Partial characterization and ontogenetic development of pancreatic digestive enzymes in Japanese eel Anguilla japonica larvae

The pancreatic digestive enzymes, trypsin, chymotrypsin, lipase and amylase were partially characterized, and changes in their activities were examined during the initial ontogeny of Japanese eel Anguilla japonica larvae from 5 to 34 days post-hatching (dph). The pH optima of the eel larval enzymes were narrower than those other fish species; trypsin activity was highest at pH 9, chymotrypsin and amylase activities were highest at pH 7 and 8, and lipase activity was highest at pH 8 and 9. In an analysis of thermal profiles, the larval pancreatic enzymes had a high optimal temperature and high thermal stability, which are typical of fish from the tropics. At 12 and 13 dph, lipase activity and gene expression levels of trypsin (-a and -b), lipase and amylase decreased markedly, suggesting a marked change in larval metabolism at that time. These data could be useful in the development of artificial larval diets in Japanese eel.     

Colonization of Pythium oligandrum in the tomato rhizosphere for biological control of bacterial wilt disease analyzed by real-time PCR and confocal laser-scanning microscopy

It recently has been reported that the non-plant-pathogenic oomycete Pythium oligandrum suppresses bacterial wilt caused by Ralstonia solanacearum in tomato. As one approach to determine disease-suppressive mechanisms of action, we analyzed the colonization of P. oligandrum in rhizospheres of tomato using real-time polymerase chain reaction (PCR) and confocal laser-scanning microscopy. The real-time PCR specifically quantified P. oligandrum in the tomato rhizosphere that is reliable over a range of 0.1 pg to 1 ng of P. oligandrum DNA from 25 mg dry weight of soil. Rhizosphere populations of P. oligandrum from tomato grown for 3 weeks in both unsterilized and sterilized field soils similarly increased with the initial application of at least 5 x 10(5) oospores per plant. Confocal microscopic observation also showed that hyphal development was frequent on the root surface and some hyphae penetrated into root epidermis. However, rhizosphere population dynamics after transplanting into sterilized soil showed that the P. oligandrum population decreased with time after transplanting, particularly at the root tips, indicating that this biocontrol fungus is rhizosphere competent but does not actively spread along roots. Protection over the long term from root-infecting pathogens does not seem to involve direct competition. However, sparse rhizosphere colonization of P. oligandrum reduced the bacterial wilt as well as more extensive colonization, which did not reduce the rhizosphere population of R. solanacearum. These results suggest that competition for infection sites and nutrients in rhizosphere is not the primary biocontrol mechanism of bacterial wilt by P. oligandrum.     

Growth performance and physiological condition of F1 amago salmon Oncorhynchus masou ishikawae juveniles obtained from broodstock with selective breeding for growth on a low fish-meal diet

A 10-week feeding trial was conducted to evaluate the growth and physiological condition of F1 amago salmon Oncorhynchus masou ishikawae juveniles obtained from broodstock that had been selected for growth on a low fish-meal (LFM) diet. F1 juveniles from broodstock showing superior or inferior growth on a LFM diet (LFM-S, LFM-I), and those from broodstock showing intermediate growth on a fish meal-based (FM) diet (FM-M), were fed a LFM diet and a FM diet. In fish fed the LFM diet, growth of LFM-S was significantly better than FM-M at 3 weeks; however, this superiority in growth disappeared at the end of the 10-week trial. Growth of LFM-I was significantly lower than LFM-S throughout the feeding trial. Growth of fish fed the FM diet was not significantly different among groups, and higher than fish fed the LFM diet. Improvements in the blood hemoglobin concentration and morphological conditions of the distal intestine and liver were observed in LFM-S fed the LFM diet. Although the effect of selective breeding was limited in F1 amago salmon, the present results suggest that continuous selection for growth with a LFM diet has potential as a method to adapt to the limited supply of fish meal.     

Studies on supplementary desalted mother liquor on digestibility of nutrients,ruminal fermentation,and energy and nitrogen balance in Thai native cattle

Four Thai native steers were used to determine the adequate levels of supplementary desalted mother liquor (DML) for energy and nitrogen balances and ruminal fermentation. The crude protein and sodium chloride contents of DML were 25.5% and 60.3% on a dry matter (DM) basis, respectively. A 4 × 4 Latin square design experiment was conducted by adding different amounts of DML to three experimental diets (T1: 1.1%, T2: 2.2%, T3: 3.4% sodium chloride concentration with supplementary DML on a DM basis) and comparing their effects with those of a control diet (C) containing 1.0% commercial salt on a DM basis. The animals were given the experimental diets and rice straw daily at 1.2% and 0.8% of body weight, respectively, on a DM basis. No significant differences in the apparent digestibility of nutrients were observed among treatments. T3 achieved the lowest nitrogen retention (P < 0.05), followed by C, T2 and T1. The ratios of energy retention to gross energy were higher in T1 and T3 than T2, and that in C was lowest (P < 0.05). Supplementary NaCl concentration at 1% and 2% can be replaced with DML without an adverse effect on the digestibility of nutrients or on the nitrogen and energy retention.     

