Effects of Yuzu (Citrus junos) Peel from Waste as an Aquaculture Feed Supplement on Growth,Environmental Load,and Dark Muscle Discoloration in Yellowtail Seriola quinqueradiata

Beneficial utilization of yuzu (Citrus junos) peel was investigated to improve the commercial value of cultured yellowtail fish. The antioxidant effects of grated yuzu peel from pomace (yuzu paste) on prevention of dark muscle discoloration during storage in sliced yellowtail fish were tested. Four diets were prepared, containing 0, 10, 50, and 100 g yuzu paste in a 1,450-g diet (wet basis). Fish were fed one of the four diets for 10 weeks. Growth performance was not significantly different among the dietary groups, but fish fed a diet containing 100-g yuzu paste showed the least average body weight and a significant increase in fecal amount and fat levels in the feces, which suggest low digestibility and low absorption. Dark muscle discoloration was significantly reduced in all yuzu dietary groups compared to the control group during storage at 4°C. The optimal supplementation of yuzu paste in the fish diet is very effective at maintaining fish flesh freshness as a finishing diet without causing additional phosphorus or nitrogen pollution of the sea and inhibiting fish growth. This study provides a new way to utilize waste yuzu peel, which also helps reduce waste from yuzu processing.     

Immunogenicity and safety of an attenuated Bordetella bronchiseptica vaccine in pigs

The immunogenicity and safety of an attenuated Bordetella bronchiseptica vaccine for swine atrophic rhinitis (AR) was evaluated in 22 hysterectomy-produced, colostrum-deprived pigs and 18 conventional pigs. None of 8 pigs inoculated at 7 days of age intranasally with greater than or equal to 3 X 10(5) colony-forming units (CFU) of vaccinal strain/pig and 2 of 5 pigs inoculated at 7 days of age intranasally with 3 X 10(4) CFU of the vaccinal strain/pig developed AR after intranasal challenge exposure with a virulent strain at postinoculation week (PIW) 3. The remaining 3 vaccinated pigs and 4 nonvaccinated pigs developed AR. Thirteen pigs were inoculated intranasally with 3 X 10(6) to 3 X 10(9) CFU of the vaccinal strain at 7 days of age. At PIW 12, the pigs were killed and necropsied. None of the pigs had clinical signs of AR and/or pneumonia. Virulence was studied by transmission of vaccinal strain through 3 serial growing passages on the nasal mucosa of a litter of hysterectomy-produced colostrum-deprived pigs. Inoculum (nasal swab samples from 2 pigs 4 days after inoculation with 10(8) CFU of vaccinal strain at 5 days of age) was inoculated into the nasal cavity of 2 nonvaccinated pigs. This procedure was repeated 3 times. After the 1st passage, the vaccinal strain was recovered on postinoculation day 4, but after postinoculation day 4, the vaccinal strain was not recovered until the end of the 3rd passage. Turbinate atrophy or pneumonia was not recognized in these inoculated pigs. The vaccinal strain provided immunogenicity without ill effects.     

Allometric equations for accurate estimation of above-ground biomass in logged-over tropical rainforests in Sarawak, Malaysia

Although allometric equations can be used to accurately estimate biomass and/or carbon stock in forest ecosystems, few have been developed for logged-over tropical rainforests in Southeast Asia. We developed allometric relationships between tree size variables (stem diameter at breast height (dbh) and tree height) and leaf, branch, stem and total above-ground biomass in two logged-over tropical rainforests with different soil conditions in Sarawak, Malaysia. The study sites were originally classified as mainly lowland dipterocarp forest and have been selectively logged in the past 20 years. In total, 30 individuals from 27 species were harvested to measure above-ground parts. The correlation coefficients for the allometric relationships obtained for total above-ground biomass as a function of dbh had high values (0.99), although the relationships for leaf biomass had a relatively low coefficient (0.83). We also found relatively high coefficients for allometric relationships between tree height and plant-part biomass, ranging from 0.82 to 0.97. Moreover, there were no differences for allometric equations of total above-ground biomass between study sites. A comparison of equations of above-ground biomass in various previously reported tropical rainforests and pan-tropic general equations imply that our allometric equations differ largely from the equations for tropical primary forests, early successional secondary forest, and even for the general models. Therefore, choosing the biomass estimation models for above-ground biomass in the logged-over forests of Southeast Asia requires careful consideration of their suitability.     

