Clonality test by PCR - PARR in real time of canine lymphomas

Lymphoma is a neoplasm of hematopoietic origin that affects canines. The proper establishment of prognosis and rapid institution of treatment are essential for a better quality of life, and immunophenotyping is one of the tools used for this purpose. The objective of this study was to perform a clonality test for immunophenotypic characterization of canine lymphomas using the polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) technique in real-time from samples fixed in formalin and embedded in paraffin. The 23 analyzed samples were fixed in formalin and embedded in paraffin canine lymphoma from the collection Laboratory of Histopathology of the Animal Pathology Area of the Departament of Veterinary Medicine - Federal Rural University of Pernambuco (UFRPE). Samples were processed, their DNA was extracted, quantified, diluted, and standardized at a concentration of 50 ng/µL. After extraction, all samples were subjected to conventional PCR for endogenous control (detection of the IgM target region), in which the extracted DNA was amplified in a final volume of 25 µL. The 128 bp amplified product was detected by 1.5% agarose gel electrophoresis. Of the 23 samples analyzed for the detection of the conserved region referring to the endogenous gene, 91.30% (21/23) amplified the conserved region Cµ by conventional PCR, and two samples 8.70% (2/23) were negative. Endogenous control positive samples were subjected to real-time PCR-PARR for detection of IgH Major and IgH Minor for B lymphocytes (LB), and TCRy for lymphocytes T (LT) target regions. All reactions were performed in duplicate to reduce the risk of false-positive or false-negative results due to technical errors. Samples previously confirmed by immunohistochemistry were used as positive controls for T cell and B cell lymphoma, and MilliQ water was used as a negative reaction control. After amplification, the melting curve gradually increased the temperature by 1o C/5 s to 95o C during continuous fluorescence monitoring. Of the 21 samples analyzed, 100.00% (21/21) demonstrated clonal amplification. Of these, 57.15% (12/21) were positive for phenotype B, and 42.85% (9/21) were positive for phenotype T. Due to the importance of researching and confirming samples from files fixed and embedded in paraffin samples in laboratories, PCR-PARR is a good tool for this purpose. In the present study, real-time PCR analysis demonstrated greater sensitivity in the characterization of the immunophenotype of lymphomas from old samples fixed in formalin and embedded in paraffin. The temperature of melting curve analysis may vary depending on the amount of DNA and its quality. In the present study, it was found that the average melting temperature in the samples varied between ± 3o C when compared to that in the control sample for LB and LT, 83.5o C and 80o C, respectively: in the literature, there is a relative difference in this temperature, which may vary up to 4o C. Real-time PCR-PARR was satisfactory in the characterization of the immunophenotype of canine lymphomas from formalin-fixed and paraffin-embedded samples; therefore, its use is recommended for both retrospective studies. The use of PCR-PARR associated with histopathological and/or cytopathological examination in cases of canine lymphomas strongly helps pathologists, provide a safe establishment of the immunophenotype, minimize errors, and optimize the diagnosis, thus directly contributing to the establishment of the prognosis.(AU)

Intussuscepção gastroesofágica em canino / Gastroesophageal intussusception in canine

Background: Gastroesophageal intussusception is characterized by the invagination of the stomach into the esophagus,with or without the involvementof adjacent organs such as the spleen, pancreas, and omentum. In dogs, this condition hasno breed or sex predisposition. As it is an infrequent disease in routine veterinary medical practice, this study reports acase of gastroesophageal intussusception in a dog necropsied at the Veterinary Hospital of the Federal Rural Universityof Pernambuco (HOVET - UFRPE), Recife, Brazil.Case: The body of a 12-year-old black mixed breed male dog was sent to the Pathology Department (Necropsy Sectorof the Federal Rural University of Pernambuco - UFRPE) for a necropsy. The animal had a previous 4-year history ofrecurrent emesis and limb weakness, primarily in the anterior limbs, that worsened in the previous months and progressedto death. No previous treatments were reported by the owner. On external examination, the animal had a low body score(cachectic), forelimb joints with great flexibility, congested oral and ocular mucous membranes, enophthalmos, and increased volume in the perianal region. At the opening of the thoracic cavity, the final third of the esophagus was dilatedand gastroesophageal intussusception, edema, and pulmonary congestion were noted. In the abdominal cavity, there washepatic and renal congestion and large intestine and rectal ampoule dilation, with a large amount of solid and retainedfeces (fecaloma), perianal hernia, and testicular neoformation. These findings were consistent with those observed in deathcaused by cardiorespiratory failure secondary to gastroesophageal intussusception.Discussion: The pathophysiology of gastroesophageal intussusception is still not elucidated and is probably multifactorial.This condition causes reverse gastric peristalsis associated with a sudden and sustained increase in abdominal...

