Insulin-like growth factor-1 gene (IGF-1) is considered as a major candidate gene for the economic traits of animal production. Polymorphism of 5′ flanking region of IGF-1 gene in Barki sheep (n = 91) and its association with wool traits were studied using the polymerase chain reaction coupled with single-strand conformation polymorphism technique (PCR-SSCP), PCR-restriction fragment length polymorphism (PCR-RFLP), sequence analysis and different measurements of wool traits (clean fleece weight and fiber diameter). PCR-SSCP analysis revealed three different banding patterns corresponding with three genotypes frequencies GG (0.25), GA (0.58), AA (0.17). PCR-RFLP and corresponding sequence analysis revealed nucleotide transversion from Guanine (G) to Cytosine (C) at nucleotide position 85 and transition from (G) to Adenine (A) at position 87. This is the first study that recorded two SNPs within the 5′ flanking region of IGF-1 gene in Egyptian Barki sheep, which were submitted to DNA Data Bank OF Japan (DDBJ) with Accession No. LC151463.1. The genotype GG showed positive significant association (P < 0.001) with clean fleece weight (CFW) trait (Odd Ratio = 2.83). By contrast, genotype AA had negative significant association (P < 0.05) with such trait (Odd Ratio = 0.15). On the other hand, fiber diameter (FD) measurements showed no significant association (P > 0.05) with different IGF-1 genotypes. This study adds evidence of the association between IGF-1 gene polymorphism and CFW of wool in Egyptian Barki sheep. Therefore; it is important to consider IGF-1 gene as a candidate gene marker for wool weight traits and it should be identified before using successful breeding program.

Genetic information especially of the Quantitative Trait Loci (QTL) which affect different performance traits is considered one of the most effective tools in the breeding programs of livestock. Several genes were reported as candidate genes that effect litter size performance and one of these genes is the KISS1 which is considered as a regulator of puberty onset. The polymorphisms of KISS1 gene have some relationships with high prolific and sexual precocity. The objective of this study was the detection of the restriction fragment length polymorphism (RFLP) and single nucleotide polymorphisms (SNPs) of KISS1 gene in six major Egyptian small ruminant breeds. The primers used in this study flanked a 377 bp fragment from intron 1 of KISS1 gene in sheep and goat. These PCR amplified fragments were digested with XmnI endonuclease. According to the presence or absence of the restriction site (GAANN^NNTTC) at position 121^122, we genotyped the 122 tested animals as AT (54.92%) and TT (45.08) with the absence of AA genotype. The overall frequencies of alleles A and T were 27.46 and 72.54%, respectively. The sequence analysis of purified PCR products representing these two detected genotypes declared the presence of a SNP (T→A) at position 125 in the amplified fragment which is responsible for the elimination of the restriction site and consequently the presence of two different alleles T and A. The nucleotide sequences of sheep KISS1 alleles T and A as well as goat KISS1 alleles T and A were submitted to GenBank database and have accession numbers: KP835797, KP835798, KP835799 and KP835800, respectively. It is concluded that small ruminant breeds have high frequency of KISS1 allele T which was associated with greater litter size. We recommend to increase this allele in Egyptian small ruminant breeds and also to select the animals which possess TT genotypes of KISS1 gene and enter them in breeding programs of Egyptian small ruminants to increase their fecundity traits.

