Genotype at the MTNR1A locus and response to melatonin treatment in Sarda lambs

With the aim to evaluate the effect of melatonin treatment and melatonin receptor 1A (MTNR1A) genotype on advance of puberty, 423 Sarda lambs were chosen. On June 26th, they were divided into three groups, each of 141 animals (groups 0, 1, and 2), on the basis of live weight. On June 30th, animals in group 1 received a single implant (18 mg melatonin), while group 2 received two implants. Group 0 was untreated. Thirty-five days after treatment (August 4th), rams were introduced and after 40 days they were removed. From January 1st to February 10th lambing dates were recorded. Genomic DNA was extracted and subjected to PCR for the amplification of exon II and then digested with enzymes MnlI and RsaI and placed into +/+, +/−, or −/− group for MnlI and C/C, C/T, or T/T group for RsaI. Samples were cloned and sequenced. Data obtained were subjected to χ2 test in order to evaluate the difference in fertility among groups and the link between genotype and reproductive activity. Genotype +/+ and C/C showed the highest incidence. Treated groups showed a higher number of lambing at 10th February compared to control group (P<0.04). Melatonin treatment results more efficient in +/+ genotype

Reproductive activity in sheep with different lambing period treated with melatonin in April

The object was to evaluate the effect of melatonin treatment on the advance in April of the reproductive resumption in Sarda breed sheep with different lambing period. For the research two farms, located in North Sardinia between 39° and 40° N, were chosen. In each farm, 120 lactating ewes were selected: 30 lambed between October 20th and November 20th (group 1); 30 lambed between December 1st and 30th (group 2); 30 lambed between January 1st and 30th; 30 lambed between February 1st and 28th (group 4). In each farm, each group of 30 animals was divided into two subgroups of 15 animals (M and C). On April 1st, in each farm, the animals of the M subgroups were treated with a implant containing 18 mg melatonin. The subgroups C were kept as control. The lambing dates and the number of newborn lambs were recorded until 220 days after ram introduction. In treated animals greatest fertility (P<0.01) and lowest distance in days from male introduction to lambing (P<0.01) were recorded. The best reproductive performances were found in the group 1 and 2 compared to the other two groups (P<0.01).The present research shows that melatonin treatment should be made 3 or 4 months after lambing, in order to obtain optimal results

Polymorphism of the MTNR1A Melatonin Receptor Gene in Goat Breeds of Northern Italy

Melatonin receptor MT1, encoded by the MTNR1A gene, is the main one involved in the seasonal regulation of reproductive activity. A correlation between this gene polymorphism and reproductive performance has been demonstrated in sheep. To date, no data about MTNR1A gene polymorphism are available regarding Italian goat breeds other than the Sarda goat. This study aimed to detect any PCR-RFLP polymorphic sites of MTNR1A using MnlI and RsaI enzymes in Northern Italian goat breeds, which are characterized by a pronounced reproductive seasonality. One-hundred-eight adult female goats belonging to four different breeds were included in the study (i.e., Frisa Valtellinese, n = 29; Orobica, n = 23; Lariana, n = 29; Camosciata delle Alpi, n = 27). Blood was sampled from each goat. Genomic DNA was extracted from each sample and the main part of exon II of MTNR1A gene was amplified by PCR and digested with MnlI and RsaI enzymes. Unexpectedly, none of the fragments were found to be polymorphic. The absence of polymorphism may be linked to the macro group of goat breeds that evolved during human migrations. Breeds of the Alpine-European strain would appear to show no polymorphism, as confirmed by our study, whereas breeds belonging to the Mediterranean-African or Asian-Middle Eastern strains do

Characterization of the SREBP-1 Gene Polymorphisms and Milk Traits in Dairy Sheep

