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ABSTRACT: The aim of this research was to investigate variability in each of the five exons of the caprine growth hormone (gGH) gene, in order to establish the possible relationships with milk traits in Sarda breed goat. The general linear model procedure was used to analyse the effects of the single strand conformation (SSCP) profiles on milk traits of 100 lactating goats. Analysis of conformational polymorphism at exons 1-5 revealed a total of 25 differing banding patterns. Sequencing revealed 21 nucleotide changes (compared with GenBank D00476): 14 were polymorphic and 7 monomorphic; 19 in exonic regions, 5 of which were nonsynonymous. A SNP upstream of the transcription initiation codon (c.-3A>G) and an indel (c.*29_30insC) in the 3'UTR, were detected. Alignment of 4 cloned sequences including the entire gGH gene led to the identification of 22 nucleotide variations within the intron regions, including two indels. Association analysis revealed that each exon, except exon-1, affected milk yield, exons 1 and 3 influenced milk fat percentage, and all exons, except exon-2, had an effect on protein percentage, supporting previous results in livestock. The variability detected at the caprine GH gene might provide useful information for the phylogeny of ruminants and, more importantly, have implications on the biological function of the growth hormone and on those traits resulting from its physiological action, including milk production and composition. The caprine GH gene may become a useful molecular marker for a more effective genetic selection for milk production traits in goats.
Journal of Dairy Research 04/2013; · 1.34 Impact Factor
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ABSTRACT: The effect of CSN1S1 genotype and lactation stage on milk yield and composition were investigated in 80 extensively reared goats. Milk yield was recorded in early, mid and late lactation and individual milk samples were collected to determine: fat, protein, lactose and casein content, pH, freezing point, somatic cell count (SCC) and total microbic mesophilic count (TMC). Relative casein composition and amino acid profile were quantified by HPLC. Fatty acid profile was measured by gas-chromatography. Genotype did not affect milk yield, while this trait was significantly affected by lactation stage (P < 0·01). CSN1S1 BB goats produced significantly higher protein and casein percentages (P < 0·05). αs1-casein (CN) was significantly higher in BB and AB goats than AF and BF, showing intermediate values in AA goats (P < 0·01). The protein percentage and the αs1 and αs2-CN fractions were not affected by lactation stage, while the casein content and the β and κ-CN significantly increased throughout lactation (P < 0·01). C4 : 0 and C6 : 0 were not affected by genotype, while C8 : 0 and C10 : 0 were higher in the AA goats than BB; most of the long chain FA were higher in BB than AA goats. MUFA and PUFA increased in late lactation. In addition, BB goats showed higher essential amino acids, resulting in an optimal composition from the nutritional point of view, when compared with AA goats. The increase of MUFA, PUFA, essential and cis-FA in late lactation indicate that the lipid composition of goat's milk, with the progress of lactation, tends to improve its nutritional value.
Journal of Dairy Research 01/2013; · 1.34 Impact Factor
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ABSTRACT: The aim of the study was to detect polymorphism in the POU1F1 gene in Sarda breed goat, as well as to establish if SNPs could be associated with milk productive traits. The research was conducted on 129 Sarda breed goats from 4 to 5 years old, multiparous, lactating and in their third to fifth lactation. We report nine exonic and seven non-coding regions SNPs within the Sarda goat POU1F1 gene, namely, Ex 1 61 G>C; Ex 1 108 G>A; Ex 3 C>T; Ex 3 92 C>T; Ex 4 110 A>G; Ex 5 34 G>A resulting in Arg213Lys change; IVS4 641 G>A, IVS4 643 A>C, IVS4 659 G>A, IVS4 677 A>C, IVS4 G699Del, IVS4 709 C>G, Ex 6 17 G>A resulting in Arg228Ser change, Ex 6 58 G>T, Ex 6 172 T>C, 3'UTR 110 T>C. A statistically significant association was found between genotype TT, in position 17 of the exon 6 (3.1 % of frequency), and increased milk yield (P < 0.01) while genotype GT (25.6 % of frequency) was associated with a higher fat content. Genotype TT in position 58 of the exon 6 (3.9 % of frequency) was found to be associated with a higher fat (P < 0.01) and protein content (P < 0.05). Twenty-eight haplotypes were detected, but no significant association between the haplotypes and the milk production traits have been found. Our data, as well as providing new SNPs extending the POU1F1 gene characterization, evidence a relationship between polymorphism and milk production traits in Sarda goat breed.
