Abstract in English:
Leishmaniasis is a zoonotic disease caused by parasites of the Leishmania genus, resulting in various clinical forms, including a highly lethal visceral form. This study aimed to identify Leishmania spp. in sandflies and dogs in a small Brazilian town. DPP® Visceral Canine Leishmaniasis (CVL) was used for screening, and ELISA and conventional PCR were used for confirmatory testing, while sandflies were captured using CDC light traps and conventional PCR targeting ITS1. The Hill series was used to identify the diversity profile of species in the sampled area using R software. The study identified a Leishmania spp. prevalence of 4.02% in dogs, exceeding the 2% limit required by the Brazilian Ministry of Health. A total of 443 sandflies belonging to 14 different species were identified, with Lutzomyia longipalpis being the most abundant (73.81%). Negligence regarding leishmaniasis in small towns can lead to late diagnosis, hence the need to implement effective strategies, including early diagnosis and treatment of human and canine cases, vector control programs, and awareness campaigns to educate the public about risks and preventive measures. These measures can help prevent the spread of leishmaniasis and improve health outcomes for affected individuals and animals.
Abstract in Portuguese:
A leishmaniose é uma doença zoonótica causada por parasitas do gênero Leishmania, resultando em várias formas clínicas, incluindo uma forma visceral altamente letal. Este estudo teve como objetivo identificar Leishmania spp. DNA em flebotomíneos e cães em uma pequena cidade brasileira. A investigação usou DPP® Visceral Canine Leishmaniasis (CVL) para triagem, e ELISA e PCR convencional para teste confirmatório, enquanto flebotomíneos foram capturados usando armadilhas de luz CDC e PCR convencional visando ITS1. A série Hill foi utilizada para identificar o perfil de diversidade das espécies na área amostrada, utilizando o software R. O estudo identificou uma prevalência de Leishmania spp. em 4,02% dos cães, ultrapassando o limite de 2% exigido pelo Ministério da Saúde do Brasil. Foi identificado um total de 443 flebotomíneos, pertencentes a 14 espécies diferentes, com Lutzomyia longipalpis sendo o mais abundante (73,81%). A negligência com a leishmaniose em cidades pequenas pode levar ao diagnóstico tardio, daí a necessidade de implementar estratégias eficazes, incluindo diagnóstico precoce e tratamento de casos humanos e caninos, programas de controle de vetores e campanhas de conscientização para educar o público sobre riscos e medidas preventivas. Essas medidas podem ajudar a prevenir a propagação da leishmaniose e melhorar os resultados de saúde para indivíduos e animais afetados.
Abstract in English:
Ophidian accidents have been a medical and veterinary challenge. The only treatment available and recommended by the World Health Organization and the Ministry of Health is serotherapy. This study evaluated the liver function of equines submitted to the immunization protocol currently used by the “Instituto Vital Brazil” to produce anti-Bothrops serum. Five equines were evaluated with Bothrops venom inoculation during three immunization cycles. Blood samples were taken for a biochemical test and a liver assessment due to the damage caused to the liver. The biochemical data that presented significant differences in all animals compared to baseline were AST, ALT, GGT, FA, GLDH and serum protein. Creatinine did not change throughout the experiment. The results obtained suggest that liver alterations may occur throughout the three immunization cycles. There is a great need for new studies aimed at broadening knowledge about venom inoculation protocols in serum producer equines and minimizing the adverse effects of immunization.
Abstract in Portuguese:
Acidentes ofídicos têm sido um desafio médico e veterinário. O único tratamento disponível e preconizado pela Organização Mundial da Saúde e pelo Ministério Saúde é a soroterapia. O presente estudo avaliou a função hepática dos equinos submetidos ao protocolo de imunização empregado atualmente pelo Instituto Vital Brazil para produção do soro antibotrópico. Foram avaliados cinco equinos com inoculação de veneno Botrópico durante o período de três ciclos de imunizações. Amostras de sangue foram coletadas para realização de exame bioquímico e avaliação hepática por conta das lesões causadas no fígado. Os dados bioquímicos que apresentaram diferenças significativas, em todos os animais, comparados ao tempo basal, foram AST, ALT, GGT, FA, GLDH e proteína sérica. Creatinina não apresentou alteração ao longo do experimento. Os resultados obtidos sugerem que ao longo dos três ciclos de imunização podem ocorrer alterações hepáticas. Há uma grande necessidade de novos estudos com objetivo de ampliar o conhecimento acerca dos protocolos de inoculação de veneno nos equinos soroprodutores e minimizar os efeitos adversos da imunização.
