• Korean Journal of Veterinary Service
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• v.29 no.4
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• pp.497-501
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• 2006

Acute hemorrhagic enteritis was diagnosed in a seven-month-old male Shitzu dog dying of blood stained diarrhea and vomiting. Clinical findings were anorexia, dullness and sudden death after massive bloody diarrhea. At necropsy, main lesion was the hemorrhage in small intestine, mainly duodenum and jejunum. Microscopically, Gram positive long bacilli were massively detected on the mucose epithelial cells and necrotic debris of small intestine. Coagulative necrosis of epithelial cells and thrombosis of small intestine were also identified. However, there was no lesion of crypt epithelium. Mineral infiltration in both gastric mucosa and renal tubules was detected and proliferation of fibrous tissue was also shown in corticomedullary regions. In bacterial examination, C perfringens was isolated in anaerobic culture and it was confirmed to type A by multiplex PCR. Therefore, the dog was diagnosed as C perfringens type A associated enteritis with uremia.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1496-1511
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• 2015

The study aimed to compare the necrotic enteritis (NE)-induced transcriptome differences between the spleens of Marek's disease resistant chicken line 6.3 and susceptible line 7.2 co-infected with Eimeria maxima/Clostridium perfringens using RNA-Seq. Total RNA from the spleens of two chicken lines were used to make libraries, generating 42,736,296 and 42,617,720 usable reads, which were assembled into groups of 29,897 and 29,833 mRNA genes, respectively. The transcriptome changes were investigated using the differentially expressed genes (DEGs) package, which indicated 3,255, 2,468 and 2,234 DEGs of line 6.3, line 7.2, and comparison between two lines, respectively (fold change ${\geq}2$, p<0.01). The transcription levels of 14 genes identified were further examined using qRT-PCR. The results of qRT-PCR were consistent with the RNA-seq data. All of the DEGs were analysed using gene ontology terms, the Kyoto Encyclopedia of Genes and Genomes (KEGG) database and the DEGs in each term were found to be more highly expressed in line 6.3 than in line 7.2. RNA-seq analysis indicated 139 immune related genes, 44 CD molecular genes and 150 cytokines genes which were differentially expressed among chicken lines 6.3 and 7.2 (fold change ${\geq}2$, p<0.01). Novel mRNA analysis indicated 15,518 novel genes, for which the expression was shown to be higher in line 6.3 than in line 7.2 including some immune-related targets. These findings will help to understand host-pathogen interaction in the spleen and elucidate the mechanism of host genetic control of NE, and provide basis for future studies that can lead to the development of marker-based selection of highly disease-resistant chickens.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.10
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• pp.1478-1485
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• 2017

Objective: The effects of vaccinating 18-day-old chicken embryos with the combination of recombinant Eimeria profilin plus Clostridium perfringens (C. perfringens) NetB proteins mixed in the Montanide IMS adjuvant on the chicken immune response to necrotic enteritis (NE) were investigated using an Eimeria maxima (E. maxima)/C. perfringens co-infection NE disease model that we previously developed. Methods: Eighteen-day-old broiler embryos were injected with $100{\mu}L$ of phosphate-buffered saline, profilin, profilin plus necrotic enteritis B-like (NetB), profilin plus NetB/Montanide adjuvant (IMS 106), and profilin plus Net-B/Montanide adjuvant (IMS 101). After post-hatch birds were challenged with our NE experimental disease model, body weights, intestinal lesions, serum antibody levels to NetB, and proinflammatory cytokine and chemokine mRNA levels in intestinal intraepithelial lymphocytes were measured. Results: Chickens in ovo vaccinated with recombinant profilin plus NetB proteins/IMS106 and recombinant profilin plus NetB proteins/IMS101 showed significantly increased body weight gains and reduced gut damages compared with the profilin-only group, respectively. Greater antibody response to NetB toxin were observed in the profilin plus NetB/IMS 106, and profilin plus NetB/IMS 101 groups compared with the other three vaccine/adjuvant groups. Finally, diminished levels of transcripts encoding for proinflammatory cytokines such as lipopolysaccharide-induced tumor necrosis $factor-{\alpha}$ factor, tumor necrosis factor superfamily 15, and interleukin-8 were observed in the intestinal lymphocytes of chickens in ovo injected with profilin plus NetB toxin in combination with IMS 106, and profilin plus NetB toxin in combination with IMS 101 compared with profilin protein alone bird. Conclusion: These results suggest that the Montanide IMS adjuvants potentiate host immunity to experimentally-induced avian NE when administered in ovo in conjunction with the profilin and NetB proteins, and may reduce disease pathology by attenuating the expression of proinflammatory cytokines and chemokines implicated in disease pathogenesis.

