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1.
Pasteurella multocida serogroup B:2, a causative agent of haemorrhagic secpticaemia (HS) in cattle and buffalo especially in tropical regions of Asia and African countries, is known to possess a type IV fimbriae (pili) as one of the virulent factors. In the present study, ptfA gene encoding for type IV fimbrial subunit of P. multocida serogroup B:2 (strain p52), an Indian HS vaccine strain, has been cloned and over-expressed in recombinant Escherichia coli. The recombinant type IV fimbrial subunit protein (～31kDa) including N-terminus histidine tag was purified under denaturing condition and confirmed by western blotting. A homology model of HS causing P. multocida serogroup B:2 fimbrial subunit has also been discussed. The study indicated the potential possibilities to use the recombinant fimbrial protein in developing HS subunit vaccine along with suitable adjuvant.  相似文献   

2.
The susceptibility of Pasteurella multocida to killing by serum and the ability of protective vaccines to stimulate this mechanism of immunity in mice were investigated. P. multocida type of bovine origin was used to prepare a vaccine incorporating heat killed organisms and for homologous infection of mice. Bactericidal capacity and ELISA antibody titres were determined for individual mouse sera. Protection was clearly associated with bactericidal antibodies raised by vaccination. The bactericidal assay may be useful as a rapid, simple screening test of vaccinated mice for functional protective antibody levels.  相似文献   

3.
Identification and estimation of the prevalence of Pasteurella multocida organisms in different animal and avian species in India during November 2000 to July 2003 was carried out. Out of 418 samples collected from different outbreaks suspected to be caused by P. multocida, a total of 206 bacterial cultures were identified as P. multocida on the basis of cultural, morphological and biochemical characteristics. All the 206 cultures were isolated from different domestic animal species (cattle, buffalo, sheep, goat, pig and rabbit), avian species (chicken, duck, quail, turkey, goose) and wild animals such as leopard and deer. Serotyping of P. multocida cultures revealed the presence of various serotypes (A:1, A:3, A:1,3, A:4, B:2, D:1 and -:1) among the livestock population. P. multocida polymerase chain reaction (PCR) assay applied on different forms of bacterial cultures (bacterial culture lysate, direct bacterial colony and mixed bacterial culture lysate) yielded an amplified product of approximately 460 bp specific for P. multocida. The results of PCR assay correlated well with conventional methods of identification. The present investigation revealed the presence of varied serotypes among livestock and PCR assay was found to be useful for rapid, sensitive and specific diagnosis of pasteurellosis in animals and avian species.  相似文献   

4.
AIMS: To examine pigs at slaughter in New Zealand for the presence of Pasteurella multocida, and to determine for isolates, their biochemical profiles, somatic and capsular types, and the presence or absence of the HSB and toxA genes, associated with haemorrhagic septicaemia (HS) and progressive atrophic rhinitis (PAR), respectively. METHODS: Swabs from 173 lungs, 158 palatine tonsils and 82 nasal passages of pigs at two abattoirs in New Zealand were cultured for P. multocida using conventional techniques, and isolated colonies were subjected to biochemical tests for identification of biovars. Somatic serotyping was conducted using an agar gel immunodiffusion (AGID) test. Polymerase chain reaction (PCR) assays were used to confirm phenotypic identification of colonies using species-specific primers, capsule type using serogroup-specific primers and multiplex PCR, and to test for the presence of HSB and toxA genes. RESULTS: Pasteurella multocida was isolated from 11/173 (6.4%) lung, 32/158 (20.2%) palatine tonsil and 5/82 (6.1 %) nasal swab samples, a total of 48 isolates from 413 samples (11.6%). Isolation rates per farm ranged from 1-53% of tissue samples collected from pigs 5-6 months of age. On phenotypic characterisation, isolates were allocated to seven main biovars, viz 1, 2, 3, 5, 9, 12, and a dulcitol-negative variant of Biovar 8, the majority (30/48) being Biovar 3. Of the 42 isolates for which somatic serotyping was conducted, 10% were Serovar 1, 79% were Serovar 3, 2% were Serovar 6,1, 2% were Serovar 12, and 7% could not be typed. All 48 isolates were confirmed as P. multocida using a species-specific PCR. In the capsular multiplex PCR, 92% of isolates were Capsular (Cap) type A, 2% were Cap D, and 6% could not be typed. None of the samples were positive for the HSB or toxA genes. CONCLUSION: Serovars or capsular types of P. multocida associated with HS or PAR in pigs were not detected. Establishment of species-specific, capsular and toxin PCR assays allowed the rapid screening of isolates of P. multocida, while serotyping provided an additional tool for epidemiological and tracing purposes.  相似文献   

