Cerebro-cortical necrosis (C.C.N.) in calf. An experimental reproduction of the disease

In four ruminant calves a state of thiamine deficiency was brought about with a thiamine antagonist, amprolium. Twice daily the calves were given Amprolmix® containing 25 % amprolium through a stomach tube. The dosage was 350–500 mg/kg body weight daily during the entire experimental period. After 31 days the first calf showed fully developed symptoms; two other calves fell ill after another four and eight days respectively. The fourth calf was killed before the disease became clinically manifest. In all calves cerebrocortical necroses were found on histological examination. The clinical picture is described. It was similar to that in spontaneous C.C.N.In the calves exhibiting fully developed symptoms a moderate rise was observed in the pyruvate kinase and a heavy rise in the creatine Phosphokinase activity. On several occasions the total quantity of thiamine (free + phosphorylated) in the blood exceeded the upper normal limit without any extra supply of thiamine being given. Only on one occasion was a lowering observed, to a value just below the lower limit of normal variation.     

Clinical and biochemical findings in bovine cerebrocortical necrosis produced by oral administration of amprolium

Purposing to get some hints on cause and early diagnosis for cerebrocortical necrosis (CCN), CCN was produced in three healthy calves by the oral administration of amprolium. All three calves showed central nervous signs characterized by ataxic gait, clonic spasm, astasia and opisthotonus, from 24 to 49 days after the start of daily administration of 321-418 mg/kg amprolium. They showed bradycardia from about 20 days before the appearance of the nervous signs, which was supposed to be a finding of primary change and to be useful for early diagnosis of CCN. At necropsy of the two calves, large necrotic lesion was found in the cerebral cortex, and tissue thiamine levels decreased significantly, especially in cerebrum and cerebellum. In the other calf, injection with 25 mg thiamine tetrahydrofurfuryl disulfide (TTFD) was proved to be effective for the recovery of clinical signs. No significant changes in thiamine level were recorded in the whole blood, but those in erythrocytes decreased slightly at about a week before the appearance of the clinical signs. No significant alteration of thiamine excretion was observed in urine. Those findings suggest that CCN in calves is caused by thiamine deficiency and that the blood thiamine levels cannot be used for diagnosis of CCN.     

柔嫩艾美耳球虫对盐酸氟乙基硫胺素耐药性的诱导

为评估盐酸氟乙基硫胺素产生抗药性的倾向,采用药物浓度递增法,在实验条件下进行柔嫩艾美耳球虫耐药性的诱导研究。每组动物用10只快大黄肉鸡,每只鸡接种1×105个孢子化卵囊,以50 mg/kg盐酸氨丙啉和盐酸氟乙基硫胺素为起始诱导浓度连续传代,以病变记分、存活率和卵囊数三项指标综合判定耐药性。经过10次传代,盐酸氨丙啉对敏感虫株的ACI为192.6,而对耐药虫株的ACI为109.1;经过12次传代,盐酸氟乙基硫胺素对敏感虫株的ACI为197.5,而对耐药虫株的ACI为110.6,对盐酸氨丙啉耐药虫株的ACI为163.2;结果表明成功地诱导出对盐酸氨丙啉、盐酸氟乙基硫胺素完全耐药的虫株。盐酸氟乙基硫胺素产生耐药速度慢于盐酸氨丙啉,与盐酸氨丙啉有部分交叉耐药性。     

BIOCHEMISTRY OF NATURAL AND AMPROLIUM-INDUCED POLIOENCEPHALOMALACIA IN SHEEP

SUMMARY Polioencephalomalacia (PEM) induced in sheep was compared with the disease found in naturally occurring cases. Blood biochemical indicators measured were pyruvate, lactate, glucose, erythrocyte transketolase (TK) and stimulation of TK by addition of thiamine pyrophosphate (TPP effect). Faeces and rumen contents were assayed for thiaminase activity. The effect of treating affected sheep with thiamine was also noted. It was found that amprolium treatment could induce thrombocytopenia, but once the sheep became accustomed to amprolium in the diet they seemed to be resistant to this effect. In sheep receiving amprolium significant weight losses preceded the onset of clinical signs. Further weight loss continued throughout the recovery period despite removal of amprolium from the diet and treatment with thiamine. Blood glucose was variable, and was elevated only when marked clinical signs were present. Pyruvate and lactate levels showed marked variation throughout the trial. TK values were depressed and TPP effects increased well before the onset of clinical signs, although some naturally occurring cases had normal levels. Faecal thiaminase activity was negligible in all the sheep on the amprolium trial but most field cases had a high level. High faecal thiaminase was observed in about 5% of clinically normal animals from affected flocks. Depression of erythrocyte TK activity coupled with the presence of faecal thiaminase appeared to be the most reliable diagnostic biochemical parameters for PEM. Treatment of PEM affected sheep with thiamine rapidly brought the biochemical status of the animals to normal. However where advanced brain lesions were present the damage was permanent and such sheep treated with thiamine remained partially decorticate.     

