Specialised Vascular Structures in the Pododerma of the Camel Footpad (Camelus dromedarius)

Transversal, horizontal and sagittal sections of the brain were stained by the ancient but efficient Mulligan method, a procedure that establishes a clear macroscopic difference between the white and grey substances. Different structures of each section were studied and most of the details were identified and named according to the NAV. All sections were projected onto the whole brain. By means of this easy and basic procedure the students increase their understanding of (1) the size and/or the form and/or the topography of several prominent structures of the brain, (2) the general distribution of the substancia alba and grisea, and they begin to understand the complexity of the brain.     

Normal feline brain: clinical anatomy using magnetic resonance imaging

The purpose of this study was to provide a clinical anatomy atlas of the feline brain using magnetic resonance imaging (MRI). Brains of twelve normal cats were imaged using a 1.5 T magnetic resonance unit and an inversion/recovery sequence (T1). Fourteen relevant MRI sections were chosen in transverse, dorsal, median and sagittal planes. Anatomic structures were identified and labelled using anatomical texts and Nomina Anatomica Veterinaria, sectioned specimen heads, and previously published articles. The MRI sections were stained according to the major embryological and anatomical subdivisions of the brain. The relevant anatomical structures seen on MRI will assist clinicians to better understand MR images and to relate this neuro-anatomy to clinical signs.     

Demonstration of pseudorabies viral antigen in thick and ultrathin tissue sections of young pigs using an immunogold method

Pseudorabies (Aujeszky's) virus antigens were labeled in thick and ultrathin tissue sections of young pig brain and liver tissue, using an indirect immunogold method. Antigens were tagged with 20 nm gold particles. Controls proved the specificity of the reaction in paraffin sections and ultrathin epoxy sections. Immunogold staining was compared with immunoperoxidase staining in paraffin sections. In ultrathin sections stained with the immunogold method, the gold particles were present on viral nucleocapsids and viral envelopes, as well as on a number of other intracellular structures. These included the inner nuclear membrane, the nucleoplasm, intranuclear filaments, the endoplasmic reticulum, and free cytoplasmic polyribosomes. Gold particles were absent on mitochondria and microtubules. In paraffin sections, immunogold labeling for pseudorabies virus antigen was less sensitive than immunoperoxidase staining. Immunogold staining of ultrathin tissue sections can yield additional information on virus-host cell interactions.     

Distribution of protease-resistant prion protein and spongiform encephalopathy in free-ranging mule deer (Odocoileus hemionus) with chronic wasting disease

Serial sections of brain and palatine tonsil were examined by immunohistochemical staining (IHC) using monoclonal antibody F89/160.1.5 for detecting protease-resistant prion protein (PrP(res)) in 35 hunter-killed mule deer (Odocoileus hemionus) with chronic wasting disease. Serial sections of brain were stained with hematoxylin and eosin and examined for spongiform encephalopathy (SE). Clinical signs of disease were not observed in any of these deer. On the basis of the location and abundance of IHC and the location and severity of SE, deer were placed into four categories. Category 1 (n = 8) was characterized by IHC in the palatine tonsil with no evidence of IHC or SE in the brain. Category 2 (n = 13) was characterized by IHC in the palatine tonsil and IHC with or without SE in the dorsal motor nucleus of the vagus nerve (DMNV). Category 3 (n = 2) was characterized by IHC in the palatine tonsil, IHC with SE in the myelencephalon, and IHC without SE in the hypothalamus. Category 4 (n = 12) was characterized by IHC in the palatine tonsil and IHC with SE throughout the brain. Category I may represent early lymphoid tissue localization of PrP(res). The DMNV appears to be the most consistent single neuroanatomic site of detectable PrP(res). Categories 2-4 may represent a progression of spread of PrP(res) and SE throughout the brain. IHC in tonsil and brain and SE in brain were not detected in 208 control deer.     

