Publications by authors named "Luciana Sonne"

33 Publications

Pathological and immunohistochemical aspects of acute megakaryoblastic leukaemia in a cat - Short communication.

Acta Vet Hung 2021 Jul 2. Epub 2021 Jul 2.

1Department of Veterinary Pathology, Federal University of Rio Grande do Sul (UFRGS), Av. Bento Gonçalves 9090, Porto Alegre, Rio Grande do Sul, 91540-000, Brazil.

An adult, mixed-breed, feline leukaemia virus (FeLV-) positive female cat was presented with mucosal jaundice and a history of anorexia and constipation for three days. Physical examination revealed splenomegaly, cachexia, and dehydration. Humane euthanasia was conducted, followed by postmortem examination. Grossly, the cat was icteric, and presented hepatomegaly with multifocal white spots and splenomegaly. Histologically, the bone marrow was nearly completely replaced by a proliferation of megakaryocytes and megakaryoblasts, and there was a proliferation of fibrous connective tissue. Similar neoplastic proliferation was observed infiltrating the liver, lymph nodes, spleen, kidney, skeletal muscle, and lungs. Immunohistochemistry was performed for von Willebrand Factor (VWF), CD79α, CD3, feline immunodeficiency virus, FeLV, and CD61. Marked cytoplasmic labelling was observed in the neoplastic cells for FeLV, VWF and CD61, corroborating the diagnosis of acute megakaryoblastic leukaemia.
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http://dx.doi.org/10.1556/004.2021.00025DOI Listing
July 2021

Rangelia vitalii in free-living crab-eating foxes (Cerdocyon thous) in Uruguay.

Ticks Tick Borne Dis 2021 Jun 4;12(5):101765. Epub 2021 Jun 4.

Setor de Patologia Veterinária, Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária (FAVET), Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brazil.

Rangelia vitalii is a protozoan parasite that causes a hemorrhagic and hemolytic disease in dogs known as rangeliosis. Current reports of the disease are concentrated in the southern and southeastern regions of Brazil, as well as in Uruguay, Argentina, and Paraguay, and mainly concern domestic dogs. South American wild canids, such as the crab-eating fox (Cerdocyon thous), the pampas fox (Lycalopex gymnocercus), and the maned wolf (Chrysocyon brachyurus) may also be affected, although existing reports are restricted to Brazil. The present study aimed to detect R. vitalii parasitism in the Uruguayan wild fox population. DNA extracted from the blood and/or spleen samples of road-killed C. thous and L. gymnocercus found in northern Uruguay were subjected to polymerase chain reaction (PCR) to amplify a 551-bp fragment of the Rangelia 18S rRNA gene. A total of 62 wild canids, including 38 C. thous and 24L. gymnocercus, were analyzed. Five crab-eating fox samples (13.2%) were positive for R. vitalii, with 99.5-100% identity between the sequences. All samples from pampas fox tested negative for R. vitalii. When compared with the R. vitalii sequences available in GenBank, a similarity of 98.9-100% was revealed. Molecular analysis results suggest that R. vitalii is circulating in the crab-eating fox population in Uruguay; however, its veterinary relevance for these foxes remains unknown.
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http://dx.doi.org/10.1016/j.ttbdis.2021.101765DOI Listing
June 2021

Comparison of immunohistochemical profiles of ovarian germ cells in dysgerminomas of a captive maned wolf and domestic dogs.

J Vet Diagn Invest 2021 Jul 28;33(4):772-776. Epub 2021 May 28.

Setor de Patologia Veterinária, Departamento de Patologia Clínica Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil.

We characterized the immunohistochemical expression profiles of dysgerminomas from a 16-y-old maned wolf and 13 domestic dogs using the following biomarkers: Sal-like protein 4 (SALL4), octamer-binding transcription factor 3/4 (OCT3/4), placental alkaline phosphatase (PLAP), c-kit, and vimentin. The maned wolf had nonspecific and long-standing clinical signs of lethargy, anorexia, and weight loss, and was euthanized because of poor prognosis. At autopsy, the left ovary was effaced by a 12 × 8 × 6 cm mass, comprised of anaplastic cells with a mitotic count of 20 mitoses in 10 high power fields. Dysgerminomas from 7 of 13 domestic dogs had nuclear expression of SALL4. Dysgerminomas from the maned wolf and 2 domestic dogs had both nuclear and cytoplasmic expression of SALL4. Cytoplasmic expression of PLAP and OCT3/4 was present in dysgerminomas from the maned wolf and 3 (PLAP) or 4 (OCT3/4) domestic dogs. All dysgerminomas expressed vimentin. Membranous c-kit expression was rare in the dysgerminoma from the maned wolf, and variable in dysgerminomas from 4 domestic dogs. A dysgerminoma from a domestic dog had cytoplasmic expression of c-kit. SALL4 is a useful marker to confirm germ cell origin of dysgerminoma in canids.
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http://dx.doi.org/10.1177/10406387211019959DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8229830PMC
July 2021

Enteric organisms detected in feces of dogs with bloody diarrhea: 45 cases.

Top Companion Anim Med 2021 May 24:100549. Epub 2021 May 24.

Veterinary School. Universidade Federal de Minas Gerais (UFMG). Av. Antônio Carlos, 6627. Belo Horizonte, MG, Brazil, CEP 31.270-901. Electronic address:

Bloody diarrhea is a common condition in dogs, but studies evaluating the enteropathogens involved specifically in adult dogs are scarce. In the present study, stool samples from 45 adult dogs with bloody diarrhea were evaluated for the four enteric organisms mainly reported in these cases: canine parvovirus type 2 (CPV-2), Clostridioides difficile, Clostridium perfringens, and Salmonella spp. In addition, the samples were also tested for coronavirus, rotavirus, Giardia spp., and Escherichia coli pathotypes to provide a better understanding of possible co-occurrence. Vaccination status, diet, and clinical outcome were also obtained when available. CPV-2b was identified in 17 dogs (37.8%), being the most frequent cause of bloody diarrhea, including completely vaccinated adult dogs. Toxigenic C. difficile and C. perfringens netF were detected in six (13.3%) and five (11.1%) dogs, in some cases in a co-occurrence with other enteric organisms. Three fatal cases of salmonellosis were identified in dogs fed a raw meat-based diet, raising the risks associated with this increasing practice.
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http://dx.doi.org/10.1016/j.tcam.2021.100549DOI Listing
May 2021

Rangelia vitalii molecular and histological quantification in tissues comparing crab-eating foxes (Cerdocyon thous) and domestic dogs.

