Sow environment during gestation: part I. Influence on maternal physiology and lacteal secretions in relation with neonatal survival

In pig husbandry, pregnant females are often exposed to stressful conditions, and their outcomes on maternal and offspring health have not been well evaluated. The present study aimed at testing whether improving the welfare of gestating sows could be associated with a better maternal health during gestation, changes in the composition of lacteal secretions and improvement in piglet survival. Two contrasted group-housing systems for gestating sows were used, that is, a French conventional system on slatted floor (C, 49 sows) and an enriched system using larger pens on deep straw (E, 57 sows). On the 105^th days of gestation (DG105), sows were transferred into identical farrowing crates on slatted floor. Saliva was collected from all sows on DG35, DG105 and DG107. Blood samples were collected on DG105 from all sows and on the 1^st day of lactation (DL1) from a subset of them (C, n=18; E, n=19). Colostrum and milk samples were collected from this subset of sows at farrowing (DL0) and DL4. Saliva concentration of cortisol was greater in C than in E sows at DG35 and DG105, and dropped to concentrations comparable to E sows after transfer into farrowing crates (DG107). On DG105, plasma concentrations of haptoglobin, immunoglobulins G (IgG) and A (IgA), blood lymphocyte counts and plasma antioxidant potential did not differ between groups (P > 0.10), whereas blood granulocyte count, and plasma hydroperoxide concentration were lower in E than in C sows (P < 0.05). Concentrations of IgG and IgA in colostrum and milk did not differ between the two groups. The number of cells did not differ in colostrum but was greater in milk from E than C sows (P < 0.05). Pre-weaning mortality rates were lower in E than C piglets (16.7% v. 25.8%, P < 0.001), and especially between 12 and 72 h postpartum (P < 0.001). Plasma concentration of IgG was similar in E and C piglets on DL4. In conclusion, differences in salivary cortisol, blood granulocyte count and oxidative stress markers between groups suggested improved welfare and reduced immune solicitation during late gestation in sows of the E compared with the C system. However, the better survival observed for neonates in the E environment could not be explained by variations in colostrum composition.     

A multi-herd study shows that saliva is more than a reflection of serum biomarkers in pigs

This study evaluates if biomarkers of porcine health status in saliva samples is a mere reflection of serum to detect disease in pigs under field conditions. Four farms from the same commercial company were included to obtain samples from animals with different pathological conditions. A total of 10 healthy animals and 10–15 animals from each farm with clinical symptoms of the disease were sampled for paired saliva and blood during a veterinary clinical visit. The biomarker panel included acute-phase proteins (APPs), C-reactive protein (CRP), haptoglobin (Hp), an inflammatory marker, adenosine deaminase (ADA), the total antioxidant capacity (TAC), the levels of essential trace elements, copper (Cu) and zinc (Zn), and the measurement of the total protein content (TP). After detailed statistical analysis, the results showed that saliva could replace serum for APP measurements since a good agreement has been observed between the concentrations of APPs in both body fluids. For any other biomarker, no agreement between the concentrations quantified in serum and saliva samples was observed visually. However, salivary ADA and TP concentrations were statistically significantly higher in the diseased, whereas the statistical tests with serum concentrations were inconclusive. Furthermore, greater differentiation between healthy and diseased animals could be observed when the distribution of biomarkers was analysed in saliva than in other serum samples. The diagnostic power to discriminate between healthy and diseased pigs is similar in saliva and in serum samples. Preliminary regression models may offer an optimal combination of biomarkers for disease detection in saliva (Hp, CRP, and TAC) and serum (Hp, CRP, and Cu), which demands less labour, sample, and financial cost for saliva determinations. The contradictory results observed for TAC, Cu, and Zn levels between body fluids indicate a need for further studies. To sum up, saliva-based biomarkers instead of serum-based biomarkers could contribute to more efficient detection of diseased animals.     

