Maple syrup urine disease in five hereford calves in ontario

Five newborn grade horned or polled Hereford calves in Ontario exhibited the clinical signs characteristic of hereditary neuraxial edema, namely, inability to rise, and hyperesthesia with extensor spasms upon tactile or auditory stimulation. Histologically, there was widespread spongy vacuolation in both white and gray matter in the brains of all five calves, especially in the midbrain, brain stem, and cerebellum. Amino acid analysis of formalin-fixed cerebral tissue of three calves demonstrated an amino acid composition consistent with a diagnosis of branched chain keto acid decarboxylase deficiency. This condition has been recently reported in Hereford calves in Australia; it is characterized by the delayed onset of neurological signs and severe lesions of status spongiosus, and has been considered analogous to maple syrup urine disease of children.     

Maple syrup urine disease in calves: a clinical, pathological and biochemical study

The clinical, pathological and biochemical findings of a study of 30 Poll Hereford, Hereford, Poll Hereford cross or Hereford cross calves affected with branched chain ketoacid dehydrogenase (BCKAD) complex deficiency or maple syrup urine disease (MSUD) are presented. In breeding studies, 6 of 21 calves from obligate heterozygote matings were affected with MSUD, suggesting the disease is inherited in an autosomal recessive manner. Calves were clinically affected from birth, but there were variations in the subsequent course of progressive deterioration of central nervous system function. Concentrations of the branched chain amino acids and keto acids were elevated in pre-suckle plasma and cerebellar water content was higher in affected calves. Activity of BCKAD complex was minimal in fibroblasts cultured from an affected calf. Spongiform encephalopathy and elevated ratios of the branched to straight chain amino acids in formalin fixed cerebral tissue were found in a stillborn foetus and a 3-month-old Hereford calf. These findings suggest the disease occurs prenatally and that a delayed form may exist.     

Definition of the mutation responsible for maple syrup urine disease in Poll Shorthorns and genotyping Poll Shorthorns and Poll Herefords for maple syrup urine disease alleles.

The organisation of the E1alpha subunit of bovine branched-chain alpha-keto acid dehydrogenase gene was established. c DNA was cloned from Poll Shorthorn x Poll Hereford calves affected with Maple Syrup Urine Disease to identify the mutation responsible for the disease in Poll Shorthorns. Clones containing the c DNA sequences inherited from the Poll Shorthorn sire of the affected calves were identified. Paternal clones were sequenced and a cytidine to thymidine transition was found at nucleotide 1380. The mutation is predicted to substitute leucine in place of a highly conserved proline at codon 372. A polymerase chain reaction procedure was developed for detection of the 1380C-->T mutation in genomic DNA. Three Poll Shorthorn parents of affected calves and three affected Poll Shorthorn x Poll Hereford calves were heterozygous and an affected Poll Shorthorn calf was homozygous for this mutation. An improved polymerase chain reaction procedure was also devised to genotype Poll Herefords for the 248C-->T mutation. The procedures will facilitate disease prevention programs and assist in differential diagnosis of conditions in new-born calves that present with a rapid onset of progressive neurological disease and are characterised histologically by 'status spongiosus'. Maple Syrup Urine Disease (MSUD) is an autosomal recessive defect reported in humans (Danner and Elsas 1989), and in Poll Hereford (PH) and Poll Shorthorn (PS) calves (Harper et al 1986, Healy et al 1992). The clinical, biochemical and pathological manifestations of the disease are identical in the two breeds of cattle, and are characterised by the rapid onset of progressive neurological disease, leading to death within a few days of birth. The disease is caused by a deficiency of activity of the mitochondrial enzyme branched-chain alpha-keto acid dehydrogenase (BCKADH). This deficiency leads to elevated concentrations, in blood and tissues, of branched chain alpha-keto acids and their precursors, the branched chain amino acids, valine, leucine and isoleucine. BCKADH consists of four subunits E1alpha, E1beta, E2 and E3 that are encoded by separate genes, and MSUD may result from deficiency of any of the subunits. In PH s, the disease in caused by premature termination of translation, of the E1alpha subunit, that is induced by a cytidine to thymidine transition exon 2 (248C-->T), that converts the glutamine codon -6 to a stop codon (Q-6ST; Zhang et al 1990). We have shown that MSUD -affected PSxPH calves are heterozygous at the PH locus, illustrating molecular heterogeneity exists for bovine MSUD (Healy and Dennis 1994a). The fact that these crossbred calves are affected, indicates the PS, like the PH mutation, resides in the E1alpha subunit.     

