Meat quality traits and the expression of tenderness-related genes in the loins of young goats at different ages.

Goat meat is considered healthy because of its low fat content, but it is often rather tough. Tenderness is the most important attribute of quality during meat consumption and there is scarce information about the expression of genes involved in the meat tenderization process in goats. The aim of this trial was to assess certain meat quality traits and the expression, at the messenger RNA (mRNA) and protein levels, of specific genes involved in the tenderization process of the longissimus lumborum (LL) in young male goats (Capra hircus) at different ages. Samples of LL were collected at slaughter from 32 Alpine goats that were divided into three categories: 9 suckling kids (Sk) at 5.4±0.15 weeks of age, 16 chevons (Ch) at 17.1±0.55 weeks of age and 7 post-puberal goats (Pu) at 34.3±2.5 weeks of age. Animal and carcass variables (live weight gain, live weight, carcass weight and fat deposits) and quality traits of meat (lipid content, ultimate pH, color parameters, cooking loss and shear force) were determined. The mRNA abundances of calpain-1 (Capn1), calpain-2 (Capn2), calpastatin (Cast), caspase 3 (Casp3), caspase 9 (Casp9), αB-crystallin (Cryab), heat shock protein 27 (Hsp27), heat shock protein 40 (Hsp40) and heat shock protein 70 (Hsp70) were detected by quantitative PCR. Capn1, Cast, Cryab and Hsp27 protein expression was investigated by ELISA. The Sk group had the leanest carcasses. The meat of the Pu group was the darkest (P<0.05) and the toughest (P<0.05). The redness of meat increased with the age of the goats. The Sk group showed lower mRNA abundances for the Capn2/Cast ratio, Casp3, Cryab, Hsp27, Hsp40 and Hsp70 than the Pu group (P<0.05). Intermediate values were found for the Ch group. Similar results were highlighted for the protein expression of Cryab and Hsp27. The experiment acknowledged a differentiation of the experimental groups based on performance, carcass and meat characteristics, and the genes considered. Moreover, Sk and Pu groups, characterized by a different tenderness of their meat, were clearly discriminated by a different expression of the Hsp.

A survey of selected animal-based measures of dairy cattle welfare in the Eastern Alps: Toward context-based thresholds.

In the Alps, traditional dairy farms are small-scale operations where vertical transhumance from valley indoor housing systems to highland pasture-based systems is still practiced in summer. Vertical transhumance implies a substantial change of environment, available resources, and management practices from one season to another. In such systems, animal-based welfare measures need to be monitored throughout the year to capture the variation of welfare outcomes, based on which targeted welfare management plans can be implemented. Because the Welfare Quality assessment approach has been tailored to indoor housing and intensive farming systems, the European Food Safety Authority recently developed a welfare assessment protocol for small-scale dairy cattle farms adapted after the Welfare Quality framework. The aim of this study was to assess nonbehavioral animal-based measures as defined by this protocol at different time points for transhumant systems in the Alps. In total, 18 animal-based measures were assessed before, during, and after the mountain pasture period in a sample of 67 small-scale dairy cattle farms practicing vertical transhumance in neighboring provinces of Austria, Italy, and Slovenia. Significant differences between assessments were identified for dirtiness of legs and teats, hairless patches, lesions and swellings, claw condition, ocular discharge, and diarrhea whereas BCS, lameness/severe lameness, vulvar discharge, nasal discharge, and hampered respiration were unchanged between seasons. In addition, a benchmarking exercise was carried out to identify relative boundaries (worst quartile thresholds) for each animal-based measure and to contribute to the discussion about achievable welfare outcomes for the 2 husbandry conditions that characterize a transhumant system. Worst quartile thresholds indicated a high prevalence of dirtiness (>80%) when cows were kept indoors, high prevalence of hairless patches (65%) before pasture turnout, and high prevalence of very lean cows (>13%) throughout the assessments. On the other hand, the best quartile thresholds for most clinical conditions suggested that high welfare standards (zero prevalence) are widely achievable in mountain farms practicing vertical transhumance during all assessments. The thresholds identified through benchmarking should serve as the basis for an effective context-based welfare management strategy promoting continuous welfare improvement on-farm.

Effects of grazing cow diet on volatile compounds as well as physicochemical and sensory characteristics of 12-month-ripened Montasio cheese.

The aim of this study was to evaluate the effects of pasture type and cow feeding supplementation level on a 12-mo-ripened Montasio protected designation of origin (PDO) cheese, which is one of the most important PDO cheeses produced in northeast Italy. Cheeses were characterized for volatile compounds, color, mechanical variables, and sensory descriptors. Pasture type significantly affected most of the instrumental variables considered and, as a consequence, sensory properties were affected as well. Cheeses from the pasture characterized by a nutrient-rich vegetation type were higher in protein and lower in fat content. Furthermore, such cheeses, evaluated by a sensory panel, were more intense in color with a more pungent and less cow-like odor, in agreement with what found through instrumental analyses. Supplementation level resulted in less pronounced effects, limited to volatile compounds and texture properties, which were not detected by sensory analysis. The characterization of the 12-mo ripened Montasio cheese reported here is an important step for the valorization of this PDO product.