Molecular Biology Reports 11/2012; · 2.93 Impact Factor
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ABSTRACT: The aim of the study was to develop a reliable method for the RNA extraction from milk of Sarda sheep breed and to highlight if the extracted RNA can be used for expression study on mammary genes involved in milk fat synthesis using RT-qPCR. The main result is that a sample of 150 ml of milk provides an optimal amount of RNA (73.5 μg/ml). The highest RNA concentration has been found in the samples analysed within 4 h after collection. The RNA extracted was positively correlated to the number of somatic cells (P < 0.001). The efficiency of the extraction method was confirmed by the results obtained from qPCR which showed a Ct value, for SREBPF1 gene of 26.8 ± 0.15. This research demonstrated that the high-quality of the RNA obtained is suited to use for studies of mammary genes expression in sheep, avoiding any damage caused by mammary gland biopsy.
Molecular Biology Reports 11/2012; · 2.93 Impact Factor
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ABSTRACT: The ovine POU1F1 gene is localized on chromosome 1 and it contains five introns and six exons. In different mammalian species some mutations in different exons are associated with different production traits. The aim of our research was to study the POU1F1 gene nucleotide sequence to detect possible polymorphisms and their relationships with milk productive traits in Sarda breed sheep. The study had been conducted on 140 ewes, 4 or 5 years old coming from a farm located in Sardinia. All the animals were multiparous, lactating and in their third to fifth lactation. Individual milk yield had been recorded monthly and for each sample fat, protein, casein, lactose, and somatic cell count values were analysed. A jugular blood sample was collected from each ewe to perform genomic DNA extraction. PCR, SSCP and sequencing analysis were carried out to examine the six exons to highlight possible SNPs. One-way ANOVA was used to analyse association of variants with milk yield and/or its composition. Two novel SNP were found: 121 C>T in the 5'UTR of the fourth intron fragment and 249 G>A in the 3'UTR of the sixth exon fragment. The statistical analysis did not shown association between milk productive traits and the found polymorphisms. However, further investigations about the promoter region or the prophet genes, like the PROP-1, could clarify its exact role in regulating the productive traits in sheep.
Molecular Biology Reports 02/2012; 39(6):6975-9. · 2.93 Impact Factor
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ABSTRACT: This study evaluated the effect of storage on renneting properties of goat milk investigated using the Formagraph method. Milk samples from 169 goats in three farms (F1, F2 and F3) were analysed during an entire lactation (45, 75, 105, 135 and 165 days in milking DIM), to obtain renneting parameters, both from fresh milk and after storage with Bronopol and freezing at -20°C and -80°C. As regards fresh milk, mean values of clotting time were between 12·51 (45 DIM) and 13·29 min (105 DIM and F2), the curd firming time between 1·77 (45 DIM) and 2·15 min (F1) and curd firmness between 42·09 (165 DIM) and 49·55 mm (45 DIM). No statistical difference was recorded after storage. After regression analysis, all prediction models showed significance value at P<0·001 with the highest R2 value for clotting time, 0·710 (fresh vs. frozen milk at -20°C), and the lowest for clot firmness, 0·281 (fresh vs. frozen milk at -80°C). Results demonstrated that assessment of goat milk coagulation properties using the Formagraph method is also achievable after freezing or Bronopol addition.
Journal of Dairy Research 08/2011; 78(3):343-8. · 1.34 Impact Factor
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ABSTRACT: 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 < 0.01) throughout the lactation. Protein content showed the lowest value (P < 0.01) in June (4.15%). Urea and pH values were fluctuating. FP was lower (P < 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 < 0.01) throughout the lactation and, on the whole, SCC and TMC were not correlated.
Animal Science Journal 10/2010; 81(5):594-9. · 0.86 Impact Factor
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ABSTRACT: The objective of this study was to compare slaughtering performance, carcass characteristics, and meat quality in lambs of two different genotypes. Sixteen crossbred Mouflon x Sarda (M x S) and sixteen pure Sarda breed (S x S) suckling lambs were analysed. Chemical and fatty acid composition were determined on semitendinosus and longissimus dorsi muscles and perirenal and pelvic fats. Dressing percentage was higher (P < 0.01) in M x S group and after histological dissection M x S carcasses had more muscle tissue (P < 0.01) and less separable fat (P < 0.05). Muscles of M x S had less cholesterol than the S x S lambs (P < 0.01). Polyunsaturated/saturated and n −6/ n −3 fatty acids ratios of the intramuscular lipids were optimal in both groups. Indexes of atherogenicity and of thrombogenicity in muscles varied between 0.9 and 1.1. The results indicated several good qualities in both the genotypes, but M x S lambs have some traits which could provide a higher market price.
