Serological diagnosis of enzootic pneumonia of swine by a double-sandwich enzyme-linked immunosorbent assay using a monoclonal antibody and recombinant antigen (P46) of Mycoplasma hyopneumoniae     

Okada M  Asai T  Futo S  Mori Y  Mukai T  Yazawa S  Uto T  Shibata I  Sato S 《Veterinary microbiology》2005,105(3-4):251-259

To facilitate the control of enzootic pneumonia (EP) of swine caused by Mycoplasma hyopneumoniae, the complement fixation (CF) test has been used for the detection of M. hyopneumoniae antibodies. However, the CF test is a cumbersome and time-consuming technique and cross-reactivity are major drawbacks associated with this method. To circumvent these drawbacks, we have developed a double-sandwich enzyme-linked immunosorbent assay (ELISA), consisting of purified monoclonal antibody (Mab) against the 46 kDa surface antigen (P46) of M. hyopneumoniae and recombinant P46 protein expressed in Escherichia coli, for the detection of antibodies to M. hyopneumoniae in serum samples from pigs experimentally inoculated with M. hyopneumoniae and from naturally infected pigs, and compared the practical usefulness of ELISA using the CF test. In experimentally inoculated pigs, the CF and ELISA antibodies were detected at almost the same time, and a good correlation was demonstrated between the CF test and the ELISA. In a survey conducted on field samples, the seropositivity by ELISA in pigs of age 2-6 months was increased. At the time of slaughter, approximately 80% of the animals were seropositive for ELISA. However, a gradual decrease in the prevalence of ELISA positive samples was observed in sows with increasing parity. No correlation was seen between the results obtained with the two methods in the clinical samples. The CF test appears to have limited value for the diagnosis of EP in conventional herds because nonspecific reactions were frequently observed. Therefore, this ELISA is a useful alternative to the CF test currently used for the diagnosis of EP.  相似文献   

<Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated transformation as a tool for random mutagenesis of <Emphasis Type="Italic">Colletotrichum trifolii</Emphasis>     

Hiroyuki?Takahara  Gento?Tsuji  Yasuyuki?Kubo  Mikihiro?Yamamoto  Kazuhiro?Toyoda  Yoshishige?Inagaki  Yuki?Ichinose  Tomonori?ShiraishiEmail author 《Journal of General Plant Pathology》2004,70(2):93-96

We transformed Colletotrichum trifolii, the causal agent of alfalfa anthracnose, using Agrobacterium tumefaciens as a new tool for random insertional mutagenesis. Fungal spores of C. trifolii were transformed with T-DNA including the hygromycin phosphotransferase gene (hph). Southern analysis showed that every randomly selected transformant had a unique hybridization pattern of T-DNA, suggesting that the T-DNA was randomly integrated into the fungal genome. More significantly, about 75% of transformants had a single copy of the T-DNA. The results demonstrate that insertional mutagenesis via A. tumefaciens is a useful tool for studying the function of C. trifolii genes.  相似文献   

Isolation and expression of five-amino-acid-deleted variant of feline hepatocyte growth factor (HGF) cDNA     

Miyake M  Yaguchi T  Saze K  Wang J  Ogawa T  Endo Y  Suzuta Y  Okazaki M  Haga Y  Waki T  Takahashi SY  Yamamoto Y  Iwabuchi S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(1):9-14

Hepatocyte growth factor (HGF) is a pleiotropic cytokine that stimulates a wide array of cellular targets, including hepatocytes and other epithelial cells, melanocytes, endothelial and hematopoietic cells. We have cloned a different form of cDNA, with a deletion of 15 base pairs predicted to result in the loss of 5 amino acids from the first kringle domain. To investigate the biological activity, original and deleted variant of feline HGF cDNAs were transiently expressed in COS-7 cells. Both recombinant feline HGFs showed almost the same dose-response curves in the stimulation of the growth of BNL CL.2 cells (a mouse hepatocyte cell line) and scatter activity of Madin-Darby canine kidney (MDCK) cells. The findings reported here suggest that the deleted variant of feline HGF has almost the same biological activity as the original in terms of the proliferation and scatter activity.  相似文献   

