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1.
The effect of cultural conditions on the production of leukotoxin by biotypes A and B of F. necrophorum was investigated. Biotypes A and B were grown in prereduced, anaerobically sterilized, brain-heart infusion (BHI) broth. The average leukotoxin titer of culture supernatant was 18 times higher from biotype A strains than from biotype B strains. Leukotoxin activity peaked during the late-log and early-stationary phases of growth, then declined precipitously in both biotypes. F. necrophorum biotype A was grown in different media (BHI, liver infusion, and Eugon broths), at various pH (6.6, 7.3, 7.7, and 8.2), incubation temperatures (30, 35, 39, and 43 degrees C), redox potentials (-352 to +375 mV), and iron concentrations (less than 0.2, 4.2, 42.1, and 361.4 microM). Anaerobic BHI broth with pH from 6.6 to 7.7 at 39 degrees C incubation temperature supported maximal F. necrophorum growth and leukotoxin production. The optimum redox potential for F. necrophorum growth was in the range of -230 to -280 mV. However, the presence of titanium III citrate or dithiothreitol (7.78 mM) in the medium decreased (P less than 0.05) the leukotoxicity of F. necrophorum. Low iron concentration (less than 0.2 microM) decreased (P less than 0.05) growth rate but not leukotoxin activity of F. necrophorum, whereas high iron concentration inhibited the leukotoxin activity. 相似文献
2.
The growth rates of 16 isolates of Tritrichomonas foetus from three distinct geographic regions were investigated in modified Diamond's medium, liver infusion broth medium and a commercially available culture kit. While some differences in growth characteristics were detected for different isolates and in the three different media, all isolates grew. Trichomonads reached peak concentrations from an initial concentration of 10(4) trichomonads/ml on Days 2, 3 and 4 in modified Diamond's medium, on Days 2-6 (excluding CAPTF102) in the commercial culture kit and on Days 2-7 in liver infusion broth medium. Viable parasites were detectable for longer periods in liver infusion broth medium and the commercial culture kit than in Diamond's medium. Peak concentrations for isolates tended to be higher in modified Diamond's medium than in liver infusion broth medium or the commercial culture kit. Results show that these three media are suitable for the growth of all 16 T. foetus isolates from three continents and suggest that these media could be used effectively throughout the world. 相似文献
3.
为了解中国川西北牦牛肉中产志贺毒素大肠杆菌(STEC)的携带情况及stx2的亚型和特征,试验将采集的204份川西北牦牛肉样品(各25g)增菌培养后,每份挑取5个可疑菌落,采用stx1、stx2双重PCR方法检测STEC,对分离株中的stx2分型并克隆测定stx2编码区序列。结果显示,在204份样品中分离出8株STEC,平均分离率为3.9%(8/204);存在4个不同的O血清型,分别为O38(4)、O50(1)、O74(2)、O150(1);在6株含有stx2的菌株中,其中有2株为stx2a型、4株为stx2c型。结果表明牦牛源分离株氨基酸序列与人源和牛源菌株同源性较高;由stx2A、B亚基的氨基酸序列系统进化树可知,牦牛源分离株与人源、牛源菌株聚为一支,表明它们之间遗传距离相对较近,牦牛源stx2各自分布在自己的小分支中,表明牦牛源STEC stx2与人源和牛源stx2相比,尽管亲缘关系较近,但仍存在一定程度的差异。 相似文献
4.
