Bovine citrullinaemia: a clinical, pathological, biochemical and genetic study

Calves affected with citrullinaemia are clinically normal immediately after birth. In the majority of calves the clinical course of the disease was similar. Within 24 hours of birth they become depressed; then within 3 to 4 days were observed to wander aimlessly or stand with their head pressed against a wall or fence. By day 4 to 5 they become recumbent, developed convulsions, followed by collapse and death. Oedema of the cerebral cortex is a consistent histological lesion. Citrulline concentration in blood, cerebrospinal fluid, eye fluid and cerebral tissue is greatly elevated. Information gathered from pedigrees of affected calves indicate that the defect is widely disseminated throughout the Australian Friesian population.     

铜缺乏奶牛神经递质酶的组化特征

为探讨铜缺乏对奶牛大脑、小脑及脊髓乙酰胆碱酯酶（ACHEase)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，剖杀后采集大脑、小脑、脊髓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，发病奶牛大脑、小脑及脊髓中ACHEase计数值均显著低于对照组奶牛（P＜0．05）；除大脑外，铜缺额奶牛脊髓上小脑中ACHEase的扫描值也显著低于对照组（P＜0．05）；发病奶牛脊髓小脑中ACHEase的扫描值也低于对照组（P＜0．05）；发病奶牛组织中ACHEase酶颗粒的分布也发生特征性的变化。结论：神经递质酶活性的改变是导致奶牛铜缺乏症病理过程的重要原因。     

Mannosidosis in Galloway calves

Mannosidosis was diagnosed in four stillborn Galloway calves and an autolyzed full-term fetus from experimental matings of carrier animals. Gross lesions were moderate internal hydrocephalus, and pallor and enlargement of the liver and kidneys and arthrogryposis. Histologic changes in the central nervous system of each calf were marked foamy vacuolation of the cytoplasm of neurones in the cerebral cortex, thalamus and brainstem, and vacuolation of the Purkinje cells of the cerebellum. Spheroids were common throughout the brain and there was also consistent severe foamy cytoplasmic vacuolation of renal tubular epithelial cells and hepatocytes. The activities of alpha-mannosidase, the lysosomal enzyme whose activity is deficient in mannosidosis, and activities of five other lysosomal enzymes were compared in brain, liver, and kidney tissues of three mannosidosis-affected calves and normal calf tissues. Tissues from the affected calves had a marked deficiency of alpha-mannosidase activity compared with the normal tissues; the greatest deficiency was in the liver (99%) and brain (98%). Activities of the other lysosomal enzymes were elevated in the affected tissues compared with normal. Mannosidosis is a lysosomal storage disease that results from a defect in glycoprotein metabolism and affects man, Angus and Angus-related breeds of cattle, such as Murray greys, and the cat. The congenital disease is caused by an inherited deficiency of the lysosomal enzyme alpha-mannosidase, and is inherited in an autosomal recessive manner. Mannosidosis was recently reported in a number of aborted and stillborn Australian Galloway calves from an experimental breeding trial. This is more detailed account of the histological and biochemical results obtained during the trial.     

Differences in blood supply to the cerebral cortex between sheep and calves during slaughter

Using methylene blue boli, injected via an intracardiac catheter, the blood supply to the brain in two one- to 10-day-old calves and three adult sheep was studied during and after severance of the common carotid arteries and jugular veins. Passage of dye through cerebral vessels could not be observed in the exposed cerebrum of sheep after bilateral severance of major blood vessels. When vessels were severed on one side only, the passage of dye was noted for at least 53 seconds. In calves, after bilateral severance, sequential boli of dye could be detected passing through the cerebral vessels for more than 100 seconds. These results provide an explanation and support for the belief that there are major differences in the onset of insensibility between sheep and calves subsequent to severance of the common carotid arteries and jugular veins. The differences in blood supply to the brain which were demonstrated could be accounted for by differences in the blood supply to the brain by the vertebral artery in sheep and cattle. The effects of slaughter on the blood supply to the eye are discussed as a possible explanation of the disparity between results of studies in this general field which have used retinal responses in their investigations and those which have not.     