Intussuscepção gastroesofágica em canino / Gastroesophageal intussusception in canine

Background: Gastroesophageal intussusception is characterized by the invagination of the stomach into the esophagus,with or without the involvementof adjacent organs such as the spleen, pancreas, and omentum. In dogs, this condition hasno breed or sex predisposition. As it is an infrequent disease in routine veterinary medical practice, this study reports acase of gastroesophageal intussusception in a dog necropsied at the Veterinary Hospital of the Federal Rural Universityof Pernambuco (HOVET - UFRPE), Recife, Brazil.Case: The body of a 12-year-old black mixed breed male dog was sent to the Pathology Department (Necropsy Sectorof the Federal Rural University of Pernambuco - UFRPE) for a necropsy. The animal had a previous 4-year history ofrecurrent emesis and limb weakness, primarily in the anterior limbs, that worsened in the previous months and progressedto death. No previous treatments were reported by the owner. On external examination, the animal had a low body score(cachectic), forelimb joints with great flexibility, congested oral and ocular mucous membranes, enophthalmos, and increased volume in the perianal region. At the opening of the thoracic cavity, the final third of the esophagus was dilatedand gastroesophageal intussusception, edema, and pulmonary congestion were noted. In the abdominal cavity, there washepatic and renal congestion and large intestine and rectal ampoule dilation, with a large amount of solid and retainedfeces (fecaloma), perianal hernia, and testicular neoformation. These findings were consistent with those observed in deathcaused by cardiorespiratory failure secondary to gastroesophageal intussusception.Discussion: The pathophysiology of gastroesophageal intussusception is still not elucidated and is probably multifactorial.This condition causes reverse gastric peristalsis associated with a sudden and sustained increase in abdominal...(AU)

Intercurrence of paratuberculosis in intradermal tuberculin test reactive cattle

Background: Bovine tuberculosis control programs are based on a standard diagnostic method, the intradermal test with purified protein derivatives, which is used to identify and eliminate diseased animals. Currently none of the tests available allow complete differentiation between infected and uninfected animals. The main limitations of the tests available are related to diagnostic sensitivity and specificity, which results in false positive reactions due to the existence of cross infections, and also false negative, inherent to the state of energy of some animals. The aim of this work was to study the intercurrence of paratuberculosis in tuberculosis reactive cattle by the comparative cervical test. Materials, Methods & Results: Three hundred and thirty four cattle were evaluated using the comparative cervical test (CCT) and serology for tuberculosis (TB) and paratuberculosis (PTB) ELISA IDEXX®. All of the animals testing positive by CCT were euthanized and necropsied. Fragments of lymph node, lung and intestine were collected and analyzed using histopathological techniques, with staining by Hematoxylin-Eosin (HE). Samples of lung and lymph nodes (retropharyngeal, submandibular, cervical and mediastinal) of the animals testing positive by CCT were evaluated using qPRC for M. bovis, and intestinal and mesenteric lymph nodes using PCR for PTB. Of the 334 cattle evaluated using the comparative cervical test, 16 were considered positive. No lesions suggestive of tuberculosis were found in the macroscopic inspection of the carcasses. The most evident anatomical and pathological finding was a thickening of intestinal mucosa, found in 12 of the 16 cattle submitted to necropsy. No microscopic lesions suggestive of TB were identified nor was the presence of M. bovis detected by qPCR. The main histopathological findings were observed in the small intestine and mesenteric lymph nodes and identified as enteritis, lymphangitis...

Dilatação e torção de ceco associada à obstrução por fitobezoar em bezerra / Cecal dilatation and torsion associated with phytobezoar obstruction in a heifer

Background: Dilatation of the cecum associated with intestinal torsion is a fermentative digestive disease that has a lowoccurrence in veterinary medicine, particularly in large animal practice. This condition is most prevalent in dairy cattle.The etiology of this digestive system disorder is related to changes in nutritional management. As it is an uncommoncondition that is frequently observed in the practice of veterinary medicine, the objective of this report was to describe acase of dilatation and torsion of the cecum associated with phytobezoar obstruction in a heifer that was examined at theVeterinary Hospital of the Federal Rural University of Pernambuco (HOVET-UFRPE), Recife, Brazil.Case: A 55-day-old heifer, with a history of abdominal distension associated with difficulty feeding and neurological signsof hind limb paresis, intentional head tremors, and prostration, was examined at the HOVET-UFRPE large animal clinic.A clinical examination demonstrated the presence of enophthalmos, abdominal distension, pale coloration of the ocularmucous membranes, and a moderate level of dehydration (3%). The dehydration was treated through the administrationof 1.150 L of saline solution along with 6 mL of Bionew (Vetnil®), 15 mL of calcium solution, and 5 mL of 50% glucose.Subsequently, the animal underwent intestinal washing through the insertion of an oral probe, which resulted in the expulsion of putrid content. Afterward, the animal’s blood was collected and sent to the Clinical Pathology Laboratory of theDepartment of Veterinary Medicine at HOVET-UFRPE for a complete blood count...