Ticks Tick Borne Dis 2021 May 2;12(5):101731. Epub 2021 May 2.

Setor de Patologia Veterinária, Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária (FAVET), Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brazil.

Rangeliosis is a condition transmitted by the tick Amblyomma aureolatum and caused by the protozoan parasite Rangelia vitalii in canids. In domestic dogs, the disease causes a severe hemolytic disease, while in wild canids the piroplasm is often detected without any clinical abnormality. This study aimed to detect and quantify the number of copies of the R. vitalii Hsp70 gene (indirect parasite burden) in several organs of domestic and South American wild canids (Cerdocyon thous and Lycalopex gymnocercus) to elucidate distinct clinical presentations of rangeliosis in these species. A total of seven domestic dogs that died due to rangeliosis and 38 wild foxes were initially included, with all dogs presenting histological and molecular features of rangeliosis, while eight C. thous were positive at the molecular analysis for R. vitalii. Fragments of 22 organs collected from domestic (n = 7) and wild foxes (n = 8) were employed for histological and molecular quantification using real-time polymerase chain reaction aiming at the Hsp70 gene. Histologically, parasitophorous vacuoles were constantly detected in the dogs, while these were detected only in two C. thous. Parasitic burden was significantly higher in the digestive, cardiorespiratory, endocrine, genitourinary, and skeletal-muscle systems of domestic dogs when compared to wild foxes. In the hematopoietic system of wild canids, some organs, such as the lymph nodes and tonsils, presented significantly lower amounts of R. vitalii, while other organs (spleen, bone marrow, and blood) had results similar to those of domestic dogs. Additionally, the central nervous system of both domestic and wild canids presented a similar quantity of R. vitalii. The etiological agent is possibly maintained through an asexual reproductive process (merogony) in both domestic and wild species. Nonetheless, a limited or short-duration schizogony phase occurs in C. thous, which would designate this species as a possible reservoir host for the agent. Dogs, in contrast, would most likely act as accidental hosts, presenting a severe and more pathogenic schizogony phase, resulting in characteristic clinical and pathological rangeliosis.
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http://dx.doi.org/10.1016/j.ttbdis.2021.101731DOI Listing
May 2021

Pathological and Immunohistochemical Characterization of Thyroid Neoplasms in Cats.

J Comp Pathol 2021 Apr 2;184:44-55. Epub 2021 Mar 2.

Department of Veterinary Pathology, College of Veterinary Medicine, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil.

There are few pathological and immunohistochemical descriptions of thyroid neoplasms in cats. We investigated neoplasms of the thyroid in 31 cats (average age 15.6 years) in Southern Brazil. Most (96.8%) of these cases were classified as follicular adenomas and were predominantly unilateral and multinodular. Histologically, macrofollicular adenomas were the most common type observed. Carcinomas represented 3.2% of the investigated tumours. By immunohistochemical labelling, thyroglobulin was expressed more commonly than paired box gene 8 or thyroid transcription factor 1 in the follicular adenomas. One carcinoma was immunopositive for thyroglobulin, pancytokeratin, chromogranin A and synaptophysin, suggestive of a calcitonin-negative neuroendocrine carcinoma.
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http://dx.doi.org/10.1016/j.jcpa.2021.01.013DOI Listing
April 2021

Causes of death in beef cattle in southern Brazil.

J Vet Diagn Invest 2021 Jul 9;33(4):677-683. Epub 2021 Apr 9.

Faculdade de Veterinária, Setor de Patologia Veterinária, Universidade Federal do Rio Grande do Sul, Rio Grande do Sul, Brazil.

We determined the prevalence of diseases and pathogens associated with mortality in beef cattle in the State of Rio Grande do Sul, Brazil, based on pathology laboratory submissions. Postmortem examinations were conducted on 1,277 beef cattle that died between 2008 and 2018. Information regarding age, time of the year, breed, and regional location were analyzed statistically. Most cattle were from the surrounding region of Porto Alegre, and 78.7% of the analyzed cases had diagnostic value. The diagnostic category with most cases was infectious and/or parasitic diseases (60%), followed by toxic and toxicoinfectious (25%). Most cases occurred in the fall. Major disease conditions identified included hemoprotozoal infection (18.2%), rabies (8.2%), and plant intoxications by spp. (8.5%) and (4.6%). Hemoprotozoal infection occurred at a higher frequency in young cattle, mainly in animals up to 1 y old. Intoxication by spp. was more frequent in cattle 2-3 y old.
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http://dx.doi.org/10.1177/10406387211007952DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8229822PMC
July 2021

Relationship Between Clinicopathological Features and Prognosis in Appendicular Osteosarcoma in Dogs.

J Comp Pathol 2020 Oct 6;180:91-99. Epub 2020 Oct 6.

Setor de Patologia Veterinária, Brazil.

