Expression and regulation of Foxa2 in the rat uterus during early pregnancy

The forkhead box a (Foxa) protein family has been found to play important roles in mammals. Recently, the expression of Foxa2 was reported in the mouse uterus, and it was reported to be involved in regulation of implantation. However, the regulation of Foxa2 expression in the uterus is still poorly understood. Therefore, the present study was conducted to investigate the expressional profiles of Foxa2 in the rat uterus during the estrus cycle and pregnancy. Furthermore, the effect of steroid hormones and Hedgehog protein on the expression of Foxa2 was analyzed in vivo and in vitro. In this study, the level of expression of Foxa2 was low in the rat uterus during the different stages of the estrus cycle. However, the expression increased transiently during early pregnancy at 3.5 days post coitus (dpc) and decreased at 5.5 dpc. In ovariectomized rats, P4 treatment had no effect on the expression of Foxa2 compared with the expression in control animals. Moreover, the expression of Foxa2 in cultured epithelial cells was not increased by P4 treatment in vitro. However, Foxa2 expression was significantly decreased in the rat uterus after 24 h of E2 treatment. Treatment of cells with a recombinant Hedgehog protein significantly increased the expression of Foxa2. These results suggest that the expression of Foxa2 may transiently increase just before the implantation and it may be regulated by E2 and Hedgehog protein.     

Three-dimensional reconstruction of intramuscular collagen networks of bovine muscle: A demonstration by an immunohistochemical/confocal laser-scanning microscopic method

We undertook a three‐dimensional reconstruction of intramuscular collagen networks of bovine muscle using an immunohistochemical/confocal laser‐scanning microscopic method. By immunohistochemical staining, type I and III collagens were observed mainly in the perimysium, while type IV collagen was observed in the endomysium. On the other hand, type V and VI collagens were observed in both the perimysium and endomysium. By confocal laser‐scanning microscopy, the collagen observed in the perimysium was three‐dimensionally reconstructed as plate‐shaped layers whereas the collagen observed in the endomysium surrounded myofibers. The three‐dimensionally reconstructed observations using immunohistochemical/confocal laser‐scanning microscopic method is useful for investigating collagen networks in muscle.     

An Endophytic Actinomycete, Streptomyces sp. AOK-30, Isolated from Mountain Laurel and Its Antifungal Activity

To survey endophytic actinomycetes as potential biocontrol agents against fungal diseases of mountain laurel, young plants of mountain laurel (Kalmia latifolia L.) were used as an isolation source. From a total of 73 actinomycetes isolates obtained from leaves, stems and roots of test plants, Streptomyces sp. AOK-30 was selected, because i) it had a broad and intense antimicrobial spectrum against various yeasts and fungal pathogens of Ericaceae, ii) it grew on the multiplication and rooting media for tissue culture for this plant, and iii) the tissue-cultured seedlings that had been treated with this isolate became resistant to Pestalotia disease without any adverse growth effects such as malformation, dwarfing, discolorization and defoliation. This isolate was identified as Streptomyces sp., based on cultural, physiological and morphological characteristics. The present results indicate that isolate AOK-30 is a potential biocontrol agent against diseases of mountain laurel. Received 10 September 2002/ Accepted in revised form 22 November 2002     

Effect of dietary Rhodobacter capsulatus on cholesterol, triglycerides concentration, and meat quality of finishing pigs

The study was designed to investigate the effect of Rhodobacter capsulatus on serum and meat cholesterol, fatty acid composition in meat, as well as meat quality of finishing pigs. A total of 16 120-day-old Landrace female pigs of about 60 kg initial body weight were randomly assigned into two groups. The pigs were fed a supplemented diet with 0.04% dietary R. capsulatus or a control diet for 60 days. Total cholesterol and low density lipoprotein-cholesterol concentration in serum was significantly lowered ( P  < 0.05) in the pigs fed the R. capsulatus supplemented diet compared to the control diet. Carcass weight, carcass length, shoulder fat, back fat and loin fat thickness, longissimus muscle (LM) area, and color score did not differ significantly between the pigs fed the R. capsulatus supplemented diet and control diet. Among the nutrient composition of LM meat, neutral fat and triglyceride concentration were significantly ( P  < 0.05) reduced in the finishing pigs by dietary supplementation of R. capsulatus . The proportions of n-6 PUFA were higher ( P  < 0.05) in the pigs fed the R. capsulatus supplemented diet than in the pigs fed the control diet. The supplementation of dietary R. capsulatus to finishing pig diet played important roles in reducing serum cholesterol and meat triglycerides, as well as in increasing polyunsaturated fatty acid content in LM meat.     

Comparative observation on the growth changes of the histochemical properties and collagen architecture of the Musculus puboischiofemoralis pars medialis from Silky, layer-type and meat-type cockerels

Growth‐related changes in the histochemical properties and collagen architecture of the Musculus puboischiofemoralis pars medialis were compared among Silky, layer‐type, and meat‐type cockerels. Histochemical and immunohistochemical methods were employed and collagen architecture was studied using scanning electron microscopy. Total intramuscular collagen was also determined. The myofibers were categorized as type I, type IIA and a transitional form, type I‐tr. The proportion of type I‐tr myofibers diminished and these myofibers were transformed entirely into type I myofibers in meat‐type but incompletely in the others. The largest diameter of type I myofiber was found in layer‐type at 30 weeks of age. At 30 weeks of age, layer‐type birds had attained well‐developed perimysial collagen bundles while meat‐type birds had less developed bundles. The endomysial collagen network had a large mesh size at 1 week and then accumulated many collagen fibrils to form a felt‐like fabric of fibrils by 30 weeks of age. Silky birds developed the thickest endomysial collagen plates of all the breeds. From these results it appears that growth‐related changes in the histological structure of M. puboischiofemoralis pars medialis are not necessarily causally affected by the different growth rates of chicken breeds.     

