Journal of Animal Production, Print ISSN: 2008-6776., Online ISSN: 2382-994X

Abstract Objective: Fertility is a fundamental factor for the profitability and sustainability of sheep breeding companies and is also a valuable selection criterion in genetic improvement studies. The Romanov sheep is one of the best-known breeds worldwide in terms of fertility. Excellent results regarding the fertility of Romanov sheep have attracted the attention of many breeders. It seems essential to apply reproductive technologies to Romanov sheep in Iran, which have shown a seasonal decline in reproductive performance. The application of proper management methods during the non-breeding season to increase economic efficiency, especially in imported breeds, is of high importance. This study aimed to evaluate the effect of using short-term 5-day estrus synchronization protocols with two sponge brands containing different doses of eCG hormone on the reproductive performance of Romanov ewes during the non-breeding season. Method: In this study, 90 Romanov ewes aged 2 to 5 years with an average weight of 38.8±1.2 kg were used. The ewes were synchronized for 5 days using either the Esponjavet sponge containing 60 mg medroxyprogesterone acetate (45 ewes) or the Fluorogest sponge containing 40 mg flurogestone acetate (45 ewes). Upon sponge removal, each group was subdivided into three subgroups of 15 ewes each and received 300, 450, or 600 international units of eCG hormone. Results: The Esponjavet sponge significantly reduced mucosal secretions and vaginal adhesions in grades 1 and 2 compared to the Fluorogest sponge (P<0.05). Fertility, lambing, and prolificacy rates were higher in the group treated with Esponjavet sponge combined with 600 IU eCG (P<0.05), while twinning rate, abortion, and barren ewes were not affected. The highest serum progesterone concentration within 48 hours after sponge removal was observed in the Esponjavet treatment combined with 300 IU eCG. In treatments receiving 450 and 600 IU eCG, estrogen levels at 24 and 48 hours post-sponge removal were significantly higher (P<0.05). Cost-benefit analysis showed that the use of the Esponjavet sponge with 600 IU eCG yielded the highest profitability. Conclusion: The results indicated that the reproductive performance of short-term sponge treatments from the two similar brands was comparable. Treatments receiving 600 and 450 IU eCG had similar effects on improving fertility. Economically, the use of the Fluorgest sponge is more cost-effective, and 450 IU eCG combined with this sponge can be used to improve reproductive efficiency and economic profit in breeding flocks. However, if the goal of estrus synchronization protocols is to increase twinning rate, the best option is the Esponjavet sponge with 600 IU of eCG.

Abstract Introduction: Myostatin (MSTN) is a member of transforming growth factor-β (TGF-β), which is a negative regulator for muscle differentiation and growth in various mammals and plays a key role in muscle growth and meat quality. Today, CRISPR technology can be used to accurately change any attribute. The CRISPR/Cas9 technology creates double-strand breaks (DSBs) in the target region of DNA, which can be repaired by homology repair (HDR) in the presence of the corresponding homologous repair template or by non-homologous end joining (NHEJ).
Materials and Methods: Guide RNAs (sgRNA) were designed using CRISPOR online software. Eggs collected from 150 Varami sheep were placed in 50-microliter drops of culture medium in an incubator containing 7% CO₂ and 95% humidity. 22-24 hours after IVM, a mixture of two guide RNAs cloned in a CRISPR vector was injected into each oocyte at a concentration of 30 ng with a microinjection microscope. After microinjection, the parthenogenesis method was used to fertilize the eggs. After the activation of the eggs in the wells of the 96-well plate, the bottom of which was covered with cumulus cells and sage medium for eight days in an incubator with a temperature of 38.5 degrees Celsius and 7% CO₂gas, they were cultivated under conditions of maximum humidity. After eight days, the zygotes that had reached the embryonic stage were analyzed with a fluorescent microscope. The embryos of the test group that emitted green light, as well as one embryo from the control group, were individually placed in nine microliters of DNA Lysis to prepare their genomes. They were incubated in a temperature program of one hour at 65 degrees and ten minutes at 90 degrees. In order to investigate the gene editing of the embryos that emitted green light, the PCR products of five greened embryos along with one embryo of the control group were sequenced by the trench method.
Results and Discussion: Finally, 12 sheep embryos were produced, which were analyzed with a fluorescent microscope, and a total of five embryos emitted green light. The green light indicated that they had received the CRISPR/Cas9 technology. Among the five embryos, two of the embryos with guide RNA 1 showed a single nucleotide deletion upstream of PAM. Additionally, two of the embryos showed a single nucleotide deletion in guide RNA 2, while one of the embryos remained unchanged. Sequence analysis of the knockout embryos revealed that 83% of the cells were cut. After creating two types of single nucleotide deletions in different positions of sheep embryos, the effect of this genomic editing was detected by examining the amino acid sequence of the embryos in the control group and those carrying the mutation. It was observed that the deletion of a single nucleotide caused by guide RNA resulted in a change in the genome framework and termination code, leading to a shorter amino acid sequence in the edited sheep compared to the control group. This research marked the first time that laboratory embryos of Varamin sheep genetically manipulated by CRISPR/Cas9 technology were produced.
Conclusion: The nucleotide sequence of MSTN gene in Varami sheep was different from the sequence recorded in NCBI. Five embryos showed CRISPR technology markers. Both of the designed guide RNAs caused mutations in the nucleotide sequence and termination code in the amino acid sequence.