Thiamine and its derivates in cattle blood measured by HPLC in healthy animals, in patients suffering from CCN and in their cohorts

The aim of this project was to develop a routine method to measure thiamine and its derivates in blood, to detect the need of vitamin B1-supply in cattle faster and more precise than until now using the transketolase test. For that, thiamine and its derivates (thiaminediphosphate = TDP,-monophosphate = TMP) were analysed in cattle blood by HPLC technique with post-column derivatization. Moreover, the levels in healthy cattle were compared with those in herds with single CCN cases. EDTA-blood-stored up to 10 days by 6 or 20 degrees C- was the most appropriate substrate. Sensitivitiy and specifity were significantly better than (TT, sum of all fractions): 1.95% and 2.10%; thiamine and TMP: 7.02% and 9.17%. The lowest concentration, which could be measured, was 0.5 ng/ml. 72 samples could be analysed daily. In the blood of 280 healthy calves and cows (group A), 201 clinically unsuspicious animals out of stocks with single cases of CCN (group B) and 12 patients with clinical obvious CCN (group C), thiamine and its phosphates were measured and the means calculated (ng/ml). After this, the thiamine status is characterized by (ng/ml): (A) thiamine 13.5 (4.61-28.8),TDP 51.0 (33.1-82.2), TMP 8.79 (2.23-18.1),TT 73.3 (44.6-114); (B) thiamine 8.73 (0.00-20.0), TDP 43.5 (15.5-75.7),TMP 6.15 (0.00-16.6), TT 59.4 (18.7-96.5); (C) thiamine 3.85 (0.00-1 1.5), TDP 14.5 (0.00-28.3), TMP 1.27 (0.00-4.87), TTP 19.6 (4.92-35.9). So, HPLC with post-column derivatization offers a precise, automatically working method for a fast diagnose of thiamine deficiency. The lowest critical value of TT is 40 ng/ml for healthy animals in the blood. The sample (EDTA-blood) can be posted simply at temperatures up to 20 degrees C. In comparison to other measuring methods, this HPLC-technique enables an early recognition of the risk to come down with CCN in endangered cattle herds.     

Thiamine deficiency affects glucose transport and β‐oxidation in rats

Thiamine is recognized as a cofactor for many enzymes involved in intermediary metabolism responsible for energy production. Animal model of thiamine deficiency (TD) included direct evaluation of glucose uptake by estimation of ³H‐deoxyglucose transport across red blood cells membranes and β‐oxidation of fatty acids in isolated leucocytes. Feeding of animals with the thiamine‐deficient diet (0.018 mg/kg diet) for 30 days resulted in disturbances in energy production. The thiamine intake was limited not only by vitamin B₁ deficiency in the diet, but also by time‐dependent drop of feed consumption by rats fed this diet. At the end of experiment, diet consumption in this group of rats was 52% lower than in the control group. This was accompanied by low glucose uptake by erythrocytes of rats suffering vitamin B₁ deficiency for longer time. At the end of experimental period, glucose uptake was over 2 times lower in TD erythrocytes than in control RBC. Such drop of energy production was not compensated by delivery of energy from fatty acid degradation. In leucocytes from TD rats, the β‐oxidation was also suppressed. Observed significant decrease of serum insulin from 2.25 ± 0.25 ng/ml (day 0) to 1.94 ± 0.17 ng/ml (day 30) might have significant impact on observed energy production disorders. The results from this study indicate that the thiamine deficiency significantly reduces feed intake and causes modest abnormalities in glucose and fatty acid utilization.     

