Quantitative PCR reveals the frequency and distribution of 3 indigenous yeast species across a range of specialty cheeses.

Indigenous microorganisms are important components of the complex ecosystem of many dairy foods including cheeses, and they are potential contributors to the development of a specific cheese's sensory properties. Among these indigenous microorganisms are the yeasts Cyberlindnera jadinii, Pichia kudriavzevii, and Kazachstania servazzii, which were previously detected using traditional microbiological methods in both raw milk and some artisanal specialty cheeses produced in the province of Québec, Canada. However, their levels across different cheese varieties are unknown. A highly specific and sensitive real-time quantitative PCR assay was developed to quantitate these yeast species in a variety of specialty cheeses (bloomy-rind, washed-rind, and natural-rind cheeses from raw, thermized, and pasteurized milks). The specificity of the quantitative PCR assay was validated, and it showed no cross-amplification with 11 other fungal microorganisms usually found in bloomy-rind and washed-rind cheeses. Cyberlindnera jadinii and P. kudriavzevii were found in the majority of the cheeses analyzed (25 of 29 and 24 of 29 cheeses, respectively) in concentrations up to 104 to 108 gene copies/g in the cheese cores, which are considered oxygen-poor environments, and 101 to 104 gene copies/cm2 in the rind. However, their high abundance was not observed in the same samples. Whereas C. jadinii was present and dominant in all core and rind samples, P. kudriavzevii was mostly present in cheese cores. In contrast, K. servazzii was present in the rinds of only 2 cheeses, in concentrations ranging from 101 to 103 gene copies/cm2, and in 1 cheese core at 105 gene copies/g. Thus, in the ecosystems of specialty cheeses, indigenous yeasts are highly frequent but variable, with certain species selectively present in specific varieties. These results shed light on some indigenous yeasts that establish during the ripening of specialty cheeses.

Tissue-specific profiling reveals modulation of cellular and mitochondrial oxidative stress in normal- and low-birthweight piglets throughout the peri-weaning period.

Weaning is known to induce important nutritional and energetic stress in piglets. Low-birthweight (LBW) piglets, now frequently observed in swine production, are more likely to be affected. The weaning period is also associated with dysfunctional immune responses, uncontrolled inflammation and oxidative stress conditions that are recognized risk factors for infections and diseases. Mounting evidence indicates that mitochondria, the main cellular sources of energy in the form of adenosine 5' triphosphate (ATP) and primary sites of reactive oxygen species production, are related to immunity, inflammation and bacterial pathogenesis. However, no information is currently available regarding the link between mitochondrial energy production and oxidative stress in weaned piglets. The objective of this study was to characterize markers of cellular and mitochondrial energy metabolism and oxidative status in both normal-birthweight (NBW) and LBW piglets throughout the peri-weaning period. To conduct the study, 30 multiparous sows were inseminated and litters were standardized to 12 piglets. All the piglets were weighted at day 1 and 120 piglets were selected and assigned to 1 of 2 experimental groups: NBW (n = 60, mean weight of 1.73 ± 0.01 kg) and LBW piglets weighing less than 1.2 kg (n = 60, 1.01 ± 0.01 kg). Then, 10 piglets from each group were selected at 14, 21 (weaning), 23, 25, 29 and 35 days of age to collect plasma and organ (liver, intestine and kidney) samples. Analysis revealed that ATP concentrations were lower in liver of piglets after weaning than during lactation (P < 0.05) thus suggesting a significant impact of weaning stress on mitochondrial energy production. Oxidative damage to DNA (8-hydroxy-2'-deoxyguanosine, 8-OHdG) and proteins (carbonyls) measured in plasma increased after weaning and this coincides with a rise in enzymatic antioxidant activity of glutathione peroxidase (GPx) and superoxide dismutase (SOD) (P < 0.05). Mitochondrial activities of both GPx and SOD are also significantly higher (P < 0.05) in kidney of piglets after weaning. Additionally, oxidative damage to macromolecules is more important in LBW piglets as measured concentrations of 8-OHdG and protein carbonyls are significantly higher (P < 0.05) in plasma and liver samples, respectively, than for NBW piglets. These results provide novel information about the nature, intensity and duration of weaning stress by revealing that weaning induces mitochondrial dysfunction and cellular oxidative stress conditions which last for at least 2 weeks and more severely impact smaller piglets.

Folic acid supplementation reduces multigenerational sperm miRNA perturbation induced by in utero environmental contaminant exposure.

