Genome-wide methylation profile following prenatal and postnatal dietary omega-3 fatty acid supplementation in pigs.

The objective of this study was to determine how prenatal and postnatal dietary omega-3 fatty acids alter white blood cell (leukocyte) DNA methylation of offspring. Fifteen gilts (n = 5 per treatment) were selected from one of three treatments: (i) control diet throughout gestation, lactation and nursery phase (CON); (ii) algal omega-3 fatty acid supplementation enriched in EPA and DHA (Gromega™ ) fed throughout gestation, lactation and nursery phase (Cn3); or (iii) Gromega™ supplementation maternally, during gestation and lactation only, and control diet during the nursery phase (Mn3). At 11 weeks of age and after 8 weeks of post-weaning nursery feeding, buffy coat genomic DNA was subjected to methyl CpG binding protein sequencing. The methylation enriched profile mapped to 26% of the porcine genome. On chromosome 4, a 27.7-kb differentially methylated region downstream of RUNX1T1 was hypomethylated in the Mn3 and Cn3 groups by 91.6% and 85.0% respectively compared to CON pigs. Conversely, hypermethylation was detected in intergenic regions of chromosomes 4 and 12. Regulatory impact factor and differential hubbing methods were used to identify pathways that were coordinately regulated by methylation due to feeding EPA and DHA during pregnancy. Despite limited ability to detect differential methylation, we describe methods that allow the identification of coordinated epigenetic regulation that could not otherwise be detected from subtle single locus changes in methylation. These data provide evidence of novel epigenetic regulation by maternal and early life supplementation of omega-3 fatty acids that may have implications to growth and inflammatory processes.

Effects of dietary enrichment with a marine oil-based n-3 LCPUFA supplement in sows with predicted birth weight phenotypes on growth performance and carcass quality of offspring.

Effects of a marine oil-based n-3 LCPUFA supplement (mLCPUFA) fed from weaning until the end of the next lactation to sows with a predicted low litter birth weight (LBW) phenotype on growth performance and carcass quality of litters born to these sows were studied, based on the hypothesis that LBW litters would benefit most from mLCPUFA supplementation. Sows were allocated to be fed either standard corn/soybean meal-based gestation and lactation diets (CON), or the same diets enriched with 0.5% of the mLCPUFA supplement at the expense of corn. The growth performance from birth until slaughter of the litters with the lowest average birth weight in each treatment (n=24 per treatment) is reported in this paper. At weaning, each litter was split between two nursery pens with three to six pigs per pen. At the end of the 5-week nursery period, two barrows and two gilts from each litter that had individual birth weights closest to their litter average birth weight, were moved to experimental grow-finish pens (barn A), where they were housed as two pigs per pen, sorted by sex within litter. Remaining pigs in each litter were moved to another grow-finish barn (barn B) and kept in mixed-sex pens of up to 10 littermates. After 8 weeks, one of the two pigs in each pen in barn A was relocated to the pens holding their respective littermates in barn B. The remaining barrows and gilts were individually housed in the pens in barn A until slaughter. Maternal mLCPUFA supplementation increased docosahexaenoic acid (DHA) concentration in the brain, liver and Semitendinosus muscle of stillborn pigs (P<0.01), did not affect eicosapentaenoic acid and DHA concentrations in sow serum at the end of lactation, and did not affect average daily gain, average daily feed intake or feed utilization efficiency of the offspring. BW was higher (P<0.01) in the second half of the grow-finish phase in pigs from mLCPUFA sows compared with controls in barn A, where space and competition for feed was minimal, but not barn B. Carcass quality was not affected by treatment for pigs from barn A, but maternal mLCPUFA supplementation negatively affected carcass quality in pigs from barn B. Collectively, these results suggest that nutritional supplementation of sows can have lasting effects on litter development, but that feeding mLCPUFA to sows during gestation and lactation was not effective in improving growth rates or carcass quality of LBW litters.

Effects of dietary enrichment with a marine oil-based n-3 LCPUFA supplement in sows with predicted birth weight phenotypes on birth litter quality and growth performance to weaning.

