首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献，搜索用时 546 毫秒
1.
Haemophilus parasuis (H. parasuis) is a swine pathogen responsible for the Glässer’s disease. In order to understand the pathogenesis of the H. parasuis infection, the gnd gene encoding a cell surface protein, 6-phosphogluconate-dehydrogenase (6PGD) of H. parasuis was inducibly expressed in Escherichia coli BL21 with a hexahistidyl N-terminus to permit its purification. Western blotting using the r6PGD-specific antiserum showed that the 6PGD protein is on the cell surface of H. parasuis. The characterization of 6PGD in H. parasuis pathogenesis involved as an adhesion and its immunogenicity in mice was further investigated. The adherence assay with H. parasuis and swine alveolar epithelial cells (SJPLC) pre-incubated with (His)66PGD and non-incubated SJPLC showed a noticeable reduction in the adhesion of H. parasuis in the (His)66PGD pre-incubated SJPLC compared to the non-incubated SJPLC. Further, the r6PGD protein induces the production of IL-8 and IL-6 by SJPLC. Furthermore, immunization with the r6PGD protein can provide the protective efficacy by 75% following intraperitoneal administration of a 5 × LD50 dose of H. parasuis SH0165, and elicited a good protective immune response, which demonstrated the importance of 6PGD to bacterial pathogenesis. Identification and characterization of the role of H. parasuis 6PGD in adhesion and immunogenicity will allow us to use this protein to develop new antimicrobial therapies and/or vaccines.  相似文献   

2.
Porins, expressed by Gram-negative bacteria, have several biological effects on host tissues or cells. The outer membrane protein P2 (OmpP2), a member of the porin family, has been identified as a multifunctional protein involved in the pathogenicity of Haemophilus parasuis. In the present study, it was shown that OmpP2 (0.5–10 μg/mL) from H. parasuis Nagasaki strain up-regulated mRNA expression of interleukin (IL)-1α, IL-1β, IL-6 and IL-8 in porcine alveolar macrophages (PAM, 3D4/31) in vitro, in a dose-dependent manner. Moreover, OmpP2 porin induced a more prolonged cytokine response in PAM than that of the lipooligosaccharide (LOS) from this microorganism. The data demonstrate that H. parasuis OmpP2 can stimulate proinflammatory cytokine mRNA expression, suggesting that this particular porin might play an important role in the pathogenesis of disease caused by H. parasuis.  相似文献   

3.
A real-time polymerase chain reaction (PCR) assay of the outer membrane protein (OMP) P2 gene was developed and used to test 97 putative Haemophilus parasuis pure cultures and 175 clinical tissue samples. With standard culture isolation as the gold standard, the diagnostic sensitivity and specificity of the PCR assay were determined to be 83% and 80%, respectively.  相似文献   

4.
Rectal smears of calves, cows and young bulls, as well as cloacal smears of house sparrows (Passer domesticus), from farms at the villages of Šumice and Troskotovice, Czech Republic, were examined for E. coli resistant to 12 antimicrobials. The resistant isolates were tested for antimicrobial-resistance genes and integrons. Totals of 40% (n = 183), 3% (n = 95), 0% (n = 33), and 9% (n = 54) of Escherichia coli isolates from calves, cows, young bulls and house sparrows, respectively, were antimicrobial resistant. The following genes were identified in cattle E. coli isolates: tetA, tetB (isolates resistant to tetracycline), blaTEM (beta-lactams), strA, aadA (streptomycin), sul1, sul2 (sulphonamides), and cat, floR (chloramphenicol). Seven of 16 antimicrobial-resistant calf isolates from the Šumice farm possessed class 1 integrons with the aadA1 gene cassette integrated, 1 kb in size. On the Troskotovice farm, eight of 57 antimicrobial-resistant calf isolates possessed class 1 integrons. Integrons of 1.5 kb with the dhfr1- aadA1 gene cassette were found in four isolates, followed by a 1 kb integron with the aadA1 gene found in three isolates, and a 1.7 kb integron with the dhfr17-aadA5 gene cassette and the phenotype ASSuTSxtNaCipCCfG. The prevalence of resistant E. coli in calves compared to adult cattle was much higher and probably was influenced by oral antimicrobial usage in calves, feeding with milk and colostrum from treated cows, as well as mechanisms unrelated to antimicrobial drug selection. Although house sparrows lived together with the cattle and came into contact with cattle waste on the farm, they were not infected by resistant E. coli isolates with the same characteristics as those found in cattle.  相似文献   