Persistent organic pollutants (POPs) can induce epigenetic changes in the paternal germline. Here, we report that folic acid (FA) supplementation mitigates sperm miRNA profiles transgenerationally following in utero paternal exposure to POPs in a rat model. Pregnant founder dams were exposed to an environmentally relevant POPs mixture (or corn oil) ± FA supplementation and subsequent F1-F4 male descendants were not exposed to POPs and were fed the FA control diet. Sperm miRNA profiles of intergenerational (F1, F2) and transgenerational (F3, F4) lineages were investigated using miRNA deep sequencing. Across the F1-F4 generations, sperm miRNA profiles were less perturbed with POPs+FA compared to sperm from descendants of dams treated with POPs alone. POPs exposure consistently led to alteration of three sperm miRNAs across two generations, and similarly one sperm miRNA due to POPs+FA; which was in common with one POPs intergenerationally altered sperm miRNA. The sperm miRNAs that were affected by POPs alone are known to target genes involved in mammary gland and embryonic organ development in F1, sex differentiation and reproductive system development in F2 and cognition and brain development in F3. When the POPs treatment was combined with FA supplementation, however, these same miRNA-targeted gene pathways were perturbed to a lesser extend and only in F1 sperm. These findings suggest that FA partially mitigates the effect of POPs on paternally derived miRNA in a intergenerational manner.

Growth performance of piglets during the first two weeks of lactation affects the development of the intestinal microbiota.

The aim of this study was to evaluate the effect of newborn piglet weight gain during the first 2 weeks of lactation on the luminal and mucosal microbiota of the ileum and colon. The microbiota from high-weight-gain (HWG) and low-weight-gain (LWG) 2-week-old piglets was characterized by amplicon length heterogeneity PCR (LH-PCR) and compared using diversity indices and multivariate statistical analyses. At birth, LWG piglets weighted in average 0.26 kg less than HWG piglets (p = .002). The weight difference between LWG and HWG piglets increased with time and reached 2.1 kg after 16 days of lactation (p < .0001). Based on these growth performance differences, estimated colostrum and milk intake was greater in HWG than in LWG piglets (p < .0001). Analysis of the LH-PCR data of the microbiota using non-metric multidimensional scaling (NMS) and blocked multiresponse permutation procedure (MRBP) revealed that the microbiota of the HWG and LWG piglets tended to differ in ileal mucosa (p = .097) and differed in colonic lumen (p = .024). The microbiota of HWG piglets had higher levels of Bacteroidetes, Bacteroides and Ruminoccocaceae, and lower proportions of Actinobacillus porcinus and Lactobacillus amylovorus when compared with those of LWG piglets. As the weight gain of nursing piglets is highly correlated with the amount of ingested colostrum and milk, the results strongly suggest that colostrum and milk intake in the first 2 weeks of life influenced the development of the gut microbiota.

Use of the spectrophotometric color method for the determination of the age of skin lesions on the pig carcass and its relationship with gene expression and histological and histochemical parameters.

The presence of lesions on the pig carcass is an indicator of poor animal welfare and has economic impact as it downgrades the carcass value. The assessment of the age of lesions on the carcass may help identify risk factors and ultimately prevent their occurrence. The aim of this study was to assess the age of lesions on pig carcasses through spectrophotometric color evaluation and to relate the results with gene expression and histological and histochemical parameters. A total of 96 barrows were mixed 4 times over 3 d before slaughter and 80 lesions were selected after skin lesion observations to define 4 age categories: < 7 h (T1), 7-25 h (T2), 25-30 h (T3), and 49-54 h (T4). A nonlesioned skin area was used as a control. At slaughter, 3 biopsies per lesion and control skin were taken immediately after bleeding for analyses of gene expression (, , , , , , , , , ), skin histological characteristics (inflammation, erosion or ulceration, and necrosis), and enzyme activity (alkaline phosphatase and adenosine triphosphatase). The number of lesions was counted on each carcass, and the color was assessed visually by a pictorial chart and instrumentally through a spectrophotometer. Delta values (Δ) were calculated as the difference between the value of the lesion and the value of the control for all measures, except for the histological analysis. Results indicated that visual color observation was not sufficiently accurate to discriminate lesions by time of infliction ( > 0.10), while the spectrophotometer ΔL* and Δa* values variation allowed the identification of < 7 h or > 25 h old lesions ( < 0.05). Similarly, the expression of , , , , and genes was higher ( < 0.05) in < 7 h old lesions, while gene expression was higher ( < 0.05) in < 25 h old lesions. As for the histological analysis, the severity of inflammation was correlated with the age of the lesion (lower score in < 7 h old lesions and higher score in > 25 h old lesions; < 0.05). To conclude, the spectrophotometric color assessment of the carcass lesions at slaughter appears to be a reliable method to discriminate between fresh and older lesions on the carcass at the abattoir.