Dilatação e torção de ceco associada à obstrução por fitobezoar em bezerra / Cecal dilatation and torsion associated with phytobezoar obstruction in a heifer

Background: Dilatation of the cecum associated with intestinal torsion is a fermentative digestive disease that has a lowoccurrence in veterinary medicine, particularly in large animal practice. This condition is most prevalent in dairy cattle.The etiology of this digestive system disorder is related to changes in nutritional management. As it is an uncommoncondition that is frequently observed in the practice of veterinary medicine, the objective of this report was to describe acase of dilatation and torsion of the cecum associated with phytobezoar obstruction in a heifer that was examined at theVeterinary Hospital of the Federal Rural University of Pernambuco (HOVET-UFRPE), Recife, Brazil.Case: A 55-day-old heifer, with a history of abdominal distension associated with difficulty feeding and neurological signsof hind limb paresis, intentional head tremors, and prostration, was examined at the HOVET-UFRPE large animal clinic.A clinical examination demonstrated the presence of enophthalmos, abdominal distension, pale coloration of the ocularmucous membranes, and a moderate level of dehydration (3%). The dehydration was treated through the administrationof 1.150 L of saline solution along with 6 mL of Bionew (Vetnil®), 15 mL of calcium solution, and 5 mL of 50% glucose.Subsequently, the animal underwent intestinal washing through the insertion of an oral probe, which resulted in the expulsion of putrid content. Afterward, the animals blood was collected and sent to the Clinical Pathology Laboratory of theDepartment of Veterinary Medicine at HOVET-UFRPE for a complete blood count...(AU)

Low prevalence of infection by Sarcocystis neurona in horses from the State of Alagoas, Brazil / Baixa prevalência da infecção por Sarcocystis neurona em cavalos do estado de Alagoas, Brasil

The aim of this study was to determine the prevalence of infection by Sarcocystis neurona in horses and identify potential risk factors. Were analyzed 427 samples from 36 farms in 21 municipalities in the Alagoas State, Brazil. Presence of anti-S. neurona antibodies was diagnosed by indirect immunofluorescence antibody test (IFAT) and was confirmed using the immunoblot test. Risk factors were assessed through investigative questionnaires on animal management on the farms. The prevalence of anti-S.neurona antibodies was 2.8% (confidence interval, CI: 1.5-4.9%) from IFAT and 1.6% (CI:0.8-3.34%) from immunoblot, and there were positive horses on 16.6% of the studied farms. None of the variables studied presented associations with serological status for S. neurona. This is the first report on infection by S. neurona in horses reared in Alagoas, Brazil showing a low exposure to S. neurona in this region, but with significant numbers of foci.(AU)

Objetivou-se neste estudo determinar a prevalência e os fatores de risco associados à infecção por Sarcocystis neurona em equinos. Foram analisadas 427 amostras de 36 propriedades localizadas em 21 municípios do estado de Alagoas. O diagnóstico de anticorpos anti-S. neurona foi realizado pela técnica de Imunofluorescência Indireta (IFI) e confirmada por immunoblot. O estudo dos fatores de risco foi realizado a partir de questionários investigativos sobre o manejo dos animais nas propriedades. A prevalência de anticorpos anti-S. neurona foi de 2,8% (I.C. 1,5-4,9%) na IFI e de 1,6% (I.C. 0,8-3,34%) no immunoblot com equinos positivos em 16,6% das propriedades estudadas. Nenhuma variável estudada apresentou associação com o status sorológico para S. neurona. Este é o primeiro relato da infecção por S. neurona em equinos criados no Estado de Alagoas, Brasil, confirmando que os animais desta região têm baixa exposição a S. neurona, mas com significativo número de focos.(AU)

Anti-Leptospira spp. antibodies in pigs slaughtered in the agreste region of Pernambuco, Brazil

Background: Swine production and productivity rates can be influenced by several factors, such as genetics, environmentalconditions, nutritional factors, previous infections and others. Among infectious diseases, leptospirosis is a well-knowncause of reproductive disorders in pigs. These animals are considered carriers of the disease when they are in the terminalstage of the infection and the Veterinary Inspection Service has not been notified when they are slaughtered. Consideringthe lack of epidemiological information on Leptospira infection in pigs in the state of Pernambuco, the aim of this studywas to investigate anti-Leptospira antibodies in pigs slaughtered in the Agreste region of the state of Pernambuco, Brazil.Materials, Methods & Results: Blood samples were collected from 305 pigs in 11 municipalities in the Agreste region ofPernambuco. The animals had no history of vaccination, and were raised on subsistence-oriented family farms. The serawere subjected to the Microscopic Agglutination Test (MAT) for the detection of antibodies. The serovars used in the MATwere: Icterohaemorrhagiae, Copenhageni, Javanica, Canicola, Castellonis, Pyrogenenes, Cynopteri, Autumnalis, Sentot,Djasiman, Australis, Pomona, Grippotyphosa, Hebdomadis, Wolffi, Sejroe, Saxkoebing, Bataviae, Tarassovi, Panama,Patoc, Andamana, Celledoni, Shermani, Brastilava and Hardjo. Sera showing titers of ≥100 were considered positive. TheMAT results indicated that 78/305 (25.57%) of the samples were positive, and were distributed in the 11 municipalities.The most frequent serovars were Icterohaemorrhagiae, Copenhageni, and Djasiman, with frequencies of 55.13%, 17.95%and 6.41%, respectively.Discussion: Swine infected with leptospirosis showed few or no signs of the disease. However, the bacteria can be carriedfor long periods in convoluted tubules of the kidney and their urinary excretion may last for years...

