Evaluation of a competitive ELISA for antibody detection against avian influenza virus

Active serologic surveillance is necessary to control the spread of the avian influenza virus (AIV). In this study, we evaluated a commercially-available cELISA in terms of its ability to detect AIV antibodies in the sera of 3,358 animals from twelve species. cELISA detected antibodies against reference H1- through H15-subtype AIV strains without cross reactivity. Furthermore, the cELISA was able to detect antibodies produced following a challenge of the AIV H9N2 subtype in chickens, or following vaccination of the AIV H9 or H5 subtypes in chickens, ducks and geese. Next, we tested the sensitivity and specificity of the cELISA with sera from twelve different animal species, and compared these results with those obtained by the hemagglutination-inhibition (HI) test, the "gold standard" in AIV sera surveillance, a second commercially-available cELISA (IZS ELISA), or the agar gel precipitation (AGP) test. Compared with the HI test, the sensitivities and specificities of cELISA were 95% and 96% in chicken, 86% and 88% in duck, 97% and 100% in turkey, 100% and 87% in goose, and 91% and 97% in swine, respectively. The sensitivities and specificities of the cELISA in this study were higher than those of IZS ELISA for the duck, turkey, goose, and grey partridge sera samples. The results of AGP test against duck and turkey sera also showed significant correlation with the results of cELISA (R-value >0.9). In terms of flock sensitivity, the cELISA correlated better with the HI test than with commercially-available indirect ELISAs, with 100% flock sensitivity.     

The effect of probiotic BioPlus 2B® on growth performance,dry matter and nitrogen digestibility and slurry noxious gas emission in growing pigs

For this study, 2 experiments were conducted to evaluate the effects of dietary BioPlus 2B^® supplements on growing pigs. In experiment 1, 64 crossbred pigs (26.70 ± 0.50 kg) were subjected to a 35-day feeding trial (2 pigs/pen, 8 pens/treatment) in which the effects of the following four corn-soybean meal based dietary treatments were compared: 1) CON (basal diet); 2) B0.05 (basal diet + 0.05% BioPlus 2B^®); 3) B0.1 (basal diet + 0.1% BioPlus 2B^®) and 4) B0.2 (basal diet + 0.2% BioPlus 2B^®). During the experimental period, average daily gain (ADG) tended to increase in a linear fashion (P = 0.08) with increasing levels of dietary BioPlus 2B^®. In addition, the average daily feed intake (ADFI) was found to increase (P < 0.05) as the levels of BioPlus 2B^® increased. However, no linear or quadratic effects were observed in the gain:feed ratio (G:F) or in the coefficients of total tract apparent digestibility (CTTAD) when dry matter (DM) and nitrogen (N) were considered. In experiment 2, 16 crossbred barrows (25.60 ± 0.10 kg) were randomly allotted to 1 of the 4 dietary treatments used in experiment 1 (1 pig/pen, 4 pens/treatment). The noxious gas emissions and pH of slurry waste were then measured daily for 120 h. Ammonia (NH₃) emission from the slurry samples obtained from pigs in the BioPlus 2B^® groups was lower than that of slurry obtained from pigs in the CON treatment (P < 0.05), however, hydrogen sulfide (H₂S) and mercaptan emission were not affected by supplementation with BioPlus 2B^®. In addition, the pH value decreased with time, and the pH values of slurry obtained from pigs subjected to the B0.1 and B0.2 treatments were lower than those of samples obtained from other groups after 48 h (P < 0.05). Taken together, these results indicate that supplementation with BioPlus 2B^® can reduce slurry NH₃ emission, but not H₂S and mercaptan emission in growing pigs without impacting growth performance.     

Fatty acid composition and meat quality traits of organically reared Korean native black pigs

Current study investigated the effect of feeding systems (conventional vs organic rearing) on carcass characteristics, meat quality and fatty acid composition of Musculus longissimus dorsi of Korean native black barrows (KNP). Thirty pigs were reared under a conventional feeding system at indoor area of 1 m² per head, while another thirty pigs were fed an indoor area with organic saw dusts of 1 m² and an outdoor area with free ranges of 1 m² for each pig for organic system. Diet for the organic rearing was also provided according to the guideline for organic pork products. Warner–Bratzler shear force was lower and water holding capacity was higher for pork produced under the organic guidelines. However, pork produced by an organic system did not affect sensory traits compared to pork produced by a conventional one. In addition, longissimus muscle from organically reared pigs had significantly (P < 0.05) higher myoglobin content and consequently higher CIE a?-values compared with those for the conventionally-reared pigs (P < 0.05). The organic pork resulted in significantly (P < 0.05) higher polyunsaturated fatty acid (PUFA) and unsaturated fatty acid contents, as well as a higher n-3 PUFA than the conventional one (P < 0.05).     

