Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis   总被引：1，自引：0，他引：1  

Alhassan A  Thekisoe OM  Yokoyama N  Inoue N  Motloang MY  Mbati PA  Yin H  Katayama Y  Anzai T  Sugimoto C  Igarashi I 《Veterinary parasitology》2007,143(2):155-160

Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid method whereby DNA is amplified with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specifically designed primers and a DNA polymerase with strand displacement activity. In this study, we used LAMP primer sets designed from EMA-1 and Bc 48 genes for detection of Theileria equi and Babesia caballi infections, respectively. These primer sets specifically amplified DNA of the respective parasites. Both primer sets amplified T. equi and B. caballi up to 10(-6) dilution of 10-fold serially diluted samples. Furthermore, DNA extracted from blood collected from a horse experimentally infected with T. equi was amplified by a T. equi LAMP primer set from days 2 to 35 post-infection, demonstrating the high sensitivity of these primers. Of 55 samples collected from China, 81.8% and 56.3% were positively detected by LAMP for T. equi and B. caballi infections, respectively. In contrast, 91.8% and 45.9% of the 37 samples collected from South Africa were LAMP positive for T. equi and B. caballi, respectively. These results suggest that LAMP could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis.  相似文献   

Genetic diversity of benign Theileria parasites of cattle in the Okinawa Prefecture     

Zakimi S  Kim JY  Oshiro M  Hayashida K  Fujisaki K  Sugimoto C 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(12):1335-1338

Benign Theileria parasites of cattle distributed in the Okinawa prefecture were characterized by allele-specific polymerase chain reaction (PCR) and DNA sequence analysis of the major piroplasm surface protein (MPSP) gene. Using universal or allele-specific primer sets, parasite DNA was amplified in 31 out of 48 blood samples obtained from beef cattle. Among the positive cases, mixed infections involving various combinations of I-, C-, and B-type parasites were detected in 24 (77.4%) samples. Phylogenetic analysis based on the MPSP gene sequences revealed that parasites with the MPSP types 1-5 and 7, exist within the Okinawa prefecture.  相似文献   

Effects of hyaluronic acid on Ca(2+) influx, lactate dehydrogenase activity, and cyclic AMP synthesis in canine ejaculated sperm during In Vitro capacitation     

Kawakami E  Nishizono Y  Miyata C  Hirano T  Hori T  Tsutsui T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(2):119-123

The effects of hyaluronic acid, which comprises the cumulus intercellular matrix, on Ca(2+) influx, lactate dehydrogenase (LDH) activity, and cyclic AMP synthesis in canine sperm during capacitation was investigated. Ejaculated sperm were collected from 10 Beagle dogs and the sperm were incubated for 4 hr in Eagle's MEM containing 10 microg/ml of hyaluronic acid. The percentages of actively motile sperm, hyperactivated sperm (HA-sperm), acrosome-reacted sperm (AR-sperm), and sperm labeled with fluoresceinated Ca(2+) indicator (Ca(2+)-labeled sperm) were evaluated to assess Ca(2+) influx into the sperm. LDH activity and cAMP concentration were measured in homogenized sperm. The mean percentages of motile sperm, HA-sperm, and Ca(2+)-labeled sperm in the MEM containing hyaluronic acid were higher than in the control medium (P<0.05, 0.05, and 0.01, respectively), but there was no difference between the percentages of AR-sperm. Mean LDH activity and mean cAMP concentration were also significantly higher than the control values (P<0.05). The percentages of HA-sperm correlated with those of Ca(2+)-labeled sperm (r(2)=0.810). The results indicate that hyaluronic acid increases Ca(2+) influx, LDH activity, and cAMP synthesis in canine ejaculated sperm during capacitation in vitro.  相似文献   

Innate immune response of mammary gland induced by intramammary infusion of Bifidobacterium breve in lactating dairy cows     

Hajime NAGAHATA  Ayumi MORIYAMA  Chika SAWADA  Yukiko ASAI  Chihiro KOKUBU  Satoshi GONDAIRA  Hidetoshi HIGUCHI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2020,82(12):1742