硫胺素在鸡肌肉中的沉积规律研究

为研究鸡肌肉组织中硫胺素的沉积规律,以如皋鸡和文昌鸡为素材,分不同家系、周龄、组织进行取样,采用高效液相色谱法检测样品中的硫胺素含量。结果显示,不同品种、家系、周龄及组织间硫胺素含量均存在显著或极显著差异（P<0.05或P<0.01）。此外,硫胺素含量的遗传力为0.494,与体重的遗传相关为-0.404。以上结果初步表明,硫胺素在机体的沉积受遗传因素影响,且表现为一定的时间和组织变化规律,可以通过选择进行提高。     

Clinical signs,MRI features,and outcomes of two cats with thiamine deficiency secondary to diet change

Two cats were presented with vestibular signs and seizures. Both cats were diagnosed with thiamine deficiency. The transverse and dorsal T2-weighted magnetic resonance (MR) images revealed the presence of bilateral hyperintense lesions at specific nuclei of the midbrain, cerebellum, and brainstem. After thiamine supplementation, the clinical signs gradually improved. Repeated MR images taken 3 weeks after thiamine supplementation had started showed that the lesions were nearly resolved. This case report describes the clinical and MR findings associated with thiamine deficiency in two cats.     

Chemotherapeutic efficacy of sulphadimidine, amprolium, halofuginone and chloroquine phosphate in experimental Eimeria bareillyi coccidiosis of buffaloes

Sulphadimidine, amprolium, halofuginone and chloroquine phosphate were administered to buffalo calves 10 days after experimental infection with Eimeria bareillyi. Animals given sulphadimidine or amprolium remained clinically normal and shed only a few oocysts in their faeces. Halofuginone was found partially effective and chloroquine phosphate completely ineffective in preventing faecal oocyst discharge and intestinal lesions. Sulphadimidine and amprolium treated calves gained weight, but chloroquine treated calves suffered progressive weight loss similar to that of infected untreated controls. No significant alterations of haematological values were observed either in the treated calves or in the untreated controls.     

The effect of amprolium and of payzone on the functional state of the thyroid in growing chickens

Amprolium fed to growing chickens in a concentration of 125 p.p.m. in a commercial diet was accompanied by a small but significant decrease in thyroid activity compared with the results for untreated chickens, judged by serum PBI values. The epithelium : colloid relation was lowered. This was presumed to be a consequence of the antagonistic effect of amprolium on thiamine. Payzone feeding, 10 p.p.m. in commercial feed, caused a more pronounced retardation of thyroid activity, judged by the same criteria. A possible connection between the depressed thyroid function and the well-established improvement in weight gain and feed conversion in connection with payzone feeding is discussed.     

Effects of Enterococcus faecium M 74 strain on selected blood and production parameters of laying hens

1. The aim was to evaluate the functional efficiency of a probiotic strain Enterococcus faecium M 74 in the feed on selected biochemical, haematological and production parameters of ISA Brown hens. 2. Feed in the experimental group was enriched with a probiotic preparation containing of 5 × 10(9) viable E. faecium M 74 per g. Blood samples were collected during the egg-laying period at 5 (w5), 25 (w25) and 45 (w45) weeks of production. Body weight, rate of lay and egg weight were recorded every 4 weeks during the 48-week laying period. 3. Significantly lower concentrations of total cholesterol and total lipids in blood plasma were observed in the experimental group at all sampling times compared with their respective controls. Concentrations of triglycerides did not differ. Significantly lower concentrations of plasma calcium were found in the experimental group at w5 and w45. Concentrations of inorganic phosphorus in the experimental group were significantly higher at w25, but significantly lower at w45. Erythrocyte count was significantly higher in the experimental group at w25 and w45 when compared with controls. Leucocyte counts were significantly lower in the experimental group at all sampling times. Significantly lower values of haematocrit at w5 and w45 were observed in the experimental group than in controls. Body weight, the number of eggs and average egg weight were not significantly affected by probiotic addition. 4. In conclusion, the addition of probiotic strain E. faecium M 74 to the feed of ISA Brown hens reduced cholesterol, lipids, calcium, leucocyte counts and haematocrit values in blood plasma in at least two sampling times, while erythrocyte counts were increased. No significant effects of probiotic on triglyceride concentration and egg production parameters were observed.     

Diagnostic aspects of cerebrocortical necrosis.

Specimens from cattle and sheep suspected of having cerebrocortical necrosis (CCN) were studied. Rumenal contents were examined for thiaminase-producing bacteria. Thiaminase activity was assessed in rumenal contents. The thiamine concentration of liver, brain and heart was determined and erythrocyte transketolase assessed. Diagnosis in each case, whether positive or negative for CCN, was decided by histopathological examination. There was a substantial agreement between the biochemical findings and the histological diagnosis indicating that a provisional diagnosis may be made on clinical and biochemical data alone. The findings are discussed in relation to other diseases which have the same neuropathological features. Attempts to isolate thiaminase-producing bacteria, which may be implicated in the aetiology of CCN, were inconclusive.     