The effects of a marine oil-based n-3 long-chain polyunsaturated fatty acid (mLCPUFA) supplement fed to the sow from weaning, through the rebreeding period, during gestation and until end of lactation on litter characteristics from birth until weaning were studied in sows with known litter birth weight phenotypes. It was hypothesized that low birth weight (LBW) litters would benefit more from mLCPUFA supplementation than high birth weight litters. A total of 163 sows (mean parity=4.9 ± 0.9) were rebred after weaning. Sows were pair-matched by parity and litter average birth weight of the previous three litters. Within pairs, sows were allocated to be fed either standard corn/soyabean meal-based gestation and lactation diets (CON), or the same diets enriched with 0.5% of the mLCPUFA supplement at the expense of corn. Each litter between 9 and 16 total pigs born was classified as LBW or medium/high average birth weight (MHBW) litter and there was a significant correlation (P<0.001) between litter average birth weight of the current and previous litters within sows (r=0.49). Sow serum was harvested at day 113 of gestation for determination of immunoglobulin G (IgG) concentrations. The number of pigs born total and alive were lower (P=0.01) in mLCPUFA than CON sows, whereas the number of stillborn and mummified pigs were similar between treatments. Number of stillborns (trend) and mummies (P<0.01) were higher in LBW than MHBW litters. Tissue weights and brain : tissue weight ratios were similar between treatments, but LBW litters had decreased tissue weights and increased brain : tissue weight ratios compared with MHBW litters. Placental weight was lower (P=0.01) in LBW than MHBW litters, but was not different between treatments. Average and total litter weight at day 1 was similar between treatments. mLCPUFA increased weaning weight (P=0.08) and average daily gain (P<0.05) in MHBW litters, but not in LBW litters. Pre-weaning mortality was similar between treatments, but was higher (P<0.01) in LBW than MHBW litters. IgG concentration in sow serum was similar between treatments and litter birth weight categories. In conclusion, litter birth weight phenotype was repeatable within sows and LBW litters showed the benchmarks of intra-uterine growth retardation (lower placental weight and brain sparing effects). As maternal mLCPUFA supplementation decreased litter size overall, only improved litter growth rate until weaning in MHBW litters, and did not affect pre-weaning mortality, maternal mLCPUFA supplementation was not an effective strategy in our study for mitigating negative effects of a LBW litter phenotype.

The impact of dietary long-chain polyunsaturated fatty acids on bone and cartilage in gilts and sows.

Dietary long-chain PFO including arachidonic acid (ARA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) are precursors for several inflammatory mediators. The objective of this study was to characterize the effects of dietary PFO supplementation on bone, cartilage, and synovial fluid in 2 ages of pigs. Sows and gilts were fed either control corn/soybean meal based diets or the control diet supplemented with PFO from Gromega (PFO; JBS United, Sheridan, IN). Sows were fed their diets for 24.5 mo and slaughtered at 43 mo while gilts were fed their diets from weaning until slaughter at 111 kg. Cartilage was harvested from both humeroulnar joints of 14 sows (7/treatment) and 16 gilts (8/treatment) within 30 h of slaughter for fatty acid analysis and explant cultures. Synovial fluid was collected from the carpal joints of each pig postmortem. The right fused radius/ulna was collected for computed tomography (CT) analysis. Cortical width and density were determined and trabecular density was measured at the distal radius. Cartilage explants were allocated to 24-well culture plates with 2 discs per well and cultured over 72 h at 37°C in serum-free Dulbecco's modified Eagle's medium: nutrient mixture F-12 (Ham) medium. Six wells/pig were treated with 10 ng/mL of recombinant porcine interleukin-1 (rpIL-1). At 24, 48, and 72 h of culture, media were removed and reserved for analysis of proteoglycans, nitric oxide (NO), and PGE2 concentrations. The CT scans of the radius/ulna from gilts revealed no differences for cortical width and bone density. Sows fed PFO had greater cortical width of the proximal ulna (P < 0.05) and decreased cortical width of the distal radius (P < 0.05). Sows fed PFO had increased DHA (P < 0.01) and a decrease in the omega-6 to omega-3 ratio (P < 0.05) in cartilage. Gilts fed PFO had increased DHA (P < 0.01), C22:1 (P < 0.01), and docosapentaenoic acid (P < 0.01) and a tendency for increased EPA (P = 0.093) concentrations in cartilage. Changes in dietary fatty acids in the gilts and sows had no effect on the variables tested in vitro. Although the PFO diet increased omega-3 incorporation into chondrocytes, the biological significance is unclear since concentrations of ARA were at least 9-fold higher than EPA or DHA. Therefore, if omega-3 fatty acids can mitigate inflammation in joints, the benefit would likely either be the result of systemic changes in inflammatory mediators or higher concentrations in the diet.

Consequences of a low litter birth weight phenotype for postnatal lean growth performance and neonatal testicular morphology in the pig.

