PCR-SSCP analysis of GH gene in Sarda goats: a high variability and its preliminary effects on dairy performances

The growth hormone (GH) gene can be utilized as a major gene because in various domestic livestock its polymorphisms have been associated to milk traits. The aim of this research was to investigate single-strand conformation polymorphism (SSCP) in the exon 3 of gGH (goat GH) gene and to evaluate the possible association with milk traits in Sarda goat breed. Forty-four primiparous lactating goats were randomly chosen, and the productive parameters (milk yield, fat, protein, and lactose percentage) of three consecutive lactations were monitored. The exon 3 of the gGH gene was PCR amplified and the resulting products were analysed by SSCP. Six conformational patterns were detected. The sequencing of SSCP patterns revealed the occurrence of six nucleotide changes, two of which determined amino acid changes in the deduced protein sequence. A preliminary comparative analysis of the productive traits related to three lactations with the genomic profiles derived from the SSCP analysis was performed with the ANOVA statistical method. SSCP polymorphic patterns in exon 3 were associated (P<0.01) with milk yield, fat and protein percentages, and with lactose content (P<0.05). These findings may be used for marker assisted selection in Sarda goat, in order to improve dairy production, preserving genetic diversity of the population

Testicular degeneration and infertility following arbovirus infection

Arboviruses can cause a variety of clinical signs including febrile illness, arthritis, encephalitis and hemorrhagic fever. The recent Zika epidemic highlighted the possibility that arboviruses may also negatively affect the male reproductive tract. In this study, we focused on bluetongue virus (BTV), the causative agent of bluetongue and one of the major arboviruses of ruminants. We show that rams that recovered from bluetongue displayed signs of testicular degeneration and azoospermia up to 100 days after the initial infection. Importantly, testicular degeneration was induced in rams experimentally infected with either a high (BTV-1IT2006) or low (BTV-1IT2013) virulence strain of BTV. Rams infected with the low virulent BTV strain displayed testicular lesions in the absence of other major clinical signs. Testicular lesions in BTV-infected rams were due to viral replication in the endothelial cells of the peritubular areas of the testes, resulting in stimulation of a type-I IFN response, reduction of testosterone biosynthesis by Leydig cells, and destruction of Sertoli cells and the blood-testis barrier in more severe cases. Hence, BTV induces testicular degeneration and disruption of spermatogenesis by replicating solely in the endothelial cells of the peritubular areas unlike other gonadotropic viruses. This study shows that a naturally occurring arboviral disease can cause testicular degeneration and affect male fertility at least temporarily

Relationships between milk characteristics and somatic cell score in milk from primiparous browsing goats

To determine milk yield and composition, total microbic count (TMC) and somatic cell count (SCC) of browsing goats throughout the first lactation, 100 goats of Sarda breed, equally distributed in four flocks (F1, F2, F3 and F4), were selected. They were exclusively fed pasture and hand-milked once daily. Individual milk samples and daily milk yield were taken from each goat at monthly intervals, from March to July. Milk samples were analyzed for: total protein, fat, lactose, urea, freezing point (FP), pH, TMC and SCC. The data was subjected to analysis of variance and to correlation matrix. On the whole, in all the flocks, milk yield showed the highest production in April and May. Fat content increased (P &lt; 0.01) throughout the lactation. Protein content showed the lowest value (P &lt; 0.01) in June (4.15%). Urea and pH values were fluctuating. FP was lower (P &lt; 0.01) at the start of lactation (−0.562 Hortvet degrees). TMC log10 values were low, considering the hand milking and inadequacy of facilities on the farms. SCC increased (P &lt; 0.01) throughout the lactation and, on the whole, SCC and TMC were not correlated

A Reliable method for characterization of the goat <i>CSN1S1</i> E allele

Goat αs1-casein, coded by the CSN1S1 gene, is a highly polymorphic protein. The E allele is characterized by the insertion of a long interspersed nuclear element (LINE) in the 19th exon of the gene. This insertion is probably responsible for the reduced levels of αs1-casein found in milk from animals expressing the E allele. The present study aimed to develop a robust method for the detection of the CSN1S1 E allele in goat genomic DNA, using a one-step allele-specific polymerase chain reaction (AS-PCR). Three primers were designed, based on published DNA sequences (GenBank: AJ504710 and X72221). The primers were used simultaneously in the amplification reaction. Genomic DNA samples from animals with known CSN1S1 AA, EE and AE genotypes were used as positive controls. The DNA fragments were analysed by agarose gel electrophoresis, and were found to be of the expected sizes. The fragment characterizing the A allele (A refers to the αs1-casein alleles bearing an intact 19th exon) was 583 bp long, and the fragment characterizing the E allele was 437 bp long, spanning part of the 18th intron, part of the 19th exon and 146 bp of the LINE inserted sequence. This method allows clear identification of the three genotypes of CSN1S1 (A*A*, A*E and EE) using a one-step PCR. Using this method, the frequency of the CSN1S1 E allele in a population of 300 Sarda goats was estimated to be 0.037

