Reduced retinal transduction and enhanced transgene-directed immunogenicity with intravitreal delivery of rAAV following posterior vitrectomy in dogs.

Adeno-associated virus (AAV) vector-based gene therapy is a promising treatment strategy for delivery of neurotrophic transgenes to retinal ganglion cells (RGCs) in glaucoma patients. Retinal distribution of transgene expression following intravitreal injection (IVT) of AAV is variable in animal models and the vitreous humor may represent a barrier to initial vector penetration. The primary goal of our study was to investigate the effect of prior core vitrectomy with posterior hyaloid membrane peeling on pattern and efficiency of transduction of a capsid amino acid substituted AAV2 vector, carrying the green fluorescent protein (GFP) reporter transgene following IVT in dogs. When progressive intraocular inflammation developed starting 4 weeks post IVT, the study plan was modified to allow detailed characterization of the etiology as a secondary goal. Unexpectedly, surgical vitrectomy was found to significantly limit transduction, whereas in non-vitrectomized eyes transduction efficiency reached upwards to 37.3% of RGC layer cells. The developing retinitis was characterized by mononuclear cell infiltrates resulting from a delayed-type hypersensitivity reaction, which we suspect was directed at the GFP transgene. Our results, in a canine large animal model, support caution when considering surgical vitrectomy before IVT for retinal gene therapy in patients, as prior vitrectomy appears to significantly reduce transduction efficiency and may predispose the patient to development of vector-induced immune reactions.

Immunohistochemical characterization and morphometric analysis of macrophages in rat mammary tumors.

Macrophages are an important leukocyte component of the microenvironment of neoplasms. Macrophages have classically been subdivided into M1 and M2, depending on their roles in immune response, wound healing, and promotion or inhibition of tumor growth. In human breast cancer, increased presence of M2 macrophages has been associated with poor prognosis. The authors hypothesized that rat mammary carcinomas have increased macrophage influx compared to benign mammary proliferative lesions and normal mammary glands as well. In humans, both M1 and M2 macrophages express CD68, while CD163 is expressed primarily by M2 macrophages. Based on a single immunolabeling protocol with anti-CD68 and anti-CD163 antibodies, the extent of macrophage influx was investigated by morphometry to quantitate the immunopositive cells in normal rat mammary glands, benign mammary proliferative lesions, and mammary carcinomas. In mammary carcinomas, there was significantly higher percentage of CD68+ cells compared to benign mammary proliferative lesions and normal mammary glands. There was also higher percentage of CD163+ cells in mammary carcinomas compared to benign mammary proliferative lesions. Thus, increase in CD68+ and CD163+ macrophages corresponded to increased malignancy of rat mammary tumors in this study.

Megaesophagus in Friesian horses associated with muscular hypertrophy of the caudal esophagus.

Friesian horses have a perceived high rate of congenital or hereditary diseases, including megaesophagus, that may lead to choke and death. A retrospective study was performed to determine the prevalence and pathologic characteristics of esophageal disease in 852 horses, including 17 Friesians, that had been necropsied over a 6-year period at the Diagnostic Center for Population and Animal Health. Forty-two horses had grossly described esophageal lesions (25 muscular hypertrophy, 7 hemorrhage, 6 megaesophagus, 4 erosion/ulceration, 3 obstruction, 2 tears, 2 secondary neoplasms, 2 lymphoid patches, 1 thin wall, 1 esophagitis). Some of these lesions occurred concurrently in the same horse. Ten of these horses died or were euthanatized because of severe esophageal disease (6 megaesophagus causing tears in 2 horses, 3 esophageal obstruction with food bolus, and 1 esophagitis). All 6 horses with megaesophagus were Friesians. No cause for megaesophagus was noted in the necropsy reports; however, 5 of these 6 Friesians had marked caudal esophageal muscular hypertrophy (wall thickness: 1.9 ± 0.3 cm). Microscopic review of the esophagus of these Friesians confirmed smooth muscle hypertrophy, with no obvious fibrosis, degeneration, or loss of myenteric plexi. Unlike the Friesians, the 4 non-Friesian horses with severe esophageal disease had esophageal obstruction with an intraluminal food bolus or severe esophagitis. None had caudal esophageal muscular hypertrophy. It is concluded that in comparison to other horse breeds, Friesians have a higher prevalence of severe esophageal disease, specifically megaesophagus, that is commonly associated with marked caudal muscular hypertrophy.