K Shinagawa S Otake N Matsusaka S Sugii 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1992,54(3):443-446
Vacuole response in HEp-2 cells was induced with culture supernatants of Bacillus cereus strains isolated from outbreaks of vomiting- and diarrheal-type food poisoning grown in rice flour and laboratory media. High vacuole response was obtained with culture supernatants of B. cereus strains isolated from vomiting-type food poisoning grown in cooked rice suspension or on a cooked rice plate, whereas no response was obtained with those of the same strains grown in brain heart infusion and trypto-soya broth media. The vacuole activity appeared only after spore formation of B. cereus. The activity was stable to proteolytic enzymes, heating, and exposing to pH 2.0 and 11.0. Of 124 strains isolated from B. cereus food poisoning that were tested, the vacuole activity was observed by 68 of 110 (61.8%) of the strains isolated from the vomiting-type food poisoning but not by all strains (14 strains) from diarrheal-type ones. Moreover, the vacuole response in the HEp-2 cells was found to be induced by 56 of 76 (73.7%) of the serotype H-1 strains isolated from vomiting-type food poisoning. 相似文献
5.
Species of anaerobic bacteria isolated from clinical veterinary specimens were tested for susceptibility to trimethoprim-sulfonamides by the broth-disk elution technique. Three different media were used for each organism: prereduced anaerobically sterilized (PRAS) brain-heart infusion broth (BHI), thioglycollate broth, and a semidefined PRAS medium. Susceptibility results from these media were compared with those determined by interpreting the minimal inhibitory concentration obtained using an agar dilution technique. Results from broth-disk testing in semidefined medium agreed in 68.7% of the cases, in 53.7% for thioglycollate broth, and in 36.9% for BHI. The greatest deviation between techniques occurred with isolates belonging to the genus Bacteroides, followed by those of the genus Clostridium and those of the genus Fusobacterium. This deviation was directly proportional to increasing concentrations of thymidine in the BHI and thioglycollate broths but not with the semidefined medium. We conclude that the broth-disk elution method for measuring susceptibility of obligate anaerobes to trimethoprim-sulfonamides is unsuitable. 相似文献
6.
解磷根瘤菌液体培养基类型,浓度及透气条件的比较 总被引：1，自引：1，他引：0
以解磷根瘤菌SL01为供试菌株,采用液体培养研究了培养基类型和浓度以及透气状况对其生长的影响。结果表明:供试的培养基中,最适的液体种子培养基为LB和1/2 LB液体培养基,菌株的繁殖速度不仅与培养基的类型有关,更受到营养条件的限制;培养基接入解磷根瘤菌SL01并充分发酵之后,发酵液的pH值都相当程度的偏离了初始的酸碱度;根瘤菌作为好气菌,YMA液体培养基中培养12 h后,透气条件下发酵液内的活菌数达12.2×109cfu/mL,为密闭培养条件下的10.51倍,在解磷根瘤菌的发酵培养中,透气状况应作为重要的环节进行考虑。 相似文献
7.
Polyacrylamide gel electrophoresis of whole-cell preparations of Rhodococcus equi. 总被引：4，自引：1，他引：3 
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下载免费PDF全文 The whole-cell proteins of ten strains of Rhodococcus equi isolated from horses, pigs, or humans, including the type strain ATCC 6939, were examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The protein profiles of seven different capsular serotypes and the type strain were very similar when bacteria were cultured under the same conditions. Protein profiles were largely unaffected by incubation at two temperatures (30 degrees C, 37 degrees C) or times (12 h, 48 h). There were generally minor differences in protein profiles between strains grown in different media (brain heart infusion, nutrient, minca broths, tryptic soy-blood agar) with the marked exception of a prominent diffuse 17.5 kd protein which was expressed in nutrient broth. This protein was not produced by the type strain and was lost on repeated passage in vitro (50th, 100th passage) in two of three other strains examined. 相似文献
8.
As part of the basic characterization of Ornithobacterium rhinotracheale, the minimal inhibitory concentrations of 10 antimicrobial drugs were determined for reference strains and Mexican isolates by a broth microdilution method. For optimal growth of the organisms, a supplemented brain-heart infusion broth was used. The susceptibility of O. rhinotracheale to amoxicillin, enrofloxacin, and oxytetracycline was variable. However, consistent higher minimal inhibitory concentrations values were obtained for gentamicin, fosfomycin, trimethoprim, sulfamethazine, sulfamerazine, sulfaquinoxaline, and sulfachloropyridazine. Obtained results among Mexican isolates indicate a marked antimicrobial drug resistance trend. 相似文献
9.
