Genotyping Brahman cattle for generalised glycogenosis

OBJECTIVE: To develop procedures for genotyping Brahman cattle for loss-of-function alleles within the acidic alpha-glucosidase gene and to assess the risk of generalised glycogenosis in Australian Brahman cattle. DESIGN: PCR assays for three loss-of-function alleles were designed to exploit internal restriction sites within acidic alpha-glucosidase amplicons that are independent of allelic variants at the mutant sites. RESULTS: Genotyping 8529 clinically normal Brahmans between August 1996 and August 2001 revealed 16.4% were heterozygous for the more common of the two mutations (1057deltaTA, often referred to as the 'E7' mutation) that cause generalised glycogenosis in this breed. The less common 1783T mutation (often referred to as the 'E13' mutation) was restricted to descendants of one imported bull, and was not detected in 600 randomly selected Brahmans. Prior to definition of these two disease-causing mutations, 640 (18%), and 14 (0.4%), of 3559 clinically normal Brahmans analysed between January 1994 and December 1996, were heterozygous, and homozygous, respectively, for a silent polymorphism (2223G-->A) that is associated with generalised glycogenosis. In addition to the 1057deltaTA and 1783T mutations, approximately 15% of Brahmans were found to be heterozygous for a single base substitution in exon 9 (1351T, commonly referred to as the 'E9' mutation) that significantly reduces acidic alpha-glucosidase activity, but has not been associated with clinical disease. These three loss-of-function alleles were found in Brahmans imported, or selected for import, from the USA. CONCLUSION: The PCR procedures reported here represent a significant improvement in reliability and accuracy over previous published methods. Utilisation of these PCR/restriction enzyme based assays will facilitate precise selection against the 1057deltaTA and 1783T alleles, and consequently reduce the incidence of generalised glycogenosis in registered and commercial Brahman herds.     

Clinical, diagnostic and biochemical features of generalised glycogenosis type II in Brahman cattle

SUMMARY Clinical, diagnostic and biochemical features of generalised glycogenosis are described In 96 Brahman-type calves. Typically the calves were presented when about 6 months of age, with III-thrift and muscular weakness as the most common signs. Acidic α-glucosidase activity was reduced in peripheral blood lymphocytes and skeletal muscle. Muscle glycogen concentration was consistently higher in affected animals than In clinically normal cattle. Other observations in affected calves Included elevation of serum aspartate amlnotransferase and creatine kinase activities and excessive amounts of high molecular weight oligosaccharides in urine. Fine cytoplasmic vacuolation of neurones In the brain and spinal cord, skeletal muscle, myocardlum and of Purkinje fibres were consistent histological observations. Periodic acid-Schlff staining revealed the presence of glycogen-like material In peripheral blood lymphocytes of all affected calves, Indicating that this is a useful aid for the diagnosis of glycogenosis. While 3 of the 96 calves showed somewhat different clinical signs, the similarity of pathology and the biochemical and clinical evidence in the remainder suggested that, In these animals, the disease was expressed as a single syndrome.     

Genotyping Shorthorn cattle for generalised glycogenosis

OBJECTIVE: To develop a procedure for routine genotyping of Shorthorn cattle for the generalised glycogenosis allele in exon 18 of the acidic alpha-glucosidase gene. PROCEDURE: Allele-specific amplification and double mismatch amplification procedures for the discrimination of the exon 18 alleles were evaluated using leucocytes and hair roots as sources of target DNA. RESULTS: Allele-specific amplification was effective for genotyping Shorthorn cattle at the 2454 site when purified DNA was used as target for the polymerase chain reaction. However, when the target DNA was derived from hair roots, differences in the relative yield of wild-type and mutant amplicons were observed. The double mismatch amplification procedure was effective in genotyping all subjects, independent of the source of DNA. The unique cleavage sites for Drd I and PshA I within exon 18 are present and absent respectively in the wildtype amplicon, and are lost and acquired, respectively, in the mutant amplicon. In addition, the Drd I and PshA I mismatching cleavage sites incorporated into the primers serve as internal controls for Drd I and PshA I cleavage. CONCLUSION: The double Drd I/PshA I mismatch amplification procedure using hair root samples as the source of DNA is a robust method for genotyping Shorthorns for generalised glycogenosis.     