5.
The objective of this study was to isolate and identify yeast strains from broilers excreta and to evaluate in vitro their potential for probiotic use in animal production.Methods and resultsNine yeast strains were isolated and presumptively pre-identified by biochemical assays. These isolates were grouped in six different molecular profiles using PCR-fingerprinting technique. Each profile was identified by sequencing of the D1/D2 domains of the large subunit of the 26S rRNA gene. These yeasts were identified as: Trichosporon sp. (LV-2), Wickerhamomyces anomalus (LV-6), Pichia kudriavzevii (LV-8), Kodamaea ohmeri (LV-9) and Trichosporon asahii (LV-10). A pre-screening of the strains for probiotic use was based on their ability to agglutinate pathogenic micro-organisms. These yeast strains were characterized for specific growth rate, duplication time, their cell surface hydrophobicity, medium acidification, resistance to low pH (2.0, 2.5 and 3.0) and concentrations of bile salts (0.3% and 0.6%). The isolate of W. anomalus (LV-6) had the highest agglutinating and adherence capacity, a growth rate of 2.07 × 108 cfu/mL in 24 h at 30 °C, decreasing the medium pH from 6.5 to 5.23, a 25% hydrophobicity and an elevated capacity to grow under stress conditions.ConclusionsW. anomalus strain LV-6 showed the best characteristics for use as a probiotic candidate.Significance and impact of the Study:The data from this study helped in choosing a probiotic candidate from yeast to use in broiler production.  相似文献   

6.
Escherichia coli-associated diarrhoea is an important disease adversely affecting the pig industry. This study was conducted to investigate the frequency of virulence factors expressed by E. coli strains isolated from suckling pigs with diarrhoea in China. A total of 381 E. coli strains, obtained from 290 faecal samples from pigs on 38 farms, were tested for fimbriae (K88, K99, 987P, F41, F18, F17), non-fimbrial adhesins (AIDA-I, paa, CS31A, eae, saa), enterotoxin (LT-I, LT-II, STa, STb, EAST1), Shiga toxin (Stx1, Stx2, Stx2e), pathogenicity islands (HPI, LEE), α-haemolysin (hlyA), afa8 gene cluster (afaD, afaE) and sepA genes by PCR. Out of the 381 isolates, 206 carried at least one virulence gene. Of the 206 virulence positive isolates, the virulence factor genes detected were EAST1 (n = 120), irp2 (n = 59), paa (n = 50), STb (n = 41), AIDA-I (n = 34), LT-I (n = 23), ler (n = 11), hlyA (n = 9), K88 (n = 8), eae (n = 8), STa (n = 7), sepA (n = 6), F18 (n = 5), afaD (n = 3), afaE (n = 3), K99 (n = 2) and Stx2e (n = 1), with most isolates carrying multiple virulence genes. These results demonstrate that relatively few isolates from the study population express K88, K99, LT-I or STa, but that EAST1 (58%), irp2 (29%), AIDA-I (16.5%), paa (24%) and STb (20%) are frequent virulence factors expressed by E. coli strains isolated from suckling pigs with diarrhoea in China.  相似文献   