Melanoma em cabra leiteira / Melanoma in a dairy goat

Background: Skin neoplasms in goats, albeit less frequent than in equines and cattle, are no less important, given thatthe presence of melanoma is worrisome due to its local and metastatic aggressiveness, which leads to death, especially innortheastern Brazil, where ultraviolet (UV) radiation levels are high, being an important factor the development of thisneoplasia, especially in light-skinned animals. This report describes the clinical and pathological aspects, beyond the attempted surgical treatment a case of Saanen goat melanoma raised in Feira Nova, Pernambuco, Brazil.Case: A 4-year-old pregnant Saanen goat was treated at the AGA/UFRPE. The animal presented blackened fetid masseslocated on the lower lip and ventral region of the jaw that had grown for two months. In addition to the tumor, a physicalexamination revealed altered submandibular and left prescapular lymph nodes. Local anesthesia and sedation were applied and both neoplasms were excised. The goat was then medicated with anti-inflammatory and antibiotic drugs anddischarged. However, 45 days later, the goat returned with multiple nodules similar in appearance to the previous one andin a severe clinical condition, characterized by pale mucous membranes, dehydration, enophthalmia, bilateral nasal seroussecretion and enlargement of the lymph nodes mandibular, retro pharyngeal and right paratid, as well as slight respiratorysnores in the ventral portion of lungs and productive cough. Due to the unfavorable prognosis, the animal was euthanized.Macroscopic findings revealed multiple nodular, infiltrative and blackened lesions on the tongue, lung, kidneys and liver,enlarged and blackened lymph nodes, and sagittal section of the cervical spine identified a dark spot on the seventh thoracic vertebra. In the histological evaluation of necrotic lesions present in the skin, lung, liver and kidneys was observednodular...

Anti-Leptospira spp. antibodies in pigs slaughtered in the agreste region of Pernambuco, Brazil

Background: Swine production and productivity rates can be influenced by several factors, such as genetics, environmentalconditions, nutritional factors, previous infections and others. Among infectious diseases, leptospirosis is a well-knowncause of reproductive disorders in pigs. These animals are considered carriers of the disease when they are in the terminalstage of the infection and the Veterinary Inspection Service has not been notified when they are slaughtered. Consideringthe lack of epidemiological information on Leptospira infection in pigs in the state of Pernambuco, the aim of this studywas to investigate anti-Leptospira antibodies in pigs slaughtered in the Agreste region of the state of Pernambuco, Brazil.Materials, Methods & Results: Blood samples were collected from 305 pigs in 11 municipalities in the Agreste region ofPernambuco. The animals had no history of vaccination, and were raised on subsistence-oriented family farms. The serawere subjected to the Microscopic Agglutination Test (MAT) for the detection of antibodies. The serovars used in the MATwere: Icterohaemorrhagiae, Copenhageni, Javanica, Canicola, Castellonis, Pyrogenenes, Cynopteri, Autumnalis, Sentot,Djasiman, Australis, Pomona, Grippotyphosa, Hebdomadis, Wolffi, Sejroe, Saxkoebing, Bataviae, Tarassovi, Panama,Patoc, Andamana, Celledoni, Shermani, Brastilava and Hardjo. Sera showing titers of ≥100 were considered positive. TheMAT results indicated that 78/305 (25.57%) of the samples were positive, and were distributed in the 11 municipalities.The most frequent serovars were Icterohaemorrhagiae, Copenhageni, and Djasiman, with frequencies of 55.13%, 17.95%and 6.41%, respectively.Discussion: Swine infected with leptospirosis showed few or no signs of the disease. However, the bacteria can be carriedfor long periods in convoluted tubules of the kidney and their urinary excretion may last for years...(AU)