• Korean Journal of Veterinary Research
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• v.33 no.3
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• pp.419-427
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• 1993

Thirteen strains of Clostridium perfringens were isolated from the pigs with hemorrhagic enteritis. The characteristics of the outbreaks, clinical signs and lesions were examined. The biochemical properties, type of toxin and susceptibility to antimicrobial agents of the isolates were investigated. The results could be summarized as follows ; 1. Almost of the pigs affected with hemorrhagic enteritis, 17 cases examined from 1989 to 1992, were piglets less than 7 day old. 2. The average mortality rate of piglet less than 7 day old affected with hemorragic and necrotic enteritis was 48.5%. 3. The clinical signs of pigs with hemorrhagic enteritis were depression, hemorrhagic diarrhea, anemia and dehydration. Necropsy of the infected pigs showed typical hemorrhage of upper intestine and necrosis of mucosal membrane. 4. The characteristic biochemical properties of the isolates were 2-band hemolysis, positive reaction of reverse CAMP test and formation of LV precipitate in egg yolk medium. 5. The toxin type of the 13 isolates, investigated by mouse inoculation test, was all type C strains of Clostridium perfringens. 6. In susceptibility test to antimicrobial agents, 13 isolates of Clostridium perfringens were highly sensitive to ampicillin, enrofloxacin(Baytril), cephalothin, penicillin and trimethoprim-sulfamethoxazole.

• Journal of Veterinary Clinics
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• v.15 no.1
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• pp.180-183
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• 1998

A 5 year-old male Throughbred horse housed in Korean Racing Association died after having enteritis and respiratory disorder for about 10 days. At necropsyi the left and right caudal lung lobes were reddenedi swolleni and contained numerous well-defined sublobular consolidated foci. The large intestine was diffusely reddened and covered with pseudomembranous exudates. Microscopicallyi multifocal areas of necrosis with mild to moderate infiltration of neutrophilsi macrophages and Iymphocytes as well as hemorrhage and edema were noted in the lung. The fungi having conidiophorei dome-shaped vesicles phialides and conidia which are characteristic of Apergillosis fumigatus was isolated from the lung. The colonic mucosa was transmurally necrotic and severe congestions edema and thrombi were observed in the submucosa. The embolic mycotic pneumonia present in this case Probably occured secondary to antibiotic therapy given for treatment of enteritis.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.829-837
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• 1996

Clostridium perfringens has been identified as a causative organism in necrotic enteritis in chicken. The bacterium has been classified into five toxigenic types (A through E) based on the pattern of the production of major lethal toxins. Seroneutralization with mice or guinea pigs usually has been used to type the organism. Of the types, types A and C of the bacterium had been recognized as the major pathogenic types in chicken. In this experiment, we isolated nine field strains of C perfringens from chicken showing necrotic enteritis in clinical symptoms and pathologic findings and identified by biochemical tests. In order to type the organism, a multiplex polymerase chain reaction (PCR) was used with primers on major lethal toxin genes instead of seroneutralization. Amplification of only a toxin gene with the PCR suggested that the disease in chicken was due to type A of C perfringens in Korea. Furthermore, the PCR method can be replaced with seroneutralization to type C perfringens in future.

• Korean Journal of Veterinary Research
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• v.47 no.4
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• pp.437-441
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• 2007

The cases of feline panleukopenia virus (FPLV) infection were diagnosed in three imported cats. All cats died within one week after mild emaciation, depression and anorexia. One cat showed yellowish watery diarrhea. At necropsy, all cats had segmental hemorrhage on the serosa and mucosa of the small intestine. Histopathologically, severe diffuse necro-hemorrhagic enteritis was observed in small intestine especially in jejunum and ileum. The crypts of Lieberkuhn were dilated and contained necrotic epithelia. Severely damaged epithelia of crypts were transformed into bizarre shapes. Multifocal lympholysis and lymphoid depletion were found in Peyer's patches and other lymphoid tissues. Direct fluorescent antibody (FA) test revealed the characteristic FPLV antigen in the cytoplasms of crypt epithelial cells. Based on the clinical signs, characteristic pathologic findings and FA test, these cases were diagnosed as FPLV infection. In our best knowledge, this study is the first case report for FPLV infection in imported cats in Korea.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.7
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• pp.1037-1047
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• 2017