7.
Brucellosis is a highly contagious zoonotic infection affecting livestock and human beings. The disease has been reported worldwide except in few countries where it has been eradicated. The prevalence of brucellosis among cattle from 11 farms having a history of abortions was studied. A total of 481 samples comprising of blood, milk, vaginal swabs, vaginal discharges, placental tissues and fetal tissues were collected from 296 animals. Clinical samples were processed for the isolation of Brucella. Serum samples (n = 296) were tested by Rose Bengal Plate Test (RBPT) and indirect ELISA. A total of 90 (30.40%) and 123 (41.55%) samples were positive by RBPT and indirect ELISA, respectively. Also 27.02% samples were positive by both the tests. Brucella isolates (n = 8) were recovered from clinical samples using Brucella selective media. All the isolates demonstrated PCR amplification for the bcsp31 and IS711 genes. Amplification of Brucella abortus specific primer was demonstrated by all the isolates in AMOS PCR indicating isolates to be of either B. abortus biotype 1, 2 or 4. Risk factors for transmission of brucellosis among cattle population were studied by field surveys. It was observed that lack of awareness about brucellosis (OR = 8.739, P = 0.138) and inadequate floor space (OR = 0.278, P = 0.128) were crucial risk factors for transmission of bovine brucellosis.  相似文献   

8.
  相似文献   

9.
The increasing prevalence of antimicrobial resistances is now a worldwide problem. Investigating the mechanisms by which pets harboring resistant strains may receive and/or transfer resistance determinants is essential to better understanding how owners and pets can interact safely. Here, we characterized the genetic determinants conferring resistance to β-lactams and quinolones in 38 multidrug-resistant Escherichia coli isolated from fecal samples of dogs, through PCR and sequencing. The most frequent genotype included the β-lactamase groups TEM (n = 5), and both TEM + CTX-M-1 (n = 5). Within the CTX-M group, we identified the genes CTX-M-32, CTX-M-1, CTX-M-15, CTX-M-55/79, CTX-M-14 and CTX-M-2/44. Thirty isolates resistant to ciprofloxacin presented two mutations in the gyrA gene and one or two mutations in the parC gene. A mutation in gyrA (reported here for the first time), due to a transversion and transition (TCG → GTG) originating a substitution of a serine by a valine in position 83 was also detected. The plasmid-encoded quinolone resistance gene, qnrs1, was detected in three isolates. Dogs can be a reservoir of genetic determinants conferring antimicrobial resistance and thus may play an important role in the spread of antimicrobial resistance to humans and other co-habitant animals.  相似文献   

10.
We investigated the influence of administration of flax-seed oil on interaction of Lactobacillus plantarum – Biocenol? LP96 and Escherichia coli O8:K88ab:H9 in the gut of germ-free piglets. When compared to animals supplemented with L. plantarum, the counts of lactobacilli in the jejunal and ileal mucosa and in the intestinal content were significantly higher in LMK group (p < 0.0001). Inter-groups comparison of the counts of E. coli K88 adhering to the jejunal and ileal mucosa revealed a significantly decrease in LMK animals (p < 0.001; p < 0.05).  相似文献   

11.
ObjectivesTwo-dimensional (2D) speckle tracking echocardiography (STE) is a new angle-independent ultrasound technique based on tracking of speckles within the myocardium on 2D grayscale images. The aims of this prospective study were as follows: (1) to assess the variability of left ventricular peak systolic radial strain (St) and strain rate (SR) in awake dogs using STE (Protocol 1); and (2) to quantify these variables in a healthy canine population and compare them with tissue Doppler imaging (TDI)-based St and SR values (Protocol 2).BackgroundSt and SR may be assessed using TDI, which is limited by angle dependency.Animals, materials and methodsThirty-six STE examinations were performed on 6 healthy dogs for Protocol 1 and 37 healthy dogs were recruited for Protocol 2. In both studies, STE measurements were obtained offline from the right parasternal short-axis view by the same trained observer using automatic frame-to-frame tracking of grayscale speckle patterns.ResultsAll within- and between-day coefficients of variation were <10% (Protocol 1). In Protocol 2, St (46.7 ± 12.2%) and SR (2.7 ± 0.6 s−1) measured by STE were correlated with heart rate (p < 0.01), but not with the ratio of early mitral inflow velocity to early mitral annular velocity. There was a good correlation between STE and TDI for both St and SR values (p < 0.001).ConclusionsSTE is a repeatable and reproducible non-Doppler method for assessing radial St and SR. The combination of these indices with conventional echo-Doppler variables could provide a new approach for accurately quantifying canine systolic function.  相似文献   