Melanoma em cabra leiteira / Melanoma in a dairy goat

Background: Skin neoplasms in goats, albeit less frequent than in equines and cattle, are no less important, given thatthe presence of melanoma is worrisome due to its local and metastatic aggressiveness, which leads to death, especially innortheastern Brazil, where ultraviolet (UV) radiation levels are high, being an important factor the development of thisneoplasia, especially in light-skinned animals. This report describes the clinical and pathological aspects, beyond the attempted surgical treatment a case of Saanen goat melanoma raised in Feira Nova, Pernambuco, Brazil.Case: A 4-year-old pregnant Saanen goat was treated at the AGA/UFRPE. The animal presented blackened fetid masseslocated on the lower lip and ventral region of the jaw that had grown for two months. In addition to the tumor, a physicalexamination revealed altered submandibular and left prescapular lymph nodes. Local anesthesia and sedation were applied and both neoplasms were excised. The goat was then medicated with anti-inflammatory and antibiotic drugs anddischarged. However, 45 days later, the goat returned with multiple nodules similar in appearance to the previous one andin a severe clinical condition, characterized by pale mucous membranes, dehydration, enophthalmia, bilateral nasal seroussecretion and enlargement of the lymph nodes mandibular, retro pharyngeal and right paratid, as well as slight respiratorysnores in the ventral portion of lungs and productive cough. Due to the unfavorable prognosis, the animal was euthanized.Macroscopic findings revealed multiple nodular, infiltrative and blackened lesions on the tongue, lung, kidneys and liver,enlarged and blackened lymph nodes, and sagittal section of the cervical spine identified a dark spot on the seventh thoracic vertebra. In the histological evaluation of necrotic lesions present in the skin, lung, liver and kidneys was observednodular...(AU)

Criptococose sistêmica em um canino da raça Miniatura Schnauzer / Systemic cryptococcosis in a Miniature Schnauzer dog

Background: Cryptococcosis is an important zoonotic disease that occurs worldwide. The disease is caused by a soilborne opportunistic fungus of the genus Cryptococcus, which can also be found in the feces of birds, especially pigeons. In Brazil, the geographical distribution of the agent is fairly defined, with the species C. gattii predominantly found in the north-eastern states. Diagnosis is based on the clinical history, physical examination findings, and results of complementary testing, such as cytopathological and histopathological examinations. This report aims to describe the clinical and anatomopathological findings in a case of systemic cryptococcosis in a dog.Case: A 4-year-old female Miniature Schnauzer was referred for necropsy. For 3 months prior to death, the dog had displayed generalized alopecia, pruritus, and severe weight loss, Initially, a private veterinarian had suspected leishmaniasis. Gross examination was performed and samples for cytopathology, histopathology, and fungal culture were obtained. The macroscopic examination revealed generalized alopecia, congested mucosa, cachexia, hyperemia of the brain, and multiple white nodules measuring 0.5 cm to 4 cm in the lungs and the kidneys. None of the other organs showed significant lesions. Cytopathological examination of the nodules revealed a highly cellular sample, with a severe chronic inflammatory response, characterized by multinucleated giant cells and round-to-elliptical, yeast-shaped structures (5-10 µm), suggestive of Cryptococcus organisms. Histopathological examination of the lungs, kidneys, and brain revealed a severe diffuse chronic inflammatory process, with lymphocytic infiltration and multinucleated giant cells; countless yeast-shaped, round-to-ovoid structures (similar in appearance to “soap bubbles”) that were negative in hematoxylin-eosin stain were also present.[...]

Criptococose sistêmica em um canino da raça Miniatura Schnauzer / Systemic cryptococcosis in a Miniature Schnauzer dog

Background: Cryptococcosis is an important zoonotic disease that occurs worldwide. The disease is caused by a soilborne opportunistic fungus of the genus Cryptococcus, which can also be found in the feces of birds, especially pigeons. In Brazil, the geographical distribution of the agent is fairly defined, with the species C. gattii predominantly found in the north-eastern states. Diagnosis is based on the clinical history, physical examination findings, and results of complementary testing, such as cytopathological and histopathological examinations. This report aims to describe the clinical and anatomopathological findings in a case of systemic cryptococcosis in a dog.Case: A 4-year-old female Miniature Schnauzer was referred for necropsy. For 3 months prior to death, the dog had displayed generalized alopecia, pruritus, and severe weight loss, Initially, a private veterinarian had suspected leishmaniasis. Gross examination was performed and samples for cytopathology, histopathology, and fungal culture were obtained. The macroscopic examination revealed generalized alopecia, congested mucosa, cachexia, hyperemia of the brain, and multiple white nodules measuring 0.5 cm to 4 cm in the lungs and the kidneys. None of the other organs showed significant lesions. Cytopathological examination of the nodules revealed a highly cellular sample, with a severe chronic inflammatory response, characterized by multinucleated giant cells and round-to-elliptical, yeast-shaped structures (5-10 µm), suggestive of Cryptococcus organisms. Histopathological examination of the lungs, kidneys, and brain revealed a severe diffuse chronic inflammatory process, with lymphocytic infiltration and multinucleated giant cells; countless yeast-shaped, round-to-ovoid structures (similar in appearance to “soap bubbles”) that were negative in hematoxylin-eosin stain were also present.[...](AU)

Detection of Mycobacterium avium subsp. paratuberculosis in bovine milk from the state of Pernambuco, Brazil

