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1.
The genome of Brucella melitensis   总被引:11,自引:0,他引:11  
The genome of Brucella melitensis strain 16M was sequenced and contained 3,294,931 bp distributed over two circular chromosomes. Chromosome I was composed of 2,117,144 bp and chromosome II has 1,177,787 bp. A total of 3198 ORFs were predicted. The origins of replication of the chromosomes are similar to each other and to those of other -proteobacteria. Housekeeping genes such as those that encode for DNA replication, protein synthesis, core metabolism, and cell-wall biosynthesis were found on both chromosomes. Genes encoding adhesins, invasins, and hemolysins were also identified.  相似文献   

2.
Elizabethkingia meningoseptica has been recognised as an occasional but serious opportunistic bacterial pathogen to human beings. Recently, it was frequently isolated from tiger frog, Rana tigerina rugulosa, with cataract disease, which is the most common disease of unknown aetiology of frogs in Hainan, China. The purpose of this study was to identify and characterise the bacterial strains isolated from the recent outbreaks of cataract disease in farmed tiger frog in Hainan, China, and to evaluate their pathogenicity to the frog and their sensitivity to 20 chemotherapeutic agents.The 16S rRNA gene sequences of strains W0701 (1478 bp), W0702 (1477 bp) and W0703 (1478 bp) showed 98.6–98.7% similarity with the sequence of E. meningoseptica type strain (ATCC 13253) and 99.9–100% similarity with that of E. meningoseptica NTU 870424-IL. Six strains (W0701–W0706) were selected to represent 24 isolates retrieved from six moribund frogs. The morphological, physiological and biochemical characteristics of the six representative isolates were consistent with those of E. meningoseptica strains. The organisms were only susceptible to vancomycin and moderately susceptible to cefoperazone among the 20 investigated chemotherapeutic agents. Virulence test with strain W0702 was conducted and pathogenicity (by intramuscular injection) was demonstrated in the tiger frog. In conclusion, 24 isolates obtained from frogs with cataract disease were the E. meningoseptica strains highly pathogenic to tiger frog, and this is the first report of E. meningoseptica as a pathogen for tiger frog.  相似文献   

3.
Besnoitia besnoiti was in vitro isolated during the first recorded outbreak of bovine besnoitiosis in Germany. Molecular characterization of the new isolate, named Bb-GER1, revealed almost 100% identity with other B. besnoiti isolates obtained in Portugal, Spain, Israel or South Africa, when partial sequences of the 18S ribosomal RNA gene, of the internal transcribed spacer 1 and of the 5.8S RNA gene were compared. Cystozoites obtained from skin tissue of one bull were infectious for γ-interferon knockout (GKO) mice by intraperitoneal (ip) inoculation. Tachyzoites were detected in the peritoneal cavity, spleen, liver and lung of the mice 5 days post-infection. The parasite could be maintained in GKO mice by ip inoculation for at least 5 passages. Peritoneal washings containing tachyzoites were obtained from infected mice and used to infect five cell lines (Vero, MARC-145, NA42/13, BHK21, KH-R). The best growth of tachyzoites was observed in BHK21 cells, but replication occurred to a smaller extent also in MARC-145, NA42/13 and KH-R cells. Subsequent comparative analyses revealed that after direct infection of these cell lines with cystozoites derived from bovine skin, the growth was best in NA42/13 cells. Considerable replication was also observed in the BHK21 and KH-R cell lines. Our observations on the growth characteristics of Bb-GER1 partially contrast those for other isolates. The preferential growth in particular cell lines may be characteristic for particular B. besnoiti isolates. A potential association between growth properties and differences in virulence remains to be established. This is the first in vitro isolation of B. besnoiti from cattle in Germany.  相似文献   