Fish oil rich in eicosapentaenoic acid and docosahexaenoic acid in sow diets modifies oxylipins and immune indicators in colostrum and milk

Colostrum and milk are the first nutrient sources for newborn piglets. In addition, n-3 fatty acids (FAs) and their oxygenated derivatives (oxylipins) have the capacity to modulate immune components. The aim of the current study was to include a fish oil rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in sow diets to promote an increase of anti-inflammatory molecules in colostrum and milk to benefit piglets. Thirty-six sows were randomly assigned from insemination to the end of lactation to either a control diet with animal fat (15 g/kg in gestation and 30 g/kg in lactation) or an n-3 diet in which animal fat was totally (gestation) or half (lactation) replaced by an equivalent amount of solid fish oil. Performance of sows and piglets was monitored during the study. Colostrum and milk samples were obtained after the birth of the first piglet and at weaning, respectively. From all samples (n = 18 per treatment), FAs were quantified by gas chromatography and immunoglobulins and cytokines by ELISA. Three samples per treatment were randomly selected to analyse oxylipin composition by liquid chromatography-tandem mass spectrometry. In colostrum and in milk, the n-3 FA (P = 0.020 and P < 0.001), particularly EPA (P < 0.001 and P < 0.001) and DHA (P < 0.001 and P < 0.001), and also their oxygenated derivatives were increased in samples from sows fed n-3 diet. Fish oil had no effect on immunoglobulin concentrations, but reduced tumour necrosis factor α (TNFα) (P = 0.011) and a tendency to reduce interleukin 10 (IL10) (P = 0.059) were observed in milk. In conclusion, fish oil in sow diets increased n-3 FA, particularly EPA and DHA, and their oxygenated derivatives in colostrum and milk, reducing TNFα and IL10 in milk.     

Immunoglobulin A N-glycosylation Presents Important Body Fluid-specific Variations in Lactating Mothers

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Highlights

• •Comparative N-Glycome analysis of IgA from breast milk, saliva, and plasma of lactating participants revealed 89 glycan compositions that may correspond to over 250 structures.
• •Specific N-glycan structures can be used to distinguish IgA from breast milk, saliva, and plasma.
• •Delivery mode influenced the maternal IgA N-glycan profile in saliva and plasma.

     

Piglets’ behaviour and performance in relation to sow characteristics

The importance of maternal care in commercial pig production is largely ignored. The sow has little possibility to interact with her piglets, and piglets are often subjected to early weaning or artificial rearing. This study aimed to investigate aspects of physiological and behavioural maternal provisioning that contribute to offspring outcomes. We hypothesised that better maternal care and nutritional provisioning would relate positively to piglet immunity, growth and behaviour. Nineteen sows and their litters were studied in free-farrowing pens. Oxytocin and tumour necrosis factor-α in colostrum/milk and salivary cortisol were sampled from sows throughout lactation. Sows were assessed for dominance rank, response to handling, maternal defensiveness, suckling initiation and termination, posture and sow-piglet contact. Piglets were weighed, measured for body mass index (BMI) and sampled for blood (Immunoglobulin G; at birth). After weaning, they experienced a human approach test (HAT) and novel object test. Correlations were explored between individual sow characteristics, individual piglet outcomes, and between sow characteristics and piglet outcomes averaged by litter. Significant correlations between sow and piglet factors were analysed at the litter level in mixed models with piglet outcomes as response variables and sow characteristics as predictor variables, while accounting for sow parity, litter size and batch. Litters grew faster when their sow had lower cortisol values (P = 0.03), while sows with lower cortisol levels had more successful suckling bouts and engaged in greater amounts of sow-piglet contact. Litters had a lower BMI at weaning when the sow had a higher milk fat percentage at d3. Litters of the most dominant sows took longer to approach the human in the HAT, while litters of sows with higher cortisol at d0 took longer to approach the novel object when assessed on correlations (r = 0.82, P < 0.001) but not when the model accounted for parity and litter size (P = 0.35). Only some of the measured nutritive and non-nutritive sow factors influenced litter performance and behaviour, with parity and litter size also playing a role. Given the continued increase in litter size, but also the interest in loose-housed lactation pens for sows, further research on sows’ maternal investment and how it can be optimised is warranted.     

