We extracted a granule-rich sediment from normal human neutrophils and subjected it to chromatographic, electrophoretic, and functional analysis. The extract contained three small (molecular weight less than 3,500) antibiotic peptides that were named human neutrophil peptide (HNP)-1, HNP-2, and HNP-3, and which will be referred to as "defensins." HNP 1-3, a mixture of the three defensins, killed Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli effectively in vitro when tested in 10 mM phosphate buffer containing certain nutrients, but it had little or no bactericidal activity in nutrient-free buffer. In contrast, the nutrient-free buffer supported a high degree of activity by HNP 1-3 against Cryptococcus neoformans. In addition to its antibacterial and antifungal properties, HNP 1-3 directly inactivated herpes simplex virus, Type 1. Two of the individual purified defensins, HNP-1 and HNP-2, were as microbicidal as the mixture HNP 1-3. HNP-3 was less active than the other defensins against most but not all of the microbes tested. Immunoperoxidase stains revealed HNP 1-3 to have a granular localization in the neutrophil's cytoplasm by light microscopy. Frozen thin section immunogold transmission electron microscopy showed HNP 1-3 to be localized in azurophil granules. These studies define a broad-spectrum antimicrobial system in human neutrophils. The defensin system may operate in conjunction with or independently from oxygen-dependent microbicidal processes to enable human neutrophils to inactivate and destroy potential pathogens.

Defensins: natural peptide antibiotics in human neutrophils

Sheep milk has a high nutritional value and high concentrations of proteins, fats, minerals, and vitamins, as compared to the milks of other domestic species. The physicochemical and nutritional characteristics of sheep milk can be advantageous for the manufacture of products containing prebiotic ingredients and/or probiotic bacteria, which are major categories in the functional food market. Following this technological trend, this review will address the characteristics and advantages of sheep milk as a potentially functional food, as well as the development of sheep milk dairy products containing prebiotics and/or probiotics.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/1541-4337.12250

Sheep Milk: Physicochemical Characteristics and Relevance for Functional Food Development

Genome research in animals used in agriculture has progressed rapidly in recent years, moving from rudimentary genome maps to trait maps to gene discovery. These advances are the result of animal genome projects following closely in the footsteps of the Human Genome Project, which has opened the door to genome research in farm animals. In return, genome research in livestock species is contributing to our understanding of chromosome evolution and to informing the human genome. Enhancement of these contributions plus the much anticipated application of DNA-based tools to animal health and production can be expected as livestock genomics enters its sequencing era.

http://webpages.icav.up.pt/PTDC/CVT/099782/2008/References%20for%20Exomics%20proposal%20%5BPTDC-CVT-099782-2008%5D/PDF%20files%20of%20references_max%2030%20%5BExomics%5D/Womack%202005%20GenomeRes_Advances%20in%20livestock%20genomics-Opening%20the%20barn%20door.pdf

Advances in livestock genomics: opening the barn door.

Asian buffaloes (Bubalus bubalis) have an important socio-economic role. The majority of the population is situated in developing countries. Due to the scarce resources in these countries, very few species-specific biotechnology tools exist and a lot of cattle-derived technologies are applied to buffaloes. However, the application of cattle genomic tools to buffaloes is not straightforward and, as results suggested, despite genome sequences similarity the genetic polymorphisms are different. The first SNP chip genotyping platform designed specifically for buffaloes has recently become available. Herein, a genome-wide association study (GWAS) and gene network analysis carried out in buffaloes is presented. Target phenotypes were six milk production and four reproductive traits. GWAS identified SNP with significant associations and suggested candidate genes that were specific to each trait and also genes with pleiotropic effect, associated to multiple traits. Network predictions of interactions between these candidate genes may guide further molecular analyses in search of disruptive mutations, help select genes for functional experiments and evidence metabolism differences in comparison to cattle. The cattle SNP chip does not offer an optimal coverage of buffalo genome, thereafter the development of new buffalo-specific genetic technologies is warranted. An annotated reference genome would greatly facilitate genetic research, with potential impact to buffalo-based dairy production.