Osteosarcoma is the most common primary bone neoplasm in dogs and often involves the appendicular skeleton. We report the clinicopathological and prognostic features of appendicular osteosarcomas diagnosed in 153 dogs from 2008 to 2018. The survival data for 22 dogs that underwent surgery and chemotherapy were statistically compared with the clinical, histopathological and immunohistochemical data for correlation with prognosis. The affected dogs had a mean age of 9.1 years and a mean body weight of 33.4 kg. No sex predilection was detected, although the incidence was slightly higher in females (52%). Large mixed and purebred dogs were most commonly affected. Long bones were affected in 124 cases (92.5%; 124/134). The extremities affected within long bones were the metaphysis of the proximal humerus (29%), distal radius (17.7%), distal femur (15.3%) and proximal tibia (7.2%). Histologically, osteoblastic osteosarcoma was most commonly detected (56.9%), followed by chondroblastic (13.7%), telangiectatic (11.8%), fibroblastic (7.8%), giant cell-rich (5.2%) and poorly differentiated neoplasms (4.6%). Regional lymph nodes were available for analysis in 28 out of 70 excisional biopsies from amputated entire limbs. Metastases to regional lymph nodes were observed in 14.3% (4/28) of these cases. Distant metastases were found in 75% (39/52) of cases in which diagnostic imaging or necropsy was carried out. The lungs were the most common site of metastasis (87.2%; 34/39 cases). Weight, age, sex, affected limb or bone, histological classification, mitotic count or histological grade did not influence survival (P >0.05). Neoplasms located at proximal extremities (n = 7) of affected limbs had a tendency for a poorer prognosis than those at distal extremities (n = 15) (P = 0.06). For these cases, the 1-year survival rate was 14.3% compared with 40% for neoplasms at distal extremities. There was no significant difference in the survival rate when amputation (n = 16) or limb preservation (n = 6) techniques were employed with chemotherapy (P = 0.20). The survival period of dogs that had undergone limb amputation, or limb preservation surgery, and chemotherapy was 73-1,185 days, with a mean and median of 376 and 256 days, respectively. Mild to marked cytoplasmic immunolabelling of osteopontin was found in all cases but the intensity (P = 0.66) and percentage of neoplastic cells labelled (P = 0.49) did not influence survival.
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http://dx.doi.org/10.1016/j.jcpa.2020.09.003DOI Listing
October 2020

Feline giant-cell pleomorphic sarcoma: cytologic, histologic and immunohistochemical characterization.

J Feline Med Surg 2021 Aug 18;23(8):738-744. Epub 2020 Nov 18.

Department of Veterinary Pathology, College of Veterinary Medicine, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.

Objectives: This study aimed to characterize the cytologic, pathologic and immunohistochemical (IHC) aspects of feline giant-cell sarcoma.

Methods: Biopsy and necropsy reports from the Department of Veterinary Pathology were retrieved, and 13 cases of pleomorphic sarcoma (PS) were selected according to the established epidemiologic, pathologic and IHC criteria. All samples were fixed in 10% formalin, routinely processed for histology, and stained with hematoxylin and eosin. Samples also underwent IHC testing for vimentin, ionized calcium-binding adaptor molecule 1 (Iba-1), desmin, actin and S-100.

Results: The mean age of the affected cats was 9.5 years, and females were over-represented. Most neoplasms were observed in the flank, lateral thorax, limbs and interscapular region, and were >2 cm in diameter. Cytology analysis revealed highly cellular preparations with three distinct populations (spindle cells, small round cells and multinucleated giant cells) in a dense eosinophilic stroma. Histologically, PS was composed of a combination of these three populations. IHC labeling for vimentin and Iba-1 was strongly positive for spindle cells and multinucleated giant cells, respectively. Desmin/actin showed variable labeling among the samples. S-100 was negative in all samples.

Conclusions And Relevance: PS is a neoplasm of mesenchymal origin, also known as malignant fibrous histiocytoma. The predominant subtype in this study that affected the cats was the giant-cell type, characterized by the presence of multinucleated giant cells among spindle-shaped cells. These findings are similar to those described in human patients; thus, a comparison between the neoplasms seen in these species might be useful, and the knowledge of biologic behavior and overall treatment approach for humans could be extrapolated to cats.
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http://dx.doi.org/10.1177/1098612X20972667DOI Listing
August 2021

Fatal disseminated toxoplasmosis in a brown-throated sloth (Bradypus variegatus) from Northern Brazil - Case report.

Acta Vet Hung 2020 09 30;68(3):285-288. Epub 2020 Sep 30.

4Universidade Federal de Mato Grosso do Sul (UFMS), Campo Grande, Brazil.

The clinical and pathological findings of a case of fatal disseminated toxoplasmosis in a captive brown-throated sloth (Bradypus variegatus) from the northern region of Brazil are reported. Clinical signs were nonspecific and included apathy, prostration, dyspnoea, and loss of appetite. Treatment with penicillin was attempted, but the animal died within five days of the onset of clinical signs. Microscopically, there was acute inflammation in the liver, spleen, and lungs associated with necrosis and a few cysts and extracytoplasmic tachyzoites, with a morphology compatible with Toxoplasma gondii. Tissue sections were submitted for immunohistochemistry that confirmed T. gondii as the aetiological agent. To the authors' knowledge, this is the first report of toxoplasmosis in B. variegatus.
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http://dx.doi.org/10.1556/004.2020.00049DOI Listing
September 2020

What is your diagnosis? Ventral abdominal mammary mass in a bitch.

Vet Clin Pathol 2020 Dec 20;49(4):672-674. Epub 2020 Oct 20.

Setor de Patologia Veterinária, Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária (FAVET), Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.

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http://dx.doi.org/10.1111/vcp.12904DOI Listing
December 2020

Pathological, immunohistochemical, and molecular findings of equine protozoal myeloencephalitis due to Sarcocystis neurona infection in Brazilian horses.

Trop Anim Health Prod 2020 Nov 4;52(6):3809-3817. Epub 2020 Oct 4.

Setor de Patologia Veterinária, Universidade Federal do Rio Grande do Sul (UFRGS), Avenida Bento Gonçalves 9090, Prédio 42.505, Bairro Agronomia, Porto Alegre, Rio Grande do Sul, 91540-000, Brazil.