4.
Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma flocculare can be present in the lungs of pigs at the same time. These three mycoplasma species all require similar growth conditions and can be recovered from clinical samples using the same media. We have developed a multiplex PCR as a helpful tool for rapid differentiation of these three species in the course of isolation. Based on the 16S ribosomal DNA sequences, three different forward primers and a single reverse primer were selected. Each forward primer was compared to available mycoplasma sequences, showing the primers to be specific. The three amplification products observed of 1129 bp (M. hyorhinis), 1000 bp (M. hyopneumoniae) and 754 bp (M. flocculare) were clearly distinguishable on a 1% agarose gel. In addition, no cross-reaction with Mycoplasma hyosynoviae, another porcine mycoplasma, was noted. This multiplex PCR using the proposed set of primers is the first reported assay that allows the simultaneous identification of the different Mycoplasma species isolated from the lungs of pigs.  相似文献   

5.
A 4-year-old Icelandic horse, considered to be a mare, showed stallion-like behavior in a group of mares. On clinical examination, the horse turned out to have an enlarged erectable phallic clitoris. Ultrasound examination showed a normal-sized left ovary covered with numerous small follicular cysts and a compact testis-like tissue in place of the right ovary. The karyotype was normal for a mare (64,XX), and the horse was found to be negative for the Y chromosome–specific markers SRY, ZFY, and EIF1AY. This case indicates that the intersexual phenotype may be caused by autosomal recessive mutation, resulting in defects in cortisol biosynthesis rather than transferal of Y chromosome male–specific genes. This is the first report of an intersexual phenotype in an Icelandic horse that is likely to be a true hermaphrodite because of female sex chromosomes and a mixture of female and male gonads and external genitals.  相似文献   

6.
Rickettsia spp. are zoonotic pathogens and mainly transmitted by various arthropod vectors, such as fleas, ticks, and lice. Previous epidemiological studies indicated that ectoparasites infested on dogs or cats may be infected by Rickettsia spp., and transmit them to human beings accidentally. In this study, the prevalence of Rickettsia infection was evaluated using fleas and ticks from stray dogs and cats in Taiwan. A total of 158 pools made by 451 cat fleas (Ctenocephalides felis) from 37 dogs and 4 cats were used for analysis. Besides, 386 Rhipicephalus ticks collected from the other 62 stray dogs were included in this study. Nymphal and adult ticks were individually analyzed but larvae were separated into 21 pools for molecular detection. Partial sequencing analysis of the gltA gene was applied for Rickettsia identification. The results showed that 44.3% (70/158) of the cat flea pools were harboring Rickettsia DNA. Although 6.9% (13/187) of adult ticks were infected with Rickettsia, neither larval pools nor nymphal ticks were found to contain Rickettsia DNA. According to the results of sequencing analyses, all Rickettsia PCR-positive cat flea pools were infected with R. felis, and all Rickettsia PCR-positive adult ticks were infected with R. rhipicephali. The results of this study demonstrated that C. felis but not Rhipicephlus sanguineus (the brown dog tick) and Rh. haemaphysaloides collected from stray animals in Taiwan could be infected the zoonotic pathogen R. felis. Moreover, R. rhipicephali was only identified in adult stage of Rhipicephalus sanguineus and Rh. haemaphysaloides.  相似文献   

7.
Campylobacteriosis is a zoonosis that occurs worldwide. Infection with Campylobacter fetus (C. fetus) causes infertility and abortion in sheep and cattle. The current study focuses on the SapA gene of C. fetus that encodes surface array proteins and plays an important role in the virulence of C. fetus. The SapA-N (1398 bp) and SapA-C (1422 bp) fragments were amplified from the C. fetusSapA gene using polymerase chain reaction (PCR), and the corresponding recombinant proteins rSapA-N and rSapA-C were expressed in Escherichia. coli BL21 cells. Results of Western blotting and enzyme-linked immunosorbent assay (ELISA) showed that the immunological activity of rSapA-N was higher than that of rSapA-C (P < 0.05). Therefore, rSapA-N was selected to establish an indirect ELISA for detecting antibodies against C. fetus. The diagnostic criteria were as follows: S/P ? 0.45: positive; S/P < 0.4: negative; 0.45 > S/P ? 0.4: suspected. The specificity and sensitivity of our method were 94.3% and 88.6%, respectively. Moreover, no cross-reactions were observed between rSapA-N and serum samples that were positive for other bovine bacterial pathogens diseases such as Mycobacterium avium subspecies paratuberculosis. One hundred and two serum samples from cows that had experienced abortion were tested. Four and 2 C. fetus-positive serum samples were found among the 70 bovine brucellosis-positive samples and the 32 infectious bovine rhinotracheitis (IBR)-positive samples, respectively. The findings suggest that the rSapA-N-based ELISA method has immense potential in future applications.  相似文献   

