Changes occurring in immune responsiveness of single- and twin-bearing Comisana ewes during the transition period.

Changes induced by twin and single lambing in the immune response of 16 periparturient Comisana ewes were studied. Cell-mediated immune responses were evaluated by means of skin tests performed from 3 wk before and up to d 35 after parturition. At d 21 and 7 before lambing, the sheep received an intramuscular injection of the antigen keyhole limpet hemocyanin (KLH), to which the animals had not been previously exposed, to determine their humoral immune response. Starting 3 wk before lambing and up to d 35 postlambing, the ewes were sampled to determine the plasma concentrations of anti-KLH antibody (IgG), IL-6, and IL-1 beta. From parturition through d 35 postpartum, individual milk samples were collected for determination of anti-KLH IgG titers and IL-6 and IL-1beta concentrations by means of a capture ELISA. The number of lambs born affected IL-6 concentrations in ewe plasma; IL-6 secretion always was higher in ewes birthing twins than in single-lambing ewes. Apart from the number of lambs born, the concentrations of plasma IL-6 in ewes were higher at lambing than at d 21 antepartum and at d 35 postpartum. An interaction of number of lambs born x time of sampling was observed for plasma antibody titers to KLH. The IgG concentrations were significantly higher in single-bearing ewes than in twin-bearing ewes before parturition and were very similar across groups after parturition. A time effect was found for the cell-mediated immune response and for anti-KLH IgG concentrations in milk, such that at parturition, cellular responses were lowest, and the anti-KLH IgG concentration was highest. A significant correlation was found for IgG titers to KLH in plasma and milk. Results indicate that IL-6 concentrations in blood can be considered a reliable indicator of stress connected to lambing and that the mammary gland is a microenvironment unrelated to blood stream with respect to interleukins expression. In contrast, a relationship was found for the IgG secretions in milk and blood, which suggests that the assessment of humoral immune status may be combined with milking routine in dairy animals

Airspace effects on the yield and quality of ewe milk.

Abstract Three groups of 12 midlactating Comisana ewes were housed in separate rooms of the same building and assigned to treatments of low (LV, 4.1m 3 ), medium (MV, 5.6m 3 ), or high (HV, 7.3m 3 ) airspace/animal. The concentrations of airborne microorganisms in the experimental rooms were measured twice weekly at 0930 and 1630. Ewe milk yield was recorded daily. Individual milk samples were analyzed weekly for milk composition, coagulating properties, somatic cell concentration (SCC), and polymorphonuclear neutrophil leukocyte count (PMNLC), and fortnightly for bacteriological characteristics; samples with more than 10 6 somatic cells/ml were cultured for mastitis-related pathogens. The LV and MV treatments resulted in higher relative humidity and air concentrations of staphylococci than the HV treatment. Greater amounts of air mesophilic bacteria were also found in the LV than in the HV room. Ewes in the HV group gave greater yields of milk than those in the LV and MV groups. LV milk also had a lower casein content than HV milk. Significant interactions of treatment x time were found for milk protein and fat content as well as for clotting time and clot firmness, with LV milk having the poorest composition and deteriorated renneting ability during the last 3 wk of the trial. The HV ewes had lower SCC and PMNLC and psychrotroph counts in their milk than LV and MV ewes and smaller amounts of mesophilic bacteria and fecal coliforms than LV animals. Subclinical mastitis occurred in two ewes of the LV and one of the MV groups, while no cases were recorded in the HV group. Results suggest that airspace is a critical factor in dairy sheep housing and indicate that a volume allocation of less than 7m 3 /animal may adversely affect the performance and health of the lactating ewe

Relationship between cortisol response to stress and behavior, immune profile, and production performance of dairy ewes.

