Role of taurine deficiency in inducing green liver symptom and effect of dietary taurine supplementation in improving growth in juvenile red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis> fed non-fishmeal diets based on soy protein concentrate

This study was conducted to investigate the mechanism of green liver symptom induction and the effect of dietary taurine supplementation on growth performance in juvenile red sea bream fed non-fishmeal diets based on soy protein concentrate (SPC). Juvenile fish (initial BW 72 g) were fed for 20 weeks on SPC diets supplemented with taurine at levels of 0, 1.0, and 2.0%. In the taurine-unsupplemented SPC diet group, specific growth rate (SGR) and feed conversion ratio (FCR) were significantly inferior (P < 0.001), and incidence of green liver was observed in 70% of fish. In this group, hepatopancreatic and plasma taurine concentrations were lowest (P < 0.05), hepatopancreatic content of bile pigments was highest (P < 0.05), and osmotic tolerance of erythrocytes was inferior (P < 0.05) among the dietary treatment groups. Serum osmolality of all treatment groups was at similar levels. These physiological abnormalities as well as SGR and FCR were improved by dietary taurine supplementation. These results indicate that the mechanism for induction of green liver symptom is bile pigment overproduction due to increased hemolysis because erythrocytes become osmotically fragile due to dietary taurine deficiency. Taurine supplementation of SPC diets is essential for maintaining normal physiological condition and growth performance in juvenile red sea bream.     

Seasonal dynamics and yearly change in the abundance of metacercariae of <Emphasis Type="Italic">Parabucephalopsis parasiluri</Emphasis> (Trematoda: Bucephalidae) in the second intermediate host in the Uji–Yodo River,central Japan

An introduced pathogenic fish trematode, Parabucephalopsis parasiluri Wang, 1985 (Digenea: Bucephalidae), was studied to determine the seasonal fluctuation in its metacercariae abundance. The timing of the recruitment period of the metacercariae and the relationship between yearly fluctuation in metacercarial abundance and some environmental factors were also examined to develop an effective plan for disease control. Monthly changes in metacercarial abundance in the caudal fin of pale chub (Zacco platypus) and of the prevalence of the first intermediate host (golden mussel Limnoperna fortunei) suggest that P. parasiluri generally has an annual life cycle. The metacercarial abundance in chub increased when the water temperature fell below 7°C in winter, but the rapidity of the increase varied between 2007 and 2008. The fluctuations in the abundance of metacercariae in winter from 2001 to 2008 showed that mean flow rate and mean water level in January were negatively related to the infection level each year, and positively related to the mean water temperature. Based on these results, we propose a control plan for parabucephalopsiosis involving discharge control; that is, more than 50 m³/s (corresponding to 150 m³/s in the Yodo River) of water should be flushed from upstream dams for a period of at least one week starting on the day when the water temperature falls below 7°C.     

Iron-Enriched Bread with Karkade (Hibiscus sabdariffa) and Wheat Flour

Karkade (Hibiscus sabdariffa) was blended with wheat flour to make bread. When 0.5% Karkade was blended with wheat flour, maximum bread height and specific volume (cm³/g) were obtained (pH 4.8–5.0); these properties gradually deteriorated with increased Karkade. The pH of the bread crumb decreased with increased Karkade, reaching pH 3.35 when blended with 10% Karkade. The pH of Karkade alone was 2.5, which was adjusted to ≈5.0 by the addition of alkali just before blending with wheat flour and making bread. Control of the Karkade pH resulted in bread height and specific volume recovering to the original optimal levels. In addition, the deep reddish purple color (513 nm) of the bread crumb changed to a brownish color crumb. The Fe content was 0.14 mg of Fe/gram of dry crumb, or 6.22 mg of Fe/60 g of fresh bread when 5% Karkade was blended with wheat flour.     