5.
Repetitive extragenic palindromic (REP)-PCR (polymerase chain reaction), enterobacterial repetitive intergenic consensus (ERIC)-PCR, and single primer PCR assays were employed to characterize 66 strains of Pasteurella multocida serogroup A:1 isolated from avian species belonging to different regions of India. REP-PCR resulted in amplification of REP sequences from the genome which were in the range of approximately 200 to approximately 3000 bp and accounted for a total of 54 distinguishing profiles (D=0.99). ERIC-PCR analysis also generated amplified products in the range of approximately 200 to approximately 3200 bp categorizing strains into a total of 50 different profiles (D=0.98). Amplification of repetitive regions using a microsatellite primer (GTG)(5), resulted in clear distinctive bands ranging from approximately 200 to approximately 2400 bp. Strains were assigned to 43 profiles (D=0.96). No correlation could be drawn between genotypic profiles and avian hosts with their geographical area of origin. Avian strains of P. multocida serogroup A:1 were found to be highly heterogeneous with diverse profiles. REP-PCR was found to be highly discriminatory and simple method for differentiation of phenotypically similar strains. The present study also indicated that PCR based amplification of repetitive regions of P. multocida is a rapid technique with good discrimination and could be employed directly for routine typing of field isolates from fowl cholera outbreaks.  相似文献   

6.
The efficacy of two different types of commercial vaccines against PRRSV (Euro-type) was evaluated based on clinical parameters upon challenge as well as post-challenge virological profiles (viremia and viral load in tissues upon necropsy, measured in both cases by quantitative real time PCR). In an attempt to establish correlates of protective immunity, two commonly proposed parameters predictive of immunity were measured: (1) serologic responses (ELISA and neutralizing antibodies), (2) frequency of gamma interferon-producing cells in peripheral blood mononuclear cell fraction. The vaccines compared consisted of two commercially available products that are regularly marketed in Spain: one modified live virus and one killed vaccine. The efficacy assay was carried out by vaccinating twice 3 weeks apart groups of 5 and-a-half month-old female swine and then challenging them with a European type 1 PRRSV strain (Lelystad). The results obtained indicate that the modified live virus vaccine was the only type of vaccine capable of establishing protective immunity, as measured by viral load in blood and tissues. The killed vaccine, in spite of this product evoking a spontaneous interferon-gamma response and post-challenge titers of virus-neutralizing antibody, evoked no measurable protective immunity. In the case of the modified live vaccine, the protection exhibited did not appear to be based on humoral but rather on cell-mediated immunity.  相似文献   

7.
The applicability of conventional and molecular methods for rapid detection and differentiation of Pasteurella multocida serogroup B isolates involved in an outbreak of haemorrhagic septicaemia affecting Indian buffaloes, was studied. Five isolates were obtained and were subjected to phenotypic and genotypic characterization. None of the five isolates could be differentiated on the basis of cultural, biochemical, pathogenicity and antimicrobial sensitivity patterns. Polymerase chain reaction (PCR)-based techniques were found to be specific and sensitive for rapid detection and differentiation of isolates. Repetitive extragenic palindromic (REP-) PCR, enterobacterial repetitive intergenic consensus (ERIC-) PCR and single-primer PCR differentiated all the five isolates into different profiles. All the isolates involved in the outbreak were found to have a genetic profile different from standard P. multocida strain (P52). However, three isolates had similar profiles, whereas each of the remaining two had a different profile. The study indicates the involvement of multiple strains of P. multocida in a single outbreak of haemorrhagic septicaemia in buffaloes. The results also indicate that molecular methods of detection and typing are superior to conventional methods for rapid epidemiological investigations of haemorrhagic septicaemia.  相似文献   