Abstract in English:
Crotalaria spectabilis seed poisoning has been reported in cattle on rural properties in Diamantino and Campo Verde municipalities in Mato Grosso, Brazil. The disease appeared 75 to 120 days after the animals were fed a diet contaminated with C. spectabilis seeds. Clinically, the animals showed a lack of appetite, progressive weight loss, apathy, photosensitivity, lateral recumbency, and death. Some animals developed neurological manifestations as a consequence of hepatic encephalopathy. In total, 37 cattle died, and six cattle were necropsied. The main macroscopic changes consisted of ascites and a firm and decreased liver with an irregular surface and multifocal pale yellowish discoloration. Microscopically, hepatocyte loss with fibrosis, megalocytosis and bile duct proliferation was observed. Animals with severe hepatic damage also had status spongiosus in the central nervous system. Furthermore, a hepatic biopsy of 15 cows who consumed contaminated grains but did not develop clinical disease revealed histologic liver changes similar to necropsied animals.
Abstract in Portuguese:
Relata-se dois surtos de intoxicação por sementes de Crotalaria spectabilis em bovinos em propriedades rurais nos municípios de Diamantino e Campo Verde, Mato Grosso, Brasil. A doença apareceu 75 a 120 dias depois que os animais foram alimentados com uma dieta contaminada com sementes de C. spectabilis. Clinicamente, foi observado falta de apetite, perda de peso progressiva, apatia, fotossensibilização, decúbito lateral e morte. Alguns animais desenvolveram manifestações neurológicas secundárias à encefalopatia hepática. No total, 37 bovinos morreram e seis deles foram submetidos à necropsia. As principais alterações macroscópicas consistiram em ascite e fígado diminuído em tamanho com superfície capsular irregular, difusamente firme e com áreas multifocais amarelo-pálidas. Microscopicamente, foi observado perda de hepatócitos acompanhada de fibrose hepática com megalocitose e proliferação de ductos biliares. No sistema nervoso central de bovinos com lesões hepáticas graves havia status spongiosus. A biópsia hepática de 15 bovinos que consumiram grãos contaminados, mas estavam clinicamente sadios, revelou alterações hepáticas similares às descritas nos animais submetidos à eutanásia.
Abstract in English:
The state of Santa Catarina is the largest national producer of onions. The consumption of onion (Allium cepa) by domestic animals can cause hemolytic anemia and hyperhemoglobinemia of Heinz bodies in erythrocytes. The epidemiological data and the pathological clinical picture were obtained during a visit to the property where the disease occurred. A herd of 54 beef cattle from a property in the municipality of Bom Retiro/SC, was placed in an onion crop, which had not been completely harvested because it was outside the desired standard for trade. The animals remained in this field for eight days. At the end of this period, all of them presented apathy, anorexia, brown urine and mucous membranes and an onion odor on expiration. The macroscopic changes observed were pale mucous membranes, blood with decreased viscosity and increased clotting time, liver with an evident lobular pattern and remains of onion in the rumen. The carcass gave off a strong onion odor. By histological analysis, there was central lobular coagulation necrosis in bridging in the liver. In the kidneys, there were intracytoplasmic hyaline droplets in tubular epithelial cells and a deposit of eosinophilic material in the lumen of the tubules. For the diagnosis of onion poisoning, epidemiological data associated with clinical signs, such as pale mucous membranes with evidence of hemoglobin pigmentation (brown mucosa) and hemoglobinuria, should be taken into account. This article seems to be the first report of onion poisoning in cattle in Brazil.
