Serodiagnosis of Neospora caninum infection in cattle by enzyme-linked immunosorbent assay with recombinant truncated NcSAG1

Neospora caninum is a veterinary medically important pathogen capable of causing abortion in cattle and neuromuscular paralysis in dogs. The surface antigen 1 of N. caninum (NcSAG1) is an important candidate for the development of a diagnostic reagent for neosporosis. In order to establish an effective diagnostic method, the gene encoding truncated NcSAG1 (NcSAG1t) lacking a signal peptide and C-terminal hydrophobic regions was cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). The purified GST-NcSAG1t was tested in an enzyme-linked immunosorbent assay (ELISA) for the detection of N. caninum antibodies in cattle. The ELISA with GST-NcSAG1t clearly differentiated between immunofluorescent antibody test (IFAT)-positive and -negative sera from cattle. In addition, the ELISA detected no cross-reactivity with sera from mice experimentally infected with the closely related parasite Toxoplasma gondii. Field serum samples collected from cattle in Brazil were examined for the diagnosis of neosporosis by using the ELISA. Of the 197 samples analyzed, 66 (33.5%) samples were positive for antibodies to N. caninum. Of the 66 ELISA-positive samples, 60 (90%) samples were confirmed as positive by Western blot analysis with whole parasite antigens. These results suggest that the recombinant NcSAG1t could be a reliable reagent for use as an antigen in ELISA for the serodiagnosis of N. caninum infection in cattle.     

Distribution of Arcobacter species among livestock in Japan

A survey was conducted to examine the distribution of Arcobacter species among livestock in Japan. During May 1999 and May 2000, fecal samples from cattle (n=332) and swine (n=250), chicken cloacal swabs (n=234), and vaginal swabs of cattle (n=61) and swine (n=15) were submitted for the isolation of Arcobacter species. Arcobacter species were isolated from 3.6 and 10.0% of the cattle and swine fecal samples, respectively, along with 14.5% of chicken cloacal swabs. No significant seasonal differences were observed. Species-specific polymerase chain reaction assay showed that A. butzleri was the most prevalent species (83.3, 60.0 and 47.1% of the cattle, swine and chicken isolates, respectively), followed by A. cryaerophilus 1B (16.7, 36.0 and 55.9% of the cattle, swine and chicken isolates, respectively). Of the samples from vaginal swabs, 8.1 and 13.3% were positive for Arcobacter in cattle and swine, respectively. This is the first report demonstrating the distribution of Arcobacter species among livestock in Japan.     

Seroprevalence of Neospora caninum infection in dairy and beef cattle in Paraguay

Seroprevalence of Neospora caninum in 879 beef and dairy cattle in different locations of Paraguay was determined by an ELISA. In the survey, 262 (29.8%) cattle were positive to N. caninum, and animals with anti-Neospora antibody titre were observed in all the locations tested in the country. Serum samples taken from a herd that exhibited persistent abortion had the highest percentage of animals being positive to the parasite (17/30, 56.7%). In the same herd, abortion was significantly more likely in animals with high anti-Neospora antibody titre. Immunoblot analysis demonstrated that the banding pattern from positive Paraguayan cattle was similar to that seen with the positive control sample. In conclusion, N. caninum infection is present among Paraguayan beef and dairy cattle, and it may be an important cause of bovine abortion in Paraguay.     

Methane emission from stored dairy manure slurry and slurry after digestion by methane digester

To examine the effect of storage temperature on the emission of methane (CH₄) and carbon dioxide (CO₂) and on recovery of nutrients from raw dairy manure slurry (RS) and slurry digested in a methane digester (DS), both slurries were stored in closed 100 L steel tanks under psychrophilic conditions (5, 10, 15 and 20°C) for a 150 day period. As the storage temperature increased, total methane emission increased in both types of slurry. The amount of methane emitted per unit of volatile solids of the RS and DS was 0.19 L/g and 0.10 L/g, respectively. The respective carbon dioxide emissions were 0.20 L/g for RS and 0.12 L/g for DS at 20°C of storage temperature. At temperatures greater than 15°C, the methane concentration in the emitted gas remained more than 40% of the total gas. During the experimental period, in excess of 90% of the total Kjeldahl nitrogen in these slurries was recovered (91.4–93.7% for RS and 93.7–98.4% for DS) after storage, and ammonium nitrogen was recovered in excess of 100% (100.1–143.2% for RS and 106.7–143.2% for DS storage tanks) because of the mineralization of organic nitrogen in the influent. These results indicate that manure slurry characteristics and storage temperature have significant impacts on methane emission. It can be concluded that on typical farms located in northern Japan, methane emission from manure storage tanks during late fall, winter and early spring may be negligible, because of manure temperatures less than 10°C. During late spring, summer and early fall, methane emissions can be substantially reduced by using underground storage to maintain lower manure temperatures.     

Molecular biology and biotechnology for reduction of Fusarium mycotoxin contamination

Fusarium head blight (FHB) is a devastating disease of important cereal crops resulting in significant yield loss and mycotoxin contamination. Persistent outbreaks of FHB in Europe and North America have led to various efforts to understand the mechanisms of resistance to this disease and mycotoxin biosynthesis. In this minireview, we summarize basic and applied studies conducted in our laboratories into reducing mycotoxin contamination in FHB.     