The existence of a relationship between cortisol levels, after an acute stress, and behavioral activities, immunological profile, and production performance in sheep was studied. An initial flock of 30 Comisana ewes was involved in the experiment, and each of the 30 ewes was individually subjected to an isolation test in a novel environment. Subsequently, from the initial flock, 2 groups of 8 Comisana ewes were each retrospectively selected, and the animals were divided, according to their cortisol concentration 10 min after the isolation test, into high cortisol (HC) ewes, having a peak of cortisol concentration >90 ng/mL (average: 119.3 ng/mL +/- 11.8), and low cortisol (LC) ewes having a peak of cortisol concentration <80 ng/mL (average: 52.4+/-11.8). During the isolation test, the behavior of each animal was video-recorded and behavioral activities were registered. Blood samples were collected before the isolation test, immediately after the test (10 min), and at 60, 120, 300 min, 24 h, and 48 h after the test to evaluate percentages of T-helper (CD4(+)) and T-cytotoxic (CD8(+)) cells, CD4(+)/CD8(+) ratio, and IL-1beta and IL-6 levels. The ewes were milked for 3 d after the isolation test to determine cortisol levels and IL-1beta and IL-6 concentrations in whey. Milk yield was recorded at each milking, and milk samples were analyzed for pH, nutritional parameters, renneting properties, and somatic cell count. During the isolation test, HC ewes exhibited a shorter duration of movement and fewer bleats than LC ewes. The average plasma IL-1beta concentration was higher in HC than in LC ewes. The average whey IL-1beta and IL-6 concentrations were higher in whey from HC ewes than in LC ewes. A positive correlation emerged between plasma and whey IL-1beta concentrations. The average CD4(+)/CD8(+) ratio in blood was lower in HC than in LC ewes. Time from isolation affected the CD4(+)/CD8(+) ratio: at 120 min, the CD4(+)/CD8(+) ratio increased compared with that at 10 min after isolation and then decreased until 300 min after isolation. On average, ewes with low cortisol concentrations showed higher milk production and lower SCC than ewes with high cortisol concentrations. Results suggest that plasma cortisol concentration is connected to the behavioral response and immune competence of dairy ewes and cytokine concentrations. Both whey IL-1beta and IL-6 can be considered reliable indicators of the magnitude of hypothalamic-pituitary-adrenal axis activation. The stress-induced changes in CD4(+)/CD8(+) ratio are critical for controlling disease incidence and planning appropriate vaccination programs. High reactivity of the hypothalamic-pituitary-adrenal axis is also associated with a reduction in milk production and an increased predisposition to develop intramammary inflammatory processes

Alterations in sheep peripheral blood mononuclear cell proliferation and cytokine release by polyunsaturated fatty acid supplementation in the diet under high ambient temperature.

The aim of this study was to investigate the effects of polyunsaturated fatty acid (PUFA) supplementation from different sources in the diet of dairy sheep under high ambient temperatures on ex vivo lymphocyte proliferation and inflammatory responses. The experiment was carried out during summer: 32 Comisana ewes were divided into 4 groups of 8. The FS group was supplemented with whole flaxseed, the AG group was supplemented with Ascophyllum nodosum, the FS+AG group was supplemented with a combination of flaxseed and A. nodosum. The fourth group (CON group) was a control and received a diet containing no supplement. The average maximum temperature was around 33°C during wk 2 and 3, whereas the mean temperature never decreased below 26°C. Following 15 d of treatment with respective diets, peripheral blood mononuclear cells (PBMC) from sheep who received a diet supplemented with A. nodosum had impaired cell proliferation responses and IL-6 production after mitogen stimulation compared with PBMC from FS+AG sheep. In addition, PBMC from AG sheep displayed impaired cell proliferation compared with cells from the CON group. The FS+AG cells produced lower levels of IL-10 than CON cells, and higher IL-6 than AG and CON cells. Results demonstrated that the supplementation with PUFA from different sources in a sheep's diet can influence their immunological responses under high ambient temperatures depending on the composition of fatty acid supplementation. In particular, synergistic effects of different PUFA from flaxseed and A. nodosum, simultaneously administrated in the sheep diet, were observed on activation of inflammation response

Overground walking training with the i-Walker, a robotic servo-assistive device, enhances balance in patients with subacute stroke: a randomized controlled trial

Background: Patients affected by mild stroke benefit more from physiological overground walking training than walking-like training performed in place using specific devices. The aim of the study was to evaluate the effects of overground robotic walking training performed with the servo-assistive robotic rollator (i-Walker) on walking, balance, gait stability and falls in a community setting in patients with mild subacute stroke. Methods: Forty-four patients were randomly assigned to two different groups that received the same therapy in two daily 40-min sessions 5 days a week for 4 weeks. Twenty sessions of standard therapy were performed by both groups. In the other 20 sessions the subjects enrolled in the i-Walker-Group (iWG) performed with the i-Walker and the Control-Group patients (CG) performed the same amount of conventional walking oriented therapy. Clinical and instrumented gait assessments were made pre- and post-treatment. The follow-up observation consisted of recording the number of fallers in the community setting after 6 months. Results: Treatment effectiveness was higher in the iWG group in terms of balance improvement (Tinetti: 68.4 +/- 27.6 % vs. 48.1 +/- 33.9 %, p = 0.033) and 10-m and 6-min timed walking tests (significant interaction between group and time: F(1,40) = 14.252, p = 0.001; and F(1,40) = 7.883, p = 0.008, respectively). When measured, latero-lateral upper body accelerations were reduced in iWG (F = 4.727, p = 0.036), suggesting increased gait stability, which was supported by a reduced number of falls at home. Conclusions: A robotic servo-assisted i-Walker improved walking performance and balance in patients affected by mild/moderate stroke, leading to increased gait stability and reduced falls in the community