Diprosopia/dicephalia in calves in northern Italy: clinical and aetio-pathological features

Cephalic parapagia, a rare congenital anomaly caused by the fusion of two monozygotic embryos, is characterized by a single body and a spectrum of duplication of craniofacial structures. The authors describe the clinical and pathological aspects of the parapagus conjoined twin defect in nine calves referred to the Department of Animal Pathology, Turin, between 1999 and 2009. The majority of the calves (eight cases) presented two snouts that shared three or four eyes (diprosopia); one calf presented two separate skulls fused at the foramen magnum (dicephalia). Bilateral inferior brachygnathia was observed in four calves. Post-mortem examination of the skull revealed complete brain duplication with fusion at the caudal portion of the brainstem in all calves. Histological features of the cerebral hemispheres and brainstem were normal; moderate disorganization of the cerebellar cortex was noted in two cases. Cardiac malformations were observed in three calves. No aetiologic cause was determined. This article underscores the importance of diprosopia in cattle species and suggests the need for more detailed investigations to better understand its pathogenesis.     

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.     

Acetylcholinesterase activity and lipid peroxidation in the brain and spinal cord of rats infected with Trypanosoma evansi

Neurological and locomotor clinical signs are described in animals infected with Trypanosoma evansi. These disturbances may be related to changes in the amount of acetylcholine (neurotransmitter) in the synaptic cleft. Therefore, changes in acetylcholinesterase (AChE) activity and lipid peroxidation in brain and spinal cord of T. evansi-infected rats were investigated. Each rat was intraperitoneally infected with 10(6) trypomastigotes kept in fresh (group A; n=13) and cryopreserved blood (group B; n=13). Thirteen served as uninfected (not-infected; group C). In days 4 and 30 post-infection (PI) the rats were anesthetized and subsequently decapitated to obtain the brain and the spinal cord (between vertebrae L1 and S2). The brain was removed and dissected (cerebellum, cerebral cortex, striatum and hippocampus) to measure the activity of AChE and lipid peroxidation, determined by TBARS levels. To verify if T. evansi was present in the central nervous system (CNS), brain structures of three rats of each group were processed by PCR T. evansi-specific. AChE activity was significantly increased in all brain structures and decrease in spinal cord in infected rats in 4 PI (P<0.05). The levels of TBARS were decreased in the brain structures, differently from spinal cord, which showed increased lipid peroxidation in 4 PI. The AChE activity in striatum, cerebral cortex, hippocampus and spinal cord reduced concomitantly with the increase of the enzyme in cerebellum of the infected rats (P<0.05), and the TBARS levels increased in cerebellum, striatum and spinal cord of infected rats compared to non-infected animals in 30 PI. The PCR was positive for T. evansi in all structures of the brain, confirming the presence of the parasite in the CNS. Based on the results, we conclude that the changes in AChE activity and lipid peroxidation in the CNS are induced by infection with T. evansi, suggesting that the parasite interferes with the cholinergic neurotransmission in this experimental condition.     

An immunohistochemical and ultrastructural study on age-related astrocytic gliosis in the central nervous system of dogs.

The pattern of astrocytic gliosis (AG) was examined in 2-month-old to 18-year-old dogs using glial fibrillary acidic protein (GFAP) immunohistochemistry and electron microscopy. Coronal sections from various levels of the central nervous system (CNS) were stained with hematoxylin & eosin, Luxol Fast Blue, Nissl, and Bodian in addition to GFAP. A consistent pattern of age-related AG was observed in the dogs. The white matter, cortico-medullary junction, and subcortical nuclei in the cerebrum, central nuclei in the cerebellum, various nuclei in the brain stem, and grey matter of the spinal cord showed even and intense GFAP staining. AG was also prominent in the cerebral and cerebellar cortices and thalamus. Moderate AG was observed in the hippocampus and white matter of the cerebellum and spinal cord. Electron microscopy demonstrated increased number of profiles of degenerative neural components in the vicinity of hypertrophic astrocytes in the cerebral cortex of the aged dogs. Moderate to severe AG was consistently shown in the CNS of the aged dogs. In contrast, young normal dogs showed minimum amounts of GFAP-positive astrocytes in the CNS. These findings suggest that the observed AG in the CNS of the dogs is a morphological expression of aging.     