Beyond H&E: integration of nucleic acid-based analyses into diagnostic pathology.

Veterinary pathology of infectious, particularly viral, and neoplastic diseases has advanced significantly with the advent of newer molecular methodologies that can detect nucleic acid of infectious agents within microscopic lesions, differentiate neoplastic from nonneoplastic cells, or determine the suitability of a targeted therapy by detecting specific mutations in certain cancers. Polymerase chain reaction-based amplification of DNA or RNA and in situ hybridization are currently the most commonly used methods for nucleic acid detection. In contrast, the main methodology used for protein detection within microscopic lesions is immunohistochemistry. Other methods that allow for analysis of nucleic acids within a particular cell type or individual cells, such as laser capture microdissection, are also available in some laboratories. This review gives an overview of the factors that influence the accurate analysis of nucleic acids in formalin-fixed tissues, as well as of different approaches to detect such targets.

Current state of knowledge on porcine circovirus type 2-associated lesions.

Porcine circovirus type 2 (PCV2), a small single-stranded DNA virus, was initially discovered in 1998 and is highly prevalent in the domestic pig population. Disease manifestations associated with PCV2 include postweaning multisystemic wasting disease (PMWS), enteric disease, respiratory disease, porcine dermatitis and nephropathy syndrome (PDNS), and reproductive failure. Although these clinical manifestations involve different organ systems, there is considerable overlap in clinical expression of disease and presence of lesions between pigs and within herds. It is now widely accepted that PCV2 can be further subdivided into different types, of which PCV2a and PCV2b are present worldwide and of greatest importance. This review will focus on PCV2-associated lesions in different organ systems.

Cutaneous mast cell tumor with epitheliotropism in 3 dogs.

Epitheliotropism is an important diagnostic feature of cutaneous epitheliotropic lymphoma and canine cutaneous histiocytoma; however, although noted in certain feline mastocytic diseases, it has not been considered a feature of canine cutaneous mast cell tumor. In this study, 3 canine cutaneous mast cell tumors had epitheliotropic invasion of neoplastic mast cells into the epidermis and follicular epithelium. This unusual histologic finding was characterized by infiltrates of individual and clusters of neoplastic mast cells in the stratum basale and stratum spinosum. The mast cell origin of these cells was documented by demonstration of metachromasia with Giemsa stain and positive immunoreactivity to KIT protein. On the basis of these findings, mast cell tumors should be included in the differential diagnosis for canine cutaneous round cell neoplasms that infiltrate the epidermis.

Somatotroph pituitary tumors in budgerigars (Melopsittacus undulatus).

A series of 11 pituitary tumors in budgerigars were classified on the basis of their clinical, gross, microscopic, and immunohistochemical characteristics. Affected birds were young to middle-aged. Clinically, neurologic signs--including difficulties flying, ataxia, and blindness--were most commonly reported. Additional clinical signs included weight loss, abnormal feathers or molting, increased respiratory efforts, and exophthalmos. Nine birds were diagnosed with chromophobic pituitary adenomas, and 2 birds had chromophobic pituitary carcinomas. Only 1 tumor was delimited to the pituitary gland; the other 10 variably invaded the brain, skull, and retrobulbar space. Distant metastases were identified in 2 birds. All tumors were immunohistochemically strongly positive for growth hormone, consistent with the diagnosis of somatotroph tumors. The common occurrence and early onset may suggest a genetic predisposition of budgerigars to develop somatotroph pituitary tumors with a high incidence of local invasion and with metastatic potential.

Proposal of a 2-tier histologic grading system for canine cutaneous mast cell tumors to more accurately predict biological behavior.

Currently, prognostic and therapeutic determinations for canine cutaneous mast cell tumors (MCTs) are primarily based on histologic grade. However, the use of different grading systems by veterinary pathologists and institutional modifications make the prognostic value of histologic grading highly questionable. To evaluate the consistency of microscopic grading among veterinary pathologists and the prognostic significance of the Patnaik grading system, 95 cutaneous MCTs from 95 dogs were graded in a blinded study by 28 veterinary pathologists from 16 institutions. Concordance among veterinary pathologists was 75% for the diagnosis of grade 3 MCTs and less than 64% for the diagnosis of grade 1 and 2 MCTs. To improve concordance among pathologists and to provide better prognostic significance, a 2-tier histologic grading system was devised. The diagnosis of high-grade MCTs is based on the presence of any one of the following criteria: at least 7 mitotic figures in 10 high-power fields (hpf); at least 3 multinucleated (3 or more nuclei) cells in 10 hpf; at least 3 bizarre nuclei in 10 hpf; karyomegaly (ie, nuclear diameters of at least 10% of neoplastic cells vary by at least two-fold). Fields with the highest mitotic activity or with the highest degree of anisokaryosis were selected to assess the different parameters. According to the novel grading system, high-grade MCTs were significantly associated with shorter time to metastasis or new tumor development, and with shorter survival time. The median survival time was less than 4 months for high-grade MCTs but more than 2 years for low-grade MCTs.