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Background No doubt that the corpus luteum (CL) plays a vital role in the regulation of female cyclicity in mammals. The scenarios among microRNAs (miRNAs) and their target genes and steroid hormones {estradiol (E2) and progesterone (P4)} are required for better understanding the molecular regulation of CL during its formation, maturation, and regression. We aimed to (I) study the changes in the relative abundance of miR-205, miR-26a-5p, miR-17-5p, and let-7b-5p and their target genes: LHCGR, CASP3, PCNA, AMH, and PLA2G3, during different stages of corpus luteum in Egyptian buffaloes, and (II) and to address different scenarios between steroid concentrations in the serum and the expression pattern of selected miRNAs and their targets. Methods The paired ovaries and blood samples were collected from apparently healthy 50 buffalo cows at a private abattoir. The ovaries bearing CL were macroscopically divided according to their morphological structure and color into hemorrhagic (CLH), developing (CLD), mature (CLM), regressed (CLR), and albicans (CLA). Small pieces from different stages of CL (CLH, CLD, CLM, CLR, and CLA) were cut and immediately kept at − 80 °C for total RNA isolation and qRT-PCR. The serum was separated for steroid level estimation. Results The LHCGR was expressed during different stages of CL, and the peak of expression was at the mid-luteal stage. The CASP3 revealed a stage-specific response at different stages of CL. The PCNA has an essential role in cellular proliferation in buffaloes CL. Both expression patterns of PLA2G3 and AMH were found over the various developmental and regression stages. It was noticed that miR-205 is conserved to target LHCGR and CASP3 transcripts. Moreover, CASP3 and AMH were targeted via miR-26a-5p. Additionally, the CASP3 and PLA2G3 were targeted via let-7b-5p. The P4 level reached its peak during CLM. There were positive and negative strong correlations between miRNAs (miR-26a-5p and miR-205), target genes (LHCGR and CASP3) during different stages of CL, and steroid hormones in the serum. Conclusions Taken together, the orchestrated pattern among miRNAs, target genes, and steroid hormones is essential for maintaining the proper development and function of CL in buffalo cows.

T HIS study investigated the genetic polymorphism of osteopontin (OPN) gene in Egyptian buffalo bulls in a trial for association with the quality of fresh semen. A total of 228 fresh semen ejaculates were collected from 57 buffalo bulls. Checking the fertility potential, the ejaculates were evaluated for volume, individual motility, live sperm, sperm abnormalities and concentration. The bulls' semen was grouped according to the individual motility into high (>60%, n=47) and low (<60%, n=10) quality. A fragment of 250 bp from intron 3 of OPN gene was amplified by PCR then genotyped by restriction fragment length polymorphism, single strand conformation polymorphism and DNA sequencing. The high-quality semen was significantly increased in individual motility % (p<0.0001), live sperm % (p<0.0001) and sperm concentration (p<0.05) than the low quality. Also, the monomorphic pattern of OPN gene in both groups of bulls was noticed. However, five mutations were discovered including three nucleotide insertions (T 44, A49 and A62) and two nucleotide substitutions (G11>C and A108>G) when comparing the sequence with that of buffalo in the GenBank. In conclusion, OPN gene might have no genetic variation and so it is not associated with semen quality in Egyptian buffalo bulls. Future studies with a larger number of populations on different regions of OPN gene are recommended.

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Growth hormone (GH) gene has been described as a candidate gene for marker-assisted selection in different farm animals. The present study was designed to identify the polymorphism in GH gene and its association with variation of wool traits in Egyptian sheep breeds. Wool and blood samples were collected from 42 animals including two breeds (Barki and Rahmani) and one crossbred (Rahmani x Awase). Measurements of wool traits were analyzed and involved staple strength (Str), staple length (STL), fiber diameter (FD) and clean fleece yield (CFW). DNA was extracted from blood samples and a 365-bp fragment from exon V was amplified by polymerase chain reaction (PCR). Single strand conformation polymorphisim (SSCP) analysis showed two conformational patterns. The pattern I was recorded to be more frequent (83.3, 92.86 and 90%) than pattern II (16.7, 7.14 and 10%) in Barki, Rahmani and crossbred, respectively. The sequence analysis showed one single nucleotide polymorphism (C/T). The pattern I (allele T) has been found to affect CFW and FD than pattern II (allele C). Whereas, C allele was more pronounced for Str and STL. These traits are the most important parameters determining commercial values of wool that are preferred for clothing or carpets industry. The nucleotide sequences of C and T alleles were submitted to GenBank and have the accession numbers: KT250511 and KT250512, respectively. In conclusion, the present results provide evidence that there is a single nucleotide polymorphism within GH gene in Egyptian sheep breeds. This mutation was found to have some effects on wool traits. Therefore our data show interesting prospects in future selection programs for improving wool industry.

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