Abstract in Portuguese:
O estado de Santa Catarina é o maior produtor nacional de cebola. O consumo de cebola (Allium cepa) por animais domésticos pode causar anemia hemolítica, hiper-hemoglobinemia de corpúsculos de Heinz nos eritrócitos. Os dados epidemiológicos e o quadro clínico-patológico foram obtidos em visita à propriedade onde a enfermidade ocorreu. Um rebanho de 54 bovinos de corte de uma propriedade localizada no município de Bom Retiro/SC, foi colocado em uma lavoura de cebola, a qual não havia sido completamente colhida por estar fora do padrão desejado para o comércio. Os animais permaneceram por um período de oito dias nesta lavoura e, ao final deste período, todos apresentaram apatia, anorexia, urina e mucosas de coloração marrom e odor de cebola na expiração. As alterações macroscópicas observadas foram mucosas pálidas, sangue com viscosidade diminuída e aumento do tempo de coagulação, fígado com padrão lobular evidente e restos de cebola no rúmen. A carcaça exalava forte odor de cebola. Pela análise histológica havia, no fígado, necrose de coagulação centrolobular em ponte e nos rins havia gotas hialinas intracitoplasmáticas em células do epitélio tubular e depósito de material eosinofílico na luz de túbulos. Para o diagnóstico da intoxicação por cebola deve-se levar em conta principalmente os dados epidemiológicos associados aos sinais clínicos, como palidez de mucosas com evidências de pigmentação por hemoglobina (mucosa marrom) e hemoglobinúria. Este artigo parece ser o primeiro relato de intoxicação por cebola em bovinos no Brasil.
Abstract in English:
Feline injection-site sarcomas in felines account for more than 40% of cutaneous and subcutaneous neoplasms in felines. The present study aimed to describe the macroscopic and histomorphological findings of feline application/injection sarcomas. Samples from 31 feline tumors with a history of feline application/inflammation sarcoma were re-evaluated regarding histological subtype, mitotic index and score, depth of tissue invasion, and presence of inflammation considering the location, intensity and predominant cell types. Of the 31 samples from felines diagnosed with sarcoma at the application/inflammation site, 87.15% were cats with no defined breed (NDB), with a mean age of 8.5 years. The predominant anatomical sites were the back and flank/abdomen, both with 29% (9/31), and the prevalent histological subtype was fibrosarcoma at 77.4% (24/31), followed by anaplastic giant cell sarcoma at 12.9% (4/31) and myxosarcoma 9.6% (3/31). The histological grade with the highest number of cases was III (51.6%), followed by Grade II (35.4%) and I (12.9%). The mean of the longest axis measurements varied between the different tumor grades without being significant, with the average being 2.5±2.79cm in Grade I tumors and 3.2±2.28cm in Grade II tumors. and 4.68±2.07cm in Grade III tumors. Necrosis was observed in 74.2% of tumors. The tissue inflammation score was mild to moderate in 58% of cases and severe in 32.2%, with lymphocytic and lymphoplasmacytic infiltrates being prevalent, with 25.8% each, followed by lymphoplasmohistiocytic with 22.6%. The infiltration depth was 38.7% in muscle tissue, followed by 32.2% in the subcutaneous tissue. Pleomorphism was accentuated in 51.6%. Desmoplasia was moderate in 45.1%. Satellite nodules were present in 29% of cases, and 19.4% had macrophages with intracytoplasmic content suggestive of adjuvants. Surgical margins were infiltrated (M1) in 48.4% and narrowed in 25.8% (M2). The anatomical locations observed were different from those recommended by the Vaccine-Associated Feline Sarcoma Task Force (VAFST); in most cases, the adjuvanted macrophage was not present. From this data, we can suggest that sarcomas in felines are not only correlated to the vaccine application, corroborating the hypothesis that any material, whether liquid or solid, and any chronic inflammatory process in the subcutaneous tissue of cats can induce the entity if they are predisposed to do so. The detailed histomorphological data evaluated in this study were key points and provided important information about tumor behavior, being a tool for clinical-oncological decision-making.