Equine protozoal myeloencephalitis (EPM) is an important neurologic disease of horses in the American continent caused by Sarcocystis neurona and Neospora hughesi infection. This study describes the pathological, immunohistochemical, and molecular findings of fatal cases of EPM in southern Brazil. A review was performed on a total of 13 cases compatible with EPM, which were diagnosed by postmortem examination in the period of 2010-2017. Epidemiological information was obtained from necropsy reports. Gross and histological lesions were characterized, and cases were subjected to immunohistochemistry anti-Sarcocystis neurona, Toxoplasma gondii, and Neospora spp. Molecular search was performed using ITS-1 gene PCRs. Microscopic lesions were multifocal in all cases, and more frequently observed in the spinal cord segments and in the rhombencephalon. Intralesional protozoans were histologically detected in five horses, while a positive immunostaining for S. neurona was observed in eleven cases (11/13). Through molecular techniques, six positive cases for the ITS-1 gene were detected, and obtained sequences presented highest similarity with S. neurona. EPM due to S. neurona infection represents an important neurologic disease of horses in Brazil and this disease should be considered as a main differential diagnosis in horses presenting neurologic signs.
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http://dx.doi.org/10.1007/s11250-020-02419-yDOI Listing
November 2020

Mycobacterium tuberculosis var. tuberculosis infection in two captive black capuchins (Sapajus nigritus) in Southern Brazil.

Braz J Microbiol 2020 Dec 22;51(4):2169-2173. Epub 2020 Jul 22.

Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gonçalves 9090, Rio Grande do Sul, 91540-000, Porto Alegre, Brazil.

Tuberculosis is a common zooanthroponosis in humans with a high incidence in Brazil, but it may also affect non-human primates (NHPs), of which Old World primates are most commonly involved. Nonetheless, its occurrence in New World primates is unknown, and therefore, this study aimed to describe the infection by Mycobacterium tuberculosis variant tuberculosis in two captive black capuchin monkeys (Sapajus nigritus) in Southern Brazil. The primates were housed in the same enclosure, wherein close contact with humans frequently occurred, and presented apathy, anorexia, and death in a clinical course of 15 days to 2 months. At the necropsy, the tracheobronchial lymph nodes were markedly enlarged and firm to hard and on the cut surface had a caseous aspect. The lungs exhibited two injury patterns: multifocal and disseminated. Microscopically, the lungs exhibited multifocal to coalescing necrotic granulomas and non-necrotic granulomas, with multiple acid-fast bacilli within the cytoplasm of epithelioid macrophages and multinucleated giant cells. Bacilli were also labeled upon immunohistochemistry (IHC) for Mycobacterium tuberculosis. Microbiological culture of lung samples from both cases yielded colonies compatible with M. tuberculosis. The isolates were identified as M. tuberculosis var. tuberculosis through polymerase chain reaction (PCR). Although tuberculosis is poorly described in New World primates, M. tuberculosis var. tuberculosis may cause a highly contagious and progressive disease with high mortality in black capuchin monkeys (S. nigritus).
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http://dx.doi.org/10.1007/s42770-020-00339-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7688760PMC
December 2020

Ophthalmopathologic characterization of multicentric or metastatic neoplasms with an extraocular origin in dogs and cats.

Vet Ophthalmol 2020 Sep 20;23(5):814-827. Epub 2020 Jul 20.

Department of Clinical Pathology, Faculty of Veterinary, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil.

Objective: To characterize the frequency and distribution of secondary ocular neoplasms in dogs and cats identified during necropsy.

Methods: A retrospective analysis of necropsy records of dogs and cats was conducted, and cases with metastatic/multicentric neoplasms with an extraocular origin, involving the eyes, were selected.

Results: From January 2015 to January 2019, we identified a total of 233 dogs and 100 cats with metastatic disease. Of these, 11.6% (27/233) of the dogs and 13% (13/100) of the cats had ocular metastases. Lymphoma was the most common multicentric neoplasm involving the eyes of both species. In dogs, these neoplasms occurred bilaterally, predominantly in the anterior uvea, and were diffuse large B-cell, T-lymphoblastic, peripheral T-cell not otherwise specified, and lymphocytic B-cell lymphomas. In cats, feline leukemia virus (FeLV)-associated T-cell lymphoma was the most common. Mammary carcinoma was the second most common ocular metastatic neoplasm in bitches, with a predominantly unilateral involvement of the uveal tract. In cats, following lymphoma, pulmonary and squamous cell carcinomas were the most common multicentric/metastatic neoplasms of the eyes. Individual cases of cholangiocarcinoma, hemangiosarcoma, and chemodectoma in dogs, as well as mammary gland cribriform carcinoma, salivary gland carcinoma, and histiocytic sarcoma in cats were detected.

Conclusions: The eyes of dogs were mostly affected by lymphoma or mammary gland carcinoma, while those of cats mostly had lymphoma, pulmonary carcinoma, or squamous cell carcinoma as the main metastatic/multicentric neoplasm. To the best of our knowledge, this is the first study to identify intraocular metastatic cholangiocarcinoma and chemodectoma in dogs, aside from identifying salivary gland carcinoma in cats.
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http://dx.doi.org/10.1111/vop.12803DOI Listing
September 2020

Atypical parasitic lesions in slaughtered cattle in Southern Brazil.

Rev Bras Parasitol Vet 2020 10;29(3):e001720. Epub 2020 Jul 10.

Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil.