Immunohistochemical detection of cellular prion protein (PrPc) in the rat central nervous system

A simple conventional method of immunohistochemistry (i.e. fixing the frozen sections in cold methanol) was used to determine the immunolocalization of cellular prion protein (PrPc), with good results. In the rat cerebrum, the cytoplasm of neural cells in the cortex and corpus stratum, pia mater, membrane limitans gliae superficialis, choroid plexus and blood vessel wall were immunostained. The formation of network structures of immunostained neural and/or glial fibers in the cerebral cortex was also observed. These immunostained network structures of neural and/or glial fibers were also observed in cultured neural cells. The results suggest that fixation of frozen sections and cultured cells with cold methanol is a useful method for detecting the immunolocalization of PrPc and that PrPc exists in the various components of the central nervous system of the rat.     

Comparison of histological lesions and immunohistochemical staining of proteinase-resistant prion protein in a naturally occurring spongiform encephalopathy of free-ranging mule deer (Odocoileus hemionus) with those of chronic wasting disease of captive mule deer

In this investigation, the nature and distribution of histologic lesions and immunohistochemical staining (IHC) of a proteinase-resistant prion protein were compared in free-ranging mule deer (Odocoileus hemionus) dying of a naturally occurring spongiform encephalopathy (SE) and captive mule deer dying of chronic wasting disease (CWD). Sixteen free-ranging deer with SE, 12 free-ranging deer without SE, and 10 captive deer with CWD were examined at necropsy. Tissue sections were stained with hematoxylin and eosin, and duplicate sections were stained with a monoclonal antibody (F89/160.1.5). Histological lesions in the free-ranging deer with SE and captive deer with CWD were found throughout the brain and spinal cord but were especially prominent in the myelencephalon, diencephalon, and rhinencephalon. The lesions were characterized by spongiform degeneration of gray matter neuropil, intracytoplasmic vacuolation and degeneration of neurons, and astrocytosis. IHC was found throughout the brain and retina of deer with SE and CWD. Positive IHC was found in lymphoid tissue of deer with SE and CWD. Histologic lesions and IHC were not found in multiple sections of integument, digestive, respiratory, cardiovascular, endocrine, musculoskeletal, and urogenital systems of deer with SE or CWD. Comparison of histologic lesions and IHC in tissues of free-ranging deer with those of captive deer provides strong evidence that these two diseases are indistinguishable morphologically.     

TOMY OF THE CANINE BRAIN USING HIGH RESOLUTION OMPUTED TOMOGRAPHY*

High resolution computed tomography (CT) was used to determine the normal appearance of the brain of an adult Beagle dog. Objects as small as 0.6 mm for bony structures (high contrast) and 1.5–2.0 mm for soft tissue structures (low contrast) could be resolved in the CT images. Multiplanar imaging using direct transverse and reformatted dorsal and sagittal images made it possible to obtain a three dimensional presentation of anatomy. Selective viewing, where CT number window and level settings were varied, was used to optimize visualization of specific brain structures. Normal high contrast components, cerebrospinal fluid, and osseous land-marks, were important aids in identification of various intracranial structures. Quantitative densitometry was performed to characterize various regions of the brain in terms of their x-ray attenuation values or CT numbers. This study indicated that high resolution CT provides a qualitative and quantitative appraisal of the canine brain that is unavailable using conventional radiographic technics.     

Binding of curcumin to senile plaques and cerebral amyloid angiopathy in the aged brain of various animals and to neurofibrillary tangles in Alzheimer's brain