Unique expression patterns of myosin heavy chain genes in the ductus arteriosus and uterus of rabbits.

In smooth muscle tissue, two smooth muscle myosin heavy chain (MHC) isoforms (SM1, SM2) and two non-muscle MHC isoforms (NMA, NMB) have been identified. The purpose of our study was to clarify whether smooth muscle MHC mRNA expression reflects the physiological and functional state of the muscle. We studied the expression pattern of MHC mRNAs, using the S1-nuclease mapping procedure, in functionally and morphologically changeable organs; the ductus arteriosus (DA) during development (25 and 29 days of gestation, and from 3-day-old neonates) and uteri from virgin, day-10 pregnant (P10) and day-29 pregnant (P29) rabbits. The results demonstrated that SM2 expression was greater in the fetal DA than in the fetal aortic and pulmonary arteries, but that it decreased significantly following closure of DA. In the gravid uterus, SM1 expression was significantly (P<0.05) strong compared to other MHC mRNAs from virgin to P10 rabbits. During pregnancy, NMB expression showed a tendency to increase until P10, and after P10, SM2 expression increased dramatically and NMB expression decreased to give almost a mirror image of the SM2 expression. Smooth muscle type (SM1, SM2) was significantly (P<0.05) strong compared to non-muscle type expression (NMA, NMB) at P29. These data suggest that smooth muscle MHC mRNA, especially SM2 expression reflects the physiological and functional state of the smooth muscle.     

Response of cholecystokinin and digestive enzyme mRNA levels to various feed ingredients in yellowtail Seriola quinqueradiata

Cholecystokinin (CCK) is the key regulator hormone that stimulates the secretion of digestive pancreatic enzymes in vertebrates. In fish, little is known about the mechanism of induction of CCK in the digestive tract by feed ingredients. To investigate the response of CCK and digestive enzymes to fish feed ingredients in yellowtail Seriola quinqueradiata, we performed a number of experiments in which we measured the mRNA levels of CCK, trypsin, and lipase after oral administration of a single bolus of the ingredients. We administered fish meal and fish oil in experiment 1; high and low concentrations of fish meal in experiment 2; and five different dietary protein sources (fish meal, soybean meal, soy protein concentrate, corn gluten meal, and glutamic acid fermentation by-products) in experiment 3. In experiments 1 and 3, only fish meal significantly increased the mRNA levels of CCK and digestive enzyme. In experiment 2, a high concentration of fish meal [20?% (w/v)] significantly increased CCK and trypsin mRNA levels, but a low concentration of fish meal [1?% (w/v)] did not. These results suggest that high concentrations of fish meal (the protein source in fish feed) has the most potent effect on stimulation of CCK synthesis and secretion of digestive enzymes in yellowtail.     

Purification and properties of a histidine decarboxylase from Staphylococcus epidermidis TYH1 isolated from Japanese fish-miso

Histidine decarboxylase (HDC) from Staphylococcus epidermidis TYH1, a halotolerant histamine-producing bacterium isolated from Japanese fermented fish-miso, was purified to homogeneity for the first time. The enzyme was purified 182-fold from cell-free extracts by ammonium sulfate precipitation, anion exchange chromatography and gel filtration chromatography. The N-terminal amino acid sequences of two polypeptide chains of 27–30 and 7–9 kDa were highly homologous with those of α- and β-chains of other staphylococcal HDCs. The optimum pH and temperature for the enzyme were 6.0 and 60 °C, respectively. This enzyme did not decarboxylate lysine, arginine, tyrosine, tryptophan or ornithine. The enzyme activity decreased with the addition of NaCl. At pH 4.8, the V _max and K _m values were 45.5 μmol histamine min^?1 mg^?1 and 1.10 mmol/L, respectively. Moreover, this enzyme was resistant to heat treatment (80 °C for 15 min) and was stable upon freezing at ?30 °C for 7 days. The very similar physiological properties of this enzyme and the almost identical N-terminal amino acid sequence to that of the HDC from S. capitis indicated that this enzyme may be evolutionally highly conserved in the genus Staphylococcus. The biophysical properties of staphylococcal HDC were elucidated using native purified enzyme.     