Oxytocin injections in the postpartal period affect mammary tight junctions in sows.

The potential impacts of injecting oxytocin (OXY) to sows in the early postpartum period on the quality of mammary tight junctions, milk composition, and immune status of sows and piglets were studied. Postparturient sows received i.m. injections of either saline (control [CTL]; = 10) or 75 IU of OXY ( = 10). Injections were given twice daily (0800 and 1630 h) starting on d 2 of lactation (i.e., between 12 and 20 h after birth of the last piglet), totaling 4 injections. Milk samples were obtained before the first injection (d 2 morning [AM]), before the second injection (d 2 afternoon [PM]), and on d 4 PM and d 5 PM. Blood samples were obtained from sows before milking on d 2 AM, d 2 PM, and d 5 PM. On d 5 of lactation, a blood sample was obtained from 3 piglets per litter. Circulating concentrations of prolactin, IGF-I, lactose, and IgA in sows did not differ between treatments at any time ( > 0.10), but OXY sows had less IgG than CTL sows ( < 0.01) on d 2 PM before the second OXY injection. There were differences in milk composition on d 2 PM, with OXY sows having more IGF-I ( < 0.01), solids ( < 0.05), protein ( < 0.01), energy ( < 0.05), and IgA ( < 0.01) and a greater Na:K ratio ( < 0.01) than CTL sows. These differences were not seen in the next 2 milk samples, except for protein and IgA that still tended ( < 0.10) to be greater in OXY vs. CTL sows on d 4 PM (for protein) and on d 5 PM (for IgA) after the last injection. Milk lactose content was lower in OXY vs. CTL sows on d 5 PM ( < 0.01). Values for immunoglobulin immunocrit, IgG, IgA, and IGF-I in piglet blood did not differ between treatments ( > 0.10). Injecting OXY to sows in the early postpartum period increased leakiness of the mammary tight junctions, improved composition of early milk, and may potentially affect immune status of neonatal piglets.

African-American patients with cancer Talking About Clinical Trials (TACT) with oncologists during consultations: evaluating the efficacy of tailored health messages in a randomised controlled trial-the TACT study protocol.

INTRODUCTION: Low rates of accrual of African-American (AA) patients with cancer to therapeutic clinical trials (CTs) represent a serious and modifiable racial disparity in healthcare that impedes the development of promising cancer therapies. Suboptimal physician-patient consultation communication is a barrier to the accrual of patients with cancer of any race, but communication difficulties are compounded with AA patients. Providing tailored health messages (THM) to AA patients and their physician about CTs has the potential to improve communication, lower barriers to accrual and ameliorate health disparities. OBJECTIVE: (1) Demonstrate the efficacy of THM to increase patient activation as measured by direct observation. (2) Demonstrate the efficacy of THM to improve patient outcomes associated with barriers to AA participation. (3) Explore associations among preconsultation levels of: (A) trust in medical researchers, (B) knowledge and attitudes towards CTs, (C) patient-family member congruence in decision-making, and (D) involvement/information preferences, and group assignment. METHODS AND ANALYSIS: First, using established methods, we will develop THM materials. Second, the efficacy of the intervention is determined in a 2 by 2 factorial randomised controlled trial to test the effectiveness of (1) providing 357 AA patients with cancer with THM with 2 different 'depths' of tailoring and (2) either providing feedback to oncologists about the patients' trial THM or not. The primary analysis compares patient engaged communication in 4 groups preconsultation and postconsultation. ETHICS AND DISSEMINATION: This study was approved by the Virginia Commonwealth University Institutional Review Board. To facilitate use of the THM intervention in diverse settings, we will convene 'user groups' at 3 major US cancer centres. To facilitate dissemination, we will post all materials and the implementation guide in publicly available locations. TRIAL REGISTRATION NUMBER: NCT02356549.

Effect of a post-weaning diet supplemented with functional feed additives on ileal transcriptome activity and serum cytokines in piglets challenged with lipopolysaccharide.

