The effects of short-term nutritional stimulus before and after the luteolysis on metabolic status, reproductive hormones and ovarian activity in goats

The effect of short-term nutritional supplementation on hormonal and ovarian dynamics was studied in goats. Cycling Shiba goats were divided randomly into maintenance (group M, n=4) and high-energy (group H, n=4) groups. After the detection of the ovulation (Day 0, 1(st) ovulation), group H received a high-energy diet providing 2.5 times of the maintenance energy requirement for 7 days from Day 7 to Day 13 and were administered 2 mg of prostaglandin F(2alpha) (PGF(2alpha)) on Day 10 to induce luteal regression followed by the follicular phase. Follicular and luteal dynamics were monitored using ultrasonography daily or every other day, and blood samples were collected daily from Day 0 to the third ovulation (3(rd) ovulation) following the second ovulation (2(nd) ovulation) induced by PGF(2alpha) administration. Blood samples were also collected at 10-min intervals for 6 h on Day 9 and Day 11 for analysis of pulsatile LH secretion. The mean concentrations of glucose and insulin were significantly (P<0.05) higher in group H than in group M on Days 8, 9, 12, 13 and Days 8, 9 and 10, respectively. For both the 2(nd) and 3(rd) ovulations, no significant difference was detected in ovulation rate between groups M and H. On the other hand, the interpeak interval for wave-like patterns of FSH in group H was significantly (P<0.05) shorter than in group M during the period between the 1(st) and 2(nd) ovulations (4.3 +/- 0.3 vs. 6.5 +/- 1.5 days). The mean LH pulse frequency in group H was significantly (P<0.05) greater than in group M on Day 11 (4.5 +/- 0.6 vs. 3.3 +/- 0.5 pulses/6 h). The present study clearly demonstrated that short-term (7 days) nutritional supplementation promoted pulsatile LH and wave-like FSH secretions in cycling goats. However, no significant increase in ovarian performance was found under such endocrine and metabolic conditions.     

High level expression of Prop-1 gene in gonadotropic cell lines

Prop-1 acts as an upstream regulator for the Pit-1 gene to induce development of Pit-1 lineage pituitary cell lines, GH-, PRL-, and TSH-producing cells, in the early stage of pituitary organogenesis. Furthermore, Prop-1 is presumed to be involved in the function of FSH/LH-producing cells, gonadotropes, since the defective Prop-1 gene shows hypogonadism. Recently, we reported evidence that Prop-1 directly regulates expression of the porcine FSHbeta gene, thus providing a novel advance in understanding the function of Prop-1 in FSH/LH production and hypogonadism. This study was intended to demonstrate the expressions of Prop-1 gene in pituitary tumor-derived cell lines. RT-PCR analyses were conducted of Pit-1, glycoprotein alpha subunit (alphaGSU), GnRH receptor, and cyclophilin A (a ubiquitously expressing gene). We observed expression of the Pit-1 gene in alphaT1-1, TalphaT1, MtT/S, GH3, and TtT/GF cells, expression of the alphaGSU gene in alphaT1-1, alphaT3-1, LbetaT2, LbetaT4, TalphaT1, and GH3 cells, and expression of GnRH receptor gene in alphaT3-1, LbetaT2, LbetaT4, and GH3 cells, respectively. These expression profiles were in accord with their cell lineages, with only a few exceptions. To accurately measure the expression level of the Prop-1 gene, a quantitative analysis was performed using the real-time PCR method. This analysis demonstrated that the LbetaT2 and LbetaT4 gonadotrope cell lines, which express the FSHbeta gene, express the Prop-1 gene. Taken together with our previous observation that Prop-1 is present in the adult porcine pituitary gonadotropes, Prop-1 might also be involved in development of gonadotropes and hormone production.     

Artificial Infection of Rosellinia necatrix with Purified Viral Particles of a Member of the Genus Mycoreovirus Reveals Its Uneven Distribution in Single Colonies

ABSTRACT Rosellinia necatrix mycoreovirus 3 (W370) (RnMYRV-3/W370, described as RnMYRV-3 in this paper), a member of the newly established genus Mycoreovirus within the family Reoviridae, is the hypovirulence factor of the white root rot fungus, Rosellinia necatrix. Two virus-free fungal isolates (W37 and W97) that were somatically incompatible with the virus-harboring field isolate (W370) were transfected with purified RnMYRV-3 particles. Virus infection was confirmed by electrophoresis and northern hybridization of viral double-stranded RNA. RnMYRV-3 was transmissible from transfected strains to their respective, virus-free counterparts via hyphal anastomosis. Virus-transfected strains produced smaller lesions on apple fruits than did their virus-free counterparts. Virus-cured strains were indistinguishable from wild-type strains in culture morphology and displayed approximately the same virulence level on apples. Virus-transfected strains had "mosaic" colony portions consisting of thin, fast-growing and dense, slow-growing mycelia, and grew more slowly as a whole than their virus-free, parental strains. The level of virus accumulation varied among virus-transfected subcultures and within its single colonies. Virus-transfected strains were occasionally cured, as was W370. Such a phenomenon may be ascribed to uneven viral distribution in single colonies and the difficulty in viral transmission to virus-free hyphae.     

