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

Genotype of Melatonin Receptor MT1 (MTNR1A) and Puberty in Mediterranean Italian Buffalo

In adult buffaloes, polymorphism of the MT1 receptor gene has shown to influence the reproductive seasonality. The aim of study was to assess whether the polymorphism of the MTNR1A gene may influence puberty in Mediterranean Italian buffalo. The study was conducted using 50 prepubertal buffalo cows that at the age of 15 months were placed into the group where there was the male. Estrus detection was performed by observing estrous-behaviour and pregnancy checking by palpation per rectum and/or ultrasound between days 40 and 60 post-mating. Also of each animal dates of calving was recorded. From each buffalos a blood sample was collected and used for DNA extraction. PCR analysis was performed using 100-150 ng of DNA to amplify the second exon of the MTNRA1 gene. All PCR products were digested with 2U of enzyme HpaI to highlight the polymorphism at position 82 (characterized by a C to a T substitution) of the MTNR1A gene. Frequency of C and T alleles was respectively 0.42 and 0.58 in the analyzed population which resulted in Hardy Weinberg equilibrium. The genotypic frequency was 28% for genotype C/C, 38% for C/T and 34% for T/T. The registration of reproductive data showed that the first heat is around the age of 20 months and the first calving around 32 months. Our data show that the genotype of the MTNR1A does not influence the onset of reproductive activity in prepubertal buffalo cows

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

Causal link of total locomotor activity, melatonin and rectal temperature daily rhythm in small ruminants

To improve the knowledge in chronophysiology we investigated the causal link between the most important physiological variable studied until now; ten Sarda ewes and ten Sarda goats, pluriparus not pregnant and no lactating, were used. Animals were housed under natural environmental conditions in a common stall, alfalfa hay and water were available ad libitum. Each animal was equipped with an Actiwatch-Mini\uae for recording total activity. Blood samples were collected every 4 h over a 48 h period for the assessment of melatonin concentration. Rectal temperature was recorded with a digital thermometer immediately before the blood sampling at each data point. Single cosinor method showed a daily rhythm of studied variables. Higher MESOR and amplitude values of melatonin and rectal temperature were observed in sheep than in goats. The diurnal acrophase of locomotor activity was statistically different from the nocturnal acrophase of melatonin and rectal temperature, with no differences between the two species. Robustness was statistically lower in total locomotor activity in comparison with the others two variables, with a differences due to species in melatonin daily rhythm. In conclusion, in small ruminants, melatonin and rectal temperature daily rhythms are strictly correlated, and are not associated with the locomotor activity rhythm