Presence of BRCA1 Gene Mutation in Bitches with Malignant Mammary Tumors

Background: Mammary tumors (MTs) in bitches are similar to breast cancers in women. Thus, they can be used as a model for human breast cancer and findings can be extrapolated for use in human medicine. BRCA1 is a tumor suppressor gene. When the gene has a mutation, it cannot repair damaged DNA, which causes genetic instability and tumorigenesis. Therefore, we aimed to study the frequency of single nucleotide polymorphisms (SNPs) in the BRCA1 gene that are associated with distinct histological types of malignant MT in bitches.Materials, Methods & Results: The study population consisted of 91 bitches, including a control group of 6 animals with healthy mammary glands and 85 animals with MTs. All animals underwent a presurgery evaluation consisting of a questionnaire administered to the person responsible for the animal, a physical examination, collection of peripheral blood for hematological and serum biochemistry evaluations, an electrocardiogram, and a preanesthesia evaluation. In addition, distant metastasis was studied via chest radiography and abdominal ultrasound. After evaluations were complete, the animals that could undergo surgery were administered general anesthesia and underwent a mastectomy or mammary gland sample collection. Histopathological examination and molecular analysis were performed to identify mutations in the BRCA1 gene. Histopathological examinations found 10 different types of malignant tumors in 36 sick animals. Tumor samples plus samples from the 6 control animals were subjected to DNA extraction, polymerase chain reaction (PCR) analysis, and genetic sequencing. The tumor with the highest incidence (33.33%) was a complex carcinoma, followed by carcinoma in mixed tumor (13.88), tubular carcinoma (13.88) and carcinosarcoma (13.88). Molecular analysis revealed 3 different SNP points in 5 samples (4006G>A, 3619A>G, and 3761C>T). The allelic variant 4006G>A (1/36) resulted in the alteration of the amino acid valine by isoleucine (V1336 I). The mutation 3619A>G (2/36) inserted the amino acid alanine instead of threonine (T1207 A). The mutation 3761C>T (2/36) led to the alteration of the amino acid serine by phenylalanine (S1254 F), a mutation for which there are no published reports. The histological types that showed BRCA1mutations were complex carcinoma (1/5), carcinoma in mixed tumor (1/5), papillary carcinoma (1/5) and tubular carcinoma (2/5). Software analysis identified the new SNP (nucleotide 3761) in BRCA1 and 2 points mutations in nucleotides 4006 and 3619 and responsible for genetic instability.Discussion: The development of breast cancer is caused by many endogenous and exogenous factors. The results of our study show that these factors have a greater presence in female, mixed breed, uncastrated, and older dogs, confirming the data in the veterinary literature. In the present study, we found different histological types of malignant breast tumors with mutations in the BRCA1 gene, as other authors have reported. However, we also found the mutation 3761C>T, which, to the best of our knowledge, has not been reported in the literature. This shows the need for studies in veterinary medicine that assess mutations in the BRCA1 gene and the most common histological types. In conclusion, SNPs in the BRCA1 gene cause genetic instability, resulting in additional mutations that lead to the development of breast tumors. They are point mutations that affect transcription, resulting in truncated proteins. These proteins may have a loss of function, leading to carcinogenesis.Keywords: polymorphism, BRCA1 gene, cancer, dogs.Descritores:polimorfismo, gene BRCA1, câncer, cães. Título: Presença da mutação do gene BRCA1 em cadelas com tumores mamários malignos.

