In vitro Developmental Competence of Adult Sheep Oocytes Treated with Roscovitine

The efficiency of in vitro sheep embryo production is still low compared to that observed in vivo and in other species. In this context, meiotic inhibition strategies emerged as a promising alternative to improve this biotechnology. So, this study aimed to evaluate, for the first time, the effects of roscovitine on in vitro maturation of sheep oocytes and their subsequent embryo development. For this, cumulus–oocyte complexes (COCs) were cultured for 6 h in the presence (Rosco) or absence (Control) of 75 μm roscovitine and, subsequently, in vitro matured (IVM) for 18 h with gonadotropins. At 0 (Immature), 6 and 24 h of culture, the nuclear status of oocytes was evaluated by Hoechst staining. Embryo cleavage and blastocyst formation were recorded 30 h after in vitro fertilization and on day 7 of culture, respectively. Blastocyst quality was evaluated by differential staining. At 6 h, the GV rate in the Rosco treatment (93.8%) was similar to that observed in the Immature oocytes (94.9%) and significantly higher compared to Control (41.3%). After IVM for 18 h, a high and similar proportion of oocytes from Rosco (93.6%) and Control (88.4%) reached the MII stage. In both treatments, approximately 70% of oocytes cleaved and 50% of them developed up to blastocyst. The mean percentage of blastocyst cells, embryoblast, trophoblast and pyknosis did also not differ between Control and Rosco. In conclusion, roscovitine, at the studied experimental conditions, was efficient to reversibly inhibit the meiosis of adult sheep oocytes without detrimental effect on development and quality of the in vitro produced embryos.     

Evaluation of an enzyme-linked immunosorbent assay for the detection of Mycoplasma hyopneumoniae antibody in porcine serum

An enzyme-linked immunosorbent assay (ELISA) for detecting antibody to Mycoplasma hyopneumoniae in porcine serum is described. The results are presented as an ELISA ratio, calculated by dividing the absorbance of the test sample by the mean absorbance of control negative sera. In known infected pigs, the ELISA ratio was highest when the serum concentration applied to the ELISA plate was diluted 1 in 20 in PBS - Tween. Mean ELISA ratios ranged from 1.2 +/- 0.3 for pigs without porcine enzootic pneumonia (PEP) lesions to 5.5 +/- 1.5 for pigs observed with a PEP lesion reacting positively with immunofluorescent histopathology. Pigs observed with typical PEP lesions at slaughter, but not confirmed by immunofluorescent histopathology had a mean ELISA ratio of 4.9 +/- 1.7. The ELISA was highly sensitive (95.6%) and specific (98.8%) when pig sera from commercial piggeries of known M hyopneumoniae infection status were assessed. No cross-reactivity with serum from a pig hyperimmunised with killed M flocculare was detected, and reactivity with serum from another pig hyperimmunised with killed M hyorhinis showed only weak cross-reactivity, which failed to reach the ELISA positive threshold (ELISA ratio 3) for M hyopneumoniae.     

Mitochondrial Activity, Distribution and Segregation in Bovine Oocytes and in Embryos Produced in Vitro

Contents Bovine oocytes and embryos produced in vitro were studied to determine the mitochondrial pattern of distribution, segregation and activity using DIOC 6 and Jc-1 fluorescence. The highest fluorescence level observed in mature oocytes was taken as 100% activity and six activity levels were estimated as follows: (1) 0%, (2) 1-15%, (3) 16-30%, (4) 31-50%, (5) 51-75% and (6) 76-100%. Three patterns of mitochondrial distribution were found: (1) diffused throughout the cytoplasm in oocytes and embryos, (2) pericytoplasmic in oocytes and embryos, and (3) perinuclear only in embryos. The segregation of mitochondria in blastomeres showed two distinct patterns: (1) symmetrical with an even mitochondrial population, and (2) asymmetrical with different numbers of mitochondria in each blastomere. In immature oocytes, mitochondrial activity was very low and the distribution was diffuse or negligible, while in mature oocytes the activity was high and the distribution was diffuse or pericytoplasmic. Competent embryos up to the 16-cell stage showed intermediate levels of activity (16-50%) but activity decreased thereafter up to the blastocyst stage. Non-competent embryos showed low levels of activity (1-15%) at all stages. These results suggest that mitochondria might play an important role during early development and that a minimum threshold of activity regulates the potential competence for reaching the blastocyst stage.     