Macro- And micro-nutrient composition and antioxidant activity of Chickpea and Pea Accessions

Epidemiological studies reported an inverse association between the consumption of legumes and the incidence of age-related diseases. This trend could be attributed to the presence of antioxidant compounds, especially phenolic and flavonoid compounds. In this paper, five pea (Pisum sativum L.) and twelve chickpea (Cicer arietinum L.) accessions, having different characteristics and geographical origin, were characterised in terms of antioxidant activity, as well as macro- and micro-nutrient composition. The antioxidant activity has been evaluated using both DPPH (2,2-diphenyl-1-picrylhydra-zyl) and ABTS (2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) radical scavenging capacity assays. Chickpea and pea accessions showed a different behaviour in the presence of these different radicals. Chickpea accessions were characterised by significantly higher DPPH• scavenging activity, while peas showed a significantly higher value of antioxidant activity evaluated using the ABTS assay. Pea accessions had the highest content of total phenolic compounds, Zn, and Cu. A positive correlation was found between some minerals, such as Zn, Cu and P, and the ABTS•+ scavenging activity. Black and brown chickpea accessions showed significantly higher contents of anthocyanins, Mn, Mg, and Ca, which were positively correlated with the antioxidant activity assessed with the DPPH assay. Despite the dataset investigated in our study included a limited number of accessions, it was possible to highlight the influence of the chemical composition on the antioxidant activity due to the high phenotypic diversity found between the accessions, emphasising the importance of selecting the antioxidant activity assay according to the matrix to be evaluated

Free-amino acid metabolic profiling of visceral adipose tissue from obese subjects

Interest in adipose tissue pathophysiology and biochemistry have expanded considerably in the past two decades due to the ever increasing and alarming rates of global obesity and its critical outcome defined as metabolic syndrome (MS). This obesity-linked systemic dysfunction generates high risk factors of developing perilous diseases like type 2 diabetes, cardiovascular disease or cancer. Amino acids could play a crucial role in the pathophysiology of the MS onset. Focus of this study was to fully characterize amino acids metabolome modulations in visceral adipose tissues (VAT) from three adult cohorts: (i) obese patients (BMI 43-48) with metabolic syndrome (PO), (ii) obese subjects metabolically well (O), and (iii) non obese individuals (H). 128 metabolites identified as 20 protein amino acids, 85 related compounds and 13 dipeptides were measured by ultrahigh performance liquid chromatography-tandem mass spectroscopy (UPLC-MS/MS) and gas chromatography-/mass spectrometry GC/MS, in visceral fat samples from a total of 53 patients. Our analysis indicates a probable enhanced BCAA (leucine, isoleucine, valine) degradation in both VAT from O and PO subjects, while levels of their oxidation products are increased. Also PO and O VAT samples were characterized by: elevated levels of kynurenine, a catabolic product of tryptophan and precursor of diabetogenic substances, a significant increase of cysteine sulfinic acid levels, a decrease of 1-methylhistidine, and an up regulating trend of 3-methylhistidine levels. We hope this profiling can aid in novel clinical strategies development against the progression from obesity to metabolic syndrome

Novel transglutaminase 1 mutations in patients affected by lamellar ichthyosis

Lamellar Ichthyosis (LI) is a form of congenital ichthyosis that is caused by mutations in the TGM1 gene that encodes for the transglutaminase 1 (TG1) enzyme. Functional inactivation of TG1 could be due to mutations, deletion or insertions. In this study, we have screened 16 patients affected by LI and found six new mutations: two transition/transversion (R37G, V112A), two nonsense mutations and two putative splice site both leading to a premature stop codon. The mutations are localized in exons 2 (N-terminal domain), 5, 11 (central catalytic domain), and none is located in the two beta-barrel C-terminal domains. In conclusion, this study expands the current knowledge on TGM1 mutation spectrum, increasing the characterization of mutations would provide more accurate prenatal genetic counselling for parents at-risk individuals