Vascular lesions in pigs experimentally infected with porcine circovirus type 2 serogroup B.

Vasculitis is a hallmark lesion of the severe form of systemic porcine circovirus-associated disease (PCVAD). In 2 experimental studies with porcine circovirus type 2 serogroup b (PCV2b), 2 pigs developed fatal PCVAD with acute vasculitis, and 5 related pigs developed chronic lymphohistiocytic and plasmacytic peri- and endarteritis. Five of these pigs (1 with acute vasculitis and 4 with chronic vasculitis) had also been inoculated with bovine viral diarrhea virus type 1 (BVDV1) or BVDV1-like virus. Vascular lesions were similar, independent of whether pigs had been inoculated singly with PCV2b or dually with PCV2b and BVDV1 or BVDV1-like virus. The acute vasculitis was accompanied by marked pulmonary and mesenteric edema and pleural effusion. In situ hybridization demonstrated abundant intracytoplasmic porcine circovirus type 2 (PCV2) nucleic acid in endothelial, smooth muscle-like, and inflammatory cells within and around affected arteries. The pigs with lymphohistiocytic and plasmacytic vasculitis had lesions of systemic PCVAD, including multisystemic lymphoplasmacytic and histiocytic or granulomatous inflammation. PCV2 nucleic acid was detected in renal tubule epithelial cells, mononuclear inflammatory cells, and rare endothelial cells in noninflamed vessels in multiple tissues of these animals. The 2 pigs with acute vasculitis had no PCV2-specific antibodies (or a low titer of), whereas the pigs with lymphohistiocytic and plasmacytic vasculitis developed high antibody titers against this virus. These observations suggest that (1) acute vasculitis observed in the current studies is directly caused by PCV2b, (2) chronic vasculitis may in part be mediated by the subsequent immune response, and (3) host factors and viral strain may both contribute to vasculitis in animals infected with PCV2b.

Experimental infection of white-tailed deer fawns (Odocoileus virginianus) with bovine viral diarrhea virus type-1 isolated from free-ranging white-tailed deer.

The objective of the current study was to elucidate the within-host dynamics of bovine viral diarrhea virus (BVDV) type-1 infection to better understand how this virus could be maintained in white-tailed deer (Odocoileus virginianus, WTD) populations. The BVDV type-1 used in this study was originally isolated from a free-ranging WTD in Indiana. Four fawns were intranasally inoculated with 2 ml BVDV type-1 strain 544 WTD at a 10(6) tissue culture infectious dose (TCID(50))/ml. Two fawns were inoculated with sham inoculum (negative controls). Animals were bled on days -7, 0, 1, 7, and 14 postinoculation (PID) for a complete blood count, chemistry panel, buffy coat (BC), real-time RT-PCR, and virus neutralization (VN). On days 7 and 14 PID, nasal and rectal swabs were obtained for RT-PCR and two of the virus-inoculated fawns and one of the negative controls fawns were euthanized. At necropsy, multiple samples were obtained for histopathology and in situ hybridization (ISH). Quantitative RT-PCR was performed on serum, BC, nasal, and rectal swabs. All animals tested negative for BVDV type 1 neutralizing antibodies on day 0 and animals in the control group remained seronegative throughout the study. No gross lesions were observed at necropsy. BVDV was isolated from lung and pooled lymph nodes from all BVDV-inoculated fawns on days 7 and 14 PID. Infected deer had lymphoid depletion, apoptosis, and lymphoid necrosis in the Peyer's patches and mesenteric lymph nodes. BVDV was detected in lymphoid tissues of infected animals by ISH. No lesions or virus were identified in control fawns. On day 7 PID, samples from two virus-inoculated fawns were positive for BVDV by virus isolation and RT-PCR from BC and nasal swab samples. One fawn was also positive on a rectal swab. Nasal and rectal swabs from all animals were negative on day 14. Results indicate that infection of WTD with BVDV is possible, and leads to histologic lesions in variety of tissues. In addition, virus shedding into the environment through feces and other secretions is likely.

