A serosurvey of Borna disease virus infection in wild rats by a capture ELISA

For a serological diagnostic test for Borna disease (BD), we developed a capture ELISA with specificity and sensitivity based on detection of antibodies against BD virus (BDV) p40 protein. Using our capture ELISA system, the antibody response of rats inoculated intracerebrally with BDV at 4 weeks after birth showed a sharp increase from 1 to 4 weeks postinoculation (p.i.) and a steady level after 5 weeks p.i. To investigate prevalence of BDV infection among wild rats, we examined sera of Rattus norvegicus in Kami-iso town, Oshima district, Hokkaido, suggesting that rats in this area had not been infected by BDV.     

Efficacy of thiabendazole, mebendazole, levamisole and ivermectin against gullet worm, Gongylonema pulchrum: in vitro and in vivo studies

In vitro and in vivo studies were conducted to evaluate the effects of thiabendazole, mebendazole, levamisole and ivermectin against Gongylonema pulchrum. For in vitro assays, third-stage larvae (L3) incubated with the drugs were administered orally to mice and the ability of larvae to invade the gastric mucosa of the animals was examined. After incubation, only those larvae treated with high concentrations of levamisole (1 and 10 microg/ml) were tightly coiled with intestines exhibiting morphological abnormalities. Good dose-response data for the drugs tested was observed at the time of worm recovery from mice, with no worms recovered at the two highest concentrations of levamisole. In vivo efficacy of the drugs against adult worms was evaluated in six groups of three rabbits, each of which was infected with 30 L3 of G. pulchrum and treated with thiabendazole at 100 mg/kg for 3 days, mebendazole at 70 mg/kg for 3 days, levamisole as a single dose of 8 mg/kg, and subcutaneously injected ivermectin as a single dose of 0.2 mg/kg or vehicles of the drugs (control) at 4 months post-infection. Necropsy 14 days after treatment revealed that levamisole, mebendazole and ivermectin reduced worm burdens by 63.2%, 22.8% and 25.8%, respectively, with no reductions in worms observed with thiabendazole. The surviving worms were principally found in the esophagus with the remainder distributed among the buccal mucosa, the tongue, and/or pharyngeal mucosa in all groups. A number of morphologically abnormal eggs were observed within the uterus and ovijector in female worms recovered from the thiabendazole-treated group. These findings suggest that levamisole exhibits in vivo efficacy against G. pulchrum infection and that the larval invasion tests using mice could be used to screen for anthelmintic susceptibility of nematodes.     

Effects of supplemental vitamin E during the mature period on the reproduction performance of Taiwan Native Chicken cockerels

One-day-old Taiwan native male chicks were fed with maize-soybean rearing diets without supplemental vitamin E to 23 weeks of age. From 23 to 52 weeks of age, the cockerels (n = 90) were assigned at random to 5 dietary treatments and fed with maize-soybean diets supplemented with 0, 20, 40, 80 and 160 mg/kg of vitamin E (dl-alpha-tocopherol acetate). Pullets (225) of the same age were fed with standard diets throughout. They were artificially inseminated with one dose of 0.04 ml/bird intact and 5-fold diluted pooled semen at 31 to 43 weeks of age and at 49 weeks of age, respectively. The criteria evaluated included: semen quality, fertility and maximum and effective duration of fertility, blood characteristics, body and testes weight. Supplemental vitamin E did not affect cockerels' effective duration of fertility and percentage of fertility. However, when pullets were inseminated with diluted semen, supplementing 160 mg/kg vitamin E increased the maximum duration of fertility at 49 weeks of age. Cockerels receiving 40 to 160mg/kg supplements had higher sperm viability and motility after 39 weeks of age and those fed 80 mg/kg had higher sperm concentration at 39 weeks of age. Cockerels receiving supplements of more than 40 mg/kg vitamin E had higher body weight gain. Plasma cholesterol and testosterone were not affected by supplemental vitamin E. However, plasma luteinising hormone (LH) concentration was lower in cockerels fed 160 mg/kg. Lack of supplemental vitamin E over 39 weeks was associated with lower semen quality but did not reduce the proportion of fertile eggs laid by inseminated hens, perhaps because the insemination dose compensated for low sperm quality. We found that the maximum duration of fertility might be improved by supplementing 160 mg/kg vitamin E at 49 weeks of age.     