Hemipelvectomia total em um gato com osteossarcoma osteoblástico

Background: Osteosarcoma (OSA) is a malignant neoplasm of mesenchymal tissue that exhibits various histological features and a differentiated biological clinical behavior. The combination of clinical, radiological, and histopathological features is important for diagnosis. Surgical treatment of neoplasms with pelvic involvement is a challenge. Primary bone tumors are rare in cats and are mostly reported in older animals. The objective of this paper was to report the case of a feline subjected to total hemipelvectomy as primary treatment of pelvic OSA; the surgical technique and histological analysis of the tumor are described. No signs of recurrence were seen during seven months of follow-up.Case: A 4-year-old female mixed breed cat weighing 3 kg was presented with a one year history of increased volume on the hip joint region. Palpation of the increased volume on the hip showed a firm, adhered, and painful mass. Radiographic examination of the pelvis in the dorso ventral projection exhibited a severe osteoblastic bone reaction on the proximal femur, as well as marked periosteal reaction of the hip joint with involvement of the acetabulum. A ventral approach total hemipelvectomy technique was adopted for excision of the tumor. The animal was placed on a lateral position and a circumferential skin incision was performed on the ipsilateral limb of the affected hemipelvis, on the medial portion of the femur, and was extended cranially up to the thoracolumbar region. The pubic symphysis was exposed with a periosteal elevator and a pubic symphysiotomy was performed on the midline using a Liston bone cutter; next, the sacroiliac junction was resected. No drain was used after surgery, only compressive bandages. The patient was hospitalized for post-operative monitoring which included opioids for pain management and a non-steroidal anti-inflammatory drug and a cephalosporin class antibiotic. The wound was cleaned with saline, rifamycin spray (Rifocina Spray®) was applied, and dressing was changed every 12 h. The excised bone was submitted for pathological analysis. Gross examination revealed a 4.2 cm hardened increase in volume on the area of the femoral head and acetabulum, with irregular surface; cut surface was hard and white with black and red spots. Histology revealed osteoblastic OSA with intense and solid fusiform and stellate cells distributed in a solid pattern with no capsular border, with marked infiltration of the bone tissue, marked osteolysis, moderate proliferation of osteoclasts, and moderate anisocytosis and anisokaryosis.Discussion: Bone tumors in cats are rare and develop spontaneously. They exhibit aggressive behavior and promote local bone destruction. Firm consistency and local swelling usually occur in more advanced cases when there is a delay between the onset of clinical signs and the veterinary consultation. Radiological findings such as bone destruction and bone formation are commonly seen in this type of tumor. Hemipelvectomy is considered an aggressive surgical technique, but necessary in many cases, especially when tumors are located in the pelvis and nearby tissues. In the case presented here, this surgery was the most viable and advised therapeutic option because of tumor location. No incidents (i.e. hemorrhage) occurred during the perioperative period. After surgery, the patient continued to perform all custom functions. The hind limb, which was affected in this case, is commonly reported to be affected in the literature; however, it is unusual to have involvement of both bones (pelvis and femur) since vascularization of these bones are independent. Diagnosis was confirmed by histopathology in which a characteristic malignant mesenchymal tumor of primitive bone cells was present, with production of osteoid of variable shapes and quantity with the presence of osteoclastic giant cells

Hemipelvectomia total em um gato com osteossarcoma osteoblástico

Background: Osteosarcoma (OSA) is a malignant neoplasm of mesenchymal tissue that exhibits various histological features and a differentiated biological clinical behavior. The combination of clinical, radiological, and histopathological features is important for diagnosis. Surgical treatment of neoplasms with pelvic involvement is a challenge. Primary bone tumors are rare in cats and are mostly reported in older animals. The objective of this paper was to report the case of a feline subjected to total hemipelvectomy as primary treatment of pelvic OSA; the surgical technique and histological analysis of the tumor are described. No signs of recurrence were seen during seven months of follow-up.Case: A 4-year-old female mixed breed cat weighing 3 kg was presented with a one year history of increased volume on the hip joint region. Palpation of the increased volume on the hip showed a firm, adhered, and painful mass. Radiographic examination of the pelvis in the dorso ventral projection exhibited a severe osteoblastic bone reaction on the proximal femur, as well as marked periosteal reaction of the hip joint with involvement of the acetabulum. A ventral approach total hemipelvectomy technique was adopted for excision of the tumor. The animal was placed on a lateral position and a circumferential skin incision was performed on the ipsilateral limb of the affected hemipelvis, on the medial portion of the femur, and was extended cranially up to the thoracolumbar region. The pubic symphysis was exposed with a periosteal elevator and a pubic symphysiotomy was performed on the midline using a Liston bone cutter; next, the sacroiliac junction was resected. No drain was used after surgery, only compressive bandages. The patient was hospitalized for post-operative monitoring which included opioids for pain management and a non-steroidal anti-inflammatory drug and a cephalosporin class antibiotic. The wound was cleaned with saline, rifamycin spray (Rifocina Spray®) was applied, and dressing was changed every 12 h. The excised bone was submitted for pathological analysis. Gross examination revealed a 4.2 cm hardened increase in volume on the area of the femoral head and acetabulum, with irregular surface; cut surface was hard and white with black and red spots. Histology revealed osteoblastic OSA with intense and solid fusiform and stellate cells distributed in a solid pattern with no capsular border, with marked infiltration of the bone tissue, marked osteolysis, moderate proliferation of osteoclasts, and moderate anisocytosis and anisokaryosis.Discussion: Bone tumors in cats are rare and develop spontaneously. They exhibit aggressive behavior and promote local bone destruction. Firm consistency and local swelling usually occur in more advanced cases when there is a delay between the onset of clinical signs and the veterinary consultation. Radiological findings such as bone destruction and bone formation are commonly seen in this type of tumor. Hemipelvectomy is considered an aggressive surgical technique, but necessary in many cases, especially when tumors are located in the pelvis and nearby tissues. In the case presented here, this surgery was the most viable and advised therapeutic option because of tumor location. No incidents (i.e. hemorrhage) occurred during the perioperative period. After surgery, the patient continued to perform all custom functions. The hind limb, which was affected in this case, is commonly reported to be affected in the literature; however, it is unusual to have involvement of both bones (pelvis and femur) since vascularization of these bones are independent. Diagnosis was confirmed by histopathology in which a characteristic malignant mesenchymal tumor of primitive bone cells was present, with production of osteoid of variable shapes and quantity with the presence of osteoclastic giant cells

