Allele mining in prolactin receptor gene and its association with some economic traits in Egyptian goat breeds.

The objectives of the current study were to identify polymorphism in the prolactin receptor (PRLR) gene among three Egyptian goat breeds (Zaraibi, Damascus, and Barki) and to investigate the association between PRLR genotype, parity, season of kidding, and litter size factors with milk yield and reproductive traits of Zaraibi goats. One hundred and ninety blood samples were collected for DNA extraction, with 110 from Zaraibi, 40 from Barki, and 40 from Damascus breeds. Three genotypes, CC, CT and TT, for the prolactin receptor gene were identified in the 190 DNA samples using restriction fragment length polymorphism and were confirmed by direct sequencing technique. Milk yield during suckling and lactation periods in addition to age at first conception, gestation length, and litter size were determined in 110 Zaraibi goats. The Zaraibi goats recorded the highest heterozygosity (0.495) and the effective number of alleles (1.972). The g.62130C > T SNP showed a significant association (p < 0.01) with suckling, lactation, and total milk yield of Zaraibi goats with the highest values recorded at the third parity. Age at the first conception and gestation length traits were significantly influenced by the kidding season (p < 0.05) with younger age in autumn and shorter length in spring seasons. Milk yield during the suckling period was significantly (p < 0.01) higher in the case of triplets' litter size. The current study showed that litter size and parity played an important role in the amount of Zaraibi goats' milk yield. The g.62130C > T SNP of the PRLR gene may be a useful marker for assisted selection programs to improve goat milk yield during suckling and lactation periods with the heterozygous genotype CT recording the highest values.

Polymorphism of lactoferrin gene in Egyptian goats and its association with milk composition traits in Zaraibi breed.

The objectives of this study were to identify polymorphisms in the lactoferrin gene among three Egyptian goat breeds (Barki, Zaraibi, and Damascus) and to investigate the effect of LF genotype, parity, and lactation stage on milk composition traits of Zaraibi goats. One hundred and thirty-two blood samples were collected for DNA extraction, with 53 from Zaraibi, 40 from Damascus, and 39 from Barki breeds. Fat, protein, total solids, solids-not-fat, and lactose percentages were determined in Zaraibi goat milk using an automatic milk analyzer. Two genotypes, GG and GA, in the lactoferrin gene were identified using single-strand conformation polymorphism and were confirmed by direct sequencing technique. The Zaraibi breed recorded the highest heterozygosity (0.272) and effective number of alleles (1.369), while the Damascus breed recorded the lowest values. The G/A SNP showed a significant association with protein, solids-not-fat, and total solid content of Zaraibi goat milk. Protein, solids-not-fat, and total solid content in our study were significantly higher at early and late parities. Lactose percentage decreased significantly from early to late parity. Fat, protein, solids-not-fat, and total solid content were significantly higher at early and late stages of lactation, and our results encourage the utilization of Zaraibi goat milk in cheese and butter processing at these stages. Moreover, the G/A SNP of the LF gene may be a useful marker for assisted selection programs to improve goat milk composition.

Biochemical and genotyping analyses of camels (Camelus dromedaries) trypanosomiasis in North Africa.

Camels are considered an important food source in North Africa. Trypanosomiasis in camels is a life-threatening disease that causes severe economic losses in milk and meat production. Therefore, the objective of this study was to determine the trypanosome genotypes in the North African region. Trypanosome infection rates were determined by microscopic examination of blood smears and polymerase chain reaction (PCR). In addition, total antioxidant capacity (TAC), lipid peroxides (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) were determined in erythrocyte lysate. Furthermore, 18S amplicon sequencing was used to barcode and characterizes the genetic diversity of trypanosome genotypes in camel blood. In addition to Trypanosoma, Babesia and Thelieria were also detected in the blood samples. PCR showed that the trypanosome infection rate was higher in Algerian samples (25.7%) than in Egyptian samples (7.2%). Parameters such as MDA, GSH, SOD and CAT had significantly increased in camels infected with trypanosomes compared to uninfected control animals, while TAC level was not significantly changed. The results of relative amplicon abundance showed that the range of trypanosome infection was higher in Egypt than in Algeria. Moreover, phylogenetic analysis showed that the Trypanosoma sequences of Egyptian and Algerian camels are related to Trypanosoma evansi. Unexpectedly, diversity within T. evansi was higher in Egyptian camels than in Algerian camels. We present here the first molecular report providing a picture of trypanosomiasis in camels, covering wide geographical areas in Egypt and Algeria.