In this study, we described the morphological features and unusual presentations of hydatidosis, fasciolosis, and eosinophilic myositis caused by Sarcocystis species diagnosed at the slaughterhouse lines. Twenty-seven samples of atypical parasitic lesions from distinct cattle were evaluated, of which 12 corresponded to hydatidosis, 11 to fasciolosis, and 4 to eosinophilic sarcocystosis. Hydatid cysts were observed mainly in the heart, with all cases involving the left ventricle. Fasciolosis lesions involved mainly the lungs, and were characterized by a focal nodular elevated area involving the edges of the lobes. Intralesional trematodes were observed in three cases. Sarcocystosis lesions were observed in four cases, and lesions were primarily located in the skeletal and cardiac muscles. Grossly, they presented as focal or focally extensive streaks, patches, or numerous nodules that were greenish to mildly yellowish. Histologically, all the cases had intralesional ruptured cysts of Sarcocystis that were associated with severe eosinophilic myositis and myocarditis. Parasitic lesions in atypical locations and/or with aberrant responses should be promptly identified because it may mistakenly diagnosed as other potentially zoonotic diseases, such as cysticercosis and tuberculosis.
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http://dx.doi.org/10.1590/s1984-29612020043DOI Listing
August 2020

Bovine abortion, stillbirth and neonatal death associated with Babesia bovis and Anaplasma sp. infections in southern Brazil.

Ticks Tick Borne Dis 2020 07 15;11(4):101443. Epub 2020 Apr 15.

Veterinary Pathology Department, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brazil.

Anaplasmosis and babesiosis are tick-borne diseases widely disseminated in cattle herds in many parts of the world. These diseases represent important causes of death and economic losses in several countries, including Brazil, and are characterized by hemolytic disease and anemia. Animals of all ages may be affected. Although transplacental infections are known to occur, abortion, stillbirth and neonatal death directly associated with Anaplasma marginale and especially Babesia spp. infections have rarely been documented in cattle. The objective of the present study is to describe the pathological and molecular findings of two cases of bovine abortion, two cases of stillbirth and two cases of neonatal death associated with intrauterine anaplasmosis and/or babesiosis in southern Brazil. All cases occurred in beef farms in the state of Rio Grande do Sul, between 2017 and 2019. Angus and crossbred calves were affected. At the necropsy, the main gross lesions observed included different degrees of splenomegaly, enlarged and yellow liver, thick and grumous bile, pallor or jaundice of mucous membranes and carcass, and dark kidneys. Four calves also presented cherry-pink discoloration of the central nervous system. Cytological slides enabled the observation of intraerythrocytic organisms consistent with Babesia bovis (3/6) and A. marginale (2/6). Through PCR assays, it was possible to detect three cases of Babesia sp. infection alone, and one case of Anaplasma sp. infection alone. Co-infections with Anaplasma sp. and Babesia sp. were detected in two cases. These findings reaffirm that anaplasmosis and babesiosis should be considered as an important differential diagnosis of fetal loss, stillbirth and neonatal death in cattle in areas where these diseases occur.
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http://dx.doi.org/10.1016/j.ttbdis.2020.101443DOI Listing
July 2020

Epidemiological, pathological and immunohistochemical aspects of 125 cases of feline lymphoma in Southern Brazil.

Vet Comp Oncol 2020 Jun 31;18(2):224-230. Epub 2019 Oct 31.

Setor de Patologia Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.

A retrospective study compiling cases of feline lymphoma diagnosed during 12 years (2004-2016) in Southern Brazil was performed. A total of 125 cases of lymphoma diagnosed in cats were reviewed, and information including age, breed, sex and tumour topography were collected. FeLV and FIV immunohistochemical tests were performed, as well as immunophenotyping of lymphomas. The alimentary form represented the most common presentation (42/125), followed by mediastinal lymphoma (35/125). Out of 125 cases, 79 presented positive retroviral immunostaining in tumour tissue (52 FeLV alone, 14 FIV alone and 13 presented FIV and FeLV co-infections), 66/125 of the cases were of T-cell origin and 59/125 of the cases were of B-cell origin. The median age of cats with T-cell lymphoma was 120 months (10-240 months), and 60 months (6-204 months) for cats with B-cell lymphoma. The most frequent alimentary tumour presentation was the enteropathy-associated T-cell lymphoma (type 1), and the major type of mediastinal tumour observed was diffuse large B-cell lymphoma. Considering only mediastinal and alimentary lymphomas (n = 77), the prevalence of mediastinal lymphoma in FeLV-positive cats was 2.21 times higher than the prevalence of this type of tumour in FeLV-negative cats (P = .036). Mediastinal lymphoma was more frequently observed in younger cats, and the prevalence of mediastinal tumours in these animals was 3.06 times higher than the prevalence of this tumour form in old cats (P = .0125). The present study indicates that retroviral infections still play an important role in the development of feline lymphomas in southern Brazil.
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http://dx.doi.org/10.1111/vco.12535DOI Listing
June 2020

Fatal parasite-induced enteritis and typhlocolitis in horses in Southern Brazil.

Rev Bras Parasitol Vet 2019 Aug 1;28(3):443-450. Epub 2019 Aug 1.

Setor de Patologia Veterinária, Departamento de Patologia Clínica Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul - UFRGS, Porto Alegre, RS, Brasil.

Diseases related to the alimentary system are the main cause of death in horses. This retrospective study aimed to describe the pathological findings of fatal parasite-induced enteritis and typhlocolitis caused by cyathostominae, Eimeria leuckarti, Balantidium coli, and Strongyloides westeri in horses. The records of parasite-induced intestinal lesions in horses necropsied in Southern Brazil between 2005 and 2017 were reviewed. Ten horses had fatal parasitic enteritis and/or typhlocolitis, and the main causes were: cyathostominae typhlocolitis (6/10), E. leuckarti enteritis (1/10), S. westeri enteritis (1/10), B. coli colitis related to cyathostominae (1/10), and infection by multiple agents (1/10). Cyathostominae typhlocolitis showed marked mucosal thickening, with multifocal elevated nodules containing tangled filiform parasites. Microscopic examination revealed that the mucosa and submucosa had encysted parasitic structures surrounded by eosinophilic and granulomatous inflammation. E. leuckarti enteritis was microscopically characterized by macrogamonts, microgamonts, and oocysts inside the host cells. S. westeri enteritis showed microscopic atrophy of the villi with numerous mucosal encysted parasitic structures. B. coli typhlocolitis showed severe diffuse mucosal reddening, with microscopic superficial mucosal necrosis associated with multiple protozoan trophozoites. Fatal parasite-induced enteritis and typhlocolitis are important causes of death in horses in Southern Brazil.
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http://dx.doi.org/10.1590/S1984-29612019056DOI Listing
August 2019

Meningoencephalitis due to Naegleria fowleri in cattle in southern Brazil.