The consequences of a low litter average birth weight phenotype for postnatal growth performance and carcass quality of all progeny, and testicular development in male offspring, were investigated. Using data from 25 sows with one, and 223 sows with two consecutive farrowing events, individual birth weight (BW) was measured and each litter between 9 and 16 total pigs born was classified as low (LBW), medium (MBW) or high (HBW) birth weight: low and high BW being defined as >1 standard deviation below or above, respectively, the population mean for each litter size. Litter average BW was repeatable within sows. At castration, testicular tissue was collected from 40 male pigs in LBW and HBW litters with individual BW close to their litter average BW and used for histomorphometric analysis. LBW piglets had a lower absolute number of germ cells, Sertoli cells and Leydig cells in their testes and a higher brain : testis weight ratio than HBW piglets. Overall, LBW litters had lower placental weight and higher brain : liver, brain : intestine and brain : Semitendinosus muscle weight ratios than MBW and HBW litters. In the nursery and grow-finish (GF) phase, pigs were kept in pens by BW classification (9 HBW, 17 MBW and 10 LBW pens) with 13 males and 13 females per pen. Average daily gain tended to be lower in LBW than HBW litters in lactation (P = 0.06) and throughout the nursery and GF phases (P < 0.01), resulting in an increasing difference in body weight between LBW, MBW and HBW litters (P < 0.05). Average daily feed intake was lower (P < 0.001) in LBW than HBW litters in the nursery and GF phases. Feed utilization efficiency (feed/gain) was similar for LBW and HBW litters in the nursery, but was lower (P < 0.001) in HBW than LBW litters in the GF phase. By design, slaughter weight was similar between BW classifications; however, LBW litters needed 9 more days to reach the same slaughter weight than HBW litters (P < 0.001). BW classification did not affect carcass composition traits. In conclusion, LBW litters showed benchmarks of intrauterine growth retardation, LBW had a negative impact on testicular development and germ and somatic cell populations, and was associated with decreased postnatal growth during all phases of production; however, no measurable effect on carcass composition traits was established.

The effect of a Bacillus-based direct-fed microbial supplemented to sows on the gastrointestinal microbiota of their neonatal piglets.

Direct-fed microbials (DFM) supplemented in sow diets may confer health benefits to the host and their piglets by reducing pathogens in the sow and environment. In this study we evaluated the effect of a Bacillus-based DFM on the gastrointestinal microbiota of neonatal piglets. A total of 208 sows were divided into 2 treatments: a control diet and the control diet supplemented with a Bacillus subtilis-based DFM (3.75 × 10(5) cfu/g feed). Twenty-one piglets sampled from each sow treatment group were euthanized on d 3 of lactation followed by an additional 15 piglets per treatment on d 10 of lactation. Litters from DFM-supplemented sows had greater (P = 0.02) weaning weights and a tendency (P = 0.09) for improvement in litter ADG. Sows supplemented with the DFM weaned more pigs (P = 0.06) than control sows which was reflected in numerically lower but not statistically different (P = 0.12) decrease in piglet mortality in DFM litters. Terminal RFLP was used to characterize gastrointestinal (GI) microbial populations in the ileum and colon of the piglets. Terminal restriction fragments (T-RF) were compared between control and DFM treatments. There was a greater incidence and quantity of T-RF B423 and H330 (binary P = 0.01, 0.08; quantitative P = 0.01, 0.05, respectively), putatively identified as Lactobacillus gasseri/johnsonii, in the ileum of pigs nursing sows supplemented with DFM at d 3. Terminal restriction fragment peaks B423 and H330 were also greater (binary P = 0.01, 0.08; quantitative P = 0.01, 0.01, respectively) in the colon of pigs nursing sows supplemented with DFM at d 3. Peaks M495 and B394, putatively identified as E. coli, were greater (binary P = 0.01, 0.04; quantitative P = 0.01, 0.01, respectively) in the colon of the control pigs at d 3. At d 10, both the presence and quantity of Lactobacillus species were greater (P < 0.05) in the colon of pigs with the DFM treatment. Additionally, there was a tendency for T-RF B227 and H257 (binary P = 0.07, 0.07, respectively), putatively identified as Clostridium perfringens, to be present in the ileum of the control pigs at d 10 compared with treated pigs. Results of this study reveal that the developing gastrointestinal microbiota of a neonatal piglet can be affected by DFM supplementation to the sow.

Responses to n-3 fatty acid (LCPUFA) supplementation of gestating gilts, and lactating and weaned sows.