Genetic similarity and diversity among three camel populations reared in Egypt.

BACKGROUND: Molecular genetics has been extremely useful in determining the relation between animal populations and documenting the degrees of genetic variation found within them. The present study was undertaken to evaluate genetic diversity and the relationships between the three camel populations reared in Egypt: Maghrabi, Sudani, and Baladi using mitochondrial 16S sequences and other breeds of camels in the world. METHODS: Blood samples were collected from camels belonging to these three populations. Genomic DNA was extracted from the collected blood samples and subjected to PCR using specific primers for mitochondrial 16S region. The amplified products were purified using DNA purification kit to remove residual primers and dNTPs. Sequencing was performed in the Macrogen Incorporation. The amplified products were submitted to GenBank/NCBI under accession numbers OM 278349 and OM 278350 RESULTS: Sequencing was done on the partial mitochondrial 16S amplified fragments at 530 bp. This amplified area had two haplotypes. There was one substitution (G/A) at nucleotide 309 of the amplified segment. The nucleotide (π) and Hd stand for haplotype diversity, respectively, at 0.00008 and 0.042, and the average number of pairwise nucleotide differences, k, is 0.042, according to Fu's Fs statistic and Tajima's D, which is -1.10686. Genetic distance percentages between the three populations under study range from 0.000 to 0.0312. A phylogenetic analysis of Egyptian camel populations and other Camelus dromedarius populations revealed a strong relationship between them. CONCLUSIONS: This study suggests that the 16S rRNA sequencing in mitochondria plays a critical role in genetic variation studies and analysis of phylogeny between camel populations and breeds.

Five BoLA-DRB3 genotypes detected in Egyptian buffalo infected with Foot and Mouth disease virus serotype O.

Foot and Mouth disease (FMD) is a contagious disease leads to economically loss in livestock production all over the world. This serious disease is caused due to the infection of the animal with a single-stranded RNA virus (FMDV). This study aimed to investigate the genetic polymorphism of BoLA-DRB3 gene in Egyptian buffalo as a candidate genetic marker included in multi-factorial process of FMD resistance/susceptibility. Also this work aimed to genetically characterization and serotyping of circulating FMD virus in Egypt during 2016. For serotyping of FMDV, RT-PCR was used for FMDV-positive samples and the results declared the presence of serotype O in all tested animals. The sequence analysis of FMDV samples revealed five different patterns for the detected serotype O which were submitted to GenBank under the accession Nos.: MG017361-MG017365. The 302-bp amplified fragments from BoLA-DRB3 exon 2 were digested with HaeIII endonuclease and the results showed that the presence of five BoLA-DRB3 genotypes, among them the genotype AA might be associated with FMD-resistance (P < 0.01). On the other hand, genotype AC could be correlated with susceptibility (P < 0.01) to FMD in Egyptian buffaloes where it was absent in resistant group. The five detected genotypes of BoLA-DRB3 exon 2 were submitted to GenBank with the accession Nos.: MF977316-MF977320. In conclusion, our findings suggested that the detection of different BoLA-DRB3 genotypes may be has a promising role for raising the resistance of Egyptian buffalo against FMDV especially serotype O which is prevalent in Egypt with preferring genotype AA.

PCR-RFLP of bone morphogenetic protein 15 (BMP15/FecX) gene as a candidate for prolificacy in sheep.

