Application of culturomics to characterize diverse anaerobic bacteria from the gastrointestinal tract of broiler chickens in relation to environmental reservoirs

2020 ◽  
Vol 66 (4) ◽  
pp. 288-302
Author(s):  
Paul E. Moote ◽  
Sarah J.M. Zaytsoff ◽  
Rodrigo Ortega Polo ◽  
D. Wade Abbott ◽  
Richard R.E. Uwiera ◽  
...  

Characterization of the microbiota of chickens is of current interest. The goals of the current study were to apply anaerobic isolation methods to comprehensively isolate and identify bacteria from the gastrointestinal tract of chickens and their environment. Bacterial communities within the drinking water were dominated by Escherichia, whereas communities in litter were more representative of the cecum. The crop and small intestine (jejunum and ileum) were dominated by Lactobacillus and Enterococcus spp., and the cecum was dominated by Proteus spp. The collection of bacteria isolated was dominated by Enterococcus spp., Escherichia/Shigella spp., Lactobacillus spp., and Proteus spp.; however, many rare taxa were observed. These included members of the Clostridiales and Clostridium spp., which were commonly isolated from the ileum and cecum. Bacteria isolated by enrichment and direct plating differed. The selective de Man–Rogosa–Sharpe agar was commonly associated with the isolation of Lactobacillus spp. and yielded the lowest diversity of all methods utilized. Increased diversity and frequency of Clostridium spp. was observed in enrichments of blood and mucus or by plating on Columbia agar supplemented with 10% blood and gentamicin. The bacteria isolated from this study provide source material for genomic and functional studies in chicken hosts.

2018 ◽  
Vol 20 (83) ◽  
pp. 376-380
Author(s):  
R.A. Pelenio ◽  
V.V. Stybel ◽  
V.O. Ushkalov

The changes in lipolytic, lysozyme and hemolytic activity of the intestinal microflora of piglets invaded with the association of ascaris, imyrium and balantidium have been studied. It has been established that among microorganisms isolated from the distal intestine of piglets, lipase is synthesized by Bacteroides spp., Prevotella spp., Clostridium sp. and Peptostreptococcus spp., E. coli lact. «-» hem +, Enterobacter spp., Streptococcus spp., Staphylococcus spp. and Candida spp. In pigs affected by ascaris, emerya, and balantidium, the number of strains producing lipase was higher compared to healthy ones, namely: Bacteroides spp. – by 20.2%, Prevotella spp. – by 3.7%, Clostridium spp. – 29.2% and Peptostreptococcus spp. – by 14.9%, Staphylococcus spp. – by 67.7%, Streptococcus spp. – by 19.1%, E. coli lact. «-» heme «+» – by 17.3%, Candida spp. – 16.5% and Enterobacter spp. – by 4.8%. Lysozyme activity was detected in Bifidobacterium spp., Lactobacillus spp. and Staphylococcus spp. Significant differences in the penetrance of lysozyme activity of microorganisms in healthy and affected by ascarids, imedium, and balantidias of piglets were observed in Lactobacillus spp. and Staphylococcus spp. (P < 0.001). At the same time, in piglets, compared with healthy animals, the number of Lactobacillus spp. Strains that showed lysozyme activity decreased, while Bifidobacterium spp. and Staphylococcus spp. – grew. Bifidobacterium spp., Eubacterium spp., Lactobacillus spp., E. coli lact. «+» Hem «-», Enterobacter spp. and Citrobacter spp. there is no ability to produce hemolysins. Among lactosonegative gastric sticks, all strains isolated from both healthy and sick pigs showed hemolytic activity. The associative influence of ascaris, imerium and balantidii, showed an increase in the penetrance of hemolytic activity in Bacteroides spp. – by 45.8%, Prevotella spp. – by 26.8%, Clostridium spp. – 42.1%, Propionibacterium spp. – 41.5%, Peptostreptococcus spp. – 29.1% and Fusobacterium spp. – by 16.4%, Klebsiella spp. – by 33.4%, Enterococcus spp. – by 69.6%, Streptococcus spp. – by 66.0%, Staphylococcus spp. – by 64.5% and Candida spp. – by 11.5%.


2010 ◽  
Vol 76 (24) ◽  
pp. 8033-8043 ◽  
Author(s):  
Moussa S. Diarra ◽  
Heidi Rempel ◽  
Julie Champagne ◽  
Luke Masson ◽  
Jane Pritchard ◽  
...  