Feeding n-3 long-chain polyunsaturated fatty acids (LCPUFA) to gilts or sows has shown different responses to litter growth, pre-weaning mortality and subsequent reproductive performance of the sow. Two hypotheses were tested: (1) that feeding a marine oil-based supplement rich in protected n-3 LCPUFAs to gilts in established gestation would improve the growth performance of their litters; and (2) that continued feeding of the supplement during lactation and after weaning would offset the negative effects of lactational catabolism induced, using an established experimental model involving feed restriction of lactating primiparous sows. A total of 117 primiparous sows were pair-matched at day 60 of gestation by weight, and when possible, litter of origin, and were allocated to be either control sows (CON) fed standard gestation and lactation diets, or treated sows (LCPUFA) fed the standard diets supplemented with 84 g/day of a n-3 LCPUFA rich supplement, from day 60 of first gestation, through a 21-day lactation, and until euthanasia at day 30 of their second gestation. All sows were feed restricted during the last 7 days of lactation to induce catabolism, providing a background challenge against which to determine beneficial effects of n-3 LCPUFA supplementation on subsequent reproduction. In the absence of an effect on litter size or birth weight, n-3 LCPUFA tended to improve piglet BW gain from birth until 34 days after weaning (P = 0.06), while increasing pre-weaning mortality (P = 0.05). It did not affect energy utilization by the sow during lactation, thus not improving the catabolic state of the sows. Supplementation from weaning until day 30 of second gestation did not have an effect on embryonic weight, ovulation rate or early embryonic survival, but did increase corpora lutea (CL) weight (P = 0.001). Eicosapentaenoic acid and docosahexaenoic acid (DHA) levels were increased in sow serum and CL (P < 0.001), whereas only DHA levels increased in embryos (P < 0.01). In conclusion, feeding n-3 LCPUFA to gilts tended to improve litter growth, but did not have an effect on overall subsequent reproductive performance.

Senile hair graying: H2O2-mediated oxidative stress affects human hair color by blunting methionine sulfoxide repair.

Senile graying of human hair has been the subject of intense research since ancient times. Reactive oxygen species have been implicated in hair follicle melanocyte apoptosis and DNA damage. Here we show for the first time by FT-Raman spectroscopy in vivo that human gray/white scalp hair shafts accumulate hydrogen peroxide (H(2)O(2)) in millimolar concentrations. Moreover, we demonstrate almost absent catalase and methionine sulfoxide reductase A and B protein expression via immunofluorescence and Western blot in association with a functional loss of methionine sulfoxide (Met-S=O) repair in the entire gray hair follicle. Accordingly, Met-S=O formation of Met residues, including Met 374 in the active site of tyrosinase, the key enzyme in melanogenesis, limits enzyme functionality, as evidenced by FT-Raman spectroscopy, computer simulation, and enzyme kinetics, which leads to gradual loss of hair color. Notably, under in vitro conditions, Met oxidation can be prevented by L-methionine. In summary, our data feed the long-voiced, but insufficiently proven, concept of H(2)O(2)-induced oxidative damage in the entire human hair follicle, inclusive of the hair shaft, as a key element in senile hair graying, which does not exclusively affect follicle melanocytes. This new insight could open new strategies for intervention and reversal of the hair graying process.

Regulation of pigmentation in human epidermal melanocytes by functional high-affinity beta-melanocyte-stimulating hormone/melanocortin-4 receptor signaling.

To date, the principal receptor considered to regulate human pigmentation is the melanocortin-1 receptor (MC1-R) via induction of the cAMP/protein kinase A pathway by the melanocortins alpha-MSH and ACTH. In this context, it is noteworthy that beta-MSH can also induce melanogenesis, although it has a low affinity for the MC1-R, whereas the preferred receptor for this melanocortin is the MC4-R. Because beta-MSH is present in the epidermal compartment, it was of interest to ascertain whether functioning MC4-Rs are present in human epidermal keratinocytes and melanocytes. Our results provide evidence that the MC4-R is expressed in situ and in vitro throughout the human epidermis at the mRNA and protein level using RT-PCR, Western blotting, and double immunofluorescence staining. Moreover, radioligand binding studies yielded high-affinity receptors for beta-MSH on epidermal melanocytes (3600 receptors per cell), undifferentiated keratinocytes (7200 receptors per cell), and differentiated keratinocytes (72,700 receptors per cell), indicating that MC4-R expression correlates with epidermal differentiation. Importantly, increased melanogenesis after stimulation of the beta-MSH/cAMP/microphthalmia-associated transcription factor/tyrosinase cascade proved the functionality of this signal in melanocytes, which was attenuated in the presence of the specific MC4-R antagonist HS014. In summary, our results imply an important role for the beta-MSH/MC4-R cascade in human melanocyte biology, although the function and purpose of this signal in keratinocytes needs further elucidation.

The Ca2+-binding capacity of epidermal furin is disrupted by H2O2-mediated oxidation in vitiligo.