12.
The aim of this study was to assess the prevalence of heartworm in domestic dogs in Madera and Fresno Counties, California, dependent on habitat and other host and environmental factors. Dogs were screened for presence of heartworm antigen using the PetChek® ELISA on blood samples (N = 519) collected at seven sites during April-July 2009. Eighteen dogs were heartworm antigen positive. Pearson Chi-square analyses were conducted testing the presence of heartworm antigen against the following independent variables: elevation range, percentage of time spent outdoors during the day, percentage of time spent outdoors during the night, pet coat length, weight class, prevention status, and sex. Dogs that spent at least 50% of their time outdoors during the day were significantly more likely to have heartworm that those who spent less time outside (N = 519, df = 1, p = 0.031). Overall prevalence (3.47%) was lower than expected, with Madera County having 3.8% positive samples and Fresno County 3.5%; this prevalence is lower than in many previous studies. The effect of time spent outdoors on heartworm prevalence was similar to previous studies. The impact of elevation on infection, though not significant, requires further investigation, as does the prevalence and viability of larval stages in mosquitoes.  相似文献   

13.
Bovine besnoitiosis is an economically important disease in cattle caused by the protozoan parasite Besnoitia besnoiti, which occurs endemically in many countries of Africa and Asia and is spreading in Europe. Serological identification of subclinically infected cattle is important to avoid the introduction of infected animals into naive herds. Here we determine the sensitivity and specificity of the PrioCHECK® Besnoitia Ab, a serological test recently introduced into the European market. Analytical specificity was examined using sera from animals experimentally infected with parasites related to B. besnoiti (n = 27). Three animals experimentally infected with Neospora caninum or Toxoplasma gondii showed inconclusive reactions in the ELISA (percent positivity relative to the positive control [PP] 10% ≤ 20%) while all other sera reacted negative (PP < 10%). An estimate of the diagnostic specificity was obtained by analysing field sera from bovine herds without besnoitiosis but with abortion problems associated to N. caninum (n = 403). The analysis revealed a specificity of 94.3% or 96.8% depending on the applied cut-off (PP 10% or 20%, respectively). Sensitivity was assessed with sera from 110 animals of a herd in Germany where clinical bovine besnoitiosis was first diagnosed in September 2008. A positive serological reference standard was defined regarding sera from animals as reference positive, if these animals had tested positive in at least two of a panel of three other serological tests (two different B. besnoiti immunoblots and one immunofluorescence antibody test) on both of two sampling dates, November 2008 and April 2009. A diagnostic sensitivity of 91.8% or 75.5% was determined for sera collected in November 2008 and a sensitivity of 82.7% or 50% for sera collected in April 2009 (cut-off PP 10% or PP 20%, respectively). The marked drop in sensitivity from November 2008 to April 2009 was predominantly observed in reference-positive cattle without clinical signs. We conclude that PrioCHECK® Besnoitia Ab is a valuable diagnostic tool to detect clinically infected animals. Thus it may be used to support control measures, e.g., for the separation of infected animals from the remaining herd to avoid a further transmission of the infection within the herd.  相似文献   

14.
This study characterized carriage and clinical pneumococcal isolates for serotypes, penicillin susceptibility, virulence genes and restriction fragment length polymorphism (RFLP) pattern of penicillin binding protein (PBP) genes. DNA fingerprint of isolates was generated by BOX-PCR. Majority of serotypes were 23F followed by 19F, 19A and 6A. Twenty-four percent of isolates were penicillin non-susceptible (PNSP). All of the targeted virulence genes were detected in all isolates with the exception of pili; 20.6% (n = 22) for PI-1 and 14.0% (n = 15) for PI-2. Of the 13 isolates which carried both PI-1 and PI-2, 10 were of clinical origin. Digested pbp-DNA produced three PBP-RFLP profiles for pbp1a (A1 to A3), six profiles for pbp2b (B1 to B6) and seven for pbp2x (X1 to X7) mostly in PNSPs. Based on BOX-PCR analysis, the majority of isolates were genetically diverse with a small number of potentially related isolates carrying pili genes. No obvious genotypic association was observed pertaining to carriage and clinical origin of isolates.  相似文献   