Abstract in Portuguese:
Feline injection-site sarcomas in felines account for more than 40% of cutaneous and subcutaneous neoplasms in felines. The present study aimed to describe the macroscopic and histomorphological findings of feline application/injection sarcomas. Samples from 31 feline tumors with a history of feline application/inflammation sarcoma were re-evaluated regarding histological subtype, mitotic index and score, depth of tissue invasion, and presence of inflammation considering the location, intensity and predominant cell types. Of the 31 samples from felines diagnosed with sarcoma at the application/inflammation site, 87.15% were cats with no defined breed (NDB), with a mean age of 8.5 years. The predominant anatomical sites were the back and flank/abdomen, both with 29% (9/31), and the prevalent histological subtype was fibrosarcoma at 77.4% (24/31), followed by anaplastic giant cell sarcoma at 12.9% (4/31) and myxosarcoma 9.6% (3/31). The histological grade with the highest number of cases was III (51.6%), followed by Grade II (35.4%) and I (12.9%). The mean of the longest axis measurements varied between the different tumor grades without being significant, with the average being 2.5±2.79cm in Grade I tumors and 3.2±2.28cm in Grade II tumors. and 4.68±2.07cm in Grade III tumors. Necrosis was observed in 74.2% of tumors. The tissue inflammation score was mild to moderate in 58% of cases and severe in 32.2%, with lymphocytic and lymphoplasmacytic infiltrates being prevalent, with 25.8% each, followed by lymphoplasmohistiocytic with 22.6%. The infiltration depth was 38.7% in muscle tissue, followed by 32.2% in the subcutaneous tissue. Pleomorphism was accentuated in 51.6%. Desmoplasia was moderate in 45.1%. Satellite nodules were present in 29% of cases, and 19.4% had macrophages with intracytoplasmic content suggestive of adjuvants. Surgical margins were infiltrated (M1) in 48.4% and narrowed in 25.8% (M2). The anatomical locations observed were different from those recommended by the Vaccine-Associated Feline Sarcoma Task Force (VAFST); in most cases, the adjuvanted macrophage was not present. From this data, we can suggest that sarcomas in felines are not only correlated to the vaccine application, corroborating the hypothesis that any material, whether liquid or solid, and any chronic inflammatory process in the subcutaneous tissue of cats can induce the entity if they are predisposed to do so. The detailed histomorphological data evaluated in this study were key points and provided important information about tumor behavior, being a tool for clinical-oncological decision-making.
Abstract in English:
Local adverse reactions following vaccination, often tied to the adjuvant type, can lead to pain, fever, reduced intake, and weight loss. Infrared thermography (IRT), a non-invasive human and veterinary medicine technique, can assess local inflammation. This study aimed to evaluate such reactions induced by reproductive vaccines with different adjuvants, using IRT alongside cardinal signs, rectal temperature, and haptoglobin concentration. Thirty-five Holstein heifers were grouped by vaccine type: Ah (aluminum hydroxide), Ow (oil-in-water), QAD (amphigen and quil A cholesterol and dimethyl-dioctadecyl ammonium bromide adjuvant), and a Control (saline solution). Assessments were made at 0, 6, 24-, 48-, 72-, and 168 hours post-vaccination for both doses, with an interval of 21 days. The local reactions were evaluated using the inflammatory cardinal signs and surface temperature measurement using IRT. The systemic reactions were identified by rectal temperature and the concentration of haptoglobin. A larger proportion of animals exhibiting local reactions based on scores assigned to the cardinal signs was found in the Vaccine QAD group, as well as the rectal temperature and the frequency of heifers with inflammation (Hp ≥2mg/dL). Nevertheless, Vaccine Ow demonstrated higher temperature at the site after the first vaccination dose for the IRT. Therefore, this approach is a valuable tool in classifying responses and local inflammation following vaccination in heifers with reproductive vaccines. Concurrently evaluating systemic manifestations, facilitates the surveillance of adverse reactions, thereby improving the discernment of the extent of systemic and local effects.