Effect of ensiling whole crop oat with lucerne in different ratios on fermentation quality,aerobic stability and in vitro digestibility on the Tibetan plateau

The objective of this study was to determine the effect of ensiling different ratios of whole crop oat to lucerne on fermentation quality, aerobic stability and in vitro digestibility of silage on the Tibetan plateau. Four experimental treatments were produced varying in the ratio of forages on a fresh matter (FM) basis: 1) 100% oat (control, dry matter (DM) content: 317 g/kg), 2) 90% oat + 10% lucerne (OL10, DM content: 316 g/kg), 3) 80% oat+ 20% lucerne (OL20, DM content: 317 g/kg) and 4) 70% oat+ 30% lucerne (OL30, DM content: 318 g/kg). All treatments were packed into laboratory‐scale silos and ensiled for 60 days and then subjected to an aerobic stability test for 15 days. Further, the four experimental treatments were incubated in vitro with buffered rumen fluid to study the nutrient digestibility. All silages were well preserved with low pH and NH₃‐N contents, and high lactic acid contents and V‐scores (evaluation of silage quality). Increasing the lucerne proportion increased (p < 0.05) crude protein (CP) content of silage, whereas neutral (NDF) and acid (ADF) detergent fibre contents were not affected. Under aerobic conditions, the control silage showed higher (p < 0.05) yeast counts (>10⁵ cfu/g FM) followed by OL10 silage, and OL10 silage improved aerobic stability for 74 h. OL20 and OL30 silages showed fewer (p < 0.05) yeasts (<10⁵ cfu/g FM) and markedly (p < 0.05) improved the aerobic stability (>360 h). After 48‐h incubation, OL30 silage increased (p < 0.05) in vitro dry matter digestibility (IVDMD) and neutral detergent fibre digestibility (IVNDFD) compared with the control silage. These results suggest that replacing oat with lucerne had no unfavourable effects on fermentation quality of silage, but improved CP content, aerobic stability IVDMD and IVNDFD. OL30 silage was the best among the three mixed silages.     

In vitro fertilization and cortical granule distribution of bovine oocytes having heterogeneous ooplasm with dark clusters.

In vitro maturation, fertilization and subsequent development of oocytes with homogeneous (category 1), or heterogeneous ooplasm (category 2) were investigated. No significant differences were observed in the nuclear maturation and total fertilization rates between the two categories. However, category 2 oocytes showed a higher normal fertilization rate due to their lower incidence of polyspermy as compared to category 1 oocytes. Electron microscopic study revealed that all category 2 oocytes had cortical granules lined up next to the plasma membrane, and that some category 1 oocytes still had small clusters of cortical granules after maturation. Although the proportion of cleaved zygotes was higher in category 2, the percentages of cleaved zygotes that developed to the blastocyst stage did not differ between the two categories. These results demonstrate that oocytes with heterogeneous ooplasm have a higher capacity for normal fertilization due to the reduction in polyspermy. This can be attributed to the normal distribution of cortical granules in category 2 oocytes after maturation.     

Feeding different dietary protein to energy ratios to Holstein heifers: effects on growth performance,blood metabolites and rumen fermentation parameters

Eighteen Chinese Holstein heifers average age 230 ± 14 days were allocated to 1 of 3 dietary crude protein (CP) to metabolizable energy (ME) ratios to examine the effects on growth performance, blood metabolites and rumen fermentation parameters with 90‐days experiment. Three different dietary CP:ME ratios were targeted based on the formulation of dietary CP contents of 10.85%, 12.78% and 14.63% on dry matter (DM) basis with similar ME contents (10.42 MJ/kg DM), which were categorized as low, medium and high dietary CP:ME ratios. The actual CP:ME ratios obtained in this study significantly increased from low to high CP:ME ratio groups with a value of 10.59, 11.83 and 13.38 g/MJ respectively. Elevated CP:ME ratios significantly increased CP intake (kg/day) and feed efficiency (FE) which was defined as dry matter intake as a proportion of average daily gain (ADG), whereas little difference was observed in body weight (kg), ADG (kg/day), DM intake (kg/day) and ME intake (MJ/day) among the three different CP:ME ratio groups. Increasing dietary CP to ME ratios significantly increased CP digestibility, whereas digestibility of DM and gross energy remained constant in the current experiment. Blood urea nitrogen and insulin‐like growth factor‐1 linearly increased with increasing dietary CP:ME ratios. There was significantly dietary treatment effect on rumen fermentation parameters including acetate, propionate, butyrate and total volatile fatty acids. Therefore, this study indicated that increasing dietary CP levels with similar energy content contributed to increased protein intake and its digestibility, as well as FE. Holstein heifers between 200 and 341 kg subjected to 13.38 dietary CP:ME ratio showed improved feed efficiency, nutrient digestibility, some blood metabolites and rumen fermentation characteristics for 0.90 kg/day rate of gain.     