Meat Science 10/2008; · 2.28 Impact Factor
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ABSTRACT: The reproductive activity of goats bred in temperate latitude follows a seasonal pattern, influenced by annual variation in day length. Daily variation in pineal melatonin secretion is the neuroendocrine signal recognized by animals through the link between this hormone and melatonin receptor 1a (MTNR1A). A total of 345 goats of different breeds (225 Sarda, 30 Saanen, 30 Chamois Coloured, 30 Maltese and 30 Nubian) with a kidding period in October–December or January–March were analysed to verify if a link exists between the structure of the receptor gene and reproductive activity. The main part of exon II of MTNR1A gene was amplified by PCR and then digested with MnlI and RsaI to prove the presence of restriction sites. Sequencing of 20 cloned samples and 20 purified samples permitted comparison with previously published sequences. No polymorphism was found using MnlI enzyme, as all 345 samples showed the cleavage site in position 605 and all the goats were MM genotype. However, using RsaI enzyme, some Sarda goats, showed a polymorphic site in position 53. Nine Sarda goats were R/r genotype, lacking this cleavage site only in one allele, while the other animals, both Sarda and the other breeds, presented the cleavage site in both the alleles and were thus R/R genotype. No r/r genotype was found in any of the breeds. In Sarda goats the allelic frequency was 0.98 for R allele and 0.02 for r allele; genotypic frequency was 96.00% for R/R genotype and 4.00% for R/r genotype. A strong link emerged from statistical analysis ( P < 0.001) between R/r genotype and reproductive activity, which was strongly influenced by photoperiod. Sequencing indicated six nucleotide changes that did not induce any amino acid change. Data showed that polymorphism was present and that it influences reproductive activity only in the Sarda breed.
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ABSTRACT: The goals of the present research were to characterize
the MT1 gene in mouflon, identify possible polymorphisms,
and facilitate a better understanding of their relationship with reproductive activity.
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ABSTRACT: The objective was to obtain a description of genetic diversity at the four casein genes in the Arbi goat, the native goat breed from Tunisia. DNA from 111 Arbi goats (71 females and 40 bucks) was analysed using PCR-based methods. The most frequent allele at the CSN1S1 locus was B, followed by A, F, N, E and 01. The most frequent allele at CSN2 locus was C, followed by A and 01. The CSN2 E allele was not found. The A, C and F alleles prevailed at the CSN1S2 locus, the B allele occurred only in one female goat, in heterozygosis. No subject was found carrying the intermediate CSN1S2 D and the null 0 alleles. The CSN3 A* variant (A + B) prevailed in both sexes on CSN3 C. Five of the estimated haplotypes at the casein gene cluster CSN1S1–CSN2–CSN1S2–CSN3 showed frequency higher than 0.05: BAAA (0.155), ACAA (0.150), BACA (0.114), NCAA (0.086) and FCFA (0.060). Three haplotypes were estimated containing the CSN1S1 E allele, having frequencies lower than 3%. The most frequent haplotypes contained strong alleles at each casein gene, indicating their possible positive effect on milk composition and cheesemaking properties. Unexpectedly, two of the most frequent haplotype combinations contained a null (N) or a weak (F) allele at CSN1S1 locus. Indeed, a casein haplotype combination including the CSN1S1 N allele has not been evidenced so far in goats. Milk produced from animals carrying these haplotypes could be suitable for the production of hypoallergenic drinking milk, owing to the low casein concentration. This information should be taken into account in breeding programs aiming for genetic improvement of milk characteristics.
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ABSTRACT: The aim of this research was to investigate the genetic structure at BMPR 1B, BMP15 and GDF9 prolificacy genes in five sheep breeds reared in Tunisia: Barbarine, Queue Fine de L’Ouest, Noire de Thibar, Sicilo-Sarde and D’man. Genomic DNA of 204 sheep was investigated for the FecB<sup>B</sup> (BMPR 1B), FecX<sup>R</sup>, FecX<sup>H</sup>, FecX<sup>I</sup>, FecX<sup>L</sup>, FecX<sup>G</sup>, FecX<sup>B</sup> (BMP15) and FecG<sup>H</sup> (GDF9) mutations. The sequence variability of the different DNA fragments utilised for genotyping was further investigated by Single Stranded Conformation Polymorphism (SSCP) and sequencing. All the above-mentioned mutations were absent in the five sheep breeds examined. SSCP analysis and sequencing allowed the detection of two nucleotide variations. A non-functional mutation (T/C transition at nt 747 of BMP15 cDNA known as B3) was found at the BMP15 gene, in the Noire de Thibar breed; this mutation was first detected in the Belclare sheep. A new nucleotide change G/A at nt 1159 of BMP15 cDNA, causing the amino acid change A119T in the mature peptide, was detected in the Barbarine breed for the first time. The highly prolific D’man ewes were monomorphic for the absence of all the known prolificacy alleles.
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ABSTRACT: Goat αs<sub>1</sub>-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 αs<sub>1</sub>-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 αs<sub>1</sub>-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.
