Japanese Cattle Research Articles

Isolation of Epizootic Hemorrhagic Disease Virus Serotype 10 from Culicoides tainanus and Associated Infections in Livestock in Yunnan, China.

Two strains of viruses, JC13C644 and JC13C673, were isolated from Culicoides tainanus collected in Jiangcheng County, Yunnan Province, situated along the border area shared by China, Laos, and Vietnam. JC13C644 and JC13C673 viruses can cause cytopathic effect (CPE) in mammalian cells BHK21 and Vero cells, and cause morbidity and mortality in suckling mice 48 h after intracerebral inoculation. Whole-genome sequencing was performed, yielding complete sequences for all 10 segments from Seg-1 (3942nt) to Seg-10 (810nt). Phylogenetic analysis of the sub-core-shell (T2) showed that the JC13C644 and JC13C673 viruses clustered with the Epizootic Hemorrhagic Disease Virus (EHDV) isolated from Japan and Australia, with nucleotide and amino acid homology of 93.1% to 98.3% and 99.2% to 99.6%, respectively, suggesting that they were Eastern group EHDV. The phylogenetic analysis of outer capsid protein (OC1) and outer capsid protein (OC2) showed that the JC13C644 and JC13C673 viruses were clustered with the EHDV-10 isolated from Japan in 1998, with the nucleotide homology of 98.3% and 98.5%, and the amino acid homology of 99.6% and 99.6-99.8%, respectively, indicating that they belong to the EHDV-10. Seroepidemiological survey results demonstrated that JC13C644 virus-neutralizing antibodies were present in 29.02% (177/610) of locally collected cattle serum and 11.32% (89/786) of goat serum, implying the virus's presence in Jiangcheng, Yunnan Province. This finding suggests that EHDV-10 circulates not only among blood-sucking insects in nature but also infects local domestic animals in China. Notably, this marks the first-ever isolation of the virus in China and its discovery outside of Japan since its initial isolation from Japanese cattle. In light of these results, it is evident that EHDV Serotype 10 exists beyond Japan, notably in the natural vectors of southern Eurasia, with the capacity to infect local cattle and goats. Therefore, it is imperative to intensify the surveillance of EHDV infection in domestic animals, particularly focusing on the detection and monitoring of new virus serotypes that may emerge in the region and pose risks to animal health.

A rumen virosphere with implications of contribution to fermentation and methane production, and endemism in cattle breeds and individuals.

Viruses have a potential to modify the ruminal digestion via infection and cell lysis of prokaryotes, suggesting that viruses are related to animal performance and methane production. This study aimed to elucidate the genome-based diversity of rumen viral communities and the differences in virus structure between individuals and cattle breeds and to understand how viruses influence on the rumen. To these ends, a metagenomic sequencing of virus-like particles in the rumen of 22 Japanese cattle, including Japanese Black (JB, n = 8), Japanese Shorthorn (n = 2), and Japanese Black sires × Holstein dams crossbred steers (F1, n = 12) was conducted. Additionally, the rumen viromes of six JB and six F1 that were fed identical diets and kept in a single barn were compared. A total of 8,232 non-redundant viral genomes (≥5-kb length and ≥50% completeness), including 982 complete genomes, were constructed, and rumen virome exhibited lysogenic signatures. Furthermore, putative hosts of 1,223 viral genomes were predicted using tRNA and clustered regularly interspaced short palindromic repeat (CRISPR)-spacer matching. The genomes included 1 and 10 putative novel complete genomes associated with Fibrobacter and Ruminococcus, respectively, which are the main rumen cellulose-degrading bacteria. Additionally, the hosts of 22 viral genomes, including 2 complete genomes, were predicted as methanogens, such as Methanobrevibacter and Methanomethylophilus. Most rumen viruses were highly rumen and individual specific and related to rumen-specific prokaryotes. Furthermore, the rumen viral community structure was significantly different between JB and F1 steers, indicating that cattle breed is one of the factors influencing the rumen virome composition.IMPORTANCEHere, we investigated the individual and breed differences of the rumen viral community in Japanese cattle. In the process, we reconstructed putative novel complete viral genomes related to rumen fiber-degrading bacteria and methanogen. The finding strongly suggests that rumen viruses contribute to cellulose and hemicellulose digestion and methanogenesis. Notably, this study also found that rumen viruses are highly rumen and individual specific, suggesting that rumen viruses may not be transmitted through environmental exposure. More importantly, we revealed differences of viral communities between JB and F1 cattle, indicating that cattle breed is a factor that influences the establishment of rumen virome. These results suggest the possibility of rumen virus transmission from mother to offspring and its potential to influence beef production traits. These rumen viral genomes and findings provide new insights into the characterizations of the rumen viruses.