Evidence of molecular heterogeneity for generalised glycogenosis between and within breeds of cattle

SUMMARY Northern analyses revealed normal levels of acidic α-glucosidase mRNA in cultured fibroblasts from a Shorthorn calf affected with glycogenosis but a gross deficiency in an affected Brahman calf. Analyses of acidic α-glucosidase activity, relative to that of other lysosomal enzymes, in blood mononuclear cells revealed greater variation within and between Brahman herds than Shorthorn herds. A Msp1 restriction fragment length polymorphism associated with glycogenosis in Brahmans was not found in Shorthorns. These results are considered in relation to molecular heterogeneity for AAG deficiency in cattle and its implications for disease control programs.     

Narcolepsy in a Brahman bull

Behavioral, electrophysiologic, and pharmacologic tests were performed on a 2 1/2-year-old Brahman bull suspected of having narcolepsy. Placement on a tilt table or electroejaculation induced apparent sleep and cataplexy. Electrophysiologic recordings during the episodes revealed the low-voltage, high-frequency electroencephalogram, reduced electromyogram amplitude, and rapid eye movements in the electrooculogram associated with rapid eye movements sleep. Physostigmine salicylate appeared to elicit and potentiate the episodes, whereas atropine sulfate and imipramine reduced or blocked them. The combined results of the tests supported the diagnosis of narcolepsy.     

Control of bovine leukosis virus in a dairy herd by a change in dehorning.

Following the demonstration that bovine leukosis virus was transmitted in calves by gouge dehorning, electrical dehorning at a younger age was implemented in a commercial Holstein herd. Subsequently, annual testing of the herd revealed a decline in the prevalence of bovine leukosis virus antibodies as older cattle dehorned by the former method were replaced by younger cattle dehorned by the latter method.     

(Co)variance components, genetic parameters and annual trends for calf weights in a pedigree Brahman herd under selection for three decades

A 300 cow Brahman herd kept on improved pasture was subjected to a selection and management programme based on a limited breeding season. Artificial insemination using mainly progeny tested bulls was used in part of the herd and the rest were bred in single sire herds. Of the 200 sires used during the 30 year period, 82% were homebred and selected principally for high estimated breeding value of 18-month weight. Variance components of birth (BW), weaning (205 W) and 18-month (548 W) weights of 6130 calves born 1968 through to 1997 were estimated by the Restricted Maximum Likelihood method (REML) using uni- and bivariate animal models. For each weight the animal's direct and maternal genetic and the dam's permanent environmental effects were considered random and those of sex, year and month of birth and age of cow were considered fixed, but the models differed as far as the number of significant interactions included. Adjusted least squares means for BW, 205 W and 548 W were 28, 158 and 292 kg. Phenotypic and direct and maternal genetic trends from univariate analysis were for BW: 0.156, 0.061 and −0.001 kg; for 205 W: 0.471, 0.126 and 0.044 kg; for 548 W: 1.973, 0.486 and 0.251 kg per year. Direct and maternal heritabilities from univariate analyses were for BW, 205 W and 548 W, 0.33 and 0.08; 0.07 and 0.14; 0.13 and 0.08, respectively. Genetic direct-maternal correlations for the three weights were −0.37, −0.13 and 0.49 and permanent environmental variance of the dam as proportion of phenotypic variance (c²) had values of 0.03, 0.16 and 0.01, respectively. Direct and maternal genetic correlations were for BW: 205 W, 0.64 and 0.74; for BW: 548 W, 0.35 and 0.74; and for 205 W: 548 W, 0.64 and 0.96. Future genetic work in the herd should put more emphasis on the improvement of cow efficiency for sustainable beef production on native and improved pasture.     

A herd level analysis of a Corynebacterium pseudotuberculosis outbreak in a dairy cattle herd

Corynebacterium pseudotuberculosis infection in an Israeli dairy cattle herd is described. The disease was characterized by ulcerative granulomatous lesions, which occurred in an epidemic form. Thirty-two cows and two heifers were affected, the ratio of the number affected to number at risk being 17.5 : 1 and 9.5 : 1, respectively. The culling rate was 50% of the affected animals. Most of the affected animals were cows (91.2%), with one first-calving cow (2.9%) and two heifers (5.9%) also affected. The infection occurred during the summer to autumn months (August-December), and lasted 118 days. The incubation period is about 2 months. The disease appeared in two clinical forms - cutaneous and mastitic - or as a mixed form. C. pseudotuberculosis organisms that were isolated from the ulcerative granulomatous lesions and from milk samples failed to reduce nitrate. A decrease in milk production (4%) and an increase in the bulk-milk somatic cell count from a herd mean of 240 x 10(3) mL(-1) to 460 x 10(3) mL(-1) were noted during the morbidity period. The organism was isolated from milk samples of eight animals (25%). Clinical, epizootiological and microbiological aspects of the infection are described.     