Bacteriocin production of probiotic Lactobacillus gasseri LA39 isolated from human feces in milk-based media

The use of bacteriocins from Lactobacillus gasseri , a probiotic lactic acid bacterium, as bio-preservatives in the food industry and animal formulations has been limited because few strains of Lb. gasseri are cultivated and produce a bacteriocin in natural media such as milk and milk-based media. By the determination of the growth-supplements to milk among the 47 nutrients, Lb. gasseri JCM1131^T, LA39 and LA158 isolated from human feces were successfully cultured in reconstituted skim milk and cheese whey using proteose peptone as a nutrient supplement, where Lb. gasseri LA39 produced a useful bacteriocin, gassericin A, with effective growth-inhibiting activity against Gram-positive food-borne pathogens. The data suggest these developed low-cost safe media supporting enough production of bacteriocins by the probiotic Lb. gasseri LA39 could be used to improve the safe bio-preservation of foods and therapy of bovine mastitis, and extra cheese whey produced by cheese making industry is reused in the cultivation for probiotics effectively.     

Distribution of taste buds in layer‐type chickens: Scanning electron microscopic observations

The distribution of taste buds of layer‐type chickens, from day 1 (H1) to day 140 (H140) post‐hatching, were investigated histologically and with a scanning electron microscope. Two types of openings were observed on the oral epithelium: large openings that were greater than 10 μm in width, and had an ellipsoid or spindle shape; and small openings that were less than 7 μm in width, and had a circular or polygon shape. It was defined that the former represent the openings of the salivary glands, and the latter are those of the taste buds. The number of taste buds remained constant from H1 to H140. Therefore, no significant change was observed between these two time points. To investigate the distribution of the taste buds in the whole oral cavity, a distribution map was created.     

Feed intake, lactation performance, blood metabolites and fertility in early lactation dairy cows grazing a timothy pasture

The present study was conducted to investigate feed intake, milk yield, milk composition, blood metabolites and fertility in early lactation dairy cows grazing a timothy pasture. Fourteen multiparous Holstein cows that calved between 20 May and 19 July were used over a 3‐year period. The stocking rate was 3.6–4.3 cow/ha. Concentrates were fed separately at 9.5–11.5 kg/day per cow (dry matter basis) from 1 to 13 weeks postparturition. Herbage intake was estimated using chromium oxide as an indigestible marker. The mean contents of crude protein, total digestible nutrients and neutral detergent fiber of pasture during the 3‐year study period were 22.3%, 71.8% and 51.7%, and those of total diet were 18.9%, 77.3% and 40.3%, respectively. The mean herbage dry matter intake was 13.0 kg/day from 2 to 13 weeks postparturition during the study, total dry matter intake was 23.7 kg/day, the total digestible nutrients sufficiency rate was 105%, milk yield was 39.7 kg/day, and milk fat percentage was 3.30%. The decrease in bodyweight postparturition was slight. Urea nitrogen concentrations in serum were below 18.3 mg/dL. The mean days to first estrus and days open were 36 and 104 days, respectively. These results indicate that energy deficiency, decrease in bodyweight and fertility in early lactation barely occur when high producing dairy cows are fed enough grazing grass and suitable concentrates.     

Detection of indoleamine 2,3-dioxygenase 1-expressing cells in canine normal and tumor tissues

Cancer immunotherapy is a novel cancer treatment for canine tumors. Indoleamine 2,3-dioxygenase 1 (IDO1) is overexpressed in some human tumors and inhibits antitumor immunity. In this study, we comprehensively evaluated expression pattern of IDO1 and the nature of IDO1-expressing cells in canine normal and tumor tissues. In normal tissue samples, IDO1 expression was detected only in the lymph nodes, spleen, tonsil tissues, and colon tissues. In contrast, IDO1-positive tumor cells were observed in several tumor tissue types. This is the first study to evaluate IDO1 expression in canine normal and tumor tissues, and the results suggest that IDO1 is a promising target for novel cancer immunotherapy in dogs with tumors.     

In vitro measurement of post‐natal changes in proliferating satellite cell frequency during rat muscle growth

Satellite cells, resident myogenic stem cells found in postnatal skeletal muscle, are most abundant during early postnatal development and sharply decline in frequency thereafter to adult levels in mice and rats. Therefore, postnatal changes in satellite cell mitotic activities are important aspects for further understanding a muscle growth strategy. In large meat‐production animals, however, the traditional in vivo proliferation assay may be less realistic because it requires intra‐peritoneal (ip) injection of huge dosage of mutagenic nucleosides, ³H‐labeled thymidine or bromodeoxyuridine (BrdU), at each age‐time of sacrifice. We report in the present pilot study using rats that in vivo proliferation activity of satellite cells can be evaluated by an in vitro BrdU‐incorporation assay in early cultures. Briefly, satellite cells were prepared from upper hind‐limb and back muscles and maintained for 24 h with imposing by BrdU addition for the last 2 h, followed by the regular immunocytochemistry for determining BrdU‐incorporated cell percentage. This in vitro assay demonstrated a rapid decrease in proliferating satellite cell frequency to the adult level during about 3‐month period after birth, and yielded a high correlation to the measurements by the in vivo BrdU ip‐injection method during the postnatal period examined from day‐2 to month‐11. The in vitro proliferation assay may be further adaptable for large domestic animals by the combination with a muscle biopsy technique that enables age‐interval sampling from the same growing animals.