The histopathogenesis of paralytic rabies in six-week-old C57BL/6J mice following inoculation of the CVS-11 strain into the right triceps surae muscle

A fatal encephalomyelitis was developed after intracerebral and hind limb inoculation of in 6-week-old C57BL/6J mice by the inoculation of fixed rabies virus (CVS-11 strain), intracerebrally and into hind. After the intracerebral inoculation, virus antigens were detected in the cerebral cortex and hippocampus at 2 days postinoculation (PI), and later spread centrifugally to thalamus, brain stem, cerebellum, spinal cord and spinal ganglia. At 4 days PI, severe apoptosis and DNA fragmentation were observed in the hippocampus and cerebral cortex. All mice infected intracerebrally were dead without limb paralysis at from 10 to 11 days PI. In contrast, mice infected with virus intramuscularly were persistently observed virus antigens in the myocytes at the site of inoculation from 2 days PI. At 4 days PI, the antigens were demonstrated in the spinal dorsal root ganglia, spinal cord and muscle spindles without their detection in the cerebrum and hippocampus. There were no apoptosis in the spinal cord and dorsal root ganglia, however hind limb paralysis was found in all infected mice. Hind limb paralysis was progressed to quadriparalysis, and mice were dead from 11 to 13 days PI. From 4 days PI, necrosis of neuron was observed in the the spinal and dorsal ganglia with infiltration of lymphocyte. This study suggested that the necrosis of spinal neurons was more important to cause the paralysis of hind limb rather than the severe cerebral infection and apoptosis in C57BL/6J mice infected with CVS-11 strain. The virus primarily replicated in the muscles was ascended the spinal cord via afferent fibers and retrogradely invaded the cerebrum, and with subsequent spread to muscle spindles.     

Morphologic and immunoperoxidase study of neurologic lesions in naturally acquired rabies of raccoons.

Histopathologic (hematoxylin and eosin [HE]) and immunoperoxidase (streptavidin-biotin complex) methods were used for examination of formalin-fixed tissues of rabid raccoons from an enzootic area of Pennsylvania. Extensive morphologic lesions of rabies encephalitis were present in the cerebrum and the brain stem regions. Negri bodies were detected by both methods and were present in the brain (cerebral cortex, hippocampus, brain stem, cerebellum, and cervical spinal cord) and in the ganglia of the trigeminal nerves. The viral inclusions were also seen in ganglion cells in the tongue, parotid salivary glands, pancreas, intestines, and adrenal glands. These sites were not associated with any inflammatory cellular infiltrate. The immunoperoxidase method was superior to HE for the detection of Negri bodies. Because lesions of rabies encephalitis were consistently observed in the cerebrum, brain stem, and cervical spinal cord regions, these areas of the brain should be included when raccoons are examined by the fluorescent antibody test for rabies.     

Natural scrapie: detection of fibrils in extracts from the central nervous system of sheep

Extracts from the cervical spinal cord and from the medulla, thalamus, cerebellum and cerebral cortex of the brains of 10 sheep, histopathologically confirmed as cases of scrapie, were examined by electron microscopy for the presence of scrapie-associated fibrils. Characteristic fibrils were observed in all the extracts except for that from the thalamus of one sheep. No fibrils were found in any extracts from three control sheep. A comparison of these results with a similar study of 22 cases of bovine spongiform encephalopathy (BSE) suggests that in cases of scrapie the area of the brain chosen for the detection of fibrils is less critical than in cases of BSE, in which fibrils are more readily extracted from areas of the brain stem.     

替来他明诱导大鼠中枢JUN核蛋白表达的研究

通过检测替来他明诱导大鼠中枢神经系统JUN核蛋白的表达,了解替来他明在中枢神经系统的作用部位,探讨替来他明对中枢神经系统的作用机制。72只SD大鼠,随机分为生理盐水组、给药后10、30、60、90、100、120、180min组,每组9只。腹腔注射替来他明50mg／kg后,分别于给药前（生理盐水组）和给药后10、30、60、90、100、120、180min经4％多聚甲醛（0．1mol／L,PB,pH7．4）灌注后,取脑,将脑分为大脑皮层、海马、丘脑、小脑、脑干等5个脑区置于20％蔗糖溶液中24h（4℃）脱水。冰冻切片,片厚10μm。免疫组织化学法（Elivision法）检测JUN核蛋白,统计各组大鼠脑片的JUN免疫反应阳性神经元的分布。对照组中仅发现少量JUN核蛋白,试验组中JUN核蛋白在大脑皮层、海马、丘脑、小脑、脑干内都有表达。替来他明腹腔注射10min后JUN核蛋白表达开始增加,60min表达至高峰,90min表达下降,180min下降至基线水平,与给药前相比无显著性差异（P〉0．05）。替来他明能诱导大鼠大脑皮层、海马、丘脑、小脑、脑干c-iun基因的表达,大脑皮层、海马、丘脑、小脑和脑干是替来他明的作用位点。     

Congenital tremor in Holstein Friesian-cattle.

