Charting the proteome of Cryptosporidium parvum sporozoites using sequence similarity-based BLAST searching

Cryptosporidium (C.) spp. are important zoonotic parasites causing widespread diarrhoeal disease in man and animals. The recent release of the complete genome sequences for C. parvum and C. hominis has facilitated the comprehensive global proteome analysis of these opportunistic pathogens. The well-known approach for mass spectrometry (MS) based data analysis using the BLAST tool (MS BLAST) is a database search protocol for identifying unknown proteins by sequence similarity to homologous proteins using peptide sequences produced by mass spectrometry. We have used several complementary approaches to explore the global sporozoite proteome of C. parvum with available proteomic tools. To optimize the output of the MS data, a sequence similarity-based MS BLAST strategy was employed for bioinformatic analysis. Most significantly, almost all the constituents of glycolysis and several mitochondrion-related proteins were identified. In addition, many hypothetical Cryptosporidium proteins were validated by the identification of their constituent peptides. The MS BLAST approach was found to be useful during the study and could provide valuable information towards a complete understanding of the unique biology of Cryptosporidium.     

Effects of beta-glucan extracted from Saccharomyces cerevisiae on growth performance, and immunological and somatotropic responses of pigs challenged with Escherichia coli lipopolysaccharide

Three experiments were conducted to investigate the effects of beta-glucan supplementation on pig performance and immune function. In Exp. 1, 100 weaned pigs (8.65 +/- 0.42 kg of BW and 28 +/- 2 d of age) were used in a 35-d experiment to determine the effects of graded levels of beta-glucan. Pigs were randomly allotted to 1 of 5 treatments containing beta-glucan supplemented at 0, 25, 50, 100, or 200 ppm. Each treatment was replicated using 5 pens containing 4 pigs per pen. The ADG of pigs between d 14 to 28 and d 0 to 28 responded to dietary beta-glucan in a quadratic fashion (P < 0.05), whereas beta-glucan had no effect on ADFI and G:F in any period. In Exp. 2, 80 crossbred pigs (8.23 +/- 0.56 kg of BW and 28 +/- 2 d of age) were used in a 35-d experiment. Pigs were allotted to 1 of 2 dietary treatments (0 or 50 ppm of beta-glucan in the diet) using 10 pens with 4 pigs per pen. Pigs treated with beta-glucan had greater ADG in the 14- to 28-d (P = 0.05) and 0-to 28-d (P = 0.035) periods. The ADFI of pigs receiving beta-glucan was increased (P < 0.05) in the periods from 0 to 14, 0 to 28, and 28 to 35 d. The lymphocyte proliferation index in response to phytohemagglutinin (P = 0.051) and concanavalin A (P = 0.052) tended to decrease on d 14 in pigs supplemented with beta-glucan compared with pigs without supplementation. In Exp. 3, 24 barrows (8.89 +/- 0.20 kg of BW and 28 d of age) were used to investigate the immunological and somatotropic responses of pigs challenged with lipopolysaccharide (LPS). Experimental treatments were arranged in a 2 x 2 factorial, with the main effects of LPS challenge (saline vs. LPS) and dietary addition of beta-glucan (0 vs. 50 ppm). Pigs were raised individually in metabolic cages. Pigs were fed 0 or 50 ppm of beta-glucan for 28 d and then challenged with LPS (25 microg/kg of BW) or saline. After LPS injection, blood was obtained at 0, 1.5, 3, 4.5, 6, and 7.5 h to determine cytokine production and the somatotropic response. Dietary beta-glucan increased plasma interleukin-6 at 1.5, 3, and 4.5 h and tumor necrosis factor-alpha at 3 and 4.5 h and increased plasma interleukin-10 from 3 to 7.5 h after LPS challenge. The beta-glucan treatments had no effect on growth hormone. In conclusion, beta-glucan can selectively influence performance and partially offer benefits on somatotropic axis and immune function in weaned piglets challenged with LPS.     

Differential expression of peroxisome proliferator-activated receptors alpha and gamma gene in various chicken tissues

The peroxisome proliferator-activated receptors (PPARs) are the members of superfamily of nuclear hormone receptors. A great number of studies in rodent and human have shown that PPARs were involved in the lipids metabolism. The goal of the current study was to investigate the expression pattern of PPAR genes in various tissues of chicken. The tissue samples (heart, liver, spleen, lung, kidney, stomach, intestine, brain, breast muscle and adipose) were collected from six Arber Acres broilers (8 weeks old, male and female birds are half and half). Semi-quantitative RT-PCR and Northern blot were used to characterize the expression of PPAR-alpha and PPAR-gamma genes in the above tissues. By semi-quantitative RT-PCR, the results showed the expression level of PPAR-alpha gene was higher in brain, lung, kidney, heart and intestine, medium in stomach, liver and adipose than in spleen, and it did not express in breast muscle. The expression level of PPAR-gamma gene was higher in adipose, medium in brain and kidney than in spleen, heart, lung, stomach and intestine, but it did not express in liver and breast muscle. Northern blot results showed that PPAR-alpha gene expressed in heart, liver, kidney and stomach, and the intensity of hybridization signal was the stronger in liver and kidney than in other tissues, however, PPAR-gamma gene only expressed in adipose and kidney tissues. The results of this study showed the profile of PPAR gene expression in the chicken was similar to that in rodent, human and pig. However the expression profile of chicken also have its own specific trait, i.e. compared with mammals, PPAR-alpha gene can not be detected in skeletal muscle and PPAR-gamma gene can be stronger expressed in kidney tissues. This work will provide some basic data for the PPAR genes expression and lipids metabolism of birds.     