Prevalence and genetic diversity in bovine parechovirus infecting Japanese cattle

The first bovine parechovirus (Bo_ParV) was reported in 2021, and currently, only two nearly complete genome sequences of Bo_ParV are available. In this study, we detected Bo_ParVs in 10 out of 158 bovine fecal samples tested using real-time RT-PCR, and Bo_ParVs were isolated from three of these samples using MA104 cells. Analysis of the P1 region revealed that Bo_ParVs shared high pairwise amino acid sequence similarity (≥ 95.7% identity), suggesting antigenic similarity among Bo_ParVs, whereas nucleotide sequence identity values (≥ 84.8%) indicated more variability. A recombination breakpoint was identified in the 2B region, which may influence the evolution of this virus.

Complete genome constellations of two bovine rotavirus A strains isolated in Japan reveal a unique T9 NSP3 genotype.

Full genome sequencing of two bovine rotavirus A (RVA) strains isolated in Japan in 2019 revealed two genotype constellations; one had a constellation of G8-P[1]-I2-R2-C2-M2-A3-N2-T9-E2-H3. Thereupon, genotype T9 carried by RVA/Bovine-tc/JPN/AH1041/2022/G8P[1], constitutes a rare NSP3 genotype, and only two unusual Japanese bovine RVA strains have thus far been reported to carry this genotype. The other RVA/Bovine-tc/JPN/AH1207/2022/G6P[5] strain possessed a constellation of G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3. Phylogenetic analyses indicate that the majority of gene segments were most closely related to Japanese bovine RVAs, suggesting that both strains might have derived through multiple reassortment events from RVA strains circulating within Japanese cattle. The emergence of RVA strains in Japan and their reassortment with locally circulating atypical RVAs could have implications for current vaccination strategies.

Antimicrobial resistance and associated genetic background of Histophilus somni isolated from clinically affected and healthy cattle.

Histophilus somni, a member of the Pasteurellaceae family, causes various diseases, including thrombotic meningoencephalitis and respiratory diseases. Here, 166 isolates recovered from Japanese cattle with various diseases between the late 1970s and the 2010s were subjected to susceptibility testing against 14 antimicrobials (ampicillin, amoxicillin, cefazolin, ceftiofur, kanamycin, streptomycin, nalidixic acid, enrofloxacin, danofloxacin, florfenicol, erythromycin, tylosin, oxytetracycline, and fosfomycin). The proportions of antimicrobial-resistant/intermediate isolates were low in the total isolates, with resistance rates ranging from 0% for ceftiofur and florfenicol to 13.2% for ampicillin. However, relatively high minimum inhibitory concentrations (MICs) and resistance/intermediate rates were observed in the isolates from cattle with respiratory diseases; i.e., 21/53 isolates (39.6%) showed resistance or intermediate to one or more antimicrobials for treatment of respiratory diseases, and the resistance/intermediate rates to oxytetracycline, kanamycin, ampicillin, amoxicillin, nalidixic acid, and danofloxacin were 28.3, 24.5, 24.5, 13.2, 1.9, and 1.9%, respectively. Isolates with high MICs tended to possess antimicrobial resistance genes, which may confer antimicrobial resistance phenotypes. In particular, all isolates with MICs of ampicillin/amoxicillin, kanamycin, and oxytetracycline ≥2 μg/mL, ≥512 μg/mL, and ≥4 μg/mL possessed bla ROB - 1, aphA-1, and tetH/tetR, respectively, whereas isolates whose MICs were lower than the above-mentioned values did not possess these resistance genes. These results suggest that the resistance genes detected in this study are primarily responsible for the reduced susceptibility of H. somni strains to these antimicrobials. As integrative and conjugative element (ICEs)-associated genes were detected only in genetically related isolates possessing antimicrobial resistance genes, ICEs may play an important role in the spread of resistance genes in some genetic groups of H. somni strains.