Changes in the pH,EC, available P,SOC and TN stocks in a single rice paddy after long-term application of inorganic fertilizers and organic matters in a cold temperate region of Japan

Purpose

The objective of this study was to determine the changes in the main soil chemical properties including pH, electrical conductivity (EC), available phosphorus (P), soil organic carbon (SOC) and total nitrogen (TN) stocks after long-term (31 years) additions of two types of organic matters—rice straw and rice straw compost, combined with NPK fertilizers in single rice paddy in a cold temperate region of Japan.

Materials and methods

A long-term experiment on combined inorganic fertilizers and organic matters in paddy rice cultivation began in May 1982 in Yamagata, northeastern Japan. After the 31st harvest, soil samples were collected from five treatments [(1) PK, (2) NPK, (3) NPK + 6 Mg ha^?1 rice straw (RS), (4) NPK + 10 Mg ha^?1 rice straw compost (CM1), and (5) NPK + 30 Mg ha^?1 rice straw compost (CM3)] at five soil depths (0–5, 5–10, 10–15, 15–20, and 20–25 cm). Soil chemical properties of pH, EC, available P, SOC, and TN were analyzed.

Results and discussion

The pH decreased significantly only at the higher compost rate of 30 Mg ha^?1, while EC increased in all the organic matter treatments. Available P significantly increased in the CM1 and CM3 treatments by 55.1 and 86.4 %. The amounts of SOC stock increased by 67.2, 21.4, and 8.6 %, and soil TN stock by 64.1, 20.2, and 8.5 % in CM3, RS, and CM1, respectively, compared to NPK treatment.

Conclusions

Significant changes in soil properties were observed after 31 years of organic matter applications with reference to PK- and NPK-fertilized rice paddy soils. A significant decrease in pH was observed with the application of a high rate (30 Mg ha^?1) of rice straw compost but not with the conventional rate of 10 Mg ha^?1. However, EC increased significantly relative to that of the PK- and NPK-fertilized plots in all the organic matter treatments. Available P significantly increased in the CM1 and CM3 treatments by 55.1 and 86.4 %. The amounts of SOC stock expressed as a percentage of total C applied to the soil were higher from 10 Mg ha^?1 compost (28.7 %) than that from 6 Mg ha^?1 rice straw (17.4 %), indicating a more effective soil organic C accumulation from rice straw compost than that from original rice straw.

     

Survey and molecular characterization of Cryptosporidium and Giardia spp. in owned companion animal, dogs and cats, in Japan

Compared with other countries, surveys of these parasites have been rarely performed in companion animals of Japan in spite of their significance for public health. Here, we investigated pet dogs and cats in Japan for the first time, and genetically analyzed the isolates to evaluate the risk of zoonotic infections. Seventy-seven fecal samples were collected from privately owned dogs and 55 samples from owned cats in Osaka city, Japan. Cryptosporidium oocysts were identified in 3/77 dogs (3.9%) and 7/55 cats (12.7%), and Giardia infection in 2/77 dogs (2.6%) and 1/55 cats (1.8%). Amplification of the target regions for genotyping was successful, Cryptosporidium isolates in dogs and cats were identified as C. canis and C. felis, respectively, and those of Giardia in dogs and cats were G. intestinalis Assemblages D and F. The discharge period of the oocysts varied within 3-16 weeks and that of the cysts was 12 weeks. To date, zoonotic types of both parasites have been identified in other animals in Japan, and further large-scale studies are needed to determine the distribution of zoonotic genotypes in these animals, especially those closely associated with humans.     

Induction of apoptotic lesions in liver and lymphoid tissues and modulation of cytokine mRNA expression by acute exposure to deoxynivalenol in piglets

Six 1-month-old piglets were intravenously injected with deoxynivalenol (DON) at the concentration of 1 mg/kg body weight, with three pigs each necropsied at 6 and 24 h post-injection (PI) for investigation of hepatotoxicity and immunotoxicity with special attention to apoptotic changes and cytokine mRNA expression. Histopathological examination of the DON-injected pigs revealed systemic apoptosis of lymphocytes in lymphoid tissues and hepatocytes. Apoptosis of lymphocytes and hepatocytes was confirmed by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method and immunohistochemical staining against single-stranded DNA and cleaved caspase-3. The number of TUNEL-positive cells in the thymus and Peyer''s patches of the ileum was increased at 24 h PI compared to 6 h PI, but the peak was at 6 h PI in the liver. The mRNA expression of interleukin (IL)-1β, IL-6, IL-18, and tumor necrosis factor (TNF)-α in the spleen, thymus and mesenteric lymph nodes were determined by semi-quantitative RT-PCR, and elevated expression of IL-1β mRNA at 6 h PI and a decrease of IL-18 mRNA at 24 h PI were observed in the spleen. IL-1β and IL-6 mRNA expressions increased significantly at 6 h PI in the thymus, but TNF-α decreased at 6 h PI in the mesenteric lymph nodes. These results show the apoptosis of hepatocytes suggesting the hepatotoxic potential of DON, in addition to an immunotoxic effect on the modulation of proinflammatory cytokine genes in lymphoid organs with extensive apoptosis of lymphocytes induced by acute exposure to DON in pigs.