The Ca(2+)-dependent precursor convertase furin is abundantly expressed in epidermal keratinocytes and melanocytes. In this context, it is noteworthy that proopiomelanocortin (POMC) cleavage is also processed by furin, leading to ACTH, beta-lipotropin, and beta-endorphin. All prohormone convertases including furin are regulated by Ca(2+). Because numerous epidermal peptides and enzymes are affected by H(2)O(2)-mediated oxidation, including the POMC-derived peptides alpha-MSH and beta-endorphin as shown in the epidermis of patients with vitiligo, we here asked the question of whether furin could also be a possible target for this oxidation mechanism by using immunofluorescence, RT-PCR, Western blotting, Ca(2+)-binding studies, and computer modeling. Our results demonstrate significantly decreased in situ immunoreactivity of furin in the epidermis of patients with progressive vitiligo (n = 10), suggesting H(2)O(2)-mediated oxidation. This was confirmed by (45)Ca(2+)-binding studies with human recombinant furin identifying the loss of one Ca(2+)-binding site from the enzyme after oxidation with H(2)O(2). Computer simulation supported alteration of one of the two Ca(2+)-binding sites on furin. Taken together, our results implicate that the Ca(2+)-dependent proteolytic activity of this convertase is targeted by H(2)O(2), which in turn could contribute to the reduced epidermal expression of the POMC-derived peptides alpha-MSH and beta-endorphin as documented earlier in patients with vitiligo.

Diet modifications to improve finishing pig growth performance and pork quality attributes during periods of heat stress.

A total of 196 barrows (88 kg) were used in a 2 x 2 factorial arrangement of treatments and housed in a facility (seven pigs per pen) where temperatures cycled between 27 and 35 degrees C. Treatments consisted of (as-fed basis) two CP levels (13.6 or 11.3%) and two levels of added fat (1 or 8%). Diets were formulated to the same true digestible lysine:ME ratio (1.68 g of lysine/Mcal of ME). Diets were fed and growth variables were measured until pigs reached 114 kg of BW. Ham and LM (loin) 24-h pH (PH24), and light reflectance (CIE L*, and a*, and b*, and hue angle) were taken after slaughter. Additionally, loins were removed and measured for i.m. fat, moisture, glycolytic potential, and subjected to a 7-d retail display evaluation that measured pH, light reflectance, and subjective color and odor score. The remaining boneless lumbar loin segment was vacuum-sealed for 14 d and subsequently measured for pH, light reflectance, and color. Pigs fed the high-CP, low-fat diet had a lower ADG than all other treatments (P = 0.06). High-fat feeding resulted in improved ADG (CP x Fat; P = 0.06) and G:F (Fat effect; P < 0.01). Higher fat and lower protein levels both increased final BF (P = 0.07). Pigs fed the low-CP diets had lower ham PH24 (P < 0.01). Loin PH24 was higher with high fat feeding (P = 0.10). Additionally, pigs fed high fat diets had lower L* values on the ham face and cut loin 24 h after slaughter (Fat effect; P

The 'true' acetabular index in children with cerebral palsy.

INTRODUCTION: The acetabular index is a measurement used to determine whether or not an acetabular osteotomy may be required at the time of hip reconstruction in children. METHODS: A retrospective analysis of pre- and postoperative values of the acetabular index was carried out on two groups of children. One group of children had an acetabuloplasty and the other did not during hip reconstruction, the decision being made at the time of open reduction, based on lack of hip congruity. RESULTS: When the pre-operative acetabular index measurements were reviewed, the mean difference between the two groups was only 7 degrees. Range of measurements was similar between the two groups. CONCLUSIONS: The acetabular index is difficult to measure in some children with cerebral palsy. The bony landmarks are difficult to standardise. This may explain the similar pre-operative values between the two groups of children in this study. We, therefore, recommend that before hip reconstruction in children with cerebral palsy, the measurement of the acetabular index should be abandoned. Decisions as to whether to undertake an acetabuloplasty or not should be made at the time of open reduction.

Pharmacological zinc levels reduce the phosphorus-releasing efficacy of phytase in young pigs and chickens.