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ABSTRACT: The aim of this research was to investigate how growth hormone (GH) cortisol and some haematochemical parameters could be modified by the stress caused by the stages of shearing in Sarda breed sheep. Five groups of 10 sheep each were formed. Group A, only separated from the flock; Group B, only tied; Group C, both tied and shorn (animals in these three groups were ewe lambs shorn for the first time); Group D, adult females both tied and shorn; and Group E, adult entire males both tied and shorn (animals in these two groups had been shorn previously). Five blood samples were taken from each animal: the day before treatment (first sample); at the start of the treatment (second sample); in the middle of shearing for Groups C, D and E, 10 min after separation in Group A and 10 min after tying in Group B (third sample); at the end of treatment (fourth sample); and on the day after treatment (fifth sample). Plasma GH levels showed a decrease ( P < 0.01) in Groups A, B, C and D during treatment (third and fourth samples), while Group E only at the end of shearing (fourth sample). In the third sample, the highest GH levels were recorded for Group E ( P < 0.05), while it was recorded in the fourth sample for Groups A and E ( P < 0.05). Cortisol levels showed a clear increase ( P < 0.01) in all groups during treatment, but Group A showed a decrease in the fourth sample in comparison to the third sample. Males in the second, third and fourth sample and Group A only in the fourth sample showed lower cortisol levels when compared with the other groups ( P < 0.05). Plasma glucose levels showed an increase ( P < 0.01) in all groups during treatment but Groups B, C and E showed the highest values ( P < 0.05). Magnesium (Mg) showed an increase in all groups in the third and fourth sample, while sodium (Na), in the same samples, only in Groups B, C and D. Potassium (K) values showed a significant decrease ( P < 0.05) only in Groups C and D at the end of shearing. These results show that GH secretion is influenced by all the stress procedure: separation, tying and shearing. Shearing, even if necessary for animals, causes a significant change of the blood parameters involved in the stress response.
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ABSTRACT: The research was carried out in order to
evaluate α<sub>s1</sub>-casein allele frequency and the possible correlations between its genotype and milk quanti-qualitative
productions in Sarda goat. 600 Sarda goats were utilized, belonging from 24 flocks located in three distinct areas of
Sardinia: Central (Zone 1), South-East Coast (Zone 2) and South-West area (Zone 3). The individual milk samples were
collected during the middle stage of lactation and analysed for fat, protein and lactose percentage, pH value, cryoscopic
index, SCC and TMC. The phenotype analysis of the αs1-casein was carried out by ultra thin layer polyacrylamide gel
isoelectric focusing (UTLIEF) and immunoblotting with specific polyclonal antibodies against α<sub>s1</sub>-casein. The statistical
analysis pointed out significant differences between milk yield, milk quality and genotype frequencies related to the
breeding zone. The following genotype frequencies were detected: BB (63.3%), AB (20.0%), AA (9.0%), EF (3.2%),
BF (2.2%), FF (0.7%), IF (0.7%), AE (0.3%), EE (0.3%) and EI (0.3%). Statistical analysis, performed by ANOVA,
showed interesting differences between the strong genotypes: the significantly higher (P<0.01) milk yield was among
the subjects with AB genotypes (>1 kg/day). The AB and BB genotypes displayed higher (P<0.01) fat (5.51% and
5.47% vs . 5.17%), higher protein values (4.56% and 4.45% vs . 4.20%) than the AA genotype. The lower lactose content
(P<0.01) was registered in the BB genotype (4.93%). The subjects carrying the strong alleles at the homozygous state
showed higher (P<0.01) dry matter percentages and lower pH values (P<0.05) than the other genotypes. No differences
in milk production or chemical composition arose after the other genotypes comparison. The results of the present
research show that milk from Sarda goat is particularly suitable for cheese making. It is possible to apply selection
strategies within this population, directed to the improvement of productions not neglecting milk quality aspects.
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ABSTRACT: The aim of this study was carried out using the productive method for animal welfare
assessment in intensive beef-calf farming. In order to investigate average daily weight gain
(ADWG) in different space-allowance environments, some Italian Friesian calves were
penned in three groups, called A, B and C, made up of 10, 12 and 15 calves respectively.
Each pen measured 25 square meters. Calves were individually weighed on arrival at the
farm, and once every forty five days thereon. Weight data were examined by analysis of
variance (ANOVA). It evidenced statistical differences (P<0,001) among the groups. A and
B Groups showed ADWG higher than that recorded in the ‘overcrowded’ Group C.
Furthermore, all the calves were graded into 1<sup>st</sup> and 2<sup>nd</sup> commercial categories, during the
last weight recording, to achieve an in vivo quality assessment. Commercial grading
showed more 1<sup>st</sup> quality-calves in A and B Groups (9 and 8 respectively) than in Group C,
where only four animals were graded as 1<sup>st</sup> category (P<0.05). Finally, conditioned
pathology rate was higher in Group C. As a result, it could be significant for production
increase to respect the minimum space-allowance, also considering 91/629/EEC directive.