D-lactate production and excretion in diarrheic calves

The origin of D-lactate, the most important acid contributing to metabolic acidosis in the diarrheic calf, is unknown. We hypothesized that because D-lactate is produced only by microbes, gastrointestinal fermentation is the source. The objective of this study was to determine whether D-lactate production occurs in the rumen, colon, or both, and to measure D- and L-lactate concentrations in urine. Fecal, rumen, blood, and urine samples were obtained from 16 diarrheic and 11 healthy calves. Serum electrolyte concentrations were measured in both groups, and blood gas analyses were performed for diarrheic calves. All samples were analyzed for D- and L-lactate by high performance liquid chromatography (HPLC). Diarrheic calves were generally hyperkalemic with high serum anion gap, depressed serum bicarbonate, and low blood pH. L-lactate was markedly higher in rumen contents (22.7 mmol/ L [median]) and feces (8.6 mmol/L) of diarrheic calves than healthy calves (0.5 mmol/L and 5.1 mmol/L, respectively), but not different in serum or urine. Rumen, fecal, serum, and urine D-lactate concentrations were all significantly higher (P < .05) in diarrheic calves (17.0, 25.4, 13.9, and 19.2 mmol/L, respectively) than in healthy calves (0.5, 9.1, 1.4, and 0.5 mmol/L, respectively). Higher D-lactate concentrations in the rumen and feces of diarrheic calves suggests these sites as the source of D-lactate in blood and urine.     

Heterozygote detection for maple syrup urine disease in cattle

SUMMARY The clinical, pathological and biochemical manifestations of maple syrup urine disease (MSUD) are similar in Poll Hereford and Poll Shorthorn X Poll Hereford calves. No significant differences were observed in branched-chain amino acid concentrations in plasma, or of branched-chain keto acid dehydrogenase activity in fibroblasts, between Poll Herefords homozygous normal and heterozygous for the mutation responsible for MSUD. Haemopoietic chimerism resulted in incorrect diagnosis of the MSUD genotype in 30% of non-identical twins when blood DNA was analysed using allele-specific amplification. Hair roots are shown to be a suitable source of target DNA for genotyping Poll Hereford cattle for the MSUD mutation. Twelve of 203 (5.8%) aged Poll Hereford bulls, sampled at saleyards during the last 4 months of 1993, were found to be heterozygous for the mutation. In contrast, the mutant sequence was detected in only 1 of 150 (0.7%) 2- and 3-year-old Poll Hereford bulls offered for sale at 2 stud sales held during 1993, suggesting that the prevalence of the disease may decline over the next few years.     

Myophosphorylase deficiency (glycogen storage disease Type V) in a herd of Charolais cattle in New Zealand: confirmation by PCR-RFLP testing

AIM: To describe a disease of muscle in Charolais calves and confirm the putative diagnosis of inherited myophosphorylase deficiency. METHODS: Variously stained paraffin sections of muscle prepared from affected calves were used to describe the lesions. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) test was developed and applied to affected calves, their sires, dams and other individuals. RESULTS: The lesions were those of rhabdomyolysis of skeletal muscles and sub-sarcolemmal spaces in normal fibres. The PCRRFLP test confirmed the expected mutation for phosphorylase deficiency of Charolais cattle in two affected calves. In addition, sires, dams and other closely-related individuals of four affected calves tested as heterozygous for the mutation. Other apparently unrelated animals also tested as heterozygous. CONCLUSIONS: The diagnosis of myophosphorylase deficiency was confirmed. The PCR-RFLP test is suitable for use in controlling this recessively-inherited disorder as it can diagnose heterozygous individuals that are otherwise clinically normal.     

Myophosphorylase deficiency (glycogen storage disease Type V) in a herd of Charolais cattle in New Zealand: Confirmation by PCR-RFLP testing

AIM: To describe a disease of muscle in Charolais calves and confirm the putative diagnosis of inherited myophosphorylase deficiency.