ABSTRACT Enterococci are now frequent causative agents of nosocomial infections. In this study, we analyzed the frequency and distribution of antibiotic resistance and virulence genotypes of Enterococcus isolates from broiler chickens. Fecal and cecal samples from nine commercial poultry farms were collected to quantify total enterococci. Sixty-nine presumptive enterococci were isolated and identified by API 20 Strep, and their susceptibilities to antibiotics were determined. Genotypes were assessed through the use of a novel DNA microarray carrying 70 taxonomic, 17 virulence, and 174 antibiotic resistance gene probes. Total enterococcal counts were different from farm to farm and between sample sources (P < 0.01). Fifty-one (74%) of the isolates were identified as E. faecium, whereas nine (13%), seven (10%), and two (3%) isolates were identified as E. hirae, E. faecalis, and E. gallinarum, respectively. Multiple-antibiotic resistance was evident in E. faecium and E. faecalis isolates. The most common multiple-antibiotic resistance phenotype was Bac Ery Tyl Lin Str Gen Tet Cip. Genes conferring resistance to aminoglycoside (aac, aacA-aphD, aadB, aphA, sat4), macrolide (ermA, ermB, ermAM, msrC), tetracycline (tetL, tetM, tetO), streptogramin (satG_vatE8), bacitracin (bcrR), and lincosamide (linB) antibiotics were detected in corresponding phenotypes. A range of 9 to 12 different virulence genes was found in E. faecalis, including ace, agg, agrBEfs (agrB gene of E. faecalis), cad1, the cAM373 and cCF10 genes, cob, cpd1, cylAB, efaAEfs , and gelE. All seven E. faecalis isolates were found to carry the gelE gene and to hydrolize gelatin and bile salts. Results from this study showed the presence of enterococci of public and environmental health concerns in broiler chicken farms and demonstrated the utility of a microarray to quickly and reliably analyze resistance and virulence genotypes of Enterococcus spp.


2016 ◽  
Vol 3 (1) ◽  
pp. 43-48 ◽  
Author(s):  
V. Patyka ◽  
L. Butsenko ◽  
L. Pasichnyk

Aim. To validate the suitability of commercial API 20E test-system (bioMerieux) for the identifi cation and characterization of facultative gram-negative phytopathogenic bacterial isolates. Methods. Conventional mi- crobiological methods, API 20E test-system (bioMerieux) according to the manufacturer’s instructions. Re- sults. The identifi cation results for Erwinia amylovora, Pectobacterium carotovorum and Pantoea agglome- rans isolates were derived from the conventional and API 20E test systems, which, were in line with the literature data for these species. The API 20E test-system showed high suitability for P. agglomerans isolates identifi cation. Although not all the species of facultatively anaerobic phytopathogenic bacteria may be identi- fi ed using API 20E test-system, its application will surely allow obtaining reliable data about their physiologi- cal and biochemical properties, valuable for identifi cation of bacteria, in the course of 24 h. Conclusions. The results of tests, obtained for investigated species while using API 20E test-system, and those of conventional microbiological methods coincided. The application of API 20E test-system (bioMerieux) ensures fast obtain- ing of important data, which may be used to identify phytopathogenic bacteria of Erwinia, Pectobacterium, Pantoea genera.


2020 ◽  
pp. 16-18
Author(s):  
V. M. Lykhman ◽  
O. M. Shevchenko ◽  
Ye. O. Bilodid ◽  
Igor Vladimirovich Volchenko ◽  
I. A. Kulyk ◽  
...  

Among urgent surgical diseases of abdominal cavity, an acute intestinal obstruction is the most difficult to be diagnosed and treated. Leading factor, determining the development of pathophysiological processes is considered to be the progressive manifestations of enteric insufficiency syndrome, resulting in intestinal barrier impairment, negative changes in ecology of intestinal flora, increased endotoxins. To identify the small intestine microflora in acute intestinal obstruction and determine the role of dysbiotic disorders in clinical manifestations of main pathological process, a study was conducted in 60 patients with mechanical intestinal obstruction. The small intestine has a relatively rare microflora, consisting mainly of gram−positive facultative aerobic microorganisms, streptococci, lactobacilli. The distal ileum in nearly 30−55 % of healthy people contains scanty microflora, and yet the flora of this area differs from the microbial population of the higher gastrointestinal tract due to higher concentration of gram−negative bacteria. Optional−anaerobic coliform bacilli, anaerobic bifidobacteria and fusobacteria, bacteroids, the number of which starts exceeding the one of gram−positive species, are presented in significant quantities. Distal to the ileocecal valve there are significant changes in the microflora quantitative and species composition. Obligatory anaerobic bacteria become the predominant part of microflora, exceeding the number of aerobic and facultative anaerobic bacteria. The bacterial flora in different parts of gastrointestinal tract has its own specifics and is quite constant, as a result of the interaction of many factors, regulating the bacterial population in small intestine. The most important among them are: acidity of gastric juice, normal peristaltic activity of the intestine, bacterial interactions and immune mechanisms. Disorders of the intestine motor and evacuation function with its obstruction lead to slow passage of the chyme and contamination of the upper gastrointestinal tract with new types of microbes. There is a syndrome of small intestine excessive colonization, which means an increased concentration of bacterial populations in it, similar in species composition to the colon microflora. Pathological intra−intestinal contents become a source of endogenous infection and re−infection of the patient, leads to internal digestive disorders, which is manifested by syndrome of malabsorption of proteins, carbohydrates and vitamins. Key words: acute intestinal obstruction, small intestinal microflora, conditionally pathogenic microorganisms, intestinal biocenosis.


Sign in / Sign up

Export Citation Format

Share Document