A novel B- and helper T-cell epitopes-based prophylactic vaccine against Echinococcus granulosus

Introduction: In this study, we targeted the worm stage of Echinococcus granulosus to design a novel multi-epitope B- and helper T-cell based vaccine construct for immunization of dogs against this multi-host parasite. Methods: The vaccine was designed based on the local Eg14-3-3 antigen (Ag). DNA samples were extracted from the protoscoleces of the infected sheep’s liver, and then subjected to the polymerase chain reaction (PCR) with 14-3-3 specific forward and reverse primers. For the vaccine designing, several in silico steps were undertaken. Three-dimensional (3D) structure of the local Eg14-3-3 Ag was modeled by EasyModeller software. The protein modeling accuracy was then analyzed via various validation assays. Potential transmembrane helix, signal peptide, post-translational modifications and allergenicity of Eg14-3-3 were evaluated as the preliminary measures of B-cell epitopes (BEs) prediction. Having used many web-servers, a well-designed process was carried out for improved prediction of BEs. High ranked linear and conformational BEs were utilized for engineering the final vaccine construct. Possible T-helper epitopes (TEs) were identified by the molecular docking between 13-mer fragments of the Eg14-3-3 Ag and two high frequent dog class II MHC alleles (i.e., DLA-DRB1*01101 and DRB1*01501). The epitopes coverage was evaluated by Shannon’s variability plot. Results: The final designed construct was analyzed based on different physicochemical properties, which was then codon optimized for high-level expression in Escherichia coli k12. This minigene construct is the first dog-specific epitopic vaccine construct that is established based on TEs with high-binding affinity to canine MHC alleles. Conclusion: This in silico study is the first part of a multi-antigenic vaccine designing work that represents as a novel dog-specific vaccine against E. granulosus. Here, we present key data on the step-by-step methodologies used for designing this de novo vaccine, which is under comprehensive in vivo investigations

Effect of three methods of estrus synchronization: Osynch, Co-Synch and Prostaglandin on the fertility of subfertile dairy cows

This study was performed to compare the effect of using 3 different estrus synchronization methods (Ovsynch, Co-Synch and Prostaglandin) on the conception rate of subfertile dairy cows. Fifty one subfertile Holstein dairy cows with parity ranging from 1 to 7 were enrolled in this study. Cows were allocated to three groups Ovsynch (n=10), Co-Synch (n=27) and Prostaglandin (n=14). Cows in the Ovsynch group underwent a standard Ovsynch protocol (GnRH analogue on Day 0, PGF2α analogue on Day 7 and GnRH analogue on Day 9), then they were artificially inseminated approximately 16 hours after the second GnRH treatment. The same procedure was done for cows in Co-Synch group, but artificial insemination took place immediately after the last injection. Cows in prostaglandin group underwent a PG protocol (PGF2α, on Day 0 and 12), and artificial insemination was done about 72 hours after the second PGF2α injection. Pregnancy diagnosis was performed 60 days after insemination via rectal examination. By using logistic procedure of SAS 9.1 software, the effects of treatment, season, parity, animal, age and open days were analyzed. The analysis of variables related to animal and the measured effects on pregnancy proved that none of the evaluated parameters were significant. Hence, we cannot declare which of the estrus synchronization methods is suitable to increase pregnancy rate. Further studies with more replicates are required to choose the best method

Comparative effect of different concentrations of hydro-ethanolic extract of chamomile on freeze-thawn semen quality of rams

Oxidative stress during freeze-thawing process causes reduction in motility, viability, membrane functions and finally sperm fertility. The aim of this study was to determine the effect of Chamomile extract as natural antioxidant on quality of cryopreserved ram sperm. In this study semen was collected from five mature rams twice a week using an artificial vagina and the ejaculates were pooled equally in order to eliminate the individual effects. Different levels of extract of Chamomile (0, 50, 66.66, 100 and 200 ml/dl of diluent solution) were added to diluent based tris-egg yolk. After cooling, filling and sealing of the samples, they were frozen with nitrogen vapor and immersed in liquid nitrogen and were stored until evaluation time. After thawing, results showed that addition of 66.66 ml/dl of extract increased total motility, progressive motility, curvilinear velocity, straight–line velocity, linearity, viability and plasma membrane integrity of sperm compared to the control and other treatment groups (