This study aimed to evaluate innate immune responses of mammary glands induced by intramammary infusion of Bifidobacterium breve in dairy cows. Somatic cell counts in quarters of cows showed a marked increase following B. breve infusion on days 1 and 2. Opsonized-stimulated chemiluminescence response in quarter milk was significantly (P<0.05) increased by B. breve infusion on days 1 to 3 compared to that of pre-infusion. Lactoferrin concentrations in B. breve-infused quarter milk increased significantly (P<0.05) on days 2 to 4 and 6 compared to those of pre-infusion. IgG and IgA concentrations in B. breve-infused quarters significantly (P<0.05) increased on days 2 to 4 for IgG and days 3, 4, 6 and 8 for IgA compared to those of pre-infusion. Interleukin (IL)-1β and IL-8 mRNA levels in somatic cells from B. breve-infused quarters were significantly (P<0.05) upregulated on day 1 compared to those on days 0 and 14. Conversely, IL-6 mRNA levels in somatic cells from B. breve-infused quarters on days 0, 1 and 14 and NF-κB mRNA levels on day 0 were significantly (P<0.05) down-regulated compared to those of control. IL-1β, tumor necrosis factor (TNF)-α and IL-6 concentrations increased on days 1, 3 and 7 after B. breve infusion in quarters. Intramammary infusion of B. breve (3 × 10⁹ cfu) induces a massive influx of leukocytes and enhances innate immune response in mammary glands. This event may contribute to the enhancing host defense in the mammary gland.  相似文献   

Molecular detection of zoonotic tick-borne pathogens from ticks collected from ruminants in four South African provinces     

Khethiwe MTSHALI  Zamantungwa T. H. KHUMALO  Ryo NAKAO  Dennis J. GRAB  Chihiro SUGIMOTO  Oriel M. M. THEKISOE 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(12):1573-1579

Ticks carry and transmit a remarkable array of pathogens including bacteria, protozoa and viruses, which may be of veterinary and/or of medical significance. With little to no information regarding the presence of tick-borne zoonotic pathogens or their known vectors in southern Africa, the aim of our study was to screen for Anaplasma phagocytophilum, Borrelia burgdorferi, Coxiella burnetii, Rickettsia species and Ehrlichia ruminantium in ticks collected and identified from ruminants in the Eastern Cape, Free State, KwaZulu-Natal and Mpumalanga Provinces of South Africa. The most abundant tick species identified in this study were Rhipicephalus evertsi evertsi (40%), Rhipicephalus species (35%), Amblyomma hebraeum (10%) and Rhipicephalus decoloratus (14%). A total of 1634 ticks were collected. DNA was extracted, and samples were subjected to PCR amplification and sequencing. The overall infection rates of ticks with the target pathogens in the four Provinces were as follows: A. phagocytophilum, 7%; C. burnetii, 7%; E. ruminantium, 28%; and Rickettsia spp., 27%. The presence of B. burgdorferi could not be confirmed. The findings of this study show that zoonotic pathogens are present in ticks in the studied South African provinces. This information will aid in the epidemiology of tick-borne zoonotic diseases in the country as well as in raising awareness about such diseases in the veterinary, medical and tourism sectors, as they may be the most affected.  相似文献   

Pathogenicity of border disease virus FNK2012-1 strain isolated from a pig in the natural host,sheep     

Tomokazu TAMURA  Junki MINE  Shiho TORII  Yuri FUJIMOTO  Masatoshi OKAMATSU  Yoshihiro SAKODA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(3):341-343

A first isolation of border disease virus (BDV) in Japan was from a pig on a farm without keeping any ruminants. Our previous study showed that this BDV, termed the FNK2012-1 strain, replicated inefficiently in swine-derived cells compared with those of ruminant origin. Pigs inoculated with this virus showed neither clinical symptoms nor viremia. In this study, we evaluated the pathogenicity of the FNK2012-1 strain in sheep, its natural host. The inoculated sheep showed clinical symptoms and transient viremia. Seroconversion was observed in the inoculated sheep. These results suggest that the FNK2012-1 strain was introduced from sheep and has not yet adapted to swine. Therefore, surveillance of border disease in Japan is necessary among both the swine and ruminant populations.  相似文献   

Developmental capacity and implantation potential of the embryos with multinucleated blastomeres     

Akiyoshi EGASHIRA  Nobuhiko YAMAUCHI  Keiko TANAKA  Chihiro MINE  Hitomi OTSUBO  Masao MURAKAMI  Md. Rashedul ISLAM  Misako OHTSUKA  Naomi YOSHIOKA  Takashi KURAMOTO 《The Journal of reproduction and development》2015,61(6):595-600