Effects of yeast culture supplementation from late gestation to weaning on performance of lactating sows and growth of nursing piglets

Dietary yeast culture supplementation can contribute to the performance and health of sows and piglets, but few studies have focused on the relationships between the effects of yeast culture and gut microbiota. This study investigated the effect of yeast culture (Saccharomyces cerevisiae) supplementation from late gestation to weaning on the reproductive performance of lactating sows and their faecal microbiota. One hundred and six purebred Landrace sows, of parities two to six were selected and randomly assigned to a control (CON) and yeast culture supplementation (YC) groups based on parity and back fat thickness. The YC sows were individually fed with yeast culture at a dose of 24 g/d from day 90 of gestation to parturition and 40 g/d during lactational period. Blood samples were collected from sows on d 110 of gestation and at weaning at day 21 of lactation for plasma hormone and immunoglobulin analysis. Colostrum and milk on day 20 of lactation were collected for composition analysis. Faecal samples from sows on day 110 of gestation and day 20 of lactation were collected for short-chain fatty acid and faecal microbial analysis. Results showed that the farrowing performance of YC sows did not differ significantly from the CON group (P > 0.05). The average daily feed intake by the YC group during the lactation period was significantly increased by 9.98% (P = 0.004), the weaning-to-oestrus interval was shortened by 0.96 d (P = 0.046) and average daily weight gain of piglets increased by 7.14% (P = 0.036) compared with the CON group. Yeast culture supplementation also significantly improved the average daily milk yield in the first week of lactation (P = 0.035), lactose content in colostrum (P = 0.046), protein (P = 0.033) and DM (P < 0.001) content of milk. In the YC group, concentrations of plasma ghrelin (P = 0.02) and IgG (P = 0.015) were increased compared with the CON group, while that of glucagon-like peptide-1 was decreased (P = 0.006) on d 110 of gestation. The 16S rRNA gene sequencing showed that faecal microbiota changed at taxonomic levels with yeast culture addition (P < 0.05). Dietary yeast culture supplementation from late gestation to lactation improved feed intake, immunity status, milk yield, milk quality and faecal microbiota of sows, resulting in the improved growth performance of piglets.     

mRNA expression of genes involved in fatty acid utilization in skeletal muscle and white adipose tissues of sows during lactation

Rodents are able to lower fatty acid utilization in liver and muscle during lactation in order to spare fatty acids for the production of milk triacylglycerols, an effect which is mediated by a down-regulation of peroxisome proliferator-activated receptor α (PPARα). The present study was performed to investigate whether similar fatty acid sparing effects are developing in lactating sows. We considered PPARα and its target genes involved in fatty acid utilization in biopsy samples from muscle and adipose tissue of lactating compared to non-lactating sows. In muscle, PPARα target genes involved in fatty acid utilization were up-regulated during lactation indicating that the fatty acid utilization in muscle was increased. Activation of PPARα was probably due to increased concentrations of non-esterified fatty acids in plasma observed in the lactating sows. In contrast to muscle, PPARα and its target genes involved in β-oxidation in white adipose tissue were down-regulated in early lactation. Overall, the present study shows that sows, unlike rats, are not able to reduce the fatty acid utilization in muscle in order to spare fatty acids for milk production. However, fatty acid oxidation in adipose tissue is lowered during early lactation, an effect that might be helpful to conserve fatty acids released from adipose tissue for the delivery into other tissues, including mammary gland, via the blood.     

The adipose triglyceride lipase,adiponectin and visfatin are downregulated by tumor necrosis factor-α (TNF-α) in vivo

Inflammatory cytokines have been linked to obesity-related insulin resistance. To investigate the effect of TNF-α, an inflammatory cytokine, on insulin action, C57BL/6J mice were treated with TNF-α for 7 days after which we examined the in vivo effects of TNF-α on glucose tolerance and insulin sensitivity with IV glucose tolerance tests and hyperinsulinemic-euglycemic clamps. In addition, we analyzed the in vivo effect of TNF-α on several metabolism-related genes and adipocytokines implicated in the development of insulin resistance. TNF-α treatment resulted in markedly increased fasting blood glucose, insulin and free fatty acids (FFA) levels and reduced glucose tolerance. During the clamps, the rates insulin-stimulated whole body (G_Rd) and skeletal muscle glucose uptake (MGU) and insulin’s ability to suppress hepatic glucose production (HGP) were decreased in TNF-α treated animals, indicating insulin resistance. In addition, both PPARγ and ATGL mRNA expression in adipose tissues as well as ATGL protein levels in plasma were downregulated. Moreover, adipose mRNA expression and plasma protein levels of adiponectin and visfatin were significantly down-regulated. We conclude that the alterations of PPARγ, ATGL, adiponectin and visfatin may contribute to the development of insulin resistance mediated by TNF-α.     