Outbreak of clinical listeriosis in sheep: evaluation from possible contamination routes from feed to raw produce and humans

We report the results of clinical and microbiological investigations on Listeria monocytogenes infections in a flock of 55 sheep and describe the implications for the safety of the raw milk and raw-milk cheeses produced in the on-farm dairy. The outbreak was caused by feeding grass silage, which was contaminated with 5 log10 CFU L. monocytogenes/g. Clinically, although having been fed from the same batch of silage, abortive (nine ewes), encephalitic (one ewe) and septicaemic (four ewes) forms of listeriosis were observed during the outbreak phase. As the starting point of feeding the contaminated silage was known we could calculate an incubation period of 18+/-2 and 26 days for the abortive and the encephalitic form of listeriosis, respectively. Pathologically, the septicaemic cases suffered from Listeria accumulation at comparable numbers in visceral organs but not in the brain. Only a single ewe developed central nervous symptoms and a rhomb-encephalitis was immunohistologically confirmed. In this case the infection proceeded from the nasal mucosa into the brain, with no infections of the liver, spleen and other visceral organs. Sampling of the cheese production chain, the farm environment and the persons living at the farm revealed the exposure of a farm-worker to an isolate genetically indistinguishable from the outbreak clone, obviously through the consumption of faecally contaminated bovine raw milk. The cheese under processing was free of Listeria because, as a result of intensive consultations, the farmer ensured a proper acidification of the cheese. The epidemiological findings suggest that food safety matters should be assessed in any case where infection of food-producing animals with potential human pathogens is observed.     

Dietary Supplementation with Microencapsulated Lutein Improves Yolk Color and Lutein Content in Fresh and Cooked Eggs of Laying Hens

This study was conducted to compare the efficacy of diet supplemented with non-microencapsulated lutein (NL) and microencapsulated lutein (ML) in laying hens. A total of 270 Hy-line Brown laying hens (54 weeks old) were allocated to three groups with six replicates of 15 hens and were adapted to a wheat-soybean meal basal diet for two weeks. Next, the control birds were fed the basal diet, and the test birds were fed the basal diet supplemented with 600 mg/kg NL (12 mg/kg available lutein) or 90.1 mg/kg ML (10 mg/kg available lutein) for 35 days. Supplementation of lutein did not affect the productive performance of laying hens, but improved (P<0.05) the yolk color and red/green value (a*), with eggs from the ML group displaying improved color and a* values from the 15^th day of the experimental period. The blue/yellow value (b*) for the yolk showed an increase (P<0.05) through both NL and ML supplements. The yolk color of fried and boiled eggs and a* value of the yolk in fried eggs were improved (P<0.05) only through ML supplemented diet. Both NL and ML supplements resulted in lower (P<0.05) lightness and higher (P<0.05) a* values of yolk in boiled eggs, as well as higher (P<0.05) b* values in fried and boiled eggs. Yolk lutein content in fresh, fried, and boiled eggs was increased (P<0.05) in NL and ML groups with the latter being higher. In conclusion, ML improved yolk pigmentation and lutein retention in laying hens better than NL.     

Illustration of the maternal animal model used for genetic evaluation of beef cattle

National cattle evaluation programs for weaning weight in most beef breed associations involve implementation of the maternal animal model to predict direct and maternal EPD. With this model, direct breeding values are predicted for all animals with records or pedigree ties to animals with records, or both. Even though maternal genetic value is expressed only in animals that become dams, these effects are transmitted by all parents and inherited from parents by all animals, leading to maternal breeding values being predicted for all animals as well. A small example data set was simulated involving 12 parents, 8 nonparents, and 13 animals with weaning weight records. The pedigree was developed to include paternal and maternal half-sib families, full-sibs, and some inbreeding, similar to field populations of beef cattle. Assembly of the mixed model equations and solutions for the maternal animal model are illustrated explicitly to assist animal breeding students in their understanding of the properties of the maternal animal model and to explicitly implement the model. Model parameters and moments, fixed contemporary group solutions, adjustment of breeding values for merit of mates, interpretation of maternal permanent environmental effect solutions, and alternatives for the assembly of the equations are shown. This example should lead to increased student and producer understanding of genetic improvement programs for weaning weight in beef cattle.     

Oesophagogastric parakeratosis in the growing pig: effects of the physical form of barley-based diets and added fibre

In four experiments a total of 288 individually fed pigs were given barley-based diets for about 100 days from about 20 kg liveweight. Fine grinding of barley increased the number and severity of oesophagogastric lesions. Pelleting a diet based on coarsely ground barley had a similar effect. Coarser grinding of the barley and substituting small proportions of oat husk, but not of bran, gave lower incidences and severities of lesions. The performance responses of the pigs differed and give a framework for deciding on the balance to be struck between optimal performance and the risk of lesion development.