Gene and haplotype polymorphisms of the Prion gene (PRNP) in Japanese Brown, Japanese native and Holstein cattle

Polymorphisms in the prion protein gene ( PRNP ) are known to be associated with transmissible spongiform encephalopathies in human, sheep and goats. There is tentative association between PRNP promoter polymorphism and bovine spongiform encephalopathy (BSE) susceptibility in cattle. In this study, we genotyped for six bovine PRNP polymorphic sites including a 23-bp indel in the promoter, a 12-bp indel in the intron 1, two nonsynonymous single nucleotide polymorphisms (SNPs), octapeptide repeats in the coding region and a 14-bp indel in the 3'-untranslated region in 178 animals representing Japanese Brown, Kuchinoshima feral, Mishima, Japanese Shorthorn and Holstein. In 64 Japanese Brown cattle, three indel sites were polymorphic. All of the six sites were monomorphic in Kuchinoshima. The 23-bp and 12-bp indel sites were polymorphic in Mishima cattle. The 23-bp and 14-bp indel sites were polymorphic in Japanese Shorthorn cattle. Both SNP sites were monomorphic in all cattle examined in this study. At the 23-bp indel site, the genotype frequencies of Japanese Brown and Holstein breeds were similar to that of BSE affected cattle. We estimated 12 different haplotypes from these genotypic data. A '23-12-K6S14+' haplotype was the major haplotype in all populations, whose frequencies ranged from 0.50 to 1.00.     

Evaluation of maturity of poultry manure compost by phospholipid fatty acids analysis

To study the influence of the physical properties of compost feedstock on some characteristics associated with maturity, two types of compost were made from poultry manure, rice husk, and rice bran. The bulk density of one type (PMC) was always higher than that of another type (NMC) during composting. In the case of PMC, the change in temperature, decrease in NH₄⁺, appearance of NO₃^–, and increase in germination indices (GI) with Japanese Komatsuna (Brassica campestris cv. Osome) were all more delayed than in NMC. As the composting process progressed, the proportion of branched (iso-, anteiso-, 10Me-) and saturated phospholipid fatty acids (PLFA) [BRANCHED FAMES (fatty acid methyl esters), biomarkers for gram-positive bacteria] gradually increased, then reached plateau. The high proportion of BRANCHED FAMES was maintained over a long storage period. The straight hydroxyl and saturated PLFAs (SOH-FAMES) initially increased, then disappeared with the progress of composting. The increase in BRANCHED FAMES and the decrease in SOH-FAMES were more delayed in PMC than NMC. The day on which the proportion of BRANCHED FAMES reached plateau and the proportion of SOH-FAMES dipped below 2 mol% coincided with the maturity stage based on the changes of physicochemical characteristics and GI. The composition of BRANCHED FAMES showed highly positive and negative correlation with GI and NH₄⁺, respectively. In the case of SOH-FAMES, inverse correlations were observed. This indicates that the proportion of BRANCHED FAMES and/or SOH-FAMES can be used as a tool for evaluating the maturity of poultry manure compost.     

Molecular and immunochemical characteristics of monoclonal and recombinant antibodies selective for the triazine herbicide simetryn and application to environmental analysis

A monoclonal antibody (mab) selective for the thiomethyl-s-triazine herbicide simetryn was obtained and characterized in enzyme-linked immunosorbent assay (ELISA). An IC(50) value for simetryn was 8.5 ng/mL, and the detection range extended from 1.1 to 70 ng/mL in ELISA. The cDNAs encoding variable heavy chain (VH) and variable light chain (VL) of the mab were cloned to produce various recombinant antibodies. Single-chain variable fragment (scFv) antibodies derived from the mab were characterized in ELISA and showed similar reactivities and specificities to the parent mab. A urea denaturation test revealed that the scFv antibodies bound to simetryn were more stable than those in the absence of antigen. A sandwich ELISA based on VH and VL fragments of the mab was successfully developed and showed similar sensitivity to those based on the mab and scFv antibodies in ELISA. In the recovery experiments using spiked environmental samples, the results obtained in ELISA based on the mab were favorably correlated with those by HPLC.     