Regulatory network of hrp gene expression in Xanthomonas oryzae pv. oryzae

Like other plant-pathogenic bacteria, Xanthomonas oryzae pv. oryzae, the causal agent of bacterial leaf blight of rice, has hrp genes that are indispensable for its virulence. The hrp genes are involved in the construction of the type III secretion (T3S) apparatus, through which dozens of virulence-related proteins, called effectors, are directly secreted into plant cells to suppress and disturb plant immune systems and/or induce plant susceptibility genes. The expression of hrp genes is strictly regulated and induced only in plants and in certain nutrient-poor media. Two proteins, HrpG and HrpX, are known as key regulators for hrp gene expression. Great efforts by many researchers have revealed unexpectedly that, besides HrpG and HrpX, many regulators are involved in this regulation, some of which also regulate the expression of virulence-related genes other than hrp. Moreover, it has been found that HrpG and HrpX regulate not only hrp genes and effector genes but also genes unrelated to the T3S system. These findings suggest that the expression of the hrp gene is orchestrally regulated with other virulence-related genes by a complicated, sophisticated regulatory network in X. oryzae pv. oryzae.     

Effects of alcohol extract of defatted soybean meal on growth performance and digestive physiology of yellowtail <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis>

The effects of alcohol extract of soybean meal on growth performance, digestibility, and pancreatic enzyme and bile acid status of yellowtail Seriola quinqueradiata were studied. Fish were fed one of the following for 8 weeks: a fish meal (FM) control diet (FMD), a defatted soybean meal diet (SBMD), an alcohol-extracted SBM diet (ExSBMD), an ExSBMD with alcohol-extract diet (ExSBM + ExtD), and an FMD with alcohol-extract diet (FM + ExtD). Growth performance was significantly inferior in fish fed with SBM, ExSBM + ExtD, and FM + ExtD compared to FMD- or ExSBMD-fed fish. Total bile acid levels, and trypsin and lipase activities in the anterior intestine were significantly lower in fish fed with SBM, ExSBM + ExtD, and FM + ExtD than in fish fed with ExSBMD and FMD, despite similar values in gallbladder or pyloric caeca. Cholecystokinin mRNA levels of the former diet-fed fish were significantly lower than those of FMD- or ExSBMD-fed fish. Lipid and protein digestibility of fish fed with SBMD, ExSBM + ExtD, and FM + ExtD was significantly reduced in comparison with that of fish fed with FMD or ExSBMD. These findings indicate that the alcohol extract of soybean meal inhibited the secretion of bile acids and pancreatic enzymes by a decrease of cholecystokinin stimulation. This inhibition seemed to be responsible for a low growth performance through impairment of the assimilation of lipid and protein.     

Yellowtail insulin-like growth factor 1: molecular cloning and response to various nutritional conditions

Insulin-like growth factor 1 (IGF1) plays an important role in fish growth. This study investigated the IGF1 response to various nutritional conditions in yellowtail. First, we cloned 1,075 bp of yellowtail IGF1 cDNA, which codes for a protein of 185 amino acids (aa). This is composed of 44 aa for the signal peptide; 68 aa for the mature peptide comprising the B, C, A, and D domains; and 73 aa for the E domain. The mature yellowtail IGF1 showed high identity to IGF1 of other teleosts. Insulin-like growth factor 1 mRNA expression in the liver and white muscle was measured to observe the IGF1 response to various nutritional conditions, because the liver has the highest IGF1 expression and white muscle comprises the largest fraction of the fish body. Only white muscle IGF1 mRNA expression decreased significantly by 3 wk of fasting and recovered by refeeding. In subsequent feeding ratio (1%, 2%, and 3%/BW/d) experiments, significant correlations to growth were observed in white muscle IGF1 mRNA expression at 2- and 6-wk points and in hepatic IGF1 mRNA expression at 4 wk point. These data suggest that IGF1 expression both in hepatic and white muscle is important for somatic growth in yellowtail. Furthermore, white muscle IGF1 mRNA expression showed better responses to somatic growth and nutrition status in our two experiments than hepatic IGF1 mRNA expression.