Objective: Despite an increasing number of investigations into the pathophysiology of necrotic enteritis (NE) disease, etiology of NE-associated diseases, and gene expression profiling of NE-affected tissues, the microRNA (miRNA) profiles of NE-affected poultry have been poorly studied. The aim of this study was to induce NE disease in the genetically disparate Fayoumi chicken lines, and to perform non-coding RNA sequencing in the intestinal mucosal layer. Methods: NE disease was induced in the Fayoumi chicken lines (M5.1 and M15.2), and non-coding RNA sequencing was performed in the intestinal mucosal layer of both NE-affected and uninfected chickens to examine the differential expression of miRNAs. Next, quantitative real-time polymerase chain reaction (real-time qPCR) was performed to further examine four miRNAs that showed the highest fold differences. Finally, bioinformatics analyses were performed to examine the four miRNAs target genes involvement in the signaling pathways, and to examine their interaction. Results: According to non-coding RNA sequencing, total 50 upregulated miRNAs and 26 downregulated miRNAs were detected in the NE-induced M5.1 chickens. While 32 upregulated miRNAs and 11 downregulated miRNAs were detected in the NE-induced M15.2 chickens. Results of real-time qPCR analysis on the four miRNAs (gga-miR-9-5p, gga-miR-20b-5p, ggamiR-196-5p, and gga-let-7d) were mostly correlated with the results of RNAseq. Overall, ggamiR-20b-5p was significantly downregulated in the NE-induced M5.1 chickens and this was associated with the upregulation of its top-ranking target gene, mitogen-activated protein kinase, kinase 2. Further bioinformatics analyses revealed that 45 of the gene targets of gga-miR-20b-5p were involved in signal transduction and immune system-related pathways, and 35 of these targets were predicted to interact with each other. Conclusion: Our study is a novel report of miRNA expression in Fayoumi chickens, and could be very useful in understanding the role of differentially expressed miRNAs in a NE disease model.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.4
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• pp.541-551
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• 2012

The effects of dietary additives and holding time on resistance and resilience of broiler chickens to Clostridium perfringens challenge were investigated by offering four dietary treatments. These were a negative control (basal), a positive control (Zn-bacitracin) and two dietary additives, mannanoligosaccharides (MOS), and acidifier. Two holding times included (a) immediate access to feed and water post hatch (FED) and (b) access to both feed and water 48 h post hatch (HELD). Chicks fed Zn-bacitracin had no intestinal lesions attributed to necrotic enteritis (NE), whereas chicks fed both MOS or acidifier showed signs of NE related lesions. All dietary treatments were effective in reducing the numbers of C. perfringens in the ileum post challenge. The FED chicks had heavier body weight and numerically lower mortality. The FED chicks also showed stronger immune responses to NE challenge, showing enhanced (p<0.05) proliferation of T-cells. Early feeding of the MOS supplemented diet increased (p<0.05) IL-6 production. The relative bursa weight of the FED chicks was heavier at d 21 (p<0.05). All the additives increased the relative spleen weight of the HELD chicks at d 14 (p<0.05). The FED chicks had increased villus height and reduced crypt depth, and hence an increased villus/crypt ratio, especially in the jejunum at d 14 (p<0.05). The same was true for the HELD chicks given dietary additives (p<0.05). It may be concluded that the chicks with early access to dietary additives showed enhanced immune response and gut development, under C. perfringens challenge. The findings of this study shed light on managerial and nutritional strategies that could be used to prevent NE in the broiler industry without the use of in-feed antibiotics.

• Korean Journal of Poultry Science
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• v.44 no.3
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• pp.211-223
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• 2017

Transforming growth factor beta ($TGF-{\beta}$) signaling pathways are involved in the regulation of proliferation, differentiation, immunity, survival, and apoptosis of many cells. The aim of this study was to investigate the differential expression of $TGF-{\beta}$-related genes, and their interactions and regulators in the spleen of two genetically disparate chicken lines (Marek's disease resistant line 6.3 and Marek's disease-susceptible line 7.2) induced with necrotic enteritis (NE) by Eimeria maxima and Clostridium perfringens infection. By using high-throughput RNA-sequencing, we investigated 76 $TGF-{\beta}$-related genes that were significantly and differentially expressed in the spleens of the chickens. Approximately 20 $TGF-{\beta}$ pathway genes were further verified by qRT-PCR, and the results were consistent with our RNA sequencing data. All 76 identified genes were analyzed through Gene Ontology and mapped onto the KEGG chicken $TGF-{\beta}$ pathway. Our results demonstrated that several key genes, including $TGF-{\beta}$1-3, bone morphogenetic proteins (BMP)1-7, inhibitor of differentiation (ID) proteins ID1-3, SMAD1-9, and Jun, showed a markedly differential expression between the two chicken lines, relative to their respective controls. We then further predicted 24 known miRNAs that targeted BMP7 mRNA from 139 known miRNAs in the two chicken lines. Among these, six miRNAs were measured by qRT-PCR. In conclusion, this study is the first to analyze most of the genes, interactions, and regulators of the $TGF-{\beta}$ pathway in the innate immune responses of NE afflicted chickens.