Feline subcutaneous pythiosis

ABSTRACT: Pythiosis in felines is a rare disease associated with the oomycete Pythium insidiosum. The aim of this report was to describe the macroscopic, histopathological, and molecular characteristics of P. insidiosum infection in a 2-year-old cat, with a localized invasive subcutaneous mass. The feline had an increase of volume near the anal region since it was younger. The cat died just after surgery. The necropsy was performed, and samples were collected for histopathological examination. Microscopically, the skin lesion was characterized by necro-eosinophilic dermatitis, panniculitis, and myositis surrounding negatively stained hyphal structures. In the sections stained with GMS, dark brown hyphae were clearly seen inside the affected tissue. They were rarely septate and their walls were almost parallel. Immunohistochemistry using a polyclonal anti-P. insidiosum antibody showed a strongly immunostained hyphae into the lesions. The analysis based on PCR had a positive result for P. insidiosum. Pythiosis should be considered in the differential diagnosis of subcutaneous tissue disorders in felines

Periodontopathogens Detection in White-Cheeked Spider Monkey (Ateles marginatus)

Background: Control of oral lesions contributes directly to the survival, and welfare of captive animals, and studies show that the genus Ateles has a higher prevalence of widespread periodontal disease compared to other genera. Anaerobic microbial species, considered as periodontal pathogens, are part of the biofilm community that contributes to the development of periodontitis. The present study aimed to detect periodontopathogenos in the oral cavity of two captive white-cheeked spider monkeys (Ateles marginatus) submitted for assessment oral and subgingival curettage.Case: We evaluated one pair of captive white-cheeked spider monkeys, one male (A) and one female (B), of 15 years of age with an average weight of 7 kg. Animals were fed daily with rations for primates, including fruit, vegetables, and raw eggs. The animals underwent oral evaluation, and following the charting of odontogram and photographic documentation, both were classified with periodontal disease stage III, according to the AVDC (American College of Veterinary Dentistry). They presented with moderate periodontitis, characterized by a loss of 25 to 50% of periodontal insertion and exposure of furcation degree 2, measured through clinical survey. During intraoral review, animals underwent subgingival curettage with curette of Gracey on the surface of the canine vestibular (C) and four top bilateral premolars (4PM). Antibiotics were not used at the time of collection, for dealing with routine procedures of clinical evaluation. The animals showed an increase in the volume of hemorrhagic features in the vestibular region between C and the second pre molar (2PM) on the upper right. Incisional biopsy was collected immediately at the end of the assessment, for the purpose of histopathological analyses. The samples from subgingival collection were immediately deposited in microtubes containing 500 µL of 0.9% saline solution and kept at -18°C until the time of genomic DNA extraction. The extracted DNA were subjected to polymerase chain reaction (PCR) for the detection of Fusobacterium spp., Porphyromonas gingivalis, Prevotella intermedia and Tannerella forsythia. All the samples detected in PCR were confirmed by partial sequencing of genes. Fusobacterium spp., P. gingivalis, and T. forsythia were detected in both individuals A and B. The identification of P. intermedia was exclusive to animal A and histopathological analysis of the gingival fragment showed severe and diffuse neutrophilic infiltrates, confirming the of gingivitis for this individual, with greater severity in the disease progression.Discussion: Older animals in captivity have a higher incidence of periodontal disease and high expression of inflammatory cytokines and immune receptors involved in the induction of inflammation as compared to younger animals, as well as free-living animals. Gingivitis and periodontitis are associated with an increase in the frequency of anaerobic, gramnegative microorganisms, whereby the production of pigments and a variety of enzymes damages the periodontal tissues. Knowing that the presence of bacterial species directly affects the periodontal condition and that the participation of these microorganisms favors the installation and progression of the disease, this study describes the first molecular detection of periodontopathogens related to periodontal disease in white-cheeked spider monkeys. We concluded that these pathogens are present in the oral cavity of white-cheeked spider monkeys, highlighting the importance of routine and proper food handling in captive animals