/pdf/prospecting-major-genes-in-dairy-buffaloes-26c2uejr65.pdf

Prospecting major genes in dairy buffaloes

Sheep is the second most important dairy species after cow worldwide, and especially in the Mediterranean and Middle East regions. In some countries, the difficult environmental conditions require a peculiar adaptation and, in these contexts, sheep are able to provide higher quality protein than cattle. In the least-developed countries, the amount of dairy sheep and ovine milk production is progressively increasing. In order to improve dairy productions, in particular those with local connotations, it is necessary to obtain in-depth information regarding milk quality and rheological properties. The genetic polymorphisms of milk proteins are often associated with quantitative and qualitative parameters in milk and are potential candidate markers that should be included in breeding strategies similar to those already available for cattle. Due to the current and growing interest in this topic and considering the large amount of new information, the aim of this study was to review the literature on sheep milk protein polymorphisms with a particular emphasis on recent findings in order to give scientists useful support. Moreover, the effects of different protein variants on milk yield and composition are discussed. © 2014 Society of Chemical Industry

Investigating the genetic polymorphism of sheep milk proteins: a useful tool for dairy production.

Genetic variation in three Egyptian indigenous sheep breeds namely: Barki, Ossimi and Rahmani were investigated using fourteen microsatellite loci. The total number of alleles ranged from 6 in CSSM47 locus to 14 in TGLA 377 locus. The fourteen tested loci were all polymorphic in the three breeds. Major differences between the breeds were found at ten of the tested loci, where the alleles at the highest frequency are different in the three breeds. While, at loci OARCP20, OARVH72, CSSM47 and OARAE129, two of the tested breeds have similar alleles at the highest allele frequency. The average direct count of heterozygosity overall loci in each tested breed was less than the expected heterozygosity. Tests of genotype frequencies for deviation from the Hardy-Weinberg equilibrium (HWE), at each locus overall breeds, revealed significant departure from HWE due to heterozygote deficiency. A slightly high rate of inbreeding within the three breeds was noticed (global FIS = 0.308). Low genetic differentiation was detected by estimation of FST index between all pairs of breeds. Cluster analysis revealed that Ossimi and Rahmani breeds clustered independently from Barki breed at 0.43 of genetic distance. The obtained results can be useful for the development of a rational breeding strategy for genetic improvement of sheep in Egypt.

/pdf/analysis-of-genetic-variation-in-different-sheep-breeds-5f8xwadvfg.pdf

Analysis of genetic variation in different sheep breeds using microsatellites

The cosegregation of ten coding loci has been investigated, in a panel of 37 somatic cell hybrids resulting from the fusion of a hamster cell line and river buffalo lymphocytes, by use of Southern hybridization technique. Five syntenic groups, TCRB-PGY3, ASS-ABL, FUCA1P-CRYG, MBP-YES1, and CGN1-ACTA1, previously assigned to cattle as U13, U16, U17, U28, and U29 respectively, were also found to be syntenic in buffalo. Based on the extensive syntenic conservation and banding homology between cattle and river buffalo, comparative mapping predicts the localization of these syntenic groups on river buffalo Chromosomes (Chrs) :BBU7, BBU12, BBU2q, BBU22, and BBU4q respectively as they have been previously localized on cattle Chrs BTA4, BTA11, BTA2, BTA24 & BTA28.

Synteny mapping in river buffalo.

2 Abstract: Sheep is one o f the m ost important domestic ani mals in E gypt like m any other developing countries. It is ra ised mainly for m eat p roduction with carpet she ep wool as a secondary p roduct. Using RAPD-PCR assay, t he genetic variation among five Egyptian sheep breeds n amely: Rahmani, Ossimi, Barki, Saidi and Soh agi were analyzed. Fifty-seven bands were amplified using five random primers with a mean of 11.4 ba nds per primer. The mean number of shared bands between all breeds ranged from 5 t o 11 with a mean value o f 8.0. Fifty six out of 57 total amplified fragments (98.25%) ar e polymorphic loci, useful for genet ic vari ation s tudy between breeds. The genet ic si milarity between five Egyptian sheep breeds ra nged from 0.8332 to 0.9750. S ix b ands w ere found to be useful as sp ecific makers for the studied sheep breeds. UPGMA dendrogram grouped the investigated sheep breed genotypes into two main c lusters. T he first clu ster in cludes O ssimi and Rahmani, whereas the second is divided into two sub-clusters: O ne includes B arki an d the other in cludes S aidi an d Sohagi.

