1.Evaluation of assays to detect Helicobacter felis infection in cats.
Journal of Biomedical Research 2015;16(4):159-164
Recently, several companies have released H. pylori stool antigen (HpSA) test kits. However, there is little information about the usefulness of HpSA testing for Helicobacter felis, which is the major Helicobacter species in cats. The aim of the present study was to compare diagnostic methods for diagnosis of H. felis with HpSA tests and PCR assay using cat stools or gastric mucosa. Male cats (n=6) were infected with H. felis ATCC 49179 (1.0 x 10(9) CFU /cat) by intragastric inoculation two times at 3-day intervals, and stool specimens of cats were collected 1, 3, 5, 7, 14, and 21 days after infection for HpSA testing and H. felis-specific PCR. For the results, sensitivities of the HpSA test and PCR analysis were 50.0% and 83.3% respectively. Cats were sacrificed 21 days after H. felis inoculation, and gastric tissues were homogenized. All gastric biopsy specimens were positive based on a rapid urease test (RUT) (6/6, 100%) and PCR (6/6, 100%). Based on these results, the HpSA kit is useful and effective for monitoring H. felis infection using stool specimens. If an HpSA test could be made with H. felis antibodies in the future, its sensitivity could be increased further. Further, PCR assay could be successfully used to detect H. felis in stools. Application of this HpSA kit and PCR assay can be utilized as a non-invasive strategy to identify H. felis in cats.
Animals
;
Antibodies
;
Biopsy
;
Cats*
;
Diagnosis
;
Felis
;
Gastric Mucosa
;
Helicobacter felis*
;
Helicobacter*
;
Humans
;
Male
;
Natural Resources
;
Polymerase Chain Reaction
;
Urease
2.Molecular identification of Mycoplasma cynos from laboratory beagle dogs with respiratory disease.
Laboratory Animal Research 2012;28(1):61-66
In this study, we examined a colony of 20 beagle dogs in a laboratory animal facility. Mycoplasma was detected by consensus PCR assay in 1 dog with respiratory and constitutional symptoms. None of the other dogs were affected. The dog was euthanized and necropsied. In postmortem examinations, gray or plum-colored gross lesions were found on the lung, most commonly in the apical and cardiac lobes. Some lesions showed clear demarcation and consolidation. Microscopic examination showed peribronchiolar lymphoid hyperplasia and interstitial thickening, lesions pathognomonic for mycoplasma pneumonia. To identify canine Mycoplasma species, we used species-specific PCR reactions for M. arginini, M. canis, M. cynos, M. edwardii, M. felis, M. gateae, M. maculosum, M. molare, M. opalescens, M. spumans, Mycoplasma sp. HRC 689, and M. collis. As the result, we identified Mycoplasma cynos by amplification of DNA extracted from lung tissue of the laboratory beagle dog with respiratory disease.
Animals
;
Animals, Laboratory
;
Autopsy
;
Cats
;
Consensus
;
DNA
;
Dogs
;
Felis
;
Hyperplasia
;
Lung
;
Molar
;
Mycoplasma
;
Pneumonia, Mycoplasma
;
Polymerase Chain Reaction
3.Postpartum Transmission as a Major Route of Mother-to-Child Helicobacter felis Infection.
Sunhwa HONG ; Hyun A LEE ; Youngho KIM ; Okjin KIM
Laboratory Animal Research 2010;26(3):319-321
In this study we investigated maternal Helicobacter felis (H. felis) infection status to determine the potential of maternal transmission. Pregnant Beagle dogs were infected experimentally with H. felis. Following the experimental design, the stools of the mother and litters were isolated and assessed for transmission of H. felis at parturition day, 1-week old age and 6-week old age respectively. Polymerase chain reaction (PCR) was used to examine the presence of transmitted H. felis. All litters showed no transmission of H. felis at parturition day. However, they revealed 14.3% and 100% at 1-week old age and 6-week old age respectively by PCR. These results suggested that vertical infection during prenatal period or delivery procedure is unlikely as a route of mother-to-child H. felis infection. It might be acquired H. felis through breast-feeding, contaminating saliva and fecal-oral during co-habitat.
Animals
;
Cats
;
Dogs
;
Felis
;
Helicobacter
;
Helicobacter felis
;
Humans
;
Mothers
;
Parturition
;
Polymerase Chain Reaction
;
Postpartum Period
;
Research Design
;
Saliva
4.Postpartum Transmission as a Major Route of Mother-to-Child Helicobacter felis Infection.
Sunhwa HONG ; Hyun A LEE ; Youngho KIM ; Okjin KIM
Laboratory Animal Research 2010;26(3):319-321
In this study we investigated maternal Helicobacter felis (H. felis) infection status to determine the potential of maternal transmission. Pregnant Beagle dogs were infected experimentally with H. felis. Following the experimental design, the stools of the mother and litters were isolated and assessed for transmission of H. felis at parturition day, 1-week old age and 6-week old age respectively. Polymerase chain reaction (PCR) was used to examine the presence of transmitted H. felis. All litters showed no transmission of H. felis at parturition day. However, they revealed 14.3% and 100% at 1-week old age and 6-week old age respectively by PCR. These results suggested that vertical infection during prenatal period or delivery procedure is unlikely as a route of mother-to-child H. felis infection. It might be acquired H. felis through breast-feeding, contaminating saliva and fecal-oral during co-habitat.
