-producing animals and retail meat samples in Tunisia. Seven optrA-carrying E. faecalis obtained from chicken faeces (n=3, August 2017) and retail chicken meat (n=4, August 2017) in Tunisia were analysed. Antimicrobial susceptibility was determined by disc diffusion, broth microdilution and Etest against 13 antibiotics, linezolid and tedizolid, respectively (EUCAST/CLSI). optrA stability (∼600 bacterial generations), transfer (filter mating) and location (S1-PFGE/hybridization) were characterized. WGS (Illumina-HiSeq) was done for four representatives that were analysed through in silico and genomic mapping tools. Four MDR clones carrying different virulence genes were identified in chicken faeces (ST476) and retail meat (the same ST476 clone plus ST21 and ST859) samples. MICs of linezolid and tedizolid were stably maintained at 8 and 1-2 mg/L, respectively. optrA was located in the same transferable chromosomal Tn6674-like element in ST476 and ST21 clones, similar to isolates from pigs in Malaysia and humans in China. ST859 carried a non-conjugative plasmid of ∼40 kb with an impB-fexA-optrA segment, similar to plasmids from pigs and humans in China. The same chromosomal and transferable Tn6674-like element was identified in different E. faecalis clones from humans and animals. The finding of retail meat contaminated with the same linezolid-resistant E. faecalis strain obtained from a food-producing animal highlights the potential role of the food chain in the worrisome dissemination of optrA that can be stably maintained without selective pressure over generations.

Exploring the profile of antimicrobial resistance genes harboring by bacteriophage in chicken feces

Bacteriophage may play an important role in antimicrobial resistance genes (ARGs) transmission. However, the contribution of bacteriophage to the spread of ARGs in environment, especially in poultry farm environment, is rarely known. In this study, the prevalence of ARGs in bacteriophage DNA was investigated in chicken feces from 30 different poultry farms in China. Then the abundance of the aac(6′)-Ib-cr, blaCTX-M, ermB, floR, mcr-1, sul1, tetM and intI1 genes was determined by qPCR in bacteriophage and compared with certain representative plasmid DNA samples. The results showed that 12 ARGs (aac(6′)-Ib-cr, aph(3′)-IIIa, blaCTX-M, ermB, ermF, floR, mcr-1, qnrS, sul1, sul2, vanA, tetM genes) and class 1 integron gene intI1 were detected in bacteriophage DNA fraction. The sul1, tetM and aac(6′)-Ib-cr genes were most prevalent with high detection rates of 77%, 61% and 55%, respectively. To our best knowledge, this study firstly reported the presence of the mcr-1 gene in bacteriophage DNA derived from farms environments. We found that the gene copy (GC) numbers of the aac(6′)-Ib-cr, ermB and sul1 genes were as high as 5.47, 5.22 and 5.54 log10 GC/g, respectively. Both the prevalence and abundance of ARGs in broiler fecal wastes were also generally higher than in laying hens. In addition, although the GC numbers of the aac(6′)-Ib-cr, floR and tetM genes in plasmid DNA was higher than that in phage DNA fraction by 4.68, 3.59 and 3.9 orders of magnitude, respectively, the absolute abundances of the blaCTX-M and mcr-1 genes in phage DNA were close to or even higher than that in plasmid DNA at farm SIL2, SIL4 and SIB1. As potential vessels for ARGs, bacteriophage could not be ignored due to their unique extracellular persistence in environments. Overall, this is the first comprehensive survey about bacteriophage carried ARGs from farms in different regions in China.

Detection of virulent thermophilic Campylobacter species in communal chickens

Campylobacter is a foodborne pathogen found in the gut of poultry and other animals, which thereby act as reservoirs for human infection. Campylobacter produces various virulence factors which contribute to the establishment of disease. The aim of this study was to detect virulence genes in Campylobacter isolated from chicken faeces. A total of 408 poultry cloacal samples were analysed through culture, PCR and DNA sequencing. Campylobacter were detected in 24 (5.9 %) of the samples, of which 21 were C. jejuni while 3 were C. coli. The cdtB gene was detected in all Campylobacter isolates while cdtA and cdtC genes were detected in 20 (83.3%) and 18 (75.0%) of the isolates, respectively. The cadF gene was detected in 18 (85.7%) C. jejuni but not in C. coli. Eight genotypes designated G1 to G8 were detected among the isolates. Genotypes G1 and G7 possessed all three cdt genes, a potential for producing functionally active toxins. A large proportion (70.8%) of the isolates possessed both genes for toxin production and adhesion. The high prevalence of cdt and cadF genes in Campylobacter species in this study indicates the pathogenic potential of the isolates. There is a need to implement control measures to curb the spread of these virulent strains.
 Significance:
 
 Virulent thermophilic campylobacters cause severe health complications in humans.
 Although chicken meat is the main source of human infections, there is limited data on the occurrence of Campylobacter in chicken faeces in southern Africa.
 This study adds to literature on the occurrence of virulent strains of Campylobacter species, notably through the deposition of the DNA sequences of our local strains into GenBank®.
 Communal chickens may serve as a source of virulent thermophilic campylobacters to humans, resulting in serious public health implications.
 