Indigenous and Introduced: How Beef-eating Acceptance Affected the Cattle Industry in Modern Japan

The intention of this paper is to describe how the cattle industry in Japan had changed with the introduction of beef eating habits. Before the Japanese Meiji Restoration, meat-eating had been formally prohibited. In practice, some people had opportunities to eat game, although livestock had been severely tabooed. In the Civilization and Enlightenment period of Japan, beef eating spread rapidly. It was mainly eaten in an indigenous cooking method called a beef “hot pot.” This cooking method influenced the development of beef fattening. In the pre-war period, farmers kept only a single cow for plowing, transporting, and collecting fertilizer, but the process of fattening was added later introduced to satisfy people''s appetites for better-tasting beef. The goal of fattening had consistently been fatty meat. With the development of fattening technology, the ideal meat in the 1920s was fatty meat with intramuscular marbling. This beef-eating culture is inextricably linked with the fact that beef was first accepted as a hot pot dish in the modern era. Cattle fattening in developed areas took place within the conventional system of cattle rearing, which involved single cattle rearing system combined with arable agriculture. The fact that multiple cattle rearing based on the Western model did not take root is not seen as a delay in the modernization of Japanese livestock breeding, but rather as a technical and managerial rationale for combining single-head rearing practices with the utilization of that cattle for arable agriculture. Even though the indigenous cattle rearing system remained, the introduction of live cattle from colonial Korea brought complexities to the Japanese cattle industry. The importation of Korean cattle brought about a qualitative diversity as well as a quantitative expansion in terms of the supply of cheaper and leaner beef, and contributed to the change in consumption that has been termed the “popularization of meat eating.”

Comprehensive assessment of genetic diversity, structure, and relationship in four Japanese cattle breeds by Illumina 50 K SNP array analysis.

There are four unique cattle breeds in Japan: Japanese Black, Japanese Brown, Japanese Polled, and Japanese Shorthorn. The objective of this study was to comprehensively assess the genetic diversity, structure, relationship, and the degree of influence from foreign breeds (Angus, Simmental, Hanwoo, Shorthorn, Ayrshire, Brown Swiss, and Devon) in the Japanese cattle breeds using Illumina 50 K SNP array. In principal component analysis, each Japanese breed was separately clustered except for Japanese Shorthorn and Shorthorn. Japanese cattle breeds also showed different genetic components from each other at K ≥ 5 in population structure analysis. Japanese Shorthorn, on the other hand, had a very similar structure to Shorthorn at K ≤ 9, and Japanese Polled had a partially similar component with Angus at K = 3-7. Such close relationships were also observed in the phylogenetic tree. These findings imply that Japanese cattle breeds share genetic components with European cattle breeds to some extent while they have been almost differentiated. In population structure analysis, Japanese Black cattle shared little genetic component (3.5%) with European breeds. This is the first study to determine the extent to which European breeds impact Japanese breeds.

Association of SNPs in AKIRIN2, TTN, EDG1 and MYBPC1 Genes with Growth and Carcass Traits in Qinchuan Cattle

Abstract Growth and carcass traits are the main breeding objectives in beef cattle. The aim of this study was to confirm genetic effects of the c.*188G>A SNP of AKIRIN2, the g.231054C>T SNP of TTN, the g.1471620G>T SNP of EDG1 and the g.70014208A>G SNP of MYBPC1 on growth and carcass traits in Chinese Qinchuan (QC) cattle, as well as to compare the frequencies of the well-characterized alleles of these SNPs among six Chinese cattle populations, three Japanese cattle populations, two European cattle populations and one Korean cattle population. In this study, a total of 665 cattle samples were genotyped using MassARRAY and PCR-RFLP. Association analysis explored effects of four SNPs on growth and carcass traits including body length, wither height, hip height, hip width, rump length, chest depth, chest circumference, back fat thickness, ultrasound longissimus muscle area and ultrasound longissimus muscle depth in QC (P<0.05 to P<0.001). The well-characterized A (c.*188G>A), T (g.231054C>T) and T (g.1471620G>T) alleles in Japanese Black cattle were significantly higher than Chinese cattle breeds; on the contrary, the G allele (g.70014208A>G) was markedly higher in Chinese cattle breeds than other cattle breeds. These results suggest that the four SNPs might be useful as a molecular marker for growth-related traits in Chinese QC cattle.

