Methane production potential and methanogenic archaeal community structure in tropical irrigated Indian paddy soils

Soil characteristics regulate various belowground microbial processes including methanogenesis and, consequently, affect the structure and function of methanogenic archaeal communities due to change in soil type which in turn influences the CH₄ production potential of soils. Thus, five different soil orders (Alfisol, Entisol, Inceptisol, Podzol and Vertisol) were studied to assess their CH₄ production potential and also the methanogenic archaeal community structure in dryland irrigated Indian paddy soils. Soil incubation experiments revealed CH₄ production to range from 178.4 to 431.2 μg CH₄ g^-1 dws in all soil orders as: Vertisol<Inceptisol<Entisol<Podzol<Alfisol. The numbers of methanogens as quantified using real-time quantitative polymerase chain reaction (qPCR) targeting mcrA genes varied between 0.06 and 72.97 (×10⁶ copies g^-1 dws) and were the highest in Vertisol soil and the least in Alfisol soil. PCR-denaturing gradient gel electrophoresis (DGGE)-based approach targeting 16S rRNA genes revealed diverse methanogenic archaeal communities across all soils. A total of 43 DGGE bands sequenced showed the closely related groups to Methanomicrobiaceae, Methanobacteriaceae, Methanocellales, Methanosarcinaceae, Methanosaetaceae and Crenarchaeota. The composition of methanogenic groups differed among all soils and only the Methanocellales group was common and dominant in all types of soils. The highest diversity of methanogens was found in Inceptisol and Vertisol soils. Methane production potential varied significantly in different soil orders with a positive relationship (p?<?0.05) with methanogens population size, permanganate oxidizable C (POXC) and CO₂ production. The present study suggested that CH₄ production potential of different soils depends on physicochemical properties, methanogenic archaeal community composition and the population size.     

Production of Diabetic Offspring Using Cryopreserved Epididymal Sperm by In Vitro Fertilization and Intrafallopian Insemination Techniques in Transgenic Pigs

Somatic cell nuclear transfer (SCNT) is a useful technique for creating pig strains that model human diseases. However, production of numerous cloned disease model pigs by SCNT for large-scale experiments is impractical due to its complexity and inefficiency. In the present study, we aimed to establish an efficient procedure for proliferating the diabetes model pig carrying the mutant human hepatocyte nuclear factor-1α gene. A founder diabetes transgenic cloned pig was generated by SCNT and treated with insulin to allow for normal growth to maturity, at which point epididymal sperm could be collected for cryopreservation. In vitro fertilization and intrafallopian insemination using the cryopreserved epididymal sperm resulted in diabetes model transgenic offspring. These results suggest that artificial reproductive technology using cryopreserved epididymal sperm could be a practical option for proliferation of genetically modified disease model pigs.     

Estimation Models for the Morbidity of the Horses Infected with Equine Influenza Virus

Estimation formulas for the morbidity of horses infected with equine influenza virus by linear regression, logistic regression and probit transformation were developed, using data from the outbreak at the Sha Tin Racing Track in Hong Kong in 1992. Using these formulas, we estimated the equine influenza virus morbidity rates at training centers belonging to the Japan Racing Association (JRA) in October 1997 and in October 1998. In 1998 JRA started a new vaccination program, and every horse must now be vaccinated twice per year. At that time, the vaccine included two US lineage virus strains, the A/equine/Kentucky/81 strain and the A/equine/La Plata/93 (LP93) strain, against equine type-2 influenza viruses; it did not include any EU lineage virus strains, such as A/equine/Suffolk/89 (SF89). Comparing the geometric mean (GM) values of hemagglutination inhibition (HI) titers between the LP93 strain and the SF89 strain in 1997 and in 1998, they both rose significantly at every age (p<0.05) by Wilcoxon test. Calculations by the simulation models show the morbidity rates for LP93 diminished from 0.439 (linear), 0.423 (logistic) and 0.431 (probit) to 0.276 (linear), 0.265 (logistic) and 0.271 (probit), respectively. On the other hand, the estimated morbidity rates for SF89 diminished only slightly from 0.954 (linear), 0.932 (logistic) and 0.944 (probit) to 0.946 (linear), 0.914 (logistic) and 0.927 (probit), respectively. Our simulation models could estimate the effect of the vaccine on each of the equine virus strains represented by the morbidity of infected horses. Thus, they are useful for vaccine evaluation.     

Immunoadjuvant activity of crude lectin extracted from Momordica charantia seed

The aim of this study is to investigate the immunoadjuvant activity of the crude Momordica charantia lectin (crMCL) extracted from seed using beta-galactosidase (beta-gal) as the model antigen. BALB/c mice were injected intramuscularly with beta-gal alone or beta-gal + crMCL for up to four immunizations at two-week intervals. After administration of 2 doses, the IgG-specific titer to beta-gal was significantly higher in mice in the beta-gal + crMCL group than in that from the animals from the beta-gal alone group, while it was about the same in both groups after 1 dose. Our data suggest that crMCL may help raise antibodies under the prime and boost administration regimen and could be a potent vaccine adjuvant.     

Genome‐wide association studies identified variants for taurine concentration in Japanese Black beef

We performed genome‐wide association studies (GWAS) using the BovineSNP50 array to detect significant single nucleotide polymorphisms (SNPs) that may affect the concentration of 22 free amino acids and three peptides in Japanese Black beef cattle. A total of 574 Japanese Black cattle and 40,657 SNPs from the array were used for this study. Genome‐wide significant SNPs were detected for β‐alanine (three SNPs on chromosomes 22 and 29) and taurine (26 SNPs on chromosome 22). Importantly, the top two SNPs for taurine were highly significant (p = 6.2 × 10^?21), and the frequency of the increase‐concentration allele (Q) for taurine was found to be 0.73. The Q allele frequency of this population was similar to that of the other unrelated Japanese Black cattle, but different from that of the other breeds. In addition, the significant SNPs were not associated with carcass traits or fatty acid compositions. Interestingly, the top three of the four most significant SNPs for taurine were located near solute carrier family 6, member 6 (SLC6A6), which is a membrane transporter for taurine. We also found two associated variants in the 5′‐upstream region of SLC6A6; however, they were less significantly associated than the SNPs from the BovineSNP50 array.