Allergen-specific immunotherapy induces Th1 shift in dogs with atopic dermatitis     

Shida M  Kadoya M  Park SJ  Nishifuji K  Momoi Y  Iwasaki T 《Veterinary immunology and immunopathology》2004,102(1-2):19-31

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Effects of beta-thujaplicin on anti-Malassezia pachydermatis remedy for canine otitis externa     

Nakano Y  Wada M  Tani H  Sasai K  Baba E 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(12):1243-1247

The antifungal activity of beta-thujaplicin was evaluated against 51 Malassezia pachydermatis strains isolated from canine ear canals with or without otitis externa. For comparison, sensitivity tests were performed on M. pachydermatis isolates for nystatin, ketoconazole, and terbinafine HCl, all clinically available antifungal agents. The minimal inhibition concentrations over 50% of the tested isolates (MIC50) were 3.13 microg/ml for beta-thujaplicin and nystatin, 0.016 microg/ml for ketoconazole, and 1.56 microg/ml for terbinafine HCl. The antifungal effect for M. pachydermatis of beta-thujaplicin compared favorably with commercial antifungal agents. None of the 51 M. pachydermatis isolates showed resistance against any of the tested antibiotics investigated in this study. Ten representative isolates of M. pachydermatis were subcultured for 30 generations at concentrations close to the MIC levels of beta-thujaplicin, nystatin, ketoconazole, and terbinafine HCl, and examined to determine whether they had acquired resistance to each drug. As a result, M. pachydermatis was found to achieve resistance more easily for ketoconazole and terbinafine HCl than for beta-thujaplicin or nystatin. The MIC50 of beta-thujaplicin did not change during the course of subculture, and it is thought that the potential development of a resistant strain is low, even with continuous infusion for otitis externa therapy. beta-Thujaplicin is an inexpensive and safe treatment with anti-inflammatory and deodorant effects that can be recommended as an effective remedy for canine otitis externa.  相似文献   

Nonsuppurative myocarditis caused by porcine circovirus type 2 in a weak-born piglet     

Mikami O  Nakajima H  Kawashima K  Yoshii M  Nakajima Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(7):735-738

Reproductive failure associated with porcine circovirus type 2 (PCV2) was observed in a farm, and a weak-born 8-day-old piglet was examined pathologically. Focal to locally extensive lesions, including infiltration of macrophages and lymphocytes, fibrosis and degeneration of the myocardium were observed in the heart. PCV2 antigens and nucleic acids were detected in degenerated cardiomyocytes and macrophages infiltrating the heart by immunohistochemical staining and in situ hybridization. Depletion of lymphocytes with occasional infiltration of multinucleated giant cells was seen in the lymphoid organs and PCV2 antigens were demonstrated in histiocytic cells. Crystalline arrays of viral particles were observed in infiltrated macrophages in the heart and, rarely, in cardiomyocytes by electron microscopy. Although myocarditis is a common finding in aborted or stillborn piglets in reproductive failure due to PCV2, it was also observed in the 8-day-old piglet with PCV2 association.  相似文献   

Molecular cloning of canine nm23 cDNAs and their expression in normal and tumor tissues     

Takahashi M  Une R  Fukushima K  Fujiki M  Misumi K  Miyoshi N  Endo Y  Ohishi A  Akuzawa M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(8):837-841

Two canine nm-23 cDNAs, designated as nm23-C1 and -C2, were isolated and characterized. Both have a putative open reading frame consisting of 459-bp encoding 152 amino acids and are highly similar to human, mouse and rat homologues. To understand the potential role of nm23-C1 and -C2 in the development of mammary gland tumors (MGT), we analyzed the mRNA expression in 14 MGT samples by RT-PCR. The samples were divided into categories according to their histopathology (benign/malignant) and metastasis. No significant difference in the mRNA expression levels of either nm23-C1 or -C2 were observed between the benign and malignant groups or the metastatic and non-metastatic groups. These results suggest that nm23-C1 and -C2 are not related to the establishment of malignancy and metastatic lesions in canine MGT cases.  相似文献   

