Relationships between the first ovulation postpartum and polymorphism in genes relating to function of immunity, metabolism and reproduction in high-producing dairy cows

The decrease in fertility and conception rates of high-producing dairy cows is one of the major negative impacts for today's producers. The recovery of ovarian activity postpartum is affected by the status of immunity, metabolism and reproduction and plays a critical role in subsequent fertility after parturition in the cow. In the present study we investigated the relationships between polymorphisms in genes relating to the above functions and the first postpartum ovulation as a marker of the recovery of ovarian function in the cow. In immune function related-factors, the occurrence of first postpartum ovulation within 3 weeks in the C/C genotypes of tumor necrosis factor α (TNFα) exon (55.4%) and the A/G genotypes of TNFα promoter (55.4%) was significantly higher than that in T/T genotypes of TNFα exon (14.3%) and A/A genotypes of TNFα promoter (14.3%). Moreover, anovulatory cows with the T/T genotype of TNFα exon and the A/A genotype of TNFα promoter tended to have a prolonged days open compared with those of the other genotypes of TNFα polymorphisms. In metabolic function-related factors, ovulatory and anovulatory cows had a different distribution for alleles of the growth hormone receptor, but there were no significant differences in genotype and allele frequency of insulin-like growth factor-I polymorphism. No significant relationships were found between ovarian function after parturition and polymorphisms for reproduction-related genes. In conclusion, polymorphisms of TNFα gene both in exon and promoter regions have a strong association with the early first ovulation within 3 weeks after parturition in the high-producing dairy cow. Taken together, polymorphisms of TNFα gene could be strongly related to early first ovulation after parturition, thus being an effective tool of selection for improving reproductive performance in the high-producing dairy cow.     

Development of discrimination markers between Japanese domestic and imported beef

In the meat industry, correct labeling of beef origins or breed is required to assure quality and safety. This paper describes the development of discrimination markers between Japanese domestic and imported beef from the United States (US) and Australia (AUS) based on a bovine 50K single nucleotide polymorphism (SNP) array using a total of 110 samples: Japanese Black (n = 50), Japanese Holstein (n = 50) and US cattle (n = 10). Genotyping information revealed 1081 SNPs as candidate markers that were polymorphic only in US cattle. The genotyping results by PCR – restriction length polymorphism in Japanese Black (n = 300) and Holstein cattle (n = 146) revealed that 11 SNPs had alleles specific to US cattle. Their allelic frequencies in US cattle (n = 108) ranged from 0.097 to 0.250 with an average of 0.178 and the combined identification probability of US cattle was 0.987. In addition, we also verified the applicability of these US‐specific markers to AUS cattle. Their allelic frequencies in AUS cattle (n = 280) ranged from 0.063 to 0.224 with an average of 0.137 and the combined identification probability of AUS cattle was 0.963. In conclusion, a set of these markers could be useful for discriminating between Japanese domestic and imported beef and would contribute to identify origins and prevent falsified labeling of beef.     

Identification of tamaraw (Bubalus mindorensis) from natural habitat‐derived fecal samples by PCR‐RFLP analysis of cytochrome b gene

Fecal DNA analysis is a useful tool for the investigation of endangered species. Tamaraw (Bubalus mindorensis) is endemic to the Philippine island of Mindoro but knowledge of its genetic and ecological information is limited. In this study, we developed a species identification method for tamaraw by fecal DNA analysis. Eighteen feces presumed to be from tamaraw were collected in Mount Iglit‐Baco National Park and species‐known feces from domestic buffaloes and cattle were obtained from a farm. Additionally, one species‐unknown fecal sample was obtained in Mount Aruyan Preserve, where the sighting of tamaraw has not been reported in recent years. Based on DNA sequence data previously reported, the genus Bubalus‐ and tamaraw‐specific primers for PCR of cytochrome b gene were newly designed. The Bubalus‐specific primer yielded a 976 bp fragment of cytochrome b for all fecal samples from tamaraw and domestic buffaloes, but not for cattle, whereas the tamaraw‐specific primer yielded a 582 bp fragment for all tamaraw fecal samples and for one of the four domestic buffalo samples. PCR‐RFLP (restriction fragment length polymorphism) analysis of the 976 bp PCR fragment with AvrII or BsaXI provided distinct differences between tamaraw and domestic buffalo. PCR‐RFLP analysis also showed that the species‐unknown sample obtained in Mount Aruyan Preserve, originates from tamaraw.     

Molecular diversity of bacterial chitinases in arable soils and the effects of environmental factors on the chitinolytic bacterial community

The molecular diversity of bacterial chitinases in the bulk soils of arable land was investigated using culture-independent methods. The results demonstrate that bacterial chitinases in arable soils are highly diverse and comprise unique groups when their sequences were compared to those in public databases. The diversity of bacterial chitinases in arable soil was further evaluated using conventional phylogenetic analysis, the UniFrac analysis of the phylogenetic data, and the multidimensional scaling (MDS) analysis of T-RFLP profiles to elucidate the relationship between the diversity of bacterial chitinases and soil characteristics. These analyses indicate that environmental factors such as soil type and pH are responsible for shaping the composition of bacterial chitinases.     

A new taxonomic treatment for some wild relatives of mungbean (<Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilcz.) based on their molecular phylogenetic relationships and morphological variations

Mungbean (Vigna radiata (L.) Wilcz.) is an important leguminous crop cultivated mainly in Asia. Its wild relatives are considered useful genetic resources for mungbean breeding. However, the taxonomic history of mungbean and its wild relatives is complicated and some confusion is still present in recent publications. In this study, we examined the rDNA-ITS sequences and morphological characteristics of 83 gene bank accessions closely related to mungbean. As a result, we classified the 83 accessions into five species and one unclassified accession. The proper taxon name for each species was determined (Vigna grandiflora (Prain) Tateishi et Maxted, Vigna mungo (L.) Hepper, V. radiata, Vigna subramaniana (Babu ex Raizada) Raizada, and Vigna trinervia (Heyne ex Wight et Arn.) Tateishi et Maxted) based on a review of their taxonomic history and morphological comparisons between gene bank accessions and type specimens. A new taxonomic treatment is proposed and a morphological key has been prepared. In this treatment, Vigna radiata (L.) Wilcz. var. setulosa (Dalz.) Ohwi et Ohashi is treated as a synonym of Vigna radiata (L.) Wilcz. var. sublobata (Roxb.) Verdc., and Vigna hainiana Babu, Gopinathan et Sharma is a synonym of V. subramaniana. Accession ‘NI1135’ was revealed to be most closely related to, but is considerably differentiated from, mungbean (V. radiata) based on its rDNA-ITS sequences. It also has distinguishing morphological characters. Plants with similar morphologies and DNA profiles might be distributed in the Indian Himalaya. However, since only one accession is available at present, the taxonomic classification of ‘NI1135’ needs to be reconsidered in the future.     

Detection of Babesia microti-like parasite in filter paper-absorbed blood of wild rodents

The first case of human babesiosis was reported in Japan. The epidemiology of this disease in Japanese nature remains unclear. In this study, 97 common field mice captured in Hokkaido, Japan, were examined. Blood specimens absorbed onto filter papers were eluted and tested by nested PCR using specific primers for the B. microti nuclear small subunit rRNA genome. Twenty-three percent (11/47) of Apodemus speciosus and four percent (2/50) of Clethrionomys rufocanus were positive. The 159-bp primary sequences of PCR products tested exhibited 97.5% and 96.8% homology with those of the human isolate in Japan and of U.S. strains of B. microti, respectively.