Animals
;
Cats
;
Dogs
;
Felis
;
Helicobacter
;
Helicobacter felis
;
Humans
;
Mothers
;
Parturition
;
Polymerase Chain Reaction
;
Postpartum Period
;
Research Design
;
Saliva
5.Uterine adenocarcinoma with feline leukemia virus infection.
Sung Jin CHO ; Hyun A LEE ; Sunhwa HONG ; Okjin KIM
Laboratory Animal Research 2011;27(4):347-351
Feline endometrial adenocarcinomas are uncommon malignant neoplasms that have been poorly characterized to date. In this study, we describe a uterine adenocarcinoma in a Persian cat with feline leukemia virus infection. At the time of presentation, the cat, a female Persian chinchilla, was 2 years old. The cat underwent surgical ovariohystectomy. A cross-section of the uterine wall revealed a thickened uterine horn. The cat tested positive for feline leukemia virus as detected by polymerase chain reaction. Histopathological examination revealed uterine adenocarcinoma that had metastasized to the omentum, resulting in thickening and the formation of inflammatory lesions. Based on the histopathological findings, this case was diagnosed as a uterine adenocarcinoma with abdominal metastasis. To the best of our knowledge, this is the first report of a uterine adenocarcinoma with feline leukemia virus infection.
Adenocarcinoma
;
Animals
;
Cats
;
Chinchilla
;
Female
;
Horns
;
Humans
;
Leukemia Virus, Feline
;
Leukemia, Feline
;
Neoplasm Metastasis
;
Omentum
;
Polymerase Chain Reaction
;
Uterus
6.An animal model using Eimeria live vaccine and to study coccidiosis protozoa pathogenesis.
Hyun A LEE ; Sunhwa HONG ; Ohmok CHOE ; Okjin KIM
Korean Journal of Veterinary Research 2011;51(3):249-252
Cell culture systems for the protozoan Eimeria are not yet available. The present study was conducted to develop an animal model system by inoculating animals with a live Eimeria vaccine. This study was conducted on 3-day-old chickens (n = 20) pretreated with cyclophosphamide. The chickens were divided into 2 groups: the control group (n = 10) and the inoculated group that received the live Eimeria vaccine (n = 10). During the study period, we compared the clinical signs, changes in body weight, and number of oocysts shed in the feces of the control and inoculated group. This study showed that oocyst shedding was significantly higher in the chickens inoculated with live Eimeria oocysts than in the control chickens. Moreover, body weight gain was lesser in the animals in the inoculated group than in the control animals. Fecal oocyst shedding was observed in the inoculated animals. On the basis of these findings, we suggest that live Eimeria vaccination with cyclophosphamide pretreatment may be used to obtain an effective animal model for studying protozoan infections. This animal study model may eliminate the need for a tedious continuous animal inoculation process every 6 months because the live coccidiosis vaccine contains live oocysts.
Animals
;
Body Weight
;
Cell Culture Techniques
;
Chickens
;
Coccidiosis
;
Cyclophosphamide
;
Eimeria
;
Feces
;
Models, Animal
;
Oocysts
;
Protozoan Infections
;
Vaccination
7.Sensitive and specific identification by polymerase chain reaction of Eimeria tenella and Eimeria maxima, important protozoan pathogens in laboratory avian facilities.
Hyun A LEE ; Sunhwa HONG ; Yungho CHUNG ; Okjin KIM
Laboratory Animal Research 2011;27(3):255-258
Eimeria tenella and Eimeria maxima are important pathogens causing intracellular protozoa infections in laboratory avian animals and are known to affect experimental results obtained from contaminated animals. This study aimed to find a fast, sensitive, and efficient protocol for the molecular identification of E. tenella and E. maxima in experimental samples using chickens as laboratory avian animals. DNA was extracted from fecal samples collected from chickens and polymerase chain reaction (PCR) analysis was employed to detect E. tenella and E. maxima from the extracted DNA. The target nucleic acid fragments were specifically amplified by PCR. Feces secreting E. tenella and E. maxima were detected by a positive PCR reaction. In this study, we were able to successfully detect E. tenella and E. maxima using the molecular diagnostic method of PCR. As such, we recommended PCR for monitoring E. tenella and E. maxima in laboratory avian facilities.
Animals
;
Chickens
;
DNA
;
Eimeria
;
Eimeria tenella
;
Feces
;
Oocysts
;
Pathology, Molecular
;
Polymerase Chain Reaction
8.Spontaneous osteosarcoma of the femur in a non-obese diabetic mouse.
