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1.
Yamamoto A Iwata A Saitoh T Tuchiya K Kanai T Tsujimoto H Hasegawa A Ishihama A Ueda S 《Veterinary immunology and immunopathology》2002,90(3-4):169-177
Feline granulocyte colony-stimulating factor (G-CSF) with an N-terminal histidine hexamer tag was expressed as inclusion bodies in E. coli. The G-CSF solubilized in 6 M guanidine solution was absorbed onto a Ni-NTA column and, after washing with decreasing concentrations of guanidine, eluted with imidazole in a soluble and apparently pure form. The activity of the recombinant feline G-CSF was 3×106 U/mg protein, as assayed by its stimulatory effect on NFS-60 cell proliferation. When a low level of purified feline G-CSF was administered once a day for two successive days to cats, the number of neutrophil increased 4-fold while the levels of other blood cell types remained virtually unchanged. Daily administration of G-CSF for a total of 11 days led to a more than 10-fold increase in neutrophils, an 8-fold increase in the number of monocytes and 2-fold increase in lymphocytes. No severe side effects or antibody production was observed in cats after administration of G-CSF. 相似文献
2.
Akira Yamamoto Akira Iwata Toshiki Saito Fumiko Watanabe Susumu Ueda 《Veterinary immunology and immunopathology》2009,130(3-4):221-225
Canine granulocyte colony-stimulating factor (cG-CSF) with modification of cysteine at position 17 to serine was expressed in Brevibacillus choshinensis HPD31. cG-CSF secreted into the culture medium was purified by ammonium sulfate precipitation and consecutive column chromatography, using butyl sepharose and DEAE sepharose. Biological activity of the recombinant cG-CSF was 8.0 × 106 U/mg protein, as determined by its stimulatory effect on NFS-60 cell proliferation. Purified cG-CSF was subcutaneously administered once a day for two successive days to dogs (1, 5, 25, or 125 μg). Neutrophil count increased the following day in all dogs except those administered the lowest dose (1 μg). No severe side effects were observed in dogs after administration of cG-CSF. 相似文献
3.
Tetracyclines and bacitracin are used extensively as a growth promotant and a therapeutic, respectively in livestock. In this study, we test whether there is an interaction between bacitracin and tetracycline that can select for an increase in microbial tetracycline sensitivity, using Escherichia coli and Salmonella as a model. We found via in vitro studies that bacitracin at sublethal concentrations preferentially selects for outgrowth of tetracycline-sensitive bacteria from among a population of tetracycline-resistant and tetracycline-sensitive E. coli and Salmonella strains. Most of the bacterial strains employed in our study were shown by PCR to possess the tetracycline resistance gene tet(A) or tet(C), either of whose activity is associated with tetracycline efflux. Bacitracin could be substituted for the lipophylic chelator, fusaric acid, used as a positive selective agent for tetracycline sensitivity. Together, these observations suggest that bacitracin-mediated selection for tetracycline sensitivity alters the function of tetracycline efflux proteins. Using bacitracin and tetracycline simultaneously may improve the effectiveness of the antibiotics, while decreasing the risk of selecting for a population of tetracycline-resistant microorganisms. 相似文献
4.
P Maisi T Mattila M Sandholm 《Comparative immunology, microbiology and infectious diseases》1983,6(4):273-280
Growth of Escherichia coli was followed in sow whey by a turbidometric technique. Heme-iron clearly promoted bacterial growth, therefore free heme would be dangerous in small intestine by enhancing bacterial growth: duodenal brush border membrane (BBM) was seen to bind heme-iron, thus abolishing the growth promoting effect of this form of iron. Scatchard analysis of heme-binding onto BBM showed that heme became bound by a specific mechanism (KD ≈ 10-7 M) as well as non-specifically. The absorption system for iron within the enterocytes should be considered an important antibacterial system within the gut. 相似文献
5.