Bone morphogenetic protein 15 (BMP15/FecX) gene is considered one of the major genes and a candidate marker for the reproduction in farm animals, especially sheep. The present study aimed to detect the genetic polymorphisms of BMP15 gene in sheep using PCR-RFLP technique. In the present study, 115 ewes were assigned into high and low prolificacy categories according to their reproductive history. In high prolific group (n = 20), ewes produced twins more than single births. In the low prolific type (n = 95), the ewes produced single births more than twins. DNA was extracted from blood samples of all ewes, subjected to PCR-RFLP analysis and confirmed by sequence analysis. The PCR products of 356 bp size were cut with HinƒI restriction enzyme. Three digested fragments of 70, 117 and 169 bp were obtained in both types of sheep. All animals were homozygous with CC genotype. In conclusion, the accessible findings did not detect any mutation in FecX gene in sheep, regardless their prolificacy. Therefore, further attempts are necessary to detect other SNP for BMP-15 gene in Egyptian sheep breeds.

Mitochondrial DNA genetic variations among four horse populations in Egypt.

Horses are one of the early domesticated animals in the world that changed societies and civilizations on a continent-wide scale. Due to the rare information about the genetic characterization of different horse populations in Egypt, this study aimed to identify the genetic biodiversity and relationships between four horse populations reared in Egypt. Genomic DNA was extracted and mtDNA region was amplified using polymerase chain reaction (PCR). The alignment of 384-bp amplified fragments showed the presence of 41 polymorphic sites resulting in 29 haplotypes which their sequences were submitted to GenBank under the accession numbers: KX909898-KX909926. The phylogeny tree for tested horses declared the presence of mixing maternal lineages between the four tested populations but still there are some separated lineages especially for Arabian and Thoroughbred horses. The sequences of 72 tested sequences were aligned with 13 published sequences as references, 11 of them for different Equus caballus whereas the other two reference sequences for Equus burchellii and Equus asinus. The results showed that all tested horses from the four populations are grouped with reference sequences of Equus caballus and separated from the other two reference sequences of Equus burchellii and Equus asinus. It is concluded that sequence analysis of mtDNA control region is still the most informative tool for the identification of genetic biodiversity and phylogeny of different horse breeds and populations. The horse populations reared in Egypt possess low genetic diversity and all of them are belonged to Equus caballus breed.

Cytochrome b conservation between six camel breeds reared in Egypt.

This study was aimed to assess cytochrome b conservation in six breeds of camels reared in Egypt and to compare its sequence with those of other livestock species. The 208-bp fragments from camel mtDNA cyto b were amplified using PCR for 54 camels belonging to 6 camel breeds reared in Egypt. The alignment of camel cyto b sequences showed the presence of two polymorphic sites resulting in four haplotypes and their nucleotide sequences were submitted to GenBank under the accession numbers: KX909894-KX909897. The genetic distances between tested camel breeds were zero between Baladi, Fallahi and Maghrabi breeds whereas they were at low value between the other three breeds: Mowaled, Sodany and Somali. Neighbor-joining showed 4 branches; one of them include most of the tested animals and another one contains 2 Somali animals which is considered a specific haplotype for this breed. The other two branches are mixed between Sodani and Mowaled breeds. Neighbor-joining tree was constructed between cyto b sequences of our tested camels and their sequences from livestock species include Camelus dromedaries, Camelus bactrianus, Ovis aries, Capra hircus, Bubalus bubalis, Bos Taurus and Sus scrofa. The result confirmed that our camel breeds belong to Camelus dromedaries and are clearly separated from other species. It is concluded that cyto b sequence is highly conserved among all camel breeds reared in Egypt which belong to Camelus dromedaries in addition to the advantage of cyto b in differentiation between different livestock sources which enables it to widely use for the adulteration detection in mixed meat.

Genetic characterization of Egyptian and Italian sheep breeds using mitochondrial DNA.

