Immunohistochemical study on the delayed progression of epithelial apoptosis in follicle-associated epithelium of rat Peyer's patch

It is well known that some caspases in apoptosis is involved in determinant of terminal differentiation and maturation of various cells. Our previous study ultrastructurally clarified the differentiation into M cells from immature microvillous epithelial cells and the redifferentiation from M cells to microvillous epithelial cells in the follicle-associated epithelium (FAE) of rat Peyer's patch. In this study, the difference of epithelial apoptosis between the FAE of Peyer's patch and intestinal villi was immunohistochemically investigated in rat jejunoileum. As a result, cleaved caspase-3 was limited to several epithelial cells at the tip of FAE, whereas almost all of the epithelial cells were cleaved caspase-3 positive in intestinal villi. Cleaved caspase-9 was detected only in a few exfoliating or exfoliated epithelial cells of both FAE and intestinal villi. Nuclear DNA-fragmentation was detected only in several epithelial cells of the tip of FAE, while it was expressed from the middle regions in the intestinal villi. The DNase I expression of the epithelial cytoplasm was much weaker in FAE than in intestinal villi. Bcl-x expression was restricted in the apical cytoplasms of epithelial cells in the FAE, whereas it was restricted in whole cytoplasms in villous epithelial cells. These findings suggest that the progression of the apoptotic process in the epithelial cells of FAE is later than in the intestinal villi, so that the possibility of epithelial differentiation might be remained in the FAE, unlike in the intestinal villi.     

Effects of Dietary Vitamin C on Blood Chemistry and Nonspecific Immune Response of Juvenile Red Sea Bream,Pagrus major

A trial was conducted to determine the effect of ascorbyl‐2‐monophosphate Na/Ca (AMP‐Na/Ca) on blood chemistry and nonspecific immune response of red sea bream juveniles. Test diets with three levels of AsA (free, 107, and 325 mg/kg diet) were fed to juvenile red sea bream (36.0 ± 1.3 g) two times a day for 3 wk. There were no significant differences in hematocrit, glucose, and blood urea nitrogen. Total cholesterol and triglyceride in plasma of fish fed AsA‐free diet was significantly (P < 0.05) higher than that of fish fed two other diets. There were no significant differences in serum albumin, total bilirubin, and total serum protein. Glutamyl oxaloacetic transaminase in serum of fish fed diets containing 107 and 325 mg of AsA were significantly (P < 0.05) lower than that of fish fed AsA‐free diet. Serum lysozyme activity (LA) of fish fed diets containing 107 and 325 mg of AsA were significantly (P < 0.05) higher than that of fish fed AsA‐free diet. There was no significant difference in mucus LA. The results mentioned above demonstrated that AMP‐Na/Ca is a bioavailable AsA source for red sea bream juveniles. Supplement of more than 107 mg AsA/kg in diets improved blood chemistry and nonspecific immune function of red sea bream juveniles.     

Can fermented soybean meal and squid by‐product blend be used as fishmeal replacements for Japanese flounder (Paralichthys olivaceus)?

An experiment was carried out to evaluate fermented soybean meal and squid by‐product blend (1:1) (FP) as replacement of fishmeal (FM) for Japanese flounder, Paralichthys olivaceus. Five isocaloric (19 kJ g^?1) diets were prepared by replacing 0 (FP0), 12 (FP12), 24 (FP24), 36 (FP36) and 48% (FP48) FM protein with FP. Triplicate groups of juveniles (mean weight of 3.9 g) were delivered the test diets for 8 weeks in a flow‐through sea water system. The results showed that there were no significant differences (P > 0.05) among the growth rates of fish fed FP0, FP12, FP24 and FP36 diets. Growth and nutrient utilization parameters were significantly reduced in fish fed FP48 diet. Although, whole body proximate composition of fish were not significantly affected by the dietary treatments compared to the control; methionine and phenylalanine contents were significantly decreased in FP48 group. Protein retention was also significantly decreased in the similar group of fish. Dietary treatments did not alter most of the plasma metabolites, while some of the health parameters were improved in the replacement groups. Results suggested that FP is a potential candidate for alternative protein ingredient in aquafeed and can replace 36% FM protein in the diet of Japanese flounder.     