Influences on fermentation and thiamine metabolism in bovine rumen fluid (in vitro)--part 3: Effects of artificially contaminated hay with Cladosporium herbarum and Fusarium graminearum, respectively

The influence of contaminated hay [Cladosporium herbarum (CL) and Fusarium graminearum (FU), respectively] on fermentation and thiamine metabolism of bovine rumen content was investigated using the longterm rumen simulation technique (RUSITEC). Six investigation periods 25 days long each were carried out. A nine days feeding period with normal hay was followed by the testphase I (five days) with a mixture of normal and mouldy hay and testphase II (five days) with additive an 0.3 mg thiamine per reaction vessel. The last four days served as regeneration time with normal hay. The following marginal effects of mouldy hay on rumen fermentation patterns could be noted. A) During testphase I: cellulase activity: +10.0% (FU); alternation of the thiamine derivate pattern, but no effect on total thiamine content (CL, FU). B) During testphase II the results were more obvious: bacterial protein synthesis: -22.6% (CL), -24.4% (FU). Alternation of the fatty acid pattern: propionate (-7.30% FU), n-butyrate (-3.90% CL, +3.49% FU), n-valerate (-8.5% CH, FU). Cellulase activity: -17.0% (CL, FU). But no effect on total thiamine (CL, FU); alternation of the thiamine derivate pattern: more non phosphorylate thiamine. The noted effects on rumen fermentation and thiamine metabolism were not severe enough to be responsible for the development of a CCN.     

Relationship between thiamine concentration and fermentation patterns in the rumen fluid of dairy cows fed with graded concentrate levels

The objective of this study was to investigate the relationship between the thiamine concentration and the fermentation patterns [pH, concentration of short-chain fatty acids (SCFA)] in the free liquid (FRL) and particle-associated liquid (PARL) of the rumen in dairy cows fed with graded concentrate levels in the diet. Four ruminally cannulated Holstein cows in mid lactation were fed [semi-ad libitum, 18 kg dry matter (DM)/day] diets consisting of hay (H) and slowly degradable concentrate (C), offered in five different H:C ratios (% DM basis) in the following sequence: period 1, 30:70; period 2, 40:60; period 3, 50:50; period 4, 60:40 and period 5, 75:25. A negative quadratic relationship was observed between thiamine concentration in FRL and intake of dietary thiamine (R(2) = 0.36), of concentrate (R(2) = 0.38) and of digestible non-fibre carbohydrates (R(2) = 0.37). The thiamine concentration in PARL was higher (p < or = 0.05) compared to FRL. The ruminal thiamine concentration correlated negatively to pH and positively to concentrations of SCFA and propionate in the rumen. R(2) of these relationships varied markedly (0.002 and 0.77), depending on time after feeding and fluid digesta compartment. The higher R(2) were observed at 8 and 11 h after the morning feeding for FRL (R(2) 0.46-0.71) and PARL (R(2) 0.41-0.77), respectively.     

Blood thiamine values in captive adult African lions (Panthera leo).

Heparinized whole-blood samples from 22 adult African lions (Panthera leo) fed diets considered nutritionally adequate in 10 American Zoo and Aquarium Association member zoos in North America were provided for this study. Blood thiamine values were estimated using a standard microbiological assay method. The mean +/- standard deviation for blood thiamine values was 249.3 +/- 43.5 nmol/L with a range in values from 160 to 350 nmol/L after exclusion of one outlier. There were no differences (P > 0.05) in the mean blood thiamine values of male and female lions, or of lions that were over and under 10 yr of age. This range (160 to 350 nmol/L) is proposed as a reasonable estimate of the expected range in blood thiamine values for captive adult African lions as currently fed in North American zoos.     

Electroencephalographic observation on sheep and cattle with experimental cerebrocortical necrosis

Nine Suffolk sheep and 4 Holstein cattle were employed to observe electroencephalographic changes in experimental cerebrocortical necrosis. Amprolium (600 mg/kg/day) alone was given intraruminally 6 sheep and 4 cattle, all of which showed neurological signs and abnormal electroencephalograms. Both amprolium and thiamine (200 mg/day) and thiamine (50 mg/day) alone were given 2 and 1 sheep, respectively, as controls, which did not show any abnormal signs. Abnormal electroencephalograms included continuous slow waves and long-lasting spindles, both of which appeared diffusely and were seen subclinically or with neurological signs. The spindles consisted of slow waves or sharp- (or spike-) and-slow-wave complexes with or without convulsive seizures even during the convulsion stage. Conversely, the seizures occurred with the spindles or slow waves.     

