Enhanced Growth and nodule occupancy of red kidney bean and soybean inoculated with soil aggregate-based inoculant

Volcanic ash soil, which is widely distributed in Japan, contains a large amount of well-structured soil aggregates. By using these aggregates as carrier materials, we prepared (brady)rhizobial inoculants for red kidney bean (Phaseolus vulgaris) and soybean (Glycine max). Autoclaved soil aggregates were inoculated with Rhizobium tropici CIATS99R or Bradyrhizobium japonicum USDA110R, incubated for 15 or 21 d at 30°C, slowly air-dried at 20°C to prepare the aggregate-based inoculants, and stored at various temperatures. The populations of CIATS99R and USDA110R in the aggregate-based inoculants were maintained during several months of storage at 20°C. When the aggregate-based inoculants were mixed with soil, CIATS99R and USDA110R cells showed a remarkably improved survival in soils compared with those mixed with soil without carrier material. The effect of the aggregate-based inoculants on the growth of red kidney bean and soybean was examined in pot experiments. By placing a small amount of the inoculant just beneath the seeds at the time of sowing, plant growth was significantly enhanced compared with the use of traditional peat-based inoculant. In addition, nodule formation on the upper part of soybean roots and nodule occupancy by the inoculated strain were remarkably enhanced by the aggregate-based inoculant. It is suggested that soil aggregates might be suitable carrier materials for preparing cheap and effective (brady)rhizobial inoculants.     

Myb类基因Ibmyb在甘薯不同品种中的表达

研究Ibmyb基因在不同甘薯品种中的表达情况。目前Soedarjo等已从甘薯紫色块根的cDNA文库中克隆出了一条全长1243bp的cDNA序列,经测序及同源性比较,推测该基因为花青苷Myb类转录因子基因,命名为Ibmyb。本研究根据Ibmyb基因cDNA序列设计了2对引物,分别对紫叶的‘#13’和紫色块根的‘Benihikari’的叶和块根进行了RT-PCR分析;以Ibmyb cDNA为探针对这两个品种进行了Southern杂交分析。结果表明该基因在这两个甘薯品种的叶和块根中都表达,在甘薯基因组中具有多个拷贝且品种间不存在限制性片段长度多态性。表明Ibmyb可能是一种在甘薯中普遍存在的蛋白。     

Adjuvant effects of sugar cane extracts (SCE) in chickens

The effects of sugar cane extracts (SCE) on immune responses in chickens were studied. Two- or 10-month-old chickens orally administered SCE (500 mg/kg/day), for 3 consecutive days before immunized with sheep red blood cells, Brucella abortus and Salmonella Enteritidis organisms, showed significantly increased and prolonged antibody responses to these antigens, compared to control chickens without SCE. Furthermore, chickens orally administered SCE also revealed enhanced delayed type hypersensitivity responses to human gamma globulin. These results indicated that SCE has immunostimulating and adjuvant effects in chickens.     

ELISAs based on rNcGRA7 and rNcSAG1 antigens as an indicator of Neospora caninum activation

Bovine abortion caused by the Apicomplexa parasite Neospora caninum is a major economical problem in the livestock industry worldwide. However, the relationship between N. caninum infection and abortion is still unknown. Our study aimed to elucidate the relationship between parasite-specific antibody responses, parasite stages and abortion. In experimentally infected cattle, anti-NcGRA7 IgG1 antibody was predominantly detected during the acute infection stage, while the production of anti-NcSAG1 IgG1 antibody was observed during both acute and chronic stages. Furthermore, levels of anti-NcGRA7 IgG2 antibody were lower than those of anti-NcSAG1 IgG2 antibody. When tested on cattle with Neospora-associated abortion, positive rates of the anti-NcGRA7 IgG2 antibody were significantly lower than those of the anti-NcSAG1 antibody, although there was no difference in IgG1 antibody-positive rates between the two antigens. In addition, anti-NcGRA7 IgG2 antibodies were not detected in cattle for more than 30days after abortion. Our results suggested that anti-NcGRA7 and anti-NcSAG1 antibodies are suitable indicators for the activation stage of N. caninum infection and broad detection of the infection, respectively. In conclusion, the use of recombinant NcGRA7 and NcSAG1-based ELISAs will be useful for evaluating the abortion risk associated with N. caninum infection.     