Effects of early high nutrition related to metabolic imprinting events on growth,carcass characteristics,and meat quality of grass-fed Wagyu (Japanese Black cattle)

The study was conducted to clarify how early high plane of nutrition related to metabolic imprinting affected growth, carcass characteristics, and meat quality of grass-fed Wagyu (Japanese Black cattle). Wagyu steers were allocated randomly into 2 dietary groups: (1) steers fed milk replacer (crude protein 26.0%, crude fat 25.5%; maximum intake 0.6 kg/d) until 3 mo of age and then fed roughage (orchard grass hay) ad libitum from 4 to 10 mo of age (roughage group, RG; n = 11); (2) steers fed milk replacer (maximum intake of 1.8 kg/d) until 3 mo of age and then fed a high-concentrate diet from 4 to 10 mo of age (early high nutrition, EHN; n = 12). After 11 mo of age, all steers were fed roughage ad libitum until 31 mo of age and then slaughtered. Growth performance, carcass traits, longissimus muscle (LM) meat quality and intramuscular fat (IMF) content, plasma insulin-like growth factor I (IGF-I) concentration, and bone mineral density were measured. Body weight was greater in EHN steers (571 kg) than RG steers (520 kg; P < 0.01). Plasma IGF-I levels were higher in EHN steers than in RG steers at 3, 10, and 14 mo of age (P < 0.01, P < 0.005, P < 0.001, respectively); however, plasma IGF-I levels were lower in EHN steers compared with RG steers at 30 mo of age (P < 0.01). The total weight of the muscles and bones of the left half of the carcass was not different between the 2 groups (P = 0.065). Five of the 19 muscles investigated (semimembranosus, P = 0.036; infraspinatus, P = 0.024; supraspinatus, P = 0.0019; serratus ventralis cervicis, P = 0.032; serratus ventralis thoracis, P = 0.027) were heavier in EHN steers. Total fat weight in the left half of the carcass was 30% greater (P = 0.025) in HNE carcasses. Subcutaneous and perirenal fat weights were 53% and 84% greater (P = 0.008, P = 0.002, respectively) in EHN carcasses. The LM IMF content was greater in EHN loins (13.2%) compared with RG loins (9.4%) at 31 mo of age (P = 0.038); however, no differences were found for shear force, tenderness, and cook loss. These results suggested early high-nutrition affected the growth and meat quality of livestock.     

Molecular epidemiological survey of Theileria orientalis in Thua Thien Hue Province, Vietnam

Theileria orientalis is a benign bovine protozoan parasite that occasionally causes serious economic loss in the livestock industry. We report the findings of a molecular epidemiological survey of T. orientalis in 94 Vietnamese yellow cattle, 43 water buffaloes, 21 sheep, 21 goats and 85 blood-sucking ticks of cattle in the Thua Thien Hue province of Vietnam. The major piroplasm surface protein (MPSP) gene of T. orientalis was detected using polymerase chain reaction from 13 cattle (13.8%), 11 water buffaloes (25.6%), 1 sheep (4.8%) and 9 ticks (10.6%). Phylogenetic analysis using MPSP gene sequences showed the presence of seven genotypes, four previously categorized genotypes (Types 1, 3, 5 and 7) and three new genotypes (Types N-1, N-2 and N-3).     

Shedding of Neospora caninum oocysts by dogs fed different tissues from naturally infected cattle

Neospora caninum is one of the most important causes of abortion in dairy cattle worldwide. The distribution of N. caninum in tissues of adult cattle is unknown and the parasite has not been demonstrated histologically in tissues of cows. In the present study the distribution of N. caninum in different tissues of adult cattle was evaluated by bioassays in dogs. Seventeen dogs (2-3 month-old) were fed different tissues of 4 naturally exposed adult cattle (indirect fluorescent antibody test N. caninum titer ≥ 400): 5 were fed with masseter; 5 with heart, 3 with liver, 4 with brain, and 3 pups were used as non-infected control. Two dogs fed masseter, 2 fed heart, 1 fed liver, and 3 fed brain shed oocysts, and all dogs presented no seroconvertion to N. caninum during the observation period of 4 weeks. The oocysts were confirmed as N. caninum based on the detection of N. caninum-specific DNA by PCR and sequencing. The results indicate that dogs can be infected by N. caninum with different tissues of infected cattle.     