A 721-bp fragment from 15,541 to 16,261 bp (NC_001941.1) of the mtDNA control region from different Egyptian and Italian sheep breeds was amplified. The PCR products were purified and sequenced. From the amplified fragment of 721-bp, a region of 423 bp after excluding a central region rich in tandem repeats was analyzed. Within all tested breeds, the haplotype diversity and average number of pairwise differences were 0.97571 and 7.01484, respectively. The genetic distances (D) and the average number of pairwise differences (Dxy) between breeds were estimated. The lowest distance was observed between Laticauda and Italian Muflon followed by distance between Sarda and Italian Muflon while the highest distance was observed between Barki and Sarda followed by distance between Barki and Laticauda. Phylogenetic analysis showed the presence of three haplogroups - HapA, HapB and HapC - in the examined samples with the absence of other two haplogroups HapD and HapE. All Italian samples cluster with B haplogroup and also in the Egyptian breeds the most dominant haplogroup was B (62 out of 67 analyzed samples). In Egyptian Barki breed one individual clusters with A haplogroup and another individual with C haplogroup. In Ossimi breed two individuals cluster with C haplogroup and in Rahmani there is one sample belonging to A haplogroup. The matrix of pairwise differences among breeds was used to perform a Principal Component Analysis (PCA). This analysis showed that the Italian breeds are clearly separated from the Egyptian breeds; moreover the Egyptian Barki breed is separated from Ossimi and Rahmani.

Clastogenic effects of the fasciolicide drug fasinex on river buffalo lymphocyte cultures in vitro.

Fasinex (triclabendazole) has been reported to be an active fasciolocidal agent used in humans and in farm animals. The clastogenic effects of fasinex were tested in lymphocyte cultures of the river buffalo at three final concentrations: 25, 50 and 100 microg/ml. Chromosomal aberrations, sister chromatid exchanges and micronucleus formation are the three cytogenetic parameters used in this study. The results demonstrated that the number of cells with different types of chromosomal aberrations, including chromatid breaks and gaps, isochromatid breaks and gaps and polyploidy, was increased significantly in cultures treated with different doses of fasinex compared to the control. This increase was dose-dependent where there was a positive correlation between increased drug concentration and induction of chromosomal aberrations. The frequency of sister chromatid exchanges and the formation of micronuclei in all lymphocyte cultures treated with different doses of fasinex were increased significantly compared to the control; these increases were also dose-dependent. In conclusion, the three cytogenetic parameters used to evaluate the effect of fasinex revealed that the drug has a strong clastogenic effect on river buffalo lymphocytes in vitro.

Study on Genetic Polymorphism of IGFBP-3 Gene in Egyptian Buffalo

Aim: The present work was carried out to study the genetic characteristics of the IGFBP-3 gene in buffaloes reared in Egypt, where it is considered as one of the important molecular markers for productivity traits like growth and immunity in livestock species. One-hundred animals were used in this research work. Methods: The studied gene was amplified through polymerase chain reaction technique. Afterwards, the amplified fragment at 651-bp was digested with three different endonucleases; HaeIII, MspI, and TaqI. The genetic character of the IGFBP-3 gene was studied by using PCR-RFLP and nucleotide sequencing. Results: The PCR products after the digestion with those restriction enzymes revealed that the presence of the following fragments: two fragments at 506- and 145-bp with MspI two fragments at 240- and 411-bp with TaqI; and eight fragments at 199-, 164-, 154-, 56-, 36-, 18-, 16- and 8-bp with HaeIII. The restriction digestion of the amplified fragments of the IGFBP-3 gene did not show a genetic polymorphism or nucleotide substitution where all restricted fragments yielded from the digestion with three restriction enzymes were of the same sizes. Conclusion: Our findings indicated that the absence of the genetic polymorphism of the IGFBP-3 gene in Egyptian buffalo. Based on our results in addition to the significant effect of this gene on different productivity traits, the crossing between Egyptian buffalo with other breeds, particularly the Italian breed, is needed for more improvements of Egyptian buffalo's productivity where the Italian buffaloes characterized by high growth and fertility phenomena.

Genetic Structure and Nucleotide Sequences of Three Productivity Trait Genes, in Egyptian Buffalo

One of the main sources of meat and milk in Egypt is buffalo, of river type. The marker assisted selection-depending on the promised genetic markers is considered to be the effective way for improvement of buffalo productivity. This work aimed to study the genetic structure and nucleotide sequences of three productivity genes namely; LGB, PIT-1, and POUF-1 in Egyptian buffalo. This study is performed by using genomic DNA, which was extracted from 100 female buffaloes. The DNA extracts were subjected to PCR by using some specific primers of the tested genes. The PCR products were digested with dedicated restriction enzymes like; HaeIII for LGB and HinfI for both PIT-1 and POUF-1 genes. Depending on the appearance of restriction sites in the amplified fragments; GG