METHODS: Variously stained paraffin sections of muscle prepared from affected calves were used to describe the lesions. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) test was developed and applied to affected calves, their sires, dams and other individuals.

RESULTS: The lesions were those of rhabdomyolysis of skeletal muscles and sub-sarcolemmal spaces in normal fibres. The PCR-RFLP test confirmed the expected mutation for phosphorylase deficiency of Charolais cattle in two affected calves. In addition, sires, dams and other closely-related individuals of four affected calves tested as heterozygous for the mutation. Other apparently unrelated animals also tested as heterozygous.

CONCLUSIONS: The diagnosis of myophosphorylase deficiency was confirmed. The PCR-RFLP test is suitable for use in controlling this recessively-inherited disorder as it can diagnose heterozygous individuals that are otherwise clinically normal.     

铜缺乏对奶牛红细胞及组织细胞膜ATP酶活性的影响

为探讨铜缺乏对奶牛红细胞及组织细胞膜ATP活性的影响，在调查铜缺乏奶牛水土食物链微量元素状态的基础上，首次测度了病区6头发病犊奶牛（Ⅱ组）及该地区20头同年生临床健康奶牛（IH组，血清铜低于0.56mg/kg）红细胞膜Na^ -K^ -ATP酶、Ca^2 -ATP酶及Mg^2 -ATP酶的活性，对20头IH组奶牛进行为期80d的补铜试验（含铜40mg/kg干饲料），以20头健区同年生健康奶犊牛作为对照（HH组，全血铜大于０.85mg/L）。同时测试了剖杀后６头病牛及健康牛组织细胞膜ＡＴＰ酶的活性。结果表明：Ⅱ组奶牛红细胞膜ＡＴＰ酶活性显著低于ＨＨ组及ＩＨ组。补铜后４０d，ＩＨ组奶牛红细胞膜Na^ -K^ -ATP酶均显著升高，病牛肝、脾、肾、大脑、小脑、淋巴结等组织细胞膜Na^ -K^ -ATP酶、Ca^2 -ATP酶活性显著低于对照组。结论：铜缺乏较为严重地影响了奶牛红细胞及组织膜ＡＴＰ酶的活性，抑制了这些组织的正常生理功能。     

Stillbirth/perinatal weak calf syndrome in dairy heifers: results of a pilot study

A pilot study of 10 Dutch dairy herds was performed to investigate possible causes of stillbirth/perinatal weak calf syndrome in heifers. Possible causes and advice for further investigations on farms affected by this syndrome are discussed based on the results of investigations of blood and urine from pregnant young stock, postmortem examination of still-born calves, and a questionnaire held among farmers. Infections with Neospora caninum, Salmonella spp. or Leptospira hardjo, or a deficiency of iodine could be excluded.     

Status spongiosus of white matter in newborn Gelbvieh-cross calves.

Various forms of status spongiosus occur in neonatal cattle, the best characterized of which is due to mutations of the branched-chain alpha-keto acid dehydrogenase complex (BCKD), resulting in bovine maple syrup urine disease (MSUD, branched-chain ketoaciduria). A distinctive neurological syndrome was identified between 1998 and 2003 in 9 calves in a 250-cow stabilized Gelbvieh-Red Angus herd. Both sexes were affected (6 heifers, 3 bulls), with a low annual incidence (3 cases in 1998; no cases in 1999; 2 cases in 2000; 2 in 2001; 1 in 2002; 1 in 2003). Affected calves were born full-term, unable to stand, and had constant whole-body tremors when stimulated. Animals remained in lateral recumbency until death or euthanasia; the longest survival time was 10 days postpartum. The principal histological change in 2 affected calves was diffuse, moderately severe bilaterally symmetrical status spongiosus with Alzheimer type II cells throughout the white matter of the brain. Myelin deficits were not evident and vacuoles were due to cleaved myelin sheaths. Neither recognized mutation of MSUD was identified in the E1a subunit of BCKD in 2 affected calves, 8 dams that gave birth to affected calves, a grand-dam of 3 affected calves, or a sire of 1 calf. Amino acid analysis of serum from 1 affected calf revealed normal concentrations of branched-chain amino acids, indicating that this disease is distinct from MSUD. The genetic and biochemical basis for the disorder, provisionally named congenital status spongiosus of Gelbvieh-cross cattle, is undetermined. The pattern of inheritance was not established.     

