The results of the metagenomic analysis revealed a high phylogenetic and taxonomic diversity of the microbial community of both luminal and mucosal microflora of the bird intestinal microbiome. Analysis of the taxonomic structure of the compared microbiomes showed both similarities and differences in the taxonomic diversity and relative abundance (%) of bacteria of the compared microbiomes at the level of order, class, genus.At the type level, 24 taxonomic categories were identified in the luminal and mucosal intestinal microbiome of birds, at the class level 35, at the order level 75, at the family level 168, at the genus level 350.The main share of the microbial community of the luminal and mucosal microbiome was made up of microorganisms belonging to the Firmicutes, Proteobacteria, Actinobacteria Cyanobacteria, and Bacteroidetes phyla. Comparison of the biological diversity of the colonic and mucosal microbiomes at the phylum level showed that the relative value of Proteobacteria in relation to Firmicutes in the mucosal microbiome was 3.25 times higher, and the ratio of Firmicutes to Bacteroidetes in the colonic microbiome was 3.98 times higher. than in mucosal.At the genus level, differences in the composition of microbiomes were manifested by a greater relative share (in decreasing order) in the luminal microbiome of representatives of the genus Lactobacillus, Candidatus Arthromitus, Romboutsia, Gallibacterium, Campylobacter, Enterococcus, and representatives of the genus Acinetobacter, Staphylococcus, Bacillus and Bradyrhizobium in the mucosal microbiome. An in-depth study of the intestinal microbiota using cultivation techniques and high-tech methods for identifying microorganisms, taking into account the results of metagenomic analysis, can significantly expand the understanding of the species diversity of microbiomes and the functions of representatives of various taxa in maintaining bird health, as well as to form a census of representatives of normal bird microflora.
Eight laboratories compared counts of Escherichia coli from naturally or artificially contaminated ground beef, other meats and poultry, vegetables, fish and shellfish, cheese, and diverse sources such as swabs, by the Anderson-Baird-Parker direct plate (DP) and a hydrophobic grid-membrane filter (HGMF) method. For five of the eight laboratories overall counts by HGMF were significantly low (51-83%) compared with those by DP. Counts by HGMF tended to be lower for naturally contaminated samples; several possible causes were investigated. In a subsidiary study, analyst variation in counting HGMF ranged from 0.8-7.3%, with little evidence of effects from counting positive versus negative grid cells or from the fullness of growth or staining intensity.
Relevance. Pasteurellosis is a group of zoonotic infectious diseases caused by Pasteurella multocida. According to the antigenic composition, P. multocida is divided into 5 serogroups (A, B, D, F and E). Pathogenic and virulent properties of various serogroups and serotypes of the pathogen in different animal species vary widely and are a marker for determining their role in the development of the disease. Typing of P. multocida strains by capsule groups is an important condition for a comprehensive assessment of the epizootic situation, including for solving the issue of specific disease prevention.Methods. 82 strains of P. multocida from the collection of FGBI «VGNKI», isolated in different years from various animals, were used in the work. Phenotypic typing of pasteurella strains by capsule groups according to Carter was carried out in a test for the detection of hyaluronic acid in a pasteurella capsule and by the type of reaction in a tripaflavin sample. Strains giving a positive reaction with staphylococcus hyaluronidase wereassigned to capsule group A. If the test culture did not belong to group A, but was positive when examined in a tripaflavin sample, it was assigned to capsule group D.Results. Discrepancies between the results of phenotypic typing of pasteurella strains by capsule groups and by PCR were established. The results of typing did not coincide between PCR and Carter typing for capsule group A with respect to 25 strains, for capsule group D with respect to 5 strains and for capsule group B with respect to 5 strains. The number of untyped or doubtful phenotypic properties of pasteurella strains was 29.73%, untyped by PCR 2.46%. Hyaluronidase and acriflavin tests, unlike the PCR-method, do not provide an opportunity for typing pasteurella groups E and F.
Relevance. A significant increase in microbiome-associated diseases, closely related to violations of the bacterial diversity and functions of the normal intestinal microbiota, dictates the need to develop and implement measures for the long-term preservation of individual representatives of the normal microbiota in order to create new strategies for modifying the composition of microbiomes.Methods. The influence of the technology of deep freezing and storage of intestinal isolates of lactic acid bacteria of 2 taxonomic groups isolated from poultry in the conditions of the Low-temperature automated storage of biological samples of the Departmental Collection of useful microorganisms for Agricultural purposes of the Russian Agricultural Academy (VKSM) on the MRS culture medium using 10 and 20% glycerin or 10 and 20% sucrose as cryopreservants was studied. The suspensions of the isolates were frozen at - 150 °C for 18 hours and then placed in an automated cryopreservation at -80 °C. Control of samples for safetyResults. The technology of cryofreezing of lactic acid bacteria on MRS-broth using 10 and 20% glycerin or 10 and 20% sucrose as cryopreservants allows preserving the viability, physiological and biochemical properties of intestinal isolates of lactic acid bacteria when stored for 18 months. All the protective media used (MRS-broth with glycerin 10 and 20%, MRS -broth with sucrose 10 and 20%) showed comparable results in the preservation of viability and acid-forming activity of Lactobacillus fermentum-2, Pediococcus pentosaceus 6p-3, Pediococcus pentosaceus 28p-1 isolates. Then the storage of Pediococcus pentosaceus isolate (28p-1) in a given parameter on a protective medium with 10 and 20% sucrose led to a decrease in the activity of acid formation.
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