A critical evaluation of serum methylmalonic acid and vitamin B12 for the assessment of cobalt deficiency of growing lambs in New Zealand

AIM: To derive reference ranges for serum methylmalonic acid (MMA) for the diagnosis of cobalt/vitamin B₁₂-responsiveness in lambs and critique existing serum vitamin B₁₂ reference ranges.

METHODS: Individual animal data from earlier supplementation trials, involving 225 ewes, 106 suckling lambs, 301 lambs during the suckling and post-weaning periods and 414 weaned lambs, for which weight gain to supplementation was observed, were used to derive relationships between serum vitamin B₁₂ and MMA, and liveweight gain.

RESULTS: Serum MMA concentrations were rarely elevated above the norm of <2 µmol/L when serum vitamin B₁₂ concentrations were >375 pmol/L, and not elevated into the range where a liveweight response to supplementation occurred (>10 µmol/L) unless serum vitamin B₁₂ concentrations were below 200 pmol/L. Suckling lambs were able to maintain high growth rates despite elevated serum MMA concentrations (>20 µmol/L).

CONCLUSIONS: The current reference ranges used in New Zealand for serum vitamin B₁₂ are set conservatively high. Serum MMA concentrations appear to allow better differentiation of a responsive condition than vitamin B₁₂ concentrations. Serum MMA concentrations <13 µmol/L indicate responsiveness to supplementation whilst concentrations <7 µmol/L indicate unresponsiveness. In the range 7–13 µmol/L, variation in response was observed and predictability of response is less certain, but supplementation is advisable.

CLINICAL RELEVANCE: The current reference ranges for vitamin B₁₂ responsiveness are conservatively high and lead to over-diagnosis of vitamin B₁₂ deficiency in ill-thriftiness of sheep.     

In vitro Fertilizing Capacity of Frozen-thawed Bull Spermatozoa Selected by Single-layer (Glycidoxypropyltrimethoxysilane) Silane-coated Silica Colloidal Centrifugation

Barriers to the use of density gradient centrifugation for preparing animal spermatozoa for artificial insemination (AI) include the scarcity of animal-specific formulations and the daunting prospect of processing large volumes of ejaculate in small aliquots (1.5 ml extended semen). Recently, new colloid formulations have been tested in vitro in a modified procedure, centrifugation on a single layer of colloid. The present study investigated the fertilizing ability during in vitro fertilization (IVF) of frozen-thawed bovine spermatozoa following centrifugation through a single layer of glycerolpropylsilane (GS)-coated silica colloid with a species-specific formulation (patent applied for; treatment, T). Controls (C) included centrifugation through gradients of either the same colloid (C1) or Percoll™ (C2). Sperm recovery surpassed 50% for both C1–C2 and T (n.s.). Mean values of various parameters of computerized analysis of sperm motility did not differ between T and C1 (n.s.), and only the proportions of path straightness and linearity were lower in T vs C2 (p < 0.05). In T, the mean (±SD) percentages of fertilization rate, blastocyst development rate and the total number of blastomeres were 58.1 ± 23.3%, 24.5 ± 14.3% and 94.6 ± 23.4%, respectively. The proportions did not differ significantly from controls (C1/C2). Therefore, centrifugation through a single layer of colloid offers an alternative method to density gradient centrifugation for selection of viable, potentially fertile frozen-thawed bull spermatozoa. This single-layer technique is gentle, versatile and convenient because it facilitates scaling-up the process of sperm preparation to allow larger numbers of spermatozoa (for instance, whole ejaculates) to be processed for AI.     