In a female family line of Holstein-Friesian cattle a series of trembling calves was born within a period of ten years. All trembling animals were male; female calves appeared normal. At necropsy of one calf severe degenerative lesions of spinal cord and brain white matter were observed. Both myelin and axons were lost and some macrophages occurred within digestion chambers. Obviously degenerated neurones were not found. The lesion was interpreted as a single recessive sex-linked hereditary trait.     

Akabane virus infection in the pregnant ewe. 2. Pathology of the foetus

A strain of Akabane virus (CSIRO 16) isolated from Culicoides brevitarsis was given three different passage treatments in the laboratory and then inoculated into ewes that were 32 to 36 days pregnant. The foetuses from these ewes were examined between the 69th and 106th days of gestation. The 39 infected ewes produced 55 foetuses of which 44 (80%) had severe developmental defects. Arthrogryposis and agenesis of the brain or hydranencephaly, were present in 43 of the foetuses. Other gross defects affecting variable proportions of foetuses were porencephaly, brachygnathism, scoliosis, hypoplasia of the lungs and agenesis or hypoplasia of the spinal cord. Histopathological findings covered a wide spectrum of defects that have previously been considered to occur over an extended range of foetal ages. These defects included skeletal muscle atrophy and degeneration, and in the brain, particularly in the cerebrum, cystic areas and malacia, general oedema, subependymal gliosis, perivascular cuffing and mineralised plaques. Similar lesions were seen in the pons and cerebellum. Extensive lesions, with and without inflammation were seen in the spinal cord.     

Neospora caninum associated with epidemic abortions in dairy cattle: The first clinical neosporosis report in Turkey

Neospora caninum, a protozoan parasite, has been considered as one of the most important etiological agents responsible for abortion in dairy cattle throughout the world since it was first identified in dogs in 1988. In this report, characteristics of neosporosis, detected in a dairy cow ranch having epidemic abortions as high as 18.4%, were described. Blood samples were collected from 25 infertile or aborted dairy cattle, 6 calves born in 2006 and 40 heifers that were born in 2005 and raised in the same ranch. Necropsy was conducted in a 20-day-old Simmental calf that exhibited neurological signs including incoordination, head shaking, hyperextension in forelimbs and hindlimbs and tremor. The seroprevalance in aborted or infertile dairy cattle, heifers, and calves was 60%, 40%, and 33.3%, respectively. The mothers of seropositive two calves including clinically affected calf and its dam were N. caninum seropositive. In immunoperoxidase examinations, N. caninum antigen immunopositivity was observed in the degenerative and necrotic neurons in the brain, cerebellum as well as neurons in dorsal root ganglia of the cervical and thoracic regions of the spinal cord. In the heart, myocytes and Purkinje cells exhibited granular and linear patterns of immunoreactivity. Striated myofibers around the eyeball also showed immunolocalization for N. caninum antigen. Ultrastructurally, tachyzoites with typical apical complex, rhoptries and double-layered parasitic membrane were detected in the brain and heart sections. In conclusion, this report described clinical neosporosis for the first time in Turkey with tissue localization of the causative agents. This scientific communication also discusses the possible impact of cattle neosporosis by clinical, serologic and pathologic evidences collected from the survey of calves born in two successive generations in a ranch.     

Demonstration of Akabane virus antigen using immunohistochemistry in naturally infected newborn calves

Eight newborn calves showing ataxia were necropsied and examined histologically. Six of seven cerebrospinal fluid samples collected from these animals had neutralizing antibody for Akabane virus (AKV). All examined calves had nonsuppurative encephalomyelitis, localized mainly in the midbrain and spinal cord. Corresponding to the encephalitic lesion, AKV antigen was demonstrated in neuroglial cells in the brain stem and neuronal cells in the ventral horn of the spinal cord. This is the first study to demonstrate AKV antigen by immunohistochemistry in naturally infected newborn calves.     

Congenital tremor in Holstein Friesian‐cattle

Summary

In a female family line of Holstein‐Friesian cattle a series of trembling calves was born within a period of ten years. All trembling animals were male; female calves appeared normal At necropsy of one calf severe degenerative lesions of spinal cord and brain white matter were observed Both myelin and axons were lost and some macrophages occurred within digestion chambers. Obviously degenerated neurones were not found The lesion was interpreted as a single recessive sex‐linked hereditary trait.     

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.