Genetic variation in Japanese Holstein cattle for EBL development

BackgroundInfection with bovine leukemia virus (BLV), the causative agent for enzootic bovine leukosis (EBL), is increasing in dairy farms of Japan. The tendency of tumor development following BLV infection in certain cow families and bull lines has previously been described. We therefore hypothesized the existence of a genetic component which differentiates cattle susceptibility to the disease.ResultsWe analyzed routinely collected large-scale data including postmortem inspection data, which were combined with pedigree information and epidemiological data of BLV infection. A total of 6,022 postmortem inspection records of Holstein cattle, raised on 226 farms served by a regional abattoir over 10 years from 2004 to 2015, were analyzed for associations between sire information and EBL development. We then identified statistically the relative susceptibility to EBL development for the progeny of specific sires and paternal grandsires (PGSs). The heritability of EBL development was calculated as 0.19. Similarly, proviral loads (PVLs) of progeny from identified sires and PGSs were analyzed, but no significant differences were found.ConclusionsThese observations suggest that because EBL development in our Holstein population is, at least in part, influenced by genetic factors independent of PVL levels, genetic improvement for lower incidence of EBL development in cattle notwithstanding BLV infection is possible.

Genetic characteristics of Korean Jeju Black cattle with high density single nucleotide polymorphisms.

ObjectiveConservation and genetic improvement of cattle breeds require information about genetic diversity and population structure of the cattle. In this study, we investigated the genetic diversity and population structure of the three cattle breeds in the Korean peninsula.MethodsJeju Black, Hanwoo, Holstein cattle in Korea, together with six foreign breeds were examined. Genetic diversity within the cattle breeds was analyzed with minor allele frequency (MAF), observed and expected heterozygosity (HO and HE), inbreeding coefficient (FIS) and past effective population size. Molecular variance and population structure between the nine breeds were analyzed using a model-based clustering method. Genetic distances between breeds were evaluated with Nei’s genetic distance and Weir and Cockerham’s FST.ResultsOur results revealed that Jeju Black cattle had lowest level of heterozygosity (HE = 0.21) among the studied taurine breeds, and an average MAF of 0.16. The level of inbreeding was −0.076 for Jeju Black, while −0.018 to −0.118 for the other breeds. Principle component analysis and neighbor-joining tree showed a clear separation of Jeju Black cattle from other local (Hanwoo and Japanese cattle) and taurine/indicine cattle breeds in evolutionary process, and a distinct pattern of admixture of Jeju Black cattle having no clustering with other studied populations. The FST value between Jeju Black cattle and Hanwoo was 0.106, which was lowest across the pair of breeds ranging from 0.161 to 0.274, indicating some degree of genetic closeness of Jeju Black cattle with Hanwoo. The past effective population size of Jeju Black cattle was very small, i.e. 38 in 13 generation ago, whereas 209 for Hanwoo.ConclusionThis study indicates genetic uniqueness of Jeju Black cattle. However, a small effective population size of Jeju Black cattle indicates the requirement for an implementation of a sustainable breeding policy to increase the population for genetic improvement and future conservation.

