Sequencing and In Silico Multi-aspect Analysis of S1 Glycoprotein in 793/B Serotype of Infectious Bronchitis Virus Isolated From Iran in 2003 and 2011

Infectious bronchitis (IB) is an acute, highly contagious, and economically important viral disease of chickens. The S1 subunit from Spike (S) protein plays the major role in protective immunity and is involved in the host-virus interactions, as well as infectious bronchitis virus (IBV) serotyping. Aim of the present study was multi-aspect analysis of the molecular and immunological features of 5' part belonging to the S1 glycoprotein sequence of Iranian 793/B IBV strain isolates. This might ideally help in characterization, prevention, and vaccine development. The tissue samples were prepared, followed by virus isolation, reverse transcription polymerase chain reaction and restriction fragment length polymorphism analysis. In addition, sequencing and registration of the sequences in the National Center for Biotechnology Information were performed. Moreover, 12 sequences were retrieved from Fars province, Iran. The next steps included evaluation of conservation/variability along the sequences, phylogenetic analysis, estimation of the average evolutionary divergence over all the sequence pairs, predicting the phosphorylation/N-glycosylation/palmitoylation sites, and the final analysis of antigenicity. The findings of alignment, entropy plot, and pairwise similarity analysis revealed 17 hypervariable regions. The isolates belonging to Tehran were clustered in phylogenetic tree, and the most similar isolates to them were ADW11182 and ADW11183. Location of some of the N-glycosylation/phosphorylation/palmitoylation points indicated that these sites were conserved among the isolates. Furthermore, the frequency of epitopes and their scores reflect the high immunogenicity of S1 protein in 793/B serotype. Analysis of the primary and secondary structures demonstrated that their parameters had variable values and were different regarding the number and location of α-helix, β-strand, and coils. According to our findings, the Iranian isolates of 793/B serotype change their molecular characteristics during time and in different geographical regions. These alterations might account for failure in prevention programs and differences in virulence and pathogenicity

Effect of melatonin supplementation in the long-term preservation of the sheep ovaries at different temperatures and subsequent in vitro embryo production

Investigations in the past decades have shown that oocytes developmental competence following in vitro fertilization is greatly influenced by an interval between isolation of the ovaries immediately after death/slaughter and oocytes recovery from the visible follicles. In order to determine the optimal conditions for long-term preservation of ovaries, an experiment was conducted with adding different doses of melatonin (0 (C), 500 (M1), 600 (M2), 700 (M3) and 800 (M4) μM) as an antioxidant to sheep ovaries preservation medium (PBS) maintained at 4 and 20 °C for 24 h. The effects on in vitro embryo production (IVEP) parameters including maturation, fertilization, cleavage, and blastocyst rates and the total number of blastomere were evaluated after the ovaries preservation. Melatonin reduced the decline in fertilization rate as an indicator of success in vitro maturation (P ≤ 0.05). Furthermore, ovarian storage time had significant negative effect (P ≤ 0.05) on IVEP parameters. Supplementation with melatonin increased the total cell number of blastocysts as an indicator of embryo quality (i.e. mean blastomeric cells in 4Math Eq groups: 86.00 ± 3.00, 98.50 ± 3.5, 111.5 ± 1.5, 125.5 ± 2.00 and 126.50 ± 5.5 for C, M1, M2, M3 and M4. respectively). Overall, the results showed that the use of melatonin antioxidant in the ovaries storage medium had beneficial effects on sheep oocytes development and embryos quality

Effect of oral administration of pioglitazone on follicular dynamics in Holstein dairy cows

This study investigated the effects of oral administration of pioglitazone (PGT), a specific and synthetic ligand of peroxisome proliferator-activated receptors gamma (PPARγ), on follicular dynamics and corpus luteum (CL) functionality in dairy cows. Cows exhibiting strong signs of estrus after 2 injections of PGF2α (given14 d apart) at d 30 postpartum (n = 28) were allotted to four groups (n = 7 cows/treatment) and orally received 6 mg PGT/kg body weight/day according to the following protocol: no PGT (control); PGT for 14 d from 7 d before expected estrus (10 d after 1st injection of PGF2α) to 7 d after observed estrus (PGT14); PGT for 21 d after observed estrus (PGT21); and PGT for 28 d, 7 d before expected estrus to 21 d after observed estrus (PGT28). During the first follicular wave, number of follicles (total and small) increased in PGT14 and PGT28 cows compared to the control group (P < 0.05). During the ovulatory wave, number of total and small follicles increased in PGT28 (P < 0.05) and PGT21 (P < 0.10) compared with PGT14 and control cows. Size of the largest follicle at first wave was greater in PGT28 (P < 0.05), PGT14 (P < 0.05) and PGT21 (P < 0.10) compared to the control cows. Maximal size of the ovulatory follicle was greater in PGT28 (P < 0.05) and PGT21 (P < 0.10) groups compared to the control group. Growth rate of the largest follicle at first wave was higher (P < 0.05) in PGT-treated cows, while growth rate of the ovulatory wave was higher in PGT28 and PGT21 groups, leading to shorter days from luteolysis to ovulation. Pioglitazone administration did not affect CL size, but increased progesterone (P4) concentration. The PGT14 and PGT28 cows had higher maximal plasma P4 concentration and shorter intervals to reach maximal plasma P4 compared to the control group. In conclusion, oral administration of PGT had some positive effects on follicular development and circulating P4 levels which may be conducive to better reproductive performance