Chitosan derivatives killed bacteria by disrupting the outer and inner membrane

Six kinds of water-soluble chitosan were prepared by grafting aminofunctionality onto chitosan at the C-6 position, and their antimicrobial activities were investigated against three Gram-negative and three Gram-positive bacteria. Among the derivatives, dimethylaminoethyl-chitosan (DMAEC) has the highest potential to suppress the growth of bacteria. To elucidate detailed antimicrobial modes of action against bacteria, cell integrity, outer membrane (OM), and inner membrane (IM) permeabilization assays were investigated. When treated with DMAEC, the release of 260 nm absorbing materials quickly increased for both Escherichia coli and Staphylococcus aureus, but the absorbance value was different due to the difference in cell structures. In OM and IM permeabilization assays, DMAEC rapidly increased 1-N-phenylnaphthylamine uptake and the release of cytoplasmic beta-galactosidase via an increase in the permeability of OM and IM. Moreover, DMAEC90 prepared from 90% deacetylated chitosan had more activity than DMAEC50 prepared from 50% deacetylated chitosan, and these results revealed that the antimicrobial action of water-soluble chitosans was dependent on the degree of deacetylation and the substituted group.     

Isoflavone profiles of red clovers and their distribution in different parts harvested at different growing stages

The isoflavone compositions and concentrations in the leaf, flower, petiole, and stem of 13 red clover cultivars were studied using high-performance liquid chromatography coupled with a diode array and a mass spectrometric detector with negative electrospray ionization. Different cultivars showed significantly different concentrations of individual and total isoflavones. The leaf contained the highest overall concentration, followed by the stem, petiole, and flower. Biochanin A and formononetin were the predominant isoflavones in all cultivars and all parts, along with eight other minor aglycones, daidzein, genistein, glycitein, irilone, orobol, pratensein, pseudobaptigenin, and prunetin, and four minor malonylglycosides, genistein-7-glucoside-6' '-malonate, orobol-7-glucoside-6' '-malonate, formononetin-7-glucoside-6' '-malonate, and biochanin A-7-glucoside-6' '-malonate. The isoflavone compositions and concentrations were also found to be different between red clover parts harvested at the early bud stage and the late flowering stage. Sample storage and handling prior to analysis were also found to be important. Samples in this study were kept at -5 degrees C for a few days before being freeze-dried and were found to contain mainly the aglycones of isoflavones. This may actually be an advantage in that "natural" and more bioactive isoflavones can be obtained without using chemical hydrolysis. Findings in this study therefore provide important information for developing isoflavone-rich red clovers and for optimizing harvest timing and choosing the right part of the red clover plant.     

Experimental infections with Lepeophtheirus salmonis (Krøyer) on threespine sticklebacks, Gasterosteus aculeatus L., and juvenile Pacific salmon, Oncorhynchus spp

Experimental infections with Lepeophtheirus salmonis (Krøyer) were established on threespine sticklebacks, Gasterosteus aculeatus L., juvenile pink, Oncorhynchus gorbuscha (Walbaum), and chum, Oncorhynchus keta (Walbaum), salmon. The prevalence and abundance of infections were initially higher on sticklebacks than on either salmon species. The initial prevalence and intensity of infections on chum salmon were higher than those on pink salmon, and declined on both species during louse development. The rate of parasite development to adult stages was similar on all species although development beyond the preadult stage was not observed on sticklebacks. These results confirm previous field observations on the occurrence and development of L. salmonis on threespine sticklebacks.     

Detection of transgenic and endogenous plant DNA in digesta and tissues of sheep and pigs fed Roundup Ready canola meal