 Open data sets: 
 MF872608 - https://www.ncbi.nlm.nih.gov/nuccore/MF872608.1 MF872609 - https://www.ncbi.nlm.nih.gov/nuccore/MF872609.1MF872610 - https://www.ncbi.nlm.nih.gov/nuccore/MF872610.1MF872611 - https://www.ncbi.nlm.nih.gov/nuccore/MF872611.1MF872612 - https://www.ncbi.nlm.nih.gov/nuccore/MF872612.1

Diversity of Essential oils in Sop Cop Nature Reserve in Sơn La province

According the results of the plant species for essential oil in Sop Cop Nature Reserve Area have identified 228 species, 158 genus of 66 families belong Pinophyta and Magnoliophyta. The diversity of essential oils plants fully expressed in 5 life forms with the spectrum of life forms is: SB = 59,65Ph + 13,59 Th + 11,40Cr + 7,89 Ch + 7,44Hm. In addition to the value for essential oils, the species also give other uses such as medicine with 228 species, edible with 64 species, timber with 62 species, making wine yeast with 27 species, spices with 25 species, making bonsai 9 species, making flavor 5 species, 3 species of oil, and lowest with 2 species having poisonous. We have been identified 16 species of plants (accounting for 7.02%) as being endangered in Vietnam Red Book (2007) and 1 species in red list of IUCN (2017). These are species with a small number of individuals that need to have policies for conservation and development
 Keywords
 Essential oil plats, life form, plants, Sop Cop nature reserve.
 References
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Culture Condition for High Biomass Production of Probiotic Yeast Saccharomyces Boulardii SB2 in Shake Flask and 30 Littre Scale

Probiotic properties in vitroof the Saccharomyces boulardii SB2 yeast, was isolated from fruit at Cuc Phuong National Park were evaluated based on biological characteristics and tolerance in simulated intestinal conditions. In the present work, studies were carried out to improve cell growth kinetics to produce cell mass of this biotherapeutic yeast in shake flaskand 30-L bioreactor levels. In case of shake flask, the highest biomass was obtained under the culture conditions as YM medium, at 37oC, pH 6, inoculum concentration of 5 %, with shaking for 32 h; the best sources of carbon and nitrogen were found to be glucose and mixture of peptone, yeast extract, malt extract; cell dry weight and cell density reached 2.43 g L-1 and 109 CFU/ml repetitively. During 30-L bioreactor cultivation, the maximal cell mass in agitation speed of 200 rpm, aeration rate of 1.2 v v-1 min-1; at 32 h of incubation, was 3.58 g L-1 in pH controlled culture and 3.08 g L-1 in uncontrolled pH. This is considered as the first step for cell mass production of this probiotic yeast in industrial scale
 Keywords:
 Saccharomyces boulardii SB2, probiotic, biomass, shake flask, 30-L bioreactor.
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Infectious phage particles packaging antibiotic resistance genes found in meat products and chicken feces

Bacteriophages can package part of their host’s genetic material, including antibiotic resistance genes (ARGs), contributing to a rapid dissemination of resistances among bacteria. Phage particles containing ARGs were evaluated in meat, pork, beef and chicken minced meat, and ham and mortadella, purchased in local retailer. Ten ARGs (blaTEM, blaCTX-M-1, blaCTX-M-9, blaOXA-48, blaVIM, qnrA, qnrS, mecA, armA and sul1) were analyzed by qPCR in the phage DNA fraction. The genes were quantified, before and after propagation experiments in Escherichia coli, to evaluate the ability of ARG-carrying phage particles to infect and propagate in a bacterial host. According to microbiological parameters, all samples were acceptable for consumption. ARGs were detected in most of the samples after particle propagation indicating that at least part of the isolated phage particles were infectious, being sul1the most abundant ARG in all the matrices followed by β-lactamase genes. ARGs were also found in the phage DNA fraction of thirty-seven archive chicken cecal samples, confirming chicken fecal microbiota as an important ARG reservoir and the plausible origin of the particles found in meat. Phages are vehicles for gene transmission in meat that should not be underestimated as a risk factor in the global crisis of antibiotic resistance.