Abstract In order to investigate the effect of vitamin E and vitamin E nanoliposomes on the expression of the  STAR gene in testis and TSPO gene in the ovary of Japanese quail, an experiment using 864 pieces of quail for 10 weeks in a completely randomized design with six treatments, six replications and 24 Breeding quail (16 females and eight males) was performed in each replication. The treatments were fed diets containing different levels of vitamin E (25, 50 and 100 IU per kg of diet) and vitamin E nanoliposomes (25, 50 and 100 IU per kg of diet). The results of this study showed that the effect of the experimental treatments on the expression of TSPO gene in the ovary and STAR gene in the testis was significant (P<0.05). The addition of 50 IU of vitamin E significantly increased TSPO gene expression in the ovary compared to the control treatment (P<0.05). The results also showed that the level of 25 IU nanoliposome of vitamin E increased the expression of TSPO gene in the ovary, which was not significantly different from the control treatment. The use of vitamin E and vitamin E nanoliposome levels significantly decreased the expression of STAR gene in the testis of Japanese quail (P<0.05). According to the results of this study, the addition of 25 IU vitamin E nanoliposomes as well as 50 and 100 IU levels of vitamin E has a significant effect on the expression of TSPO gene in the ovary, which is one of the genes affecting fertility and reproduction.  

Abstract The aim of present study was to investigate the effect of selection against ascites syndrome on different traits body growth, internal organs, and blood parameters in a broiler line. Therefore, by classifying different families based on the frequency of ascites under intense ascites inducing condition (AIC), 10 susceptible families (SUS) and 10 resistant families (RES) to ascites were selected and another set of offspring from the same families were bred under normal commercial condition (NCC). This process of selection and cross validation was carried out twice in the two distinct generations. The genetic relationship of different traits, the difference in the frequency of ascites between AIC and NCC, and the difference between different traits in SUS and RES were performed by correlation statistics, Chi-square test, and GLM procedure of SAS software (version 9.1), respectively. The correlation between ascites susceptibility in different families indicated that families that showed more ascites under AIC also had more ascites under NCC. Comparative study of different traits in susceptible and resistant groups showed that in SUS, body weight, growth rate and respiratory capacity were significantly lower and blood partial pressure of carbon dioxide was significantly higher than RES (P<0.05). In the present study it was shown that selection against this syndrome not only does not reduce growth rate in the investigated broiler line, but also improves growth traits and resistance to this syndrome in the mentioned line.