Abstract in Portuguese:
Reações adversas locais após a vacinação, frequentemente associadas ao tipo de adjuvante, podem levar a dor, febre, redução na ingestão e perda de peso. A termografia infravermelha (IRT), uma técnica não invasiva utilizada na medicina humana e veterinária, oferece um meio de avaliar a inflamação local. Este estudo teve como objetivo avaliar tais reações induzidas por vacinas reprodutivas com diferentes adjuvantes, utilizando IRT junto com sinais cardinais, temperatura retal e concentração de haptoglobina. Trinta e cinco novilhas Holandesas foram agrupadas por tipo de vacina: Ah (hidróxido de alumínio), Ow (óleo em água), QAD (anfígeno e adjuvante de quil A colesterol e brometo de amônio dimetil-dioctadecil) e Controle (solução salina). As avaliações foram realizadas às 0, 6, 24, 48, 72 e 168 horas pós-vacinação para ambas as doses, com um intervalo de 21 dias. As reações locais foram avaliadas pelos sinais cardinais inflamatórios e pela aferição da temperatura superficial usando IRT. As reações sistêmicas foram identificadas pela temperatura retal e pela concentração de haptoglobina (Hp). Uma proporção maior de animais exibindo reações locais com base nos escores atribuídos aos sinais cardinais foi encontrada no grupo Vacina QAD, assim como na temperatura retal e na frequência de novilhas com inflamação (Hp ≥2mg/dL). No entanto, para a IRT, a Vacina Ow demonstrou temperatura mais elevada no local após a primeira dose de vacinação. Portanto, essa abordagem é uma ferramenta valiosa para classificar as respostas e a inflamação local após a vacinação em novilhas com vacinas reprodutivas. Avaliar simultaneamente as manifestações sistêmicas facilita a vigilância das reações adversas, melhorando assim o discernimento da extensão dos efeitos tanto sistêmicos quanto locais.
Abstract in English:
The Caatinga biome is unique to Brazil, with unfavorable environmental characteristics for the survival of Leptospira spp. However, recent studies have shown high positivity at PCR (polymerase chain reaction) in small ruminants. There are no Leptospira spp. studies based on sample calculation in pigs in the Caatinga. The aim of this study was to assess the importance of pigs in the spread of leptospirosis in the Caatinga biome. Overall, 200 biological samples (urine, blood, vaginal fluid, and tissues of reproductive and urinary tracts) were collected from 40 slaughtered sows, and MAT (microscopic agglutination test) and PCR tests were carried out to detect anti-Leptospira spp. antibodies and the agent’s DNA, respectively. The serological analysis showed a positivity rate of 5% (2/40), and the PCR identified Leptospira spp. DNA in 62.5% (25/40) of the animals. Only 2.5% (1/40) of the animals were positive for both techniques. The detected serogroups were Australis (50%) and Bataviae (50%), with antibody titers of 25 and 50. Leptospira spp. DNA was detected in 40% (16/40) of the reproductive tract samples, 32.5% (13/40) of the urinary tract, 32.5% (13/40) of the vaginal fluid and 30% (12/40) of the urine. There was no agreement (Kappa <0) between PCR samples from the genital tract vs. urinary tract or serological results. Genetic sequencing of one urine and one urinary tract tissue sample revealed 99% identity with L. borgpetersenii. The results indicate that leptospirosis is a concern in pigs in the context of Caatinga, with a high prevalence of infection detected by different diagnostic methods. The molecular analysis revealed a considerable proportion of infected animals. The findings emphasize the importance of a multifaceted approach in the diagnosis of leptospirosis in pigs, with a focus on the use of genital tract samples for the diagnosis of leptospirosis in this animal species, providing valuable insights for the control and prevention of this disease in both animals and the zoonotic context. Finally, the detection of leptospires in the genital tract indicates a possibility of male-female transmission in the venereal context.
