The Importance of Having Zinc During In Vitro Maturation of Cattle Cumulus–Oocyte Complex: Role of Cumulus Cells

The aim of this study was to investigate the influence of zinc (Zn) on the health of cumulus–oocyte complex (COC) during in vitro maturation (IVM). Experiments were designed to evaluate the effect of Zn added to IVM medium on: DNA integrity, apoptosis, cumulus expansion and superoxide dismutase (SOD) activity of cumulus cells (CC). Also, role of CC on Zn transport during IVM was evaluated on oocyte developmental capacity. DNA damage and early apoptosis were higher in CC matured with 0 μg/ml Zn compared with 0.7, 1.1 and 1.5 μg/ml Zn (p < 0.05). Cumulus expansion did not show differences in COC matured with or without Zn supplementation (p > 0.05). Superoxide dismutase activity was higher in COC matured with 1.5 μg/ml Zn than with 0 μg/ml Zn (p < 0.05). Cleavage and blastocyst rates were recorded after IVM in three maturation systems: intact COCs, denuded oocytes with cumulus cells monolayer (DO + CC) and denuded oocytes (DO). Cleavage rates were similar when COC, DO + CC or DO were matured with 1.5 μg/ml Zn compared with control group (p > 0.05). Blastocyst rates were significantly higher in COC than in DO + CC and DO with the addition of 1.5 μg/ml Zn during IVM (p < 0.01). Blastocyst quality was enhanced in COC and DO + CC compared with DO when Zn was added to IVM medium (p < 0.001). The results of this study indicate that Zn supplementation to IVM medium (i) decreased DNA damage and apoptosis in CC; (ii) increased SOD activity in CC; (iii) did not modify cumulus expansion and cleavage rates after in vitro fertilization; (iv) improved subsequent embryo development up to blastocyst stage; and (v) enhanced blastocyst quality when CC were present either in intact COC or in coculture during IVM.     

Risk of Infectious Gastroenteritis in Young Children Living in Québec Rural Areas with Intensive Animal Farming: Results of a Case–Control Study (2004–2007)

This study was designed to evaluate the epidemiology of severe gastroenteritis in children living in Québec rural areas with intensive livestock activities. From September 2005 through June 2007, 165 cases of gastroenteritis in children aged from 6 months to 5 years, hospitalized or notified to the public health department were enrolled, and 326 eligible controls participated. The parents of cases and controls were asked questions about different gastroenteritis risk factors. The quality of the drinking water used by the participants was investigated for microbial indicators as well as for four zoonotic bacterial pathogens (Campylobacter spp, Escherichia coli, Salmonella spp and Yersinia spp) and two enteric parasites (Cryptosporidium spp and Giardia spp). From 134 stool specimen analysed, viruses were detected in 82 cases (61%), while 28 (21%) were found with at least one of the bacteria investigated, and five cases were infected by parasites. Campylobacteriosis was the main bacterial infection (n = 15), followed by Salmonella sp (n = 7) and E. coli O157:H7 (n = 5) among cases with bacterial gastroenteritis. No significant difference was found between cases and controls regarding the quality of water consumed; the frequency of faecal contamination of private wells was also similar between cases and controls. Considering the total cases (including those with a virus), no link was found between severe gastroenteritis and either being in contact with animals or living in a municipality with the highest animal density (4th quartile). However, when considering only cases with a bacterial or parasite infection (n = 32), there was a weak association with pig density that was not statistically significant after adjusting for potential confounders. Contact with domestic, zoo or farm animals were the only environmental factor associated with the disease.     

Effects of tramadol,morphine or their combination in dogs undergoing ovariohysterectomy on peri-operative electroencephalographic responses and post-operative pain

Abstract

AIM: To compare the peri-operative electroencephalogram (EEG) responses and post-operative analgesic efficacy of pre-operative morphine or tramadol with a combination of low-dose pre-operative morphine and post-operative tramadol, in dogs undergoing ovariohysterectomy.

