Functional role of type VI collagen during early feather development of the chicken embryo in vitro

The regulatory function of type VI collagen during early feather development in embryonic chickens was investigated at the cellular and organ levels. Immunohistochemical studies of embryonic chicken skin showed that type VI collagen was distributed in spatial‐specific and temporal‐specific manners related to early feather development. To clarify the role of type VI collagen, we studied the feather development in intact, reconstituted and reconstituted gel skin cultures. When ethyl‐3,4‐dihydroxybenzoate (EDHB) was added to the medium of intact skin as an inhibitor of type VI collagen synthesis, the feather buds did not elongate and the number of neural cell adhesion molecule (NCAM)‐positive cells was reduced. However, the magnitudes of both suppressive effects of EDHB were reduced by the addition of liquid type VI collagen. Similar improvement was also observed in the reconstituted skin with liquid type VI collagen and in the reconstituted gel skin with solid type VI collagen at a low concentration. Moreover, type VI collagen promoted feather bud development in the absence of EDHB. However, a high concentration of solid type VI collagen in the reconstituted gel skin arrested the feather bud elongation, and antitype VI collagen antibodies caused feather buds to become longer and smaller in the reconstituted skin. At the cellular level, type VI collagen affected the proliferation, migration and NCAM expression of mesenchymal cells. These results suggest that type VI collagen regulates early feather development by controlling mesenchymal cell behavior.     

Phytotoxic activity of pethoxamid in soil under different moisture conditions

The phytotoxic activity of soil-applied pethoxamid [2-chloro- N -(2-ethoxyethyl)- N -(2-methyl-1-phenyl-1-propanyl) acetamide], (TKC-94), on the plant growth of rice ( Oryza sativa cv. Kiyohatamochi ) seedlings as an assay plant in soil was investigated under different soil moisture conditions. The phytotoxic activity of pethoxamid mixed with soil on the shoot and root growth of rice seedlings was uppermost under the highest soil moisture condition and it decreased with declining soil moisture content, while the inhibition was greater on the root growth than the shoot growth. The amount of pethoxamid adsorbed on soil solid and the concentration of pethoxamid in soil water from soil applied with this herbicide were not influenced by the soil moisture content. In addition, the phytotoxic activity on the growth of rice seedlings in sea sand culture applied with the soil water from the herbicide-applied soil was not influenced by the soil moisture content. In the sea sand culture, the phytotoxic activity of pethoxamid was significantly reduced in negative water potential as the concentration of polyethylene glycol-6000 added to the water increased. It is suggested that the phytotoxic activity of pethoxamid in the soil primarily depends on the concentration in soil water, but the phytotoxic activity was affected by soil moisture through the effect on absorption of this herbicide by rice seedlings.     

Effects of lauric acid on physical, chemical and microbial characteristics in the rumen of steers on a high grain diet

The effects of being fed lauric acid on rumen characteristics were evaluated in a double 3 × 3 Latin square design using six Holstein steers with ruminal cannulas on a high grain diet. The steers were fed commercial concentrate (8.7 kg/day/steer) with one of three levels of lauric acid (0, 25 or 50 g/day/steer) and timothy hay (1.8 kg/day/steer). The feed intake and digestibility were determined. Ruminal fluid was collected at 3 h after feeding to determine chemical, physical and microbial parameters. An in vitro pure culture study was performed to determine the effects of lauric acid on Streptococcus bovis, a potent bloat‐ and acidosis‐promoting rumen bacterium. There were no differences in feed intake and digestibility among the treatments. The proportion of butyrate and the viscosity of the rumen fluid tended to be lowered (P < 0.08 and P < 0.09, respectively) and the stable ingesta volume increase was significantly decreased (P < 0.01) by the lauric acid feed. The abundance of protozoa and bacteria did not differ among the treatments. In the in vitro study, the growth of S. bovis was inhibited by the lauric acid (100 nmol/L) but it showed an adaptive growth to lauric acid in long‐term subculturing. The S. bovis that had adapted to lauric acid showed decreased viscosity and lactate production (P < 0.01) in culture with sucrose. These results indicate that supplemental lauric acid added to a high grain diet improves physical properties, possibly by altering the metabolic activity of S. bovis, and it may prevent the occurrence of feedlot bloat and acidosis in beef cattle.     

