We conducted diagnostic investigations to analyze the causes of abortions (46 cases, 65.7%), deaths (22 cases, 31.4%) and muscular lesions (2 cases, 2.9%) occurred after foot and mouth disease (FMD) vaccination in livestock farms in Korea. Bacterial culture, enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) were performed to detect the causative agents of abortion in bovine and caprine. The diagnostic results showed that 36 (51.4%) cases, referring as “Identified”, were occurred by influence of underlying disease including bovine viral diarrhea (12 cases, 17.1%), neosporosis (7 cases, 10.0%), septicemic colibacillosis (5 cases, 7.1%), Q fever (4 cases, 5.7%) and other abnormal conditions (8 cases, 11.4%) not by vaccination. Other 2 (3.0%) cases were suspected to be vaccine-associated adverse reaction on the basis of pathological findings (shock lung, oil-component-induced granuloma) and clinical symptoms (dyspnea with pulmonary edema). The other 32 (45.7%) cases were determined “Unknown” because any pathogens and pathological changes were not identified. However, many of the “Unknown” cases were presumptive to be the vaccine-related adverse reaction based on epidemiological investigation, especially, the cases which showed the clinical signs within 2 days after the vaccination. It is important to conduct pathological, microbilogical and epidemiological investigation to diagnose whether the cases are from vaccine-associated adverse reaction or not.
A 19-year-old male Siberian tiger was presented with inappetence and paralysis of hind limbs. In a computed tomography (CT) scan, intervertebral disk disease at L3–L4 was detected. Cardiac arrest occurred during the surgery. At autopsy, myxomatous mitral valve degeneration (MMVD) and eccentric hypertrophy of the left heart were noted. The diagnosis was congestive heart failure caused by MMVD. Microscopically, myocardial and pulmonary fibrosis were observed in addition to the disintegration of the fibrosa layer and accumulation of glycosaminoglycans and proteoglycans in the spongiosa layer of the mitral valve. This is the first case of congestive heart failure with MMVD in a Siberian tiger.
We report a massive outbreak of human Q fever cases, which occurred at totally 11 humans. The occurrence was related to a goat farm where Coxiella burnetii infection was diagnosed from goat tissues and environmental specimens. From January of 2018, continuous abortions from 6 goats occurred. Laboratory tests from 77 goat specimens for C. burnetii showed that 54 (70.1%) and 63 (81.8%) goats were positive by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. The infection was also confirmed from the farmer, his wife and 9 persons from all 16 veterinary officials who had visited the farm for protective measures and preparing goat specimens for laboratory tests. The farm was found to be extensively contaminated with C. burnetii from the examination to the environmental specimens and epidemiological inspections, which might be the main source of C. burnetii infection to humans. The extensive contamination to the farm was derived from the uncareful handling of postpartum animal tissues or discharges by the farm owner. This report will contribute to the establishment of educational system on the biosecurity to novice farmers.
α-Viniferin (AVF), a trimer of resveratrol, is known to have an anti-inflammatory effect via inhibition of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). It has been reported that up-regulated COX-2 and iNOS are expressed in colon cancer tissues of humans and rodents as well as pre-neoplastic aberrant crypt foci (ACF) of rodents. In this study, chemopreventive effects of AVF were assessed in Caco-2 cells as well as azoxymethane (AOM)-induced colorectal tumorigenesis in mice. Anti-tumor effect of AVF with regards to apoptotic induction was assessed by TUNEL and caspase-3 expression in human colon cancer Caco-2 cells. For development of ACF, AOM was administered with to mice intraperitoneally at a dose of 10 mg/kg once a week for 3 weeks. To induce colitis-related colon cancer, mice were administered a single dose of AOM (10 mg/kg) and 2% dextran sodium sulfate in drinking water. Mice treated with 0.05 and/or 0.1 mg of AVF by gavage showed significantly reduced development of ACF and colorectal tumors. Immunofluorescence detection in Caco-2 cells showed reduced COX-2 and iNOS expression, whereas cleavage of caspase-3 and apoptotic cell numbers increased upon AVF treatment. Immunostaining showed reduced expression levels of COX-2 and iNOS expression along with increased cleaved caspase-3 expression increased upon AVF treatment. These results suggest that AVF has chemopreventive effects on colorectal cancer via anti-inflammatory potential and pro-apoptotic activity.
Pulmonary fibrosis (PF) is defined as a gradual, interstitial fibrous disease of the lung parenchyma. PF causes collapse of the lung lobe, which can lead to lung lobe torsion of the other side. A 14-year-old male Pekinese dog was referred to the Veterinary Medical Center, Chungbuk National University. The chief complaint was acute dyspnea. The dog had a history of chronic cough, which lasted for 18 months, and the cough had recently deteriorated. Tachypnea was observed on physical examination. As a result of thoracic radiographs, ultrasonography, and computed tomography, lung lobe torsion and collapse were diagnosed. A postmortem examination revealed lung lobe torsion of the left cranial lobe and carnification of the right cranial lobe. Histologically, severe and diffuse interstitial fibrosis with distortion of alveolar architecture and severe congestion and/or hemorrhage were observed in the right cranial lobe and left cranial lobe, respectively. Although the cause of pulmonary fibrosis was undetermined, this case showed a typical lung lobe torsion caused by pulmonary fibrosis.
