Protozoal meningoencephalitis and myelitis in 4 dogs associated with Trypanosoma cruzi infection.

American trypanosomiasis is caused by the zoonotic protozoa Trypanosoma cruzi and primarily results in heart disease. Organisms also infect the central nervous system (CNS). The Texas A&M University veterinary teaching hospital archive was searched for dogs with CNS disease with intralesional protozoal amastigotes. This study summarizes 4 cases of dogs with disseminated trypanosomiasis and CNS involvement confirmed by quantitative polymerase chain reaction (qPCR) with T. cruzi primers. Clinical signs included lethargy, respiratory distress, tetraparesis, and seizures. Central nervous system lesions included meningeal congestion (1/4), necrosis with hemorrhage in the spinal cord gray and white matter (2/4), and histiocytic meningoencephalitis (4/4), and meningomyelitis (2/4) with intralesional and intracellular protozoal. Genotyping identified 1 case of T. cruzi discrete typing unit (DTU) TcI and 2 cases as TcIV, both are common variants in the United States. Trypanosomiasis should be considered a differential diagnosis for dogs with CNS signs in T. cruzi-endemic areas.

Naegleria fowleri-associated meningoencephalitis in a cow in Southern Brazil-first molecular detection of N. fowleri in Brazil.

Naegleria fowleri, a free-living and thermophilic ameba, is the etiological agent of primary amebic meningoencephalitis (PAM). PAM is a rare and highly fatal neurologic disease in humans, and has been rarely documented in animal species. This report describes the pathological and etiological findings of a fatal case of N. fowleri-associated meningoencephalitis in a cow in Southern Brazil. Microscopic findings were consistent with severe, multifocal, hemorrhagic, and necrosuppurative meningoencephalitis associated with a large number of amebic trophozoites compatible with N. fowleri. Brain samples subjected to molecular assays generated a 315 bp fragment, which presented 99% identity with a N. fowleri sequence previously deposited in GenBank. This is the first study reporting the molecular detection of N. fowleri in a case of cattle meningoencephalitis in Latin America, and the obtained sequence represents the first GenBank deposit of N. fowleri identified in Brazil to this day. Additionally, the case reported is the second occurrence of N. fowleri-associated disease in the same city, drawing attention to the local importance of infection by this ameba and potential risk for human infections.

Diagnostic evaluation of fatal Balamuthia mandrillaris meningoencephalitis in a captive Bornean orangutan (Pongo pygmaeus) with identification of potential environmental source and evidence of chronic exposure.

A female Bornean orangutan (Pongo pygmaeus) aged 11 years and 6 months was examined by veterinarians after caretakers observed lethargy and facial grimacing. Within 72 h the primate had left-sided hemiparesis that worsened over the next week. An MRI revealed a focal right-sided cerebral mass suspected to be a neoplasm. Ten days after onset of clinical signs, the orangutan died. On postmortem exam, the medial right parietal lobe was replaced by a 7 × 4 × 3.5 cm focus of neuromalacia and hemorrhage that displaced the lateral ventricle and abutted the corpus callosum. Histopathology of the cerebral lesion revealed pyogranulomatous meningoencephalitis with intralesional amoeba trophozoites and rare cysts. Fresh parietal lobe was submitted to the Centers for Disease Control and Prevention lab for multiplex free-living amoebae real-time PCR and detected Balamuthia mandrillaris DNA at a high burden. Mitochondrial DNA was sequenced, and a 760-bp locus 19443F/20251R was compared to several human infections of B. mandrillaris and shown to be identical to the isolates from four human cases of encephalitis: 1998 in Australia, 1999 in California, 2000 in New York, and 2010 in Arizona. Indirect immunofluorescent antibody testing of stored serum samples indicated exposure to B. mandrillaris for at least 2 years prior to death. Within 1 week of the orangutan's death, water from the exhibit was analyzed and identified the presence of B. mandrillaris DNA, elucidating a possible source of exposure. B. mandrillaris, first reported in a mandrill in 1986, has since occurred in humans and animals and is now considered an important emerging pathogen.