Synergistic fibrolysis in the rumen by cellulolytic Ruminococcus flavefaciens and non-cellulolytic Selenomonas ruminantium: Evidence in defined cultures

To investigate the rumen bacterial interaction between cellulolytic Ruminococcus flavefaciens and non‐cellulolytic Selenomonas ruminantium, fiber digestibility and fermentation products were determined in defined cultures consisting of these two species. Avicel, orchardgrass hay, rice straw and alfalfa hay were used as substrates for 72 h incubation to monitor digestibility, volatile fatty acids, succinate, lactate and bacterial number. In monoculture, R. flavefaciens digested the fiber sources at 21–32%, while S. ruminantium strains did not. When R. flavefaciens was cocultured with one of three different strains (GA192, S137 and S150) of S. ruminantium, fiber digestion exceeded the value recorded by R. flavefaciens alone. In particular, cocultures with S. ruminantium S137 showed significantly higher digestibility for all the fiber sources than R. flavefaciens alone (P < 0.05). Propionate production and growth of S. ruminantium was notable in all cocultures but not in monocultures. Succinate was accumulated in monoculture of R. flavefaciens, while the accumulation was not observed in cocultures. These results indicate that R. flavefaciens provides fiber hydrolysis products to S. ruminantium as growth substrates. In addition, S. ruminantium could activate R. flavefaciens by rapidly consuming the products. Such cross‐feeding between cellulolytic and non‐cellulolytic bacteria could enhance fiber digestion, although the extent of the enhancement may depend on strain combinations.     

Effects of supplemental lauric acid-rich oils in high-grain diet on in vitro rumen fermentation

A series of in vitro studies were performed to evaluate the effects of lauric acid (LA)‐rich oils on rumen fermentation with a high‐grain diet. Soy oil (SO) and palm oil (PO) as long‐chain fatty acid triglycerides, palm kernel oil (PKO), coconut oil (CO), powdered coconut oil (pCO) and coconut oil calcium salt (COCa) as medium‐chain LA‐rich oils were used as tested additives. Rumen fluid from steers fed high‐grain diet was incubated with ground corn with or without oil supplementation (2.0 g/L) for 6 h at 39°C to monitor rumen products. Methane production decreased, while hydrogen production increased on LA‐rich oils except COCa. All the LA‐rich oils increased total volatile fatty acids (VFA) production and molar proportion of propionate. Also, amylase activity in culture was higher when these oils were added. The most potent additives, pCO and free LA, were further tested to determine dose–response of rumen fermentation. Powdered coconut oil and LA altered rumen fermentation toward more propionate production by supplementation at 1.2 and 0.3 g/L, respectively. These results suggest that some LA‐rich oils and free LA could be used for improving rumen fermentation under high‐grain diet feeding conditions.     

Frequencies of bovine PrP gene polymorphisms in Holstein and Japanese Black bulls in Japan

We screened for Japanese Black and Holstein bull sire samples to detect single nucleotide polymorphisms (SNPs) involving animo‐acid substitutions in the bovine prion gene in the entire coding region of the PRNP gene. Although three silent SNPs were found, we could not detect any SNP with animo‐acid substitution. We also examined the polymorphism of the octapeptide repeat number in these samples. There was no homozygous bull with repeat number 5. The frequency of heterozygous (6/5) bulls was 8% in the Japanese Black bull and 4% in the Holstein bull, respectively. The bull samples used in this study contain popular elite sires, so it appears that the polymorphisms of prion protein (PrP) are rather difficult to find in these two breeds in Japan, except for polymorphism of the octapeptide repeat number.     

cDNA cloning and characterization of two gelatinases from Japanese flounder

SUMMARY: Toughness is one of the most important elements that define the commercial value of the raw meat of fish. Degradation of the extracellular matrix is thought to be a cause of postmortem tenderization of fish meat. A previous study has suggested that this tenderization is caused mainly by metalloproteinases. The present study seeks to identify the proteinase(s) involved in tenderization; hence, cloned cDNA of two gelatinases from Japanese flounder, which showed high homology with mammalian matrix metalloproteinase (MMP)-2 and MMP-9, were designated as jfMMP-2 and jfMMP-9 , respectively. Northern blot analysis revealed that jfMMP-2 mRNA was expressed almost ubiquitously in adult tissues including the brain, muscle, gill, heart, gut, kidney, spleen, testis, and ovary. In contrast, the expression of jfMMP-9 mRNA was observed in those tissues which were abundant in blood cells, such as kidney, spleen, heart, and gill. Both recombinant proteins (jfMMP-2 and jfMMP-9) produced with the COS-7 cell system exhibited gelatin-degrading activity that was sensitive to 1,10-phenanthroline, a typical metalloproteinase inhibitor.