Mucosal uptake,mucosal transfer and retention of iron in veal calves

A method for studying iron absorption in humans was adapted to veal calves. Three 10-week-old calves with moderate (calves 1 and 2) or severe (calf 3) iron deficiency were given an abomasal injection of⁵⁹Fe and⁵¹Cr and all their faeces were collected over 15 days in order to measure mucosal uptake, mucosal transfer and retention of iron. The mucosal uptake was 62.2, 53.4 and 71.8% in calves 1, 2 and 3, respectively. The iron retention measured 14 days after administration of the test dose was 57.4, 52.3 and 56.4% in calves 1, 2 and 3, respectively. Maximal plasma activity was found in all three calves between 1 1/2 and 2 h after injection of the test dose. The plasma activity decreased rapidly, with a slight increase between the 5th and the 10th hour. After 21 h, less than 0.25% of the injected dose was still present in 1 litre of plasma. Not all the⁵¹Cr was recovered in the faeces. No⁵⁹Fe was found in the urine but some⁵¹Cr could be detected. The results of this study show that the method described is useful for measuring the different steps of iron absorption in iron-deficient veal calves.Abbreviations Hb haemoglobin concentration - MCV mean corpuscular volume - PCV packed cell volume - PI plasma iron - RBC red blood cell count - R iron retention - TIBC total iron-binding capacity - TIBC-SAT saturation of the total iron-binding capacity - U mucosal uptake     

Use of furosemide to obtain bovine urine samples for leptospiral isolation

The use of the diuretic furosemide made it possible to obtain samples of urine from cattle for leptospiral isolations. The drug was injected IV at a dose level of 0.8 mg/kg for heifers and 0.5 mg/kg for calves. The average time to first voiding in heifers was 19 minutes. The average time from the first to the second voiding was 17 minutes. The average time to the first voiding in four calves was 12 minutes; the average time from the first to the second voiding was 10 minutes. A decrease in urinary osmolarity provoked by furosemide created a more favorable condition for the survival of leptospires. Leptospires were isolated in 24 (72.7%) of 33 weekly cultural attempts with the aid of furosemide in three experimentally infected adult cattle. Serovar hardjo was isolated in 16 (57.1%) of 28 weekly cultural attempts with aid of the diuretic in four experimentally infected calves. The recovery frequency was 28.5% from the first voiding and 50% from the second. Leptospires were not isolated from urine obtained from the calves by manual stimulation. Untoward side effects that might have been attributable to furosemide were not observed. Furosemide appears to be well suited to obtain urine samples from cattle for leptospiral isolation.     

Clinical and biochemical alterations in calves with nutritionally induced polioencephalomalacia

Polioencephalomalacia (PEM) was induced in calves by feeding a semipurified, low-roughage diet of variable copper and molybdenum composition. Two formulations resulting in Cu-insufficient and Cu-sufficient forms of the diet were fed (n = 10 and 4 calves, respectively); both diets induced PEM. Clinical signs of disease developed as early as 15 days after transition to the experimental diets and included impaired vision, decreased response to external stimuli, and abnormal gait. Grossly evident cerebrocortical lesions consisted of laminar areas of cavitation and/or autofluorescence seen under UV illumination. Hepatic Cu concentration was decreased in calves fed the Cu-insufficient diet, but not below normal range. During the course of feeding either diet, rumen pH decreased, rumen volatile fatty acid concentrations increased, rumen and blood lactic acid concentrations increased, and rumen and plasma thiamine concentrations increased. The thiamine pyrophosphate effect on erythrocyte transketolase activity was unaltered in calves of either diet group. This nutritionally induced form of PEM does not appear to be related to Cu deficiency or reduction in plasma or rumen thiamine concentration.     

