Effect of total mixed ration silage containing agricultural by-products with the fermented juice of epiphytic lactic acid bacteria on rumen fermentation and nitrogen balance in ewes

The goal of this study was to determine the effects of the fermented juice of epiphytic lactic acid bacteria (FJLB) on the quality of total mixed ration (TMR) silage containing agricultural by-products, its digestibility, rumen fermentation, and nitrogen balance in ewes. TMR was prepared from rice straw, corn stover silage, brewer grain, tofu waste, steam-flaked corn, and a mineral mixture. The treatments consisted of silage additives added to TMR: CON (no silage additive), FJLB, COM (commercial additive), and MIX (FJLB + COM). Four cannulated ewes were assigned to the 4 × 4 Latin square design. The MIX treatment produced a lower (P < 0.01) pH than did the CON and FJLB treatments and a higher (P < 0.01) lactic acid concentration than did the other treatments. The fiber content in the COM treatment was lower (P < 0.05) than that in the other treatments. The FJLB treatment had similar fermentation quality and chemical composition to those of the CON and COM treatments in all parameters observed. Although the silage quality index (Fleig point) was higher in the MIX and COM treatments than in the CON treatment, all silages had good quality. No silage additives affected intake, digestibility, rumen fermentation, or nitrogen balance. In conclusion, the TMR silage prepared from agricultural by-products mixed with wet-type food by-products with or without FJLB added resulted in well-preserved fermentation, and this product might be used as a ruminant feed.

     

Identification of a 4-Deoxy-l-erythro-5-hexoseulose Uronic Acid Reductase,FlRed, in an Alginolytic Bacterium Flavobacterium sp. Strain UMI-01

In alginate-assimilating bacteria, alginate is depolymerized to unsaturated monosaccharide by the actions of endolytic and exolytic alginate lyases (EC 4.2.2.3 and EC 4.2.2.11). The monosaccharide is non-enzymatically converted to 4-deoxy-l-erythro-5-hexoseulose uronic acid (DEH), then reduced to 2-keto-3-deoxy-d-gluconate (KDG) by a specific reductase, and metabolized through the Entner–Doudoroff pathway. Recently, the NADPH-dependent reductase A1-R that belongs to short-chain dehydrogenases/reductases (SDR) superfamily was identified as the DEH-reductase in Sphingomonas sp. A1. We have subsequently noticed that an SDR-like enzyme gene, flred, occurred in the genome of an alginolytic bacterium Flavobacterium sp. strain UMI-01. In the present study, we report on the deduced amino-acid sequence of flred and DEH-reducing activity of recombinant FlRed. The deduced amino-acid sequence of flred comprised 254 residues and showed 34% amino-acid identities to that of A1-R from Sphingomonas sp. A1 and 80%–88% to those of SDR-like enzymes from several alginolytic bacteria. Common sequence motifs of SDR-superfamily enzymes, e.g., the catalytic tetrad Asn-Lys-Tyr-Ser and the cofactor-binding sequence Thr-Gly-x-x-x-Gly-x-Gly in Rossmann fold, were completely conserved in FlRed. On the other hand, an Arg residue that determined the NADPH-specificity of Sphingomonas A1-R was replaced by Glu in FlRed. Thus, we investigated cofactor-preference of FlRed using a recombinant enzyme. As a result, the recombinant FlRed (recFlRed) was found to show high specificity to NADH. recFlRed exhibited practically no activity toward variety of aldehyde, ketone, keto ester, keto acid and aldose substrates except for DEH. On the basis of these results, we conclude that FlRed is the NADH-dependent DEH-specific SDR of Flavobacterium sp. strain UMI-01.     

Establishment of testicular and ovarian cell lines from Honmoroko (Gnathopogon caerulescens)

We succeeded to establish cell lines from endemic fish species Honmoroko Gnathopogon caerulescens, which inhabits Lake Biwa, the third oldest lake in the world. Two cell lines designated as RMT1 and RMO1 were established from testis and ovary of G. caerulescens, respectively. These cell lines were initially cultured in Leibovitz’s L-15 medium supplemented with fetal bovine serum (FBS), fish embryo extract, epidermal growth factor, and basic fibroblast growth factor. Further addition of forskolin and β-mercaptoethanol was required to establish and maintain these cell lines for more than 60 passages. RMT1 and RMO1 cells showed fibroblast- and epithelial-like morphology, respectively. From immunocytochemical staining and gene expression patterns, RMT1 cells showed a characteristic of testicular Sertoli cells and RMO1 cells did that of ovarian theca cells. Both RMT1 and RMO1 cells multiplied well in the medium supplemented with 10 % FBS at 28 °C and their minimum population doubling times were 24.4 and 28.8 h, respectively. At the 45th passage, most of the RMT1 and RMO1 cells had a hyperploid set of chromosomes (67.3 and 96.1 %, respectively). Cells with normal diploid chromosome set were not observed. RMT1 cells were transfected with an enhanced green fluorescent protein (EGFP) expression vector and human elongation factor 1 α promoter worked efficiently to express EGFP. In addition, EGFP-expressing cell lines were also established, suggesting that the cell lines could be utilized as an in vitro monitor system (biosensor) for the evaluation of endocrine disruptors which might affect gonadal function.     

