Manipulated Mouse Embryos as Bioassay System for Water Quality Control

Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K⁺ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D‐O) water and Milli‐Q system (M‐Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA‐supplemented protein‐free culture media, whereas in EDTA‐ and protein‐free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M‐Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein‐free culture media were very low (0–7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M‐Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system.     

ACE inhibition in goats under fixed‐time artificial insemination protocol increases the pregnancy rate and twin births

To assess the effect of the angiotensin‐converting enzyme (ACE) inhibition on the efficiency of the fixed‐time artificial insemination (TAI), 69 goats were divided randomly into two groups: enalapril (n = 35) and control (n = 34). In the experiment, all animals underwent the protocol of fixed‐time artificial insemination for 12 days. Enalapril group received enalapril maleate dissolved in saline (Enalapril, Lab Teuto Ltda) subcutaneously at the following doses: 0.2 mg/kg/day in D0‐D2; 0.3 mg/kg/day in D3‐D6 and 0.4 mg/kg/day in D7‐D11. The control group received the corresponding volume of 0.9% saline solution. We performed a single insemination 36 hr after sponge removal using frozen semen from two adult male goats with recognized fertility. The ultrasound pregnancy diagnosis was 30 days after the artificial insemination (AI). There was significant increase in pregnancy rates and twinning as well as a decrease in foetal loss in animals receiving enalapril (p < .01). The use of ACE inhibitors during the TAI protocol was shown to be a promising alternative to increase the efficiency of such reproductive biotechnology.     

Gonadotropin‐induced Puberty Does Not Impair Reproductive Performance of Gilts over Three Parities

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty‐nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post‐weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning‐to‐oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.     

Preserving new anthelmintics: a simple method for estimating faecal egg count reduction test (FECRT) confidence limits when efficacy and/or nematode aggregation is high

As it has been 30 years since a new anthelmintic class was released, it is appropriate to review management practices aimed at slowing the development of anthelmintic resistance to all drug classes. Recommendations to delay anthelmintic resistance, provide refugia and the use of a simulation model were reviewed to find optimum treatment strategies that maintain nematode control. Simulated Australian conditions indicated that a common successful low-risk treatment program was a rapid rotation between a "triple-combination" product (benzimidazole+levamisole+abamectin) and a new high-efficacy drug (monepantel). Where Haemonchus contortus was a threat, moxidectin was required at critical times because of its persistent activity against this parasite. Leaving up to 4% of adult sheep untreated provided sufficient "refugia" for non-selected worms to reduce the risk of selecting for anthelmintic resistance without compromising nematode control. For a new anthelmintic, efficacy estimated by faecal egg count reduction (FECR) is likely to be at or close to 100%, however using current methods the 95% confidence limits (CL) for 100% are incorrectly determined as 100%. The fewer eggs counted pre-treatment, the more likely an estimate of 100% will occur, particularly if the true efficacy is >90%. A novel way to determine the lower-CL (LCL) for 100% efficacy is to reframe FECR as a binomial proportion, i.e. define: n and x as the total number of eggs counted (rather than eggs per gram of faeces) for all pre-treatment and post-treatment animals, respectively; p the proportion of resistant eggs is p = x/n and percent efficacy is 100 ×(1-p) (assuming equal treatment group sizes and detection levels, pre- and post-treatment). The LCL is approximated from the cumulative inverse beta distribution by: 95%LCL=100 ×(1-(BETAINV(0.975, x+1, n-x+1))). This method is simpler than the current method, independent of the number of animals tested, and demonstrates that for 100% efficacy at least 37 eggs (not eggs per gram) need to be counted pre-treatment before the LCL can exceed 90%. When nematode aggregation is high, this method can be usefully applied to efficacy estimates lower than 100%, and in this case the 95% upper-CL (UCL) can be estimated by: 95% UCL = 100 ×(1((BETAINV(0.025, x+1, n-x+1))), with the LCL approximated as described above. A simulation study to estimate the precision and accuracy of this method found that the more conservative 99%CL was optimum; in this case 0.975 and 0.025 are replaced by 0.995 and 0.005 to estimate the LCL and UCL, respectively.     

Postpartum Ovarian Activity in South Asian Zebu Cattle

Timely onset of postpartum ovarian activity is vital for optimal reproductive performance of dairy cows. Much depends upon genetic constitution of an animal although several factors interplay to govern the onset of postpartum ovarian activity. South Asian zebu cattle have much longer service period when compared with other exotic or crossbred cattle reared in the same Asian environment, which suggests differences in their genetic makeup. However, the cows with same genetic configuration expressed better reproductive potential when reared under different environment, such as in Brazil and Mexico, which suggests the role of extrinsic factors such as management, nutrition, environment and disease conditions. Better management of animals (provision of proper shade, water and housing, efficient oestrous detection and timely insemination), good quality nutrition supplemented with appropriate minerals and vitamins, prevention of diseases (vaccination, deworming, suitable therapeutic interventions) and application of biotechnology have helped in improving postpartum ovarian activity and, therefore, reproductive performance of zebu cattle in Asia. No comprehensive study appears to have been carried out on the various aspects of reproduction in zebu cattle reared under South Asian socio-agro-climatic conditions. This paper is a modest effort to collect what ever information available and to critically review the postpartum ovarian activity in zebu cattle with special reference to the effect of the various managemental practices and pharmacological interventions.     

Outcomes of Heimlich valve drainage in dogs

Objective and design   Retrospective study of the outcomes of Heimlich valve drainage in dogs.
Procedure   Medical records of the past 3 years were retrospectively reviewed. Heimlich valve drainage was used in 34 dogs (median body weight 30 ± 5 kg): lobectomy (n = 15), pneumonectomy (n = 9), intrathoracic oesophageal surgery (n = 2), diaphragmatic hernia repair (n = 1), traumatic open pneumothorax (n = 2), bilobectomy (n = 2), ligation of the thoracic duct (n = 1), and chylothorax and pneumothorax (n = 1 each). Evacuation of air and/or fluid from the pleural cavity was performed with the Heimlich valve following thoracostomy tube insertion. During drainage, the dogs were closely monitored for possible respiratory failure. Termination of Heimlich valve drainage was controlled with underwater seal drainage and assessed with thoracic radiography.
Results   Negative intrathoracic pressure was provided in 29 dogs without any complications. Post pneumonectomy respiratory syncope and post lobectomy massive hemothorax, which did not originate from the Heimlich valve, were the only postoperative complications. Dysfunction of the valve diaphragm, open pneumothorax and intrathoracic localisation of an acute gastric dilatation–volvulus syndrome caused by a left-sided diaphragmatic hernia following pneumonectomy were the Heimlich valve drainage complications.
Conclusions   The Heimlich valve can be used as a continuous drainage device in dogs, but the complications reported here should be considered by veterinary practitioners.