Relationship between fat accumulation in the liver and energy intake, milk fat yield and blood metabolites in dairy cows

In the present study, 42 multiparous Holstein cows were used to investigate the relationship between fat accumulation in the liver and dry matter intake, milk yield and blood metabolites. Based on the percentage of fat in the liver cell at 2 weeks post‐parturition, the cows were classified into three groups. These groups were: (i) less than 10% of fat (normal group, n = 29); (ii) 10–20% of fat (mild group, n = 6); and (iii) more than 20% of fat (moderate group, n = 7). The bodyweight of the moderate group was high (771 kg) before calving. The sufficiency rates of total digestible nutrients (TDN) were remarkably decreased (approximately 65%) in early lactation. The milk fat yield and milk fat composition of the moderate group were higher (P < 0.05) than the other groups at 1 and 2 weeks post‐parturition. It was suggested that non‐esterified fatty acids (NEFA) mobilized from adipose tissues was directly used by the mammary gland for synthesis of milk fat. The percentage of bromsulfalein (BSP) retention of the moderate group was high (21.1%) at 30 min, and it showed that the BSP clearance function was significantly decreased. The concentrations of NEFA, β‐hydroxybutyric acid and glucose were appropriate indicators of energy status; however, aspartate aminotransferase, γ‐glutamyl transpeptidase and total bilirubin were not sensitive indicators of a moderate fatty liver. Thus, high‐yielding cows that calve in an overweight condition are more likely to develop excessive fat accumulation in the liver because of great mobilization from adipose tissues post‐parturition. In cows with a moderately fatty liver, a decrease in TDN sufficiency rates, an increase of milk fat yield and a reduction of liver function were observed in early lactation. The increase of serum NEFA and milk fat composition resulting from mobilization of adipose tissues helped to diagnose moderate fatty liver.     

The absence of Rhodococcus equi in Mongolian horses

In native Mongolian horses, the incidence and distribution of Rhodococcus equi are poorly understood. One hundred and fourteen equine fecal samples and 71 soil samples were collected from the camp sites of 26 nomadic families located in three areas less than 100 km from Ulaanbaatar, Mongolia. Five fecal samples were also collected from foals of Przewalski's Horses introduced into the Hustai National Park, Mongolia. No R. equi was isolated from the Mongolian horses or the soil samples. However, three colonies of R. equi were isolated from two fecal samples collected from foals of Przewalski's Horses. These isolates were avirulent, with neither 15- to 17-kDa antigens (VapA) nor a 20-kDa antigen (VapB) genes being detected. We concluded that native Mongolian horses and their environment appear free from contamination with R. equi.     

Urinary excretion of purine derivatives and plasma allantoin level in sheep and goats during fasting

The relationship between blood plasma level and urinary excretion of allantoin (AN) was examined in sheep and goats during fasting to investigate the possible use of purine derivatives (PD) in urine and/or plasma for estimating the microbial protein production in the rumen, and the further digestion in the lower guts of ruminants. Urinary AN excretion decreased markedly during fasting (0.13 mmol/kgW^0.75 per day), although urinary levels of other PD, hypoxanthine + xanthine and uric acid did not differ irrespective of the feeding condition, that is, feeding, fasting and refeeding in both species. The AN concentration in blood plasma also decreased drastically in the starvation period, and was suddenly increased on refeeding in sheep and goats, and these phenomena were very similar to those of urinary AN excretion. Therefore, there was a high positive correlation between plasma AN level and urinary AN excretion, and the coefficient of correlation was statistically significant (P < 0.01). These results clearly indicate that changes in urinary AN reflect change in plasma AN, which is induced by the catabolism of purine base in the body.     