Bashar W. Shaheen Chengming Wang Calvin M. Johnson Bernhard Kaltenboeck Dawn M. Boothe 《Veterinary microbiology》2009,139(3-4):379-385
Fluoroquinolones are used to treat infections caused by Escherichia coli in canine and feline veterinary patients, particularly those infecting the urinary tract. The gyrA gene is a primary target causing fluoroquinolone resistance in Gram negative coliforms, with mutations in codons 83 and 87 generally associated with high-level of resistance E. coli clinical isolates. We have developed a fluorescence resonance energy transfer (FRET) quantitative PCR to identify enrofloxacin-resistance in clinical E. coli isolates that carry mutations in codons 83 and 87 of gyrA. This real-time quantitative PCR assay is rapid, economical, and sensitive compared with cultured antimicrobial susceptibility testing. The assay identified as few as four genome copies per reaction from culture and 19 genome copies in urine. For the 70 isolates tested, the sensitivity was 87.5% (95% CI = 75–95.3%) (n = 42/48), specificity was 100% (95% CI = 87.3–100%) (n = 22/22), whereas accuracy was 91.4% (95% CI = 82.3–97%) (n = 64/70). Furthermore, we were able to accurately differentiate between the wild type and mutants E. coli directly from infected canine urine samples (n = 5) within 2 h. These results were confirmed by sequence alignments of the PCR products and comparison with the susceptibility testing. The FRET-PCR assay appears to have promising clinical application as an early diagnostic tool for rapid and sensitive detection and differentiation of the level of fluoroquinolone resistance among clinical E. coli isolates that may facilitate design of the dosing regimen. 相似文献
6.
Shota Kono Tomohiko Kazama Koichiro Kano Kayoko Harada Masami Uechi Taro Matsumoto 《Veterinary journal (London, England : 1997)》2014,199(1):88-96
It has been reported that mature adipocyte-derived dedifferentiated fat (DFAT) cells show multilineage differentiation potential similar to that observed in mesenchymal stem cells. Since DFAT cells can be prepared from a small quantity of adipose tissue, they could facilitate cell-based therapies in small companion animals such as cats. The present study examined whether multipotent DFAT cells can be generated from feline adipose tissue, and the properties of DFAT cells were compared with those of adipose-derived stem cells (ASCs). DFAT cells and ASCs were prepared from the floating mature adipocyte fraction and the stromal vascular fraction, respectively, of collagenase-digested feline omental adipose tissue. Both cell types were evaluated for growth kinetics, colony-forming unit fibroblast (CFU-F) frequency, immunophenotypic properties, and multilineage differentiation potential.DFAT cells and ASCs could be generated from approximately 1 g of adipose tissue and were grown and subcultured on laminin-coated dishes. The frequency of CFU-Fs in DFAT cells (35.8%) was significantly higher than that in ASCs (20.8%) at passage 1 (P1). DFAT cells and ASCs displayed similar immunophenotypes (CD44+, CD90+, CD105+, CD14?, CD34? and CD45?). Alpha-smooth muscle actin-positive cells were readily detected in ASCs (15.2 ± 7.2%) but were rare in DFAT cells (2.2 ± 3.2%) at P1. Both cell types exhibited adipogenic, osteogenic, chondrogenic, and smooth muscle cell differentiation potential in vitro. In conclusion, feline DFAT cells exhibited similar properties to ASCs but displayed higher CFU-F frequency and greater homogeneity. DFAT cells, like ASCs, may be an attractive source for cell-based therapies in cats. 相似文献
7.