This study evaluated the potential of a weanling diet supplemented with trace minerals, vitamins, prebiotics, essential oils, antioxidants and bovine colostrum (BC) to modulate the inflammatory response of low-weight (LW) and high-weight (HW) piglets challenged with lipopolysaccharide (LPS). At weaning (20±1 d), litters from 32 sows were assigned to four groups: control diet (CTL), CTL plus dietary supplements (DS) or the antibiotic chlortetracycline (ATB), or DS plus BC in place of plasma proteins in the weanling diet (DS+BC). At 37 d (T0), two LW and two HW piglets were bled to evaluate ex vivo cytokine production by LPS activated peripheral blood mononuclear cells (PBMCs). In parallel, LW and HW piglets received intraperitoneal LPS and were bled at slaughter at 4h (T4) or 18h (T18) post-injection. Ileal tissues from these piglets and two unchallenged medium weight (MW) piglets per treatment were excised and analyzed by microarray. At T0, cytokine production of LPS-activated PBMCs was not affected by dietary treatments. At T4 after LPS challenge, serum concentrations of TNF-α, IL-6, IL-8, and IL-10 were increased in all piglets (P<0.01). Interestingly, the LW piglets had a higher TNF-α level than the HW piglets did (P=0.05). Dietary treatments had no effect on the piglet serum concentration of these cytokines neither at T4 nor at T18. Microarray data and QPCR analysis reveal that several genes were differentially expressed in the LPS-challenged piglets in comparison with the two control MW piglets (P<0.001). However, the dietary treatments had a slight effect on the ileal gene expression of the T4 and T18 LPS-challenged piglets when all piglets were included in the analysis. But when body weight (LW and HW) was considered as a fixed effect, the microarray analysis showed that the expression of 54 genes was differentially modulated by the dietary treatments in the T4 and T18 LPS-challenged LW piglets (P<0.05) while in HW piglets no difference was observed. QPCR analyses confirm that the level expression of several genes was reduced in LW piglets fed DS or DS+BC diet compared with ATB piglets. In conclusion, LPS challenge induced a transitional inflammation in weanling piglets that was characterized by increased blood-circulating cytokines and gut transcriptome activity. Results also suggest that the weanling diet supplemented with feed additives attenuated the ileal gene response to the LPS challenge, an effect that was more pronounced in the LW piglets.

Including sheath effects in the interpretation of planar retarding potential analyzer's low-energy ion data.

The interpretation of planar retarding potential analyzers (RPA) during ionospheric sounding rocket missions requires modeling the thick 3D plasma sheath. This paper overviews the theory of RPAs with an emphasis placed on the impact of the sheath on current-voltage (I-V) curves. It then describes the Petite Ion Probe (PIP) which has been designed to function in this difficult regime. The data analysis procedure for this instrument is discussed in detail. Data analysis begins by modeling the sheath with the Spacecraft Plasma Interaction System (SPIS), a particle-in-cell code. Test particles are traced through the sheath and detector to determine the detector's response. A training set is constructed from these simulated curves for a support vector regression analysis which relates the properties of the I-V curve to the properties of the plasma. The first in situ use of the PIPs occurred during the MICA sounding rocket mission which launched from Poker Flat, Alaska in February of 2012. These data are presented as a case study, providing valuable cross-instrument comparisons. A heritage top-hat thermal ion electrostatic analyzer, called the HT, and a multi-needle Langmuir probe have been used to validate both the PIPs and the data analysis method. Compared to the HT, the PIP ion temperature measurements agree with a root-mean-square error of 0.023 eV. These two instruments agree on the parallel-to-B plasma flow velocity with a root-mean-square error of 130 m/s. The PIP with its field of view aligned perpendicular-to-B provided a density measurement with an 11% error compared to the multi-needle Langmuir Probe. Higher error in the other PIP's density measurement is likely due to simplifications in the SPIS model geometry.

Effects of probiotics Pediococcus acidilactici strain MA18/5M and Saccharomyces cerevisiae subsp. boulardii strain SB-CNCM I-1079 on fecal and intestinal microbiota of nursing and weanling piglets.

In this study, the influence of (PA) and subsp. (SCB) on fecal and intestinal microbiota of piglets during lactation and after weaning was monitored. Forty sows and their litters were used and allocated to the following dietary treatments: 1) PA, 2) SCB, 3) a mixture of the 2 probiotics (PA+SCB), 4) antibiotics (ATB), and 5) control (CTRL). Four weeks before parturition, probiotic-treated sows started receiving a daily probiotic dose of at least 2.5 × 10 cfu mixed in 500 g of feed until the end of lactation. The other groups were fed a diet without probiotics and ATB. Two days after birth, piglets received, daily, 1 × 10 cfu of the same probiotics as their mother. At weaning (d 21), these piglets were fed a basal diet enriched with the same probiotics whereas piglets from untreated litters were fed the basal diet with or without ATB. Two piglets per litter were randomly chosen to evaluate the influence of treatments on fecal microbial composition (d 10 and 28) and on ileum and colon microbiota at d 37. The microbiota was characterized by culture on selective media and by 16S rRNA gene diversity assessment using the terminal RFLP technique and clone library analysis to evaluate diversity index and phylum affiliation. Terminal RFLP profiles were also analyzed to determine differences in microbial composition between animals receiving different treatments and to identify diet-specific terminal restriction fragments (TRF) using pairwise multiresponse permutation procedures (MRPP) and indicator species analysis. Before weaning, administration of probiotics to sows and piglets had minor effect on fecal microbiota of piglets. Most modulatory effects of probiotics on ileum and colon microbiota were observed on d 37. Results revealed that PA or ATB treatments reduced ileal microbiota diversity compared with the CTRL ( < 0.05) and promoted the establishment of Firmicutes whereas SCB consumption positively influenced the establishment of the Porphyromonadaceae and Ruminococcaceae bacterial families in the colon. Moreover, pairwise MRPP analysis indicated that ileum bacterial communities of pigs treated with PA or ATB differed from those of CTRL pigs ( < 0.05). In conclusion, PA and SCB supplements, respectively, influenced, in a strain-dependent manner, the ileum and colon microbiota of weaned piglets. Results also suggest that PA and SCB have the potential as feed additives to modulate bacterial populations associated with gut health.