The aim of this study was to detect the IS900 region of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine milk samples using real-time polymerase chain reaction (qPCR) and conventional PCR, and to study the agreement between these tests. A total of 121 bovine milk samples were collected from herds considered positive for MAP, from the State of Pernambuco, Brazil. MAP DNA was detected in 20 samples (16.5%) using conventional PCR and in 34 samples (28.1%) using qPCR. MAP DNA was detected in all of the 6 animal farms studied. Moderate agreement was found between qPCR and conventional PCR results, where the sensitivity and specificity of conventional PCR in relation to qPCR were 50% and 96.6%, respectively. Thus, the IS900 region of MAP was found in bovine milk samples from the State of Pernambuco. To the best of our knowledge, this is the first report of MAP DNA found in bovine milk in Northeast Brazil. We also demonstrated the qPCR technique is more sensitive than conventional PCR with respect to detection of MAP in milk samples.(AU)

Occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chicken breeding in Northeast, Brazil / Ocorrência de anticorpos anti-Toxoplasma gondii e DNA do parasita em galinhas de criações domésticas no Nordeste, Brasil

The aim of the present study was to investigate the occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chickens bred in the metropolitan area of Recife, Brazil. In total, 212 serum samples were collected from 16 properties, and 12 backyard chickens were collected in the six sanitary districts of Recife. An indirect immunofluorescence assay (IFA) was used to investigate the occurrence of anti-Toxoplasma gondii antibodies. Polymerase chain reaction (PCR) was used to detect T. gondii DNA in brain, heart, liver and lung specimens. Of the samples analyzed by serology, 86/212 (40.56%) were positive; of the samples analyzed by PCR, 2/12 (16.7%) were positive, with both samples positive by both tests (serological and molecular). The presence of antibody anti-T. gondii and parasite DNA in tissues of these animals are worrying aspects for public health because there is a risk of transmission of the parasite to humans through eating undercooked or raw meat. Based on the results, the adoption of preventive measures to prevent the cats access to the chickens creations should be encouraged, since these animals were identified in most of the studied properties.(AU)

O objetivo do presente estudo foi investigar a ocorrência de anticorpos anti-Toxoplasma gondii e de DNA do parasito em galinhas de criações domésticas, na região metropolitana de Recife, Brasil. No total, 212 amostras de soro foram coletadas de aves de 16 estabelecimentos e de 12 galinhas de criações domésticas nos seis distritos sanitários de Recife. Para a pesquisa de anticorpos anti-Toxoplasma gondii foi utilizada a Reação de Imunofluorescência Indireta (RIFI). A Reação em Cadeia da Polimerase (PCR) foi utilizada para detectar o DNA de T. gondii em fragmentos de cérebro, coração, fígado e pulmão. Das amostras analisadas por sorologia, 86/212 (40,56%) foram positivas. Das amostras analisadas por PCR, 2/212 (16,7%) foram positivas, em ambos os testes (sorológicos e moleculares). A presença de anticorpos anti-T. gondii e de DNA parasitário nos tecidos desses animais são aspectos preocupantes para saúde pública, porque há o risco de transmissão do parasita para humanos através da ingestão de carne mal cozida ou crua. Com base nos resultados obtidos, a adoção de medidas preventivas que evitem o acesso de gatos às criações de galinhas deve ser incentivada, uma vez que esses animais foram identificados na maioria das propriedades estudadas.(AU)

Occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chicken breeding in Northeast, Brazil

Abstract The aim of the present study was to investigate the occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chickens bred in the metropolitan area of Recife, Brazil. In total, 212 serum samples were collected from 16 properties, and 12 backyard chickens were collected in the six sanitary districts of Recife. An indirect immunofluorescence assay (IFA) was used to investigate the occurrence of anti-Toxoplasma gondii antibodies. Polymerase chain reaction (PCR) was used to detect T. gondii DNA in brain, heart, liver and lung specimens. Of the samples analyzed by serology, 86/212 (40.56%) were positive; of the samples analyzed by PCR, 2/12 (16.7%) were positive, with both samples positive by both tests (serological and molecular). The presence of antibody anti-T. gondii and parasite DNA in tissues of these animals are worrying aspects for public health because there is a risk of transmission of the parasite to humans through eating undercooked or raw meat. Based on the results, the adoption of preventive measures to prevent the cats access to the chickens creations should be encouraged, since these animals were identified in most of the studied properties.