Abstract in Portuguese:
O bioma Caatinga é único no Brasil, com características ambientais desfavoráveis à sobrevivência de Leptospira spp. Porém, estudos recentes demonstraram alta positividade na PCR (reação em cadeia da polimerase) em pequenos ruminantes. Não existem estudos para a infecção por Leptospira spp. baseados em cálculo amostral em suínos na Caatinga. O objetivo deste estudo foi avaliar a importância dos suínos na disseminação da leptospirose no bioma Caatinga. Foram coletadas 200 amostras biológicas (urina, sangue, fluido vaginal e tecidos do trato reprodutivo e urinário) de 40 porcas abatidas e realizados testes SAM (teste de soroaglutinação microscópica) e PCR para detecção de anticorpos anti-Leptospira spp. e DNA do agente, respectivamente. A análise sorológica mostrou taxa de positividade de 5% (2/40) e a PCR identificou o DNA de Leptospira spp. em 62,5% (25/40) dos animais. Apenas 2,5% (1/40) dos animais foram positivos para ambas as técnicas. Os sorogrupos detectados foram Australis (50%) e Bataviae (50%), com títulos de anticorpos de 25 e 50. O DNA de Leptospira spp. foi detectado em 40% (16/40) das amostras do trato reprodutivo, 32,5% (13/40) do trato urinário, 32,5% (13/40) do fluido vaginal e 30% (12/40) de urina. Não houve concordância (Kappa <0) entre amostras de PCR do trato genital vs. trato urinário ou resultados sorológicos. O sequenciamento genético de uma amostra de urina e de uma amostra de tecido do trato urinário revelou 99% de identidade com L. borgpetersenii. Os resultados obtidos indicam que a leptospirose representa uma preocupação em suínos no contexto da Caatinga, com alta prevalência de infecção detectada por diferentes métodos diagnósticos, bem como análises moleculares revelaram proporção considerável de animais infectados. Os resultados enfatizam a importância de uma abordagem multifacetada no diagnóstico da leptospirose em suínos, com foco no uso de amostras do trato genital para o diagnóstico da leptospirose nesta espécie animal, fornecendo informações valiosas para o controle e prevenção desta doença em animais e no contexto zoonótico. Por fim, a detecção de leptospiras no trato genital indica possibilidade de transmissão macho-fêmea no contexto venéreo.
Abstract in English:
Lymph node status is considered an important clinical prognostic factor in canine mammary carcinomas and women’s breast neoplasms. However, occult isolated tumor cells (ITCs) can be missed during hematoxylin and eosin (HE) analyses. Immunohistochemistry (IHC) for cytokeratin can be used to detect carcinomatous occult ITCs in mammary drainage lymph nodes. However, brown pigments, such as hemosiderin and ceroid in lymph nodes, may hinder the search for occult metastases by IHC utilizing DAB (3,3′-diaminobenzidine) as the chromogen. The aim of this study was to identify ITCs in canine lymph nodes of cases in which it was not detectable by routine HE evaluation through IHC for cytokeratin (AE1/AE3) combined with histochemistry techniques, such as Perls’ Prussian blue and periodic acid-Schiff (PAS), to improve the detection of occult metastases when hemosiderin and ceroid were present in these lymph nodes. For this, 25 tubulopapillary mammary carcinomas with their respective submitted 29 regional lymph nodes, previously given as free of tumor cells by HE analyses, were selected. Mammary tumors were graduated, and vascular invasion was investigated in these tumors. The submitted lymph nodes were reevaluated in HE, looking for occult metastases. IHC for cytokeratin (AE1/AE3) was used to detect occult metastases in mammary lymph nodes. Subsequently, a combined technique of IHC with Perl’s Prussian blue (for hemosiderin) or PAS (for ceroid) was performed to optimize the detection of ITCs by IHC, distinguishing them from pigments. Occult metastases were classified by their microanatomical location in subcapsular, cortical and medullary. Hemosiderin and ceroid were searched in lymph nodes and quantified as low, moderate, or high. The amount of pigments with a percentage of ITCs was also compared. Isolated tumor cells were found in 24.1% (7/29) of mammary lymph nodes. These ITCs were located mainly in subcapsular sinuses (4/7; 57.1%), followed by cortical (2/7; 28.5%) and medullary sinuses (1/7; 14.3%). There were concomitant lymph nodes with ITCs in 33.4% (2/6) of cases with vascular invasion. Hemosiderin and ceroid were present in about 90% of the 29 lymph nodes analyzed. In 42.8% (3/7) of lymph nodes with ITCs, hemosiderin and/or ceroid were in the same location as ITCs. It was found that lymph nodes in which ITCs were detected also present high amounts of hemosiderin (3/7; 42.9%) and low amounts of ceroid (5/7; 71.4%). In this study, IHC for cytokeratin (AE1/AE3) was an efficient method to detect occult tumor cells. IHC combined with Perls’ Prussian blue or with PAS proved to be a helpful way to investigate the presence of occult metastases in the lymph nodes of mammary canine tumors. It allowed distinguishing hemosiderin and ceroid, respectively, from ITCs in the same slide of IHC (immunostained by DAB), favoring a more accurate analysis by pathologists, which can be useful for the oncological staging of these patients.