In vitro methods were used to test the hypothesis that Escherichia coli from bovine mastitis are essentially no different from isolates from bovine feces. Fifty E. coli isolates from bovine mastitic milk, 50 from feces of mastitic cows and 50 from feces of healthy cows were compared with respect to biochemical properties and certain potential virulence factors. There were no significant differences among the groups in tests for biotype; production of colicins, colicin V, or Vero cell cytotocity; and growth in 90% gnotobiotic calf serum or 90% normal milk whey. Resistance to killing in 90% gnotobiotic calf serum varied from 66 to 84%. Most isolates grew in normal whey: the percentage in a group varied from 86 to 96. Mastitic milk isolates were significantly different from the fecal isolates in adonitol fermentation (P0.006), production of aerobactin (P0.026), and ability to grow in 90% mastitic whey (P0.00004). However, only 40% of mastitis E. coli fermented adonitol and only 20% produced aerobactin. Ninety-six percent of mastitic milk E. coli grew in mastitic whey, whereas 64% and 60%, respectively, of mastitic fecal and normal fecal isolates grew in this medium. It is concluded that none of the properties that were investigated constitute potential virulence factors or markers for ability to induce mastitis; the data are consistent with the hypothesis that mastitic E. coli are simply opportunistic pathogens. 相似文献
8.
A. Fernandez C. Lara R. Puyuelo J. Gomez J.J. Ramos A. Loste M.C. Marca M.T. Verde 《Research in veterinary science》1998,65(3):201
Seventy-five 25-day-old broilers were divided into three groups: group I unmedicated and challenged with E coli 078:K80; group F infected and treated with 150 ppm of phosphomycin in their drinking water, and group C acted as a control. Their weights, feed intake, clinical signs, macroscopic lesions, E coli reisolation, and serum biochemistry were compared. Group F showed fewer symptoms and gross lesions than those from group I while the average daily gain, bodyweight, and feed intake were similar to the control group. E coli was reisolated in 32 per cent of the livers and spleens from group I, compared with 4 per cent of liver and 8 per cent of spleens from group F. There was an increase in the levels of total protein and globulins in group I but not in group F. These results provide evidence of the therapeutic efficacy of phosphomycin in the control of an experimental E coli infection in broiler chickens. 相似文献
9.
Daniela Costa Laura Vinu Patricia Poeta Ana Cludia Coelho Manuela Matos Yolanda Senz Sergio Somalo Myriam Zarazaga Jorge Rodrigues Carmen Torres 《Veterinary microbiology》2009,138(3-4):339-344
Seventy-six faecal samples were obtained from broilers at slaughterhouse level in Portugal. Samples were inoculated on cefotaxime-supplemented Levine agar plates. Cefotaxime-resistant Escherichia coli isolates were recovered from 32 samples (42.1%), obtaining a total of 34 E. coli isolates (one or two isolates per sample). Susceptibility to 16 antibiotics was studied by disk diffusion method, and 85% of the isolates presented a phenotype of multi-resistance that included antimicrobial agents of at least four different families. Extended-spectrum-beta-lactamases (ESBL) of the TEM and CTX-M groups were detected in 31 ESBL-positive E. coli isolates. Twenty-six isolates harboured the blaTEM-52 gene and two of them also harboured blaTEM-1b. The blaCTX-M-14 gene was identified in three isolates (in association with blaTEM-1b in one of them), and blaCTX-M-32 was demonstrated in two additional isolates. Three of the 34 cefotaxime-resistant isolates (9%) did not produce ESBLs, and two of them presented mutations at positions −42 (C → T), −18 (G → A), −1 (C → T), and +58(C → T) of the promoter/attenuator region of ampC gene. tet(A) and/or tet(B) genes were detected in all 34 tetracycline-resistant isolates, aadA in all 26 streptomycin-resistant isolates; cmlA in 3 of 6 chloramphenicol-resistant isolates, and aac(3)-II or aac(3)-I + aac(3)-IV genes in all 4 gentamicin-resistant isolates. Different combinations of sul1, sul2 and sul3 genes were demonstrated among the 22 trimethoprim–sulfamethoxazole-resistant isolates. Amino acid changes in GyrA and ParC proteins were identified in all 18 ciprofloxacin-resistant isolates. The results of this study indicate that the intestinal tract of healthy poultry is a reservoir of ESBL-positive E. coli isolates. 相似文献
10.