Efficacy of amprolium for the treatment of pathogenic Eimeria species in Boer goat kids

This study evaluated the efficacy of two different doses of amprolium in goats heavily infected with pathogenic Eimeria species. Forty Boer goat kids ranging from 3 to 5 months of age with naturally occurring coccidiosis were randomly divided into 2 groups and treated orally with amprolium at doses of 10mg/kg daily for 5 days (n=20) or 50mg/kg daily for 5 days (n=20). The Eimeria oocyst per gram concentrations were significantly reduced on day 7 in the kids that received amprolium at 50mg/kg, however oocyst concentrations were not significantly reduced in goats that received the 10mg/kg dose. Out of 100 Eimeria oocysts identified from a pooled fecal sample, E. christenseni was the most frequently identified (52%) coccidial species present. The results of this trial indicate that amprolium can be an effective treatment for pathogenic Eimeria species in goat kids, however higher and extralabel doses (50mg/kg) should be used.     

Ultrastructural study of experimental cerebrocortical necrosis in calves (author's transl)

Cerebrocortical necrosis (CCN) was experimentally induced in three calves with the thiamine antagonist Amprolium. The calves were followed clinically. At the time of development of typical clinical signs of CCN the calves were killed and necropsied and a complete histopathologic examination was performed. The light microscopic lesions of the cerebral cortex were those of middle laminar necrosis and deep laminar edema. The necrotic zone consisted of fine vesicles and differed only slightly from the edema zone. In the former some neurons were morphologically normal, others showed signs of injury, while in the latter all neurons were injured. The lesions are consistent with those found in natural cases of CCN. Biopsies from the cerebral cortex were taken at the time of early clinical signs. Ultrastructural studies of these biopsies showed no clear difference between the zones of necrosis and edema observed light microscopically. The ultrastructural changes were characterized by dilatations in the neuropil. The cytoplasm and foot plates of the astrocytes had a "watery" appearance and contained dilated mitochondria and large vesicles. The vesicles were continuous with the granular endoplasmic reticulum. It was concluded that the initial morphologic changes in CCN in calves were indicative of a cytotoxic edema of the astrocytes. This may be due to a cytotoxic edema with subsequent loss of cell volume control. The primary astrocytic changes could then lead to neuronal injury.     

Effect of a prebiotic on performance of partridge

An experiment was conducted to investigate the effect of a prebiotic on performance of partridge. The experiment was carried out with a total of eighty‐day‐old male Chukar partridge (Alectoris chukar chukar) chicks in a completely randomized design. The dietary treatments consisted of a control and an experimental treatment, and each treatment was replicated four times with 10 chicks per replicate. The experimental period lasted 16 weeks with a starter period (0–8 weeks) and a grower period (9–16 weeks). The control group was fed the basal diet, while the experimental group was fed the basal diet supplemented with 0.18% prebiotic Aspergillus meal in each period. Performance parameters included body weight gain, cumulative feed consumption and cumulative FCR and were recorded at biweekly intervals. Carcass characteristics and blood parameters were measured at the end of the experiment (week 16). Although the results showed that prebiotic had no significant effect on body weight gain and cumulative feed conversion ratio; however, the supplementation of Aspergillus meal significantly (p < 0.05) decreased cumulative feed consumption. Also prebiotic significantly increased percentages of breast and gastrointestinal tract, decreased percentage of back‐neck, decreased blood triglyceride and total cholesterol content and increased blood calcium content. From this study, it was concluded that dietary supplementation of 0.18% Aspergillus meal might offer beneficial effects on partridge feed consumption, carcass quality and blood cholesterol.     

饲养方式和饲喂水平对鸡肉硫胺素含量及肉质的影响

本文研究了不同饲养方式和饲喂水平对“广西黄”黄羽肉鸡肉质的影响，结果表明，放养(饲喂水平为对照组的50％)肉鸡的肌纤维直径显著增大，密度减小，肌肉脂肪含量明显升高。肌肉肌苷酸、硫胺素含量亦显著升高。