Implication of complex vertebral malformation and deficiency of uridine monophosphate synthase on molecular-based testing in the Iranian Holstein bulls population

The aim of this study was to identify the deficiency of uridine monophosphate synthase (DUMPS) and the complex vertebral malformation (CVM) in Iranian Holstein bulls. A total of 144 blood samples were prepared of Holstein bulls in Abbas Abad Animal Breeding Center and Ferdowsi University of Mashhad's Dairy Farm in Khorasan state of Iran. Genomic PCR-RFLP protocol was performed to amplify the polymorphic region of the bovine uridine monophosphate synthase UMPS gene. Also, genomic PCR-SSCP method was performed for CVM to amplify the polymorphic region of the bovine solutecarrier family 35 member 3 (SLC35A3) genes. The results of this study demonstrated that there was no carrier of DUMPS and CVM in Iranian bulls in these centers

Bioinformatics Analysis of Upstream Region and Protein Structure of Fungal Phytase Gene

Phytase increases the bioavailability of phytate phosphorus in seed-based animal feeds and reduces the phosphorus pollution of animal waste. Since most animal feeds for pellets are heated up to 65-80 °C, the production of a thermostable structure for phytase can be useful. In this study, we sought to perform bioinformatics analysis of the upstream region and protein structure of fungal phytase to improve its expression and thermostability properties. We used bioinformatics methods such as similarity search, multiple alignment, statistical analysis of physicochemical properties of amino acids, pattern recognition, and protein modeling to find out the effective factors in heat resistance of phytase. Change in Gibbs free energy (ΔG) of the best pattern promoter resulting from the interaction between RNA polymerase and the promoter sequences of modified genes of phytase was equal to -9 kcalmol-1, which is lower compared to other interactions. The evaluation of the three-dimensional structure of new phytases showed that amino acid substitutions aimed at improving thermostability did not change the form and structure of the protein. The results of Prochek, Whatcheck, and ERRAT for structural analysis and verification were 84, 72, and 70, respectively, that were satisfactory

Breadth of the CD4+ T cell response to Anaplasma marginale VirB9-1, VirB9-2 and VirB10 and MHC class II DR and DQ restriction elements

MHC class II molecules influence antigen-specific CD4(+) T-lymphocyte responses primed by immunization and infection. CD4(+) T-cell responses are important for controlling infection by many bacterial pathogens including Anaplasma marginale, and are observed in cattle immunized with the protective A. marginale outer membrane (OM) vaccine. Immunogenic proteins that comprise the protective OM vaccine include type IV secretion system (T4SS) proteins VirB9-1, VirB9-2, and VirB10, candidates for inclusion in a multi-epitope vaccine. Our goal was to determine the breadth of the VirB9-1, VirB9-2, and VirB10 T-cell response and MHC class II restriction elements in six cattle with different MHC class II haplotypes, defined by DRB3, DQA, and DQB allele combinations for each animal. Overlapping peptides spanning each T4SS protein were tested in T-cell proliferation assays with autologous antigen presenting cells (APC) and artificial APC expressing combinations of bovine DR and DQ molecules. Twenty immunostimulatory peptides were identified; three representing two or more epitopes in VirB9-1, ten representing eight or more epitopes in VirB9-2, and seven representing seven or more epitopes in VirB10. Of eight DRA/DRB3 molecules, four presented 15 peptides, which was biased as DRA/DRB3*1201 presented ten and DRA/DRB3*1101 presented four peptides. Four DQA/DQB molecules composed of two intrahaplotype and two interhaplotype pairs presented seven peptides, of which five were uniquely presented by DQ molecules. In addition,three functional mixed isotype (DQA/DRB3) restriction elements were identified. The immunogenicity and broad MHC class II presentation of multiple VirB9-1, VirB9-2, and VirB10 peptide epitopes justify their testing as a multi-epitope vaccine against A. marginale

Comparison of bovine lymphocyte antigen DRB3.2 allele frequencies between two subpopulations of Iranian holstein cattle