First molecular characterization of Sarcocystis neurona causing meningoencephalitis in a domestic cat in Brazil.

Sarcocystis neurona is the main agent associated with equine protozoal myeloencephalitis (EPM). Apart from horses, S. neurona has been occasionally described causing neurologic disease in several other terrestrial animals as well as mortality in marine mammals. Herein, we describe the clinical, pathological, and molecular findings of a fatal case of S. neurona-associated meningoencephalitis in a domestic cat. The causing agent was analyzed by multilocus genotyping, confirming the presence of S. neurona DNA in the tissue samples of the affected animal. Significant molecular differences were found in relation to S. neurona isolates detected in other regions of the Americas. In addition, the parasite was identical to Sarcocystis sp. identified in opossum sporocysts in Brazil at molecular level, which suggests that transmission of. S. neurona in Brazil might involve variants of the parasite different from those found elsewhere in the Americas. Studies including more samples of S. neurona would be required to test this hypothesis, as well as to assess the impact of this diversity.

Meningoencephalitis due to Naegleria fowleri in cattle in southern Brazil / Meningoencefalite por Naegleria fowleri em um bovino no sul do Brasil

Abstract Naegleria fowleri is a free-living amoeba commonly found in the environment, mainly in fresh water and soil. This protozoon is occasionally involved in cases of fatal central nervous system disease in humans and other animal species. We describe here a case of meningoencephalitis due to Naegleria fowleri in cattle, in southern Brazil. A four-year-old Angus cow presented a clinical history of initial mild neurological signs that progressed to paddling movements, opisthotonus and lateral recumbency after five days. This animal had been kept in an irrigated rice stubble paddock. Grossly, the main lesions consisted of multiple areas of malacia in the right olfactory bulb, piriform lobes, hippocampus, frontal lobe cortex and fornix, along with severe thickening of the mesencephalon and rhombencephalon leptomeninges. Microscopically, severe multifocal necrosuppurative and hemorrhagic meningoencephalitis associated with a large quantity of amoebic trophozoites was present. The latter were confirmed to be Naegleria spp., through immunohistochemistry. Based on the strong congruence with the histopathological data of known cases reported in the literature, a probable association with Naegleria fowleri was established. To our knowledge, this is only the second report of Naegleria fowleri-associated meningoencephalitis in cattle in South America, and it is the first in southern Brazil.

Resumo Naegleria fowleri é uma ameba de vida livre, comumente encontrada no meio ambiente, principalmente em água doce e no solo. Este protozoário é ocasionalmente associado a casos fatais de doença do sistema nervoso central em seres humanos e espécies animais. No presente trabalho, um caso de meningoencefalite por Naegleria fowleri em um bovino na região sul do Brasil é descrito. Uma vaca Angus, de quatro anos de idade apresentou histórico clínico caracterizado inicialmente por sinais neurológicos leves que progrediram para movimentos de pedalagem, opistótono e decúbito lateral após cinco dias. Este animal era mantido em um piquete em resteva de arroz irrigado. Macroscopicamente, as principais lesões foram caracterizadas por múltiplas áreas de malacia no bulbo olfatório direito, lobos piriformes, hipocampo, córtex do lobo frontal e no fórnix, bem como acentuado espessamento das leptomeninges do mesencéfalo e rombencéfalo. Microscopicamente, meningoencefalite necrossupurativa e hemorrágica associada à grande número de trofozoítos amebianos foram observadas. Estes foram confirmados como Naegleria spp. através de imuno-histoquímica. Baseado na forte congruência apresentada entre os dados histopatológicos provenientes de casos conhecidos publicados na literatura, uma provável associação com Naegleria fowleri foi estabelecida. O presente trabalho trata-se do segundo relato de meningoencefalite associada à Naegleria fowleri em bovinos na América do Sul e o primeiro na região sul do Brasil.

Meningoencephalitis due to Naegleria fowleri in cattle in southern Brazil.

