Effect of Different Manganese Concentrations during in vitro Maturation of Bovine Oocytes on DNA Integrity of Cumulus Cells and Subsequent Embryo Development

Manganese (Mn) is a trace element present in forages and cereals, and its concentration depends on soil status. Manganese deficiency in cattle, goats and ewes not only impairs oestrous cycle but reduces calf birth weight. The achievement of the first oestrus is delayed, and more attempts are necessary to obtain a successful conception. This study was conducted to investigate the effect of the availability of supplemental Mn during IVM on DNA damage of cumulus cells and total glutathione (GSH) content in oocytes and cumulus cells. The effect of supplementary Mn during IVM on subsequent embryo development was also studied. The results reported here indicate (i) DNA damage in cumulus cells decreased with 0, 2, 5 and 6 ng/ml Mn supplementation during IVM (p < 0.05). (ii) Intracellular GSH‐GSSG content increased (p < 0.01) with different Mn concentrations in oocytes and cumulus cells. Also, cumulus cell number per cumulus oocyte‐complexes (COC) did not differ either before or after IVM. (iii) Addition of Mn to maturation medium resulted in similar cleavage rates (p > 0.05) at 0, 2, 5 and 6 ng/ml Mn. However, subsequent embryo development to blastocyst stage was significantly higher (p < 0.01) in oocytes matured with 5 and 6 ng/ml Mn. (iv) There was also an increase (p < 0.05) in mean cell number per blastocyst obtained from oocytes matured with 5 and 6 ng/ml respect to zero Mn (IVM alone) and 2 ng/ml Mn. This study provides evidence that optimal embryo development to the blastocyst stage was partially dependent on the presence of Mn during IVM. Moreover, the availability of Mn during oocyte maturation ensures ‘normal’ intracellular GSH content in COCs and protects DNA integrity of cumulus cells.     

Abortions in sheep caused by Salmonella Brandenburg: Pathological findings

CASE HISTORY: A 7-year-old cat developed sporadic vomiting, reduced appetite, and weight loss over the previous 3 months.

CLINICAL FINDINGS: Palpation revealed a large mid-abdominal mass and the cat had marked eosinophilia. The cat progressively lost weight over the next 7 weeks when euthanasia was performed.

PATHOLOGICAL FINDINGS: Necropsy revealed a 3?cm diameter firm white intramural mass in the colon and another in the pylorus. Mesenteric and cranial mediastinal lymph nodes were firm, pale, and enlarged. Histopathological examination revealed foci of necrosis surrounded by thick dense collagen trabeculae and predominantly eosinophilic inflammation within the intestine and lymph nodes. Marked eosinophilic infiltration of the liver was also present.

DIAGNOSIS: The lesions were consistent with gastrointestinal eosinophilic sclerosing fibroplasia (FGESF).

CLINICAL RELEVANCE: This is the first report of FGESF in a New Zealand cat and the first time lesions of FGESF have been observed in extra-abdominal tissues. Intestinal neoplasia can be clinically identical to FGESF and histopathology is required for differentiation. Evidence suggests that FGESF has a more favourable prognosis than intestinal neoplasia.     

Serological patterns,antibody half-life and shedding in urine of Leptospira spp. in naturally exposed sheep

Abstract

AIMS: To determine within-farm prevalence, longitudinal pattern of exposure measured by serology, antibody titre longevity and point prevalence of shedding in urine of Leptospira borgpetersenii serovar Hardjo and L. interrogans serovar Pomona in naturally infected sheep on a sample of commercial farms in New Zealand.

METHODS: On eight commercial sheep farms, between September 2011 and January 2014, blood samples were collected from 115–217 ewe lambs on each farm, at intervals of 2–11 months. They were analysed by microscopic agglutination test (MAT) for antibodies to L. borgpetersenii serovar Hardjo and L. interrogans serovar Pomona, using a titre cut-point of 48. Urine from 98 animals was tested by quantitative PCR (qPCR). The half-life of antibodies was estimated in 185 sheep for serovar Hardjo and 21 for Pomona, and the seroprevalence and mean titre of animals lost to follow-up was compared with those remaining in the study.