8.
Pasteurella multocida and bovine respiratory disease   总被引：1，自引：0，他引：1  
Pasteurella multocida is a pathogenic Gram-negative bacterium that has been classified into three subspecies, five capsular serogroups and 16 serotypes. P. multocida serogroup A isolates are bovine nasopharyngeal commensals, bovine pathogens and common isolates from bovine respiratory disease (BRD), both enzootic calf pneumonia of young dairy calves and shipping fever of weaned, stressed beef cattle. P. multocida A:3 is the most common serotype isolated from BRD, and these isolates have limited heterogeneity based on outer membrane protein (OMP) profiles and ribotyping. Development of P. multocida-induced pneumonia is associated with environmental and stress factors such as shipping, co-mingling, and overcrowding as well as concurrent or predisposing viral or bacterial infections. Lung lesions consist of an acute to subacute bronchopneumonia that may or may not have an associated pleuritis. Numerous virulence or potential virulence factors have been described for bovine respiratory isolates including adherence and colonization factors, iron-regulated and acquisition proteins, extracellular enzymes such as neuraminidase, lipopolysaccharide, polysaccharide capsule and a variety of OMPs. Immunity of cattle against respiratory pasteurellosis is poorly understood; however, high serum antibodies to OMPs appear to be important for enhancing resistance to the bacterium. Currently available P. multocida vaccines for use in cattle are predominately traditional bacterins and a live streptomycin-dependent mutant. The field efficacy of these vaccines is not well documented in the literature.  相似文献   

9.
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10.
Gnotobiotic pig antisera to purified toxoid from a capsule type A or D strain of Pasteurella multocida contained large quantities of antitoxin but comparatively little antibody to a crude lysate of P. multocida. These sera given intraperitoneally to further pigs were almost completely protective against turbinate atrophy after intranasal inoculation of dilute acetic acid and infection with type D toxigenic P. multocida. In contrast, antisera to a crude lysate or bacterin of toxigenic P. multocida which contained large titres of antibody to P. multocida lysate, but no detectable antitoxin, were not protective. Colonisation by toxigenic P. multocida was significantly reduced in protected pigs and was similar to colonisation by nontoxigenic P. multocida in pigs untreated or treated with dilute acetic acid. These results indicated (1) that antitoxin was protective and cross protective between toxins from different capsule types; and (2) that the toxin was the main colonisation factor produced by toxigenic bacteria in the acetic acid model of infection and that immunity to it did not eliminate infection.  相似文献   

11.
多杀性巴氏杆菌是一种常见病原菌，其血清型众多（16种血清型），在毒力及流行病学等方面的生物学特性差异很大。分子生物学鉴定和特性分析方法能直接揭示其基因型上的差异，具有高灵敏度和可靠性。本文综述了多杀性巴氏杆菌的PCR、菌落杂交和16S rRNA基因测序等鉴定方法以及限制性酶切分析、核糖体分型、脉冲场凝胶电泳、PCR指纹等特性分析方法。  相似文献   

12.
Turkey poults were vaccinated with combinations of two different germling preparations and three adjuvants (N-acetylmuranyl-L-alanyl-D-isoglutamine, Pasteurella multocida lipopolysaccharide [LPS], and avridine) at 1 and 2 weeks of age, and their immunity was challenged by sublethal exposure to aerosols of Aspergillus fumigatus conidia at 1 month of age. Fewer turkeys in the groups given vaccines prepared from germlings grown on Dorset's and Henley's medium (D&H) had organisms in lung tissue at 2 weeks after challenge exposure as compared with those vaccinated with germling grown on neopeptone dialysate (Neo). The LPS of P. multocida appeared to be the most efficacious of the adjuvants in the D&H vaccine group, as A. fumigatus was isolated from only one of eight turkeys in this group; the number of organisms per gram of lung tissue was low compared with other vaccine groups at 2 weeks after challenge exposure; and poults given D&H vaccine with LPS as adjuvant had less-severe lung lesions than other groups. These differences in lung lesions were more marked at 2 weeks than at 8 weeks after challenge exposure. The only difference among other parameters in the vaccinated turkeys was lower heterophil counts in the turkeys given D&H-prepared vaccines than in unvaccinated controls. This was probably due to less-severe infections resulting from protective effects of these vaccines.  相似文献   