Cytochrome b Diversity and Phylogeny of Six Egyptian Sheep Breeds

Aim: Cytochrome b (Cyt-b) regions of mtDNA have received more attention due to their role in the genetic diversity and phylogenetic studies in different livestock. By using Cytochrome b sequencing, we aimed to clarify the genetic affinities and phylogeny of six Egyptian sheep breeds. Methodology: The genomic DNA was extracted and the specific primers were used for conventional PCR amplification of the Cytochrome b region of mtDNA (1134-bp) in sheep. The alignment of sequences was done to identify the sequence variations and polymorphic sites in the amplified fragments. Results: The results showed the presence of 39 polymorphic sites leading to the formation of 29 haplotypes (accession numbers: MG407500 - MG407528) with total haplotype diversity 0.814 and nucleotide diversity 0.00359. The lowest genetic distance was observed between Rahmani and Saidi while the highest distance was observed between Ossimi and Sohagi. The sequences of 111 analyzed samples were aligned with reference sequences of different haplogroups; A, B, C, D and E. The result showed that 86 out of 111 tested animals cluster with haplogroup B (77.48%), whereas 12 tested animals cluster with each of both haplogroups A and C (10.81%) and one animal belongs to haplogroup E (0.90%) with the absence of haplogroup D. Conclusion: Cytochrome b regions of mtDNA have an important role in the genetic diversity and phylogenetic studies in farm animals as well as many other mammalian species.

Genotyping and Nucleotide Sequences of Growth Hormone Releasing Hormone and Its Receptor Genes in Egyptian Buffalo.

Aim: The hypothalamic hormone, growth hormone-releasing hormone, is the principal stimulator of pituitary growth hormone (GH) synthesis and secretion. GHRH and its receptor (GHRHR) provide important functions in the regulation of the GH axis and in the development and proliferation of pituitary somatotropic axis. This study aimed to identify the genotypes and nucleotide sequences of two multifunctional genes; growth hormone-releasing hormone (GHRH) and its receptor (GHRHR) in Egyptian buffalo. Methodology: Genomic DNA was extracted from blood samples of 100 healthy buffaloes maintained at the Mahlet Mussa and El-Gmeasa herds from 2010 to 2012. PCR was performed using primers flanking a 296-bp fragment from GHRH gene and a 425-bp fragment from GHRHR gene of Egyptian buffalo. The PCR-amplified fragments were digested with HaeIII (GHRH) and Eco57I (GHRHR), electrophoresed and analyzed on agarose gels stained with ethidium bromide. The two amplified fragments were also sequenced and aligned with published sequences. Results: Depending on the presence of the restriction site at 241

Genetic Characterization of Insulin Growth Factor-1 and Its Receptor Genes in Egyptian Buffalo (Bubalus bubalis L.).

Aim: The somatotropic axis (SA) comprises genes associated with economically important quantitative traits in livestock like mammary and muscle growth as well as carcass traits. Insulin growth factor-1 (IGF-1) and its receptor (IGF-1R) are two important genes belonging to the SA. The aim of this study was to evaluate the genetic polymorphism of IGF1/SnaBI and IGF-1R/TaqI restriction sites in Egyptian buffalo. Methodology: Genomic DNA was extracted from blood samples of 100 healthy buffaloes maintained at the Mahlet Mussa and El-Gmeasa herds from 2010 to 2012. PCR was performed using primers flanking a 250-bp fragment of the regulatory region of the buffalo IGF-1 gene and a 616-bp fragment of the IGF-1R gene encompassing 51-bp from exon 12, 479-bp from intron 12 and 86-bp from exon 13. The PCR-amplified fragments were digested with SnaBI (IGF-1) and TaqI (IGF-1R), electrophoresed and analyzed on agarose gels stained with ethidium bromide. The two amplified fragments were also sequenced and aligned with published sequences. Results: All buffaloes investigated in this study were genotyped BB (i.e., negative for the SnaBI restriction site at position 224