Experimental infection of calves with Leptospira interrogans serovar hardjo: conjunctival versus intravenous route of exposure

Eight-month-old calves, housed under maximum isolation, were exposed to pathogenic Leptospira interrogans serovar hardjo by the conjunctival route or IV. One calf served as an unexposed control. Infection was monitored serologically (microscopic agglutination test and enzyme-linked immunosorbent assay; ELISA) and by leptospiral culture isolation from periodic urine samples and from the kidneys, epididymides, and aqueous humor collected at slaughter. Microscopic agglutination test titers of greater than or equal to 1:40 were detected among all IV exposed calves at postinoculation day (PID) 7 and among conjunctival exposed calves at PID 14. By ELISA, all IV exposed calves were positive by PID 3, whereas conjunctival exposed calves were positive at PID 14. The ELISA was more sensitive for the detection of antibodies against leptospires in cattle. Leptospires were isolated from the urine of 4 calves and from the kidney of 3 calves exposed by the conjunctival route, but not from IV exposed calves. The results indicated that the conjunctival route of exposure was a more natural and successful route for experimental infection of cattle with serovar hardjo.     

Kyphosis in Jersey calves

A condition of Jersey calves involving kyphosis and lordosis is described. Four calves originating from three farms, all the progeny of one bull, were necropsied. The only significant finding was a myopathy mainly affecting sublumbar muscles. The lesions were not consistent with white muscle disease or of any known myopathy affecting Jersey calves. It is likely that this condition represents a primary muscular dystrophy of genetic origin.     

Experimental infection of calves with an adenovirus isolated from sheep and related to bovine Adenovirus type 2 I. Clinical and virological studies.

Colostrum deprived calves were experimentally infected with an adenovirus isolated from sheep and related to bovine adenovirus type 2. The calves showed respiratory symptoms and mild diarrhoea from the third day after infection. Laboratory tests revealed the development of leucopenia, lymphopenia, a drop of the pH of the urine and the appearance of pathological changes in the urine. The animals shed the virus in their nasal discharge, faeces and urine. Comparing the clinical and virological findings with the previous experimental infection of lambs it is concluded, that this type of adenovirus is similarly pathogenic for the two ruminant species.     

铜缺乏奶牛琥珀酸脱氢酶的组化特征

为探讨铜缺乏对奶牛心脏、肝脏、肾脏琥珀酸脱氢酶 (SDHase)组化特征 (分布特点及活性 )的影响 ,本实验选择铜缺乏症高发区自然发病的奶犊牛 6头 ,剖杀后采集心、肝、肾作冰冻切片及酶组化染色 ,以 6头同年生健康奶犊牛为对照。实验结果表明 ,发病组奶牛心脏中 SDHase计数值显著高于对照组 (P<0 .0 5 ) ,而肝脏、肾脏中SDHase计数值显著低于对照组 (P<0 .0 5 ) ;健康组奶牛心脏、肝脏、肾脏中 SDH活性扫描值均显著大于发病组(P<0 .0 5 )。发病奶牛组组织中 SDHase酶颗粒的分布也发生特征性的变化。结论 :氧化呼吸链关键酶活性的改变是导致奶牛铜缺乏症病理过程的重要原因     

Modification of the concentration of creatinine and sodium in blood serum and urine and the fractional elimination of sodium by electrolyte therapy in calves]

In 32 calves, creatinine and sodium concentrations were determined in blood and urine samples taken simultaneously four times during two periods of two days each (control period: milk or milk replacer; trial period: oral electrolyte solution). Creatinine urine/serum ratios and fractional elimination of sodium (FENa) were calculated. The influence of the type of liquid feed on these parameters was evaluated statistically. Calves fed milk replacer had higher urine sodium concentrations and FENa than calves fed whole milk. All parameters determined were influenced by intake of oral electrolyte solution: creatinine concentrations in serum and urine decreased, whereas sodium concentrations in serum and urine, as well as FENa, increased. It is concluded that sodium content of liquid feed significantly influences FENa.     

Hematology of the neonatal calf. III. Frequency of congenital iron deficiency anemia.

The hematocrits of 258 Holstein-Friesian and Jersey calves were determined during the first 3 days following birth. Hematocrit values of 25% or less were observed in 15.8% of the calves and hematocrits of 20% or less were observed in 6.9% of calves from apparently normal dams. The serum iron concentration of neonatal calves with hematocrits of 25% or less was significantly lower than the value recorded in calves with hematocrits above 25% and there was a direct correlation between PCV and serum iron concentration. The data presented suggested that prenatal development of iron deficiency is an important cause of anemia in neonatal dairy calves. Factors which may be important in pathogenesis are discussed.