The persistence of plant-derived recombinant DNA in sheep and pigs fed genetically modified (Roundup Ready) canola was assessed by PCR and Southern hybridization analysis of DNA extracted from digesta, gastrointestinal (GI) tract tissues, and visceral organs. Sheep (n = 11) and pigs (n = 36) were fed to slaughter on diets containing 6.5 or 15% Roundup Ready canola. Native plant DNA (high- and low-copy-number gene fragments) and the cp4 epsps transgene that encodes 5-enolpyruvyl shikimate-3-phosphate synthase were tracked in ruminal, abomasal, and large intestinal digesta and in tissue from the esophagus, rumen, abomasum, small and large intestine, liver, and kidney of sheep and in cecal content and tissue from the duodenum, cecum, liver, spleen, and kidney of pigs. High-copy chloroplast-specific DNA (a 520-bp fragment) was detected in all digesta samples, the majority (89-100%) of intestinal tissues, and at least one of each visceral organ sample (frequencies of 3-27%) from sheep and swine. Low-copy rubisco fragments (186- and 540-bp sequences from the small subunit) were present at slightly lower, variable frequencies in digesta (18-82%) and intestinal tissues (9-27% of ovine and 17-25% of porcine samples) and infrequently in visceral organs (1 of 88 ovine samples; 3 of 216 porcine samples). Each of the five cp4 epsps transgene fragments (179-527 bp) surveyed was present in at least 27% of ovine large intestinal content samples (maximum = 64%) and at least 33% of porcine cecal content samples (maximum = 75%). In sheep, transgene fragments were more common in intestinal digesta than in ruminal or abomasal content. Transgene fragments were detected in 0 (esophagus) to 3 (large intestine) GI tract tissues from the 11 sheep and in 0-10 of the duodenal and cecal tissues collected from 36 pigs. The feed-ingested recombinant DNA was not detected in visceral tissues (liver, kidney) of lambs or in the spleen from pigs. Of note, however, one liver and one kidney sample from the pigs (different animals) were positive for a 278-bp fragment of the transgenic cp4 epsps (denoted F3). Examination of genomic libraries from these tissues yielded no conclusive information regarding integration of the fragment into porcine DNA. This study confirms that feed-ingested DNA fragments (endogenous and transgenic) do survive to the terminal GI tract and that uptake into gut epithelial tissues does occur. A very low frequency of transmittance to visceral tissue was confirmed in pigs, but not in sheep. It is recognized that the low copy number of transgenes in GM feeds is a challenge to their detection in tissues, but there was no evidence to suggest that recombinant DNA would be processed in the gut in any manner different from endogenous feed-ingested genetic material.     

Prediction of human cases of West Nile virus by equine cases, Saskatchewan, Canada, 2003

In 2003, an outbreak of West Nile virus (WNV) occurred in Saskatchewan, Canada from July to September. One-hundred thirty-three horse cases and 947 human cases were recorded and data were analyzed retrospectively for evidence of clustering to determine if clinical infection in the horse population could be used to estimate human risk of infection with WNV. Kulldorff's scan statistic was used to identify spatial-temporal clusters in both the human and horse cases. In most areas, human clusters were not preceded by horse clusters. In one area, a significant cluster of horse cases preceded human cases by 1 week; however, 1 week does not provide sufficient time for human-health authorities to act and provide advance warning for the public.     

Potentiation of porcine circovirus 2-induced postweaning multisystemic wasting syndrome by porcine parvovirus is associated with excessive production of tumor necrosis factor-alpha

This study investigated the potentiation of porcine circovirus 2 (PCV2)-induced postweaning multisystemic wasting syndrome by porcine parvovirus (PPV) and found it was associated with excessive production of tumor necrosis factor-alpha (TNF-alpha). Colostrum-deprived conventional pigs were inoculated intranasally with PCV2 or PPV alone or in combination (PCV2 and PPV). In vitro assay of TNF-alpha, obtained from alveolar macrophages coinfected with PCV2 and PPV, showed a significant increase in TNF-alpha compared to single infection of macrophages with either PCV2 or PPV alone (P < 0.05). All pigs inoculated with PCV2 and PPV developed severe postweaning wasting syndrome, whereas clinical signs (e.g., weight loss) were present but perhaps less severe in either PCV2- or PPV-inoculated pigs. Compared to the pigs inoculated with PCV2 or PPV alone, pigs inoculated dually with PCV2 and PPV showed significantly (P < 0.05) increased levels of TNF-alpha. Levels of TNF-alpha in the sera were reversely correlated with the body weight in pigs experimentally infected with dual inoculation of PCV2 and PPV (r(s) = -0.92, P < 0.001). These data suggest that a potentiation of PPV in PCV2-induced PMWS is associated with the excessive production of TNF-alpha.     

Film preparation by melt- or compression-process using poly(vinyl chloride) powder swollen with dimethylformamide

A melt-process was used to prepare high molecular weight Poly(vinyl chloride) (PVC) films without the use of a conventional plasticizer and heat stabilizer. Rigid PVC powder was swollen with dimethylformamide containing 4∼10 vol% water to reduce its melting temperature. The swollen powder was pressed at a relatively low temperature of 75∼125 °C to form a film shape, and then washed and dried. The visible light transmittance, X-ray diffraction, density and the tensile properties of the resulting films were examined to estimate the success or failure of film formation. The films could be produced by not only the melt-process but also a compression-process using the rigid, highly swollen PVC powder. The resulting films had no voids, which are generally observed in PVC products formed by a solution process. The minimum temperature for these processes decreased with decreasing water content in the mixture: The minimum temperatures according to the water content in the mixture to produce faultless films through the melt-process were 4 %–105 °C, 6 %–115 °C, 8 and 10 %–125 °C, while those through the compression process were 4 %–95 °C, 6 and 8 %–105 °C, 10 %–115 °C.