RESULTS: Within-flock seroprevalence for serovar Hardjo reached a maximum at 17–22 months of age, ranging from 79 to 100%. Seroprevalence for serovar Pomona rose above 10% on three farms and increased to 21–54% by 4–14 months. Seroconversions occurred mainly from late autumn to early summer at 7–15 months of age. Seroprevalences ranging from 3 to 76% for serovar Hardjo and 0.5 to 15% for serovar Pomona were observed up to 3 months of age, likely due to maternally derived immunity. The half-life of antibody in response to infection was estimated to be 6.7 (95% CI=5.8–7.9) months for serovar Hardjo and 6.3 (95% CI=4.8–9.0) months for Pomona. The prevalence of sheep with urine positive for leptospires on qPCR on each farm ranged from 11 to 88%. All but one of the qPCR-positive animals were seropositive for serovar Hardjo. On two farms where Pomona exposure was observed, animals that were lost to follow-up had a higher geometric mean titre for serovar Pomona than those remaining in the study.

CONCLUSIONS: This study demonstrated seasonal exposure from autumn to early summer in young sheep, a wide range of within-flock serological and shedding prevalence, and gives an estimation of the half-life of MAT titres in sheep. More extensive data are needed to fully understand the epidemiology of leptospirosis in sheep flocks across New Zealand and, along with economic analysis, to justify and design cost-effective and efficient control measures to protect human and animal health.     

Intratesticular lidocaine reduces the response to surgical castration in dogs

ObjectiveTo investigate whether intratesticular injection of lidocaine pre-surgery would reduce the intraoperative responses to elective castration in dogs.Study designDouble-blinded, randomized, controlled, prospective clinical study.AnimalsForty-two client-owned dogs weighing 2.2–38.4 kg and aged between 4.5 and 56 months.MethodsGroup L dogs received an intratesticular injection of 2% lidocaine (2 mg kg^?1) and Group S an identical volume of saline prior to surgery. Premedication was with acepromazine and morphine intramuscularly. Anaesthesia was induced with propofol intravenously and maintained with isoflurane vaporized in oxygen. Heart rate (HR), mean arterial pressure (MAP), respiratory rate (f_R), end-tidal isoflurane (Fe′ISO) and carbon dioxide concentrations, oxygen saturation and ECG were monitored during surgery. Fe′ISO was maintained at 1.0 ± 0.1%. Supplemental propofol was given in response to gross movement.ResultsGroup L had significantly lower maximum values for both HR and MAP. Group L displayed significantly smaller increases in HR during exteriorization of the first testis than Group S. There was an overall significant difference in MAP between groups during all surgical events (p = 0.041) and time points (p = 0.002). In univariate analysis, Group L showed significantly less changes in MAP during skin incision, exteriorization of the first testis and clamping of both spermatic cords. Group S reached its highest f_R significantly earlier. Group L (eight dogs) required additional propofol 33 ± 18 minutes after the start of surgery and Group S (seven dogs) at 19 ± 17 minutes; this difference was not statistically significant. Seven dogs in Group L and 12 dogs in Group S required rescue analgesia with morphine (GCMPS-SF score ≥6); this difference was not statistically significant. No adverse effects were reported postoperatively.Conclusions and clinical relevanceBased on this study, the authors recommend the use of intratesticular lidocaine for surgical castration in dogs.     

The sedative effects of low-dose medetomidine and butorphanol alone and in combination intravenously in dogs