Prussian blue in salt blocks decreases radiocesium activity concentration in milk from dairy cattle fed a diet contaminated by the Fukushima nuclear accident

In Japan, the radiocesium activity concentration in milk must be less than 50 Bq/kg-fresh to meet shipping standards, and the radiocesium concentration of the diet fed to dairy cattle must be less than 500 Bq/kg-dry. After the Fukushima nuclear accident in 2011, we conducted two experiments to investigate whether Prussian blue (PB) could suppress the radiocesium (134Cs + 137Cs) activity concentration in Japanese cattle’ milk. In experiment 1, four cattle were fed a diet with a radiocesium activity concentration of 175 Bq/kg-dry, with or without PB supplementation. The PB intake ranged from 0 to 3.0 g/day, and the average radiocesium intake was 3.42 kBq/day in all treatments. The radiocesium activity concentration in milk decreased from 16.4 to 8.6 Bq/kg-fresh, and the transfer coefficient of radiocesium from diet to milk (Fm) decreased from 4.77 × 10−3 to 2.61 × 10−3 with increased PB intake. In experiment 2, three cattle were fed another diet including a radiocesium activity concentration of 927 Bq/kg-dry of with or without PB supplementation. The PB intake ranged from 0 to 18.9 g/day, and the average radiocesium intake was 15.2 kBq/day in all treatments. The milk's radiocesium activity concentration decreased from 24.3 to 4.2 Bq/kg-fresh, and the Fm decreased from 1.68 × 10−3 to 0.28 × 10−3 with increased PB intake. Our results suggest that both the radiocesium activity concentration in milk and Fm can be reduced by PB, and that Fm is affected by diet. We recommend cattle should be fed absorbents such as PB to minimize the risk of milk radiocesium activity concentration exceeding 50 Bq/kg-fresh even if the diet has a radiocesium activity concentration of less than 500 Bq/kg-dry.

Molecular Detection and Subtyping of Blastocystis Detected in Wild Boars (Sus scrofa) in South Korea.

Blastocystis is a genus of parasitic protozoans that live in humans, mammals, and birds and which has been widely studied due to its low host specificity. Limited data are available, however, regarding its presence in wildlife, particularly in South Korea. Contact between wild boars (Sus scrofa) and livestock or humans has steadily increased as wild boars venture down from the mountains to farms and residential areas. In this study, we examined the status and subtypes (STs) of Blastocystis in wild boars in South Korea and confirmed its zoonotic potential. From March 2016 to November 2018, we collected 433 fecal samples throughout the country from trapped or road-killed wild boars. The 18S rRNA gene was used for molecular identification and subtyping and the proportion of PCR-positive samples was 10.4%. We then assessed positive samples for associations with sex, region, and seasonal infection; however, no statistical significance was observed for any variable other than season. Phylogenetic analyses revealed that all sequences belonged to subtype 5 and had 99.5-99.9% identity with sequences obtained from Japanese cattle (Bos taurus) and 97.1% identity with sequences obtained from Chinese. Subtype 5 has been implicated in zoonoses, indicating that Korean wild boars could transmit Blastocystis to humans and other livestock. Our results, in accordance with the One Health concept, strongly support continued interest and efforts by public health and disease control organizations toward transmission prevention.

Detection of Spalangia (Hymenoptera: Pteromalidae) attacking Stomoxys calcitrans (Diptera: Muscidae) and their potential significance in the Japanese cattle industry

Four species of Spalangia Latreille (Hymenoptera: Pteromalidae), S. cameroni Perkins, S. endius Walker, S. nigroaenea Curtis, and S. nigra Latreille, were collected from dairy farms located in Fukuoka, Japan. The former three species were reared from pupae of Stomoxys calcitrans (Linnaeus) (Diptera: Muscidae) and the latter was reared from pupae of Chrysomya pinguis (Walker) (Diptera: Calliphoridae). This is a new distributional record for S. cameroni from Japan. Chrysomya pinguis is newly recorded as the host insect of S. nigra. A key to Japanese species of Spalangia was provided for further biological studies. Spalangia species have been known as biological control agents against St. calcitrans in Europe and North America. Thus, the detection of these parasitoids provides opportunities for biological control as an additional control measure against St. calcitrans in Japan.

Molecular characterization of a new Trypanosoma (Megatrypanum) theileri isolate supports the two main phylogenetic lineages of this species in Japanese cattle.