The preventive effect of Bacillus subtilus strain DB9011 against experimental infection with enterotoxcemic Escherichia coli in weaning piglets

Porcine edema disease (ED) is caused by Shiga toxin 2e‐producing Escherichia coli (STEC). Post‐weaned piglets often suffer from ED as a result of intestinal infection with STEC, which causes impaired growth performance and high mortality. Antimicrobial therapy is a curative treatment for piglets infected with STEC, but the emergence of antimicrobial‐resistant STEC has become a serious problem for Japanese pig farmers. Therefore, an alternative strategy other than antimicrobial therapy is needed for the prevention or treatment of ED. In this study, we evaluated the effect of oral administration of Bacillus subtilis DB9011 (DB9011) to prevent the experimental infection of STEC in weaning piglets. Eight 21‐day‐old piglets were divided into two groups: STEC challenge with the basal diet, and STEC challenge with DB9011 supplemented diet. The challenge was carried out when the animals were 25, 26 and 27 days old using STEC contained in capsules resistant against gastric digestion. All pigs were euthanized at 36 days of age. DB9011 improved the symptoms of ED and decreased the number of STEC in the ileal digesta and feces. Accordingly, oral administration of DB9011 in weaned piglets prevents ED through the suppression of the growth of STEC in the ileum.     

Site-level model–data synthesis of terrestrial carbon fluxes in the CarboEastAsia eddy-covariance observation network: toward future modeling efforts

Based on the model–data comparison at the eddy-covariance observation sites from CarboEastAsia datasets, we report the current status of the terrestrial carbon cycle modeling in monsoon Asia. In order to assess the modeling performance and discuss future requirements for both modeling and observation efforts in Asia, we ran eight terrestrial biosphere models at 24 sites from 1901 to 2010. By analyzing the modeled carbon fluxes against the CarboEastAsia datasets, the strengths and weaknesses of terrestrial biosphere modeling over Asia were evaluated. In terms of pattern and magnitude, the carbon fluxes (i.e., gross primary productivity, ecosystem respiration, and net ecosystem exchange) at the temperate and boreal forest sites were simulated best, whereas the simulation results from the tropical forest, cropland, and disturbed sites were poor. The multi-model ensemble mean values showed lower root mean square errors and higher correlations, suggesting that composition of multiple terrestrial biosphere models would be preferable for terrestrial carbon budget assessments in Asia. These results indicate that the current model-based estimation of terrestrial carbon budget has large uncertainties, and future research should further refine the models to permit re-evaluation of the terrestrial carbon budget.     

Variation in lysine,threonine, and tryptophan availability in meat and bone meal as estimated by the slope‐ratio growth assay technique in growing rats

Meat and bone meal (MBM) is an important protein source used in animal feeds. However, as the composition and availability of amino acids (AAs) in MBM fluctuate markedly, it is important to verify the magnitude of these parameters in different MBMs. In this study, the AA compositions of 19 MBMs were analyzed to confirm variations in lysine (Lys), threonine (Thr), and tryptophan (Trp), then which were compared with those of soybean meal (SBM) and fish meal (FM). Instability in Lys, Thr, and Trp availabilities in six MBMs were also considered after estimation using the slope‐ratio growth assay technique in rats. Variations in AA composition were evaluated using the coefficient of variance (CV: Standard deviation/Mean). CVs for Lys, Thr, and Trp content were 9.40, 11.83, and 18.12 in MBM, 2.71, 2.48, and 3.19 in SBM, and 10.09, 10.44, and 13.47 in FM. Furthermore, means and SDs for Lys, Thr, and Trp availabilities in MBM were 53.3 ± 10.4% (CV: 19.5), 65.9 ± 17.6% (CV: 26.6), and 83.2 ± 11.2% (CV: 13.5), respectively. These results provide the first evidence that variations in MBM AA compositions were 3.5 to 5.7 times higher than those in SBM, but similar to FM, and that the large variation in availability substantially existed.     

Estimation of genetic parameters of defective appearances in Japanese Black heifer calves in Kagoshima

Defective appearances, including white spotting (WS), tongue defect (TD), and nipple defect (ND), in Japanese Black cattle potentially lead to economic losses to farmers in Japan. We estimated genetic parameters of defective appearances using 553,433 records of Japanese Black heifer calves housed in the Kagoshima Prefecture. Variance and covariance were estimated using the Gibbs sampling algorithm. The estimated heritability ranged from 0.29 for TD to 0.76 for WS. Percent breeding value (%BV) estimates indicated high variation in WS and ND among sires, reflecting higher heritability. Furthermore, there was a positive linear relationship between the %BV estimate of a sire and the mean incidence rate of each defect in his female offsprings. TD was positively associated with other defects. Therefore, genetic factors strongly affect the incidence of defective appearances in Japanese Black cattle.     

Examination of milk microbiota,fecal microbiota,and blood metabolites of Jersey cows in cool and hot seasons

Microbiota of individual cow milk, bulk tank milk, and feces of Jersey cows were examined. Samples were collected from two farms (F1 and F2) in cool (November, Nov) and hot (July, Jul) seasons. Milk yield and milk composition were similar between the two farms and between the two seasons. Prevalent taxa of the fecal microbiota, i.e. Ruminococcaceae, Bacteroidaceae, Lachnospiraceae, Rikenellaceae, and Clostridiaceae, were unaffected by the farm and season. Relative abundance of milk microbiota for Pseudomonadaceae, Enterobacteriaceae, and Streptococcaceae (F1 > F2) and Lactobacillaceae, Bifidobacteriaceae, and Cellulomonadaceae (F1 < F2) were different between the two farms, and those for Staphylococcaceae, Bacillaceae, Ruminococcaceae, and Veillonellaceae (Nov < Jul) and Methylobacteriaceae and Moraxellaceae (Nov > Jul) were different between the two seasons. The microbiota of bulk tank milk was numerically different from that of individual cow milk. Principal coordinate analysis indicated that the milk microbiota was unrelated to the fecal microbiota. The finding that relative abundance of Pseudomonadaceae and Moraxellaceae appeared greater than those reported for Holstein milk suggested that higher protein and fat content may result in a greater abundance of proteolytic and lipolytic taxa in Jersey cow milk.