A study of nutritional myopathy in weaner sheep

The effectiveness of various treatments upon, and pathological and biochemical changes in, ovine weaner nutritional myopathy were observed. Clinical myopathy was already apparent in the sheep at the start of the study, and they were fed decreasing amounts of a ration containing low levels of selenium and alpha-tocopherol, and periodically deprived of water. In spite of this management there was a spontaneous remission of the clinical myopathy in the sheep, but a subclinical myopathy was identified in some of the sheep at the end of the trail. The conclusions were that the myopathy was not caused by a low dietary intake of selenium and/or alpha-tocopherol alone, that alpha-tocopherol was involved in the aetiology, that alpha-tocopherol was completely effective and selenium possibly partially effective in treating it, and that the condition may be a Type II muscle fibre disease. Data on plasma creatine phosphokinase and erythrocyte glutathione peroxidase activities, and terminal liver selenium and alpha-tocopherol concentrations are presented, and their roles in the diagnosis of ovine weaner nutritional myopathy discussed.     

Muscle enzymes in the diagnosis of ovine weaner nutritional myopathy

SUMMARY The diagnostic performance of plasma tests for muscle enzymes was measured in sheep from flocks affected by clinical and sub-clinical nutritional myopathy. Parallel combinations of tests for creatine kinase (CK), alanine amino transferase (ALT), aspartate amino transferase (AST) and lactate dehydrogenase had higher diagnostic sensitivity than CK alone. The enzymes ALT and AST showed the highest correlation with the degree of muscle damage. A parallel combination of tests for plasma CK and ALT as well as tests for plasma α-tocopherol and red cell glutathione peroxidase are recommended for the diagnosis of nutritional myopathy and a decision on the appropriate treatment. The number of false negative results based on a diagnosis from the microscopic examination of single muscles was higher than for the parallel combination of tests. The number of false negatives was highest for the vastus intermedius and lowest for the tensor fascia lata. Diagnosis using a panel of blood tests has the advantages of overcoming problems of inadequate muscle sampling, a larger number of sheep in the flock can be tested and a more rapid diagnosis can be obtained.     

Plasma and liver concentrations of aP-tocopherol in weaner sheep after vitamin E supplementation

Changes in the plasma and liver concentrations of alpha-tocopherol in sheep kept in an animal house, grazed on pasture, grazed on pasture and fed an oat/lupin grain supplement, or grazed on toxic lupin stubble were monitored following no, oral or intramuscular administration of alpha-tocopherol. It was concluded that intramuscular administration was most effective because it caused longer periods of elevated plasma alpha-tocopherol concentrations in most situations investigated and consistently higher concentrations of alpha-tocopherol in the liver, compared to oral administration. The dose rates of alpha-tocopherol given, the vehicle for alpha-tocopherol in the injectable preparation, the different diets consumed and the concurrent occurrence of lupinosis all influenced the results. In spite of this a significant positive correlation existed between the terminal liver and plasma alpha-tocopherol concentrations.     

A nutritional myopathy enzootic in a group of yearling beef cattle

Steers and bulls under feedlot conditions and on an apparently adequate ration developed transitory signs of diarrhea and unthriftiness. One animal became recumbent. Sick and clinically normal animals in the group had elevated serum creatine kinase and aspartate aminotransferase enzyme values and degenerative changes in muscle fibers on biopsy. Feed analysis was carried out and the animals were monitored over several weeks by means of muscle biopsy and serum enzyme analysis as well as by postmortem examination at slaughter. Subclinical myopathy was found in several animals and was thought to be related to low vitamin E content in the high moisture corn ration.     

Fibrotic myopathy of the semitendinosus muscle in a cat

Fibrotic myopathy of the semitendinosus muscle causing a progressive mechanical right hind limb lameness in a cat was treated with a Z-plasty lengthening procedure of the affected muscle. Lameness resolved after surgery; however, recurrence of gait abnormality began 2 weeks later. One year after surgery, the cat's gait was abnormal; this had little effect on ambulation. Histologic examination of biopsy specimens taken from the affected muscle at surgery revealed complete replacement of myofibers with dense fibrous connective tissue. Etiopathogenesis was not determined.     