Computed tomography imaging characteristics of canine nasal chondrosarcoma.

OBJECTIVES: To describe the CT appearance of canine nasal chondrosarcoma and compare calcification patterns with those described in humans. MATERIALS AND METHODS: CT examinations of dogs with biopsy-confirmed nasal chondrosarcoma were retrospectively reviewed and evaluated, including description of calcification patterns. RESULTS: Major findings are consistent with the previously described CT appearance of canine nasal tumours. Additionally, calcification patterns described as features of human nasal chondrosarcomas were also identified. CLINICAL SIGNIFICANCE: Findings may help prioritise differential lists of destructive nasal masses seen on CT in dogs.

IFN-γ induction by neutrophil-derived IL-17A homodimer augments pulmonary antibacterial defense.

The role of interleukin-17A (IL-17A) in host defense against Legionella pneumophila remains elusive. To address this issue, we used Il17a(-/-), Il17f(-/-), and Il17a/Il17f(-/-) mice on a C57Bl/6 (non-permissive) background and IL-17 neutralizing Abs in mice on an A/J (permissive) background. Higher bacterial (L. pneumophila) counts in the lung and blood along with reduced neutrophil recruitment were detected in Il17a(-/-), but not Il17f(-/-), mice. We found that neutrophils produce IL-17A homodimer (IL-17A) during L. pneumophila infection, and hematopoietic cell-derived IL-17A is known to be important for bacterial clearance. Thus, intratracheal administration of wild-type neutrophils or recombinant IL-17A restored bacterial clearance and neutrophil recruitment in Il17a(-/-) mice. Furthermore, neutrophil-depleted Rag2(-/-) and Rag2/Il-2rγ(-/-) mice exhibited increased bacterial burden, reduced neutrophil influx and IL-17A production in the lung. Recombinant IFN-γ administration in Il17a(-/-) mice augmented bacterial elimination, whereas IL-17A administration in Ifnγ(-/-) mice did not augment bacterial clearance. IFN-γ is produced by T cells, but not neutrophils or macrophages, suggesting that neutrophil-derived IL-17A induces IFN-γ in a paracrine fashion. Human pneumonic lungs and human neutrophils challenged with L. pneumophila exhibited increased numbers of IL-17A producing cells. These findings display a novel function of neutrophil-derived IL-17A in antibacterial defense via the induction of IFN-γ in a paracrine manner.

Sarcocystis spp. Infection in two Red Panda Cubs (Ailurus fulgens).

Two neonatal male red panda (Ailurus fulgens) littermates were submitted for necropsy examination. One animal was found dead with no prior signs of illness; the other had a brief history of laboured breathing. Post-mortem examination revealed disseminated protozoal infection. To further characterize the causative agent, transmission electron microscopy (TEM), immunohistochemistry (IHC), polymerase chain reaction (PCR) and amplification and nucleic acid sequencing were performed. IHC was negative for Toxoplasma gondii and Neospora caninum, but was positive for a Sarcocystis spp. TEM of cardiac muscle and lung revealed numerous intracellular apicomplexan protozoa within parasitophorous vacuoles. PCR and nucleic acid sequencing of partial 18S rRNA and the internal transcribed spacer (ITS)-1 region confirmed a Sarcocystis spp. that shared 99% sequence homology to Sarcocystis neurona and Sarcocystis dasypi. This represents the first report of sarcocystosis in red pandas. The histopathological, immunohistochemical, molecular and ultrastructural findings are supportive of vertical transmission resulting in fatal disseminated disease.

Feasibility of aqueous shunts for reduction of intraocular pressure in horses.