Presence of BRCA1 Gene Mutation in Bitches with Malignant Mammary Tumors

Background: Mammary tumors (MTs) in bitches are similar to breast cancers in women. Thus, they can be used as a model for human breast cancer and findings can be extrapolated for use in human medicine. BRCA1 is a tumor suppressor gene. When the gene has a mutation, it cannot repair damaged DNA, which causes genetic instability and tumorigenesis. Therefore, we aimed to study the frequency of single nucleotide polymorphisms (SNPs) in the BRCA1 gene that are associated with distinct histological types of malignant MT in bitches.Materials, Methods & Results: The study population consisted of 91 bitches, including a control group of 6 animals with healthy mammary glands and 85 animals with MTs. All animals underwent a presurgery evaluation consisting of a questionnaire administered to the person responsible for the animal, a physical examination, collection of peripheral blood for hematological and serum biochemistry evaluations, an electrocardiogram, and a preanesthesia evaluation. In addition, distant metastasis was studied via chest radiography and abdominal ultrasound. After evaluations were complete, the animals that could undergo surgery were administered general anesthesia and underwent a mastectomy or mammary gland sample collection. Histopathological examination and molecular analysis were performed to identify mutations in the BRCA1 gene. Histopathological examinations found 10 different types of malignant tumors in 36 sick animals. Tumor samples plus samples from the 6 control animals were subjected to DNA extraction, polymerase chain reaction (PCR) analysis, and genetic sequencing. The tumor with the highest incidence (33.33%) was a complex carcinoma, followed by carcinoma in mixed tumor (13.88), tubular carcinoma (13.88) and carcinosarcoma (13.88). Molecular analysis revealed 3 different SNP points in 5 samples (4006G>A, 3619A>G, and 3761C>T). The allelic variant 4006G>A (1/36) resulted in the alteration of the amino acid valine by isoleucine (V1336 I). The mutation 3619A>G (2/36) inserted the amino acid alanine instead of threonine (T1207 A). The mutation 3761C>T (2/36) led to the alteration of the amino acid serine by phenylalanine (S1254 F), a mutation for which there are no published reports. The histological types that showed BRCA1mutations were complex carcinoma (1/5), carcinoma in mixed tumor (1/5), papillary carcinoma (1/5) and tubular carcinoma (2/5). Software analysis identified the new SNP (nucleotide 3761) in BRCA1 and 2 points mutations in nucleotides 4006 and 3619 and responsible for genetic instability.Discussion: The development of breast cancer is caused by many endogenous and exogenous factors. The results of our study show that these factors have a greater presence in female, mixed breed, uncastrated, and older dogs, confirming the data in the veterinary literature. In the present study, we found different histological types of malignant breast tumors with mutations in the BRCA1 gene, as other authors have reported. However, we also found the mutation 3761C>T, which, to the best of our knowledge, has not been reported in the literature. This shows the need for studies in veterinary medicine that assess mutations in the BRCA1 gene and the most common histological types. In conclusion, SNPs in the BRCA1 gene cause genetic instability, resulting in additional mutations that lead to the development of breast tumors. They are point mutations that affect transcription, resulting in truncated proteins. These proteins may have a loss of function, leading to carcinogenesis.Keywords: polymorphism, BRCA1 gene, cancer, dogs.Descritores:polimorfismo, gene BRCA1, câncer, cães. Título: Presença da mutação do gene BRCA1 em cadelas com tumores mamários malignos.

Report of cerebellar hypoplasia in three calves

Hereditary or acquired cerebellar hypoplasia (CH) is commonly diagnosed in Holstein, Guernsey, Shorthorn and Jersey cattle. Bovine viral diarrhea (BVD) has been associated to acquired CH due to viral infection during the second trimester of pregnancy. Stricken calf usually shows ataxia, hypermetria, opisthotonus, intentional tremor and wide-based stance when in standing position. Three newborn calves were referred to the FCAV/Unesp Veterinary Teaching Hospital because of neurological distress. The clinical presentation, similar in all cases, indicated CH. Two weeks later, clinical signs did not improve and euthanasia was performed. Macroscopic examination revealed a gelatinous serosanguineous fluid over the brain surface and within the cervical spinal canal. Histologically the cerebellum had disorganization of the internal granular layer and moderate disappearance of Purkinje cells. The observed clinical signs and nervous tissue lesions were consistent with congenital cerebellar syndrome, possibly associated to viral infection during fetal development. Despite CH has been assumed to be related to BVD, blue tongue and Akabane viruses, only the BVD etiology has been already identified in Brazil