The presence of multinucleated blastomeres (MNBs) in embryos is associated with poor developmental competence in assisted reproductive technologies. This phenomenon is observed not only in humans but also in other animal species. The purpose of the present study was to investigate the characteristics of embryos with MNBs (MNB embryos) that could be utilized in embryo transfer. The developmental rate of MNB embryos to the blastocyst stage (50.8%) was significantly lower than that of normal embryos (73.3%) (P < 0.05). The clinical pregnancy rates of fresh embryo transfer (ET) using day 2 or day 3 embryos were significantly lower in MNB embryos (5.1%) compared with normal embryos (24.0%) (P < 0.05). In the case of frozen-thawed ET using a single vitrified/warmed blastocyst, however, the clinical pregnancy rate of MNB embryos was close to that of normal embryos (59.1% vs. 52.8%). Thus, the findings of the present study suggest that the frozen-thawed ET of MNB embryos might improve the potential for implantation followed by successful pregnancy.  相似文献   

Unrecognized difficult airway management during anesthesia in two brachycephalic dogs with narrow cricoid cartilage     

Jun TAMURA  Norihiko OYAMA  So MATSUMOTO  Ryo OWAKI  Kenji HOSOYA  Masahiro OKUMURA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(2):234

Difficulty in airway management during anesthesia was noted in a 10-year-old, castrated, male Pekingese dog and a 13-year-old male French Bulldog. They showed strong resistance during tracheal tube insertion through the subglottic lumen. Therefore, the airway was secured by using a small endotracheal tube or supraglottic airway device. Computed tomography scan revealed a markedly narrower vertical dimension of the cricoid cartilage compared to that seen in common brachycephalic breeds. Posterior glottis was relatively more accessible for translaryngeal intubation in the present cases. Our findings showed that brachycephalic airway syndrome may be associated with narrow cricoid cartilage. To the best of our knowledge, this is the first clinical case report of airway management during anesthesia in dogs with narrow cricoid cartilage.  相似文献   

Production of alpha 1,3-galactosyltransferase-deficient pigs     

Phelps CJ  Koike C  Vaught TD  Boone J  Wells KD  Chen SH  Ball S  Specht SM  Polejaeva IA  Monahan JA  Jobst PM  Sharma SB  Lamborn AE  Garst AS  Moore M  Demetris AJ  Rudert WA  Bottino R  Bertera S  Trucco M  Starzl TE  Dai Y  Ayares DL 《Science (New York, N.Y.)》2003,299(5605):411-414

The enzyme alpha1,3-galactosyltransferase (alpha1,3GT or GGTA1) synthesizes alpha1,3-galactose (alpha1,3Gal) epitopes (Galalpha1,3Galbeta1,4GlcNAc-R), which are the major xenoantigens causing hyperacute rejection in pig-to-human xenotransplantation. Complete removal of alpha1,3Gal from pig organs is the critical step toward the success of xenotransplantation. We reported earlier the targeted disruption of one allele of the alpha1,3GT gene in cloned pigs. A selection procedure based on a bacterial toxin was used to select for cells in which the second allele of the gene was knocked out. Sequencing analysis demonstrated that knockout of the second allele of the alpha1,3GT gene was caused by a T-to-G single point mutation at the second base of exon 9, which resulted in inactivation of the alpha1,3GT protein. Four healthy alpha1,3GT double-knockout female piglets were produced by three consecutive rounds of cloning. The piglets carrying a point mutation in the alpha1,3GT gene hold significant value, as they would allow production of alpha1,3Gal-deficient pigs free of antibiotic-resistance genes and thus have the potential to make a safer product for human use.  相似文献   

Insect antifeedants from tropical plants: structures of dumnin and dumsenin     

Nihei K  Asaka Y  Mine Y  Ito C  Furukawa H  Ju-Ichi M  Kubo I 《Journal of agricultural and food chemistry》2004,52(11):3325-3328

Two novel A-seco limonoids, dumnin and dumsenin, were isolated from the methanolic extract of Croton jatrophoides by bioassay-guided fractionation, and the structures were determined by nuclear magnetic resonance, circular dichroism, and mass spectrometry experiments. These compounds showed potent antifeedant activity (PC(50) 相似文献