The binding of curcumin to senile plaques (SPs) and cerebral amyloid angiopathy (CAA) was examined in the aged brain of various animal species and a human patient with Alzheimer's disease (AD), together with its binding to neurofibrillary tangles (NFTs). Brain sections were immunostained with anti-amyloid β protein 1-42 (Aβ42) and anti-amyloid β protein 1-40 (Aβ40) antibodies. These sections were also stained with alkaline Congo red, periodic acid-methenamine silver (PAM), and curcumin (0.009% curcumin solution) with or without formic acid pretreatment. The sections from the AD brain were also immunostained for anti-paired helical filament-tau (PHF-tau), and were stained with Gallyas silver for NFTs. Some SPs in the AD, monkey, dog, bear, and amyloid precursor protein transgenic mouse (APP Tg-mouse) brains contained congophilic materials, and were intensely positive for curcumin. In addition, curcumin labeled some diffuse SPs negative for Congo red in the AD, monkey, bear, and APP Tg-mouse brains. In all animals, CAA was intensely positive for both Congo red and curcumin. The specific curcumin staining activity was lost by formic acid pretreatment. In the AD brain, NFTs positive for PHF-tau and Gallyas silver were moderately stained with curcumin. These findings indicate that curcumin specifically binds to the aggregated Aβ molecules in various animals, and further to phosphorylated tau protein, probably according to its conformational nature.     

Morphological characteristics of the forebrain in the donkey (Equus asinus): A compared atlas of magnetic resonance imaging and cross-sectional anatomy

The brain is the most essential part of the central nervous system which regulates and coordinates all body activities. Based on its phylogenetic development from the neural tube, the brain is divided into rhombencephalon (hindbrain), mesencephalon (midbrain) and prosencephalon (forebrain). The present study is achieved to describe the morphological characteristics of the normal forebrain in the donkey using the matched magnetic resonance imaging (MRI) and cross-sectional anatomy. Ten cadaveric heads of healthy adult donkeys of both sexes were used. Two heads were examined using a 1.5 Tesla MRI scanner, and the brains of the other heads were gently extracted; six brains were sectioned into transverse, dorsal and sagittal slices, and two brains were grossly inspected. MR images were selected in correlation to their closely corresponding gross sections. Both cross-sectional anatomy and MRI scans showed extensive gyration of the neocortex. The forebrain structures appeared with variable intensities on three sequences, Flair, T1-weighted and T2-weighted MRI, enabling comprehensive evaluation of the relevant neuroanatomical structures. The present study provided a precise neuroanatomical atlas of the forebrain in the donkey which could help in the quick and efficient interpretation of clinical diseases of the forebrain, localization of the forebrain functions and evolutionary neurobiology.     

Primary canine and feline nervous system tumors: intraoperative diagnosis using the smear technique

The recent application of neuroimaging techniques in veterinary neurology has led to the accurate localization of many types of intracranial lesions but has also created a clinical need, particularly with brain tumors, for a specific intraoperative diagnosis. For human brain tumors, a smear technique has been used successfully for many years to provide an extremely rapid, highly accurate intraoperative diagnosis. In similar smear preparations of intracranial lesions, obtained either by computed tomography (CT)-guided stereobiopsy or from a craniotomy, we have described distinguishing cytologic features of some primary spontaneous nervous system tumors in 80 dogs and 13 cats. A final diagnosis was confirmed by evaluation of paraffin-embedded sections from the same sample and, when appropriate, by immunocytochemical staining. Preliminary findings indicate that, in dogs and cats, this procedure is useful for rapid, accurate intraoperative diagnosis of many primary nervous system tumors. The distinguishing features of the canine and feline tumors bear a remarkably close resemblance to their human counterparts.     

Ventral surgical approach to the caudal brain stem in dogs

OBJECTIVE: To develop a safe neurosurgical procedure that accessed the ventral pons and medulla of the dog primarily for the removal of brain stem neoplasms. STUDY DESIGN: In vivo study. METHODS: A cadaver study was performed on mesocephalic dog heads to develop a neurosurgical approach to the ventral brain stem. Based on this study, an approach to the ventral brain stem was developed by basioccipital craniectomy. This procedure was performed on 4 young neurologically normal Beagle dogs to assess morbidity and mortality associated with the procedure. Morbidity was evaluated by subjective criteria, daily complete neurologic examinations, comparison of preoperative and postoperative brain stem auditory evoked response (BAER) tests, and postmortem examinations. RESULTS: Three dogs developed a transient cough but were neurologically normal after surgery. One dog was euthanatized within 12 hours of surgery because of severe postoperative morbidity associated with basilar artery disruption due to improper development of the craniectomy. Prolongations of postoperative BAER latencies were observed in 2 dogs but did not appear to be associated with clinical deficits or histopathologic changes in the brain stem. Minimal histopathologic changes were observed except in the dog with basilar artery disruption. Results of this study indicate that, although technically challenging, this procedure can be performed with minimal morbidity. CLINICAL IMPLICATIONS: The main indication for this procedure is surgical reduction or biopsy of ventrally located brain stem neoplasms in dogs. The major disadvantage is anatomic restrictions that prevent access to laterally oriented ventral brain stem masses.     

