The hairless (hr) gene is involved in the congenital hypotrichosis of Valle del Belice sheep

Congenital hypotrichosis in mammalian species consists of partial or complete absence of hair at birth. The hairless gene is often responsible for this disorder in men, mice and rats. Recent experimental data on Valle del Belice sheep reared in Sicily for milk production, support the genetic control of the ovine hypotrichosis as a Mendelian recessive trait. The ovine hairless gene was chosen as the candidate gene involved in this disorder. Blood samples were collected from Valle del Belice sheep with the normal and hypotrichotic phenotypes. Almost the entire hairless gene was successfully amplified using the long PCR technique. Unrelated sheep with differing phenotypes were randomly chosen for sequencing the amplified products. Different mutations related to the hypotrichotic phenotype were found in exon 3. In fact, sequencing revealed an A/T transversion at position 739, a G/A transition at position 823, and a C/T transition at position 1312. From these nucleotide exchanges, three substitutions of the processed mature protein were deduced at the amino acid positions 247 (Thr/Ser), 275 (Ala/Thr), and 438 (Gln/Stop). A PCR-SSCP based test was developed in order to detect the last mutation, which is responsible for the hypotrichotic phenotype

Genetic structure of milk protein polymorphisms and effects on milk production traits in a local dairy cattle

The milk protein genetic structure of the Reggiana dairy local cattle with remarkable milk quality properties was analysed and compared with the structure of Italian Friesian and Italian Brown cattle. A total of 1138 individual milk samples from the three breeds were typed by isoelectrofocusing. Polymorphism was found at the as1-casein (CSN1S1), b-casein (CSN2), K-casein (CSN3), and b-lactoglobulin (LGB) loci. High frequencies were observed for some casein alleles in Reggiana (CSN1S1*C, 25%; CSN2*B, (28%) with considerable differences to those observed in a survey carried out 30 years before. A great variation among breeds was observed for casein haplotype frequencies and high linkage disequilibrium was detected at the three polymorphic casein loci CSN1S1-CSN2-CSN3. The prevalent haplotypes were CA2B (23%; Reggiana), BA2A (48%; Italian Friesian) and BA2B (51%; Italian Brown). Higher GST values between breeds were observed when were considered instead of gene frequencies. In the Reggiana cattle, the occurrence of two casein alleles at a rather high frequency (CSN1S1*C, CSN2*B) allowed to estimate the effects of rare casein haplotypes on milk traits. The CA2B haplotype was associated with a significantly higher milk protein and fat content and a significantly higher protein per cent was found also for the BA1B haplotype

Effect of somatic cell count and lactation stage on sheep milk quality

In order to evaluate the effects of mammary health status and lactation phase on the qualitative parameters of ovine<br />milk, 213 individual milk samples were repeatedly collected from 40 primiparous Sarda ewes on a monthly basis. Yield,<br />physico-chemical characteristics, casein fractions quantitative distribution, somatic cell count (SCC), cheese making properties<br />and plasmin-plasminogen activity were determined on each sample. Repeated individual milk SCC were used as a<br />marker of udder health status, allowing the definition of three classes: &ldquo;Healthy&rdquo; (H), &ldquo;Infected&rdquo; (I) or &ldquo;Doubtful&rdquo; (D).<br />Samples were grouped into 4 classes of days in milk (DIM). To evaluate the influence of mammary health status and<br />phase of lactation, a mixed model was performed using the ewe as random effect. Milk physico-chemical parameters were<br />influenced both by the udder health status and by lactation phase. In particular, the udder health status adversely affected<br />&alpha;s1 and &beta;1-casein fractions (P<0.01) and curd firmness (P<0.05). Samples reactive to rennet were 92.73%, 70.65%<br />and 64.60% in &ldquo;H&rdquo;, &ldquo;D&rdquo; and &ldquo;I,&rdquo; respectively. Lactation phase influenced the overall milk composition and technological<br />characteristics. Plasmin activity was higher in the &ldquo;I&rdquo; group than in the others (16.1 vs 11.8 and 11.2 U/ml; P<0.01) and<br />it significantly (P<0.01) increased during lactation. In conclusion, both mastitis and mammary involution are proven to<br />exert a detrimental effect on milk quality since they enhance its endogenous proteolytic activity

The hairless (hr) gene is involved in the congenital hypotrichosis of Valle del Belice sheep