Naegleria fowleri is a free-living amoeba commonly found in the environment, mainly in fresh water and soil. This protozoon is occasionally involved in cases of fatal central nervous system disease in humans and other animal species. We describe here a case of meningoencephalitis due to Naegleria fowleri in cattle, in southern Brazil. A four-year-old Angus cow presented a clinical history of initial mild neurological signs that progressed to paddling movements, opisthotonus and lateral recumbency after five days. This animal had been kept in an irrigated rice stubble paddock. Grossly, the main lesions consisted of multiple areas of malacia in the right olfactory bulb, piriform lobes, hippocampus, frontal lobe cortex and fornix, along with severe thickening of the mesencephalon and rhombencephalon leptomeninges. Microscopically, severe multifocal necrosuppurative and hemorrhagic meningoencephalitis associated with a large quantity of amoebic trophozoites was present. The latter were confirmed to be Naegleria spp., through immunohistochemistry. Based on the strong congruence with the histopathological data of known cases reported in the literature, a probable association with Naegleria fowleri was established. To our knowledge, this is only the second report of Naegleria fowleri-associated meningoencephalitis in cattle in South America, and it is the first in southern Brazil.

Analysis of neurofilament concentration in healthy adult horses and utility in the diagnosis of equine protozoal myeloencephalitis and equine motor neuron disease.

Neurofilaments (NFs) are structural proteins of neurons that are released in significant quantities in the cerebrospinal fluid and blood as a result of neuronal degeneration or axonal damage. Therefore, NFs have potential as biomarkers for neurologic disorders. Neural degeneration increases with age and has the potential to confound the utility of NFs as biomarkers in the diagnosis of neurologic disorders. We investigated this relationship in horses with and without neurological diagnosis. While controlling for horse type (draft, pleasure, and racing), we evaluated the relationship between serum heavy-chain phosphorylated neurofilaments (pNF-H) and age, sex, and serum vitamin E concentrations. Serum pNF-H concentrations increased by 0.002 ng/ml for each year increase in age. There were significant differences in the serum pNF-H concentration among the type of activity performed by the horse. The highest serum pNF-H concentration was found in horses performing heavy work activity (racehorse) and with lower serum pNF-H concentration found among light (pleasure riding) and moderate (draft) activity. There was no significant association between the pNF-H concentration and sex or vitamin E concentration. Serum pNF-H concentration was elevated among horses afflicted with EMND and EPM when compared with control horses without evidence of neurologic disorders. Accordingly, serum pNF-H concentration can serve as a useful biomarker to complement the existing diagnostic work-up of horses suspected of having EPM or EMND.

Utility of C-reactive protein and serum amyloid A in the diagnosis of equine protozoal myeloencephalitis.

BACKGROUND: Accurate antemortem EPM diagnosis requires evidence of intrathecal antibody production. Some advocate the use of acute phase proteins in addition to serology, which alone results in substantial false positives. HYPOTHESIS/OBJECTIVES: The purpose of this study was to determine if serum C-reactive protein (CRP) or serum amyloid A (SAA) concentrations were elevated in cases of equine protozoal myeloencephalitis (EPM) compared to other neurological diseases. ANIMALS: 25 clinical cases of equine neurological disease: EPM (10), cervical vertebral stenotic myelopathy (CVSM) (10), neuroborreliosis (2), equine motor neuron disease (1), degenerative myelopathy (1), and leukoencephalomalacia (1). METHODS: Serum and CSF CRP and SAA were measured. Selection criteria included neurologic disease, antemortem diagnosis of EPM or CVSM, or postmortem diagnosis of EPM, CVSM, or other neurologic disease, and availability of serological results and archived samples for testing. RESULTS: Serum SAA and serum CRP levels were generally undetectable or low in horses with EPM (median CRP ≤0.1 mg/L, ≤0.1-14.4 mg/L; median SAA ≤0.1 mg/L, ≤0.1-6.11 mg/L) and CVSM (median CRP ≤0.1, ≤0.1-2.41 mg/L; median SAA ≤0.1mg/L, ≤0.1-13.88 mg/L). CSF CRP and SAA for horses with EPM (median CRP 3.35 mg/l, 0.19-13.43 mg/l; median SAA ≤0.1 mg/L, ≤0.1-2.4 mg/L) and CVSM (median CRP 4.015 mg/L, 0.16-9.62 mg/L; median SAA 0.62 mg/L, ≤0.1-2.91 mg/L) were also undetectable or low. Kruskal-Wallis test showed no statistically significant differences between serum CRP (P = .14), serum SAA (P = .79), spinal fluid CRP (P = .65), or spinal fluid SAA between horses with EPM and CVSM (P = .52). CONCLUSION: Neither SAA nor CRP in serum or CSF aid diagnosis of EPM.