The SREBP genes (Sterol Regulatory Element-Binding Proteins) are involved in the milk fat synthesis. In dairy cows some polymorphisms at the SREBP-1 gene sequence have been related with milk fat content. The aim of this study was to characterize the entire coding regions of the SREBP-1 gene in Sarda sheep breed, in order to highlight any polymorphisms and their association with milk traits. Four-hundred adult and lactating Sarda ewes were selected. Individual milk yield was recorded monthly from Day 30 to Day 150 of lactation, and fat and protein concentration were analysed. A blood sample from each ewe was taken for DNA extraction; thus, all the 19 coding exons of the SREBP-1 gene were amplified by polymerase chain reaction (PCR). Single-strand conformation polymorphism analysis (SSCP) and sequencing were used to scan mutations. Results provide, for the first time, the entire coding DNA sequence (CDS) of the SREBP-1 gene in sheep, and by sequences analysis 8 polymorphisms have been detected. The statistical analysis exhibited no relationship between polymorphisms and milk traits. The low SREBP-1 gene diversity that emerged from the present study, may be linked to the important role of this gene in the mechanism of milk fat synthesis or to the severe genetic selection performed in the Sarda sheep. However, it would be necessary to extend the study, including other breeds and other genes, in order to expanding the knowledge about the process of milk fat synthesis in dairy sheep

Reproductive Resumption in Winter and Spring Related to MTNR1A Gene Polymorphisms in Sarda Sheep

The aim of the present research was to evaluate the association between the SNPs rs430181568 and rs407388227 located in the MTNR1A gene with the reproductive recovery of Sarda sheep in different months of ram introduction in the flock (February, March, April and May). In order to address this research gap, we selected two farms, each of which consisted of approximately 1000 animals; a total of 800 ewes (400 for each farm) were genotyped for the two single nucleotide polymorphisms rs430181568 and rs407388227 located in the exon 2 of the MTNR1A. These SNPs are completely linked; thus, each genotype of rs430181568 corresponded to the same genotype for rs407388227. Among the genotyped animals, 240 individuals were selected and divided into four homogeneous groups (A, B, C and D) of 60 subjects, each group based on age (range 3&ndash;6 years old), body condition score (BCS) (range 2.0&ndash;4.0) and genotype (20 ewes carrying CC/CC, 20 CT/CT and 20 TT/TT genotype). The dates of the ram introduction in each group were 15 February, 15 March, 15 April and 15 May, respectively. In all groups, the lambing date and the number of lambs born from 150 to 220 days after the ram introduction were recorded. In all the groups, the genotypes CC/CC and CT/CT of the polymorphism (rs430181568 and rs407388227) showed the greatest fertility (the ratio between the number of lambed ewes and the ewes exposed to the rams) (p &lt; 0.01) and the shortest distance between ram introduction to lambing (p &lt; 0.01), compared with the TT/TT genotype. In conclusion, we determined that the polymorphisms rs430181568 and rs407388227 were associated with reproductive recovery, after ram introduction, even in February and March, months subsequent to the photorefractoriness period

Effect of genotype at the MTNR1A locus and melatonin treatment on first conception in Sarda ewe lambs

A sample of 423 Sarda ewe lambs from three different farms was used to evaluate the effect of one or two melatonin implants on the time of first conception. On each farm, 141 animals were divided into three groups. On June 30 these animals received either no treatment (Group C), 18 mg melatonin (Group M1, one implant), or 18 + 18 mg melatonin (Group M2, two implants). Thirty-five days after treatment, rams were introduced in the ewe lambs flock and subsequently removed after 40 days. Lambing dates were recorded between 150 and 190 days from the first day of male introduction. Genotyping and sequencing of the MT1 exon 2 were carried out to analyze the structure and the possible influence of the MT1 receptor gene on reproductive response to melatonin treatment. Melatonin-treated animals had a higher rate of pregnancy (P &lt; 0.05) and lambed earlier (P &lt; 0.05) compared with untreated animals. Single nucleotide polymorphisms were found in exon II of MT1 gene at positions C606T and G612A leading to genotypes C/C, C/T or T/T and +/+, +/− and −/−, respectively. Melatonin-treated animals of +/+ genotype showed a higher number of pregnancies (P &lt; 0.05) and lambed earlier (P &lt; 0.05) compared to untreated animals of the same genotype. Melatonin treatment did not affect reproductive activity in any other genotype analyzed. No correlation between genotype and the time of first conception was found in untreated animals. Concluding data revealed the positive effect of melatonin treatment on the time of first conception in ewe lambs and highlighted that +/+ genotype is able to influence reproductive response to melatonin treatment

A Polymorphism at the melatonin receptor 1A (<i>MTNR1A</i>) gene in Sarda ewes affects fertility after AI in the spring