ObjectiveTo evaluate the sedative effects of intravenous (IV) medetomidine (1 μg kg^?1) and butorphanol (0.1 mg kg^?1) alone and in combination in dogs.Study designProspective, blinded, randomized clinical trial.AnimalsSixty healthy (American Society of Anesthesiologists I) dogs, aged 6.2 ± 3.2 years and body mass 26 ± 12.5 kg.MethodsDogs were assigned to four groups: Group S (sodium chloride 0.9% IV), Group B (butorphanol IV), Group M (medetomidine IV) and Group MB (medetomidine and butorphanol IV). The same clinician assessed sedation before and 12 minutes after administration using a numerical scoring system in which 19 represented maximum sedation. Heart rate (HR), respiratory rate, pulse quality, capillary refill time and rectal temperature were recorded after each sedation score assessment. Sedation scores, sedation score difference (score after minus score before administration) and patient variables were compared using one-way anova for normally distributed variables and Kruskal–Wallis test for variables with skewed distributions and/or unequal variances. Where significance was found, further evaluation used Bonferroni multiple comparisons for pair-wise testing.ResultsBreed, sex, neuter status, age and body mass did not differ between groups. Sedation scores before substance administration were similar between groups (p = 0.2). Sedation scores after sedation were significantly higher in Group MB (mean 9.5 ± SD 5.5) than in group S (2.5 ± 1.8) (p < 0.001), group M (3.1 ± 2.5) (p < 0.001) and group B (3.7 ± 2.0) (p = 0.003). Sedation score difference was significantly higher in Group MB [7 (0–13)] than in Group S [0 (?1 to 4)] (p < 0.001) and Group M [0 (0–6)] (p < 0.001). HR decreased significantly in Groups M and MB compared with Group S (p < 0.05).Conclusion and clinical relevanceLow-dose medetomidine 1 μg kg^?1 IV combined with butorphanol 0.1 mg kg^?1 IV produced more sedation than medetomidine or butorphanol alone. HR was significantly decreased in both medetomidine groups.     

Improved Prediction of Ovulation Time may Increase Pregnancy Rates to Artificial Insemination in Lactating Dairy Cattle

A prospective observational study was conducted in two Australian dairy herds to assess the potential for improving pregnancy rates (proportions of inseminations that result in pregnancy) to artificial insemination (AI) if the time of ovulation could be predicted with more certainty. Herd 1 calved year‐round and inseminations were performed during two periods each day. Herd 2 calved during autumn–winter and inseminations were performed only after the morning milking each day. In both herds, the AI to ovulation interval of enrolled cows was determined by trans‐rectal ovarian ultrasonography approximately 0, 12, 24 and 36 h after AI, and pregnancy was assessed by palpation per rectum 35–56 days after AI. Also, in Herd 1 vaginal electrical resistance (VER) measurements were taken at approximately 0, 12, 24 and 36 h after AI, and in Herd 2 cows were fitted with neck mounted activity meters that monitored cow activity count in 2‐h periods. There was substantial variation in the intervals from AI to ovulation within and between herds (mean ± SD 21.2 ± 10.7, n = 102; 14.7 ± 10.4, n = 100 in herds 1 and 2, respectively). Pregnancy rates were higher for inseminations close to, but preceding, ovulation. Using combined herd data (n = 202), the highest pregnancy rate (50.8%) was observed for inseminations between 0 and 16 h before ovulation, a period in which only a modest proportion of inseminations (31.2%) occurred. In contrast, pregnancy rate was significantly lower (28.7%; risk ratio 0.6; 95% CI 0.4–1.0; p = 0.039) for inseminations between 16 and 32 h before ovulation, a period where the highest proportion of inseminations (53.2%) occurred. Thus pregnancy rates could potentially be improved if a greater proportion of inseminations were conducted shortly before ovulation. In Herd 1, mean VER during the peri‐ovulatory period varied with time from ovulation. Lowest values (mean ± SEM, VER = 64.8 ± 1.2, n = 55) occurred approximately 18 h before ovulation and were significantly lower than measurements approximately 6 h before ovulation (67.4 ± 1.0; n = 73; p = 0.003). Further work is required to determine if VER can be used to identify ovulation time and hence the optimal time to inseminate in individual animals. In Herd 2 a modest proportion of inseminations (26.9%) occurred between 24 and 40 h after the onset of increased cow activity where the highest pregnancy rate (67.9%) was observed, whereas a significantly lower pregnancy rate (42.4%; risk ratio 0.6; 95% CI 0.4–0.9; p = 0.036) was observed for inseminations between 8 and 24 h after the onset of increased cow activity where the highest proportion of inseminations (56.7%) occurred. Thus cow activity monitoring may be useful to identify the optimal time to inseminate cows. Results from this study indicate that improved methods of ovulation prediction may allow better insemination timing relative to ovulation and consequently increased pregnancy rates.     

Inherited epidermal dysplasia in Holstein-Friesian calves

Inherited epidermal dysplasia (IED), formerly called baldy calf syndrome, is a lethal disease of calves of Holstein-Friesian ancestry. The disease causes progressive illthrift and skin, horn and hoof lesions, which can be confused with inherited zinc deficiency. The clinicopathological features and ancestry of 10 affected calves in Gippsland, Victoria are described.