Changes in Blood and Semen Quality of Bulls after Artificial Infection with Mycoplasma wenyonii

[目的]验证温氏枝原体对种公牛血液和精液品质的影响。[方法]将10头健康的鲁西黄牛的种公牛随机分成2组,试验组分别于试验当天和第28天静脉注射温氏枝原体,从临床症状、血液涂片镜检、血液生理生化、精液品质和阴囊变化等方面,综合评价温氏枝原体对种公牛的影响。[结果]除个别指标外,试验组临床检查基本正常,未出现明显的血液和精液品质异常。[结论]该研究表明人工感染温氏枝原体对种公牛毒力较低,致病性不强,对血液和精液品质无明显异常。     

Red swamp crayfish,Procambarus clarkii,found in South Africa 22 years after attempted eradication

1. No freshwater crayfish are indigenous to continental Africa, but four species have been introduced to the continent. One of these is the North American red swamp crayfish Procambarus clarkii, which has been introduced into several African countries, mainly for aquaculture, and has had demonstrable impacts where it has escaped captivity. In South Africa, the documentation of this species in farm dams near Dullstroom and the adjacent Crocodile River in 1988 resulted in an eradication attempt in 1994, with unknown results. 2. In order to evaluate the status of P. clarkii in South Africa, dams on the previously invaded farm and the Crocodile River were sampled four times between December 2015 and June 2016 using visual surveys, trapping, dipnetting and electrofishing. This yielded a single reproductively active male P. clarkii from one of the farm ponds, while many other native aquatic species were found in high numbers. 3. It is clear from this study that P. clarkii was not eradicated in South Africa and that individuals have been surviving in the wild (i.e. outside captivity or cultivation) for at least 28 years in the location where it was introduced. Containment and eradication of the species are proposed as management actions, which have major importance in preventing undesirable further spread or translocation of this species into new aquatic environments in South Africa.     

Outbreak of foetal infection with bovine pestivirus in a central Queensland beef herd

OBJECTIVE: To describe a significant outbreak of foetal infection and subsequent losses due to bovine pestivirus on a 5200 ha beef breeding and fattening property in central Queensland. DESCRIPTION OF THE HERD: The affected herd consisted of 656 cows, including 269 recently purchased cows, and 221 heifers that were joined in December/January 1995/96. There were approximately 2500 cattle on the property. INVESTIGATION: Following the purchase of 269 cows in October 1995, which were mingled with the existing cow herd, losses were experienced due to foetal infection with bovine pestivirus. These losses were recorded between 1996 and 1999 as: reduced pregnancy rates, losses between pregnancy testing (midpregnancy) and branding (calves averaged 3 months-of-age), losses due to pneumonia and ill-thrift between branding and approximately 12 months-of-age, and losses due to ill-thrift and the chronic wasting form of mucosal disease thereafter. All surviving calves were tested for bovine pestivirus in 1997 at an average of 10 months. Fifty-three calves were identified as persistently infected with bovine pestivirus. A further 110 calf losses could reasonably be attributed to bovine pestivirus infection. Persistently infected cattle were always unthrifty compared to their virus negative counterparts. Only one persistently infected calf was identified, on the basis of severe ill thrift, in the 1997 birth cohort and none in 1998. CONCLUSIONS: This outbreak of foetal infection with bovine pestivirus resulted in significant production losses. These losses were recorded over the three years subsequent to the outbreak. Significant numbers of persistently infected calves were not evident among calves born in the two years after this outbreak.