The Effect of Light on the Accumulation of Lipids in Rice Seeds

To investigate the effects of light on the accumulation of assimilate in rice seeds, ears were allowed to grow in darkness 10 d after their formation. The lipid content of rice bran on the 40th day increased by cultivation of ears in darkness whereas the dry weight and starch and sucrose contents of the rice seeds decreased. In the light, the triacylglyceride (TG) content of rice seeds was lowest on the 20th day but then increased continuously for the next 20 d, while the diacylglyceride (DG) content reached a maximum value on the 20th day. In darkness, in contrast, rice seeds showed a higher TG content, with a lower DG content and lower levels of other lipids on the 20th day compared with seeds exposed to light. These results suggest that the biosynthesis of starch and lipids in rice seeds is regulated by light.     

Topographic variation of soil nitrogen mineralization and microbial biomass in Japanese cedar (Cryptomeria japonica D. Don) stands at Myougodani watershed,Toyama, Japan

The spatial variability of nitrogen (N) mineralization, nitrification, and microbial biomass was investigated using surface soils from various topographic positions at a relatively small watershed with Japanese cedar (Crgptomeria japonica D. Don) plantations. The watershed topography was characterized using a topographic index derived from GIS analysis. The topographic index reasonably reflected the spatial variability of the soil water conditions, total soil carbon (C) and N contents, and exchangeable base concentrations. However, this index was not significantly correlated with the spatial variability of net N mineralization and microbial biomass. Topographic index and soil properties (total soil C and N contents, C / N ratio, exchangeable base concentrations, and clay content) were subjected to principal component analysis to eliminate multiple-collinearity among the variables, and express the variables as new orthogonal variables. Principal component analysis showed that the soil properties could be divided into two groups: PC1 (soil nutrient pools) and PC2 (soil clay content). The topographic index was closely correlated with PC1 and not significantly correlated with PC2. Regression of PC scores on net N mineralization and microbial biomass indicated the relatively high contribution of PC2 to the variability in N mineralization and microbial biomass. This result suggested that not only topographic factors but also the clay content exerted an important influence on the spatial pattern of N mineralization and microbial biomass within a watershed with single species forests.     

Effects of copper on survival and growth of larval false clown anemonefish <Emphasis Type="Italic">Amphiprion ocellaris</Emphasis>

ABSTRACT:   Larval false clown anemonefish of about 12 h after hatching were fed newly-hatched brine shrimp nauplii in natural (control) and copper-added seawater (40–640 µg–Cu/L) at 26.5 ± 0.5°C for 14 days. Survival rate of the fish increased with increasing copper concentrations up to 160 µg–Cu/L. The rates at 80 and 160 µg–Cu/L were 65 and 80%, respectively, and were significantly higher than that of the control (30%). A positive effect of copper addition on the survival rate was also observed in the other rearing experiment with the fish from seven different spawning chances. Growth rate of the survival fish was not affected by copper concentrations. The copper concentration of the survived fish increased with increasing copper concentrations in the rearing water.     

Effect of Feeding Frequency, Water Temperature, and Stocking Density on the Growth of Tiger Puffer, Takifugu rubripes

Effects of daily feeding frequency, water temperature, and stocking density on the growth of tiger puffer, Takifugu rubripes, fry were examined to develop effective techniques to produce tiger puffer in a closed recirculation system. Fish of 4, 14, and 180 g in initial body weight were fed commercial pellet diets once to five times a day to apparent satiation each by hand for 8 or 12 wk at 20 C. Daily feeding frequency did not affect the growth of 14‐ and 180‐g‐size fish. However, the daily feed consumption and weight gain of the 4‐g‐size fish fed three and five times daily were significantly higher than those of fish fed once daily (P < 0.05). Fish of 4 and 50 g in initial body weight were reared with the pellet diet at 15–30 C for 8 wk. The weight gain of fish increased with increasing water temperature up to 25 C and decreased drastically at 30 C for both sizes. Similar trends were observed for feed efficiency, although 4‐g fish had highest efficiency at 20 C. Effects of stocking density on growth were examined with fish of 8, 13, and 100 g in initial body weight. Fish were reared with the pellet diet for 8 or 16 wk at 20 C. Fish were placed in floating net cages in the culture tank, and the stocking density was determined based on the total weight of fish and volume of the net cage. Fish of 8 g in body weight grew up to 35–36 g during the 8‐wk rearing period independent of the stocking density of 8, 15, and 31 kg/m³ at the end of rearing. Final biomass per cage reached 32, 60, and 115 kg/m³ for 13‐g‐size fish, and 10, 18, and 35 kg/m³ for 100‐g‐size fish, and the growth of the fish tended to decrease with increasing stocking density for both sizes.