Fibrotic myopathy of the iliopsoas muscle in a dog

Fibrotic myopathy of the iliopsoas muscle developed in a dog, following extensive migration of a grass awn within the muscle and adjacent subcutaneous tissue. The dog was initially presented for evaluation of a fluctuant swelling over the right flank region. The clinical and imaging findings were suggestive of iliopsoas fibrotic myopathy and the diagnosis was confirmed by histopathology. This is the third report of iliopsoas fibrotic myopathy in a dog, the first report to describe the postmortem pathologic changes and the first report of iliopsoas fibrotic myopathy subsequent to foreign body migration.     

The effect of thiaminase-induced subclinical thiamine deficiency on growth of weaner sheep

Three experiments were performed to examine for causes of poor growth of young Merino sheep. Weekly testing of animals 42 weeks of age for 10 weeks revealed that 90% of clinically poor animals were excreting high levels of thiaminase in their faeces; low levels of activity were present in 20% of clinically normal animals. There were significant differences in the mean erythrocyte transketolase activity of the thiaminase excreting poor animals and the thiaminase free normal animals. Other known causes of poor growth could not be demonstrated. Weekly monitoring of thiaminase activity in the faeces from 80 lambs 6 weeks of age showed 23% to be excreting significant levels of enzyme (>3mUg^–1 DM) throughtut a 10 week test period. Mean growth rates of these lambs were significantly below those of lambs not excreting thiaminase or excreting low levels intermittently. Supplementation of thiaminase excreting lambs with intra-muscular injections of thiamine HCl was associated with a statistically significant improved growth rate (P<0.01) compared to unsupplemented sheep excreting thiaminase. Mean growth rates of lambs not excreting thiaminase on a continuous basis (sampled weekly) were the same with or without thiamine HCl supplementation. High thiaminase levels were found in the ruminal fluids of trial animals excreting the enzyme in their faeces, confirming this previously established association.Bacillus thiaminolyticus was isolated from faeces and ruminal fluids from clinically poor animals and is the most likely source of the thiaminase. Subclinical thiamine deficiency was indicated by low erythrocyte transketolase activities and elevated TPP effects and is proposed as the cause of the poor growth by the young sheep.     

Experimental myopathy in vitamin E- and selenium-depleted calves with and without added dietary polyunsaturated fatty acids as a model for nutritional degenerative myopathy in ruminant cattle

Fourteen calves, 14 weeks old, fed a low vitamin E and selenium diet based on sodium hydroxide-treated, selenium-deficient barley showed a rapid decline in plasma alpha-tocopherol, a gradual decline in erythrocyte glutathione peroxidase (GSHPX) activity, elevated plasma creatine kinase (CK) activity, teeth grinding and (in three cases) electrocardiographic changes, but no clinical abnormalities after 127 to 137 days. Necropsy of six depleted calves at that stage revealed pale muscles in two animals and mild histopathological lesions of skeletal myopathy in all six calves. Ten control calves were fed a similar diet supplemented with alpha-tocopherol and selenium. Plasma alpha-tocopherol and erythrocyte GSHPX activity remained high in these calves and no clinical abnormalities or evidence of myodegeneration were observed. When protected linseed oil was fed as a source of polyunsaturated fatty acids to the remaining eight depleted calves, rapid elevations in plasma CK activity and linolenic acid associated with a variety of cardiopulmonary and locomotory signs, electrocardiographic changes and myoglobinuria occurred within six to 11 days. Necropsy revealed widespread severe skeletal myodegeneration (eight calves) and myocardial lesions (five calves) with preferential involvement of the left ventricular myocardium. Clinical signs and pathological changes were similar to those reported in field outbreaks of nutritional degenerative myopathy in ruminant cattle and establish the experimental regime as the first successful model for reproduction of this disease.     