Canine amoebic meningoencephalitis due to Balamuthia mandrillaris.

A 1-year-old Siberian Husky dog with acute-onset of seizures, recumbency, paddling, and muscular fasciculations was autopsied. A locally extensive hemorrhagic and malacic focus was noted in the right cerebral frontal cortex, and severe necrotizing and hemorrhagic, neutrophilic meningoencephalitis was diagnosed microscopically. Amoebic trophozoites and cysts were identified within the affected cerebral parenchyma and confirmed by indirect immunofluorescence assay and real-time PCR as Balamuthia mandrillaris. B. mandrillaris is found in soil and water and the infection has been reported in both immunocompromised and immunocompetent humans and rarely in the dog.

Meningoencephalitis caused by pathogenic Sarcocystis species in a naturally infected sheep in Turkey.

A 3-year-old sheep was examined after an acute onset of hind limb paralysis and ataxia. At necropsy, central nervous system, pulmonary and intestinal hyperaemia and ecchymoses in the aortic arch were observed. Main microscopic lesions were confined to the heart, cerebrum and cerebellum. There were a multifocal mild myocarditis and nonsuppurative meningoencephalitis together with protozoal cysts in the heart and the brain. Protozoal cystic structures were observed within many of the myocardial fibers as well as in the cerebrum and cerebellum. Using light microscopy it could not be morphologically determined whether these organisms were Toxoplasma (T.) gondii or Neospora (N.) caninum. Additional diagnostic methods like immunohistochemistry and polymerase chain reaction provided differentiation of Sarcocystis from T. gondii and N. caninum. Transmission electron microscopy demonstrated characteristic features of Sarcocystis sp. as previously described. This is the first confirmed diagnosis of Sarcocystis sp. in the central nervous system of a sheep from Turkey.

A case of canine neurological leishmaniasis.

In this study we describe a case of neurological leishmaniasis in a dog, reported in an endemic area, with signs of multifocal intracranial lesions involving the vestibular system and the cerebellum. Serum biochemistry revealed a decrease of albumin-globulin ratio caused by an increase of alfa2-, beta-, and gamma- globulin, while antibody titers were highly positive for Leishmania spp. Magnetic resonance images of the brain were consistent with a granulomatous in ammation/infection. Cerebrospinal fluid revealed a marked mononuclear pleocytosis and was positive to the Pandy Test, as well as to a Leishmania spp. -speci c polymerase chain reaction (PCR) assay. Sequencing of the PCR products revealed the highest similarity with several strains belonging to the Leishmania infantum / Leishmania chagasi group. Clinical response to treatment for systemic leishmaniasis was supportive of diagnosis. This report focuses on an atypical form of canine leishmaniasis and suggests that in endemic geographic areas leishmaniasis has to be considered for di erential diagnosis in neurological manifestations. Also, cerebrospinal liquor should always be tested when neurological symptoms are present in order to di erentiate neurological leishmaniasis from unrelated neurological signs in Leishmania positive patients.

Meningoencephalitis due to the amoeboflagellate Naegleria fowleri in ruminants in Algeria.

Primary amoebic meningoencephalitis (PAM) is a fatal infection in most cases, caused by the amoeba flagellate Naegleria fowleri. This report describes the first cases of PAM in Algeria, in a cow and a ewe from Batna, north-eastern Algeria. The death of both ruminants occurred a week after the first clinical manifestations. The cerebrospinal fluid, after staining with May-Grünwald-Giemsa, showed the presence of amoebae cells. Histological sections revealed numerous amoebae in all parts of the brain. The presence of N. fowleri was confirmed using a species-specific real-time PCR in histological tissue sections. The two PAM cases were reported during the hot season, and the source of infection is very likely the water where the cattle came to drink. Particular attention should be focused on this type of infection in aquatic environments when the temperature is high and preventive measures must be taken to avoid the proliferation of N. fowleri.