Trypanosoma (Megatrypanum) theileri is a cosmopolitan, usually non-pathogenic, trypanosome of cattle transmitted by blood-sucking arthropods, mainly tabanid flies. Several T. theileri strains isolated from domestic and wild ruminants via co-culturing with mammalian feeder cells or blood cells have been characterized morphologically and genetically. Here, we cultured a new trypanosome isolate from a Holstein cow in Hokkaido, Japan, and performed morphological and molecular characterization studies. The new isolate (Obihiro strain) was co-cultivated with Madin-Darby bovine kidney (MDBK) cells in GIT medium supplemented with 10% fetal bovine serum. Trypomastigotes and epimastigotes, but not intracellular parasites, were identified in the culture. Analysis of the V7-V8 region of 18S rRNA sequences showed that the Obihiro strain is positioned within the subgenus Megatrypanum. A dendrogram based on whole internal transcribed spacer rDNA sequence showed that the Obihiro strain clustered in the lineage TthII together with the Japanese isolates of T. theileri, Esashi 9, and Esashi 12, and isolates from Zambia and the USA. T. theileri of the KM strain and a T. theileri-like trypanosome isolated from deer (TSD1 strain) clustered in the lineage TthI, separate from the Obihiro strain. Based on a partial cathepsin L-like protein gene analysis, the Obihiro strain clustered with isolates of the TthIIF genotype, which includes T. theileri from Vietnam, Sri Lanka, and Brazil. Our analyses of the T. theileri Obihiro strain provide relevant insights into its genetic diversity in Japanese cattle and corroborate the host specificity of cattle and deer trypanosomes of the subgenus Megatrypanum.

Development of a structural growth curve model that considers the causal effect of initial phenotypes

BackgroundGrowth curves have been widely used in genetic analyses to gain insights into the growth characteristics of both animals and plants. However, several questions remain unanswered, including how the initial phenotypes affect growth and what is the duration of any such impact. For beef cattle production in Japan, calves are procured from farms that specialize in reproduction and then moved to other farms where they are fattened to achieve their market/purchase value. However, the causal effect of growth, while calves are on the reproductive farms, on their growth during fattening remains unclear. To investigate this, we developed a model that combines a structural equation with a growth curve model. The causal effect was modeled with B-splines, which allows inference of the effect as a curve. We fitted the proposed structural growth curve model to repeated measures of body weight from a Japanese beef cattle population (n = 3831) to estimate the curve of the causal effect of the calves’ initial weight on their trajectory of growth when they are on fattening farms.ResultsMaternal and reproduction farm effects explained 26% of the phenotypic variance of initial weight at fattening farms. The structural growth curve model was fitted to remove the effects of these factors in growth curve analysis at fattening farms. The estimated curve of causal effects remained at approximately 0.8 for 200 d after the calves entered the fattening farms, which means that 64% of the phenotypic variance was explained by the initial weight. Then, the effect decreased linearly and disappeared approximately 620 d after entering the fattening farms, which corresponded to an average age of 871.5 d.ConclusionsThe proposed model is expected to provide more accurate estimates of genetic values for growth patterns because the confounding causal factors such as maternal and reproduction farm effects are removed. Moreover, examination of the inferred curve of the causal effect enabled us to estimate the effect of a calf’s initial weight at arbitrary times during growth, which could provide suitable information for decision-making when shifting the time of slaughter, building models for genetic evaluation, and selecting calves for market.

Planning Large-scale Feed Production for Japanese Cattle Farming under the Condition of Cropland Dispersion

Planning Large-scale Feed Production for Japanese Cattle Farming under the Condition of Cropland Dispersion