The effect of ivermectin treatment of late pregnant dairy cows in south-west Victoria on subsequent milk production and reproductive performance

SUMMARY A total of 498 dairy cows in 5 predominantly pasture-fed herds were allocated to pairs. One cow in each pair was treated with a single dose of ivermectin during the dry period. Treated and untreated cows were managed as a single group throughout the trial. Most cows calved between 45 and 115 days after treatment. When data from all herds were pooled, treated cows produced an extra 74 L of milk over the first 100 days of the subsequent lactation (95% confidence interval 20 to 128). Means were greater among treated groups relative to untreated groups in all 5 herds. However, when analysed individually, differences were statistically significant (P < 0.05) in 1 herd only. Over the complete lactation, mean milk volume for treated cows was 86 L greater than for untreated cows (95% confidence interval of difference -57 to 229; P = 0.24). Untreated cows produced 2473 L and 5883 L for the first 100 days of lactation and for the complete lactation, respectively. Milk production responses to treatment did not vary significantly with parity, body condition score, previous production index, calving date category or with plasma pepsinogen concentration or faecal egg count at the time of treatment. Faecal egg counts and plasma pepsinogen concentrations were low at the start of the study. The interval from calving to conception was 4.8 days less in treated cows (95% confidence interval 1.2 to 8.2) relative to untreated cows when data from all 5 herds were pooled. Differences within individual herds were not statistically significant.     

Evidence that moxidectin is a greater risk factor than ivermectin in the development of resistance to macrocyclic lactones by Ostertagia spp in sheep in south eastern Australia

AIM: To determine associations between resistance of Ostertagia (= Teladorsagia) spp to macrocyclic lactone (ML) anthelmintics and history of use of anthelmintics, by type, on commercial sheep farms in temperate regions of southern South Australia and Victoria, Australia.

METHODS: Faecal egg count reduction tests (FECRTs) were conducted during a 2.5-year period (from August 2001 to January 2004) and records of the type of anthelmintic used in the 5 years preceding the FECRTs were collected from commercial sheep farms (n=103) in southern South Australia and Victoria, and data analysed retrospectively. ML resistance was defined as <95% reduction of Ostertagia spp 10–14 days after treatment with ivermectin (IVM), orally, at half the manufacturer's recommended dose rate. Use of anthelmintics in the preceding 5 and 10 years on each property was classified according to the nett number of years each of the following classes of drug had been used: IVM oral liquid (IVO), IVM controlled-release capsules (CRCs), abamectin (ABA), moxidectin (MOX) or a non-ML an- thelmintic. The prevalence of ML resistance, by property, was analysed for associations with prior use of anthelmintics.

RESULTS: Resistance by Ostertagia spp to ML anthelmintics was evident on 51/103 (49.5%) properties. The prevalence of resistance was lowest (23%) on properties on which MOX had not been used, and was significantly higher (64–77%) on properties on which MOX had been used for ≥2 of the preceding 5 years (p<0.001). In contrast, the prevalence of resistance was highest (70–74%) on the properties on which IVM, or IVM and/ or ABA, had not been used in the previous 5 years (on which the use of MOX was predominant), and was markedly lower (20– 42%) on properties that had used IVM or IVM and/or ABA for at least one of the preceding 5 years. Prevalence of resistance was higher for properties on which the only ML anthelmintic used was MOX (19/29=66%) than for those on which the only ML used was IVO (2/19=11%; p<0.001). Properties on which the only ML used was MOX were 2.72 times more likely to have resistance than properties on which the only ML used was IVO (95% confidence interval (CI) = 1.01–5.08).

CONCLUSION: Use of MOX for ≥2 of the preceding 5 years was associated with a higher prevalence of resistance to ML by Ostertagia spp on sheep farms in south eastern Australia than the use of IVO.     