Physiological changes induced by salt intake in female Spontaneously Diabetic Torii‐Leprfa (SDT fatty) rat,a novel obese type 2 diabetic model

Salt plays an important role in the control of blood pressure in obesity and diabetes mellitus. In this study, we investigated physiological changes such as blood pressure and renal function in salt‐loaded female Spontaneously Diabetic Torii‐Lepr^fa (SDT fatty) rats. SDT fatty rats were given 1% NaCl in drinking water for 14 weeks, from 4 to 18 weeks of age. Significant salt‐sensitive hypertension was observed in the salt‐loaded SDT fatty rats. Moreover, the salt‐loaded rats showed a decrease of creatinine clearance and deterioration on pathological renal findings, including glomerulosclerosis and tubular and interstitial lesions. Female SDT fatty rat is a useful model for investigating the mechanisms of high salt sensitivity in obesity and diabetes mellitus.     

Utility of progesterone receptor-ires-Cre to generate conditional knockout mice for uterine study

In mice, the conditional knockout strategy using the Cre-loxP system is useful for various types of research. The Cre mouse line with progesterone receptor promoter (Pgr^Cre) has been widely used to produce specific uterine gene-deficient mice, but in the Cre line, endogenous Pgr gene is replaced by Cre recombinase gene, which makes the breeding of homozygous mice (Pgr^Cre/Cre) difficult because they are infertile. Yang et al. (2013, https://10.1016/j.cell.2013.04.017 ) reported the generation of another Pgr^iresCre mouse line that still has endogenous Pgr gene, and they inserted Cre recombinase downstream of the Pgr gene via an internal ribosome entry site (IRES). It is possible that this new Pgr^iresCre line would be useful for uterine research as the mice can be bred as homozygotes (Pgr^{iresCre/iresCre}). Herein, we confirmed the Pgr^iresCre mice effectively directed recombination in the female reproductive tract and was capable of genetic alteration in the endometrium that enables the studies of its uterine function. Our findings demonstrate that the new Pgr^iresCre mouse line is also useful for the generation of uterine-specific knockout mice. The findings using Pgr^iresCre mouse will contribute to the understanding of reproductive systems and diseases in humans and domestic animals.     

Blood pressure characteristics of female spontaneously diabetic Torii-Lepr(fa) rats

Blood pressure in female SDT-fa/fa rats was periodically investigated at ages 8, 16, and 24 weeks. Furthermore, an insulin therapy was performed for 5 weeks in the female rats at age 11 weeks, and the change of blood pressure was examined. In addition to obesity, hyperglycemia, hyperinsulinemia, and hyperlipidemia, hyperleptinemia and increased urinary angiotensinogen level were observed during the experimental period. Blood pressure was elevated at ages 8 and 16 weeks, but that at 24 weeks was comparable to that in Sprague-Dawley (SD) rats. Heart rate was decreased from age 8 to 24 weeks. Insulin therapy induced good glycemic control and improvement of hyperlipidemia, but the blood pressure was not reduced. Blood pressure in female SDT-fa/fa rats was elevated temporarily. The blood pressure was not decreased by insulin treatment. SDT-fa/fa rat is a useful model to investigate the relation between diabetes mellitus and hypertension.     

Hypothermic storage of porcine zygotes in serum supplemented with chlorogenic acid

The current study was conducted to investigate the effects of 100% foetal bovine serum (FBS) and 100% porcine follicular fluid (pFF) as a storage medium on the developmental competence of porcine zygotes stored at 25°C for 24 hr. Moreover, we evaluated the additive effects of chlorogenic acid (CGA) in the storage medium. When in vitro‐produced zygotes were stored at 25°C for 24 hr in tubes containing either tissue culture medium (TCM) 199 supplemented with 1 mg/ml bovine serum albumin (BSA), 100% of FBS or 100% of pFF, the rate of blastocyst formation was significantly higher in 100% of FBS than in BSA‐containing TCM 199. When the effects of CGA supplementation in 100% of FBS on the development of zygotes stored at 25°C for 24 hr was evaluated, more zygotes stored with 50 µM CGA developed to blastocysts compared with the other concentrations of CGA. When the formation date and quality of blastocysts derived from zygotes stored in 100% of FBS supplemented with 50 µM CGA were investigated, the highest ratio of blastocysts formation in the storage group appeared 1 day later than in the non‐stored control group. However, a higher proportion of blastocysts with apoptotic nuclei was observed in the stored group as compared to the non‐stored group. In conclusion, 100% of FBS is available for a short storage medium of porcine zygotes. The supplementation of 50 µM CGA into the storage medium improves the rates of blastocyst formation of zygotes after storage, but the quality of embryos from the stored zygotes remains to be improved.