A longitudinal study to evaluate the diagnostic potential of a direct faecal quantitative PCR test for Johne's disease in sheep

A longitudinal study was carried out to evaluate the diagnostic potential of the previously developed direct faecal real-time quantitative PCR (QPCR) assay (Kawaji et al., 2007) for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) infected sheep. Of the 58 sheep, 38 were orally inoculated with MAP, while 20 controls were maintained separately from the infected group throughout the trial. All animals were tested by QPCR, faecal culture and serum ELISA pre-inoculation and at 4, 8 and 13 months post-inoculation, and were necropsied at 13 months post-inoculation. Eighteen out of 38 inoculated sheep were detected by QPCR to be shedding MAP in faeces at 4 months post-inoculation, while only one sheep was positive in faecal culture at this time point. At 8 months post-inoculation, MAP DNA was detected in faeces of all inoculated sheep by QPCR, while MAP organisms were isolated from only 34% of the inoculated animals by faecal culture. The QPCR results for faecal samples that were collected at necropsy demonstrated that faecal QPCR was more sensitive than culture of intestinal tissues for MAP. The QPCR assay was confirmed to be a sensitive and specific ante-mortem diagnostic test for MAP in sheep, circumventing faecal culture which is a less sensitive and highly time consuming test. Quantification of MAP DNA in faeces by QPCR may provide immediate information to estimate the stage of the infection as well as the risk of transmission from infected animals.     

The draft genome of Ciona intestinalis: insights into chordate and vertebrate origins

The first chordates appear in the fossil record at the time of the Cambrian explosion, nearly 550 million years ago. The modern ascidian tadpole represents a plausible approximation to these ancestral chordates. To illuminate the origins of chordate and vertebrates, we generated a draft of the protein-coding portion of the genome of the most studied ascidian, Ciona intestinalis. The Ciona genome contains approximately 16,000 protein-coding genes, similar to the number in other invertebrates, but only half that found in vertebrates. Vertebrate gene families are typically found in simplified form in Ciona, suggesting that ascidians contain the basic ancestral complement of genes involved in cell signaling and development. The ascidian genome has also acquired a number of lineage-specific innovations, including a group of genes engaged in cellulose metabolism that are related to those in bacteria and fungi.     

Open Heart Surgery with Deep Hypothermia and Cardiopulmonary Bypass in Small and Toy Dogs

Objective: To evaluate open heart surgery with deep surface‐induced hypothermia (sHT) and low‐flow cardiopulmonary bypass (CPB) in small and toy‐breed dogs. Study Design: Case series. Animals: Small breed dogs (n=8) weighing <5.5 kg with naturally occurring cardiac disease. Methods: Deep sHT under isoflurane anesthesia and low‐flow rate CPB with a small‐volume prime circuit were used. Ventricular septal defect was closed directly in 2 dogs and severe mitral regurgitation was corrected with mitral valvuloplasty (MVP) in 5 dogs and mitral valve replacement in 1 dog. Results: All dogs survived surgery; 1 dog died 6 days and 1 died 2 months after MVP. The other 6 dogs lived (mean follow‐up, 32.8 months; range, 12–65 months). Mean body weight at surgery was 3.6 kg (range, 2–5.3 kg). Mean lowest esophageal temperature was 21.4°C (range, 19.8–23.8°C). Mean lowest pump flow volume was 29.2 mL/kg/min (range, 9.4–57.7 mL/kg/min) during aortic cross‐clamping (mean, 53.5 minutes; range, 25–79 minutes). Mean hematocrit before CPB was 38.6% (range, 33–47%) and 20.3% (range, 13–24%) during CPB with a small circuit priming volume of 225–260 mL. Conclusion: Deep sHT with low‐flow rate CPB may be used for open heart surgery in small dogs weighing <5.5 kg. Clinical Relevance: Open heart surgery for selected congenital defects and acquired defects in small and toy‐breed dogs may be successfully performed using deep sHT and CPB.     

Histopathology of red crown rot of soybean

Histopathological analyses of red crown rot (RCR) of soybean were performed using field-grown and indoor-grown soybean plants. These analyses revealed that soybean plants formed a periderm to prevent pathogen invasion. When a break formed in the periderm, an additional periderm newly differentiated into the inner cells and prevented pathogen invasion. If the pathogen reached the intravascular tissues, some xylem parenchyma cells died, and the vessels were occluded by gum. Periderm formation and vessel occlusion were observed not only in slightly diseased soybean plants, but also in seriously diseased plants, indicating that both reactions function as part of the defense response against RCR.