Use of Mg-L-Ascorbyl-2-Phosphate as a Vitamin C Source in Shrimp Diets

To overcome the lability of vitamin C (L-ascorbic acid or L-AsA), an essential nutrient of shrimp diets, the efficacy of Mg-L-ascorbyl-2-phosphate (MAP), a relatively stable derivative of L-AsA, was examined.
Groups of juvenile Penaeus japonicus were fed five different test diets containing different amounts of MAP for 100 days, to determine if MAP could be utilized by shrimp. Normal growth was observed on test groups fed diets containing 1,078, 430 and 215 mg MAP per kg, whereas mass mortality occurred in populations fed diets containing 43 and 0 mg MAP per kg. The dead shrimp found in the latter two treatments exhibited blackened lesions underneath the exoskeleton. The symptom looked similar to what was reported in previous papers as L-AsA deficiency syndrome. The feeding experiment suggests that MAP can be utilized by P.japonicus as a suitable replacement for L-AsA, and a supplement of 215 to 430 mg in 1 kg is sufficient to prevent clinical signs of vitamin C deficiency.
Repeated measurements of remaining L-AsA and MAP, made after diet processing and storage, demonstrated that MAP was significantly more stable than L-AsA and was less prone to leaching in sea water.     

Development of myofibrillar ATPase assay system on pH stat

The ATPase assay system on pH stat apparatus was developed. For the ATPase activity measurement, hydrogen ion (H⁺) concentration delivered from inorganic phosphate (Pi) as a hydrolysis production of ATP was estimated by titrating with 20 mM NaOH solution instead of colorimetric measurement of Pi. Inclusion of 0.5 M KCl in the ATP stock solution and 2 mM Tris-maleate (pH 7.0) buffer in the reaction medium reduced the extent of pH change upon addition of ATP to initiate the ATPase reaction. The amount of H⁺ liberated from Pi was strongly affected by the set pH for the ATPase assay with a promotion at alkaline pH. Thus, it was required to estimate the coefficient to convert H⁺ to a Pi concentration at a specific pH. The specified coefficient at pH 7.0 was 1.248. ATPase assay on pH stat allowed us to follow the ATP hydrolysis by myofibrils continuously showing a curvature profile at low salt medium (≤0.2 M KCl) or a linear profile at high salt (≥0.3 M KCl).     

Estimation of beef marbling in the Longissimus muscle with computer image analysis of ultrasonic pictures of the Iliocostalis muscle area

This study investigated an objective method for estimating beef marbling using ultrasonic images of the Iliocostalis muscle and the Lomgissimus muscle area sections. Thirty‐one Japanese Black cattle steers were used in this study. The end of the left side shoulder blade bone was scanned using an ultrasonic device. Ultrasonic images were captured of the Longissimus muscle area and that around the Iliocostalis muscle area. Twenty items were measured in the two images using computer image analysis software. The level of beef marbling was measured according to the Beef Marbling Standard (BMS) for carcass grading, and the percentage of ether‐extractable fat content in the Longissimus muscle (EE). The difference in the gray level between the Iliocostalis muscle and intermuscular fat (X10) was used to estimate the BMS and the EE, which were highly correlated (r² = 67.72% and 61.30%). An equation was developed using four parameters from the two ultrasonic images, which could estimate the BMS (r² = 85.88%). This equation could also estimate the EE (r² = 68.98%). The equations used to estimate beef marbling were based on one to four parameters that included X10. Thus, ultrasonic images of the Iliocostalis muscle area section are important for estimating beef marbling.     

The Effects of Adjuvants on Autoimmune Responses Against Testicular Antigens in Mice

Experimental autoimmune orchitis (EAO) is a model of immunologic male infertility and pathologically characterized by lymphocytic inflammation, which causes breakdown of the testicular immune privilege with spermatogenic disturbance. Generally, murine EAO is induced by immunization with testicular homogenate (TH) from the testes of donor mice + complete Freund''s adjuvant (CFA) + Bordetella pertussigens (BP), and it has been considered that treatment with these two adjuvants is required to enhance the immune response against testicular antigens. However, there remains a possibility that CFA and BP may affect autoimmune responses against the testicular antigens without TH. In the present study, we examined this possibility using real-time RT-PCR, Western blotting and immunohistochemical staining. The results demonstrated that immunization with TH in combination with CFA and BP evoked more severe EAO than that with only TH. Real-time RT-PCR analyses revealed that Fas mRNA expression in TH+CFA+BP-induced EAO was significantly higher than that in TH-induced EAO. Interestingly, IL-6 mRNA expression dramatically increased in TH+CFA+BP-induced EAO; however, no apparent change in IL-6 mRNA expression occurred in TH-induced EAO. It was also noted that treatment with CFA and BP alone augmented autoimmune reactions against some testicular autoantigens. These results indicates that these adjuvants are helpful in evoking severe EAO, and treatment with the adjuvants alone can evoke autoimmune reactions against some testicular autoantigens despite the use of no TH.     

Species-specific thermal denaturation pattern of fish myosin when heated as myofibrils as studied by myosin subfragment-1 and rod denaturation rates

Thermal denaturation of myofibrils from various species of fish was investigated by measuring ATPase inactivation, myosin aggregation, myosin subfragment-1 (S-1) and rod denaturation rates as studied by chymotryptic digestion. Decrease in monomeric myosin (myosin aggregation) was always faster than the ATPase inactivation for all myofibrils tested. The relative denaturation rate of rod to that of S-1 differed from species to species. Preceded denaturation of rod was observed with some species, and the opposite was true with other species. The denaturation pattern was explained by the different magnitude of S-1 stabilization by F-actin in myofibrils at low salt medium. Myofibrils which receive a great stabilization by F-actin as studied by ATPase inactivation showed the preceded rod denaturation pattern, and vice versa. S-1 portion, not F-actin, determined the different stabilization of S-1 by F-actin in myofibrils.     

Disease resistance and hypocholesterolemia in yellowtail Seriola quinqueradiata fed a non-fishmeal diet

ABSTRACT:   The physiology of yellowtail fed a non-fishmeal diet was examined, with a specific interest in the role of taurine in disease resistance and cholesterol metabolism. Decrease of disease resistance in fish fed a non-fishmeal diet was confirmed by mortality due to natural infection with pseudotuberculosis and artificial infection with Lactococcus garvieae . It is suggested that the most important symptoms related to decrease of disease resistance in fish fed a non-fishmeal diet is anemia. Anemia was improved by supplementation with taurine. Significant elevation of relative expression of HMG-CoA reductase mRNA in fish fed a non-fishmeal diet suggests that cholesterol synthesis would be activated and not dysfunctional. Plasma cholesterol of these fish was elevated to the levels of control fish by supplementation of both cholesterol and taurine. These results suggest that hypocholesterolemia observed in fish fed a non-fishmeal diet compared with a fishmeal diet would be caused by insufficient dietary cholesterol and decrease of endogenous cholesterol due to the lack of dietary taurine.