8.
Long-term residence of the brucellae in the phagosomal compartment of host macrophages is essential to their ability to produce disease in both natural and experimental hosts. Correspondingly, the Brucella spp. appear to be well adapted to resist the multiple environmental stresses they encounter in their intracellular home. This brief review will focus on the contributions of the hfq and bacA gene products to this adaptation. Studies with Brucella hfq mutants suggest that stationary phase physiology is critical for successful long-term residence in host macrophages. Analysis of Brucella bacA mutants, on the other hand, reveal very striking parallels between the strategies employed by the rhizobia to establish and maintain protracted intracellular residence in their plant host and those used by the brucellae during their long-term survival in the phagosomal compartment of host macrophages.  相似文献   

9.
Sarcocystis tenella is a dog–sheep protozoan parasite, causing a widespread enzootic muscle parasitosis and neurological disease mainly in lambs. This parasite is pathogenic to sheep and important to the economical production of sheep. The present study was initially aimed to determine Toxoplasma gondii infection and the occurrence of co-infection with other Apicomplexa parasites in 602 Brazilian sheep. Twenty of these sheep were positive with antibodies to T. gondii by MAT and IFAT-IgG tests, positive with PCR-RFLP genotyping at multiple loci, and parasites were isolated from mice infected with sheep tissue samples. Two additional sheep born in Brazil, a 2-year-old female Polwarth (Ideal) sheep, a breed originated from Australia (#1), and a 1-year-old male Corriedale sheep, a breed originated from New Zealand and Australia (#2) were positive to T. gondii antibodies by serum tests, and PCR, but negative for bioassay in mice. In genotyping at 12 loci, sheep #1 sample and #2 presented positive results only for some markers. PCR-RFLP of 18S ribosomal RNA (18S rRNA) was performed in all 22 animals to identify the possibility of co-infection of T. gondii with other Apicomplexa parasites, such as S. tenella, Neospora caninum and Hammondia hammondi, resulting in a T. gondii profile for the first 20 animals and a unique genotyping profile for sheep #1 and #2, identical to S. tenella. The 18S rRNA PCR products (310 bp) were sequenced and blasted to GenBank database at NCBI. Both samples were identical to S. tenella 18S rRNA gene (GenBank accession number L24383-1). These results suggest the existence of co-infection of S. tenella with T. gondii in ewes from Brazil.  相似文献   

10.
猪源多杀性巴氏杆菌荚膜分型及外膜蛋白H基因序列分析   总被引:1,自引:0,他引:1  
为了解国内猪源多杀性巴氏杆菌外膜蛋白H基因的变异情况及与荚膜型之间的相关性,本试验采用PCR方法对44株猪源多杀性巴氏杆菌进行荚膜分型和ompH基因的扩增测序。结果显示,44株菌株中22株为荚膜A型,17株为荚膜B型,5株为荚膜D型;44株菌株的ompH基因开放阅读框在1 002~1 056bp之间;SignaIP 4.1预测结果显示,信号肽为N端20个氨基酸残基;ProtParam分析结果显示,成熟蛋白氨基酸残基数量在313~331aa之间,推测的分子质量在33.83~36.46ku之间。序列分析结果显示,44株菌株核苷酸同源性为86.2%~100.0%,氨基酸同源性为86.0%~100.0%;ompH基因核苷酸序列遗传进化树结果显示,荚膜A型、B型和D型菌株分别在不同的分支。试验结果表明,猪源多杀性巴氏杆菌ompH基因在不同血清型之间具有较高的同源性,与荚膜型之间存在相关性。  相似文献   