The production of M-protein antigen of Streptococcus equi was studied during in vitro growth in equine blood and in various media. Of 11 S equi strains studied, seven which had initially possessed 0.04 mg or less M-protein per 10 mg of streptococcal cell extract showed an increase in M-protein content after successive culture in heparinised horse blood. Maximum proliferation occurred in Todd-Hewitt (TH) medium with added 0.2 per cent w/v glucose when compared with TH medium alone or TH medium with 2 per cent w/v sucrose, starch, neopeptone or normal horse serum. The M-protein of these strains did not change after the addition of either neopeptone or normal horse serum to TH medium but declined with the addition of sugars. In experiments involving phagocytosis of S equi by equine polymorphs, the percentage of polymorphs which engulfed cocci was higher with a capsule-deficient strain (69.0 +/- 11.6 per cent) than with five typical encapsulated strains (21.1 +/- 7.0 per cent to 30.9 +/- 13.3 per cent). Phagocytosis of five typical strains was greater after growth in a trypsin-containing medium than in medium devoid of trypsin. Trypsin-grown cells took longer to kill mice than did normal cells. It was concluded that M-protein was one of the factors involved in the virulence of S equi. 相似文献
10.
通过体外α-葡萄糖苷酶抑制试验和糖尿病模型小鼠动物试验,检验桑树内生菌菌株Stenotrophomonas maltophilia Y1和Micrococcus luteus Y2发酵产物的降血糖作用。结果表明,Y1和Y2菌株发酵液提取物对α-葡萄糖苷酶均具有显著的体外抑制作用,并呈现一定的剂量依赖性,半数抑制浓度(IC50)分别为2 307.56 mg/L和111.58 mg/L,在试验最大剂量下的抑制率分别达到59.17%和69.21%,均高于对照药物拜糖平(抑制率为53.06%)。动物试验表明,饲喂Y1菌株发酵液浓缩产物的糖尿病模型小鼠的空腹血糖(FBG)值和餐后血糖(PBG)值均低于模型组,分别存在显著和极显著差异,而对正常小鼠的血糖无明显影响,也未表现出毒副作用;调查饲喂发酵产物后糖尿病模型小鼠的体质量、进食量、饮水量、排尿量等指标,均得到明显改善。试验结果显示,2株桑树内生菌菌株的发酵液提取物均表现出与1-脱氧野尻霉素(DNJ)相同的体外抑制α-葡萄糖苷酶活性的作用,Y1菌株发酵产物表现出与DNJ相同的降低糖尿病模型小鼠血糖及改善糖尿病症状的作用特点。 相似文献
11.
The type strain WVU 1853 and field strains SG, N26 and A642 of Mycoplasma synoviae were examined for their requirement for nicotinamide-adenine dinucleotide (NAD) for in vitro growth. All the strains grew and could be repeatedly passaged in Frey broth medium supplemented with filter-sterilised NAD. In modified Frey broth medium from which NAD was omitted and broth medium for which the supplements including yeast extract and NAD were autoclaved, only strains N26 and A642 could be grown and passaged. The growth curves of strain N26 determined in broth media with and without NAD were similar. These results indicated that there are differences in NAD-requirement for in vitro growth among strains of M synoviae. 相似文献
12.