Expression of HSP70 in response to heat-shock and its cDNA cloning from Mediterranean blue mussel

Expression of HSP70 in response to heat-shock was investigated at the protein and mRNA levels in Mediterranean blue mussel. Western and Northern blot analyses revealed that HSP70 was expressed following heat-shock in the mantle at both protein and mRNA levels, suggesting that gene expression of HSP70 is implicated in the cellular response to heat-shock stress in mussel. It was then attempted to clone HSP70 cDNA in order to determine the primary structure of mussel HSP70. As a result, two full-length cDNA encoding HSP70 were isolated from a cDNA library prepared from the heat-shocked mantle. The isolated cDNA consist of single open reading frames of 2067 bp and 1911 bp which encode proteins of 689 amino acids and 637 amino acids, respectively. Both HSP70 cDNA encode an ATPase do main, and a substrate-binding do main in addition to a Glu-Glu-Val-Asp (EEVD) peptide motif that is specific for cytosolic HSP70. These findings suggest that the cDNA clones obtained in the present study encode cytosolic HSP70.     

Phytotoxins from anaerobically decomposing wheat straw

Phytotoxicity was demonstrated in the aqueous extract of wheat (Triticum aestivum, L.) straw suspensions. When extracts were incubated under anaerobic conditions, the development of phytotoxicity was greater at 20°C than at 10°C. The toxic products formed during incubation depended upon the incubation medium. Acetic and butyric acids were the major toxins produced in liquid straw fermentations during the first 2 weeks. However, after that time the acids did not account for the total toxicity, suggesting formation of unidentified phytotoxins. Acetic, propionic and butyric acids were the toxins formed in sand culture. The development and accumulation of phytotoxins were favored where the sand-straw mixtures were water-saturated. The implications of these findings to wheat cultural practice is discussed.     

Description and phylogeny of Ceratomyxa anko sp. n. and Zschokkella lophii sp. n. from the Japanese anglerfish, Lophius litulon (Jordan)

Two new species of myxozoans from the Japanese anglerfish, Lophius litulon, are described using myxospore morphology and small subunit rDNA sequences. Ceratomyxa anko sp. n. is a parasite of the gall bladder and had a prevalence of 57%. Mature spores of C. anko sp. n. are arcuate to crescent shaped with valves tapering to rounded tips. A prominent sutural line runs centrally between the round adjacent polar capsules containing the polar filament coiled two to three times. Spore measurements: length 10.8 (9.7-11.9) microm, width 41.9 (36.9-47.2) microm, polar capsule diameter 4.6 (4.1-5.3) microm. Ceratomyxa anko sp. n. can be distinguished from other Ceratomyxa spp. due to its spore dimensions and shape. Zschokkella lophii sp. n. is a parasite of the urinary bladder and had a prevalence of 70%. Mature spores are ellipsoidal to semicircular with bluntly pointed ends. The sutural line is curved or sinuous and the valves have no discernable surface ornamentation. Two almost spherical polar capsules are located separately in the ends of the spore, opening in almost opposite directions and contain the polar filament with five coils. Spore measurements: length 20.1 (16.8-24.0) microm, width 14.9 (12.7-16.8) microm, polar capsule diameter 5.1 (3.6-5.8) microm. Zschokkella lophii sp. n. can be distinguished from other Zschokkella spp. due to the terminal opening of the polar capsules within the spores and the site of infection within the host fish. In the phylogenetic analyses, C. anko sp. n. grouped with other members of the same genus forming a monophyly. Zschokkella lophii sp. n. forms a discrete clade with another Zschokkella sp. that infects the urinary bladder of marine fish. This grouping forms a sister clade to one containing members of the genus Parvicapsula, all of which are parasites of the urinary system in marine fish.     

Synthesis and toxicological properties of some alkyl and aryl 3,4-methylenedioxyphenyl N,N′-thio-bis-N-methylcarbamates

A series of asymmetrical thio-bis-N-methylcarbamate derivatives were synthesised by combining 3,4-methylenedioxyphenyl N-methylcarbamate with different N-methylcarbamates. Some of these derivatives combined good insecticidal activity against the housefly with reduced acute oral toxicity to mice. The thio-bis-N-methylcarbamates obtained exhibited a lower anti-cholinesterase activity than the more toxic of the two carbamate moieties contained in each combination. The weak synergism of piperonyl butoxide on the insecticidal activity of the derivatives suggests the possibility of a self synergism phenomenon caused by the 3,4-methylenedioxyphenyl group.     