15.
Despite the importance of vegetative reproduction in annual tiller replacement, little is known about the patterns and timing of tiller recruitment from the bud bank, especially regarding fire return intervals and seasons of fire. We examined aboveground plant density, temporal patterns of tiller production, and belowground bud bank dynamics for Bouteloua gracilis (Willd ex. Kunth) Lag. ex Griffiths), Pascopyrum smithii (Rydb.) A. Löve, and Hesperostipa comata (Trin. & Rupr.) Barkworth following summer, fall, and spring prescribed fires at 2-yr, 3-yr, and 6-yr fire return intervals, and their interactions. Fire treatments were initiated in 2006, and buds were assessed July 2011 through July 2013. Density and number of reproductive B. gracilis tillers increased in 2013 following drought during 2012, unlike H. comata, which decreased reproductive tiller production. Irrespective of fire treatments, B. gracilis produced the most buds (8 ? 10 buds ? tiller? 1) and H. comata produced the least (2 ? 3 buds ? tiller? 1), with P. smithii producing an intermediate amount (6 ? 8 buds ? tiller? 1). Immediate B. gracilis and P. smithii bud mortality did not occur for all season and fire return interval treatments. However, H. comata bud mortality increased immediately following summer and fall prescribed fires. Three-yr fire return intervals increased active buds throughout the 2013 winter and growing season for B. gracilis and P. smithii relative to control plots and 2- and 6-yr fire return intervals. Fire stimulated bud activity of B. gracilis and P. smithii relative to nonburned plots. The aboveground and belowground response of H. comata indicated meristem limitations following fire treatments, illustrating greater vulnerability to fire for that species than B. gracilis and P. smithii.  相似文献   

16.
To investigate effective new rabies vaccines, a fusion protein consisting of the rabies virus (RV) glycoprotein and the heat-labile enterotoxin B subunit of Escherichia coli (LTB) was successfully constructed and delivered in a live attenuated Salmonella strain LH430. Mice were immunised with LH430 carrying pVAX1-G, pVAX1-G-LTB or pVAX1-ori-G-LTB. The antibody titres of mice immunised with oral LH430 carrying pVAX1-G-LTB or pVAX1-ori-G-LTB were significantly higher than those of pVAX1-G-immunised mice. The results of the challenge with the rabies virus standard strain (CVS-11) showed that the LH430 strain carrying the G-LTB gene induced immunity and elevated IL-2 levels in immunised mice (7P < 0.01), whereas LH430 carrying pVAX1-G did not contribute to protection. These results show that LH430 carrying recombinant G-LTB could provide overall immunity against challenge with CVS-11 and should be considered to be a potential rabies vaccine.  相似文献   