Trichobilharzia regenti (Schistosomatidae): 3D imaging techniques in characterization of larval migration through the CNS of vertebrates.

Migration of parasitic worms through the host tissues, which may occasionally result in fatal damage to the internal organs, represents one of the major risks associated with helminthoses. In order to track the parasites, traditionally used 2D imaging techniques such as histology or squash preparation do not always provide sufficient data to describe worm location/behavior in the host. On the other hand, 3D imaging methods are widely used in cell biology, medical radiology, osteology or cancer research, but their use in parasitological research is currently occasional. Thus, we aimed at the evaluation of suitability of selected 3D methods to monitor migration of the neuropathogenic avian schistosome Trichobilharzia regenti in extracted spinal cord of experimental vertebrate hosts. All investigated methods, two of them based on tracking of fluorescently stained larvae with or without previous chemical clearing of tissue and one based on X-ray micro-CT, exhibit certain limits for in vivo observation. Nevertheless, our study shows that the tested methods as ultramicroscopy (used for the first time in parasitology) and micro-CT represent promising tool for precise analyzing of parasite larvae in the CNS. Synthesis of these 3D imaging techniques can provide more comprehensive look at the course of infection, host immune response and pathology caused by migrating parasites within entire tissue samples, which would not be possible with traditional approaches.

Amoebic meningoencephalitis and disseminated infection caused by Balamuthia mandrillaris in a Western lowland gorilla (Gorilla gorilla gorilla).

CASE DESCRIPTION A 22-year-old male gorilla (Gorilla gorilla gorilla) housed in a zoo was evaluated for signs of lethargy, head-holding, and cervical stiffness followed by development of neurologic abnormalities including signs of depression, lip droop, and tremors. CLINICAL FINDINGS Physical examination under general anesthesia revealed a tooth root abscess and suboptimal body condition. A CBC and serum biochemical analysis revealed mild anemia, neutrophilia and eosinopenia consistent with a stress leukogram, and signs consistent with dehydration. Subsequent CSF analysis revealed lymphocytic pleocytosis and markedly increased total protein concentration. TREATMENT AND OUTCOME Despite treatment with antimicrobials, steroids, and additional supportive care measures, the gorilla's condition progressed to an obtunded mentation with grand mal seizures over the course of 10 days. Therefore, the animal was euthanized and necropsy was performed. Multifocal areas of malacia and hemorrhage were scattered throughout the brain; on histologic examination, these areas consisted of necrosis and hemorrhage associated with mixed inflammation, vascular necrosis, and intralesional amoebic trophozoites. Tan foci were also present in the kidneys and pancreas. Immunohistochemical testing positively labeled free-living amoebae within the brain, kidneys, eyes, pancreas, heart, and pulmonary capillaries. Subsequent PCR assay of CSF and frozen kidney samples identified the organism as Balamuthia mandrillaris, confirming a diagnosis of amoebic meningoencephalitis. CLINICAL RELEVANCE Infection with B mandrillaris has been reported to account for 2.8% of captive gorilla deaths in North America over the past 19 years. Clinicians working with gorillas should have a high index of suspicion for this diagnosis when evaluating and treating animals with signs of centrally localized neurologic disease.

Neurotropic T-cell-rich B-cell lymphoma in a 14-year-old Morgan gelding.

A 14-year-old Morgan gelding was presented for progressive weakness and muscle atrophy. The horse was initially diagnosed with equine protozoal myelitis based on history, physical examination, and laboratory diagnostics. Despite therapy, the horse declined clinically and was euthanized. Necropsy revealed a rare form of neurotropic lymphoma, described in this report.