Congenital cataracts in an Ayrshire herd: a herd case report

Background

Microbiological standards within pork slaughter processing plants in the European Union are currently governed by Commission Regulation (EC) 2073/2005, which describes detailed performance criteria at specific stages of the procedure (following carcass dressing and before chilling) for total viable counts (TVC), Enterobacteriaceae (EB) and Salmonella spp. In this study, 95 carcasses from an Irish pork slaughter plant were sampled by swabbing 100 cm² of surface at three sites (belly, ham, jowl) to examine the effects of eight processing stages (stunning, bleeding, scalding, singeing, polishing, evisceration, final inspection and chilling) on contamination levels.

Results

TVC ranged from approximately 1.7–6.3 log cfu cm² during sampling. There were significant reductions in TVC for all sites after scalding and singeing (p < 0.05), whilst there was a significant increase in counts after polishing and evisceration (p < 0.05) compared with preceding stages. EB counts indicated hygienic weak points in the examined slaughter plant leading to faecal (cross)-contamination, with elevated counts after stunning, bleeding and evisceration (p < 0.05), compared with final counts after chilling.

Conclusions

Although the bacterial numbers reported in this study may reflect specific plant practices and temporal influences, results show that contamination can be introduced at various steps in the process and highlight the importance of monitoring locations other than those required by legislation within the process. Monitoring can be used to establish baseline levels for high-risk stages specific to each plant and to assess the effectiveness of additional interventions.     

Monensin toxicosis in a dairy herd

An acute onset of illness was reported in a lactating dairy herd in Canada in which monensin had been incorporated in error in its ration at almost 10 times the prescribed level. All of the lactating cows became lethargic, were inappetant, developed diarrhea, and had decreased milk production from 28 kg/cow/day, the day before exposure, to 23 kg/cow/day, 3 d later. Within 9 d, all animals recovered from the incident and milk production returned to previous levels.     

Cerebrospinal nematodiasis in a goat herd

During the fall of 1985, 4 Angora goats, from a herd of 40, were examined on a farm in central Michigan. Affected goats were alert but had neurologic deficits consistent with upper and lower moto neuron involvement. Eosinophilic pleocytosis in a cerebrospinal fluid sample from one goat was consistent with cerebrospinal nematodiasis. Parelaphostrongylosis was confirmed in 3 goats by identification of Parelaphostrongylosis tenuis larvae in spinal cord sections. Ivermectin may have influenced the herd's susceptibility to new parasitic infections. Control of parelaphostrongylosis probably is best achieved by removal of susceptible animals from treed swamps coinhabited by white-tailed deer from late summer until after the first killing frost.     

Phosphorus deficiency in a dairy herd

Phosphorus deficiency was diagnosed in a 90 cow seasonal supply dairy herd which showed low milk production, ill-thrift, infertility and osteophagia. Serum inorganic phosphorus and pasture phosphorus levels were low, pasture Ca:P ratios high and soil phosphorus levels very low; the soil phosphate retention value was 96%. The deficiency, which had probably existed for several years, was considered to be due to inadequate annual phosphate fertiliser applications on a high phosphate retaining soil. Phosphorus deficiency may have been exacerbated by the application of lime without phosphate several months before the onset of severe clinical disease. The finding of low sodium levels in pasture samples suggested that sodium deficiency may have co-existed and contributed to the clinical picture. Treatment, comprising bone flour dusting of pasture, water trough supplementation and phosphorus-containing injections, appeared to induce ovarian activity in anoestrous cows and suppress osteophagia, though controlled treatment trials were not performed. No improvement was noted in milk production or cow condition. Increased annual super-phosphate and reduced potassium applications were recommended, together with the monitoring of pasture and soil macro elements and serum phosphorus levels. A decision on whether to supplement with sodium would be based on the results of pasture monitoring, since a direct animal test is not currently available. It is suggested that phosphorus deficiency may not be uncommon in dairy herds in some North Island districts; based on the present case, the decline in New Zealand superphosphate quality in the 1970's and recently published data which has shown a high proportion of pasture phosphorus deficient sites in some North Island areas.     

Paratuberculosis in a large goat herd

Paratuberculosis was studied for three years in a large goat herd. Post mortem lesions were seen mainly in the ileum and mesenteric lymph nodes where caseation was often present. In three successive years the annual faecal excretor rate was 15, 9 and 13 per cent despite removal of positives. Faecal culture and an agar gel immunodiffusion test proved useful for detecting the disease in live goats, but complement fixation and tuberculin tests were of less value. The prospects of controlling the disease are discussed.