13.
Haemorrhagic septicaemia (HS) is caused by specific serotypes of Pasteurella multocida and is one of the major economic diseases of cattle and buffalo in South East Asia. Definitive diagnosis of the disease-causing organism with the available methods is labour intensive and not totally reliable, consequently, an ELISA system to identify P multocida organisms which cause HS was developed. One hundred and twenty-four P multocida isolates were tested, 58 were type strains and 66 were field isolates. Analysis of these strains indicated the assay had a specificity of 99 per cent and sensitivity of at least 86 per cent. The sensitivity could be an underestimate, as five isolates assumed to be false negative reactions may not all be HS-causing strains. The HS ELISA provides a rapid, simple, accurate and inexpensive diagnostic assay for identification of HS causing organisms but does not represent a new typing system for P multocida. This assay will also enable countries to assess the impact of HS more accurately.  相似文献   

14.
Fowl cholera continues to be of concern to the poultry industry, especially for turkey growers. This disease costs the turkey industry millions of dollars annually. In order to develop improved live attenuated vaccines or subunit vaccines, the outer-membrane proteins of Pasteurella multocida were examined with the use of proteomics. Of the 11 proteins total present in an outer-membrane subfraction of P. multocida, four additional proteins were identified, completing the composition of the detergent-soluble cross-protective protein fraction. These additional four proteins include protective bacterial surface antigen, OMA87 (Accession no. 15603857); heme-hemopexin receptor, HemR (Accession no. 15602441); lactate permease, LctP (Accession no. 15603717); and heptosyl transferase F, RfaF (Accession no. 15603709). Both the Oma87 and the HemR proteins would be of interest for subunit and modified live vaccine studies, respectively, because of their purported roles as virulence factors for P. multocida.  相似文献   

15.
Turkeys developed a high level of protective immunity and serum anti-Pasteurella multocida antibody when vaccinated with the Clemson University (CU) strain of P. multocida via the following routes: wing-web puncture with the recommended dosage or 1:10 dilution of this dosage, subcutaneous, crop injection, cloacal perfusion of the bursa of Fabricius, and intratracheal. The development of immunity after vaccination via the wing-web puncture and subcutaneous routes was dose-dependent: the lower dosages induced less serum antibody and protective immunity. Immunity was greatest 3 weeks after vaccination via wing-web puncture, although immunity was evident as early as 4 days after vaccination. A high level of antibody and protective immunity with no adverse reactions was induced by vaccinating twice, either by the drinking water followed by a similar dosage parenterally or by a parenteral route followed by the drinking water; these regimens were similar to one vaccination in the drinking water. Vaccination via wing-web puncture and possibly subcutaneous injection and bursal perfusion could be used for flocks with a history of a high mortality after vaccination with the CU vaccine in the drinking water and suspected of being immunosuppressed.  相似文献   

16.
Pasteurella multocida is responsible for major animal diseases of economic significance in both developed and developing countries whereas human infections related to this bacterium are infrequent. Significantly, development of a carrier status or latent infections plays a critical role in the epidemiology of these diseases. Aiming at increased knowledge of these infections, we examine potential diagnostic and selected typing systems for investigating diseases caused by P. multocida. Detection of P. multocida from clinical specimen by; (i) isolation and identification, (ii) polymerase chain reaction (PCR), iii) specific hybridisation probes, (iv) serological tests and (v) other alternative methods is critically evaluated. These detection systems provide a wide spectrum of options for rapid diagnosis and for detecting and understanding of latent infections in herd/flock health control programmes, though PCR methods for detecting P. multocida in clinical specimen appear increasingly preferred. For establishing the clonality of outbreak strains, we select to discuss macromolecular profiling, serotyping, biotyping, restriction enzyme analysis, ribotyping and multiplex PCR typing. Although P. multocida infections can be rapidly diagnosed with molecular and serological tests, isolation and accurate species identification are central to epidemiological tracing of outbreak strains. Our review brings together comprehensive and essential information that may be adapted for confirming diagnosis and determining the molecular epidemiology of diseases associated with P. multocida.  相似文献   