Genetic Variation Between Some Egyptian Sheep Breeds Using RAPD-PCR

The major whey proteins, β-lactoglobulin (β-LG) and α-lactalbumin (α-LA), are strongly correlated with the nutritional value and the functional properties of milk whey. Ovine whey has a unique protein composition compared to that of bovine whey and it is especially rich in proteins. Genetic polymorphism of β-LG using PCR-RFLP and α-LA using PCR-SSCP was analyzed in three native Egyptian sheep breeds named Barki, Rahmani and Ossimi. Digestion of resulting amplified fragments of β-LG (452-bp) with RsaI endonuclease differentiated between three different genotypes AA (69.77%), AB (26.74%) and BB (3.49%). The results showed that A allele was present in the Egyptian sheep breeds with a frequency (83.14%) higher than B allele (16.86%). A & B alleles appeared at frequencies 70.4 % & 29.6%, 95.0% & 5.0% and 82.8% & 17.2% in Barki, Rahmani and Ossimi, respectively SSCP results of the α-LA amplified fragments (166-bp) showed that the three native Egyptian sheep breeds possess two different patterns related to the difference in nucleotide sequences with frequencies 88.1% and 11.9%. The frequencies of pattern 1 ranged from 81.48% in Barki to 89.66% in Ossimi and 92.86% in Rahmani breeds whereas the pattern 2 was present at frequencies 18.52%, 7.14% and 10.34% in Barki, Rahmani and Ossimi breeds, respectively

Genetic Polymorphism of Whey Protein Genes β-LG and α-LA in Three Egyptian Sheep Breeds

Objective: Casein genetic polymorphisms are important and well known due to their effects on quantitative traits and technological properties of milk manufacturing. The casein fraction of ruminant milk proteins consists of four caseins, namely 8s1-, 8s2-, &#946-and K-casein. At the DNA level, polymerase chain reaction (PCR) and single-strand conformation polymorphism (SSCP) allow for the simultaneous typing of several alleles at casein loci, as well as the detection of unknown polymorphisms. The genetic polymorphism of two ovine milk protein casein genes, &#946- and K-caseins, was studied in sheep animals belonging to three main breeds reared in Egypt (Rahmani, Barki and Ossimi), as a tool for genetic improvement of milk trait characteristics.
Methodology and results: SSCP of &#946-casein exon 7 revealed two different patterns in eighty-five tested animals. The sequence analysis of the PCR product (299-bp) of these two different patterns showed two single nucleotide substitutions; A®C and C®T without any amino acid exchange. The frequencies of these two different patterns were 96.67% and 3.33% in Rahmani; 65.52% and 34.48% in Ossimi and 88.46% and 11.54% in Barki, respectively. The nucleotide sequence of 9-casein in Egyptian sheep was submitted in database NCBI/ GenBank with the accession number JX080379. The polymorphism of Kcasein gene was also detected in eighty-six animals using PCR-SSCP technique. PCR amplified a
fragment with 406-bp in exon 4 of this gene. SSCP results showed that all tested sheep animals are monomorphic. The alignment between our sequences with published sequence revealed two nucleotide substitutions; C®T and T®C. The nucleotide sequence of K-casein in our tested animals was submitted in database NCBI/ GenBank with the accession number JX050176.
Conclusion and application of findings: This study aimed to identify the genetic polymorphism of &#946 and Kcasein genes, which are strongly related to economically important milk quantitative traits in some Egyptian sheep breeds, as a tool or genetic markers for improvement of these breeds.

 Key words: Sheep, &#946-casein, K-casein, PCR-SSCP, SNP.

/pdf/genetic-variations-of-946-and-k-casein-genes-in-egyptian-rhbdzcm4pt.pdf

Eman R. Mahfouz

Papers

Analysis of genetic variation in different sheep breeds using microsatellites

Synteny mapping in river buffalo.

Genetic Variation Between Some Egyptian Sheep Breeds Using RAPD-PCR

Genetic Polymorphism of Whey Protein Genes β-LG and α-LA in Three Egyptian Sheep Breeds

Genetic variations of &#946- and K-casein genes in Egyptian sheep breeds