本试验旨在从实验室保藏菌株中筛选出可用于羽毛降解的菌株并优化其培养条件,应用于羽毛的生物降解。以酪蛋白培养基进行菌株初筛,以羽毛为唯一有机营养源制作的羽毛培养基进行菌株复筛;通过细菌形态学观察和16S rDNA序列测定进行菌株鉴定,并对所筛菌株在羽毛培养基中的培养条件进行优化研究。结果表明：1)通过酪蛋白培养基筛选出8株能产生降解圈且透明度高的菌株。2)通过单根羽毛培养基和羽毛降解液指标测定联合方法筛选出1株可3 d降解羽毛、产可溶性蛋白的菌株NLG1。3)经鉴定并命名为贝莱斯芽孢杆菌NLG1(Bacillus velezensis NLG1)。4)菌株NLG1培养优化条件为羽毛底物浓度2.5%(m/v),初始pH 10,接种活菌数109CFU/mL,温度27℃,外加碳源为可溶性淀粉,发酵时间3 d。5)优化条件下降解羽毛,测得发酵液中可溶性蛋白含量为(32.76±0.07)μg/mL,水解氨基酸总量由27.41 mg/g提升到112.18 mg/g,测得游离氨基酸及衍生物的种类由25种增加至31种(增加的氨基酸为胱氨酸、苯丙氨酸、天冬酰胺、α-氨基丁酸、β-氨基异丁酸、β-丙氨酸),... 相似文献
13.
Growth of Mycobacterium paratuberculosis in radiometric, Middlebrook and egg-based media 总被引：2，自引：0，他引：2
The ability of BACTEC radiometric 7H12 broth, Middlebrook 7H10 Tween broth, Middlebrook 7H10 agar, and Herrold's egg-yolk medium to provide early detection of Mycobacterium paratuberculosis was evaluated. The minimum detection times in days for the various media were: 7H12, 9; 7H10 agar, 23 (plate), 28 (slant); 7H10 Tween broth; 27; and Herrold's egg-yolk medium, 43 (plate), 49 (slant). The radiometric broths provided the earliest detection of M. paratuberculosis, and 3625 organisms ml-1 were required to produce a positive, radiometric growth-index reading. Of the non-radiometric plate and slant media evaluated, microscope examination of the translucent 7H10 agar plate resulted in the earliest detection and highest mean colony counts (387) as compared with Herrold's egg-yolk agar plate (208). Similar results were noted for 7H10 and Herrold's egg-yolk agar slants; however, accurate colony counts could not be determined because of confluent growth. All media were supplemented with 2 micrograms ml-1 of mycobactin J and excess amounts of this supplement inhibited the growth of M. paratuberculosis in radiometric 7H12 media. 相似文献
14.
Effects of temperature on growth of three strains of Bordetella avium were determined in young turkeys and in vitro. Colonization of the tracheal mucosa by two virulent strains of B. avium was significantly greater in cold-stressed turkeys than in heat-stressed turkeys. The avirulent vaccine strain, ART-VAX, colonized tracheas of cold-stressed turkeys to a limited extent but failed to colonize heat-stressed turkeys. Growth rates of the three B. avium strains were determined in brain-heart infusion broth at 30, 35, 40, and 45 C. All three strains grew best at 35 C but were killed by 45 C. Compared with virulent strains, ART-VAX grew markedly less at all temperatures, and most cultures of ART-VAX grew at 40 C only after a variable period of declining numbers of viable bacteria. This study indicates that temperature affects growth of B. avium in vivo and in vitro and that growth of the ART-VAX strain is fundamentally different from growth of virulent strains. 相似文献
15.
De Meerschman F Rettigner C Focant C Boreux R Pinset C Leclipteux T Losson B 《Veterinary research》2002,33(2):159-168
Neospora caninum and Toxoplasma gondii are cyst-forming coccidian parasites of human and veterinary clinical relevance. In vitro cultivation of the protozoans using Vero cells is usually performed in order to produce antigenic materials. Quantitative and qualitative comparisons of Vero cells grown in RPMI medium supplemented either with foetal calf serum (FCS), horse serum (HS) or a specific serum-free additive (DefCell) were performed. A serum-free cell culture system used to propagate N. caninum (NC-1 isolate) and T. gondii tachyzoites (Rh stain) were compared with the other two cell culture systems. FCS supplemented media was found to be more effective than the others in promoting Vero cells and N. caninum tachyzoites. However, it was found unable to support adequate T. gondii tachyzoite proliferation. Vero cells, T. gondii and N. caninum tachyzoite production gave similar growth patterns with either HS or DefCell supplemented media. Defcell was considered as a good alternative to supplement culture medium. 相似文献
16.