Dual Sarcocystis neurona and Toxoplasma gondii infection in a Northern sea otter from Washington state, USA

Dual Sarcocystis neurona and Toxoplasma gondii infection was observed in a Northern sea otter from Washington, USA. The animal was found stranded, convulsed, and died shortly thereafter. Encephalitis caused by both S. neurona and T. gondii was demonstrated in histological sections of brain. Immunohistochemical examination of sections with S. neurona specific antisera demonstrated developmental stages that divided by endopolygeny and produced numerous merozoites. PCR of brain tissue from the sea otter using primer pairs JNB33/JNB54 resulted in amplification of a 1100 bp product. This PCR product was cut in to 884 and 216 bp products by Dra I but was not cut by Hinf I indicating that it was S. neurona [J. Parasitol. 85 (1999) 221]. No PCR product was detected in the brain of a sea otter which had no lesions of encephalitis. Examination of brain sections using T. gondii specific antisera demonstrated tachyzoites and tissue cysts of T. gondii. The lesions induced by T. gondii suggested that the sea otter was suffering from reactivated toxoplasmosis. T. gondii was isolated in mice inoculated with brain tissue. A cat that was fed infected mouse brain tissue excreted T. gondii oocysts which were infective for mice. This is apparently the first report of dual S. neurona and T. gondii in a marine mammal.     

Use of an immunoperoxidase test for the detection of bovine herpesvirus-1 in aborted fetal tissue

An indirect immunoperoxidase (IP) procedure using a specific monoclonal antibody and an avidin-biotin-peroxidase complex was developed and applied to detect virus antigen in formalin-fixed, paraffin-embedded tissue sections. This IP procedure was compared with currently used diagnostic tests for detection of virus-induced abortions caused by bovine herpesvirus-1 (BHV-1). The IP procedure was applied to detect BHV-1 antigen in sections of liver and lung from 87 aborted fetuses. Sixteen of these cases were positive for viral antigen by IP staining. Sections from both liver and lung were positive in 15 of the 16 cases. A fluorescent antibody test (FA), which was applied to acetone-fixed frozen sections of liver and lung, gave positive results on 12 of the 87 fetuses, 11 of which were also positive by IP. Seven of the 12 FA-positive cases were positive on both sections of liver and lung. When FA and IP were compared. FA had a sensitivity of 67% and IP had a sensitivity of 94%. Virus was isolated from one of the 67 cases tested. The tissues in which antigen was most frequently detected by IP were liver, lung, and kidney. Distinct multifocal staining was seen in positive sections of all these tissues.     

Isolation of Neospora caninum from the brain of a naturally infected adult dairy cow

Neospora caninum was isolated from the brain of a 2-year-old dairy cow that had aborted confirmed N. caninum-infected fetuses on two occasions. The cow had an indirect fluorescent antibody titer of 1:1600 to N. caninum. The cow was killed 24 days after its second abortion and the brain was bioassayed for N. caninum in nude mice. Multifocal areas of perivascular cuffing and glial nodules were observed in the cerebrum and mesencephalon of the cow, but N. caninum was not identified in histological sections of the brain. All three nude mice inoculated with brain homogenate of the cow, developed emaciation and paralysis. Microscopical examination of the nude mice revealed systemic N. caninum infection with demonstrable tachyzoites in various organs. The parasites isolated from fresh mouse brain were transferred successfully into Vero cell cultures. PCR procedure on the purified tachyzoites obtained from the Vero cell cultures amplified the specific DNA sequence for N. caninum.     