Jacek Osek Peter Gallien Dieter Protz 《Comparative immunology, microbiology and infectious diseases》2000,23(4)
Fecal samples from 67 3–5-months-old calves with diarrhea were screened for the presence of shiga toxin-producing Escherichia coli (STEC). Several accessory virulence factors genes were also tested. Among 192 E.coli isolates tested, 15 (7.6%) were found to harbour the shiga toxin 1 or 2 (stx1 or stx2) genes. The stx2-carrying samples were further subtyped by PCR for the stx2c, stx2d, and stx2e toxin variants. It was shown that stx2-positive bacteria mainly possessed the stx2c shiga toxin type gene. The enterohemolysin (hlyA) and intimin (eae) genes were found in seven (46.7%) STEC strains whereas the cytotoxic necrotizin factor 1 and 2 or the P fimbrial genes were detected in two isolates only. This study confirmed that calves are a reservoir of STEC strains (with all pathogenicity genes) that may be virulent for humans. 相似文献
11.
B.A. Hales J.N. Fletcher G. Ridha R.M. Batt C.A. Hart J.R. Saunders 《Research in veterinary science》1991,50(3):9-357
Seventeen bovine and 56 porcine Escherichia coli isolates from cases of diarrhoea and from healthy animals were examined for DNA sequences homologous to the genes for verocytotoxins (VT), enterotoxins and human enterohaemorrhagic E coli/enteropathogenic E coli (EHEC EPEC) sequences. VT-1 was the most common toxin among the bovine isolates and VT-2 the most common in the porcine isolates. No isolates had homologous sequences to enteropathogenic adherence factor, but 71.2 per cent hybridised to the DNA probe encoding specific EHEC sequences, and 95.9 per cent showed homology with a 23 kb DNA fragment common to EHEC and EPEC plasmids. 相似文献
12.
Martina Becker Andreas Moritz Urs Giger 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2008,37(4):373-384
Background: A CBC is an integral part of the assessment of health and disease in companion animals. While in the past newer technologies for CBC analysis were limited to large clinical pathology laboratories, several smaller and affordable automated hematology analyzers have been developed for in‐clinic use. Objectives: The purpose of this study was to compare CBC results generated by 7 in‐clinic laser‐ and impedance‐based hematology instruments and 2 commercial laboratory analyzers. Methods: Over a 3‐month period, fresh EDTA‐anticoagulated blood samples from healthy and diseased dogs (n=260) and cats (n=110) were analyzed on the LaserCyte, ForCyte, MS45, Heska CBC, Scil Vet ABC, VetScan HMT, QBC Vet Autoread, CELL‐DYN 3500, and ADVIA 120 analyzers. Results were compared by regression correlation (linear, Deming, Passing‐Bablok) and Bland–Altman bias plots using the ADVIA as the criterion standard for all analytes except HCT, which was compared with manual PCV. Precision, linearity, and carryover also were evaluated. Results: For most analytes, the in‐clinic analyzers and the CELL‐DYN performed similarly and correlated well with the ADVIA. The biases ranged from ?0.6 to 2.4 × 109/L for WBC count, 0 to 0.9 × 1012/L for RBC count, ?1.5 to 0.7 g/dL for hemoglobin concentration, ?4.3 to 8.3 fL for MCV, and ?69.3 to 77.2 × 109/L for platelet count. Compared with PCV, the HCT on most analyzers had a bias from 0.1% to 7.2%. Canine reticulocyte counts on the LaserCyte and ForCyte correlated but had a negative bias compared with those on the ADVIA. Precision, linearity, and carryover results were excellent for most analyzers. Conclusions: Total WBC and RBC counts were acceptable on all in‐clinic hematology instruments studied, with limitations for some RBC parameters and platelet counts. Together with evaluation of a blood film, these in‐clinic instruments can provide useful information on canine and feline patients in veterinary practices. 相似文献
13.