Springiness of Pancake and Its Relation to Binding of Prime Starch to Tailings in Stored Wheat Flour

Wheat flours were stored at room temperature (15–25°C), 40, 60, 80, and 100°C for various times. The baking performance of these flours was then evaluated in terms of the springiness of pancakes (recovery from crushing). Baking performance improved with increased storage time at each temperature. Brabender Amylograph tests of the flours indicated that the onset temperature in viscosity decreased with increased storage time at each temperature. When the flours were fractionated by acetic acid (pH 3.5) with mortar and pestle, recoveries of the water-solubles and gluten fractions were unchanged, but recoveries of prime starch and tailings fractions changed remarkably with increased storage time. On the other hand, those changes were not observed when flours were fractionated with a Waring blender. The binding of prime starch to tailings was correlated significantly with baking performance.     

Establishment and characterization of primary canine hepatocellular carcinoma cell lines producing alpha-fetoprotein

A primary cultured cell line named CHKS was established from a hepatocellular carcinoma (HCC) of a dog showing a high level of serum alpha-fetoprotein (AFP). CHKS secreted a 66 KDD AFP into the growth medium regardless of the presence or absence of fetal bovine serum (FBS). Cloning CHKS with limiting dilution produced 4 clones, CHKS-1, -2, -3, and -4, which secreted 826, 471, 70, and less than 10 ng/ml, respectively, of AFP into the culture medium. In culture, these cell lines were similar in morphology and proliferation pattern to epithelial cells and positive to periodic acid-Schiff (PAS) staining. The presence of mRNA for canine albumin was demonstrated by nested PCR. The doubling times of the clone cell lines were 21, 45, 36, and 35 h, saturation densities 34, 18, 22, and 24 x 10(4)/cm(2), and plating efficiencies 18, 45, 46, and 45%, respectively. Chromosome analysis of these cell lines showed near triploidy. These results show that CHKS and its clones have hepatic cell functions and are useful for carcinogenetic and clinical studies of canine HCC.     

Performance of particleboard manufactured using air-injection press I: effects of air-injection press on preventing blowout of board manufactured from low-moisture particles

An air-injection press (AIP) was developed to prevent accidental blowouts of boards during production. In this study, the effects of the AIP on preventing blowouts were investigated by artificially creating a blowout-prone condition, and the press was shown to be effective in preventing blowouts. The modulus of rupture of the boards was almost constant irrespective of pressing time. Longer pressing time resulted in higher internal bond strength when pressed at 170 °C. The thickness swelling of the boards pressed at 170 or 190 °C was almost uniform irrespective of pressing time, and the manufactured boards showed performance similar to those manufactured with an ordinary press. The AIP prevented blowouts sufficiently even when the pressure of the injected air was reduced, and this reduction did not adversely decrease the performance of the boards. Air injection reduced formaldehyde emissions from the board.     

Relationships among steroid hormone levels in newborn piglets,birth weight,placental weight,vitality of offspring and litter size

Blood estrone sulfate (E₁S), estrone (E₁), estradiol (E₂) and progesterone (P₄) in newborn piglets were measured to clarify the relationships among birth and placental weight, vitality of offspring and litter size. First, the association between vital status (normal, weak and stillborn) from 165 newborn piglets of 18 litters and steroid concentrations; second, steroid concentrations from 152 newborn normal piglets and litter size; and third, steroid content in fetal placenta from 50 newborn normal piglets of six litters and litter size, were investigated. In the normal group, the birth and placental weight were significantly higher than those in the other groups. Blood E₁S levels in the stillborn group were significantly lower, whereas E₁, E₂ and P₄ were significantly higher compared to the normal group. Blood and placental E₁S levels in the small litter group were significantly higher than those in the other groups. However, there was no significant difference among the three litter size groups in the levels of steroid hormones in maternal blood. These results indicate that vitality of newborn piglets is related to E₁S concentration of neonate, to birth weight and placental weight. However, steroid hormone concentrations of newborn piglets were greatly affected by the number of littermates.     