The effect of thiaminase-induced subclinical thiamine deficiency on growth of weaner sheep

Three experiments were performed to examine for causes of poor growth of young Merino sheep. Weekly testing of animals 42 weeks of age for 10 weeks revealed that 90% of clinically poor animals were excreting high levels of thiaminase in their faeces; low levels of activity were present in 20% of clinically normal animals. There were significant differences in the mean erythrocyte transketolase activity of the thiaminase excreting poor animals and the thiaminase free normal animals. Other known causes of poor growth could not be demonstrated. Weekly monitoring of thiaminase activity in the faeces from 80 lambs 6 weeks of age showed 23% to be excreting significant levels of enzyme (greater than 3mUg-1 DM) throughout a 10 week test period. Mean growth rates of these lambs were significantly below those of lambs not excreting thiaminase or excreting low levels intermittently. Supplementation of thiaminase excreting lambs with intra-muscular injections of thiamine HCl was associated with a statistically significant improved growth rate (P less than 0.01) compared to unsupplemented sheep excreting thiaminase. Mean growth rates of lambs not excreting thiaminase on a continuous basis (sampled weekly) were the same with or without thiamine HCl supplementation. High thiaminase levels were found in the ruminal fluids of trial animals excreting the enzyme in their faeces, confirming this previously established association. Bacillus thiaminolyticus was isolated from faeces and ruminal fluids from clinically poor animals and is the most likely source of the thiaminase. Subclinical thiamine deficiency was indicated by low erythrocyte transketolase activities and elevated TPP effects and is proposed as the cause of the poor growth by the young sheep.     

The prevalence of Eperythrozoon ovis infection in weaner and adult sheep in north eastern Victoria

A survey of weaner (6 to 12 months) and adult sheep for the presence of Eperythrozoon ovis antibodies using an immunofluorescent antibody assay was carried out. In 22 shires in north eastern Victoria over 2 years infection was demonstrated in 10% and 51% of weaner and adult sheep respectively.     

Experimental infection of weaner sheep with S strain Mycobacterium avium subsp. paratuberculosis

We sought to determine whether infection of recently weaned 12-16-week-old Merino lambs with an Australian S strain M. a. paratuberculosis, at doses consistent with natural exposure, could be detected in the first few months post-inoculation. Such detection would facilitate the use of weaner sheep as sentinel animals for the presence of infectious doses of M. a. paratuberculosis on pastures. In controlled pen trials, oral doses of approximately 10(7)-10(8) viable organisms were demonstrated to be infective, whereas doses below 10(4) organisms failed to produce detectable infection. Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) was isolated from intestinal and/or lymphoid tissues collected at necropsy 7 or 14 weeks after first infection, but there were no associated gross or microscopic lesions. Skin testing with intradermal Johnin detected all three infected lambs at 13 weeks post-infection, and one of the three infected lambs at 6 weeks post-infection, with 100% specificity. Results for whole blood IFN-gamma assay showed some correlation with infection status but lacked specificity. One infected lamb gave a positive result in an ELISA for antibodies to M. a. paratuberculosis, 14 weeks post-infection and 1 week after skin testing. This was the first demonstration of experimental infection with S strain M. a. paratuberculosis in Australian Merino sheep at doses likely to be representative of natural infection. Culture from tissues in the first few months post-exposure could facilitate the use of naive weaner sheep as tracer animals to detect heavy contamination of pastures with M. a. paratuberculosis, but low-level contamination may not be detected in such a system.     

Plasma IGF-I binding proteins in sheep: effect of recombinant growth hormone treatment and nutritional status.