Rev Bras Parasitol Vet 2019 Jun 6;28(3):514-517. Epub 2019 Jun 6.

Setor de Patologia Veterinária, Departamento de Patologia Clínica Veterinária, Universidade Federal do Rio Grande do Sul - UFRGS, Porto Alegre, RS, Brasil.

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.
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http://dx.doi.org/10.1590/S1984-29612019021DOI Listing
June 2019

Hydrallantois in cows naturally poisoned by in Brazil.

J Vet Diagn Invest 2019 Jul 23;31(4):581-584. Epub 2019 May 23.

Department of Veterinary Pathology, School of Veterinary Medicine, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil.

is a small subshrub that is distributed throughout Brazil and is responsible for lysosomal storage disease and occasional reproductive problems in cattle, goats, equids, sheep, and deer. We describe herein the clinical, epidemiologic, and pathologic features of hydrallantois in 3 cows naturally poisoned by in Rio Grande do Sul State, Brazil. Clinically, all cows had marked abdominal distension and mild ataxia. After natural death or euthanasia, autopsies revealed that the abdominal distension in all 3 cases was caused by severe enlargement of the uterus, which contained 100-120 L of translucent fluid within the allantois, in addition to adventitial placentation. Microscopic evaluation of the placenta revealed marked diffuse edema, sometimes with a myxomatous appearance. Neurons in the cerebellum and obex were swollen, with mild-to-moderate cytoplasmic granular vacuolation. Histochemical examination with lectins ConA, WGA, and sWGA revealed mild-to-marked staining in the cytoplasm of neurons of the cerebellum and medulla at the level of the obex, indicating the occurrence of α-mannosidosis.
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http://dx.doi.org/10.1177/1040638719850610DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6857035PMC
July 2019

Detection of enzootic nasal tumor virus (ENTV) in a sheep flock in southern Brazil.

Trop Anim Health Prod 2019 Sep 17;51(7):2095-2098. Epub 2019 Apr 17.

Department of Veterinary Pathology, Veterinary School, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.

Enzootic nasal tumor (ENT) is a contagious neoplasm associated with enzootic nasal tumor virus (ENTV), which may induce disease in sheep (ENTV-1) and goats (ENTV-2). This study aimed to describe the occurrence of ENT in two Texel sheep (Ovis aries) from a 75-sheep flock, located in the city of Gravataí, southern Brazil. Animals used to be purchased from different origins, and no specific tests for disease monitoring or quarantine procedure were performed. Affected animals presented respiratory distress, anorexia with severe weight loss, and mucopurulent unilateral nasal discharge. Necropsy was performed in both animals and nasal cavity masses were observed. Histopathological analysis demonstrated an epithelial neoplasm compatible with nasal adenocarcinoma. PCR using a protocol that amplifies a 591 bp sequence of 5'LTR-gag region of ENTV1 was performed followed by DNA sequencing. Both samples were positive, and the sequences obtained presented highest identity (97%) with ENTV strain TN28 (GenBank accession number MH899613) detected in a Texel sheep from Scotland. This is the first report of ENTV-1 leading to enzootic nasal tumor in sheep in Latin America, which confirms the presence of the retrovirus in sheep flocks in the Brazilian territory.
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http://dx.doi.org/10.1007/s11250-019-01897-zDOI Listing
September 2019

Presentation of hemolytic and hemorrhagic rangeliosis in Cerdocyon thous.

Ticks Tick Borne Dis 2019 04 22;10(3):690-693. Epub 2019 Feb 22.

Laboratório de Diagnóstico em Medicina Veterinária, Universidade de Caxias do Sul, R. Francisco Getúlio Vargas, 1130, Petrópolis, Caxias do Sul, RS, 95070560, Brazil. Electronic address:

Rangeliosis, caused by protozoan Rangelia vitalii, is transmitted by the tick Amblyomma aureolatum. The disease is characterized by hemolytic and hemorrhagic disorder and has been described in dogs and other wild canids. The aim of this study was to compare clinicopathological findings and laboratory results of a Rangelia infection in a crab-eating fox (Cerdocyon thous) with those of canine rangeliosis. The zoo of Universidade de Caxias do Sul, received a crab-eating fox with marked jaundice in mucous membranes, dark-colored stools and neurological signs. The animal underwent an ear tip smear examination and blood collection for complete blood counts, serum biochemistry and PCR. Free-living and intraerythrocytic pyriform structures consistent with R. vitalii were found in the blood smear of the ear tip. The erythrogram revealed normocytic normochromic anemia, moderate macrocytosis, polychromasia and metarubricytosis. The leukogram revealed leukocytosis with neutrophilia and monocytosis, as well as severe thrombocytopenia. Serum biochemistry showed hypoproteinemia, hypoalbuminemia and elevated levels of urea and creatinine. The treatment was performed with imidocarb hydrochloride and dexamethasone, however 24 h after initiation of treatment the animal died. Macroscopic examination revealed jaundice, subcutaneous edema, enlarged superficial lymph nodes, splenomegaly, and hemorrhage of internal organs. Histological sections of the cerebellum, lung, pancreas, intestine and heart were consistent with R. vitalii infection of the vascular endothelium. Pathological and hematological findings were similar to those found in infected dogs, with clinical presentation characterized by hemolytic anemia and hemorrhage. The description of this case showed that C. thous does not only serve as reservoir of R. vitalii but may also develop disease.
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http://dx.doi.org/10.1016/j.ttbdis.2019.02.010DOI Listing
April 2019

Omphalitis, urachocystitis and septicemia by Streptococcus dysgalactiae in a southern right whale calf Eubalaena australis, Brazil.