Ontogenic and morphological study of gonadal formation in genetically-modified sex reversal XYPOS mice

Mammalian sexual fate is determined by the presence or absence of sex determining region of the Y chromosome (Sry) in the “bipotential” gonads. Recent studies have demonstrated that both male and female sexual development are induced by distinct and active genetic pathways. Breeding the Y chromosome from Mus m. domesticus poschiavinus (POS) strains into C57BL/6J (B6J) mice (B6J-XY^POS) has been shown to induce sex reversal (75%: bilateral ovary, 25%: true hermaphrodites). However, our B6N-XY^POS mice, which were generated by backcrossing of B6J-XY^POS on an inbred B6N-XX, develop as males (36%: bilateral testis with fertility as well as bilateral ovary (34%), and the remainder develop as true hermaphrodites. Here, we investigated in detail the expressions of essential sex-related genes and histological features in B6N-XY^POS mice from the fetal period to adulthood. The onsets of both Sry and SRY-box 9 (Sox9) expressions as determined spatiotemporally by whole-mount immunohistochemistry in the B6N-XY^POS gonads occurred 2–3 tail somites later than those in B6N-XY^B6 gonads, but earlier than those in B6J-XY^POS, respectively. It is possible that such a small difference in timing of the Sry expression underlies testicular development in our B6N-XY^POS. Our study is the first to histologically show the expression and ectopic localization of a female-related gene in the XY^POS testes and a male-related gene in the XY^POS ovaries. The results from these and previous experiments indicate that the interplay between genome variants, epigenetics and developmental gene regulation is crucial for testis development.     

The combined effect of clothianidin and environmental stress on the behavioral and reproductive function in male mice

Neonicotinoids, some of the most widely used pesticides in the world, act as agonists to the nicotinic acetylcholine receptors (nAChRs) of insects, resulting in death from abnormal excitability. Neonicotinoids unexpectedly became a major topic as a compelling cause of honeybee colony collapse disorder, which is damaging crop production that requires pollination worldwide. Mammal nAChRs appear to have a certain affinity for neonicotinoids with lower levels than those of insects; there is thus rising concern about unpredictable adverse effects of neonicotinoids on vertebrates. We hypothesized that the effects of neonicotinoids would be enhanced under a chronic stressed condition, which is known to alter the expression of targets of neonicotinoids, i.e., neuronal nAChRs. We performed immunohistochemical and behavioral analyses in male mice actively administered a neonicotinoid, clothianidin (CTD; 0, 10, 50 and 250 mg/kg/day), for 4 weeks under an unpredictable chronic stress procedure. Vacuolated seminiferous epithelia and a decrease in the immunoreactivity of the antioxidant enzyme glutathione peroxidase 4 were observed in the testes of the CTD+stress mice. In an open field test, although the locomotor activities were not affected, the anxiety-like behaviors of the mice were elevated by both CTD and stress. The present study demonstrates that the behavioral and reproductive effects of CTD become more serious in combination with environmental stress, which may reflect our actual situation of multiple exposure.     

Improvement of dimensional stability and weatherability of composite board made from water-vapor-exploded wood elements by liquefied wood resin impregnation

High-density and high-resin-content boards were produced by phenolic resin impregnation into board materials prepared by the water-vapor-explosion process (WVE) to develop high-durability wood composite boards for exterior use. Wet-dry cyclic tests and accelerated weathering tests were conducted, and the fundamental properties were determined to examine the effect of resin impregnation on board qualities. Bending and internal bond strength of resin-impregnated boards (I-board) satisfied the criterion for 18-type particleboard described in JIS A 5908. Thickness swelling (TS) after 24-h water immersion was approximately 2%. Resin impregnation improved the dimensional stability of the boards. In wet—dry cyclic testing, TS of I-board was the same as that of plywood. The retention ratio of modulus of rupture of I-board was large; thus, I-board had high bond durability. Color change of I-board was less than that of ordinary particleboard after a 500-h accelerated weathering test. I-Board had lower surface roughness than boards produced by a spray application method (S-board) and higher water repellency, although the difference in resin contents of the face layer was small. Thus, it is suggested that the surface properties and weatherability of I-board were improved by impregnation of phenolic resin. High-density and resin-impregnated boards made from the WVE elements are expected to withstand actual exterior use. Part of this report was presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004