17.
Haemorrhagic septicaemia (HS) is an acute disease of cattle and buffaloes caused by Pasteurella multocida 6:B. Outbreaks of the disease have been closely associated with carrier animals that transmit the organism to susceptible animals during stressful condition. This study was conducted to determine whether goats exposed intranasally to P. multocida 6:B can transmit the organism to contact goats. Thirty-six healthy local Katjang goats were divided into four groups and goats of groups 1 and 3 were each inoculated intranasally with a 1-ml inoculum that contained 1 x 10(9) CFU/ml of live P. multocida 6:B. Following the exposure, all goats of groups 3 and 4 were injected with dexamethasone at the rate of 1 mg/kg for three consecutive days. At the end of the dexamethasone treatment, goats of groups 1 and 2 were commingled but kept separate from goats of groups 3 and 4, which were commingled in another pen. Three surviving goats from each group were killed on days 7, 14 and 21 post-exposure for postmortem examination. Naso-pharyngeal mucus and heart blood were collected on swabs. Tissues from lungs, lymph nodes and tonsils were collected for bacteriological isolation and identification. Only one goat of group 3 died 6 days post-exposure showing clinical signs and lesions typical of HS. Other goats showed mild signs of upper respiratory tract infection. Goats of all groups developed acute mild pneumonic lesions, however, those treated with dexamethasone had significantly (P < 0.05) more extensive lesion scoring based on the lesion scoring system. P. multocida 6:B was isolated from the nasal mucosa and lung lesions of exposed and contact goats not treated with dexamethasone. Exposed and contact goats treated with dexamethasone carried the organism for 21 days. P. multocida isolation from heart blood was made only from exposed and contact goats treated with dexamethasone. P. multocida was isolated from the lymph node of the goat that died during the experiment.  相似文献   

18.
Immunity assays on sheep sera using passive mouse protection tests showed that vaccines containing more than 4 strains of Pasteurella multocida did not give a good immunity. The immune response was not enhanced by the use of an oil adjuvant, and high concentrations of bacteria had only a partial positive effect. Attempts to extract selectively the protection-inducing antigen(s) from P. multocida by veronal, phenol or potassium thiocyanate extraction were unsuccessful. Furthermore, it was found that sheep antisera to the recognized type strains of P. multocida afforded only limited protection against a number of field strains. We concluded from this that successful immunization against ovine pasteurellosis will depend on either the identification of a strain of P. multocida that gives a wide spectrum of immunity or the discovery of a live mutant suitable for vaccine production and the definition of cultural conditions that promote the expression of a common immunizing antigen.  相似文献   

19.
Killed vaccines in oil emulsions are critical components of breeder and layer vaccination programs. Current vaccination sites are limited, and each has inherent problems. Oil emulsion vaccines are associated with increased condemnations of spent fowl when vaccines are injected intramuscularly into the breast. In an attempt to reduce tissue reaction when injected into the breast muscle, a commercially available Pasteurella multocida bacterin was heated to 41 C (100 F) for 5 hr prior to administration. A second treatment group was injected with the same bacterin at room temperature, 25 C. The vaccine was injected into the breast muscle at 10 and 18 wk of age into white Leghorn hens. Seroconversion was evaluated using P. multocida enzyme-linked immunosorbent assay (ELISA) at 10, 18, and 24 wk. Treatment and control groups were euthanized and lesions scored at 24 wk of age. One replicate was challenged with type 1 P. multocida at 24 wk of age. Lesion scores for the heated vaccine group were significantly lower than the room temperature vaccine. ELISA titers were not significantly different at 24 wk between the two treatment group; however, a significant rise in antibody titer was observed at 18 wk in the group that was injected with the heated vaccine. Survivability to challenge was improved in birds injected with the heated vaccine. Results suggest that heating of a P. multocida bacterin reduces local tissue reaction without having a deleterious effect on immunity as measured by ELISA and challenge.  相似文献   

20.
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