Efficacy of combined chemotherapy against gastrointestinal nematodes and Fasciola hepatica in cattle

Direct blood smear examination (using 0.05 ml of whole blood) detected 168 (80.9%) of 204 microfilaremic canine blood samples as determined by the modified Knott test for microfilariae (mff) of Dirofilaria immitis (using 1 ml of whole blood). Direct smear examination detected all of 134 microfilaremias greater than 50 mff ml(-1), but only 31 of 70 (44.3%) microfilaremias having less than 50 mff ml(-1). In a separate retrospective query of a database of 963 dogs with necropsy-confirmed heartworm infections, 834 (86.6%) were positive by the DiroCHEK heartworm antigen test, and 504 (52.3%) were microfilaremic by the modified Knott test. Only 2 (0.4%) of the microfilaremic dogs were DiroCHEK negative and another 18 (3.6%) were very weak positives. Although these microfilaremic dogs were not tested by direct smear, only one of the two DiroCHEK-negative and six of 18 weakly DiroCHEK-positive dogs had microfilaremias so low that a direct smear may have given a false negative result. Significant adverse reactions to either diethylcarbamazine or the macrolide endectocides have not been reported for microfilaremias less than 500 mff ml(-1), thus substitution of the direct smear for a concentration test for mff, such as the modified Knott test or membrane filtration, does not appear to increase the risk of an unexpected adverse reaction to heartworm prophylactic drugs. Such a substitution results in only a very slight decrease (on the order of 0.1%) in the overall sensitivity of heartworm screening, provided a test for mff is run concurrently with an antigen test. If a test for mff is the only screening test used, then substitution of a direct smear for a concentration test may decrease the sensitivity of heartworm screening by nearly 20%, depending on the prevalence of low level microfilaremias in the population of dogs tested.     

Characterization of staphylococci associated with clinical and subclinical bovine mastitis

Attempts were made to identify 900 species of staphylococci or micrococci recovered from samples of bovine milk examined for mastitis pathogens. The presence and identity of haemolysins was recorded together with results of disc diffusion antibiotic sensitivity tests. The occurrence of clinical mastitis was also noted and somatic cell counts (SCC) were performed on milk samples which were normal in appearance. Eight hundred and thirty-one coagulase positive staphylococci were obtained, of which 810 were S. aureus and 21 were S. intermedius. Of 65 coagulase negative staphylococci the species of 19 could not be determined by the identification systems used. The remainder were identified as S. hyicus sub sp. hyicus (1), S. hyicus sub sp. chromogenes (19), S. haemolyticus (17), S. hominis (3), S. epidermidis (4), S. capitis (1) and either S. hominis or S. warneri (1). Four other isolates could not clearly be assigned to the genus Staphylococcus or Micrococcus and were designated irregular strains. No micrococci were identified. The presence of alpha, beta, or delta haemolysins occurring singly or in various combinations was identified in 98.3% of coagulase positive staphylococci and in 60% of coagulase negative staphylococci. Epsilon haemolysin was detected in 47.6% of the coagulase negative staphylococci and in 9.5% of S. intermedius. All staphylococci were sensitive to tetracycline (30 microg), novobiocin (1.6 microg), nafcillin (30 microg), methicillin (10 microg) and cephalothin (30 microg) and variable numbers of each species were sensitive to penicillin (2 iu) and streptomycin (10 microg). One non-identified species of coagulase negative staphylococcus was sensitive to erythromycin (0.4 microg) the remaining staphylococci were resistant. Each of the four irregular strains was sensitive to erythromycin and novobiocin. Clinical mastitis was associated with 30.6% of coagulase positive staphylococci, 15.3% of coagulase negative staphylococci, and two of the four irregular strains (50%). Subclinical mastitis as determined by SCC of 500 x 10(3) or greater was associated with 92.7% of coagulase positive and 37.5% of coagulase negative staphylococci.     

Impact of grassland contract policy on soil organic carbon losses from alpine grassland on the Qinghai–Tibetan Plateau

Carbon storage in the soils on the Qinghai–Tibetan Plateau plays a very important role in the global carbon budget. In the 1990s, a policy of contracting collective grasslands to smaller units was implemented, resulting in a change from the traditional collective grassland management to two new management patterns: a multi‐household management pattern (MMP: grassland shared by several households without enclosures) and a single‐household management pattern (SMP: grassland enclosed and used by only one household). In 2016, 50 MMP and 54 SMP winter pastures on the Qinghai–Tibetan Plateau were sampled to assess the differences in soil organic carbon (SOC) between the two management patterns. Results showed that average SOC was significantly greater under MMP than under SMP, with an estimated 0.41 Mg C/ha/yr lost due to SMP following the new grassland contract. Based on the government's grassland policy, four grassland utilization scenarios were developed for both summer and winter pastures. We found that if the grassland were managed under SMP, likely C losses ranged between 0.31 × 10⁷ and 6.15 × 10⁷ Mg C/yr across the Qinghai–Tibetan Plateau relative to MMP, which more closely resembles pre‐1990s grassland management. Previous estimates of C losses have only considered land use change (with cover change) and ignored the impacts driven by land management pattern changes (without cover change). The new data suggest that C losses from the Qinghai–Tibetan Plateau are greater than previously estimated, and therefore that the grassland contract policy should be reviewed and SMP households should be encouraged to reunite into the MMP. These findings have potential implications for land management strategies not only on the Qinghai–Tibetan Plateau but also other grazing regions globally where such practices may exist.