Calderaro A Bommezzadri S Piccolo G Zuelli C Dettori G Chezzi C 《Veterinary microbiology》2005,105(3-4):229-234
The aim of this study was to compare and evaluate the time required to isolate Brachyspira hyodysenteriae and Brachyspira pilosicoli from porcine faeces. This was done using previously described selective media (spectinomycin) S400, (colistin, vancomycin and spectinomycin) CVS and (spectinomycin, vancomycin, colistin, spiramycin and rifampin with swine faecal extract) BJ, compared with the method based on blood agar modified medium, with spectinomycin and rifampin (BAM-SR), including a pre-treatment step. Fourteen spirochaetal strains were obtained in pure cultures after 5 days (48 h in BAM-SR primary plate and three passages every 24 h in brain heart infusion (BHI) without antibiotics) pre-treating simulated samples in brain heart infusion broth with spectinomycin (400 microg/ml) and rifampin (15 microg/ml), before streaking on the selective BAM-SR medium. Spirochaetes from samples in S400, CVS and BJ, with and without pre-treatment, were obtained in pure cultures only after repeatedly transferring on plates of the same selective medium requiring 15-18 days according to the strain. BAM-SR used after the pre-treatment step showed a detection limit ranging from 3.5 x 10(2) to 6.7 x 10(7) cells/g faeces and was the only method able to support the growth of spirochaetes after 48 h. 相似文献
17.
A co-culture assay for isolation of Borrelia burgdorferi sensu lato (sl.) from naturally infected ticks and dogs suspected of Lyme borreliosis (LB) was evaluated using buffalo-green-monkey (BGM) cells as the mammalian component. Four different media were tested for their ability to provide sufficient growth conditions for spirochetes and BGM cells. A total of 176 Ixodes ricinus ticks and 268 specimens from 98 dogs were used to compare cell-free culture with the BGM co-culture. A 1:1 mixture of Barbour-Stoenner-Kelly medium (BSK) and Eagle's minimum essential medium (EMEM) supported the growth of the two test strains, B. burgdorferi sensu stricto B31 and B. valaisiana VS116 to the same extent as BSK medium and the growth as well as the viability of BGM cells in this medium were the same as in EMEM. Using the 1:1 mixture of BSK and EMEM, borrelial growth measured in co-culture with BGM cells did not differ significantly from corresponding values obtained in cell-free cultures. In cell-free culture the isolation rate of B. burgdorferi sl. from ticks was significantly higher in BSK/EMEM 1:1 than in BSK medium (P < 0.01). Co-culture with BGM cells had no significant influence on the isolation rate of borreliae from ticks. However, a significant amount of isolates were obtained by one of the procedures only. Analysing canine specimens accordingly, spirochetes were grown from the blood of one dog after four weeks in BGM cell co-culture. The isolate was classified as B. afzelii by PCR-coupled restriction fragment length polymorphism analysis. 相似文献
18.