17.
Mannan-binding lectin (MBL), a pattern recognizing serum protein, participates in the innate immune system of mammals as an opsonin. In humans, single-nucleotide polymorphisms (SNPs) in MBL2 gene were found to cause various innate immune dysfunctions. In the present study, we discovered three single-nucleotide polymorphisms of the MBL1 gene in Chinese native cattle and analyzed their associations with milk traits. By screening the genetic variation of MBL1 in 1053 individuals of three Chinese native cattle breeds including China Holstein, Luxi Yellow and Bohai Black using created restriction site–polymerase chain reaction (CRS–PCR), PCR–restriction fragment length polymorphism (PCR–RFLP) and DNA sequencing techniques, three new SNPs, g.855G>A, g.2651G>A and g.2686T>C, were found to have allele frequencies of 0–12.65%, 24.07–42.39% and 56.95–73.68%, respectively. While SNP g.855G>A is located within intron ?, the other two SNPs reside in the exon II region with one mutation being non-synonymous (GTT (Val) > ATT (Ile)) and the other synonymous (GCT (Ala) > GCC (Ala)). Among the 596 Chinese Holstein cattle with at least 3 lactation Dairy Herd Improvement (DHI) records, eight different haplotypes and 19 genotype combinations were detected. Statistical analyses revealed no correlation between either g.855G>A or g.2686T>C and somatic cell score (SCS), however significant association was found between g.2651G>A and SCS, suggesting a possible role of this SNP in the host response against mastitis. Our data also suggested that the combined genotypes of GGC/AAC with the lowest SCS, AAT/AAT with the highest protein content and AGC/AGC with the highest 305-d milk yield were favorable combinations for mastitis resistance and milk production traits. Therefore, GGC/AAC, AAT/AAT and AGC/AGC can be used as possible candidates for marker-assisted selection in the dairy cattle breeding program.  相似文献   

18.
This study was conducted to explore the epidemiological and molecular differences between carbapenem-susceptible Acinetobacter baumannii (CSAB) and carbapenem-resistant A. baumannii (CRAB) isolates. Thirty-two CSAB and 55 CRAB isolates were collected in 2010. By multilocus sequence typing analysis, 31 (56%) CRAB isolates and 11 (34%) CSAB isolates belonged to ST2. Twenty-one (38%) CRAB isolates, and 4 (13%) CSAB isolates belonged to a new type, ST129. The blaIMP, blaVIM, and blaOXA-58-like were not detected in our study isolates. blaOXA-23 and blaOXA-24/40-like were not detected in all CSAB isolates. On the contrary, blaOXA-23 was detected in 51 (93%) CRAB isolates. Class 1 integron was detected in 19 (35%) CRAB isolates and 8 (25%) CSAB isolates (p > 0.05). In conclusion, the ST2 and ST129 were the major sequence types in both CSAB and CRAB isolates. The blaOXA-23 is the primary carbapenem-resistance gene in CRAB isolates from hospitalized patients and the specimens collected from hospital environment.  相似文献   

19.
Biofilms are surface-associated microbial communities, which are encased in self-synthesized extracellular environment. Biofilm formation may trigger drug resistance and inflammation, resulting in persistent infections. Haemophilus parasuis is the etiological agent of a systemic disease, Glässer's disease, characterized by fibrinous polyserositis, arthritis and meningitis in pigs. The purpose of this study was to examine the correlation between biofilm and antibiotic resistance among the clinical isolates of H. parasuis. In the present study, we tested biofilm-forming ability of 110 H. parasuis isolates from various farms using polystyrene microtiter plate assays. Seventy-three isolates of H. parasuis (66.4%) showed biofilm formation and most of them performed weak biofilm-forming ability (38/73). All isolates were tested for antimicrobial susceptibility to 18 antimicrobial agents by the broth microdilution method. H. parasuis isolates showed very high resistance (>90%) to sulfanilamide, nalidixic acid, and trimethoprim. Resistance to eight antibiotics such as penicillin (41.1% vs 8.1%), ampicillin (31.5% vs 8.1%), amoxicillin (28.8% vs 5.4%), gentamicin (46.6% vs 24.3%), cefazolin (19.2% vs 2.7%), doxycycline (19.2% vs 8.1%), cefotaxime (11% vs 2.7%), and cefaclor (13.7% vs 5.4%) was comparatively higher among biofilm producers than non-biofilm producers. Pulsed-field gel electrophoresis (PFGE) analyses could distinguish various isolates. Our data indicated that H. parasuis field isolates were able to form biofilms in vitro. In addition, biofilm positive strains had positive correlation with resistance to β-lactams antibiotics. Thus, biofilm formation may play important roles during H. parasuis infections.  相似文献   

20.
  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号