Abstract in Portuguese:
O status nodal é considerado um importante fator prognóstico clínico em cães com carcinomas mamários e em mulheres com neoplasmas mamários. No entanto, durante a análise histopatológica convencional (coloração de hematoxilina e eosina - HE), células neoplásicas podem permanecer ocultas no linfonodo. Imuno-histoquímica (IHQ) para citoqueratina pode ser utilizada para detectar células carcinomatosas isoladas ocultas em linfonodos de drenagem mamária. Entretanto, a presença de pigmentos marrons, como a hemossiderina e o ceroide em linfonodos, pode dificultar a procura por metástases ocultas durante a análise por IHQ utilizando o DAB (3,3′-diaminobenzidina) como cromógeno. O objetivo deste estudo foi identificar células tumorais isoladas (CTIs), não detectadas pela avaliação de rotina (HE), em linfonodos de cães, através de IHQ para citoqueratina (AE1/AE3) combinada com técnicas histoquímicas, como azul da Prússia e ácido periódico Schiff (PAS), para otimizar a detecção de metástases ocultas, quando os pigmentos hemossiderina e ceroide estavam presentes nos linfonodos. Para esta finalidade, 25 carcinomas mamários túbulo-papilares com seus respectivos 29 linfonodos regionais remetidos, previamente diagnosticados como livres de células neoplásicas pelo HE foram selecionados. Os tumores mamários foram graduados e a presença de invasão vascular foi investigada nestes tumores. Os linfonodos submetidos foram reavaliados histologicamente no HE, à procura de metástases ocultas. IHQ para citoqueratina (AE1/A3) foi utilizada para detectar metástases ocultas nos linfonodos mamários. Subsequentemente, técnicas combinadas de IHQ com azul da Prússia (para hemossiderina) ou PAS (para ceroide) foram realizadas para otimizar a detecção de CTIs através da IHQ, distinguindo-as dos pigmentos. As metástases ocultas foram classificadas pela sua região microanatômica nodal em subcapsular, cortical e medular. Hemossiderina e ceroide foram identificados no linfonodo e classificados de acordo com sua quantidade em baixa, moderada e alta. Também foram comparadas a quantidade de pigmentos com a porcentagem de CTIs. Foram encontradas CTIs em 24,1% (7/29) dos linfonodos mamários. Estas CTIs estavam localizadas principalmente em seios subcapsulares (4/7; 57,1%), seguido pela cortical (2/7; 28,5%) e seios medulares (1/7; 14,3%). Em 33,4% (2/6) dos casos com invasão vascular havia, concomitantemente, linfonodos de drenagem com CTIs. Hemossiderina e ceroide estavam presentes em cerca de 90% dos 29 linfonodos analisados. Em 42,8% (3/7) dos linfonodos com CTIs, hemossiderina e/ou ceroide estavam na mesma localização das CTIs. Os linfonodos que apresentaram CTIs também apresentavam grande quantidade de hemossiderina (3/7; 42,9%) e pequena quantidade de ceroide (5/7; 71,4%). Neste estudo, IHQ para citoqueratina (AE1/AE3) foi um método eficiente para detecção de células tumorais ocultas. A técnica combinada de IHQ com azul da Prússia ou com o ácido periódico de Schiff (PAS) mostrou ser uma maneira útil de investigar a presença de metástases ocultas em linfonodos de cães com tumores mamários caninos, distinguindo a hemossiderina e o ceroide, respectivamente, de CTIs na mesma lâmina de IHQ (imunomarcadas por DAB), favorecendo uma análise mais acurada pelos patologistas, o que pode ser útil no estadiamento oncológico destes pacientes.
Abstract in English:
This study aimed to determine the frequency of anti-Neospora caninum antibody detection in three samples and sampling methods: A prevalence study, routine diagnostic laboratory samples, and fetal bovine serum samples. These samples were collected from cattle in Rio Grande do Sul (RS), southern Brazil, and analyzed using the indirect immunofluorescence reaction technique. For each sampling method, a historical study was used as a reference for comparison. In the prevalence study, 1,248 serum samples were collected from 2020 to 2022. The prevalence of N. caninum in the RS state was 22.8% (285/1248). This figure was statistically different compared to previous studies conducted in 2002, which reported a prevalence of 11.2% (p<0.001). In the routine diagnostic samples, an average rate of 29.95% (985/3289) of anti-N. caninum antibodies were detected. This rate was statistically higher than that of a previous study conducted in 2003, which reported a rate of 20% (p=0.01). Similar data were found in the fetal bovine serum samples, which showed an increase compared to previous studies conducted in 2010 that reported a rate of 15% (p=0.003). The increase in the detection rate of N. caninum antibodies underscores the need for measures to control and prevent bovine neosporosis.