The purpose of this study was to measure the net absorption of fluid, sodium, potassium and chloride in the small intestine of weaned pigs and of their unweaned littermates and to correlate these values with villus height and crypt depth. Five pairs of segments of the small intestine were prepared in each of 80 pigs; the cranial segment of each pair was injected with an enterotoxigenic strain of Escherichia coli and the caudal segment with a control solution. Net absorption was measured on the day of weaning and four, seven, 11 and 14 days after weaning. In unweaned pigs the net absorption of fluid, potassium and chloride did not vary with time. In weaned pigs the net absorption of fluid in the control segments was significantly less on days 4, 7 and 14 after weaning and of sodium and chloride on days 4 and 7 than in unweaned littermates. In infected segments of weaned pigs the net absorption of fluid was significantly less than in unweaned littermates on day 11 and 14, of sodium and potassium on day 11 and of chloride on days 4 and 11 after weaning. Net absorption was negatively correlated with villus height but only in the infected segments of weaned pigs; no other correlations were found. It was concluded that after weaning the net absorption of fluid and electrolytes in the small intestine of pigs is temporarily decreased, a condition that may initiate diarrhoea. 相似文献
14.
15.
Konrad A. Dziaba Gabriele Lambrecht Klaus Petzoldt 《Comparative immunology, microbiology and infectious diseases》1985,8(3-4):267-272
The local and systemic immune response to a formolized E. coli oral vaccine was investigated in 13 gnotobiotic piglets. Beginning at ten days of age animals received a daily dose of 1010 or 1011 bacteria, on ten consecutive days. Intestinal loop tests with one animal of each group on day 26 showed protection which was more pronounced in the animal dosed 1010 bacteria compared with the other immunized piglet. Immunoglobulin class-specific antibodies to O and K antigens were determined by ELISA technique. In serum no IgG or IgA antibodies were found, whereas IgM-anti O149 antibodies in both immunized groups reached their highest level at day 4 of dosing and decreased thereafter. IgM-anti K88 antibodies were first detected at day 10 of dosing. Both immunized groups had comparable serum levels at days 20 and 30. Also in gut secretion the IgM antibody response was predominant, and higher levels were found in the 1010 group than in the 1011 group. IgG and IgA antibody response were also detected in secretion. 相似文献
16.
牛肉源大肠杆菌的耐药性检测及相关耐药基因分布 总被引:3,自引:3,他引:3
为了解牛肉源大肠杆菌的耐药性并检测其相关耐药基因分布,本研究选取了117株牛肉源大肠杆菌,经药敏纸片法对11种抗菌药物进行了药敏检测,并根据耐药表型利用普通和(或)多重PCR技术对耐四环素菌中tet(A)、tet(B)和tet(C)基因,耐氨苄西林菌中blaTEM1、blaPSE1和blaOXA1基因,耐链霉素菌中strA-strB、aadA1基因,耐磺胺类药菌中sul1、sul2和sul3基因进行了调查分析。结果显示,117株大肠杆菌对四环素、氨苄西林、链霉素和磺胺异恶唑的耐药率较高,分别为89%、42%、38%和22%。tet(A)基因是所有四环素耐药基因中最为流行的一种基因(55%);在检测的3个β-内酰胺类药物耐药基因中,最流行的为blaTEM1基因(73%);链霉素的耐药性主要由strA-strB基因(38%)编码;sul2基因在耐磺胺异恶唑菌中的检出率最高(77%)。结果表明本次分离的牛肉源大肠杆菌耐药非常严重,进一步肯定了肉牛业生产中抗菌药的使用对大肠杆菌耐药性的产生和发展所发挥的重要作用,提示食品动物养殖应严格控制饲料中抗菌药的滥用。 相似文献
17.
Virulence gene and antibiotic resistance profile of Shiga-toxin-producing Escherichia coli prevalent in captive yaks (Poephagus grunniens) 总被引:2,自引:0,他引:2
Samiran Bandyopadhyay Tapas K. Biswas Devasis Sasmal Monoj K. Ghosh Tapan K. Dutta Suresh C. Das Debasis Bhattacharya Asit K. Bera Subhasis Bandyopadhyay Sumanta De Diganta Pan 《Veterinary microbiology》2009,138(3-4):403-404
18.