REASONS FOR PERFORMING STUDY: Based on the current literature, neither medical, surgical nor combination therapy adequately controls equine glaucoma for many horses. Aqueous shunts have been useful in other species to control glaucoma. OBJECTIVES: To determine whether aqueous shunts in normal equine eyes significantly reduce intraocular pressure (IOP) without causing vision threatening complications. STUDY DESIGN: Prospective experimental trial. METHODS: Aqueous shunts were placed in 7 normal eyes of 4 horses. The shunts were placed dorsotemporally. Examinations were initially performed daily for 7 days and after that every 3 days through 4 weeks after implantation. Horses were then subjected to euthanasia and globes enucleated for routine histological examination. The IOPs for each day post operatively were compared to the preoperative value (Day -1) using a Wilcoxon signed ranks test. Significance was set at P<0.05. RESULTS: The mean IOP preoperatively (20.7 ± 3.0 mmHg) was significantly higher than on any post operative day (P values ranged from 0.018 to 0.048). The aqueous shunts remained in situ for the entire study. Two eyes developed corneal ulcers that resolved. Shallow anterior chambers were noted in 2 eyes after shunt placement, which normalised after placement of full eye cup masks. Histologically, 7/7 eyes had fibrosis surrounding the implant. Minimal peripheral neovascularisation and neutrophilic keratitis were noted in 5/7 eyes. Corneal damage was scored as none in 3/7, mild in 2/7, moderate in 1/7 and marked in 1/7 eyes. CONCLUSIONS: After placement of aqueous shunts, a significant decrease in IOP was noted from preoperatively (Day -1) to Day 28 despite fibrosis surrounding the implants. No vision threatening complications were noted. POTENTIAL RELEVANCE: Aqueous shunts may represent a feasible therapeutic option for equine glaucoma. The results of this study suggest that further studies in glaucomatous horses would be warranted.

Enteric colonization by staphylococcus delphini in four ferret kits with diarrhoea.

Four, 1-to 4-week-old ferret kits were submitted to the Diagnostic Center for Population and Animal Health at Michigan State University for post-mortem examination. Grossly, multiple bowel loops in all ferret kits were distended by mucoid faecal material. Microscopically, there was no evidence of inflammation or notable alteration to the normal mucosal morphology. Gram-positive coccoid bacteria colonized variable segments of the small intestine. These bacteria were identified as Staphylococcus delphini by phenotypic and molecular analyses. Enzyme-linked immunosorbent assay for detection of Staphylococcus enterotoxins was positive and polymerase chain reaction detected the gene for Staphylococcus enterotoxin E in the isolates. The hypersecretory diarrhoea in these ferret kits may have been associated with colonization of the small intestine by S. delphini, cultures of which were shown in vitro to be potentially capable of producing enterotoxin E. The condition described in these ferrets is similar to 'sticky' kit syndrome in mink.

Coccidioidomycosis presenting as a heart base mass in two dogs.

Two atypical cases of canine coccidioidomycosis presenting as heart base masses are described. An echocardiogram performed in one of the two dogs revealed a large mass at the base of the heart and a computed tomography scan showed that the mass compressed the bronchi, left atrium, aorta and pulmonary arteries. A firm, white or pale yellow mass was found at the base of the heart at necropsy examination in both cases. Microscopical examination of the masses revealed severe, chronic, locally extensive granulomatous or pyogranulomatous inflammation with intralesional spherules consistent with Coccidioides spp. The diagnosis was further confirmed by immunohistochemistry and in-situ hybridization. Coccidioides spp. have been reported to cause pericarditis in dogs, but this is the first description of coccidioidomycosis mimicking a heart-based tumour in dogs.

Transitional carcinoma with extensive invasion of the bony orbit in a dog / Carcinoma transicional com invasão dos ossos da órbita em um cão

A 12-year-old male English Pointer was examined due to a soft-tissue swelling at the medial canthus of the right orbital region, which was causing facial deformity. The dog had epiphora, purulent nasal discharge, epistaxis, dyspnea, and progressive weight loss. An intraoral mass was observed near the right maxillary premolars. Neoplastic disease was diagnosed based on ancillary tests, which included blood work, skull and intraoral radiographs, ocular ultrasonography and computed tomography. Histopathology revealed transitional carcinoma involving the nasal and oral cavities, maxilla, bony orbit and retrobulbar space. Nasal tumors represent approximately 2% of all tumors diagnosed in this species. Transitional carcinoma is the second most common type of malignant epithelial tumor in the nasal sinuses. This case illustrates the extensive destruction of the soft and bony tissues of the face, including the bony orbit that this type of tumor can cause.