Effects of surrounding fluid on motility of hyperactivated bovine sperm

Mammalian spermatozoa in organisms with internal fertilization are required to swim in the cervical and oviductal mucus, whose rheological properties differ substantially from those of water. Moreover, on the way to the oviduct, a change in sperm motility called hyperactivation may occur. In the present study, we focused on the motion characteristics of hyperactivated bovine sperm and investigated the effect of the surrounding fluid on motility. We prepared two kinds of polyacrylamide with high-viscosity non-Newtonian fluid properties, similar to the actual cervical and oviductal mucus. Using semen from Japanese cattle, we evaluated curvilinear velocity (VCL), straight-line velocity (VSL), and average path velocity (VAP). Additionally, we estimated linearity (LIN), straightness (STR), and wobble (WOB) as sperm motility parameters for several surrounding fluids. We successfully induced hyperactivation of bovine sperm in high-viscosity non-Newtonian fluid. Hyperactivation resulted in an increase in VCL and a decrease in VSL. In the high-viscosity non-Newtonian fluid, the hyperactivated sperm moved in a zig-zag pattern with regularity, different from the movement observed in a diluted solution. The increase in WOB in the non-Newtonian fluid suggests that hyperactivated sperm efficiently progress along the groove that exists on the oviductal mucus wall. These results improve our understanding of the motility of bovine sperm when they undergo hyperactivation in the actual cervical and oviductal mucus.

Time Series Analysis for Shortened Labor Mean Interval of Dairy Cattle with the Data of BCS, RFS, Weight, Amount of Milk and Outlook

MTime series analysis for shortened labor mean interval of dairy cattle with the data of Body Condition Score (BCS), Rumen Fill Score (RFS), Weight, Amount of Milk and Outlook is conducted. Method for shortened the labor mean internal of Japanese dairy cattle based on time-series analysis with the data of visual index of BCS, RFS, Weight, Amount of Milk and Outlook is proposed. In order to shortened the labor mean interval of dairy cattle is the purpose of the research. Through the experiments with 17 Japanese dairy cattle of the 17 Japanese anestrus Holstein dairy cattle, it is found that the combination of weight, BCS and amount of milk is a good indicator for identification of productive cattle. Therefore, the cattle which need hormone treatments can be identified.

First detection and molecular identification of Entamoeba bovis from Japanese cattle.

Thus far, Entamoeba species have been classified based on morphology such as the number of nuclei in mature cysts and their hosts. Using recently developed molecular tools, ruminant Entamoeba spp. are currently classified into four species/genotypes: E. bovis and Entamoeba ribosomal lineages (RL) 1, 2, and 4. However, the distribution or pathogenicity of ruminant Entamoeba has not been well documented. In the present study, we examined a total of 25 fecal and seven environmental samples collected from six farms in Japan from 2016 to 2017 by the floatation method and PCR and sequencing analyses. Consequently, we detected Entamoeba cysts in 18 of 25 cattle samples and four of the seven environmental samples, including soil and drinking water, by microscopic examinations. In sequential examinations, Entamoeba-positive cattle were found to shed cysts without any clinical symptoms for more than 8months. By PCR for molecular identification, isolates in ten cattle and one soil sample were successfully sequenced and formed a cluster of E. bovis, which was separated from those of other Entamoeba species/genotypes such as RL1-4 in phylogenetic analysis. To our knowledge, this is the first report about E. bovis in Japan, and our results may implicate that E. bovis is not pathogenic.

Partial characterization of phylogeny, ecology and function of the fibrolytic bacterium Ruminococcus flavefaciensOS14, newly isolated from the rumen of swamp buffalo.

The fibrolytic rumen bacterium Ruminococcus flavefaciensOS14 was isolated from swamp buffalo and its phylogenetic, ecological and digestive properties were partially characterized. Isolates from rumen contents of four swamp buffalo were screened for fibrolytic bacteria; one of the 40 isolates showed a distinctive feature of solubilizing cellulose powder in liquid culture and was identified as R. flavefaciens based on its 16S ribosomal DNA sequence. This isolate, OS14, was employed for detection and digestion studies, for which a quantitative PCR assay was developed and defined cultures were tested with representative forages in Thailand. OS14 was phylogenetically distant from other isolated and uncultured R. flavefaciens and showed limited distribution among Thai ruminants but was absent in Japanese cattle. OS14 digested rice straw and other tropical forage to a greater extent than the type strain C94 of R. flavefaciens. OS14 produced more lactate than C94, and digested para grass to produce propionate more extensively in co-culture with lactate-utilizing Selenomonas ruminantium S137 than a co-culture of C94 with S137. These results indicate that phylogenetically distinct OS14 could digest Thai local forage more efficiently than the type strain, possibly forming a symbiotic cross-feeding relationship with lactate-utilizing bacteria. This strain might be useful for future animal and other industrial applications.