Volatile compounds and sensory properties of Montasio cheese made from the milk of Simmental cows grazing on alpine pastures.

The aim of this study was to analyze the volatile compounds, physicochemical characteristics, and sensory properties of Montasio, a semicooked pressed cheese, produced from the milk of the dual-purpose Italian Simmental cows grazing on alpine pastures. A total of 72 cows grazing on 2 pastures, which differed in botanical composition (nutrient-rich pasture vs. nutrient-poor pasture), received 2 different levels of supplementation (3.0 vs 1.5 kg/head per day). The experimental cheeses were produced from whole, raw milk and ripened for 60 d. Sixty-one volatile compounds, including alcohols (11), aldehydes (6), ketones (10), lactones (2), esters (6), hydrocarbons (3), carboxylic acids (6), phenolic compounds (4), monoterpenes (7), sesquiterpenes (1), sulfur compounds (4), and amines (1), were detected. The main families in terms of relative weight appeared to be carboxylic acids, esters, and alcohols. A panel of trained assessors described the experimental cheeses as having an intense color; small and evenly distributed eyes; an intense odor and flavor of milk-sour, milk, and cow; and a tender and creamy texture. The pasture type affected the volatile fraction, particularly ketones, phenolic compounds, and terpenes, which are overall higher in nutrient-poor pastures. A slight effect on the sensory analyses, in particular the effect of the cow attribute on odor and flavor, was perceived by the panelists. The cheeses produced on nutrient-rich pasture had higher b* (yellowness) index. These results were consistent with the color evaluation of the sensory panel. In addition, the pasture affected some textural attributes (adhesivity, creaminess, and granules) as perceived by the panelists. Concentrate supplementation, which is required to meet the feeding requirements of grazing cows, had no clear effect on either the volatile compounds or the sensory properties of the cheeses. Thus, at least within levels of integration adopted, it is expected not to alter the organoleptic characteristics of this product.

Effect of linseed addition on the expression of some lipid metabolism genes in the adipose tissue of young Italian Simmental and Holstein bulls.

The objective of this trial was to determine the effect of breed and long-term dietary linseed addition on composition of fatty acids and expression of some genes involved in the lipid metabolism within subcutaneous (s.c.) adipose tissue of young bulls. Italian Simmental and Holstein bulls (n=16/breed) were fed a corn silage-grass hay diet with or without 8% (DM basis) whole ground linseed. Inclusion of linseed, rich in α-linoleic acid (C18:3n-3), increased (P<0.05) the proportions of linolelaidic (C18:2n-6trans), γ-linolenic (C18:3n-6), C18:3n-3, and rumenic (cis9,trans11 conjugated linoleic acid) acids, as well as total n-3 fatty acid, total PUFA, and PUFA:SFA, but decreased (P<0.05) weight percentages of myristic (C14:0), pentadecanoic (C15:0), palmitic (C16:0), palmitelaidic (C16:1n-9trans), and margaric (C17:0) acids, along with n-6:n-3, in the s.c. fat of young bulls. Even though PUFA were similar (P≥0.23) between bull breeds, s.c. fat from Holstein bulls had greater (P<0.05) proportions of tridecylic (C13:0), myristoleic (C14:1) and palmitoleic (C16:1n-9cis) acids and a lower (P<0.05) proportion of margaric (C17:0) acid than s.c. fat from Simmental bulls. Feeding linseed decreased (P<0.05) the expression of stearoyl CoA desaturase (SCD) and the lipoprotein lipase (LPL) gene without affecting (P≥0.19) fatty acid synthase (FASN), leptin (LEP), and PPARγ2 mRNA in the s.c. fat of bulls; however, there was no effect of bull breed (P≥0.11) or interactive effect of breed and linseed (P≥0.23) on gene expression. Expression of PPARγ2 was positively correlated with SCD (r=0.454; P=0.01), LEP (r=0.500; P<0.01), and LPL (r=0.531; P<0.01) mRNA, indicating that PPARγ2 increases the expression of genes involved in lipogenesis.

Effect of whole linseed addition on meat production and quality of Italian Simmental and Holstein young bulls.

The effect of long term dietary linseed addition on performance, carcass characteristics and meat quality of Italian Simmental (IS) and Italian Holstein (HI) young bulls was investigated. Thirty-two animals were assigned to 4 groups following a factorial design: 2 breeds - IS and IH - ×2 diets - containing whole ground linseed (5-8% of DM) and control. IS had greater in vivo performance and carcass characteristics than IH. IS muscle had lower C14:0, C16:0, SFA, higher C18:2n-6 cis, PUFAn-6, PUFA and PUFA/SFA proportion than IH in phospholipids (PL) fraction. Linseed inclusion did not affect animal's performance and carcass characteristics. In muscle PL, linseed increased C20:0, C22:0, C23:0, C20:5n-3 and decreased C20:4n-6, PUFAn-6/n-3, PUFAn-6 concentration. Linseed decreased C14:0, C16:0 proportion in neutral lipids (NL) and increased total PUFAn-3, C18:3n-3 proportions both in NL and PL fraction. However, these differences were relatively low from a quantitative point of view.