Dimensions of maternal behaviour characteristics in domestic and wildxdomestic crossbred sows

We examined the maternal behaviour of seven domestic and seven wildxdomestic primiparous sows during 10 days post partum to investigate two questions: (1) Did maternal behaviour change during domestication? (2) Can the interindividual variability of maternal behaviour be subsumed into a few dimensions of maternal temperament? We recorded: (a) willingness to leave the nest for food on Day 2; (b) reaction to a playback of squeezed piglet distress vocalisation on Day 2; (c) spontaneous nursing behaviour and spontaneous lying-down behaviour on Day 5 (from an overnight video recording); (d) reactions to playbacks of various piglet distress vocalisations on Day 6 and (e) reactions to a human in the 'nest' with piglets on Day 9. Moreover, data on baseline cortisol saliva concentration and its increase during a brief transportation period and novel environment challenge at the age of 5 months were available. Crossbred sows did not differ from domestic ones in any aspect of maternal behaviour except for a higher tendency to terminate final massage during nursings and a higher frequency of changing posture from lying to standing and back during the night. Factor analysis (based on correlation matrix of 11 behaviour and cortisol variables calculated for all 14 sows after removing the effect of breed) indicated that 82% of the variability in the data could be explained by three factors: first, 'calmness' on which low night time frequency of major posture changes, carefulness of lying-down behaviour and high propensity to remain in nursing position after milk ejection loaded positively while cortisol concentrations during challenge loaded negatively; second, 'protectiveness' with high loadings of the reaction scores to the playbacks of piglet distress calls and the human presence near the piglets; and third, 'nursing activity' which was strongly positively associated with nursing frequency, and negatively with the proportion of nutritive nursings and baseline cortisol values. The results indicate that most aspects of pig maternal behaviour have not been significantly changed by domestication and that substantial variability in maternal behaviour exists between sows, perhaps in the form of several behaviour characteristics which encompass both behaviour and endocrine profiles of the sows.     

Between- and within-herd variation in blood and milk biomarkers in Holstein cows in early lactation

Both blood- and milk-based biomarkers have been analysed for decades in research settings, although often only in one herd, and without focus on the variation in the biomarkers that are specifically related to herd or diet. Biomarkers can be used to detect physiological imbalance and disease risk and may have a role in precision livestock farming (PLF). For use in PLF, it is important to quantify normal variation in specific biomarkers and the source of this variation. The objective of this study was to estimate the between- and within-herd variation in a number of blood metabolites (β-hydroxybutyrate (BHB), non-esterified fatty acids, glucose and serum IGF-1), milk metabolites (free glucose, glucose-6-phosphate, urea, isocitrate, BHB and uric acid), milk enzymes (lactate dehydrogenase and N-acetyl-β-D-glucosaminidase (NAGase)) and composite indicators for metabolic imbalances (Physiological Imbalance-index and energy balance), to help facilitate their adoption within PLF. Blood and milk were sampled from 234 Holstein dairy cows from 6 experimental herds, each in a different European country, and offered a total of 10 different diets. Blood was sampled on 2 occasions at approximately 14 days-in-milk (DIM) and 35 DIM. Milk samples were collected twice weekly (in total 2750 samples) from DIM 1 to 50. Multilevel random regression models were used to estimate the variance components and to calculate the intraclass correlations (ICCs). The ICCs for the milk metabolites, when adjusted for parity and DIM at sampling, demonstrated that between 12% (glucose-6-phosphate) and 46% (urea) of the variation in the metabolites’ levels could be associated with the herd-diet combination. Intraclass Correlations related to the herd-diet combination were generally higher for blood metabolites, from 17% (cholesterol) to approximately 46% (BHB and urea). The high ICCs for urea suggest that this biomarker can be used for monitoring on herd level. The low variance within cow for NAGase indicates that few samples would be needed to describe the status and potentially a general reference value could be used. The low ICC for most of the biomarkers and larger within cow variation emphasises that multiple samples would be needed - most likely on the individual cows - for making the biomarkers useful for monitoring. The majority of biomarkers were influenced by parity and DIM which indicate that these should be accounted for if the biomarker should be used for monitoring.     