A pig trial and a chick trial were done to determine the effect of high levels of Zn and Cu on the P-releasing efficacy of phytase. Ninety-nine individually fed pigs (7.2 kg) were given ad libitum access to one of 11 experimental diets for a period of 21 d. Fibula ash (mg) was regressed against supplemental inorganic P (iP) intake (g) to establish the standard curve, from which phytase treatments were compared to determine P-releasing efficacy. The basal diet was a corn-soybean meal diet with no supplemental P (21% CP, 0.075% estimated available P, 130 mg of Zn/kg, as-fed basis). Diets included three graded levels of supplemental iP (0, 0.075, 0.150%) from reagent-grade KH2PO4, two levels of phytase (500 and 1,000 FTU/kg) from EcoPhos, 1,500 mg of Zn/kg from either Waelz ZnO or basic Zn chloride (Zn5Cl2(OH)8), and all combinations of phytase and Zn. One phytase unit (FTU) was defined as the amount of enzyme required to release 1 micromol of iP per minute from sodium phytate at 37 degrees C and pH 5.5. Phytase supplementation improved (P < 0.01) weight gain, G:F, and fibula ash (% and mg). Bone ash (mg) was highest (P < 0.01) for pigs fed diets containing 1,000 FTU/kg of phytase. Supplemental Zn had no effect (P > 0.50) on growth performance, but decreased (P < 0.05) fibula ash (mg). Comparison of the phytase treatments to the standard curve (r2 = 0.87) revealed P-release values of 0.130 and 0.195% for 500 and 1,000 FTU of phytase/kg, respectively, in the absence of Zn, whereas in the presence of Zn (pooled), P-release values were decreased (P < 0.01) to 0.092 and 0.132%, respectively. The effects of high levels of supplemental Zn (basic Zn chloride) and Cu (CuSO4 x 5H2O) on phytase efficacy also were investigated in a 12-d chick trial. Dietary treatments were arranged according to a 2(3) factorial, with two levels each of supplemental phytase (0 and 500 FTU/kg from EcoPhos), Zn (0 and 800 mg/kg), and Cu (0 and 200 mg/kg). There was a phytase x Zn interaction (P < 0.01) for tibia ash. Thus, Zn supplementation decreased tibia ash in the presence, but not in the absence, of phytase. Supplemental Cu did not affect (P > 0.30) the response to phytase. These results suggest that pharmacological levels of Zn chelate the phytate complex, thereby decreasing its availability for hydrolysis by phytase.

Early weaning to reduce tissue mobilization in lactating sows and milk supplementation to enhance pig weaning weight during extreme heat stress.

This study was conducted to determine the effect of reduced lactation length and supplemental milk replacer (MR) during high ambient temperatures. Thirty nine primiparous and 100 multiparous sows (PIC, Franklin, KY, C-22) were used in a 2 x 2 x 2 factorial arrangement of treatments. Treatments consisted of two lactation room temperatures (21 degrees C [TN] and 32 degrees C [HOT]), two lactation lengths (14 or 19 d), and two parity groups (primiparous, multiparous). Pigs were either: 1) sow-reared to 19 d or 2) sow-reared to 14 d, and then reared to 19 d with MR after sow removal. All sows were fed the same diet (1.07% lysine, 3,366 kcal of ME/kg). Sows were weighed and ultrasound for backfat thickness (BF) and longissimus muscle area (LMA) within 6 h after farrowing and at the time of sow removal (d 14 or 19). Pigs were individually weighed at weaning (d 19) and after a 47-d nursery period (d 66). Heat stress increased sow weight loss (-13.35 kg, P < 0.01) and decreased sow feed intake (4.63 kg/d, P < 0.01) during lactation compared with sows in TN (+4.5 kg and 7.5 kg/d, respectively). Early weaning (d 14) during heat stress decreased maternal weight loss (-10.1 vs. -16.6 kg, P < 0.01). Primiparous sows lost more BF in both environments (-2.60 vs. -1.56 mm, P < 0.05), and both parity groups lost more BF (-3.35 vs. -2.3 mm, P < 0.10) and LMA (-1.82 vs. -0.77 cm2, P < 0.05) when lactating for 19 d in the HOT environment than those lactating for 14 d. Pigs nursing primiparous and multiparous sows in the HOT environment and provided MR had heavier individual 19-d weights (7.37 and 8.12 kg/ pig, respectively) than those nursing to 19 d (5.57 and 6.04 kg/pig, P < 0.01). Milk replacer decreased the difference normally observed in 19-d weights between primiparous and multiparous sow-reared pigs in TN. Pigs fed MR in both environments and nursing multiparous sows had improved weight gains in the nursery compared with pigs nursing sows to 19 d (428 vs. 406 g/d, respectively; P < 0.01), or reared by primiparous sows (444 vs. 390 g/d , respectively; P < 0.01). Sow weaning on d 14 in the HOT environment decreased the wean-to-estrus interval in primiparous sows (22.8 vs. 9.2 d, P < 0.10). This study shows the benefit of early weaning in combination with milk replacer to preserve the sow and to restore pig weaning weights and nursery end weights under heat stress.

Effect of menhaden fish oil supplementation and lipopolysaccharide exposure on nursery pigs. I. Effects on the immune axis when fed diets containing spray-dried plasma.