Lymphome de cellules-B riches en cellules-T neurotropes chez un hongre Morgan âgé de 14 ans. Un hongre Morgan âgé de 14 ans a été présenté pour une faiblesse progressive et une atrophie musculaire. On a d'abord diagnostiqué la myélite protozoaire équine chez le cheval en se basant sur l'anamnèse, l'examen physique et le diagnostic en laboratoire. Malgré la thérapie, l'état clinique du cheval s'est détérioré et il a été euthanasié. La nécropsie a révélé une forme rare de lymphome neutropique, qui est décrite dans ce rapport.(Traduit par Isabelle Vallières).

Systemic Scuticociliatosis (Philasterides dicentrarchi) in sharks.

Scuticociliatosis is an economically important, frequently fatal disease of marine fish in aquaculture, caused by histophagous ciliated protozoa in the subclass Scuticociliatida of the phylum Ciliophora. A rapidly lethal systemic scuticociliate infection is described that affected aquarium-captive zebra sharks (Stegostoma fasciatum), Port Jackson sharks (Heterodontus portusjacksoni), and a Japanese horn shark (Heterodontus japonicus). Animals died unexpectedly or after a brief period of lethargy or behavioral abnormality. Gross findings included necrohemorrhagic hepatitis and increased volumes of celomic fluid. Histologically, 1 or more of a triad of necrotizing hepatitis, necrotizing meningoencephalitis, and thrombosing branchitis were seen in all cases, with necrotizing vasculitis or intravascular fibrinocellular thrombi. Lesions contained variably abundant invading ciliated protozoa. Molecular identification by polymerase chain reaction from formalin-fixed tissues identified these as the scuticociliate Philasterides dicentrarchi (syn. Miamiensis avidus), a novel and potentially emergent pathogen in sharks.

Canine cerebral leishmaniasis: potential role of matrix metalloproteinase-2 in the development of neurological disease.

Matrix metalloproteinases (MMPs) are a group of calcium- and zinc-dependent endopeptidases that are involved in maintaining the extracellular matrix. MMP-2 and MMP-9 are thought to be related to the disruption of the blood-brain-barrier (BBB) by their ability to cleave type IV collagen, the main component of the basal membrane. To establish the presence of MMP-2 and MMP-9 in the pathogenesis of canine cerebral leishmaniasis, we examined the levels of these metalloproteinases in the cerebrospinal fluid (CSF) and serum of dogs with visceral leishmaniasis and neurological symptoms (n=16) and in the CSF and serum of uninfected healthy dogs (n=10) using zymography. In the CSF of dogs with cerebral leishmaniasis there was a massive presence of active MMP-2, whereas only the levels of both proMMP-2 and proMMP-9 were elevated in the serum. Although the detected MMP activity in the CSF might merely be related to CNS inflammation, these enzymes may also play a collaborative role in the disease progression. Both MMP-2 and MMP-9 are known to target critical constituents of the BBB, and once activated, they may promote cerebral barrier breakdown, allowing the entrance of inflammatory cells and proteins within the nervous system milieu.

Meningoencephalitis caused by Naegleria fowleri in cattle of northeast Brazil.

Primary amoebic meningoencephalitis is an acute disease of the central nervous system of humans and animals caused by Naegleria fowleri. This report describes a case of meningoencephalitis caused by N. fowleri in a crossbred, one-year-old bovine with progressive neurological signs. At necropsy there was thickening of the meninges and multifocal areas of malacia in the thalamus, caudal and rostral colliculi, parietal and occipital cortex, and cerebellum. Histologically there was multifocal necrosupurative meningoencephalitis associated with areas of malacia containing amoebic trophozoites. Immunohistochemistry of the brain was positive for N. fowleri. The disease should be included among the differential diagnosis of diseases of the central nervous system in cattle in areas where animals are exposed to hot, stagnant water.

Fatal Balamuthia mandrillaris infection in a gorilla - first case of balamuthiasis in Germany.

BACKGROUND: A 12-year-old female western lowland gorilla died in a zoological garden in Germany after exhibiting general neurological signs. METHODS: Balamuthia mandrillaris was identified as causative agent by indirect immunofluorescent staining of brain sections and confirmed by PCR and respective sequencing. RESULTS: The animal suffered from a chronic progressive necrotizing amebic meningoencephalitis. CONCLUSION: This is the first case of Balamuthia amebic encephalitis in Germany.