Lack of stimulation of relaxin secretion in lactating sows by suckling in vivo or by oxytocin in vitro.

After parturition, eight sows were zero weaned by removing all piglets 6 h after birth; a further 18 sows suckled at least ten piglets each. Blood samples were collected on Day 4 after zero weaning or on Days 4, 14 and 21 of lactation and the sampling frequency increased during suckling bouts. Ovaries were recovered from sows on these days and corpora lutea were either extracted for estimation of relaxin and progesterone concentration, fixed for immunohistochemical analysis or incubated in vitro in the presence or absence of luteinizing hormone (LH) or oxytocin. Luteal weight and progesterone were higher in the zero-weaned sows than in lactating sows (P less than 0.05 and less than 0.001, respectively); relaxin content was below detection by Day 14. This was supported by immunohistochemical staining for relaxin, which showed limited immunostaining in zero-weaned and Day 4 sows, but none in the tissue recovered on Days 14 and 21, which showed typical signs of regression. Secretion of progesterone and relaxin by luteal tissue in vitro was highest in zero-weaned sows (P less than 0.05), decreased as lactation progressed and neither LH nor oxytocin had any significant effect. Concentrations of plasma relaxin were all less than 0.2 ng/ml in three of the four zero-weaned and Day-4-suckled sows assayed; there was no detectable increase during suckling bouts. It was concluded that during lactation the old corpus luteum of pregnancy is not able to release relaxin in response to suckling in vivo or to oxytocin treatment in vitro.     

Hormones, IgG and lactose changes around parturition in plasma, and colostrum or saliva of multiparous sows

Blood, colostrum and saliva samples were serially taken from 6 multiparous sows from day 109 of gestation until day 3 postpartum. Plasma was assayed for oestradiol-17beta (E2), progesterone (P4), prolactin (PRL), cortisol, immunoglobulin G (IgG) and lactose. Colostrum was assayed for E2, P4, IgG and lactose. Lactoserum, obtained after ultra centrifugation of colostrum, was assayed for PRL. Saliva was assayed for cortisol. Time-related variations in hormone, IgG and lactose concentrations measured in plasma were parallel to those measured in colostrum, lactoserum or saliva. However, the concentrations were higher in colostrum or lactoserum and lower in saliva than in plasma. Ratios of concentrations of cortisol in saliva and PRL in lactoserum over those in plasma did not vary with time and averaged 0.2 and 1.6, respectively. Conversely, the ratios of concentrations of E2 and P4 in colostrum over those in plasma varied with time (P < 0.05) but were quite constant before the end of parturition, averaging 2.7 and 3.6, respectively. The ratios of concentrations of IgG and lactose in colostrum over those in plasma also varied with time (P < 0.05). The concentrations of hormones in plasma on the one hand and in colostrum, lactoserum or saliva on the other hand were significantly correlated but correlations varied with time (PRL across periods: r = 0.31; cortisol across periods: r = 0.60; E2 during parturition: r = 0.83; P4 before parturition: r = 0.82; P4 during parturition: r = 0.67). The present results indicate that around parturition, assays of hormones in colostrum or saliva can be used to study the hormonal status of sows. Furthermore, variations in colostrum and plasma concentrations of IgG and lactose are good indicators of the transition from colostrum to milk synthesis.     

Optimizing Human Synovial Fluid Preparation for Two-Dimensional Gel Electrophoresis

Background

Prenatal screening for Down Syndrome (DS) would benefit from an increased number of biomarkers to improve sensitivity and specificity. Improving sensitivity and specificity would decrease the need for potentially risky invasive diagnostic procedures.