SU Zhanqiang WANG Dong ZHANG Jinyu ZHANG Yi LIU Yingyu MA Kaiqi GAO Jiaojiao SUN Xue YAO Gang 《中国畜牧兽医》2018,45(1):212-218
To study the impact of season to the distribution of bovine E.coli O157:H7,samples of anus swabs (399),feces (68),water (29) and feed (43) were collected in the spring, summer,autumn and winter from A,B and C farms of Xinjiang. After enrichment by EC broth, SMAC and MUG selective culture were then performed. Finally,PCR was used for identification and virulence gene detection of isolated strains. A total of 5 E.coli O157:H7 strains were isolated from 539 samples from three farms (0.93%,5/539), 2 of them were from spring (1.44%,2/139),1 from autumn(0.56%,1/180),2 from winter (1.38%,2/145) and no strains were isolated from summer samples. One strain were isolated from anus swab samples in farm B (0.69%,1/145) and one were isolated from anus swabs (0.66%,1/152) and three strains were isolated from feed samples in farm C (20.00%,3/15),and no target strains were isolated from water samples. The distribution of bovine E. coli O157:H7 had obvious seasonal characteristics.One E.coli O157:H7 strain of farm B was isolated from autumn and four of farm C were from isolated spring (2 strain) and winter (2 strain),and the isolation rate of E. coli O157:H7 in spring and winter were higher than that in summer and autumn. In conclusion,under the special climate characteristics and feeding mode in Xinjiang,to prevent and control the spreading of E. coli O157:H7 of cattle,we must pay great attention on hygiene management of pens at cold season, specially avoiding the feed contaminated by feces. 相似文献
19.
Audria M. Buchanan 《Veterinary microbiology》1982,7(6):587-605
The present study has shown that a catalase-positive Actinomyces-like organism, UCD 19-480, failed to grom from its clinical fluid origin as normal colonies on three types of L-form media, although all three media were later shown to support it from a blood agar plate source. The growth of 19-480 from a fresh sulfur granule into a droplet of modified Barile Yaguchi Eveland (BYE) L-form broth was studied in a sealed slide incubated at 25–28°C for 11 weeks. Within 7 days, very long, bizarre branched filaments were detected near the crushed granule mass. Microcolonies were present at 5 weeks with radial filaments extending 0.5 cm from a core. The core consisted of pleomorphic vesicles arranged in a ring. Crystals appeared within and around the granule, but not elsewhere. After repeated passage on blood agar plates, 19-480 and a second, non-granule strain of recent origin, 339-180, were inoculated into BYE L-form broths as well as broths of new design. Unique membranous structures exhibiting actinomycete morphology were detected at 3 weeks in the 19-480 trypticase soya cultures containing putrescine and at 9 weeks in BYE L-form broths. They were restricted to the deep portions of certain broth types at predictable times and were not present in incubated controls. They fluoresced intensely with acridine orange and were weakly Gram-positive. ‘Pseudocolony’-like formations were detected at 8 weeks in the 3% NaCl + 5% sucrose BYE broth culture of UCD 19-480. It is suggested that the absence of normal colonies of these agents of L-form plate cultures of clinical fluids is due to growth of spreading variants unique to the actinomycete group. 相似文献
20.
In Germany treatment of swine dysentery is hampered by Brachyspira hyodysenteriae strains showing elevated MIC values to the few antibiotics licensed. Therefore, susceptibility testing of clinical isolates is an important service to the swine practitioner. This study compares the established agar dilution procedure for antimicrobial susceptibility testing of this fastidious anaerobe to the broth microdilution test newly developed [Anim. Health Res. 2 (2001) 59; Vet. Microbiol. 84 (2002) 123; J. Clin. Microbiol. 41 (2003) 2596]. A total of 221 isolates were examined twice with either test procedure using tiamulin and valnemulin as antibiotics. Both methods gave reproducible results, and the MIC values for the reference strains B. hyodysenteriae B204 and Staphylococcus aureus ATCC 29213 corresponded to previously published data. However, the results for individual strains differed significantly for both tests (P < 0.001) with MIC values being on average one dilution step lower in the broth dilution method. The 221 strains used for comparing test procedures were isolated between 1989 and 2001. An additional 102 strains isolated in 2002 were tested only with the broth dilution procedure. A significant rise in the average MIC value for both pleuromutilins could be demonstrated when comparing earlier isolates to those from 2000 to 2001 (P < 0.05), while in 2002 the average MIC significantly decreased when compared to the value in 2000 (P < 0.05). However, strains with MIC values for tiamulin as high as 8 microg/ml (broth dilution) could still be isolated. 相似文献