Um cão da raça Pointer Inglês, de 12 anos de idade, foi encaminhado para avaliação clínica por apresentar aumento de volume no canto temporal (medial) da órbita direita, o qual estava causando deformidade facial. O cão apresentava epífora, secreção nasal mucopurulenta, epistaxe, dispnésia e perda de peso progressiva. Uma massa foi observada na cavidade oral, próxima aos dentes pré-molares, do lado direito da face. Suspeitou-se de proliferação neoplásica baseando-se nos testes auxiliares, entre os quais: radiografia craniana e oral, ultrassonografia ocular e tomografia computadorizada. A análise histopatológica da massa revelou tratar-se de carcinoma transicional envolvendo as cavidades oral e nasal, os ossos da maxila e da órbita, além do espaço retrobulbar. Os tumores da cavidade nasal representam aproximadamente 2% de todos os tumores diagnosticados nessa espécie. O carcinoma transicional é o segundo tipo de neoplasia epitelial maligna mais comum nos seios nasais de cães. Este trabalho descreve a destruição extensiva dos tecidos moles e dos ossos da face e chama atenção para o fato de que esse tipo de neoplasia deve ser considerado nas doenças da órbita.

Toxoplasmosis sistémica en un canguro rojo (Macropus rufus) cautivo / Systemic toxoplamosis in a captive red kangaroo (Macropus rufus)

Los objetivos del estudio fueron presentar y documentar los hallazgos histopatológicos de toxoplasmosis sistémica en un canguro rojo (Macropus rufus) mantenido en cautiverio donde se describen los hallazgos macro y microscópicos encontrados y los análisis adicionales realizados. En el laboratorio de histopatología animal (Universidad de los Llanos) se recibieron muestras de tejidos fijados en formol tamponado, al 10% que procedían de un ejemplar macho de Macropus rufus, de ocho años de edad y 50 kg de peso corporal. Las muestras se procesaron mediante métodos rutinarios para microscopía óptica. Los cortes histológicos de 3-4 mm de grosor se colorearon con Hematoxilina-Eosina (H&E) y se realizó en algunos cortes la tinción de Ácido Periódico Schiff (PAS), PCR e IHQ. Al análisis histopatológico se encontró una toxoplasmosis sistémica asociada a quistes de protozoarios con inmunoreactividad positiva para T. gondii. La detección de T gondii en tejidos en formalina fue hecha usando dos ensayos de PCR que señalaban segmentos de ADN de diferentes secuencias repetitivas encontradas en T gondii y la IHQ confirmo lo hallado por PCR. Histopatológicamente se diagnosticó infección crónica por protozoarios eucoccideos de la familia Sarcocystidae. El diagnóstico etiológico fue de toxoplasmosis.

The objetives of this study were to present and document the hystopathologycal findings of systemic toxoplamosis in a captive red kangaroo (Macropus rufus) which described macro and microscopic findings of the hystopathological analysis. In the laboratory of animal histopathology (Universidad de los Llanos) formalin fixed tissue specimens were received, from a captive male Macropus rufus, who was eight years old and weighed 50 kg. The samples were processed by usual methods for optical microscopy. The histological sections of 3-4 mm thick were colored with Hematoxilin-Eosin (H&E) and then some samples stained with Periodic Acid Schiff (PAS), and processed by PCR and IHQ. Once the histopathological analysis was performed systemic toxoplasmosis was associated to protozoa cysts immunoreactives to T. gondii. The molecular detection of T. gondii in formalin fixed tissues was made using two PCR tests and confirmated by IHQ. Histopathologically a chronic infection by an eucoccideo protozoa from the Sarcocystidae family was diagnosed. The etiologic diagnosis was toxoplasmosis.

Listeric meningoencephalomyelitis in a cougar (Felis concolor): characterization by histopathologic, immunohistochemical, and molecular methods.

Listeria monocytogenes has been recognized as an important food-borne pathogen in animals. Records of the disease caused by this bacterium in large felids are, however, rare. The nervous form of listeriosis was diagnosed in a 12-year-old male cougar (Felis concolor) with a several-day history of neurologic disease characterized by excess salivation, head pressing, and circling that progressed to recumbency and death. Microscopically, the main alteration in the brain and spinal cord was a variably severe meningoencephalomyelitis composed mainly of mononuclear cell aggregates with fewer neutrophils. L. monocytogenes was isolated from the brain by microbiological culture, and L. monocytogenes antigen was detected in formalin-fixed, paraffin-embedded sections of brain and spinal cord by immunohistochemical analysis. On the basis of the nucleotide sequence of the 16S rRNA gene, the isolated strain was determined to be serotype 1/2a. Food-borne transmission of the bacterium was suspected, but food was not available for testing.