Immunotherapeutic approaches using agonist antibodies or fusion proteins of immunomodulatory molecules significantly inhibit tumor growth and boost cell-mediated immunity. We isolated mRNA from previously reported 1D4 hybridoma cells and amplified the variable regions of the heavy chain (VH) and light chain (VL) genes using reverse-transcriptase polymerase chain reaction. Using a linker, the amplified sequences for the heavy and light chains were each connected to the sequence for a single polypeptide chain that was designed to be expressed. VL and VH fragments were cloned into the pOptiVEC-TOPO vector containing the human CH2-CH3 fragment. Then, 293T cells were transfected with the 1D4 single-chain Fv-Fc (scFv-Fc) constructs. A549 cells were used for presentation of the 1D4 antigen. Flow cytometry was performed for analysis of the secreted 1D4 scFv-Fc constructs. The DNA sequence of 1D4 scFv-Fc was obtained. The 1D4 scFv-Fc constructs were expressed by the transfected 293T cells and secreted into the culture medium. The immunoreactivity of the secreted scFv-Fc construct was lower than that of the murine 1D4 antibody for A549 cells. A 1D4 scFv-Fc construct for immunotherapy was developed.
Canine heartworm disease is typically characterized by the presence of heartworms in the pulmonary artery and, to a lesser degree, the right ventricle. Systemic arterial dirofilariasis is an unusual manifestation of heartworm disease of dogs that results from aberrant migration of Dirofilaria immitis into the peripheral arterial circulation. A three-year-old Jindo dog was referred to the Veterinary Medical Center because of hindlimb lameness, paresthesia of hindlimbs, and interdigital ischemic necrosis. Angiography and ultrasonography were performed, resulting in diagnosis of heartworm infestation. In necropsy, entangled heartworms were present throughout the body, including lungs, kidney, small intestinal lumen, and arteries, in addition to the right ventricle and pulmonary artery. Histologically, lung showed adult heart worms in vessels, organized thrombotic vessels, and interstitial pneumonia featuring type II pneumocyte hyperplasia, thickened alveolar wall, and compressed alveolar spaces. Chronic parenchymal lesions, such as focal or diffuse necrosis, fibrosis, and dystrophic calcification were remarkable in liver and kidney. The current case showed a typical canine systemic heartworm disease caused by aberrant migration of Dirofilaria immitis.
A six-year-old male Pekinese dog was referred to the Veterinary Medical Center, Chungbuk National University because of intermittent vomiting, anorexia, alopecia, and pruritus. Generalized alopecia, pigmentation, and papular erythema in the skin were observed on physical examination. Hematological studies indicated a severe pancytopenia and hypoalbuminemia. A hormone analysis indicated a hyperestrogenemia. A circumscribed mass, measuring approximately 3-4 cm in diameter, was observed on abdominal radiographs. Grossly, the cryptorchid testis was enlarged by a firm and white spherical mass, measuring approximately 3 cm in diameter, which, on its cut surface, was creamy white with hemorrhage. The normal scrotal testis was markedly atrophic and soft. Histologically, the intra-abdominal cryptorchid testis contained abundant fibrous tissue stroma and the stromal tissues were arranged in a tubular pattern in which the neoplastic cells tended to palisade. The cells had poor and pale eosinophilic cytoplasm streaming into the center of the tubules. The nuclei were round to oval and tended to be in a basilar location. Regions of hyperplasia and marked squamous metaplasia were observed in many areas of the prostate. Based on the histological findings, we were able to identify these masses as Sertoli cell tumors, and made a final diagnosis as Sertoli cell tumors through immunohistochemistry methods using inhibin-α, vimentin, and neuron-specific enolase.
Cutaneous vasculitis is an inflammatory necrotizing disease of the blood vessel walls. In most cases, the etiology is due to immune-complex disease from type III hypersensitivity. Clinical signs include swelling, erythema, purpura, erythematous plaques, ulceration, and necrosis. Clinical lesions may be localized to distal extremities or areas of less extensive collateral circulation. In this case, history, physical examination, laboratory findings, cytology, histopathologic examination, and immunohistochemistry might aid in differentiation of a clinically similar disease. Consequently, this case was diagnosed as cutaneous vasculitis and treated with ointment containing corticosteroid and antibiotics over a period of two weeks. After treatment, all of the clinical lesions had disappeared.