Results

We have performed an in depth two-dimensional difference gel electrophoresis (2D DIGE) study to identify potential biomarkers. We have used maternal plasma samples obtained from first and second trimesters from mothers carrying DS affected fetuses compared with mothers carrying normal fetuses. Plasma samples were albumin/IgG depleted and expanded pH ranges of pH 4.5 - 5.5, pH 5.3 - 6.5 and pH 6 - 9 were used for two-dimensional gel electrophoresis (2DE). We found no differentially expressed proteins in the first trimester between the two groups. Significant up-regulation of ceruloplasmin, inter-alpha-trypsin inhibitor heavy chain H4, complement proteins C1s subcomponent, C4-A, C5, and C9 and kininogen 1 were detected in the second trimester in maternal plasma samples where a DS affected fetus was being carried. However, ceruloplasmin could not be confirmed as being consistently up-regulated in DS affected pregnancies by Western blotting.

Conclusions

Despite the in depth 2DE approach used in this study the results underline the deficiencies of gel-based proteomics for detection of plasma biomarkers. Gel-free approaches may be more productive to increase the number of plasma biomarkers for DS for non-invasive prenatal screening and diagnosis.     

Increased dietary protein for lactating sows affects body composition,blood metabolites and milk production

Hyper-prolific sows nurse more piglets than less productive sows, putting a high demand on the nutrient supply for milk production. In addition, the high production level can increase mobilization from body tissues. The effect of increased dietary protein (104, 113, 121, 129, 139 and 150 g standardized ileal digestible (SID) CP/kg) on sow body composition, milk production and plasma metabolite concentrations was investigated from litter standardization (day 2) until weaning (day 24). Sow body composition was determined using the deuterium oxide dilution technique on days 3 and 24 postpartum. Blood samples were collected weekly, and milk samples were obtained on days 3, 10 and 17 of lactation. Litter average daily gain (ADG) peaked at 135 g SID CP/kg (P < 0.001). Sow BW and back fat loss reached a breakpoint at 143 and 127 g SID CP/kg (P < 0.001). Milk fat increased linearly with increasing dietary SID CP (P < 0.05), and milk lactose decreased until a breakpoint at 124 g SID CP/kg and 5.3% (P < 0.001) on day 17. The concentration of milk protein on day 17 increased until a breakpoint at 136 g SID CP/kg (5.0%; P < 0.001). The loss of body protein from day 3 until weaning decreased with increased dietary SID CP until it reached a breakpoint at 128 g SID CP/kg (P < 0.001). The body ash loss declined linearly with increasing dietary SID CP (P < 0.01), and the change in body fat was unaffected by dietary treatment (P=0.41). In early lactation (day 3 + day 10), plasma urea N (PUN) increased linearly after the breakpoint at 139 g SID CP/kg at a concentration of 3.8 mmol/l, and in late lactation (day 17 + day 24), PUN increased linearly after a breakpoint at 133 g SID CP/kg (P < 0.001) at a concentration of 4.5 mmol/l. In conclusion, the SID CP requirement for sows was estimated to 135 g/kg based on litter ADG, and this was supported by the breakpoints of other response variables within the interval 124 to 143 g/kg.     

Concentrations and origin of oxytocin in breast milk

Samples of human milk obtained from lactating women in the early postpartum period were assayed for oxytocin concentrations by specific RIA, following extraction procedures with Florisil. Mean oxytocin concentrations in human milk at postpartum day 1 to 5 were 4.5 +/- 1.1, 4.7 +/- 1.1, 4.0 +/- 1.3, 3.2 +/- 0.4, 3.3 +/- 0.6 microunits/ml (+/- SE), respectively. Oxytocin levels in milk were significantly increased by nursing (3.1 +/- 0.6, 5.3 +/- 1.0 microunits/ml, respectively). 3H-oxytocin in human milk was stable even after incubation at 37 degrees C for 2 hours. The dilution curve for milk was parallel to the curve for the standard oxytocin. The chromatographic fraction of immunoreactive oxytocin was identical to that of 3H-oxytocin. 3H-oxytocin was administered to lactating rats. Radioactivity in the neonatal gastric contents and plasma were 12.8% and 4.4% of the counts in the maternal plasma. It was made clear that oxytocin is stable in milk and that oxytocin in maternal blood can be transferred to mik and then to neonates.     