Dis Aquat Organ 2018 Nov;131(3):227-232

Setor de Patologia Veterinária, Faculdade de Veterinária (FAVET), Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul (RS) 91540-000, Brazil.

Southern right whales Eubalaena australis (SRW) use the southern coast of Brazil as a wintering and calving ground. Other than anthropogenic threats, there is limited knowledge on health and disease aspects for this species. We report the gross and microscopic findings and microbiological identification of streptococcal septicemia in a SRW calf. Main gross findings included fibrinosuppurative omphalitis and urachocystitis, suppurative cystitis, valvular endocarditis and myocarditis, embolic pneumonia, suppurative myositis and osteoarthritis, and lymphadenomegaly. Histological examination confirmed the above inflammatory processes and indicated disseminated Gram-positive coccoid septicemia. PCR analysis, based on the 16S rRNA gene from bacteria isolated on blood agar, identified Streptococcus dysgalactiae. Pathologic and microbiologic analysis indicated that β-hemolytic S. dysgalactiae septicemia, presumably initiated as ascending omphalic infection, was responsible for stranding and death in this individual. These results further confirm pathogenicity of streptococci in cetaceans and add to the limited health and disease related pathology knowledge for this species.
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http://dx.doi.org/10.3354/dao03293DOI Listing
November 2018

Angiostrongylosis in Cerdocyon thous (crab-eating fox) and Lycalopex gymnocercus (Pampas fox) in Southern Brazil.

Parasitology 2019 04 5;146(5):617-624. Epub 2018 Nov 5.

Department of Clinical Veterinary Pathology,Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul (UFRGS),Av. Bento Gonçalves 9090,42505, Porto Alegre, RS 91540-000,Brazil.

The objective of this study was to identify species of Angiostrongylus spp. infecting wild carnivores in Southern Brazil, as well as to describe gross and histopathological findings associated with the infection. Necropsy was conducted in 16 wild carnivores parasitized by Angiostrongylus spp. Analysed lungs revealed multifocal dark-red areas of consolidation; in one case, multifocal firm white nodules spread in all pulmonary lobes were observed. In one animal, a focally extensive area of malacia associated with haemorrhage was noted in the encephalon. Histologically, multifocal granulomatous pneumonia or bronchopneumonia, associated with eggs and larvae in blood vessels, lung interstitium, alveoli, and sometimes in bronchi and bronchioles was observed. Adult nematodes were seen within blood vessels. The lesion observed in the brain was characterized as a focally extensive area of malacia associated with gitter cells, haemorrhage, thrombosis and a free intralesional larva. Through molecular techniques, seven positive samples of Angiostrongylus cantonensis were obtained, including the brain sample, and a positive sample of Angiostrongylus vasorum-like, all in Cerdocyon thous. The positive sample for A. vasorum showed 97% similarity with sequences deposited in GenBank, suggesting a new species or subspecies of Angiostrongylus sp. Infection of Lycalopex gymnocercus by Angiostrongylus spp. was confirmed by histological evaluation.
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http://dx.doi.org/10.1017/S0031182018001865DOI Listing
April 2019

Mamastrovirus 5 detected in a crab-eating fox (Cerdocyon thous): Expanding wildlife host range of astroviruses.

Comp Immunol Microbiol Infect Dis 2018 Jun 15;58:36-43. Epub 2018 Aug 15.

Laboratório de Virologia, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9090, Prédio 42.602, CEP 91540-000, Porto Alegre, Rio Grande do Sul, Brazil. Electronic address:

Astroviruses are a common cause of gastroenteritis in children worldwide and can also cause infection in a range of domestic and wild animal species. Canine astrovirus (formally named as Mamastrovirus 5, MAstV5) has been reported worldwide, and its role as an enteric pathogen is still controversial. Herein, we describe the genomic characterization of a MAstV5 (strain crab-eating fox/2016/BRA) identified in a wild canid (Cerdocyon thous) diagnosed with canine distemper virus (CDV) as causa mortis. The nearly complete genome comprised 6579 nt in length and displayed the archetypal organization of astroviruses. The present report is the first evidence of MAstV5 infection in an animal species other than the dog and highlights a possible natural astrovirus spillover between domestic and wild canids. Moreover, these results show the first evidence of extra-intestinal MAstV5, suggesting a virus systemic spread. This work is expected to contribute to a better understanding of the astroviruses biology and their interactions with the wildlife health.
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http://dx.doi.org/10.1016/j.cimid.2018.08.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7112573PMC
June 2018

GASTRIC PYTHIOSIS IN A BACTRIAN CAMEL ( BACTRIANUS CAMELUS).

J Zoo Wildl Med 2018 Sep;49(3):784-787

Pythium insidiosum, an aquatic oomycete, causes chronic lesions in the skin and digestive tract of multiple species. A captive-bred Bactrian camel ( Camelus bactrianus) showed clinical signs of lethargy and weight loss in a clinical course of 30 days, with no response to treatment. At necropsy, the abdominal cavity had approximately 32 L of a yellow, turbid fluid with fibrin. The third compartment of the stomach (C-3) showed a focal area of rupture covered with fibrin. Close to this area, the C-3 wall was thickened and firm, demonstrating irregular, yellow, and friable areas on cut surface (kunkers). Microscopically, these corresponded to necrosis, characterized by a central amorphous eosinophilic material, surrounded by a pyogranulomatous inflammatory infiltrate and fibrosis. Negatively stained hyphae were observed at the periphery of the necrotic areas, which showed marked immunostaining for P. insidiosum. Pythiosis in camelids may involve the stomach, resulting in peritonitis and death.
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http://dx.doi.org/10.1638/2017-0195.1DOI Listing
September 2018

Canine distemper virus active infection in order Pilosa, family Myrmecophagidae, species Tamandua tetradactyla.

Vet Microbiol 2018 Jul 25;220:7-11. Epub 2018 Apr 25.