Chronic inflammatory diseases such as Crohn′s disease and ulcerative colitis are associated with increased risk of colon adenocarcinoma. Apoptic induction of colon cancer cells by cytokines and death receptors is an important anti-cancer therapy. We observed that co-administration of TNFα and IFNγ in human colon cancer cell line, HCT116, resulted in cell death and expression of IL-32. Cleavage forms of caspase-3, caspase-9, and PARP were increased in TNFα / IFNγ-treated HCT116. mRNA expression of death receptors, including TNFR1 and Fas were not changed and NO generation was not induced by combination of TNFα and IFNγ. However, mRNA expression of IL-32α, β, and γ was increased in TNFα / IFNγ-treated HCT116. To determine the effect of IL-32 in HCT116 cell apoptosis by TNFα / IFNγ stimulation, IL-32 siRNA-transfected HCT116 cells were cultured with TNFα / IFNγ and cell proliferation was measured. IL-32 siRNA induced slight recovery of cell viability of TNFα / IFNγ-stimulated HCT116. These results suggest that IL-32 is not directly related to apoptosis of HCT116 by TNFα / IFNγ stimulation. However, IL-32 expression by TNFα or TNFα / IFNγ in a colon cancer cell line is very interesting because of the unknown effect of IL-32 in colon cancer. Our study will contribute to development of studies for IL-32 function in human colon cancer and anti-cancer therapies using cytokines.
Histologic and immunohistochemical features of canine skin tumours originated from epithelial stem cells were described. The tumors were trichogenic trichoblastoma, trichoepithelioma, sebaceous adenoma, hepatoid gland adenoma and epithelioma, and basal cell tumour. All of the tumors expressed both AE1/AE3 and cytokeratin 5/8, whereas cytokeratin 7 and vimentin not at all. Neuron-specific-enolase was strongly positive in follicular tumours, whereas slightly in basal cell tumour. Moreover, chromogranin A was expressed slightly only in trichogenic trichoblastoma and trichoepithelioma. Since the morphological similarities between these tumors, immunohistochemistry is one of useful tools for differential diagnosis. From the immunohistochemistry data, we conclude that cytokeratins, vimentin and NSE play a main role in the evaluation of canine skin tumours originated from epithelial skin stem cells histogenesis.
Immunohistochemical localization of vimentin, glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), neuron-specific enolase (NSE) were performed to confirm the canine schwannoma diagnosed histologically. Biopsy samples from skin masses occurred on a 6 year old female dog and a 13 year old female dog were used for immunohistochemical analysis. Histologically, the tumor tissues showed the characteristics of schwannoma mainly composed of predominantly fusiform or spindle cells. The cells are arranged in small interwoven bundles with intervening fibrous and mucinous stroma that imitate a herringbone pattern. The cell nuclei have an elongated, wavy, buckled appearnce and closely bunched together. In high magnification, mitotic figure, nuclear hyperchromatism, pleomorphism and poorly defined cytoplasms were noted. the vimentin was highly expressed in the cytoplasm of all neoplastic cells. Positive reactions against NSE and GFAP, widely used to suggest a neural origin of cell types, are observed in cytoplasms of tumor cells. These results can be used as a reference data for definitive diagnosis of canine schwannoma.
A 13-year-old, male mixed breed dog was presented to Veterinary Medical Center, Chungbuk National University for diagnosis and treatment of enlarged mass in right inguinal region. On hematological test, monocytes and neutrophils were slight increased, but other hematological values were normal. On radiological finding, opacity and size of mass in right inguinal region were increased. The mass and testis were removed surgically. Removed mass was oval-shpae and bulky with 5.6×7.6×5.2 cm. Histopathological finding of the mass revealed large and round to polyhedral cells and it was diagnosed as malignant seminoma. That was diffuse type seminoma near to malignant tumor without metastasis. During the follow-up for 13 months, patient showed normal contition without recurrence.
Phytic acid (PA) is a naturallu occurring polyphosphorylated carbohydrate that is present in substantial amounts in almost all plants and mammalian cells. Recently PA has received much attention for its role in anticancer activity. We investigated the preventive effect of PA on the formation of colonic aberrant crypt foci (ACF), a preneoplastic lesion, induced by azoxymethane (AOM). After acclimation for one week, six-week old male ICR mice were fed on the AIN-93G purified diet and PA (0.5% or 2% PA in water) for 8 weeks. The animals were treated with azoxymethane (AOM, 10 mg/kg b.w.) three times (0, 1, and 2 weeks) to induce colonic aberrant crypt foci (ACF). After sacrifice, the total numbers of aberrant crypts (AC) and ACF in colonic mucosa were counted after staining with methylene blue. Blood and serum were analyzed with a blood cell differential counter and an automatic serum analyzer. AOM treatment without PA induced the total numbers of 85.7 ± 12.9 and 115.2 ± 19.9, respectively. PA at the dose of 2% AC/colon by PA at the dose of 0.5% were 73.4 ± 12.9 and 115.2 ± 19.9, respectively. PA at the dose of 2% significantly decreased the ACF and AC numbers to 56.5 ± 14.6 and 95.4 ± 17.2, respectively (p<0.01). PA at the doses of 0.5 and 2% decreased the numbers of ACF and AC/colon in a dose-dependent manner. Although some parameters in blood counts and serum chemistry were changed compared with the control, no specific toxicity was found. Theses findings suggest that phytic acid can be a chemopreventive agent for colon carcinogenesis resulting from inhibition of the development of ACF in ICR mice.