Circulating tumour necrosis factor-α bioactivity in rheumatoid arthritis patients treated with infliximab: link to clinical response

Our objective was to clarify the heterogeneity in response to infliximab treatment in rheumatoid arthritis (RA); to this end, a bioassay was designed to explore the contribution of circulating tumour necrosis factor (TNF)-α bioactivity and its possible link to response. The bioassay is based on the induction of IL-6 and osteoprotegerin (OPG) production by synoviocytes in response to TNF-α. RA synoviocytes were cultured with TNF-α (5 ng/ml) and 42 RA plasma samples collected just before starting therapy. Levels of IL-6 and OPG were measured in supernatants. In 20 of the patients, plasma samples collected before and 4 hours after the first and the ninth infusions were tested in the same way. Plasma concentrations of TNF-α and p55 and p75 soluble receptors were measured using ELISA. TNF-α induced IL-6 and OPG production by synoviocytes, which was further increased with patient plasma dilutions and inhibited by infliximab. With plasma samples obtained before the first infusion, the IL-6-induced production was greater in patients with a good clinical response than in the poor responders (44.4 ± 23.3 ng/ml versus 27.4 ± 20.9 ng/ml; P = 0.05). This high circulating TNF-α bioactivity was strongly inhibited with the first infliximab infusion. The difference between IL-6 levels induced with plasma samples obtained before and 4 hours after the first infusion was greater in patients with a good clinical response (40.0 ± 23.7 ng/ml versus 3.4 ± 10.0 ng/ml; P = 0.001). Similar findings were obtained for OPG production (7.0 ± 6.2 ng/ml versus 0.0 ± 3.0 ng/ml; P < 0.05). Levels of circulating TNF-α bioactivity were predictive of clinical response to TNF-α inhibition, confirming a key role for TNF-α in these RA patients.     

Metabolic fingerprinting and fermentative capacity of the intestinal flora of pigs during pre- and post-weaning periods

We used the Phene Plate generalized microplates to investigate the metabolic activities of faecal flora of pigs during pre- and post-weaning periods. Weekly samples were collected from four sows and their litters (four piglets from each) during 5 months. The metabolic fingerprints obtained from faecal floras of sows and their litters in the first sampling occasion was very similar, suggesting that sows were the initial source of flora for piglets. This similarity, however, was lost in week 2 and piglets developed new types of flora which, although similar among the litter-mates, differed from those of the sows. The metabolic fingerprints of pigs' floras during the post-weaning period also differed from those of the suckling period. On day 70 pigs were transferred to a fattening stable. The faecal flora of the animals during this period was unstable in each individual and differed among litter-mates. A pattern of successive changes was observed in the fermentative capacity (FC) of pigs' floras reaching the highest value before weaning (day 34). An overall decrease in the FC value of faecal floras was observed as a consequence of dietary shifts from milk to solid food to high energy fattening diet. The mean FC value of pig floras on the first sampling occasion (mean ± S.D. = 0·41 ± 0·02) was significantly higher than that obtained in the last sampling occasion (day 145) (0·31 ± 0·04) ( P < 0·001). Faecal floras of sows also had lower FC values (0·25 ± 0·01) than those of piglets suggesting that loss of FC by the faecal flora of young pigs will continue as they age.     

Identification of promising plasma immune biomarkers to differentiate active pulmonary tuberculosis

Although much research has been done related to biomarker discovery for tuberculosis infection, a set of biomarkers that can discriminate between active and latent TB diseases remains elusive. In the current study we correlate clinical aspects of TB disease with changes in the immune response as determined by biomarkers detected in plasma. Our study measured 18 molecules in human plasma in 17 patients with active disease (APTB), 14 individuals with latent tuberculosis infection (LTBI) and 16 uninfected controls (CTRL). We found that active tuberculosis patients have increased plasma levels of IL-6, IP-10, TNF-α, sCD163 and sCD14. Statistical analysis of these biomarkers indicated that simultaneous measurement of sCD14 and IL-6 was able to diagnose active tuberculosis infection with 83% accuracy. We also demonstrated that TNF-α and sCD163 were correlated with tuberculosis severity. We showed that the simultaneous detection of both plasma sCD14 and IL-6 is a promising diagnostic approach to identify APTB, and further, measurement of TNF-α and sCD163 can identify the most severe cases of tuberculosis.