The effect of MTNR1A gene polymorphisms on the fertility rate after AI in Sarda sheep was evaluated in 600 lactating adult ewes. Genomic DNA was subjected to amplification of the MTNR1A gene exon II. Amplicons were digested with restriction endonuclease MnlI. Ten samples from each genotype were sequenced. A polymorphism was detected (A612G) and ewes were determined to be +/+, +/– or –/– for the allele. Allelic frequency was 0.77 for the + allele and 0.23 for the – allele. The frequency of the +/+, +/– and –/– genotypes was 68, 19 and 13%, respectively. On 16 May 2009, 60 ewes from each genotype group were synchronised using intravaginal sponges containing 40 mg fluorogestone acetate for 14 days. At sponge removal, the ewes were administered 350 IU pregnant mare’s serum gonadotropin and were then inseminated, 54–56 h later, with 400 × 106 spermatozoa. Pregnancies were confirmed 50 days after AI using transabdominal ultrasonography. Lambing dates and the number of newborn lambs were recorded within 155 days after AI. Conception and lambing rate were higher for ewes with the +/+ and +/– genotypes compared with those with the –/– genotype (P &lt; 0.01). In conclusion, there was a positive correlation between MTNR1A allele polymorphisms the reproductive response following synchronisation and AI in the spring

Discrepancies between Genetic and Visual Coat Color Assignment in Sarcidano Horse

This study aimed to evaluate the discrepancies between genetic and visual coat color assignment in the Sarcidano Horse and to elucidate potential reasons. Individual DNA from 90 Sarcidano Horses was used for genetic assignment of coat color to explore the correspondence with individual forms containing phenotypical traits. The MC1R exon 1 and ASIP exon 3 have been genotyped and sequenced to obtain a picture of the coat color distribution in this breed. Surprisingly, once we compared the genetic results with the individual forms reporting the phenotypic data for each subject, a certain degree of non-correspondence between the phenotypic and genetic data in relation to coat color emerged. From the genetic analysis, Chestnuts (n = 58) resulted the most common Sarcidano Horse (n = 58), followed by a quite large number of Blacks (n = 28) and a very small number of Bays (n = 4), whereas phenotypic distribution resulted in 38 Chestnuts, 40 Bays, only 2 Blacks, and 10 Grays (without the possibility of recognizing the true color they carried). Chestnut resulted a very representative coat color, while many horses that visually identified as Bays were genetically Blacks. This discrepancy, that could be due to a variety of individual and external factors, including age, time of year, living situation and dietary condition, suggesting the importance of accurate coat color identification to ensure adequate features registration and reliable prediction of offspring’s coat color

Effects of melatonin treatment on milk traits, reproductive performance and immune response in Sarda dairy sheep

In ruminants, the role of melatonin in the control of reproductive seasonality is well reported, but it is still little known about its action on milk traits and on the immune system. The aim of this study was to assess the effect of melatonin on milk yield and composition, somatic cell count (SCC), some cytokine blood concentration and reproductive resumption in sheep. One hundred lactating sheep were allocated to two groups (of 50 sheep each), M (treated with melatonin) and C (controls), and exposed to the rams for 50 d. Time period in days from ram introduction to lambing (TRIL) and litter size were recorded. Every 15 d, from 1 March to 30 April, the individual daily milk yield was registered and milk composition and SCC, were analysed. The levels of Interleukin 2 and 6 (IL-2 and 6) and tumour necrosis factor-alpha (Tnf-α) were evaluated every 15 d. The highest fertility rate (p < .01) and the shortest TRIL (p < .05) was recorded in M group. Milk yield and composition were similar between groups. The somatic cells decreased continuously in the treated ewes (p < .01) and increased continuously in untreated ewes (p < .05). IL-2 and IL-6 increased in M group, although no statistical differences were found between groups. Melatonin administration improved reproductive efficiency, did not affect milk yield and composition and decreased somatic cells. This last effect could help to maintain healthy mammary gland and encourage farmers to reduce the use of drugs against mastitis.Highlights Melatonin is involved in different physiological processes, such as the control of reproductive seasonality, milk production and the immune response. Melatonin administration in Sarda sheep did not modify milk yield and composition, decreased SCC and advanced reproductive resumption in spring. Melatonin could be used to strengthen the sheep immune response in order to reduce the use of drugs for both therapeutic and preventive purposes against mastitis