Congenital hypotrichosis in mammalian species consists of partial or complete absence of hair at birth. The hairless gene is often responsible for this disorder in men, mice and rats. Recent experimental data on Valle del Belice sheep reared in Sicily for milk production, support the genetic control of the ovine hypotrichosis as a Mendelian recessive trait. The ovine hairless gene was chosen as the candidate gene involved in this disorder. Blood samples were collected from Valle del Belice sheep with the normal and hypotrichotic phenotypes. Almost the entire hairless gene was successfully amplified using the long PCR technique. Unrelated sheep with differing phenotypes were randomly chosen for sequencing the amplified products. Different mutations related to the hypotrichotic phenotype were found in exon 3. In fact, sequencing revealed an A/T transversion at position 739, a G/A transition at position 823, and a C/T transition at position 1312. From these nucleotide exchanges, three substitutions of the processed mature protein were deduced at the amino acid positions 247 (Thr/Ser), 275 (Ala/Thr), and 438 (Gln/Stop). A PCR-SSCP based test was developed in order to detect the last mutation, which is responsible for the hypotrichotic phenotype

<i>Lactobacillus paracasei</i> CBA L74 Metabolic Products and Fermented Milk for Infant Formula Have Anti-Inflammatory Activity on Dendritic Cells <i>In Vitro</i> and Protective Effects against Colitis and an Enteric Pathogen <i>In Vivo</i>

<div><p>The rapid expansion of commercially available fermented food products raises important safety issues particularly when infant food is concerned. In many cases, the activity of the microorganisms used for fermentation as well as what will be the immunological outcome of fermented food intake is not known. In this manuscript we used complex <i>in vitro</i>, <i>ex-vivo</i> and <i>in vivo</i> systems to study the immunomodulatory properties of probiotic-fermented products (culture supernatant and fermented milk without live bacteria to be used in infant formula).</p><p>We found <i>in vitro</i> and <i>ex-vivo</i> that fermented products of <i>Lactobacillus paracasei</i> CBA L74 act via the inhibition of proinflammatory cytokine release leaving anti-inflammatory cytokines either unaffected or even increased in response to <i>Salmonella</i> typhimurium. These activities are not dependent on the inactivated bacteria but to metabolic products released during the fermentation process. We also show that our <i>in vitro</i> systems are predictive of an <i>in vivo</i> efficacy by the fermented products. Indeed CBA L74 fermented products (both culture medium and fermented milk) could protect against colitis and against an enteric pathogen infection (<i>Salmonella</i> typhimurium). Hence we found that fermented products can act via the inhibition of immune cell inflammation and can protect the host from pathobionts and enteric pathogens. These results open new perspectives in infant nutrition and suggest that <i>L. paracasei</i> CBA L74 fermented formula can provide immune benefits to formula-fed infants, without carrying live bacteria that may be potentially dangerous to an immature infant immune system.</p></div

Fermented milk preparation has anti-inflammatory properties on MoDCs.

<p>MoDCs were infected with <i>Salmonella</i> FB62 (MOI 10:1) for 1 hour in medium without antibiotics or in different infant formulas: non-fermented milk (NFM) and fermented milk (FM) obtained through fermentation of Lactobacillus CBA L74 alone (CBA L74), fermentation in the presence <i>S. thermophilus</i> TH3 (S. th) or double fermentation with CBA L74 and <i>S. thermophilus</i> TH3 (CBA L74+S. th). After washing and 24 h incubations in medium with antibiotics cytokine abundance was evaluated. Panels show % of response of IL-12p70 and IL-10 relative to MoDCs infected with SL1334 in medium without antibiotics. *** p<0,001.</p

<i>Lactobacillus paracaseii</i> CBA L74 supernatant has anti-inflammatory properties on MoDCs.

<p>MoDCs were infected with either <i>Salmonella</i> SL1344 (MOI 10∶1) or <i>Lactobacillus</i> CBA L74 (MOI 10∶1) for 1 hour in medium without antibiotics or in the presence of 2% LB, MRS, FB62 supernatant (Sn FB62) or CBA L74 supernatant in MRS (Sn CBA L74). After washing and 24 h incubations in medium with antibiotics cytokine abundance was evaluated. A. Concentrations of IL-12p70 and IL-10 as determined by ELISA and CBA assays respectively. B. The % of response relative to FB62 infected MoDCs is shown after grouping six experiments together. C. Anti-inflammatory index calculated as ratio of IL-10/IL12p70 concentrations. * p<0,05; *** p<0,001.</p

CBA L74 fermented milk reduces inflammatory mediators expression.

<p>At the end of the experiment described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087615#pone-0087615-g005" target="_blank">Figure 5</a> RNA was extracted from colons and expression levels of Tbet, Gata3, Inf-γ, Il-17a, Il-6, Il-22, Il-33, Il-1β, Kc, Ccl2, Cox2, iNos, Ahr, Indo, Zo-1, Reg3β, Reg3γ were assessed by RT-qPCR. Expression levels are normalized to the housekeeping mRNA Ralp32. Dots represent individual mice measurements, lines represent the average value.</p