Emad E. Younis Ashraf M. Ahmed Sabry A. El-Khodery Salama A. Osman Yasser F.I. El-Naker 《Research in veterinary science》2009,87(3):373-379
The aim of the present study was to carry out molecular epidemiological investigation on enterotoxigenic Escherichia coli (ETEC) K99 and Salmonella spp. in diarrheic neonatal calves. Fecal samples were obtained from 220 diarrheic calves at 9 farms related to four governorates in central and northern Egypt. E. coli and Salmonella spp. isolates were examined for E. coli K99 and Salmonella spp. using PCR. ETEC K99 was recovered from 20 (10.36 %) out of 193 isolates, whereas Salmonella spp. was recovered from nine calves (4.09%).Multivariable logistic regression was used to evaluate the risk factors associated with both infections. ETEC K99 was significantly affected by age (P < 0.01; OR: 1.812; CI 95%: 0.566–1.769), colostrum feeding practice (P < 0.01; OR: 5.525; CI 95%: 2.025–15.076), rotavirus infection (P < 0.001; OR: 2.220; CI 95%: 0.273–1.251), vaccination of pregnant dams with combined vaccine against rotavirus, coronavirus and E. coli (K99) (P < 0.001; OR: 4.753; CI 95%: 2.124–10.641), and vitamin E and selenium administration to the pregnant dam (P < 0.01; OR: 3.933; CI 95%: 0.703–1.248).Infection with Salmonella spp. was found to be significantly affected by the animal age (P < 0.05; OR: 0.376; CI 95%: 0.511–1.369), Hygiene (P < 0.05; OR: 0.628; CI 95%: 1.729–5.612), and region (P < 0. 01; OR: 0.970; CI 95%: 0.841–1.624).The results of the present study indicate the importance of PCR as rapid, effective and reliable tool for screening of ETEC and Salmonella spp. when confronted with cases of undifferentiated calf diarrhea. Moreover, identification of the risk factors associated with the spreading of bacteria causing diarrhea may be helpful for construction of suitable methods for prevention and control. 相似文献
19.
D. Ciosek M. Truszczyski 《Comparative immunology, microbiology and infectious diseases》1982,5(4):405-412
Serotypes of E. coli strains isolated from piglets, which died with symptoms of diarrhea in 9 swine industrial farms, were determined. Large numbers of serotypes (from 16 to 27) in individual farms were detected. The sets of serotypes from 9 investigated farms differed among each other significantly, depending on the farm and time of examination. It was found that more than one serotype of E. coli may exist in the pig body and contribute to the development of disease. The predominant serotypes, i.e. those comprising more than 10% of serologically determined strains, were found to exist in 6 of the investigated farms and not in the remaining ones. Among the predominant serotypes, particularly important seem to be strains with K88 antigen. For prophylaxis of piglet colibacteriosis in industrial farms in Poland two vaccines for sows are recommended: one containing the K88 antigen only and the other the following serotypes: 0149:K91,K88; 020:K57; 020:K83; 0157:K88; 01:K1; 0136:K78; 024:K?; 078:K80 and 0118:K? Strains belonging to these serotypes were the most prevalent in our strain collection. 相似文献
20.
The aim of this study was to quantify and compare interferon-γ (IFN-γ) concentrations in the serum of clinically normal cats infected with feline coronavirus (FCoV) with its concentration in the sera and effusions of cats with feline infectious peritonitis (FIP), a disease associated with infection with a mutated form of FCoV.Clinically normal FCoV-infected cats living in catteries with a high prevalence of FIP had the highest serum IFN-γ concentrations. The serum concentration of IFN-γ was not significantly different in cats with FIP compared with clinically normal FCoV-infected animals living in catteries with a low prevalence of the disease. Moreover, the concentration of IFN-γ was significantly higher in the effusions than in the serum of cats with FIP, probably due to IFN-γ production within lesions. These findings support the hypothesis that there is a strong, ‘systemic’ cell mediated immune response in clinically normal, FCoV-infected cats and that a similar process, albeit at a tissue level, is involved in the pathogenesis of FIP. 相似文献