Sunhwa HONG ; Hyun A LEE ; Ohmok CHOE ; Youngho CHUNG ; Okjin KIM
Laboratory Animal Research 2011;27(3):251-254
An abnormal swelling was identified in the distal portion of the right femur in a 1-year-old non-obese diabetic (NOD) mouse. Grossly, a large mass of the distal femur was observed in the right femur. Lesions were poorly marginated, associated with destruction of the cancellous and cortical elements of the bone, and showed ossification within the soft tissue component. Histologically, the tumor was identified as a poorly differentiated sarcoma. Histopathologic examination of the bone masses revealed invasive proliferation of poorly differentiated neoplastic mesenchymal cells forming streams, bundles, and nests, which resulted in destruction of normal bone. Neoplastic cells exhibited random variation in cellular appearance and arrangement, as well as matrix composition and abundance. Haphazard and often intermingling patterns of osteogenic, chondroblastic, lipoblastic, and angiogenic tissues were present. Larger areas of neoplastic bone and hyaline cartilage contained multiple large areas of hemorrhage and necrosis bordered by neoplastic cells. The mass was diagnosed as an osteosarcoma. To our knowledge, this is the first spontaneous osteosarcoma in an NOD mouse.
Animals
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Chondrocytes
;
Durapatite
;
Femur
;
Hemorrhage
;
Hyaline Cartilage
;
Mice
;
Mice, Inbred NOD
;
Necrosis
;
Osteosarcoma
;
Rivers
;
Sarcoma
9.Detection of canine respiratory coronavirus from dogs with respiratory disease.
Sunhwa HONG ; Hak Yong LEE ; Okjin KIM
Journal of Biomedical Research 2015;16(1):19-22
Canine respiratory coronavirus (CRCoV) is commonly associated with canine kennel cough worldwide. Clinically infected dogs present coughing, sneezing, and nasal discharge. Severe infections may progress to pneumonia. Through serological surveys, CRCoV has been identified as a worldwide pathogen found in the respiratory tracts of dogs suffering from mild or severe respiratory disease. In this study, three dogs were obtained from a dog kennel. Over the previous 5 days, the dogs showed coughing, sneezing, and nasal discharge. To detect the etiologic pathogen, we performed multiplex RT-PCR (mRT-PCR) to amplify the genes encoding canine influenza virus matrix protein, canine distemper virus nucleocapsid protein, and CRCoV spike protein. Dot blotting was achieved with a CRCoV-specific probe. Nasal-secreting CRCoV was detected by the 442 bp CRCoV-positive PCR reaction in the nasal swabbing samples from dogs. Further, CRCoV-positive reactions by dot blot hybridization were detected in the nasal swabbing samples from dogs. In conclusion, we detected CRCoV in kenneled dogs with respiratory disease in Korea. Multiplex RT-PCR was able to detect successfully CRCoV infection in dogs. We suggest that mRT-PCR would be useful and effective for monitoring CRCoV infection in various kinds of dogs.
Animals
;
Coronavirus*
;
Cough
;
Distemper Virus, Canine
;
Dogs*
;
Korea
;
Nucleocapsid Proteins
;
Orthomyxoviridae
;
Pneumonia
;
Polymerase Chain Reaction
;
Respiratory System
;
Sneezing
10.Diagnosis of Mycoplasma hyorhinis infection in pigs by PCR amplification of 16S-23S rRNA internal transcribed spacer region.
Sunhwa HONG ; Hyun A LEE ; Yungho CHUNG ; Okjin KIM
Journal of Biomedical Research 2015;16(3):104-108
Mycoplasma hyorhinis (M. hyorhinis) is considered an etiological agent of arthritis in suckling pigs. Recently, some M. hyorhinis strains were shown to produce pneumonia that is indistinguishable from the mycoplasmosis caused by M. hyopneumoniae. In this study, we developed a sensitive and specific PCR assay to detect M. hyorhinis and applied the developed PCR assay for detection of Mycoplasma infection in clinical piglets infected with M. hyorhinis. We developed a new PCR assay using a M. hyorhinis-specific primer pair, Mrhin-F and Mrhin-R, designed from the Mycoplasma 16S-23S rRNA internal transcribed spacer (ITS) region. The primers and probe for the assay were designed from regions in the Mycoplasma 16S-23S rRNA ITS unique to M. hyorhinis. The developed PCR assay was very specific and sensitive for the detection of M. hyorhinis. The assay could detect the equivalent of 1 pg of target template DNA, which indicates that the assay was very sensitive. In addition, M. hyorhinis PCR assay detected only M. hyorhinis and not any other Mycoplasma or bacterial spp. of other genera. The new developed PCR assay effectively detected M. hyorhinis infection in pigs. We suggest that this PCR assay using a M. hyorhinis-specific primer pair, Mrhin-F and Mrhin-R, could be useful and effective for monitoring M. hyorhinis infection in pigs.
Arthritis
;
Diagnosis*
;
DNA
;
Mycoplasma hyorhinis*
;
Mycoplasma Infections
;
Mycoplasma*
;
Natural Resources
;
Pneumonia
;
Polymerase Chain Reaction*
;
Swine*