Cerebral Cavernous Hemangioma in a Dog

Background: Cerebral cavernous hemangioma is a rare neoplasm of vascular origin in the brain, characterized by abnormally dilated vascular channels surrounded by endothelium without muscle or elastic fibers. Presumptive diagnosis is performed by magnetic resonance or computed tomography (CT) scanning and can be confirmed by histopathology. The prognosis of intracranial cavernous hemangioma is poor, with progression of clinical signs culminating in spontaneous death or euthanasia. The purpose of this paper is to report a case of cerebral cavernous hemangioma in a dog, presenting the clinical findings, tomographic changes, and pathological findings.Case: This case involved a 2-year-old medium sized mixed breed female dog presenting with apathy, hyporexia, ataxia, bradycardia, dyspnea, and seizure episodes for three days. Hemogram and serum biochemistry of renal and hepatic function and urinalysis did not reveal any visible changes. CT scanning was also performed. The scans revealed a hyperdense nodule of 15.9 x 14 mm, with well defined borders, and a hypodense halo without post-contrast enhancement and mass effect in the right parietal lobe was observed in both transverse and coronal sections. Based on the image presented in the CT scans, the nodule was defined as a hemorrhagic brain lesion. The animal died after a seizure. The right telencephalon was subjected to necropsy, which revealed a reddish-black wel-defined nodule 1.7 cm in diameter extending from the height of the piriform lobe to the olfactory trine at the groove level and extending towards the lateral ventricle, with slight compression and deformation of the thalamus but no other macroscopic alterations in the other organs. The histopathology indicated that this nodular area in the encephalus contained moderate, well-delimited but unencapsulated cellularity, composed of large vascular spaces paved with endothelial cells filled with erythrocytes, some containing eosinophilic fibrillar material (fibrin) and others with organized thrombus containing occasional neutrophil aggregates. The endothelial cells had cytoplasm with indistinct borders, elongated nuclei, scanty crust-like chromatin, and cellular pleomorphism ranging from discrete to moderate, without mitotic figures.Discussion: The histological findings characterized the morphological changes in the brain as cavernous hemangioma, and the growth and compression of this neoplasm were considered the cause of the clinical signs of this dog. The main complaint was seizures, although ataxia and lethargy were also noted. These clinical signs are often related to changes in the anterior brain and brainstem. The literature does not list computed tomography as a complementary diagnostic method in cases of cerebral cavernous hemangioma in dogs, but CT scanning was useful in confirming cerebral hemorrhage. The main differential diagnosis for cerebral cavernous hemangioma would be a hamartoma, but what differentiates them histologically is the presence of normal interstices between the blood vessels, since no intervening neural tissue occurs in the case of cerebral hemangioma. Therefore, even in the absence of immunohistochemistry to more confidently confirm a cavernous hemangioma, the clinical signs, CT scans and especially the pathological findings were consistent with a case of cerebral cavernous hemangioma, a benign neoplasm with a poor prognosis due to the severe neurological changes it causes and its difficult treatment

Feline platynosomiasis: analysis of the association of infection levels with pathological and biochemical findings

Abstract Platynosomiasis is a common feline hepatic disease caused by Platynosomum fastosum (Trematoda - Dicrocoelidae), which is also known as ‘lizard poisoning’. Most reports of feline platynosomiasis show that this disease is sporadic and manifests with uncommon lesions; its pathogenicity is still not well understood. This study aimed to describe liver injuries and enzymatic changes associated with natural P. fastosum infection in 47 stray cats in an endemic area. Overall, 38.3% (18/47) of cats were parasitized, and 2,358 flukes (P. fastosum) were collected (131 – mean intensity of parasitism; 50.2 – mean abundance). The alanine transaminase (ALT) measure was significantly higher in parasitized animals, while alkaline phosphatase (ALP) showed no statistical difference between parasitized and non-parasitized animals. In infected animals, gross pathological lesions and microscopic liver injuries ranged from mild to severe, and were similar to those in previous descriptions of feline platynosomiasis. Nonetheless, the intensity of parasitism was not related to the severity of macroscopic or microscopic hepatic injuries. However, feline platynosomiasis should be considered in the differential diagnosis of feline liver disorders, as well as, in any program of helminth control, even if no clinical abnormalities are present