Laboratory of Animal Virology, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Celso Garcia Cid Road, PR 445 Km 380, 86051-990, Londrina, PR, Brazil.

Canine distemper virus (CDV) is a highly contagious disease pathogen which causes disease in the domestic dog and species classified in the Canidae, Procyonidae, Mustelidae, Hyaenidae, Ursidae, Viveridae, Felidae, Tayassuidae, and Cercopithecidae families. A combined strategy that involved the direct sequencing of amplicons from genes coding for nucleocapsid, large polymerase, and hemagglutinin proteins of CDV, as well as the pathological findings and the immunohistochemical detection of viral nucleocapsid protein in diverse tissues, confirmed the participation of CDV in the development of a neurological disease in a southern tamandua (Tamandua tetradactyla) from Midwestern Brazil. Phylogenetic analysis based on the hemagglutinin gene sequences revealed that the strain from this study grouped with isolates from the Europe 1/South America 1 lineage. The specific polymorphisms at the SLAM receptor-binding site of the hemagglutinin gene, previously linked to disease emergence in novel hosts, were not detected in this genome. These findings represent the first description of CDV-induced infection in the Tamandua tetradactyla and extend the distribution of this infection to include members of the family Myrmecophagidae, order Pilosa.
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http://dx.doi.org/10.1016/j.vetmic.2018.04.030DOI Listing
July 2018

Identification of bluetongue virus serotypes 1, 4, and 17 co-infections in sheep flocks during outbreaks in Brazil.

Res Vet Sci 2017 Aug 5;113:87-93. Epub 2017 Sep 5.

Setor de Patologia Veterinária - Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9090, Porto Alegre CEP 91540-000, Brazil.

Bluetongue (BT) is a vector-borne viral disease caused by the Bluetongue virus (BTV), an Orbivirus from the Reoviridae family, affecting domestic and wild ruminants. BTV circulation in Brazil was first reported in 1978, and several serological surveys indicate that the virus is widespread, although with varied prevalence. In 2014, BT outbreaks affected sheep flocks in Rio Grande do Sul state, causing significant mortality (18.4%; 91/495) in BTV-infected sheep. In total, seven farms were monitored, and one or two sheep from each farm that died due to clinical signs of BT were necropsied. Apathy, pyrexia, anorexia, tachycardia, respiratory, and digestive disorders were noted. Additionally, an abortion was recorded in one of the monitored farms. The main gross lesions observed were pulmonary edema, anterior-ventral pulmonary consolidation, muscular necrosis in the esophagus and in the ventral serratus muscle, and hemorrhagic lesions in the heart. The blood and tissue samples were tested for BTV RNA detection by RT-qPCR targeting the segment 10. Positive samples were used for viral isolation. The isolated BTVs were typed by conventional RT-PCR targeting the segment 2 of the 26 BTV serotypes, followed by sequencing analysis. BTV-1, BTV-4 and BTV-17 were identified in the analyzed samples. Double or triple BTV co-infections with these serotypes were detected. We report the occurrence of BT outbreaks related to BTV-1, BTV-4 and BTV-17 infections and co-infections causing clinical signs in sheep flocks in Southern Brazil, with significant mortality and lethality rates.
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http://dx.doi.org/10.1016/j.rvsc.2017.09.001DOI Listing
August 2017

Osteitis fibrosa cystica in a domestic young cat.

JFMS Open Rep 2015 Jul-Dec;1(2):2055116915607555. Epub 2015 Oct 12.

Sector of Veterinary Pathology, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.

Case Summary: A 4-month-old cat had bilateral swellings of the mandible, maxilla, humerus and femur, and angular deviations in the axial and appendicular skeleton. The biochemical profile indicated hypercalcemia, hyperphosphatemia and increased parathyroid hormone levels. Because of the poor prognosis, the cat was euthanized. At necropsy, malleable and fragile bones, associated with numerous cystic areas containing yellowish and translucent liquid, were observed. Histologically, the bones showed marked diffuse proliferation of fibrous connective tissue, and large numbers of osteoclasts surrounding numerous cystic structures were also observed within fibrotic areas at the periphery of the trabecular bone. In addition, enlargement of the parathyroid glands, which was associated with increased serum concentrations of calcium, phosphorus and parathyroid hormone, was detected.

Relevance And Novel Information: The changes observed in this cat are consistent with hyperparathyroidism-associated osteitis fibrosa cystica, which is an unusual presentation in the cat. Hyperparathyroidism, either primary (neoplastic) or secondary (nutritional or renal), is the primary cause of this condition.
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http://dx.doi.org/10.1177/2055116915607555DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5361987PMC
October 2015

Natural Infection of Wild Canids (Cerdocyon thous and Lycalopex gymnocercus) with the Intraendothelial Piroplasm Rangelia vitalii in Southern Brazil.

J Wildl Dis 2015 Oct 7;51(4):880-4. Epub 2015 Aug 7.

1  Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, CEP 91540-000, Porto Alegre, Rio Grande do Sul, Brazil.

Rangelia vitalii is a piroplasm that infects canines, causing lesions typical of a hemolytic disorder. Two wild canids, a crab-eating fox (Cerdocyon thous) and a Pampas fox (Lycalopex gymnocercus), were presented for necropsy in Setor de Patologia Veterinária at the Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil. On gross examination, both animals had pale mucosae and moderate tick infestation (Amblyomma aureolatum). There was severe splenomegaly, and the liver had a diffusely orange-reddish lobular pattern. The mesenteric lymph nodes were brownish and slightly enlarged. Structures compatible with R. vitalii were observed in the cytoplasm of endothelial cells in the liver, stomach, heart, kidney, lungs, lymph nodes, and bladder. The agent was characterized by PCR and genetic sequencing of liver samples and ticks. We show that parasitism with R. vitalii follows an epidemiologic cycle in which wild canids act as reservoirs.
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http://dx.doi.org/10.7589/2014-12-283DOI Listing
October 2015
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