Human body is a complex system that is affected by a significant number of microscopic organisms called the microbiomes. The dynamic development of science has led to innovative discoveries in the field of microbiology. This in turn has extracted new field, metagenomics, thanks to which it became possible to perform detailed analysis of individual groups of bacteria and to determine their effects on preserving a good health. One of the biggest scientific projects that would investigate the influence of microbiomes on humans is HMP (Human Microbiome Project). As part of it the research is being conducted leading to characterize human microbiome at the level of nucleotide sequence of the entire genomic DNA. The microflora of the skin, oral cavity, respiratory tract, digestive tract (intestines), genitourinary system has an essential role in the homeostasis. In the last year the carried research proved that it is a vital part of the human organism in preserving a good health. Any changes in its composition may lead to systemic diseases. Pathological changes affect the outcome of the interaction within the microflora that includes species of commensal and pathogenic bacteria, as well as immunology and genetics of the host. Metagenomics research will contribute not only to the recognition of new, so far unidentified by the bacteriological methods microorganisms, but most of all they will serve as a basis to understand the relationships between the human organism and in-dwelling microorganisms. Thanks to the development of the metagenomics or the NGS (Next Generation Sequencing) it will be possible to discover new metabolic pathways and bidirectional links of bacteria with human metabolism. This will help in finding new therapeutic methods in the treatment of many noninfectious diseases so far considered as civilization diseases or genetically conditioned.
Prosthetic joint infections due to Pasteurella multocida are rarely but increasingly reported but no data on production of biofilm are available. We report the case of a woman with a late, haematogenous peri-prosthetic infection of cemented total knee arthroplasty caused by a strain of P. multocida identified by pyrosequencing and unable to produce biofilm. Comparison of clinical and laboratory findings with those reported in other patients evidenced differences mainly in the period of symptoms' onset and in the behaviour of some inflammatory markers.
Trypanosoma evansi infection in the Philippines is frequently reported to affect the country’s livestock, particularly, the buffaloes. To assess the prevalence and intraspecific diversity of T. evansi in the country, blood samples from water buffaloes in different geographical regions were collected during an outbreak. T. evansi was detected in all 79 animals tested using PCR targeting the RoTat 1.2 VSG gene. Sequencing of the rDNA complete internal transcribed spacer (ITS) region including the 5.8S subunit showed high similarity (99–100%) between Philippine isolates and known T. evansi isolates in Genbank. Tree construction based on the same region confirmed the close relationship between Philippine and reported Thai isolates as compared to Egyptian isolates separated by relatively small genetic distances, 47 polymorphisms, despite the clustering in four branches. Overall, the results of this study prove genetic diversity within T. evansi species despite previous reports on limited heterogeneity among isolates worldwide.
The rare tropical Gasteromycetes Lycogalopsis solmsii has been found twice at thirty years interval in the Singapore Botanic Gardens. From the most recent find a culture could be isolated, which allowed DNA extraction and sequencing of about 2000 bp from the nuclear ribosomal DNA. Comparison to a large sample of Basidiomycetes was only possible for a part of the large ribosomal subunit, but clearly indicated affiliation to the gomphoid-phalloid group, without any relationship to Lycoperdales or Agaricales, as stated in the Dictionary of the Fungi.
Next-generation sequencing (NGS) is a novel method widely used in animal science and veterinary research. This technique revolutionized molecular biology and animal genetics research. The unquestionable advantage of NGS is an almost unlimited insight into genetic information. This review discusses the most important applications and achievements in poultry genomics available due to detailed sequence information. Here we present the development of sequencing methods and their further applications in poultry research. The chicken is an important livestock species and a model organism. It is the first non-mammalian amniote whose genome was sequenced by the International Chicken Genome Sequencing Consortium. Therefore, analysis of the chicken genome as a model organism and comparative analysis of genome reference plays an important role in current research. The detailed knowledge of the chicken genome position of genes associated with most important phenotypic traits will contribute to the development of molecular methods for the selection of animals.
Next-generation sequencing (NGS), also known as high-throughput sequencing, is a novel method widely used in animal science and veterinary research. Introduction of NGS was a great breakthrough that has revolutionized the world of molecular biology techniques and animal genetics. An unquestionable advantage of NGS is connected with the almost unlimited insight into genetic information it provides. This review discusses the most important applications and achievements in animal genomics thanks to detailed sequence information. Here we present the history of sequencing and its further development. Applications of the NGS technique in whole genome sequencing, whole exorne sequencing, targeted sequencing of DNA fragments and RNA sequencing in animal research are discussed.
A simplified cDNA cloning protocol was elaborated, easily scalable to very different sizes of plant tissue. The mRNA fraction was extracted from a single wheat kernel with oligo dT magnetic beads. The solid-phase mRNA was transcribed into single-strand cDNA by reverse transcription. Then DNA tags were incorporated into the ds cDNA fragments prior to PCR amplification. The amplified DNA was ligated to a T overhang cloning vector and some of the resulting clones were sequenced. These sequences were identified by BLASTN search in wheat EST databases.
Daring a study on Trypanosoma equiperdum propagation in adult male Wistar rats, four rats died spontaneously. Two of the dead animals were subjected to post mortem examination. From different organs of the rats, chlamydiae were isolated and confirmed by PCR and DNA sequencing as Chlamydophila psittaci. The results show that infection with Chlamydophila psittaci may occur in laboratory rats. Such outbreaks may have influence on the results of experimental studies. Chlamydial infections in laboratory animals also pose risk to humans (zoonosis).
Visfatin is a peptide that is predominantly expressed in visceral adipose tissue and is hypothesized to be related to obesity and insulin resistance. In this study, a novel silent single-nucleotide polymorphism (SNP) was found in exon 7 of the chicken visfatin gene (also known as PBEF1) by single-stranded conformation polymorphism (SSCP) and DNA sequencing. In total, 836 chickens forming an F₂ resource population of Gushi chicken crossed with Anka broiler were genotyped by Xbal forced RFLP, and the associations of this polymorphism with chicken growth, carcass characteristics, and meat quality were analyzed. Significant associations were found between the polymorphism and 4-week body weight (BW4), 6-week body weight (BW6), 4-week body slanting length (BSL4), fat bandwidth (FBW), breast muscle water loss rate (BWLR) and breast muscle fiber density (BFD) (P < 0.05), as well as 4-week breastbone length (BBL4) (P < 0.01). These observations suggested that the polymorphism in exon7 of the visfatin gene had significant effects on the early growth traits of chicken.
In this study we investigated phylogenetics of Miniopterus schreibersii schreibersii and M. s. pallidus from Asia Minor by means of two mitochondrial DNA markers, NADH dehydrogenase subunit 2 (ND2) and cytochrome-b) (Cytb). The average genetic divergence between reciprocally monophyletic M. s. schreibersii and M. s. pallidus was 5.6% on ND2 and 3.5% on Cytb. In all phylogenetic trees, the clade with M. s. schreibersii and M. s. pallidus was placed within Palearctic-Ethiopian Miniopterus taxa. There was a considerable genetic divergence (ca. 8% in Cytb) between M. s. pallidus from Israel and M. s. pallidus from Turkey, Iran, and Nagorno-Karabakh, indicating that they probably are not the same taxon. Time to the most recent common ancestor of M. s. schreibersii and M. s. pallidus was estimated to be between 1.98 to 0.60 Myr BP (ND2 data) and between 1.95 to 0.45 Myr BP (Cytb data).