Sequence of cDNA coding for a 65 kDa adhesive protein for the specific detection of Trichomonas vaginalis by PCR

A Trichomonas vaginalis cDNA library was constructed and recombinant plaques were screened using rabbit immunoglobulins specific for P65, a protozoan protein involved in pathogenicity that we identified in a previous study. A 1.38 kilobases cDNA fragment coding for the P65 protein was cloned in E. coli and then sequenced. On the basis of of the sequence obtained, six primers were synthesised and used to set up a Polymerase Chain Reaction. The presence of a specific amplicon in all 30 clinical isolates tested shows that P65 is a conserved and stable gene. The reaction is highly sensitive (as few as 5 to 10 parasites can be detected) and specific for Trichomonas vaginalis; the gene coding P65 adhesin can be therefore considered a very good molecular target for polymerase chain reaction-based diagnostic purposes

Epidemiologia molecolare degli SRLV nel mediterraneo.

Trabajo presentado en el VI Workshop di Virologia Veterinaria, celebrado en Turín (Italia), los días 13 y 14 de octubre de 2016Peer reviewe

Nutritional and sensory properties of mouflon x Sarda lamb meat

Suckling lamb is a typical foodstuff of the Mediterranean dairy sheep breeding. The milk lamb produced in Sardinia is much appreciated for its organoleptic characters. Nevertheless, since it is fed with ewe-milk, it has production costs that are not entirely repaid by market prices. A greater exploitation occurred since PGI (geographical indication) was accomplished, but an improvement could be attained also through crossbreeding between dairy and meat breeds. The aim of this research was to determine nutritional and sensory traits of the lamb meat obtained from the crossbred between Mouflon and Sarda sheep breed. Ten carcasses of the crossbred were compared to ten carcasses of pure Sarda breed. All the carcasses were classified for weight, quality, meat color and fat class. The acidic profile was determined on intramuscular and depot fat using gaschromatography. Sensorial investigation was performed on meat after cooking, using an affective method. Data were submitted to statistical analysis. Crossbred meat is leaner and it provides a lower caloric intake (P&lt;0.01). Fatty acid profile of fat depots were similar between the two groups, while as regards the composition of intramuscular fat, Sarda lambs had a higher percentage of MUFA (P&lt;0.01) and lower nutritional indexes. Consumer test indicated a clear preference (P&lt;0.01) for crossbred meat, that has a stronger taste, even though it is slightly less tender. On the whole, data pointed out a good nutritional quality for meat of both genotypes, but crossbred meat has better sensorial characteristics

Whole-Genome Sequencing of Two Canine Herpesvirus 1 (CaHV-1) Isolates and Clinicopathological Outcomes of Infection in French Bulldog Puppies

Canine herpesvirus 1 (CaHV-1) infects dogs, causing neonatal death and ocular, neurological, respiratory, and reproductive problems in adults. Although CaHV-1 is widespread in canine populations, only four studies have focused on the CaHV-1 whole genome. In such context, two CaHV-1 strains from both the kidney and spleen of 20-day-old deceased French Bulldog puppies were recently isolated in Sardinia, Italy. The extracted viral DNA underwent whole-genome sequencing using the Illumina MiSeq platform. The Italian CaHV-1 genomes were nearly identical (>99%), shared the same tree branch, and clustered near the ELAL-1 (MW353125) and BTU-1 (KX828242) strains, enlarging the completely separated clade discussed by Lewin et al., in 2020. This study aims to provide new insights on the evolution of the CaHV-1, based on high-resolution whole-genome phylogenetic analysis, and on its clinicopathological characterization during a fatal outbreak in puppies

A Deeper Insight into Evolutionary Patterns and Phylogenetic History of ORF Virus through the Whole Genome Sequencing of the First Italian Strains

Orf virus (ORFV) is distributed worldwide and is the causative agent of contagious ecthyma that mainly occurs in sheep and goats. This disease was reported for the first time at the end of 18th century in Europe but very little is currently known about the temporal and geographic origins of this virus. In the present study, the use of new Italian whole genomes allowed for better inference on the evolutionary history of ORFV. In accordance with previous studies, two genome types (S and G) were described for infection of sheep and goats, respectively. These two well-differentiated groups of genomes originated for evolutive convergence in the late 1800s in two different areas of the world (Europe for S type and Asia for G type), but it was only in the early 1900s that the effective size of ORFV increased among hosts and the virus spread across the whole European continent. The Italian strains which were sequenced in the present study were isolated on the Mediterranean island of Sardinian and showed to be exclusive to this geographic area. One of them is likely representative of the early European forms of ORFV which infected sheep and became extinct about one century ago. Such an ancient Sardinian strain may have reached the island simple by chance, where it quickly adapted to the new habitat