A case study of infection of rice roots by deleterious fungi and minor pathogenic<Emphasis Type="Italic"> Pythium</Emphasis> species in a paddy field

Roots of rice plants grown in paddy fields in a transplant culture system were collected seven times between 8 and 22 weeks after transplanting, for two crop seasons. Rice seedlings grown in a potting medium amended with the collected roots were significantly shorter than those grown in the same medium either without the addition of the roots or amended with pasteurized, collected roots indicating that seedlings were inhibited by heat-labile microorganisms on the collected rice roots. Ninety-five and 172 pure cultures of Pythium spp. and fungi, respectively, were isolated from the rice roots collected 5 or 7.5 weeks after transplanting in the fields. Among these microorganisms, Pythium aristosporum inhibited seedling growth in greenhouse experiments, and Acremonium sp., Alternaria sp., Epicoccum nigrum, Fusarium sp., Massarina sp., Penicillium spp., Rhinocladiella sp., Stemphylium sp., Trichocladium sp., and several unidentified fungi inhibited seminal root growth in in vitro experiments. These microorganisms might be involved in the inhibition of seedling growth in soils amended with the rice roots collected from the paddy field. Thus, roots of rice plants at the middle stage of growth transplanted into paddy fields can harbor pathogenic or deleterious fungi or Pythium sp(p). The effects of these microorganisms on rice growth in paddy fields are discussed.     

Feed intake, lactation performance, blood metabolites and fertility in early lactation dairy cows grazing a timothy pasture

The present study was conducted to investigate feed intake, milk yield, milk composition, blood metabolites and fertility in early lactation dairy cows grazing a timothy pasture. Fourteen multiparous Holstein cows that calved between 20 May and 19 July were used over a 3‐year period. The stocking rate was 3.6–4.3 cow/ha. Concentrates were fed separately at 9.5–11.5 kg/day per cow (dry matter basis) from 1 to 13 weeks postparturition. Herbage intake was estimated using chromium oxide as an indigestible marker. The mean contents of crude protein, total digestible nutrients and neutral detergent fiber of pasture during the 3‐year study period were 22.3%, 71.8% and 51.7%, and those of total diet were 18.9%, 77.3% and 40.3%, respectively. The mean herbage dry matter intake was 13.0 kg/day from 2 to 13 weeks postparturition during the study, total dry matter intake was 23.7 kg/day, the total digestible nutrients sufficiency rate was 105%, milk yield was 39.7 kg/day, and milk fat percentage was 3.30%. The decrease in bodyweight postparturition was slight. Urea nitrogen concentrations in serum were below 18.3 mg/dL. The mean days to first estrus and days open were 36 and 104 days, respectively. These results indicate that energy deficiency, decrease in bodyweight and fertility in early lactation barely occur when high producing dairy cows are fed enough grazing grass and suitable concentrates.     

Perfusion analysis of periportal and centrilobular metabolism of paraoxon in the rat liver

Paraoxon infused into the rat liver during perfusion in situ with Waymouth's medium underwent chromatographic translobular migration with an apparent hepatic transit time of 3 min. Intralobular heterogeneity of paraoxon metabolism was examined by analyzing metabolites produced under conditions minimizing the chromatographic translobular migration of paraoxon. Periportal and centrilobular activities were estimated following forward and retrograde infusion of paraoxon, respectively. Centrilobular hepatocytes exhibited nearly twice the metabolic rate of the periportal cells. Pretreatment of the rat with DDE resulted in about a threefold increase in the ratio of oxidative deethylation to hydrolytic dearylation in the centrilobular region. The differentials observed by these analyses were less pronounced than expected from enzyme analyses in vitro, possibly reflecting secondary metabolism or intracellular heterogeneity of metabolic activities.     

Rat liver paraoxonase (paraoxon arylesterase)

Paraoxonase in the liver of male Sprague-Dawley rats was studied by using [phenyl-1-¹⁴C]paraoxon. Examination of the enzyme activity in subcellular fractions of liver homogenates indicated that hepatic paraoxonase is essentially a microsomal enzyme with a pH optimum of 7.5 to 7.8. Effects of calcium ions and EDTA on the enzyme suggested that active paraoxonase is a protein-calcium complex possibly with a range of affinity to calcium ion. Activity in homogenates declined with a half-life of 6 to 9 hr when stored at 0°C, apparently reflecting dissociation of calcium ions. Experiments with homogenates of perfused liver provided evidence that even without the contribution of calcium from blood, paraoxonase is almost fully active at the moment of homogenization. Possible reasons for the much reduced activity of paraoxonase in in vivo metabolism are discussed.     