Disease signs reported in south-eastern Australian dairy cattle while grazing Brassica species

Objective
To estimate the relative frequency of various disease signs in dairy cattle fed brassicas in south-eastern Australia, to estimate the incidence of disease signs within affected groups and to identify risk factors for commonly reported signs in brassica-fed dairy cows.
Design
A case series study and a case-control study.
Procedures
Case data was collected using an incident reporting system. Herd managers and veterinarians recorded details about disease incidents in brassica-fed dairy cattle in summer and autumn 1995 using a standard questionnaire. Potential risk factors for photosensitisation were assessed using a case-control study.
Results
Disease signs were reported in 66 groups of brassica-fed dairy cattle. Photosensitisation and bloat were the more frequently reported signs. While high incidences were reported in some groups, the incidence of disease signs was low within most affected groups. Photosensitisation occurred more frequently among groups of cows fed brassica crops treated with nitrogenous fertilisers or which were low yielding. Risks of other disease signs were greatest while the first one quarter of the crop was grazed.
Conclusions
In the study population, most disease outbreaks occurring in brassica-fed cattle were of low incidence. However, some high incidence outbreaks occurred. Results from this study suggest that important risk factors exist for disease signs in brassica-fed dairy cattle. Further studies are required if these risk factors are to be fully identified. This would allow the development of preventative strategies for high incidence disease outbreaks while feeding brassicas to dairy cattle.     

Motility Characteristics and Fertilizing Capacity of Boar Spermatozoa Stained with Hoechst 33342

Flow cytometry sorting of spermatozoa using fluorescence dye Hoechst 33342 is the only effective sex selection methodology validated in numerous laboratories. This study was carried out to determine the effect of Hoechst 33342 on the motility and fertility of stained boar spermatozoa. Experiment 1 evaluated motility parameters (percentage of motile spermatozoa, velocity, angularity and oscillation) of boar spermatozoa stained with Hoechst 33342 by a computer‐aided sperm analysis (CASA) instrument. Spermatozoa (30 million/ml) were divided into five treatment groups and stained during 1 h at 35°C with 9, 18, 27, 60 and 90 μM of H33342. There were no differences in sperm motility patterns nor percentages of motile spermatozoa incubated in the presence of 9, 18 or 27 μM. Percentage of motile spermatozoa and motility parameters decreased significantly (p < 0.05) at 60 μM of Hoechst 33342. Spermatozoa were immotile at concentration of 90 μM. In experiment 2, pregnancy rates, farrowing rates and litter size from sows (n = 275) artificially inseminated (AI) with either Hoechst 33342 stained (27 μM) or unstained (control) spermatozoa were determined. Sows inseminated with stained spermatozoa had no significant lower pregnancy rate (88.33%) as compared with controls (90.32%). Staining neither affected farrowing rates (85.0 vs 87.7%) nor total number of piglets born (10.56 ± 0.32 vs 10.47 ± 0.24, stained and controls, respectively). No phenotypical abnormalities were registered among the newborn piglets. The data suggest that incubating spermatozoa with Hoechst 33342 at levels required for X‐ and Y‐bearing chromosome sperm sorting, does not impair sperm viability or their fertility after AI.     

A screening test for subclinical liver disease in horses affected by pyrrolizidine alkaloid toxicosis

Objective: To evaluate various biochemical tests as indicators of subclinical liver disease in horses exposed to pyrrolizidine alkaloid toxicosis.
Design: A clinical pathology field study.
Animals: Twenty-two clinically normal horses from four properties in the Kimberley region of Western Australia.
Procedure: Serum samples from each horse were assayed for gamma glutamyltransferase, alkaline phosphatase and aspartate aminotransferase activities, and for serum bile acid concentration, albumin and total protein. Serum protein electrophoresis was performed and their amino acid profiles determined. Bromosulphophihalein halfclearance times were measured. Horses were then subjected to a single liver biopsy. Results were analysed by, variance of group means, the Fisher-Irwin exact test, and by sensitivity and specificity calculation.
Results: Horses were classified into 2 groups, of 10 unaffected and 12 subclinically affected, on the basis of liver histology. Significant differences between the unaffected and subclinical groups were observed for gamma glutamyltransferase and alkaline phosphatase activities (P < 0.01). Gamma glutamyltransferase had sufficient sensitivity (75%) and specificity (90%) to function as a primary screening test for subclinical liver disease in horses exposed to pyrrolizidine alkaloids. Alkaline phosphatase was useful, but with lower sensitivity (58%).
Conclusion: Serum gamma glutamyltransferase activity is a useful screening test for detecting subclinical liver disease in horses exposed to pyrrolizidine alkaloids under field conditions in northern Australia.     