Nonrandom distribution of chum salmon stocks in the Bering Sea and the North Pacific Ocean estimated using mitochondrial DNA microarray

More than 1,000 age-identified chum salmon Oncorhynchus keta collected at 23 stations in the Bering Sea and the North Pacific Ocean in June to July 2003 were used to estimate their origin of stocks using a DNA microarray developed for analyzing the mitochondrial (mt)DNA haplotypes. The observed haplotype distribution was nearly the same as that reported previously for fish collected in September 2002 and 2003 in the present surveyed areas. A conditional maximum-likelihood method for estimation of stock compositions indicated that the Japanese stocks mainly distributed in north central Bering Sea, whereas the Russian stocks were mainly in western Bering Sea. The North American stocks were abundant in eastern Bering Sea and around the Aleutian Islands. Such an area-specific stock composition was not significantly different between mature and immature fish. Thus, the combined results of 2 years suggest that the distribution of chum salmon is nonrandom in the surveyed areas in summer and autumn, and that fish of the same origin migrate together to the same area irrespective of age.     

Relationship between roughness parameters based on material ratio curve and tactile roughness for sanded surfaces of two hardwoods

The roughness parameters on the material ratio curves were related to tactile roughness for samples of buna and mizunara. The surfaces of the samples were sanded using various grades of coated abrasives and the roughness parameters, reduced peak height (Rpk), core roughness depth (Rk), and reduced valley depth (Rvk), were estimated on the material ratio curves, which were obtained from roughness profiles determined using robust Gaussian regression filter. The values of Rpk and Rk were almost the same for buna and mizunara under the same sanding conditions and increased exponentially with tactile roughness. The coefficients of determination of those parameters and tactile roughness were higher than 0.79 at all cutoff wavelengths. On the other hand, the value of Rvk for mizunara was significantly larger than that for buna because of the deep local valleys. There was no relationship between Rvk and tactile roughness for both species.     

Carrier rate of Factor XI deficiency in stunted Japanese black cattle

Blood examinations and genotyping of Factor XI (F11) were performed in growth retardation Japanese Black cattle and their dams. Genotyping of F11 revealed that the recessive homozygous and heterozygous genotype frequencies were 5.2% and 50.0% in the Claudin-16 (CL-16) deficiency group (n=58), 0% and 14.2% in the renal dysplasia group (n=7), 0% and 26.1% in the non-CL-16 deficiency nephritis group (n=23), 8.9% and 46.7% in the hypogenesis syndrome group (n=45), 6.2% and 25.0% in the neonatal weak calf syndrome group (n=32), 9.1% and 38.6% in the respective dams group (n=44), 0% and 23.1% in the normal cattle group (n=13), and 5.9% and 38.2% in total (n=222), respectively. These results showed that the carrier rate of F11 deficiency was high in Japanese Black cattle, and that the CL-16 deficiency, hypogenesis syndrome, neonatal weak calf syndrome, and dams groups had a large amount of recessive homozygous genotype than the other groups. No abnormal bleeding was observed clinically in the present study, and 4 of the recessive homozygous dams showed normal growth and parturition.     

Antimicrobial susceptibility of Escherichia coli isolated from feces of wild cranes migrating to Kagoshima, Japan

Susceptibility to 13 antimicrobial agents was examined for 138 Escherichia coli isolates obtained from 192 fecal samples of wild cranes that migrated for wintering to the Izumi plain, Kagoshima prefecture in Japan. The numbers of isolates that were resistant to the antimicrobials used in this study are as follows: oxytetracycline (OTC), 22 isolates; minocycline, 7 isolates; ampicillin (ABPC), 4 isolates; nalidixic acid, 4 isolates; enrofloxacin, 2 isolates; kanamycin, one isolate. Multidrug resistant isolates exhibiting 2-4 drug resistances were obtained. All of the OTC-resistant isolates carried either the tet (A) or tet(B) gene. The bla(TEM) gene was found in all of the ABPC-resistant isolates.     

Effect of SCD and SREBP genotypes on fatty acid composition in adipose tissue of Japanese Black cattle herds

Fatty acid composition of beef adipose tissue is one of important traits because high proportions of monounsaturated fatty acid are related to favorable beef flavor and tenderness. In this study, we investigated effects of genetic factors such as stearoyl-CoA desaturase (SCD) and sterol regulatory element binding protein (SREBP) on beef carcass traits including fatty acid composition using two cattle populations. Sire effect was significantly related to almost all traits except BMS, suggesting that the trait examined in this study is highly controlled by genetic factors. The effect of SCD genotype on fatty acid composition was detected remarkably in both cattle groups, especially on stearic and oleic acids. This result was consistent with our previous studies and suggests that SCD is associated with fatty acid composition. Unlike SCD genotyping, the effect of SREBP genotype was not identified in this study. Our results suggested that SCD genotype would contribute to improving beef quality in field populations. Further studies about the relationship among these factors will bring an insight into the molecular mechanism of fatty acid metabolism in cattle.     

An attempt at estrus detection in cattle by continuous measurements of ventral tail base surface temperature with supervised machine learning

We aimed to determine the effectiveness of estrus detection based on continuous measurements of the ventral tail base surface temperature (ST) with supervised machine learning in cattle. ST data were obtained through 51 estrus cycles on 11 female cattle (six Holsteins and five Japanese Blacks) using the tail-attached sensor. Three estrus detection models were constructed with the training data (n = 17) using machine learning techniques (random forest, artificial neural network, and support vector machine) based on 13 features extracted from sensing data (indicative of estrus-associated ST changes). Estrus detection abilities of the three models on test data (n = 34) were not statistically different among models in terms of sensitivity and precision (range 50.0% to 58.8% and 60.6% to 73.1%, respectively). The relatively poor performance of the models might indicate the difficulty of separating estrus-associated ST changes from estrus-independent fluctuations in ST.