Protective activity of the purified protein antigen of Erysipelothrix rhusiopathiae in pigs

We purified the protein antigen (P64), which contains 66 and 64 kDa proteins, from the alkaline extract (AE) of whole cells of Erysipelothrix rhusiopathiae strain Agata (serovar 5) to determine the protective activity of the antigen against E. rhusiopathiae infection in pigs. The serum titre of antibody against P64 rapidly increased in pigs immunized with 500 and 100 micrograms of P64 and reached maximum values at 3 weeks after the first immunization (1 week after the second immunization). However, the serum antibody titres were not increased in pigs immunized with 20 micrograms of P64 and in nonimmunized pigs. In the pigs immunized with live cell vaccine (acriflavin-fast attenuated strain Koganei 65-0.15), the serum titres of antibody against P64 also increased at 1-2 weeks after immunization. In a pig challenge test performed on immunized and nonimmunized pigs, all nonimmunized pigs showed typical clinical signs of swine erysipelas (fever, erysipeloid, arthritis), while all pigs immunized with 500 and 100 micrograms of P64 and live cell vaccine showed no clinical signs of this disease. In Western blot analysis, sera from pigs immunized with P64 and live cell vaccine strongly reacted with the 64 kDa protein. In contrast, the serum from nonimmunized pigs did not react with any proteins. From these results, it was suggested that a specific antibody against the 64 kDa protein could be increased in pigs immunized with P64 or live cell vaccine and that this anti-P64 antibody has a strong protective effect against E. rhusiopathiae infection in pigs.     

Preparation and partial characterization of monoclonal antibodies against the protective protein antigen of Erysipelothrix rhusiopathiae

Thirteen mouse monoclonal antibodies (Mabs) against the protective protein antigen (P64) of Erysipelothrix rhusiopathiae were prepared and partially characterized. The titres of the Mabs varied from 200 to 1,638,400 as determined by enzyme-linked immunosorbent assay (ELISA). Of the 13 Mabs 10, two and one belonged to the IgG2a, IgG1 and IgM subclasses, respectively. All Mabs reacted strongly with the 64 kDa protein and weakly with the 43 kDa protein upon Western blotting of the alkaline extract (AE) of E. rhusiopathiae. The protective activity (PD50/ml) of the 13 Mabs against E. rhusiopathiae infection in mice varied from < 50 to > 50,000. These Mabs were classified into three groups, highly protective Mabs, moderately protective Mabs and Mabs which did not possess protective activity, based on the protective index (ratio of the PD50/ml to the antibody titre). These results suggest that the 64 kDa protein is an effective protective antigen, which is easily cleaved into many small proteins, including the 43 kDa protein, and possesses at least two epitopes related to its protective activity and at least one epitope which is not related to protection of mice against E. rhusiopathiae infection.     

Effects of correcting missing daily feed intake values on the genetic parameters and estimated breeding values for feeding traits in pigs

Daily feed intake (DFI) is an important consideration for improving feed efficiency, but measurements using electronic feeder systems contain many missing and incorrect values. Therefore, we evaluated three methods for correcting missing DFI data (quadratic, orthogonal polynomial, and locally weighted (Loess) regression equations) and assessed the effects of these missing values on the genetic parameters and the estimated breeding values (EBV) for feeding traits. DFI records were obtained from 1622 Duroc pigs, comprising 902 individuals without missing DFI and 720 individuals with missing DFI. The Loess equation was the most suitable method for correcting the missing DFI values in 5–50% randomly deleted datasets among the three equations. Both variance components and heritability for the average DFI (ADFI) did not change because of the missing DFI proportion and Loess correction. In terms of rank correlation and information criteria, Loess correction improved the accuracy of EBV for ADFI compared to randomly deleted cases. These findings indicate that the Loess equation is useful for correcting missing DFI values for individual pigs and that the correction of missing DFI values could be effective for the estimation of breeding values and genetic improvement using EBV for feeding traits.