11.
为探究鲤鱼病原菌的生物学特性及耐药状况,从患病鲤鱼中分离到菌株CS126,对其进行形态学观察、生理生化性质测定、16SrDNA序列分析及药敏性测定。结果显示,菌株CS126为革兰氏阴性菌,不产生吲哚、不具有动力性,分解半乳糖、甘露醇,VP试验、赖氨酸脱羧酶、氧化酶阴性,七叶苷阳性。16SrDNA序列长度为1 459bp,GenBank登录号为KJ942580,与GenBank中杀鲑气单胞菌的相似性高达100%,进化树显示与杀鲑气单胞菌杀鲑亚种聚为一分支。这表明菌株CS126为杀鲑气单胞菌杀鲑亚种。该菌株对菌必治、诺氟沙星、氟苯尼考等8种药物高度敏感,但对氨苄西林、氨苄西林/舒巴坦、甲氧卞胺嘧啶和磺胺异恶唑等耐药。本试验结果为鱼类杀鲑气单胞菌的快速鉴定及鱼类疾病诊治提供参考依据。  相似文献   

12.
The monogenean Gyrodactylus salaris Malmberg, 1957 is considered one of the most important parasites of wild salmonids in the European Community due to the heavy ecological and economical damage it has inflicted on Atlantic salmon (Salmo salar) parr populations. Rainbow trout (Oncorhynchus mykiss) is susceptible to G. salaris and can act as a suitable carrier host and, consequently, its trade in EU territory is restricted in relation to the status of “recognized free” zones. Despite the economic importance of rainbow trout farming in Italy, information on the Italian gyrodactylid fauna is lacking and prior to this study, G. salaris had not been officially reported. During a routine health examination of farmed rainbow trout stock throughout Central and Northern Italy in 2004–2005, five fish farms were found to be infected with G. salaris alongside three other gyrodactylids. Morphological and molecular characterisation confirmed the presence of G. salaris, Gyrodactylus teuchis Lautraite, Blanc, Thiery, Daniel et Vigneulle, 1999 and Gyrodactylus derjavinoides Malmberg, Collins, Cunningham et Jalali, 2007, while Gyrodactylus truttae Gläser, 1974 was identified by morphological analysis only. The findings from this study extend the distribution of G. salaris within Europe and highlight the importance of the rainbow trout trade in its dissemination.  相似文献   

13.
The prevalence of Bartonella spp. in wild rodents was studied in 19 geographical locations in Israel. One hundred and twelve rodents belonging to five species (Mus musculus, Rattus rattus, Microtus socialis, Acomys cahirinus and Apodemus sylvaticus) were included in the survey. In addition, 156 ectoparasites were collected from the rodents. Spleen sample from each rodent and the ectoparasites were examined for the presence of Bartonella DNA using high resolution melt (HRM) real-time PCR. The method was designed for the simultaneous detection and differentiation of eight Bartonella spp. according to the nucleotide variation in each of two gene fragments (rpoB and gltA) and the 16S–23S intergenic spacer (ITS) locus, using the same PCR protocol which allowed the simultaneous amplification of the three different loci. Bartonella DNA was detected in spleen samples of 19 out of 79 (24%) black rats (R. rattus) and in 1 of 4 (25%) Cairo spiny mice (A. cahirinus). In addition, 15 of 34 (44%) flea pools harbored Bartonella DNA. Only rat flea (Xenopsyla cheopis) pools collected from black rats (R. rattus) were positive for Bartonella DNA. The Bartonella sp. detected in spleen samples from black rats (R. rattus) was closely related to both B. tribocorum and B. elizabethae. The species detected in the Cairo spiny mouse (A. cahirinus) spleen sample was closely related to the zoonotic pathogen, B. elizabethae. These results indicate that Bartonella species are highly prevalent in suburban rodent populations and their ectoparasites in Israel. Further investigation of the prevalence and zoonotic potential of the Bartonella species detected in the black rats and the Cairo spiny mouse is warranted.  相似文献   