Colostrum whey down-regulates the expression of early and late inflammatory response genes induced by Escherichia coli and Salmonella enterica Typhimurium components in intestinal epithelial cells.

Pathogenic invasion by Escherichia coli and Salmonellae remains a constant threat to the integrity of the intestinal epithelium and can rapidly induce inflammatory responses. At birth, colostrum consumption exerts numerous beneficial effects on the properties of intestinal epithelial cells and protects the gastrointestinal tract of newborns from pathogenic invasion. The present study aimed to investigate the effect of colostrum on the early and late inflammatory responses induced by pathogens. The short-term (2 h) and long-term (24 h) effects of exposure to heat-killed (HK) E. coli and Salmonella enterica Typhimurium on gene expression in the porcine intestinal epithelial cell (IPEC-J2) model were first evaluated by microarray and quantitative PCR analyses. Luciferase assays were performed using a NF-κB-luc reporter construct to investigate the effect of colostrum whey treatment on the activation of NF-κB induced by HK bacteria. Luciferase assays were also performed using NF-κB-luc, IL-8-luc and IL-6-luc reporter constructs in human colon adenocarcinoma Caco-2/15 cells exposed to dose-response stimulations with HK bacteria and colostrum whey. Bovine colostrum whey treatment decreased the expression of early and late inflammatory genes induced by HK bacteria in IPEC-J2, as well as the transcriptional activation of NF-κB-luc induced by HK bacteria. Unlike that with colostrum whey, treatment with other milk fractions failed to decrease the activation of NF-κB-luc induced by HK bacteria. Lastly, the reduction of the HK bacteria-induced activation of NF-κB-luc, IL-8-luc and IL-6-luc by colostrum whey was dose dependent. The results of the present study indicate that bovine colostrum may protect and preserve the integrity of the intestinal mucosal barrier in the host by controlling the expression levels of early and late inflammatory genes following invasion by enteric pathogens.

Homocysteine metabolism, growth performance, and immune responses in suckling and weanling piglets.

Homocysteine (Hcy), an intermediary sulfur AA, is recognized as a powerful prooxidant with deleterious effects on physiological and immune functions. In piglets, there is an acute 10-fold increase of plasma concentrations of homocysteine (pHcy) during the first 2 wk of life. This project aimed to maximize pHcy variations within physiological ranges using typical supplies of folates and vitamin B12 (B12) to sows and piglets. Growth, immune response, and Hcy metabolism of piglets were studied until piglets reached 56 d of age. Third-parity sows were randomly assigned to a 2 × 2 split-plot design with 2 dietary treatments during gestation and lactation, S(-) (1 mg/kg folates and 20 µg/kg B12, n = 15) and S(+) (10-fold S(-) levels, n = 16), and 2 treatments to piglets within each half litter, intramuscular injections (150 µg) of B12 (P(+)) at d 1 and 21 (weaning) and saline (P(-)). Within each litter of 12 piglets, 3 P(+) and 3 P(-) piglets were studied for growth and Hcy metabolism, and the others were studied for immune responses. During lactation, plasma B12 decreased and was transiently greater in S(+) vs. S(-) piglets on d 1 and P(+) vs. P(-) piglets on d 7 (sow treatment × age and piglet treatment × age; P < 0.05). From 14 to 21 d of age, pHcy was 33% lower in S(+)P(+) vs. S(-)P(-) piglets (sow treatment × piglet treatment interaction; P < 0.05). At 56 d of age, hepatic B12 was greater and pHcy was lower for P(+) vs. P(-) piglets (P < 0.05). No treatment effect was observed on growth except for a lower postweaning G:F in S(+)P(-) piglets than in others (sow treatment × piglet treatment interaction; P < 0.05). Positive correlations were observed between pHcy and growth (r > 0.29, P < 0.05) before and after weaning. Antibody responses to ovalbumin and serum tumor necrosis factor-α were not affected by treatments, but postweaning serum IL-8 peaked earlier in S(-)P(-) vs. S(+)P(+) piglets (piglet treatment × age; sow treatment × piglet treatment interaction, P < 0.05). Proliferation of lymphocytes in response to the mitogen concanavalin A tended to be lower in culture media supplemented with sera from S(-) vs. S(+) piglets (P = 0.081) and P(-) vs. P(+) piglets (P = 0.098), and the reduction of response was more marked (P < 0.05) with high (>21 µM) compared to medium (17 to 21 µM) or low (<17 µM) pHcy. In conclusion, the present vitamin supplements to sows and/or piglets produced variations of pHcy that were not apparently harmful for growth performance of piglets. The greater pHcy, particularly prevalent in S(-) and/or P(-) piglets, had negative effects on some indicators of immune responses, suggesting that these young animals may be immunologically more fragile.