Resumo O objetivo do presente estudo foi investigar a ocorrência de anticorpos anti-Toxoplasma gondii e de DNA do parasito em galinhas de criações domésticas, na região metropolitana de Recife, Brasil. No total, 212 amostras de soro foram coletadas de aves de 16 estabelecimentos e de 12 galinhas de criações domésticas nos seis distritos sanitários de Recife. Para a pesquisa de anticorpos anti-Toxoplasma gondii foi utilizada a Reação de Imunofluorescência Indireta (RIFI). A Reação em Cadeia da Polimerase (PCR) foi utilizada para detectar o DNA de T. gondii em fragmentos de cérebro, coração, fígado e pulmão. Das amostras analisadas por sorologia, 86/212 (40,56%) foram positivas. Das amostras analisadas por PCR, 2/212 (16,7%) foram positivas, em ambos os testes (sorológicos e moleculares). A presença de anticorpos anti-T. gondii e de DNA parasitário nos tecidos desses animais são aspectos preocupantes para saúde pública, porque há o risco de transmissão do parasita para humanos através da ingestão de carne mal cozida ou crua. Com base nos resultados obtidos, a adoção de medidas preventivas que evitem o acesso de gatos às criações de galinhas deve ser incentivada, uma vez que esses animais foram identificados na maioria das propriedades estudadas.

Paratuberculose em bovinos de corte na região Sul do Rio Grande do Sul / Paratuberculosis in beef cattle in Southern Brazil

Descrevem-se os aspectos epidemiológicos e clínico-patológicos de paratuberculose diagnosticada no sul do Rio Grande do Sul em uma propriedade de bovinos de corte. Dois bovinos criados extensivamente que apresentavam emagrecimento progressivo e diarreia crônica foram necropsiados. Os linfonodos mesentéricos estavam aumentados e edematosos. A mucosa do intestino estava espessada e enrugada com aspecto cerebroide principalmente na porção final do íleo, válvula íleo-cecal e ceco. Fragmentos dos órgãos foram fixados em formalina 10%, incluídos em parafina, cortados e corados pela técnica de hematoxilina e eosina (HE) e Ziehl-Neelsen (ZN). Fezes foram encaminhadas ao Departamento de Medicina Veterinária, Área de Medicina Veterinária Preventiva da Universidade Federal Rural de Pernambuco para o cultivo de Mycobacterium aviumsubsp.paratuberculosis em meio Lowenstein Jensen com micobactina e para realização da PCR. Histologicamente, havia enterite granulomatosa no jejuno, íleo, ceco e reto, afetando multifocalmente, também, o duodeno e o cólon. Havia, ainda, linfangite e adenite granulomatosa. Pela coloração de ZN foram observados numerosos bacilos álcool-ácido resistentes (BAAR) no interior de macrófagos, células gigantes de Langhans e nos linfonodos mesentéricos no jejuno, íleo ceco e reto. Não houve crescimento bacteriano nas amostras de fezes e cinco amostras amplificaram a sequência genética IS900 específica do Mycobacterium aviumsubesp. paratuberculosis. Pelo presente trabalho pode-se concluir que a paratuberculose apesar dos poucos relatos ocorre também em bovinos de corte criados extensivamente no sul do Rio Grande do Sul e pode ter uma prevalência maior do que se supõe na região. Alerta-se para a necessidade do diagnóstico e da tomada de medidas efetivas de controle para esta doença que, por muitos, ainda é considerada uma doença exótica no Brasil.(AU)

The epidemiological, clinical and pathological aspects of paratuberculosis diagnosed in southern Rio Grande do Sul in a beef cattle property are described. Two cattle raised extensively and that presented progressive weight loss and chronic diarrhea were necropsied. The mesenteric lymph nodes were enlarged and edematous. The intestinal mucosa was thickened and wrinkled with cerebroid aspect, especially in the final portion of the ileum, ileocecal valve and cecum. Fragments of the organs were fixed in 10% formalin, embedded in paraffin, sectioned and stained with hematoxylin and eosin (HE) and Ziehl-Neelsen (ZN). Feces samples were referred to Departamento de Medicina Veterinária, Área de Medicina Veterinária Preventiva da Universidade Federal Rural de Pernambuco, for the cultivation of Mycobacterium avium subsp. paratuberculosis with Lõwenstein Jensen for mycobacterial and PCR analysis. Histologically, granulomatous enteritis was observed in the jejunum, ileum, cecum and rectum, and in multifocal areas the duodenum and colon. Lymphangitis and granulomatous adenitis was also observed. In ZN staining numerous resistant acid-fast bacilli (AFB) within macrophages, giant cells of Langhans and mesenteric lymph nodes in the jejunum, ileum cecum and rectum were observed. There was no bacterial growth in stool samples. Five samples amplified the gene sequence IS900 specific for Mycobacterium aviumsubsp. paratuberculosis. It can be concluded that paratuberculosis, despite the few reports, occurs in beef cattle raised extensively in southern Rio Grande do Sul, Paratuberculosis may have a higher prevalence in the region than it is assumed. There is need for diagnosis of this disease and to assume effective measures for its control, as for many it is still considered an exotic malady in Brazil.(AU)

Occurrence of anti-Neospora caninum IgG antibodies in dairy cows in the state of Pernambuco, Brazil / Ocorrência de anticorpos IgG anti-Neospora caninum em vacas leiteiras no estado de Pernambuco, Brasil