The bovine lymphocyte antigen (BoLA-DRB3) gene encodes cell surface glycoproteins that initiate immune responses by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the mostpolymorphic bovine MHC class II gene which encodes the peptide-binding groove. Since different alleles favor the binding of different peptides, DRB3 has been extensively evaluated as a candidatemarker for associations with various bovine diseases and immunological traits. Therefore, in this study, the genetic diversity of the bovine class II DRB3 locus in the two Iranian subpopulations of Holstein cattle (Moghan farm n = 250 and Razavi farm n = 175) was investigated by polymerase chain reactionrestriction fragment length polymorphism method (PCR-RFLP). Bovine DNA was isolated from whole blood. A hemi-nested PCR followed by digestion with restriction endonucleases RsaI, HaeIII and BstYI was conducted on the DNA. The results indicated that exon 2 of the BoLA-DRB3 gene is highlypolymorphic in the two populations, and the frequency of BoLA-DRB3 depends on breed. On the other hand the presence of BoLA-DRB3*8, *24, and *16 alleles with high frequency in BoLA-DRB3.2 system,can be used as appellative index for nominate Holstein

The Effect of Cinnamon Extract on Spermatogenesis and

A semi-field study was carried out to evaluate the effect of two medicinal herbs, individually and in combination, on growth performance, carcass traits, nutrient digestibility and immune response of broiler chickens. A total of 384 one-day-old straight-run Arbor Acres broiler chickens were allocated into 24 floor pens prepared in a commercial broiler house. Pen-groups were fed one of the following five diets for 42 days: a basal corn-soybean meal diet as control (5 pens), the same basal diet plus 200 ppm virginiamycin (V; 4 pens), the same basal diet supplemented with 0.4% peppermint (Mentha piperita) leaves (P; 5 pens), 0.4% tarragon (Artemisia dracunculus) leaves (T; 5 pens) or with 0.2% tarragon leaves + 0.2% peppermint leaves (P+T; 5 pens). The results showed that performance traits, including average body weight, body weight gain, feed intake and feed conversion ratio were not affected by dietary treatments (P>0.05). No significant differences were detected between the control and experimental groups in apparent digestibility of nutrients and antibody titer against newcastle disease virus (NDV). Slaughter traits of herb or antibiotic supplemented groups did not differ significantly from those of the non supplemented control group. In conclusion, the additives tested had no impact on broiler growth and health status

Genetic Diversity in Four Microsatellite Loci BMS1915, BMS1350, LGB and ILSTS45 in Baluchi Sheep

The poly morphism of 4 micro satellite loci (BMS1350, LGB, ILSTS45, BMS1915) associated with functional traits were evaluated for detecting the diversity level in Baluchi breed of sheep. One hundred eighty five blood samples were collected from sheep herd in Abbas-Abad breeding station. DNA extraction was performed using guanidinethiocyanate silica gel method. Gene fragments of micro satellites were amplified by polymerase chain reaction based on the recommended standard method. The PCR products were visible using poly acry lamide gel electrophoresis and stain methods with silver. Alleles were typed using the Photocapt software version 3. All loci were in the Hardy-Weinberg equilibrium (

The Effect of Cinnamon Extract on Spermatogenesis and

Genotypes of Iranian Zel sheep for Calpastatin (CAST) locus were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) methods and for Calpain (CAPN) locus by PCR-SSCP. Blood samples were collected from 200 purebred Zel sheep of Zel Breeding Station located in Golestan province in northeast of Iran. Extraction of genomic DNA was based on modified salting out method. The digestion of PCR products of CAST gene by MspI and NcoI restriction enzymes revealed two alleles M and N, with frequencies 85.5 and 14.5%, respectively. Frequencies were 75, 21 and 4% for MM, MN and NN genotypes, respectively. Alternatively, using PCR-SSCP method, four genotypes including AA, AB, BB and AC with frequencies of 71, 21, 4 and 4%, respectively, were observed in this population. Analyzing CAPN gene by the PCR-SSCP method, revealed two different conformational patterns (AA and AB) with frequencies of 69 and 31% for AA and AB, respectively. Average heterozygosity for both loci was low (0.28 and 0.25% for CAST using PCR-SSCP and PCR-RFLP, and 0.26% for CAPN). Yearling weights (YW) were analyzed by a statistical model comprising PCR-SSCP and as a result CAPN genotypes had significant effect (P<0.01) on YW. A Chi-square test confirmed Hardy-Weinberg (H-W) equilibrium for the CAST locus using PCR-SSCP method but not for PRC-RFLP method and CAPN locus. Totally, the investigated herd had little genetic diversity and different factors disturb H-W equilibrium and PCR-RFLP and PCR-SSCP might be used successfully in these studies