Increased blood-circulating interferon-γ, interleukin-17, and osteopontin levels in bovine paratuberculosis.

Paratuberculosis-infected cattle initially develop an effective cell-mediated immune response that declines as the disease progresses. Blood is one of best sources for characterizing the inflammatory status of infected cows and for studying mediators related to chronic diseases. The aim of this study was to evaluate the cow-level association between blood cytokine concentration, the influence of serum on immune cell proliferation, and dairy cows naturally infected with Mycobacterium avium ssp. paratuberculosis (MAP). Positive animals (n=41) from 19 herds were selected on the basis of 2 positive fecal culture results and divided into 2 groups: single-positive, or serum ELISA-negative cows (n=32), and double-positive, or cows that gave positive results for both mycobacterial culture and serum ELISA (n=9). Negative animals (n=39) were selected from paratuberculosis-negative herds in which at least 80% of the animals had been diagnosed as negative by fecal culture and ELISA and that did not produce positive results during the 2-yr study. Analysis of plasma levels of the cytokines IL-4, IL-10, IL-17, IFN-γ, and osteopontin was performed, revealing distinct patterns. The ELISA-positive cows with MAP shedding had similar plasma concentrations of IL-4 and IL-10 but elevated levels of IFN-γ, IL-17, and osteopontin, which is indicative of inflammatory disease in these subclinical positive cows. In vitro MAP infection of bovine macrophages showed increased gene expression of tumor necrosis factor-α, IL-1ß, IL-6, IL-23, and transforming growth factor-ß as early as 6h postinfection for all of the cytokines involved in the establishment of a T-helper type-17 immune response. To determine the systemic influence of serum on immune cell functions, lymphoproliferation assays were also performed in presence of JD serum. The serum from shedding cows showed 15% less proliferation. These results indicate that infected cows have a lower systemic capacity to maintain a protective immune response and that, as the disease progresses, an emerging T-helper type-17 immune response is established.

The effect of birth weight and age at tail docking and ear notching on the behavioral and physiological responses of piglets.

Selection for high prolificacy has resulted in litters comprising a large number of low-birth-weight (LBW) piglets. Given their presence in over 75% of litters and increased mortality rate, it is clear that a greater understanding of LBW piglet management is required for both animal welfare and productivity. In this study, we compared the effects of tail docking and ear notching LBW and average-birth-weight (ABW) piglets at 1 or 3 d of age on suckling, behavior, passive transfer of immunoglobulins, and growth. Six piglets per litter from 20 litters (n = 120 piglets) were used in a 2 × 2 complete block design. Piglets were weighed at birth and designated as LBW (0.6 to 1.0 kg) or ABW (≥ 1.2 kg) and "processed" (tail docked and ear notched) at either 1 or 3 d of age. Vocalizations were recorded during the procedures. The acute behavioral responses were observed for 10 min after the procedure. Piglets were observed for 6 h after birth and after the procedure to determine their presence at nursing bouts. On d 5, blood samples were collected to determine concentrations of serum immunoglobulins (IgA and IgG) and IGF-I. Piglet weights were recorded at birth and on d 5, 14, and 21. During the procedures, LBW piglets produced fewer (P = 0.03) calls than ABW piglets. Piglets from either birth weight category produced a similar number (calls/s; P = 0.29) of high-frequency calls (≥ 1,000 Hz), which are indicative of pain and distress, although the average frequency (Hz) of these calls was greatest (P = 0.05) for ABW piglets processed on d 3. Immediately following the procedures, LBW piglets spent more (P = 0.005) time dog-sitting and less (P = 0.005) time lying than ABW piglets. When observed with the sow, LBW males spent more (P = 0.001) time alone and had the lowest (P = 0.007) attendance at nursing bouts compared with LBW females and all ABW piglets. Concentrations of serum IgA (P = 0.06) and IgG (P = 0.04) and plasma IGF-I (P = 0.003) were lower for LBW than ABW piglets regardless of age of processing although the magnitude of these differences was likely not of biological significance. Average-birth-weight piglets may be less reactive to the acute effects of the procedures on d 1 than on d 3. Given the decreased likelihood of a LBW piglet surviving to weaning (P = 0.001), delaying processing until 3 d of age for LBW piglets may eliminate unnecessary procedures.