METHODS: Dogs undergoing routine ovariohysterectomy were treated with either pre-operative morphine (0.5 mg/kg S/C, n=8), or tramadol (3 mg/kg S/C, n=8), or pre-operative low-dose morphine (0.1 mg/kg S/C) and post-operative tramadol (3 mg/kg I/V, n=8). All dogs received routine pre-anaesthetic medication, and anaesthesia was induced with I/V thiopentone to effect and maintained with halothane in oxygen. Respiratory rate, heart rate, end-tidal halothane tension (EtHal) and end-tidal CO₂ tension (EtCO₂) were monitored throughout surgery. The EEG was recorded continuously in a three electrode montage. Median frequency (F50), total power (Ptot) and 95% spectral edge frequency (F95) of the EEG power spectra were compared during different 100-second periods of surgery: prior to and during skin incision, ligation of each ovarian pedicle, ligation of uterine body and skin closure. Post-operatively, pain was assessed using the short form of the Glasgow composite measure pain scale (CMPS-SF).

RESULTS: There was no difference in F50 or Ptot of the EEG between baseline and noxious surgical events within each treatment group, or between the three groups (p>0.05). The mean F95 was higher during the first three periods of surgery for dogs administered tramadol and low-dose morphine than those that received 0.5 mg/kg morphine (p=0.001). Dogs that received low-dose morphine and tramadol had lower CMPS-SF pain scores after ovariohysterectomy than those that received either tramadol or morphine alone (p=0.001). There was no difference in pain scores between dogs in the latter two groups.

CONCLUSION AND CLINICAL RELEVANCE: Tramadol and morphine administered pre-operatively provided an equal degree of post-operative analgesia in dogs after ovariohysterectomy. A combination of pre-operative low-dose morphine and post-operative tramadol produced better post-operative analgesia than either drug administered alone pre-operatively. Administration of analgesics pre- and post-operatively could result in improved post-operative well-being of ovariohysterectomised dogs.     

Risk characterisation and management of sewage sludge on agricultural land-implications for the environment and the food-chain

The disposal of sewage wastes may cause severe environmental problems as was graphically demonstrated with pollution on Sydney's ocean beaches in recent years. Sewage sludges contain valuable plant nutrients and organic matter which can improve the fertility and structure of the soil. However, human parasites, pathogenic micro-organisms and chemicals capable of causing soil contamination, phytotoxicity and residues in animal products may also be present. Although sewage sludge is frequently spread on agricultural land overseas, it is not common in Australia and most states do not have specific regulations to minimise risk and promote good practice. A sludge-to-land program began in the Sydney region in 1990. It follows guidelines written by NSW Agriculture to encourage beneficial agricultural use of sludge by adoption of environmentally sustainable practices. This article describes the major risks to the food-chain and the environment, which may be associated with applying sewage sludge to agricultural land. It summarises how the risks are managed, and where further research data are required.     

Primary anconeal fracture in a boxer

A 14-month-old speyed female Boxer was presented with acute non-weight-bearing lameness in the right forelimb. Radiography revealed separation of the anconeal process, which was thence surgically removed. Histological examination of the anconeal process confirmed a primary fracture. Isolated unilateral anconeal process fracture in the dog is rare and this report includes histopathological findings of a primary anconeal fracture.     

In vitro 3-dimensional kinematic evaluation of 2 corrective operations for cranial cruciate ligament-deficient stifle

The objective of this study was to determine the immediate postoperative effect of 2 corrective operations for cranial cruciate ligament (CCL)-deficient stifle by evaluating 3-dimensional (3-D) stifle kinematics. Ten hindlimbs from large-breed canine cadavers were used. Range of motion was induced by applying 100 N of traction on the quadriceps tendon and recorded with electromagnetic movement sensors for each situation: intact stifle (control), CCL-sectioned stifle, and surgical correction of the sectioned ligament with the modified retinacular imbrication technique (MRIT) and then with a tibial plateau leveling osteotomy--Montavon (TPLO-M). The results for the experimental situations were compared with the results for the control situation by 1-way repeated-measures analysis of variance and with each other by post-hoc analysis with the least-significant-difference method. Range of motion was significantly decreased by MRIT as compared with the other situations. Normal cranial tibial translation was restored after MRIT, whereas TPLO-M resulted in significant caudal translation. A significant increase in external rotation was observed after both MRIT and TPLO-M. A significant increase in tibial adduction throughout the range of motion was observed with TPLO-M, whereas a significant increase in tibial abduction was observed after MRIT. This study allowed us to better understand objectively the effects on 3-D canine stifle kinematics of MRIT and TPLO-M. We suggest that this type of in vitro study would be useful to evaluate established and upcoming surgical techniques and potentially improve corrective surgery.     