The objective of the present study was to evaluate the potential immunological benefit of adding menhaden fish oil to the diet of weaned pigs. Twenty-four crossbred male pigs were weaned at approximately 18 days of age and placed on a complex nursery diet containing 30% lactose and 7% plasma protein with 6% corn oil as the fat source (Cont, n=12) or with 5% menhaden fish oil and 1% corn oil as the fat source (MFO, n=12) for a period of 15 days. Body weights did not differ (P>0.78) between dietary groups either at the beginning or end of the 15 days feeding period. On day 15, all pigs were non-surgically fitted with an indwelling jugular catheter. On d 16, pigs received an i.v. injection of either saline (n=6/dietary group) or lipopolysaccharide (LPS; 150 microg/kg body weight; n=6/dietary group) and blood samples were collected at 30 min intervals for a period of 5h. Serum was harvested and stored at -80 degrees C for analysis of cortisol (CS), corticosteroid-binding globulin (CBG), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). There was no significant effect of diet on basal concentrations (Time 0) of any of the blood parameters analyzed. A Time x Treatment x Diet interaction (P<0.03) was observed for serum CS such that those pigs which consumed the MFO diet followed by LPS treatment had a reduced CS response as compared to the LPS-treated pigs on the Cont diet. A Time x Treatment interaction (P<0.01) was observed for serum CBG such that LPS treatment reduced circulating CBG as compared to the saline-treated pigs. Time x Treatment x Diet interactions were also observed for serum concentrations of TNF-alpha (P=0.084) and IFN-gamma (P=0.022) such that both the TNF-alpha and IFN-gamma response to the LPS challenge was lower in those pigs receiving the MFO diet as compared to the LPS-treated pigs on the Cont diet. Overall, serum CS was negatively correlated with the CBG response (r=-0.40, P<0.001), however, the strongest negative correlation was observed in the LPS-treated pigs which consumed the MFO diet (r=-0.63, P<0.001). While further studies are needed to evaluate the immunological response of including MFO in the nursery pig diet, the present study demonstrates that supplementation with MFO does indeed alter the immunological response to an LPS challenge.

Murray Valley encephalitis virus surveillance and control initiatives in Australia. National Arbovirus Advisory Committee of the Communicable Diseases Network Australia.

Mechanisms for monitoring Murray Valley encephalitis (MVE) virus activity include surveillance of human cases, surveillance for activity in sentinel animals, monitoring of mosquito vectors and monitoring of weather conditions. The monitoring of human cases is only one possible trigger for public health action and the additional surveillance systems are used in concert to signal the risk of human disease, often before the appearance of human cases. Mosquito vector surveillance includes mosquito trapping for speciation and enumeration of mosquitoes to monitor population sizes and relative composition. Virus isolation from mosquitoes can also be undertaken. Monitoring of weather conditions and vector surveillance determines whether there is a potential for MVE activity to occur. Virus isolation from trapped mosquitoes is necessary to define whether MVE is actually present, but is difficult to deliver in a timely fashion in some jurisdictions. Monitoring of sentinel animals indicates whether MVE transmission to vertebrates is actually occurring. Meteorological surveillance can assist in the prediction of potential MVE virus activity by signalling conditions that have been associated with outbreaks of Murray Valley encephalitis in humans in the past. Predictive models of MVE virus activity for south-eastern Australia have been developed, but due to the infrequency of outbreaks, are yet to be demonstrated as useful for the forecasting of major outbreaks. Surveillance mechanisms vary across the jurisdictions. Surveillance of human disease occurs in all States and Territories by reporting of cases to health authorities. Sentinel flocks of chickens are maintained in 4 jurisdictions (Western Australia, the Northern Territory, Victoria and New South Wales) with collaborations between Western Australia and the Northern Territory. Mosquito monitoring complements the surveillance of sentinel animals in these jurisdictions. In addition, other mosquito monitoring programs exist in other States (including South Australia and Queensland). Public health control measures may include advice to the general public and mosquito management programs to reduce the numbers of both mosquito larvae and adult vectors. Strategic plans for public health action in the event of MVE virus activity are currently developed or being developed in New South Wales, the Northern Territory, South Australia, Western Australia and Victoria. A southern tri-State agreement exists between health departments of New South Wales, Victoria and South Australia and the Commonwealth Department of Health and Aged Care. All partners have agreed to co-operate and provide assistance in predicting and combatting outbreaks of mosquito-borne disease in south-eastern Australia. The newly formed National Arbovirus Advisory Committee is a working party providing advice to the Communicable Diseases Network Australia on arbovirus surveillance and control. Recommendations for further enhancement of national surveillance for Murray Valley encephalitis are described.

Risk of hepatitis C virus infection in multiply transfused premature neonates.