A gene expression programme induced by bovine colostrum whey promotes growth and wound-healing processes in intestinal epithelial cells.

Bovine colostrum is well known for its beneficial properties on health and development. It contains a wide variety of bioactive ingredients that are known to promote a number of cellular processes. Therefore the use of colostrum whey as a feed additive to promote intestinal health has been proposed, yet little is known about mechanisms implicated in its beneficial properties on intestinal epithelial cells. In the present paper, casein were removed from bovine colostrum and the remaining liquid, rich in bioactive compounds, was evaluated for its capacity to modulate cellular processes in porcine intestinal epithelial cell line IPEC-J2 and human colon adenocarcinoma cell line Caco-2/15. First, we verified the effect of colostrum whey and cheese whey on processes involved in intestinal wound healing, including cell proliferation, attachment, morphology and migration. Our results showed that colostrum whey promoted proliferation and migration, and decreased specifically the attachment of Caco-2/15 cells on the culture dish. On the other hand, cheese whey induced proliferation and morphological changes in IPEC-J2 cells, but failed to induce migration. The gene expression profile of IPEC-J2 cells following colostrum whey treatment was evaluated by microarray analysis. Results revealed that the expression of a significant number of genes involved in cell migration, adhesion and proliferation was indeed affected in colostrum whey-treated cells. In conclusion, colostrum specific bioactive content could be beneficial for intestinal epithelial cell homoeostasis by controlling biological processes implicated in wound healing through a precise gene expression programme.

Impact of a long photoperiod during lactation on immune status of piglets.

The effects of a long photoperiod treatment around parturition and throughout lactation on immune status of piglets were studied. Sows were assigned to 2 light regimens: i) standard short photoperiod (SP, n = 17), 8 h of daily light from d 112 of gestation until d 23 of lactation; and ii) long photoperiod (LP, n = 17), 23 h of daily light from d 112 of gestation to d 4 of lactation and 16 h thereafter. In front of the crates, under the side heat lamps and behind the sow, light intensities were 59 ± 5, 109 ± 6, and 44 ± 6 lx, respectively. On d 15 of lactation and at weaning (d 23), 2 piglets of similar BW per litter were selected and immunized intramuscularly with ovalbumin (OVA). Blood samples (5 mL serum and 10 mL whole blood) were taken at d 15 and d 23 of lactation, and at d 30, 37, and 44 of age after weaning to evaluate the antibody response to OVA and measure phagocytosis, lymphocyte proliferative response, and different circulating blood lymphocyte populations of piglets. Results showed that phagocytosis was increased in piglets submitted to LP (P < 0.05). A treatment × time interaction (P < 0.001) indicated that SP piglets developed a better IgG response to OVA than LP piglets. The percentage of B lymphocytes was also increased (P = 0.02) in SP piglets compared with piglets exposed to LP during lactation; the lymphocyte response to OVA tended to be enhanced (P = 0.07) over time in SP piglets. Different subpopulations of CD8+ lymphocytes were markedly increased in SP piglets at 23 d of age compared with piglets exposed to LP (treatment × time: P < 0.05). These results suggest that exposure of piglets to LP during lactation seems to reduce the capacity of piglets to develop a strong immune response to novel antigens. This may have important consequences on the ability of piglets to resist an infection after weaning.

Quantitative measurement of FMRP in blood platelets as a new screening test for fragile X syndrome.

The fragile X syndrome usually results from CGG repeats expansion and methylation of the FMR1 gene leading to the absence of expression of its encoded protein, fragile X mental retardation protein (FMRP). Therefore, its diagnosis is traditionally based on the detection of these molecular alterations. As an alternative, FMRP-based screening methods have been proposed over the years. Most of them are based on immunohistochemistry analyses applied to a restricted number of lymphocytes (100) or hair roots (10-20) with limited diagnosis potential. In this study, we describe a truly quantitative approach using a new model, the blood platelet, which can be recovered easily with very high purity (99.9%). FMRP levels in platelets were first measured in a control population (n = 124) and reference values were established. FMRP measurements were also performed in confirmed fragile X subjects. Receiver operating characteristic curve analysis has shown that our test can easily discriminate fragile X males and females from controls (area under curve, AUC = 0.948). Cognitive functions were also assessed in these individuals using age-specific Wechsler Intelligence Scales for Children and the Vineland Adaptive Behavior Scales. A proportional relationship between FMRP levels, intelligence quotient and adaptive behavior was observed among fragile X individuals, suggesting that our test would be able to detect fragile X cases and may predict cognitive functions.