14.
To ascertain the infection rate for tick-borne pathogens in Zambia, an epidemiological survey of Theileria parva, Babesia bigemina and Anaplasma marginale in traditionally managed Sanga cattle was conducted using PCR. Of the 71 native Zambian cattle, 28 (39.4%) were positive for T. parva, 16 (22.5%) for B. bigemina and 34 (47.9%) for A. marginale. The mixed infection rate in cattle was 8.5% (6/71), 16.9% (12/71), 7.0% (5/71) and 2.8% (2/71) for T. parva/B. bigemina, T. parva/A. marginale, B. bigemina/A. marginale and T. parva/B. bigemina/A. marginale, respectively.To predict the risk for transmission of tick-borne pathogens from ticks to cattle, a total of 74 Rhipicephalus appendiculatus ticks were collected from a location where cattle had been found positive for T. parva. Of the ticks collected, 10 (13.5%) were found to be PCR-positive for T. parva. The results suggest that the infection rate for tick-borne pathogens was relatively high in Sanga cattle and that adult R. appendiculatus ticks were highly infected with T. parva.  相似文献   

15.
Vibrio tasmaniensis, Vibrio splendidus and Vibrio neptunius species were distributed worldwide and associated with aquaculture and have been reported as the cause of diseases in aquatic organisms. Polyphasic analyses for bacterial identification are not feasible for routine diagnostic because of the time involved. The aim of this study is to design three PCR primer sets that can assist with fast detection of these species. They were designed from the 16S ribosomal RNA gene, and PCR conditions were found. Each PCR test successfully identified all the tested strains of each target species. The combined specificity of V. tasmaniensis and V. splendidus primer sets offered the best coverage (86%) in terms of separating target organisms from other related species. The primer set of V. tasmaniensis showed a lower sensitivity limit (500 fg of DNA) than the V. splendidus set (1 pg) and both sets gave positive amplification using homogenized tissues from inoculated clams, with 102 and 104 cfu/g of clam, respectively. The primer set of V. neptunius was highly specific, showing only cross-reaction with V. parahaemolyticus species from 44 tested species. Its sensitivity limit was 100 pg of DNA. A small number of biochemical tests were proposed concurrently with the PCR to differentiate the cross-reacting bacteria. The time of detection of the three tested species was reduced and the further affected animals can be diagnosed in a rapid fraction of time. The detection of virulent strains of V. tasmaniensis pointed to the risk of mollusc culture outbreaks.  相似文献   

16.
为预测弓形虫细胞核因子3(TgNF3)基因作为抗弓形虫疫苗候选抗原的可能性,本试验以弓形虫RH株的总RNA为模板,应用逆转录-聚合酶链式反应(RT-PCR)扩增TgNF3基因并连接至pMD18-T载体。筛选阳性克隆后,测序鉴定。将测序结果正确的核苷酸序列翻译成氨基酸序列,运用生物信息学软件分析TgNF3蛋白的结构,预测抗原表位。结果显示,TgNF3基因长约950bp。TgNF3蛋白中包含1个跨膜结构域,9个α-螺旋区,4个β-折叠区,8个亲水区域和10个柔性区域,并预测含有9个线性B细胞抗原表位。结果表明,TgNF3蛋白可能具有良好的免疫原性,为研制以TgNF3为抗原的弓形虫亚单位疫苗及表位疫苗奠定了基础。  相似文献   

17.
水貂奇异变形杆菌的分离鉴定及16S rRNA基因序列分析   总被引:1,自引:1,他引:0  
从辽宁某貂场发病水貂中分离到1株致病菌,命名为PMSD株,通过形态学观察和生化试验等常规鉴定发现符合奇异变形杆菌特性,进一步经VITEK 2Compact 30全自动细菌鉴定及药敏分析系统鉴定该株细菌为奇异变形杆菌。药敏试验结果显示PMSD株对氨基糖苷类药物、喹诺酮类药物等敏感,而对β-内酰胺类药物和磺胺类药物等不敏感。以细菌16SrRNA基因为模板应用通用引物进行PCR扩增,得到PMSD株的16SrRNA基因序列,长约1 504bp,提交到GenBank中,登录号:KM229530。将该序列与GenBank中序列进行BLAST比对,结果发现与其匹配度最高的均是奇异变形杆菌各株系的16SrRNA序列,均高达99%以上。选取其中前20株作为参考序列,运用生物学软件构建系统发育树并进行同源性比对,结果表明,分离菌PMSD株与20个代表菌株的同源性为98.9%~99.7%,其中与BB2000株、HI4320株、B1株和FCC141株同源性最高,为99.7%。本研究为预防和控制奇异变形杆菌引起的水貂疾病奠定了一定的基础。  相似文献   