A study of the relationship between human infection with avian influenza a (H5N6) and environmental avian influenza viruses in Fujian, China

BackgroundAvian influenza A (H5N6) virus poses a great threat to the human health since it is capable to cross the species barrier and infect humans. Although human infections are believed to largely originate from poultry contaminations, the transmissibility is unclear and only limited information was available on poultry environment contaminations, especially in Fujian Province.MethodsA total of 4901 environmental samples were collected and tested for Avian Influenza Virus (AIV) from six cities in Fujian Province through the Fujian Influenza Surveillance System from 2013 to 2017. Two patient-related samples were taken from Fujian’s first confirmed H5N6 human case and his backyard chicken feces in 2017. Chi-square test or Fisher’s exact probability test was used to compare the AIV and the viral subtype positive rates among samples from different Surveillance cities, surveillance sites, sample types, and seasons. Phylogenetic tree analysis and molecular analysis were conducted to track the viral transmission route of the human infection and to map out the evolutions of H5N6 in Fujian.ResultsThe overall positive rate of the H5 subtype AIVs was 4.24% (208/4903). There were distinctive differences (p < 0.05) in the positive rates in samples from different cities, sample sites, sample types and seasons. The viruses from the patient and his backyard chicken feces shared high homologies (99.9–100%) in all the eight gene segments. Phylogenetic trees also showed that these two H5N6 viruses were closely related to each other, and were classified into the same genetic clade 2.3.4.4 with another six H5N6 isolates from the environmental samples. The patient’s H5N6 virus carried genes from H6N6, H5N8 and H5N6 viruses originated from different areas. The R294K or N294S substitution was not detected in the neuraminidase (NA). The S31 N substitution in the matrix2 (M2) gene was detected but only in one strain from the environmental samples.ConclusionsThe H5 subtype of AIVs has started circulating in the poultry environments in Fujian Province. The patient’s viral strain originated from the chicken feces in his backyard. Genetic reassortment in H5N6 viruses in Fujian Province was indicated. The H5N6 viruses currently circulating in Fujian Province were still commonly sensitive to Oseltamivir and Zanamivir, but the resistance against Amantadine has emerged.

Morphologic and genomic characterization of a broad host range Salmonella enterica serovar Pullorum lytic phage vB_SPuM_SP116

For effective use of phages as antimicrobial agents for controlling multidrug resistant S. Pullorum, it is important to understand phage biology. A lytic S. Pullorum phage was isolated and characterized from chicken feces, and its whole genome was sequenced and analyzed. A new lytic phage-vB_SPuM_SP116 (in brief SP116)- isolated and characterized using S. Pullorum SPu-116 as its host belongs to Myoviridae A1 group. Phage SP116 had a lytic effect on 27 of 37 (72.9%) different serotypes of clinical Salmonella strains. It showed a high bactericidal activity in killing all pathogens in cultures containing 5 × 105 cfu/mL and achieved more than 6.58 and 5.97 log unit reductions in cultures containing 5 × 106 cfu/mL and 5 × 107 cfu/mL, respectively. The one-step growth curve showed that the burst size was up to 118 pfu/bacterial cell. Complete genome sequence analysis revealed a linear, double-stranded DNA genome of 87,510 bp with an average G + C content of 38.84%, including 128 predicted open reading frames (ORFs) and 22 tRNA genes. SP116 was classified as a Felix O1 virus based upon the general phage characterization and the genomic information. Regarding its high efficacy in preventing especially S. Pullorum infection and its lack of any bacterial virulence, antimicrobial resistance, and lysogenesis genes, it could be a potential alternative candidate for the treatment of S. Pullorum infections.

In Vitro Evaluation of Hen Eggshells Addition in Rice Husk on Litter Properties

Abstract. Ammonium (NH3) is released from uric acid of chicken feces. Good management practices are required to depress ammonium concentration in a litter. Several research findings showed that application of CaCO3 (calcium carbonate) can decrease ammonium concentration in a litter. Eggshell as a by-product of hennery (hen stable) and hatchery is mainly constructed from CaCO3. This research aimed to evaluate the addition of hen eggshell powder at different concentration in litter on water content, pH, microorganism, and NH3 concentration of litter. Rice husk as litter material, hen eggshell powder at a concentration of 0%, 5%, 10%, and 15% and hen manure were thoroughly mixed. After 24 hours, the concentration of water content, pH, microorganisms amount and NH3 in each treatment were measured. This research was completely randomized designed with 3 repetitions. Results showed the progressive addition of hen eggshell powder in litter until 15% increased pH and decreased water content, number of microorganisms and NH3 concentration. The addition of 15% hen eggshell powder in litter revealed best litter properties.Keywords: ammonium, hen eggshell, litter, pH, water content