Distribution of lesions in fetal brains following experimental infection of pregnant sheep with Toxoplasma gondii

Six ovine fetal brains were harvested 33 to 35 days postchallenge from 5 ewes, each of which was given 3000 Toxoplasma gondii oocysts on day 90 of pregnancy. Histopathologic examination of transverse sections taken at 13 levels in the fetal brains revealed the presence of toxoplasmosis-related lesions in all 6 brains. However, lesions were not randomly distributed (P = .007); they were most numerous at the level of the optic tract, the rostral margin of the pons, and 4 mm caudal to the ansate sulcus and were absent in all sections at the level of the caudal cerebellum. Lesion distribution may be due to hemodynamic factors, differences in the expression of endothelial surface receptor molecules at the level of the blood-brain barrier, or the presence of localized permissive/inhibitory factors within the brain. The results have implications for the selection of areas of brain from aborted ovine fetuses to be examined histopathologically for laboratory diagnosis.     

犬瘟热的微生物学诊断研究

对临床诊断为犬瘟热的１４例病犬，取其脑组织，运用细胞培养、合胞体检查、包涵体检查、免疫荧光试验、电镜观察等５种检测方法，就犬瘟热的微生物学诊断进行了系统的研究。结果表明，犬脑组织块与猫胚（ＦＥ）细胞或非洲绿猴肾（Ｖｅｒｏ）细胞共同培养，盲传的培养物出现不稳定的细胞病变，通过对不同代次的培养物检查包涵体，并作超薄切片或负染，电镜观察犬瘟热病毒（ＣＤＶ）粒子，证实分离到ＣＤＶ野毒。病犬脑组织切片经ＨＥ染色检查包涵体及用ＣＤＶ荧光抗体直接法检测脑抹片及接种犬脑的细胞培养物，均获得一定的阳性结果。建立的改良离子捕获电镜法，用于检测犬脑匀浆和犬脑接种细胞培养物，与离子捕获电镜法、免疫电镜法及直接电镜法相比，效果更为理想。１４例临床诊断为犬瘟热的病犬，综合运用上述微生物学手段检测，结果为１２例阳性，２例疑似。     

CT-GUIDED BRAIN BIOPSY USING A MODIFIED PELORUS MARK III STEREOTACTIC SYSTEM: EXPERIENCE WITH 50 DOGS

This report describes the results of CT-guided stereotactic brain biopsies performed on 50 consecutive dogs using a modified Pelorus Mark III Stereotactic System. Based on available histopathologic samples (stereotactic biopsy [n = 50], surgery [n = 17], necropsy [n = 9]) the patient population consisted of 34 dogs with primary brain tumors, 2 with invasive nasal adenocarcinomas, and 13 with non-neoplastic brain lesions. Brain tissue was not obtained from one dog. In 22 dogs a final diagnosis was made from tissue subsequently obtained from surgical resection or at necropsy. The final diagnosis was in agreement with the stereotactic biopsy diagnosis in 20 of these 22 dogs. In 17 other dogs without follow-up, stereotactic biopsy provided a diagnosis of a specific primary brain tumor subtype. Postoperative complications associated with the biopsy procedure were assessed in 41 dogs. The other 9 dogs either went directly to surgery (n = 7) or were killed (n = 2) immediately after the biopsy procedure. Thirty-six dogs recovered without apparent clinical complications. Postoperative clinical complications in the remaining 5 dogs included transient epistaxis (1 dog), transient exacerbation of cerebellar signs (1 dog), obtundation progressing to coma (1 dog), and medically uncontrollable seizures (2 dogs). The latter 3 dogs with severe neurologic complications all had large primary brain tumors and had been receiving high doses of phenobarbital and glucocorticoids to control seizures at the time of biopsy. These results suggest that this CT-guided biopsy procedure can provide an accurate pathologic diagnosis of brain lesions detected by CT and MR neuroimaging. Further refinement of both technique and case selection is expected to reduce the rate of clinical complications and to improve the accuracy of the procedure.