Effect of GnRH pretreatment on reproductive performance of postpartum suckled beef cows following synchronization of estrus using GnRH and PGF2alpha

The effect of GnRH pretreatment on estrus detection rate, precision of estrus, and reproductive performance of postpartum beef cows synchronized to estrus using GnRH and PGF2alpha was evaluated. In Exp. 1, Angus cows (n = 87) were randomly assigned by parity, postpartum interval, and body condition score (BCS) to receive either 1) GnRH on d -7 and PGF2alpha on d 0 (GP) or 2) the GP treatment and an additional injection of GnRH on d -16 (GGP). Estrus detection and AI were conducted twice daily from d -3 to d 3. At 72 h after PGF2alpha, all animals not previously detected in estrus were bred by AI and received a concurrent injection of GnRH (TAI). Synchronized pregnancy rates were numerically increased (P = 0.15) in cows treated with GGP (55%) compared with those on the GP treatment (44%). In Exp. 2, 1,276 spring-calving, suckled beef cows in nine herds were randomized to treatments as described for Exp. 1, except that the initial GnRH injection for the GGP treatment was administered on d -14. Herd affected all indicators of reproductive performance (P < 0.05). The percentage of animals detected in estrus prematurely (d -3 to d 0; 7%) was not affected by treatment. Estrus response rate was influenced by postpartum interval (< 60 vs > or = 60; 61 vs 73%; P < 0.01) and a three-way interaction of parity, BCS, and treatment (P < 0.01). Within animals with a BCS > or = 5.5, the GGP treatment tended to increase the detection of estrus in primiparous cows (GP vs GGP; 76 vs 91%; P = 0.11) and decrease detection in multiparous cows (GP vs GGP; 78 vs 72%; P < 0.10). However, because conception rate to TAI in animals with a BCS > or = 5.5 was greater (P < 0.05) in the GGP than in the GP group (28 vs 8%, respectively), this interaction was interpreted to represent a shift in interval to estrus induced by the GGP treatment, rather than a reduction in the synchronization of ovarian function. Conception rates of animals inseminated to an observed estrus did not differ among treatments (P = 0.15). Synchronized pregnancy rate tended (P = 0.06) to be greater in GGP- (53%) than in GP-treated animals (47%). In conclusion, pretreatment with GnRH tended to increase pregnancy rates during a 6-d synchronization period, primarily through enhanced conception rates of cows bred by TAI. In contrast to our hypothesis, GnRH pretreatment did not increase the percentage of animals detected in estrus or the precision of estrus expression.     

Influence of supplementary fibrolytic enzymes on the fermentation of corn and grass silages by mixed ruminal microorganisms in vitro

This study was done to determine the effectiveness of supplementary enzymes at increasing the fiber digestion by ruminal microorganisms and to assess whether enzyme activity limits the rate of fiber digestion in ruminal digesta. In vitro comparisons of enzyme activities in two feed enzyme preparations (A and B) with enzyme activities extracted from ruminal fluid indicated that the addition of fibrolytic enzymes at the application rates recommended by the manufacturers would not be expected to increase significantly glycanase and polysaccharidase activities in ruminal fluid. Preparations A and B both increased (P < 0.001) the rate of gas production from freeze-dried corn and grass silages in in vitro incubations with ruminal fluid, but only at concentrations much higher than recommended application rates. Autoclaved controls had little or no effect. Ultrafiltration of enzyme B indicated that most stimulation was due to components >100 kDa, which is consistent with the cause of the stimulation being enzyme activity. Fibrolytic enzymes from other sources were also able to stimulate gas production: increased rates of gas production were observed in seven out of eight combinations of "cellulase" and corn or grass silage (P < 0.05). The comparison of glycanase and polysaccharidase activities with gas-stimulatory activity in the different enzyme preparations indicated that the highest correlation was between increased gas production and enzyme activity against microgranular cellulose (P < 0.05). In a wider range of fibrolytic enzyme preparations, those with endo-(beta-1,4)- or exo-(beta-1,4)-xylanase activity equal to that of preparation A did not produce similar increased rates of fermentation of corn silage when glucanase activity was low (P > 0.05). In contrast, preparations with glucanase activity similar to enzyme A gave at least as great (P < 0.05) an improvement in gas production than enzyme A, irrespective of xylanase activity. It was concluded that enzyme activity, probably a type of endo-(beta-1,4)-glucanase activity, limits the rate of fermentation of corn and grass silage in the rumen. Enzyme supplements of the type used in these experiments are unlikely to possess sufficient activity to overcome this limitation by direct application to ruminal digesta, implying that treatment of the ration prefeeding will be key to harnessing the potential of exogenous fibrolytic enzymes in ruminant nutrition.