Neosporosis is a disease caused by Neospora caninum, considered a major cause of abortion, responsible for major economic losses and sanitary problems in cattle creations. This study was performed to search for anti-Neospora caninum IgG antibodies in dairy cows. Blood serum samples were obtained from 533 cows from 13 municipalities in the state of Pernambuco. For the detection of antibodies was used the immunofluorescent antibody test (IFAT). IFAT was possible to determine a positivity of 6.4% (34/533). The results demonstrate the presence of infection by Neospora caninum in different regions of the state of Pernambuco.(AU)

A Neosporose é uma enfermidade causada pelo Neospora caninum, considerada uma importante causa de aborto, responsável por sérios prejuízos econômicos e sanitários em criações de bovinos. Neste estudo foi realizada a pesquisa de anticorpos IgG anti-Neospora caninum em vacas leiteiras. Foram obtidas amostras de soro sanguíneo de 533 vacas provenientes de 13 municípios no Estado de Pernambuco. Para a pesquisa de anticorpos foi empregada a técnica de Imunofluorescência Indireta (RIFI). Na RIFI foi possível determinar uma positividade de 6,4% (34/533). Os resultados obtidos demostram a presença da infecção por Neospora caninum em diferentes regiões do Estado de Pernambuco.(AU)

Occurrence of anti-Neospora caninum IgG antibodies in dairy cows in the state of Pernambuco, Brazil / Ocorrência de anticorpos IgG anti-Neospora caninum em vacas leiteiras no estado de Pernambuco, Brasil

Neosporosis is a disease caused by Neospora caninum, considered a major cause of abortion, responsible for major economic losses and sanitary problems in cattle creations. This study was performed to search for anti-Neospora caninum IgG antibodies in dairy cows. Blood serum samples were obtained from 533 cows from 13 municipalities in the state of Pernambuco. For the detection of antibodies was used the immunofluorescent antibody test (IFAT). IFAT was possible to determine a positivity of 6.4% (34/533). The results demonstrate the presence of infection by Neospora caninum in different regions of the state of Pernambuco.

A Neosporose é uma enfermidade causada pelo Neospora caninum, considerada uma importante causa de aborto, responsável por sérios prejuízos econômicos e sanitários em criações de bovinos. Neste estudo foi realizada a pesquisa de anticorpos IgG anti-Neospora caninum em vacas leiteiras. Foram obtidas amostras de soro sanguíneo de 533 vacas provenientes de 13 municípios no Estado de Pernambuco. Para a pesquisa de anticorpos foi empregada a técnica de Imunofluorescência Indireta (RIFI). Na RIFI foi possível determinar uma positividade de 6,4% (34/533). Os resultados obtidos demostram a presença da infecção por Neospora caninum em diferentes regiões do Estado de Pernambuco.

Seroprevalence of antibodies to Neospora caninum in dogs in the state of Alagoas, Brazil / Soroprevalência de anticorpos anti-Neospora caninum em cães no estado de Alagoas, Brasil

This study investigated occurrences of antibodies to anti-Neospora caninum in dogs in the state of Alagoas, Brazil, and the risk factors associated with the infection. To investigate antibodies against N. caninum, an indirect immunofluorescence antibody test (IFAT) was used. To identify risk factors associated with infection by N. caninum, univariate analyses on variables of interest were conducted. The study analyzed 237 samples from dogs of both sexes and different breeds and ages, both from rural and from urban areas. Ten animals (4.2%) were seropositive: five (4.8%) from rural areas and five (3.8%) from urban areas. Regarding sex, five (4.0%) were male and five (4.5%) were female. In relation to breed, four animals of undefined breed and six of defined breed were positive. The ages of the seropositive animals ranged from two months to eleven years. Results show there are no significant associations between seropositivity and the variables analyzed. Results indicate neosporosis must be included in differential diagnosis of dog neurological disorders.

Investigou-se a ocorrência de anticorpos anti-Neospora caninum em cães no Estado de Alagoas, Brasil e os fatores de risco associados à infecção. Para a pesquisa de anticorpos contra N. caninum empregou-se a Reação de Imunofluorescênica Indireta (RIFI). Para identificar os fatores de risco associados à infecção pelo N. caninum foi realizada análise univariada das variáveis de interesse. Foram analisadas 237 amostras de cães de ambos os sexos, diferentes raças e idades e procedentes de áreas urbanas e rurais. Obteve-se 10 (4,2%) animais soropositivos, sendo cinco (4,8%) de áreas rurais e cinco (3,8%) de áreas urbanas. Com relação ao sexo, cinco (4,0%) eram machos e cinco (4,5%) fêmeas. Quanto às raças foram positivos quatro animais de raças indefinidas e seis de raças definidas. As idades dos animais soropositivos variaram de dois meses a 11 anos. Os resultados da análise estatística revelaram não haver associação significativa entre a soropositividade e as variáveis analisadas. Os resultados indicam a necessidade de incluir a neosporose no diagnóstico diferencial das desordens neurológicas caninas.