Breeding Soundness Evaluation and Semen Analysis for Predicting Bull Fertility

Bull fertility is influenced by numerous factors. Although 20–40% of bulls in an unselected population may have reduced fertility, few are completely sterile. Breeding soundness refers to a bull's ability to get cows pregnant. A standard breeding soundness evaluation identifies bulls with substantial deficits in fertility, but does not consistently identify sub-fertile bulls. In this regard, the use of frozen-thawed semen (from bulls in commercial AI centres) that meets minimum quality standards can result in pregnancy rates that differ by 20–25 percentage points. Although no single diagnostic test can accurately predict variations in fertility among bulls that are producing apparently normal semen, recent studies suggested that a combination of laboratory tests were predictive of fertility. This review is focused on recent developments in prediction of bull fertility, based on assessments at the molecular, cellular and whole-animal levels.     

Co‐culture of Buffalo Preantral Follicles with Different Somatic Cells

The effect of co‐culture of buffalo preantral follicles (PFs) with different somatic cells, i.e, cumulus, granulosa, ovarian mesenchymal and oviductal epithelial cells was studied. Large PFs (250–450 μm) were isolated by microdissecting the trypsin (1%) digested ovarian cortical slices. Cumulus cells were isolated by repeated pipetting of oocytes, granulosa cells were isolated by aspirating from punctured PFs and ovarian mesenchymal cells were isolated from ovarian cortex by scraping the cortical slices and passing through 20 μm filter. Preantral follicles were cultured in standard culture medium without somatic cells or co‐cultured with cumulus cells, granulosa cells, ovarian mesenchymal cells and oviductal epithelial cells for 80 days. The growth rate (μm/day) of the PFs was monitored by measuring follicular diameter on day 0, 30, 60 and 80 days of culture. The viability of PFs was evaluated by trypan blue staining. The results indicated that PFs co‐cultured with cumulus, granulosa and ovarian mesenchymal cells had a better development and survivality compared with control and those co‐culture with oviductal epithelial cells. Maximum growth and survivality of PFs were achieved when cultured with cumulus cells. It is concluded that inclusion of somatic cells in PF culture media had beneficial effect on the growth of PFs and cumulus cells supported maximum growth and survivality of PFs in vitro of all somatic cells tested.     

Bovine thyroglobulin gene polymorphisms and their association with sexual precocity in Guzerat bulls

Puberty is a stage of sexual development determined by the interaction of environmental factors and genetic mechanisms. Among them, thyroid function plays a key role in sexual development and spermatogenic function and is under the control of several genes, including the well‐described thyroglobulin gene (TG). Previous reports have shown genetic association between thyroid function and selected single nucleotide polymorphisms (SNPs) in taurine cattle. Therefore, the identification of genetic mechanisms involved in the regulation of this trait can assist with the selection for early pubertal bulls, thus improving genetic progress in livestock breeding. The aim of this study was to validate the association between TG SNPs and age at puberty in zebuine bulls. Three SNPs (rs110406764, rs109662686, rs109057985) were genotyped in 159 Guzerat animals using SEQUENOM technology. Results showed a significant association (p < .05) between the studied SNPs and puberty age, in agreement with our previous reports in a taurine breed. Interestingly, allele frequencies were different from those already reported, being GAT the most favourable allele for age at puberty in Guzerat (94.4 days lower). Overall, our findings corroborate previous reports and reinforce the importance of genetic influence in the regulation of sexual development and puberty through a thyroid pathway in zebuine cattle.