Association analyses of single nucleotide polymorphisms in the LEP and SCD1 genes on the fatty acid profile of muscle fat in Simmental bulls.

The aim was to investigate the effect of the genetic polymorphisms of leptin (LEP) and stearoyl-CoA desaturase (SCD1) genes on the fatty acid (FA) composition of the muscle of 103 Simmental bulls. Ten single nucleotide polymorphisms (SNP) were detected in exons 2 and 3 of the LEP gene, two of them encoding non-synonymous mutations. Allelic substitution effects of all the SNP on 28 single fatty acids, monounsaturated (MUFA) and polyunsaturated (PUFA) and desaturation indexes were estimated. Both the SCD1 SNP, as well as three SNP of the leptin gene, affected, to different extents, the desaturation of FA into MUFA. Because it was previously proposed that leptin's metabolic action involves down-regulation of SCD1, it is possible that, beyond the mere additive effect of SCD1 gene on FA desaturation, the non-synonymous mutations in the leptin gene also contribute to the variability of FA composition in muscle fat.

Fatty acid composition and sensory properties of Italian Simmental beef as affected by gene frequency of Montbéliarde origin.

The aim of this study was to investigate the effect of Montbéliarde (Mb) gene frequency on fatty acid composition and sensory properties of Italian Simmental (IS) steaks (longissimus thoracis m.). Twenty-seven bulls belonging to three strains with different percentages of Mb genes: traditional (ISt), without Mb ascendants (ISt=0% Mb genes), cross-strain (ISmt=37.5-50% Mb genes), Montbéliarde strain (ISm=87.5-100% Mb genes) and balanced for stearoyl Co-A desaturase genotype were considered. ISt has the highest C20:4 n-6 (P<0.01), C22:4 n-6 (P<0.05) and total PUFA n-3 level (P<0.01), while ISt and ISmt have higher C18:3 n-3 (P<0.05) and slightly lower MUFA (P=0.08) than ISm. Sensory tests indicated that the three experimental groups can be differentiated; moreover, ISmt meat is perceived as less hard (P<0.01), less chewable (P<0.01) and less fibrous (P<0.05) than ISt meat.

Characterization of SURF-1 expression and Surf-1p function in normal and disease conditions.

Loss-of-function mutations of the SURF-1 gene have been associated with Leigh syndrome with cytochrome c oxidase (COX) deficiency. Mature Surf-1 protein (Surf-1p) is a 30 kDa hydrophobic polypeptide whose function is still unknown. Using antibodies against a recombinant, hemagglutinin-tagged Surf-1p, we have demonstrated that this protein is imported into mitochondria as a larger precursor, which is then processed into the mature product by cleaving off an N-terminal leader polypeptide of approximately 40 amino acids. By using western blot analysis with specific antibodies, we showed that Surf-1p is localized in and tightly bound to the mitochondrial inner membrane. The same analysis revealed that no protein is present in cell lines harboring loss-of-function mutations of SURF-1, regardless of their type and position. Northern blot analysis showed the virtual absence of specific SURF-1 transcripts in different mutant cell lines. This result suggests that several mutations of SURF-1 are associated with severe mRNA instability. To understand better whether and which domains of the protein are essential for function, we generated several constructs with truncated or partially deleted SURF-1 cDNAs. None of these constructs, expressed into Surf-1p null mutant cells, were able to rescue the COX phenotype, suggesting that different regions of the protein are all essential for function. Finally, experiments based on blue native two-dimensional gel electrophoresis indicated that assembly of COX in Surf-1p null mutants is blocked at an early step, most likely before the incorporation of subunit II in the nascent intermediates composed of subunit I alone or subunit I plus subunit IV. However, detection of residual amounts of fully assembled complex suggests a certain degree of redundancy of this system.

Genomic structure, promoter characterisation and mutational analysis of the S100A7 gene: exclusion of a candidate for familial psoriasis susceptibility.

We have recently assigned a locus for familial psoriasis (PS) susceptibility to the region containing the epidermal differentiation complex gene cluster on chromosome 1q21. Gene S10OA7 maps within this cluster and is reported to be markedly over-expressed in the skin lesions of psoriatic patients. In order to analyse S100A7 as a candidate for PS susceptibility, we have determined its genomic structure regarding exon-intron boundaries and the transcription start site. The gene is organised in three exons and two introns, spanning 2.7 kb. The 5' flanking region contains AP1- and Sp1-binding motifs and a TATA box. We have performed functional assays by using the beta-galactosidase gene as a reporter and have confirmed that this region has strong promoter activity. To search for nucleotide variation within S100A7, we have designed a set of primers to amplify each exon and the gene promoter. Polymerase chain reaction products from 15 unrelated PS patients selected from 1q-linked pedigrees and 25 normal controls have been characterised by single-strand conformation polymorphism and direct sequencing techniques. These analyses have revealed the presence of two polymorphisms in the promoter region (-559G/A and -563 A/G), neither of which shows preferential association with the disease. Our results indicate that S100A7 can be excluded as a candidate for PS susceptibility.