Abstract in Portuguese:
Este estudo teve como objetivo determinar a frequência de detecção de anticorpos anti-Neospora caninum em três diferentes amostras e métodos de coleta: um estudo de prevalência, amostras de laboratório de diagnóstico de rotina e amostras de soro fetal bovino. Essas amostras foram coletadas de bovinos no Rio Grande do Sul (RS), região Sul do Brasil, e analisadas usando a técnica de reação de imunofluorescência indireta. Para cada método de coleta, um estudo histórico foi usado como referência para comparação. No estudo de prevalência, foram coletadas 1.248 amostras de soro entre 2020 e 2022. A prevalência de N. caninum no estado do RS foi de 22,8% (285/1248). Esse valor foi estatisticamente diferente quando comparado a estudos anteriores realizados em 2002, que relataram uma prevalência de 11,2% (p<0,001). Nas amostras de diagnóstico de rotina, foi detectada uma taxa média de 29,95% (985/3289) de anticorpos anti-N. caninum. Essa taxa foi estatisticamente maior do que a de um estudo anterior realizado em 2003, que relatou uma taxa de 20% (p=0,01). Dados semelhantes foram encontrados nas amostras de soro fetal bovino, que mostraram um aumento em comparação com estudos anteriores realizados em 2010 que relataram uma taxa de 15% (p=0,003). O aumento na taxa de detecção de anticorpos anti-N. caninum destaca a necessidade de medidas para controlar e prevenir a neosporose bovina.
Abstract in English:
Iguanids are susceptible to several bacterial infections, especially those caused by Gram-negative bacteria. The development of these diseases in reptiles is related to management, inadequate sanitary conditions, and immunosuppression. This study aims to describe the anatomopathological and microbiological aspects of a case of co-infection by Plesiomonas shigelloides, Citrobacter freundii and Aeromonas jandaei diagnosed in a free-living green iguana (Iguana iguana). Macroscopically, the lesions were mainly located in skeletal muscle, myocardium, small intestine, and liver, characterized by white-yellowish, multifocal, friable, irregular areas associated with necrosis and hemorrhage. In the histopathological analysis, basophilic bacillary structures corresponding to bacterial aggregates were observed in the skeletal muscle, myocardium, hepatic parenchyma, kidney, stomach and small intestine associated with areas of thrombosis, necrosis and hemorrhage. The diagnosis of sepsis by P. shigelloides, C. freundii and A. jandaei was confirmed by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) associated with the anatomopathological and microbiological findings observed in this case.
Abstract in Portuguese:
Iguanídeos são suscetíveis a diversas infecções bacterianas, principalmente aquelas causadas por bactérias Gram-negativas. O desenvolvimento destas enfermidades em répteis está relacionado a manejos, condições sanitárias inadequadas e imunossupressão. Este estudo tem por objetivo descrever os aspectos anatomopatológicos e microbiológicos de um caso de coinfecção por Plesiomonas shigelloides, Citrobacter freundii e Aeromonas jandaei diagnosticado em uma iguana verde de vida livre (Iguana iguana). Macroscopicamente, as lesões localizavam-se principalmente em musculatura esquelética, miocárdio, duodeno e fígado, caracterizando-se por áreas branco-amareladas, multifocais, friáveis, e irregulares associadas à necrose e hemorragia. No histopatológico foram vistas estruturas bacilares basofílicas correspondentes a agregados bacterianos em músculo esquelético, miocárdio, parênquima hepático, rim, estômago e intestino delgado associados a áreas de trombose, necrose e hemorragia. O diagnóstico de sepse por P. shigelloides, C. freundii e A. jandaei foi confirmado pela técnica de ionização por dessorção a laser assistida por matriz com analisador por tempo de voo (MALDI-TOF) associada aos achados anatomopatológicos e microbiológicos observados neste caso.