18.
为了解动物沙门氏菌的流行情况和药物敏感性及氟苯尼考耐药株的耐药基因分布,本试验对临床上疑似患沙门氏菌病的病料进行病原分离和细菌的多重PCR鉴定;采用K-B法测定分离株对23种抗菌药物的敏感性;选择氟苯尼考耐药菌株扩增floR、fexA、fexB、cfr和pexA基因。结果显示,共鉴定出61株沙门氏菌,其中肠炎沙门氏菌10株,鸡白痢沙门氏菌12株,鼠伤寒沙门氏菌39株。所有菌株对青霉素、红霉素、万古霉素耐药,90.16%对6种及6种以上抗菌药耐药。floR基因广泛存在于鼠伤寒沙门氏菌氟苯尼考耐药菌株中(8/12,66.67%),未发现其他耐药基因。研究结果表明鼠伤寒沙门氏菌是鹅源分离株中的优势血清型;floR基因主要介导沙门氏菌对氟苯尼考耐药性,但可能还存在其他机制。  相似文献   

19.
Bartonellae are gram-negative facultative intracellular alpha-proteobacteria from the family Bartonellaceae. The natural history of bartonellae consists of a reservoir/host, which is a vertebrate with chronic intravascular infection with sustained bacteremia, and a vector (usually an arthropod) that transfers the bacteria from the reservoir to a susceptible yet uninfected host. In order to reveal the sources and reservoirs of Bartonella infection in animals and vectors in New Caledonia, we collected the blood samples of 64 dogs, 8 cats, 30 bovines, 25 horses and 29 wild deer Cervus timorensis russa and 308 associated blood-sucking parasites (14 keds Hippobosca equina, 258 ticks (22 Rhipicephalus microplus, 235 Rhipicephalus sanguineus, and 1 Haemaphysalis longicornis), 12 fleas Ctenocephalides felis and 24 dog lice Trichodectes canis). We isolated ten strains of Bartonella: four Bartonella henselae from cats and six Bartonella chomelii from cattle. The strains were characterized by sequencing of five genes (16S, ITS, rpoB, gltA and ftsZ). The six strains isolated from cattle were close to the reference strain of B. chomelii and were, probably, imported from France with cattle of Limousin race. PCR showed that 35% of keds collected from deer and 31% of deer were infected by B. aff. schoenbuchensis; all other samples were negative. Our data confirmed that in New Caledonia, as in other regions of the world, cats are the major reservoirs of B. henselae. We also confirmed that Hippoboscidae flies may serve as the vectors of ruminant-associated bartonellae.  相似文献   

20.
Antimicrobial therapy is a useful tool to control bovine mastitis caused by Staphylococcus aureus, as consequence an increase in staphylococci resistant cases has been registered. Alternative strategies are desirable and bacteriocins represent attractive control agents to prevent bovine mastitis. The aim of this work was to evaluate the activity of five bacteriocins synthesized by Bacillus thuringiensis against S. aureus isolates associated to bovine mastitis. Fifty S. aureus isolates were recovered from milk composite samples of 26 Holstein lactating cows from one herd during September 2007 to February 2008 in México and susceptibility of those isolates to 12 antibiotics and 5 bacteriocins from B. thuringiensis was evaluated. S. aureus isolates were mainly resistant to penicillin (92%), dicloxacillin (86%), ampicillin (74%) and erythromycin (74%); whereas susceptibility to gentamicin, trimethoprim and tetracycline was detected at, respectively, 92%, 88%, and 72%. All S. aureus isolates showed susceptibility to the five bacteriocins synthesized by B. thuringiensis, mainly to morricin 269 and kurstacin 287 followed by kenyacin 404, entomocin 420 and tolworthcin 524. Our results showed that S. aureus isolates had differences in the antimicrobial resistance patterns and were susceptible to bacteriocins produced by B. thuringiensis, which could be useful as an alternative method to control bovine mastitis.  相似文献   

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