Exploring suppression subtractive hybridization (SSH) for discriminating Lactococcus lactis ssp. cremoris SK11 and ATCC 19257 in mixed culture based on the expression of strain-specific genes.

AIM: An approach based on quantitative reverse transcriptase PCR (RT-qPCR) was developed for monitoring two strains of lactococci in co-culture in milk by measuring the expression of specific genes identified by suppression subtractive hybridization (SSH). METHODS AND RESULTS: SSH was used to identify strain-specific genes of Lactococcus lactis ssp. cremoris SK11 and ATCC 19257. RT-qPCR was then employed to validate gene specificity and compare the expression of selected specific genes (glycosyltransferase and amidase genes for L. lactis ssp. cremoris ATCC 19257 and a hypothetical protein for SK11) identified by SSH. The time profile of changes in gene expression relative to ldh transcription differed between pure and mixed cultures as well as between media. At the stationary phase, gene expression of mixed cultures in GM17 attained the highest proportion of ldh transcription while mixed cultures in milk peaked at the postexponential phase. Strain ratios expressed as RNA proportion appear to favour SK11 in GM17 medium, while ATCC 19257 dominated in milk co-cultures. CONCLUSIONS: This approach was useful to determine the contribution of strain SK11 in relation to strain ATCC 19257 during co-culture in milk compared to rich medium. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability to track the metabolic contribution of each lactococcal strain during fermentation of milk or cheese ripening will extend our understanding of the impact of process parameters on the production performance of strains.

Dietary supplementation with different forms of flax in late gestation and lactation: Effects on sow and litter performances, endocrinology, and immune response.

The impact of feeding flax as seed, meal, or oil to late-pregnant and lactating sows on hormone concentrations, immune responses, and sow and litter performances was studied. Sixty sows were fed 1 of 4 diets from 68 d of gestation until 21 d of lactation. Diets were standard (CTL; n = 15); 10% flaxseed supplementation (FS; n = 16); 6.5% flaxseed meal supplementation (FSM; n = 14); and 3.5% flaxseed oil supplementation (FSO, n = 15). On d 88 and 101 of gestation, sows were immunized against ovalbumin (OVA). Jugular blood samples were obtained on d 62, 88, and 110 of gestation and on d 2 and 21 of lactation to measure concentrations of estradiol, prolactin, and progesterone as well as antibody (Ab) against OVA (anti-OVA), lymphocyte proliferation, and lymphocyte production of interferon-gamma (IFN-gamma). Milk samples were obtained on d 3 and 20 of lactation. One piglet per litter was slaughtered on d 1 for compositional analyses, and a jugular blood sample was obtained for anti-OVA analyses. Remaining piglets were weighed on d 2, 7, 14, 21 (weaning), 28, and 56. Circulating hormone concentrations in sows were not affected by treatment overall (P > 0.1). On d 20 of lactation, milk from FS, FSM, and FSO sows had more protein than that from CTL sows (P < 0.01). The FSM piglets weighed more on d 56 (P < 0.05) than FS and FSO piglets. Carcasses of 1-d-old FSM piglets also had greater glycogen (P < 0.001) and DM (P = 0.05) contents than FS and FSO piglets, but organ weights and circulating concentrations of glucose and IGF-I did not differ (P > 0.1). In CTL sows, IFN-gamma production decreased between d 101 of gestation and d 2 of lactation, whereas, in FS sows, IFN-gamma production increased (P < 0.01). Anti-OVA for the whole experimental period was greater in FS than in FSO sows (P < 0.05). Concentrations of anti-OVA in milk on d 3 of lactation and lymphocyte proliferative responses were not affected by treatments (P > 0.1). Serum concentrations of anti-OVA in 2-d-old piglets that gained BW during the first 24 h after birth were greater (P < 0.01) in FS, FSM, and FSO litters than in CTL litters and percent mortality on d 2 and 21 postpartum was less (P < 0.05) for FS, FSM, and FSO litters compared with CTL. Therefore, feeding flax to sows may have beneficial effects on immune resistance of piglets and feeding FSM improved postweaning growth of piglets.