In humans the IGF binding proteins (BP) are closely related to metabolic status. In this paper we have examined the influence of controlled feed intake and GH treatment on IGF binding proteins in growing lambs. Analyses were performed on plasma samples from animals maintained on two levels of feed intake (1.75% body weight as lucerne pellets or 3% body weight which is approximately equivalent to an ad libitum intake) either with or without recombinant bovine growth hormone (BST; 0.25 mg/kg body weight/day) administration. Samples used for the analyses reported in this paper were collected at 9.00 hr following 41 d of treatment. Total plasma IGF-I was increased on the higher plane of nutrition (P less than .01) and by BST (P less than .001) but only on high feed intake. IGF is associated with BP of 150 kDa and 40-50 kDa in sheep plasma. 150 kDa bound IGF-I was increased on the higher plane of nutrition (P less than .05) and by BST treatment (P less than .001) but only on the higher feed intake. By contrast no change in 40-50 kDa bound IGF-I was observed with treatment. Unbound IGF-I was also found in sheep plasma (2-5% of total) but demonstrated only minor changes in relation to treatment. Saturation analysis gave estimates of total binding capacity and saturation of the IGF-BP. In ovine plasma the binding capacity of the 150 kDa species is in excess of bound IGF (P less than .001). Saturation did not change with treatment despite the observed differences in 150 kDa bound IGF-I. Thus BP(s) contained in the 150 kDa fraction were responsive to treatment. By contrast large differences in saturation of the 40-50 kDa species were observed (P less than .001) despite little treatment dependent change in bound IGF-I. IGF-BP(s) in the 40-50 kDa fraction were elevated in the low nutrition group and suppressed on the higher feed intake resulting in near saturation. These data strongly suggest that the IGF BP are modulated according to metabolic status in the sheep.     

Reliability of indicators of sheep welfare assessed by a group observation method

Assessments of animal-based outcomes form the core of routine veterinary clinical examinations and are being increasingly used as indicators of animal welfare. A method of group observation that did not require gathering and handling of individual sheep, was used to assess eight animal-based indicators of sheep welfare: demeanour, skin irritation, wool loss, excessive panting, coughing, lameness, and cleanliness of the ventral abdominal and 'breech' (perineum/gluteal/caudal hindlimb) areas. The inter-observer reliability of two or three observers who independently assessed these indicators was tested on 2406 adult sheep and growing lambs across 36 farms and the intra-observer reliability of an experienced, veterinary assessor--the 'test standard observer'--was assessed on 88 adult sheep during four on-farm assessments. Observer reliability was evaluated using Cronbach's alpha (α) comparison of the recorded proportion (%) of sheep affected by each welfare condition and by binomial logistic regression modelling. High levels of inter-observer reliability were identified for the assessment of group lameness (α 0.76-1.00) and cleanliness of the breech area (α 0.97-1.00). Excellent intra-observer reliability was determined for lameness (α 0.99), cleanliness of the breech area (α 0.97), demeanour (α 1.00) and wool loss (α 1.00). In addition, proportion data and logistic regression models identified few between-observer differences. The results suggest that welfare outcomes based on observations of the behaviour and physical appearance of individual animals within a group may offer a reliable and feasible measurement tool for the on-farm assessment of sheep welfare.     

Serum gamma-glutamyltransferase as a measure of sporidesmin-induced liver damage in sheep

The relationship between serum concentrations of the enzyme gamma-glutamyltransferase (GGT) after experimental sporidesmin intoxication, and the severity of the liver damage seen on post-mortem examination of the liver, has been examined. Serum GGT activity in blood collected 2 to 3 weeks after ad-ministering the toxin is positively correlated to a subjective liver damage score and to losses in bodyweight associated with the intoxication. Determination of GGT activities provides a sensitive method for detecting and measuring liver damage in experimental sporidesmin poisoning. In the field, GGT determinations should prove useful in diagnosing liver damage associated with subclinical and clinical facial eczema giving additional information about the severity of the lesion.     

Erythrocyte selenium-75 uptake as a measure of selenium status in weaner sheep, and its relationship to erythrocyte glutathione peroxidase activity

The relationship between in vitro erythrocyte 75Se uptake (75Se uptake) and erythrocyte glutathione peroxidase (EGSHPx) activity was examined in weaner sheep during periods of selenium depletion and repletion, to determine whether 75Se uptake was better correlated than EGSHPx activity to the development of weaner nutritional myopathy. In the 2 trials conducted, only 3 of 45 Merino wether weaners developed clinical myopathy and histological lesions in skeletal muscles. The 75Se uptake values and EGSHPx activities in these 3 sheep were no different from those in the unaffected sheep. There was a significant negative correlation between 75Se uptake values and EGSHPx activities over the entire period of the trials. It could not be demonstrated that 75Se uptake was any better correlated than EGSHPx activity to the development of nutritional myopathy, and it was concluded that EGSHPx activity indicated selenium status better than 75Se uptake in weaner sheep.