Abstract The objective of this study was to identify known intergenic lncRNAs related to biological pathways of acidosis in Holstein calves using ruminal
tissue. Two groups of healthy calves (N=3) and affected by acidosis (N=3) were compared. Paired-end sequencing method was performed using the
Hiseq2500 illumine platform. Hisat2 software was used to align reads to the bovine reference genome and StringTie software package was used to
assemble read files into transcripts. Using next generation sequencing, 1636 genes belonging to known intergenic lncRNAs were identified, of which
56 genes showed significant differential expression (P≤0.05). Neighbor genes of known intergenic lncRNAs were determined on bovine genome.
Analysis of biological pathways and molecular function showed that five biological pathways were significantly (P≤0.05) enriched. These pathways
were Apelin signaling pathway, Gap junction, Glucagon signaling pathway, Renin secretion, and AGE-RAGE signaling pathway. Moreover, two
molecular functions including gap junction channel activity, and phosphatidyl inositol phospholipase C activity were significantly (P≤0.05) enriched.
Some lncRNAs have different expression in healthy and acidosis samples, and the decreased pH acts as a stimulus to activate some biological
signaling pathways. In conclusion, it was indicated that lncRNAs with differential expression between the control group and the group affected by
acidosis are associated with pathways related to rumen energy metabolism and signaling. Identified differentially expressed lncRNAs could be used as
prognostic in acidosis and biomarkers or promising candidates in animal breeding.

Abstract This study was conducted to investigate the Single nucleotide polymorphism of STAT1 gene by PCR-RFLP technique and association of genotypes with milk production traits in Brown Swiss dairy cattle. The blood samples of ninety-eight cows were collected randomly from tail vein. The DNA was extracted with Cinagene kit (PR881612C). Fragment of 314 bp in 3'UTR of STAT1 gene was amplified by PCR and digested by Pag1 restriction enzyme. Genotype frequencies were calculated by PopGene software (version 1.31). The SAS 9.1 software was used for statistical analysis and the mean of traits was compared by Duncan test. The genotype
frequencies of TT, TC and CC were 0.23, 0.25 and 0.52, respectively. The T and C allele frequency were 0.355 and 0.645, respectively. The expected heterozygosity was estimated 0.458. Result of Chi-square showed that this locus was not in the Hardy Weinberg equilibrium (P<0.05). The means of traits for milk, fat percent and fat yield, and protein yield were not significant but means of protein percent was significant (P<0.05). The mean of milk, fat and protein yield traits for CC genotype were more than two other genotypes. The results of present study showed that T allele is known as minor frequency allele and C allele have positive effect on milk, fat and protein yield traits.

Abstract Genetic parameters direct and maternal in Fars native flows using the Bayesian statistical approach, was estimation with consider body weight at a day of age (BW₁), body weight at eight weeks of age (BW8), body weight at 12 weeks of age (BW₁₂), age at sexual maturity (ASM), egg number (EGP) and mean egg weight during 28^th, 30^th and 32^nd week ages (AV283032) Fars native flows. Genetic parameters were estimated by six different animal models and MTGSAM software. The best model was determined using the Akaike information criterion for each trait. The direct heritability (h²) estimates basis best model BW₁, BW₈, BW₁₂, ASM, EGP and AV283032 0.36, 0.33, 0.30, 0.48, 0.26 and 0.25, respectively. Maternal effects were significant for ASM, EGP and AV283032 traits studied and estimates of maternal environmental variance as a proportion of phenotypic variance (c2) varied from 0.01 (EGP and AV283032) to 0.39 (BW₁). Results present research showed that Bayesian approach by considering all factors more complicated models was selected as a model.

Abstract This study was conducted to detect polymorphism of gene BMP15, a member of Transforming Growth
Factor β (TGFβ) family which has a crucial role in controlling the ovarian follicles development,
ovulation rate and fertility. Samples were randomly selected from 91 Najdi goats in 3 geographical
locations, northwest, southeast and center of Khuzestan province. After DNA extraction, amplification of
235 bp fragment of exon 2 of BMP15 gene was performed using specific primers. Sequence detection
was executed after amplification of gene fragments. The association of BMP15 gene and litter size was
done by SAS software. Results from sequencing were analyzed by Vector NTI software. The results
identified three mutations in bases 529 (T to G), 530 (C to G) and 576 (T to C). The largest litter size
belongs to AA pattern. Point mutations in this gene will alter the Ovulation rate of the goat. Therefore,
improving twining trait in Najdi population can be expected by marker assisted selection.