Canine mdr1 gene mutation in Japan     

Kawabata A  Momoi Y  Inoue-Murayama M  Iwasaki T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(11):1103-1107

Frequency of the 4-bp deletion mutant in canine mdr1 gene was examined in 193 dogs of eight breeds in Japan. The mutant allele was found in Collies, Australian Shepherds, and Shetland Sheepdogs, where its respective frequencies were 58.3%, 33.3%, and 1.2%. The MDR1 protein was detected on peripheral blood mononuclear cells (PBMC) from a MDR1/MDR1 dog, but not on PBMC from a mdr1-1Delta/mdr1-1Delta Collie. Rhodamine 123 was extruded from MDR1/MDR1 lymphocytes. That excretion was inhibited by a MDR1 inhibitor, verapamil. On the other hand, Rh123 excretion was not observed from lymphocytes derived from a mdr1-1Delta/mdr1-1Delta Collie. These results indicated that the mutant mdr1 allele also existed in Collie-breed dogs in Japan at high rates and that mdr1-1Delta /mdr1-1Delta dogs have no functional MDR1.  相似文献   

Effects of supplementation with cellooligosaccharides on growth performance of weaned calves on pasture          下载免费PDF全文

Kyoko Kido  Shigeki Tejima  Haruka Nagayama  Yutaka Uyeno  Yasuyuki Ide  Shiro Kushibiki 《Animal Science Journal》2016,87(5):661-665

We conducted two feeding experiments to evaluate the effects of cellooligosaccharide (CE) supplementation on growth performance in grazing beef calves. Calf sex and age and duration of the experimental period differed between the experiments. Experiment 1 (10 weeks) used 5.6‐month‐old castrated males; Experiment 2 (13 weeks) used 3.9‐month‐old females. Eight Japanese Black calves were assigned to either a control group (CON) or an experimental group (CE) fed CE at a rate of 10 g/day mixed with concentrate. Calves were stocked in fields in which Kentucky bluegrass was dominant. In both experiments, average daily gain tended to be greater in CE than in CON, especially in late stages of experiments, but no significant differences in body weight changes were observed. In Experiment 2, final heart girth was larger in CE than in CON calves. We monitored rumen microbial community composition in Experiment 2 and found increases in fibrolytic bacteria and methanogenic archaea in CE calves, but the overall microbial composition did not differ between the groups. Our results suggest that supplementation with CE may positively affect growth performance in weaned calves on pasture, but longer‐term CE supplementation may be required to exhibit the effect.  相似文献   

Dynamics of infection-related morphogenesis and pathogenesis in Colletotrichum orbiculare     

Yasuyuki Kubo  Yoshitaka Takano 《Journal of General Plant Pathology》2013,79(4):233-242

Colletotrichum orbiculare (syn. C. lagenarium) is the causal agent of anthracnose disease on cucurbit plants. This fungus forms dome-shaped, melanized appressoria as a host invasion structure. Strain 104-T (MAFF240422) of C. orbiculare, which was originally isolated from a cucumber plant in 1951 by Dr. Yasumori, Kyoto University, has proven to be an excellent experimental model for the study of fungal pathogenesis and morphogenesis because of its stable pathogenicity and synchronous infection-related morphogenesis. This review considers the discoveries made during 60 years of study on C. orbiculare. In particular, we focus on advances made in the last two decades, which have provided a basis for the molecular analysis not only of fungal morphogenesis, but also of plant–microbe interactions, including plant immunity to adapted and nonadapted Colletotrichum fungi. This substantial body of innovative research was originated by the Phytopathological Society of Japan and represents a major contribution to the international research communities working on plant pathology, plant–microbe interactions, and fungal molecular genetics. This review deals with the past achievements and future prospects in the study of Colletotrichum biology, focusing on the molecular genetics of C. orbiculare with regard to four aspects: (1) metabolic and functional development of infection structures, (2) signaling pathways required for fungal pathogenesis, (3) pathogen-associated molecular patterns (PAMPs) and host basal resistance, and (4) establishment of host specificity.  相似文献