BACKGROUND: Reports from around the world indicate that multiply transfused patients are at increased risk of hepatitis C virus (HCV) infection, with reported rates of between 4% and 44%. Such reports are mostly of haematological and renal patients. As recipients of blood products in the newborn period, premature infants share this risk, but there is little information regarding their risk. AIM: To assess the risk of HCV infection in children who, as premature neonates, received multiple blood products prior to the introduction of screening of donated blood for HCV. METHODS: Premature infants born between January 1985 and January 1990 who had attended our high-risk follow-up clinic were selected on the basis of the number of transfusions of blood, platelets or fresh frozen plasma they received in the newborn period. Ethical approval to offer HCV testing to parents was obtained from the Central Sydney Area Health Service Ethics Review Committee. Parents of infants who received three or more transfusions were then contacted by mail with the approved letter explaining the study, and offered HCV testing. Detection of anti-HCV antibodies was undertaken using second, and later third generation enzyme immunoassay kits. Samples which were found to be 'indeterminate' were tested using a Wellcozyme HCV western blot assay (Murex Diagnostics Ltd, Datford, UK). Hepatitis C virus-ribonucleic acid (RNA) was detected using an 'in-house' polymerase chain reaction (PCR) assay. Alanine transaminase (ALT) was also measured, with values above 55 U/L considered abnormal. RESULTS: Consent was obtained for 45 children (25 males, 20 females). The mean (+/- SEM) gestational age and weight of the children at birth was 26.7 +/- 0.2 weeks and 938 +/- 27 g, respectively. The children received 198 transfusions of blood products, an average of 4.4 U per child. All of the infants except for one were negative for anti-HCV antibodies. One infant was 'indeterminate' (low positive on third generation test but negative on second generation test), but proved negative subsequently on both western blot and PCR testing. HCV-RNA was not detected in any of the infants on PCR testing. All of the samples had normal ALT values, the mean being 16 U/L (range 8-52). CONCLUSION: None of the children consenting to this study had evidence of current HCV infection. Because of the sample size, we were not able to estimate the true risk of infection from this study, except that the upper limit for the risk is about 1/200 per transfused blood sample.

Langerhans cells in Dupuytren's contracture.

We have examined biopsies of Dupuytren's contracture palmar fascia, overlying subcutis and skin, and have correlated the distribution of gross macroscopic changes in the hand, mapped pre- and intraoperatively, with light microscopic immunohistochemical findings. We report increased numbers of S100 positive Langerhans cells (an epidermal cell of dendritic lineage) and CD45 positive cells, both in "nodules" and at dermo-epidermal junctions, in the biopsied tissues. This suggests that Langerhans cells migrate from the epidermis into Dupuytren's contracture tissue, possibly in response to local changes in levels of inflammatory cytokines within the tissue. Our findings, together with other reports of increased numbers of dermal dendrocytes and inflammatory cells in Dupuytren's contracture tissue, lend circumstantial support to the "extrinsic theory" of the pathogenesis of Dupuytren's contracture. However, the earliest stages of the disease process have not been defined, and therefore the events which ultimately produce fibrosis in the palmar fascial complex in susceptible individuals could begin in the skin and/or within deeper tissues, especially where there is dysregulation of the immune system.

Spontaneous clearance of hepatitis C virus infection post-liver transplantation is associated with rapidly changing quasispecies: a single case report.

Hepatitis C virus (HCV) clearance post-liver transplantation is uncommon. This is a case report of a patient who, after liver transplantation, developed cholestatic hepatitis characterized by severe graft dysfunction, in conjunction with high viral load. This was, however, followed by viral clearance and normalization of allograft function. The clinical features of this case and the quasispecies patterns during the illness and the clearance periods are described. In addition, management implications in terms of immunosuppressive therapy are discussed.

The Department of Defense Medical Mortality Registry.

The Department of Defense Medical Mortality Registry is being implemented at the Office of the Armed Forces Medical Examiner, Armed Forces Institute of Pathology, providing the first comprehensive medical mortality surveillance for the Department of Defense. The Registry attempts to obtain complete medical and circumstantial information on every military active duty death for medical surveillance and prevention research. Medical records, autopsy reports, eyewitness accounts, and investigative reports are reviewed to validate and synthesize medical, circumstantial, and risk factor information on each death. All military active duty deaths since 1980 are currently identified and classified by manner of death (accident, suicide, homicide, illness, hostile, undetermined). Military death rates have decreased during the past two decades by nearly half. About three-quarters of military deaths are attributable to injury (accident, suicide, homicide). The Registry creates new opportunities for prevention-oriented research as it collects detailed information on every military death.