Propofol and fentanyl infusions in dogs of various breeds undergoing surgery

ObjectiveTo report the cardiovascular variables, anaesthetic effects and recovery quality of an anaesthesia technique using variable rate infusion propofol combined with constant rate infusion fentanyl in dogs undergoing elective surgery.Study designProspective clinical trial.AnimalsA total of 27 dogs, aged 2.7 ± 2.65 years and weighing 24 ± 11 kg.MethodsFollowing intramuscular acepromazine (0.03 or 0.05 mg kg^?1) and subcutaneous carprofen (4 mg kg^?1) pre-medication, anaesthesia was induced with propofol (4.0 ± 0.5 mg kg^?1) intravenously (IV). All dogs were ventilated with 100% oxygen to maintain normocapnia. Propofol was infused at 0.4 mg kg^?1 minute^?1 for 20 minutes and then at 0.3 mg kg^?1minute^?1. If mean arterial blood pressure (MAP) decreased below 70 mmHg, propofol infusion was reduced by 0.1 mg kg^?1 minute^?1. Five minutes after induction of anaesthesia, fentanyl was administered (2 μg kg^?1) IV followed by the infusion at 0.5 μg kg^?1 minute^?1 and atropine (40 μg kg^?1) IV. Heart rate, MAP, respiratory rate, tidal volume, end-tidal carbon dioxide, presence of reflexes, movements and recovery times and quality were recorded.ResultsMean anaesthetic duration was 131 ± 38.5 minutes. Mean heart rate peaked 10 minutes after atropine injection and gradually declined, reaching pre-anaesthetic values at 55 minutes. MAP easily was maintained above 70 mmHg. Mean times to return of spontaneous ventilation, extubation, head lift and sternal recumbency were 21 ± 10.1, 33 ± 14.6, 43 ± 19.7 and 65 ± 23.4 minutes, respectively. Recovery was smooth and quiet. The time to sternal recumbency was significantly correlated with the duration of anaesthesia and total dose of propofol; time to extubation was correlated to total dose of propofol.Conclusion and clinical relevancePropofol and fentanyl infusions provided stable cardiovascular function and satisfactory conditions for surgery. Some modifications of infusion rates are required to improve the long-recovery times.     

A non-invasive technique for standing surgical repair of urinary bladder rupture in a post-partum mare: a case report

An 11-year-old mare presented 36 hours after foaling with a ruptured bladder. Uroperitoneum was diagnosed on ultrasound and from the creatinine concentration of the peritoneal fluid. Bladder endoscopy demonstrated tissue necrosis and a rent in the dorsocranial aspect of the bladder. Following stabilisation, including abdominal drainage and lavage, the mare was taken to standing surgery. Under continuous sedation and epidural anaesthesia, and after surgical preparation, a Balfour retractor was placed in the vagina. Using sterile lubricant and moderate force, it was possible to insert a hand into the bladder. The tear was easily palpable on the dorsal portion of the bladder. Two fingers were inserted through the tear and used to provide traction to evert the bladder completely into the vagina where it could grasped with the surgeons other hand to prevent further trauma. A second surgeon could then visualise the entire tear and repaired this using a single layer of size zero PDS suture in a single continuous pattern. As soon as the bladder was repaired, it was replaced via the urethra. The mare did well after surgery and was discharged after 48 hours, apparently normal.This report is the first describing repair of the bladder without an abdominal incision or incision into the urethral sphincter. This greatly reduces the chance of possible complications such as urine pooling after surgery with the previously described standing technique or bladder trauma due to traction with abdominal surgery.