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1.
Avian pathogenic strains of Escherichia coli cause a number of extraintestinal diseases in poultry, including airsacculitis and colisepticemia. Expression of O78 lipopolysaccharide (LPS) is frequently associated with pathogenic isolates. Salmonella, a common poultry contaminant, is a major public health concern. The purpose of this work was to develop an E. coli vaccine for poultry with the use of an attenuated Salmonella typhimurium carrier that would benefit both the bird and the consumer. Orally administered attenuated S. typhimurium delta cya delta crp strains have been shown to provide excellent protection against wild-type Salmonella challenge in chickens. This work describes the construction of a delta cya delta crp derivative of an avian pathogenic S. typhimurium that expresses both the homologous group B determinants (O1,4,5,12) and the heterologous E. coli O78 LPS O antigens. This was accomplished by inserting the E. coli rfb region, which encodes the genes required for O78 expression, into the chromosomal cya gene of S. typhimurium, creating a defined deletion/insertion mutation. A delta crp mutation was introduced in a subsequent step. Expression of both O antigens was stable in vitro and in vivo. Vaccination of white leghorn chicks at day of hatch and 14 days with the recombinant vaccine strain induced serum immune responses against both S. typhimurium and E. coli LPS and protected the birds against subsequent challenge with an avian pathogenic E. coli O78 strain. Introduction of a mutation in rfc, which encodes the O antigen polymerase, reduced the chain length of the S. typhimurium LPS without affecting the expression of O78. The rfc mutation further enhanced the ability of the vaccine strain to protect chickens against E. coli challenge.  相似文献   

2.
In the day-old chick, physiological stress in the form of high or low temperatures or feed and water deprivation either interfered with colonization or altered the protection provided by normal intestinal microflora against subsequent challenge with Salmonella typhimurium. The response was more obvious after individual exposure of chicks to salmonella than after exposure to infected seeders. Physiological and dietary stress at 2 weeks of age did not appreciably alter the S. typhimurium excretion pattern in treated chicks. The protective microflora appears to be quite stable under these stress conditions.  相似文献   

3.
Avian pathogenic Escherichia coli strains are associated with a variety of extraintestinal poultry diseases, including airsacculitis, colisepticemia, and cellulitis. A number of E. coli serotypes are associated with these diseases, although the most prevalent serotype is O78. Fimbrial proteins expressed by these strains appear to be important virulence factors, including type 1 fimbriae, P fimbriae, and curli. We have been working to develop an effective vaccine to protect chickens against these diseases. We have previously shown that an attenuated Salmonella typhimurium strain expressing O78 lipopolysaccharide provides protection against challenge with an O78 avian pathogenic E. coli strain. In this work, we have constructed an attenuated S. typhimurium that expresses both the O78 lipopolysaccharide and E. coli-derived type 1 fimbriae. In these studies, chickens were vaccinated at day of hatch and again at 2 wk of age. Birds were challenged at 4 wk of age. We found that the vaccine candidate provided significant protection against airsacculitis as compared to untreated controls or birds vaccinated with an attenuated S. typhimurium that did not express any E. coli antigens. In a separate experiment, challenged vaccinates showed significant weight gain compared to challenged nonvaccinates. We were not able to demonstrate protection against E. coli O1 or O2 serotype challenge, nor against challenge with wild-type S. typhimurium.  相似文献   

4.
To test the interaction between various species of bacteria and Salmonella serovar typhimurium (S. typhimurium), the population of S. typhimurium was measured in the cecum of gnotobiotic chickens in the presence of Escherichia coli (E. coli) and one of the four intestinal bacteria; Lactobacillus acidophilus. Clostridium perfringens, Bifidobacterium thermophilum and Bacteroides vulgatus. Competitive exclusion of S. typhimurium by di-flora chicken was not demonstrated. But the population of S. typhimurium was temporarily suppressed in di-flora chickens with E. coli and L. acidophilus. In penta-flora chickens with E. coli and these four intestinal bacteria, the population of S. typhimurium was suppressed for only 2 days. In normalized chickens, the population of S. typhimurium was markedly suppressed.  相似文献   

5.
A native intestinal microflora of chickens which is protective against salmonella readily transferred to penmates and apparently to birds in adjacent pens. The microflora not only minimized infection resulting from exposure following colonization of the gut with microflora, but significantly abbreviated the period of infection when introduced after a salmonella infection was established in chicks. A microflora with undiminished protective activity, sensitive to only a few commonly used antibacterials, was established in a SPF-Cofal/Marek-negative population. Intestinal microflora from mourning doves was at least partially effective in protecting chicks against a naladixic-acid-resistant strain of Salmonella infantis. In limited tests with 2 of 3 sources of protective microflora, the growth rate of chicks in the absence of salmonellae was significantly improved. A hypothesis involving specificity of attachment between the glycocalyces of the protective microflora and of the intestinal mucosa is offered as the likely mechanism of protection.  相似文献   

6.
Chicks were treated orally on the day of hatch with either fresh or frozen competitive exclusion (CE) cultures (native gut microflora). Chicks were fed either unmedicated feed or one of five commercial broiler starter rations or nine experimental feed mixtures containing varying amounts and combinations of anticoccidial and antimicrobial medicaments. After 2 days, they were challenged with approximately 10(6) colony-forming units of a nalidixic-acid-resistant strain of Salmonella typhimurium. Six days later, chicks were sacrificed and ceca were analyzed for S. typhimurium. Colonization of 2-day-old chicks was prevented or at least greatly reduced in most instances by treatment of chicks with a CE culture, but the efficacy of CE broth cultures stored at -70 C diminished over time. Not all CE cultures tested gave equal protection against Salmonella colonization, and CE cultures were more susceptible to some feed additives than others. Of the commercial or experimental feed tested, only the feed containing the combination of nicarbazin and bacitracin interfered with the protective effect of the CE culture.  相似文献   

7.
Chickens that have considerable resistance to Salmonella typhimurium or Escherichia coli infection by early development of a native intestinal microflora shed these bacteria following aerosol exposure to Mycoplasma gallisepticum and/or infectious bronchitis virus. Administration of cyclophosphamide to similarly treated chickens induced slight shedding of these bacteria, and the combination of cyclophosphamide and respiratory agents magnified the shedding rate. These agents also influenced the isolation rate of E. coli and S. typhimurium from the trachea and air sacs.  相似文献   

8.
Resistance of young chicks to Campylobacter fetus subsp. jejuni was substantially increased by early exposure to native gut microflora. Protection was demonstrated against two human isolates and a chicken isolate of C. fetus subsp. jejuni. Significant protection against the chicken isolate was observed throughout a 91-day test period. Infection reached 100% (25/25) in the untreated group at 56 days of age and only 4% (1/25) in the group treated with native gut microflora. Campylobacter fetus subsp. jejuni was isolated from the ceca and less frequently from the gall bladder and liver of chicks that actively shed the bacteria. Cultures of feces from chicks reared on wood-shavings litter were often negative, suggesting that culturing litter as an indicator of infection has limited value.  相似文献   

9.
Isolation and characterization of avian rotaviruses   总被引:5,自引:0,他引:5  
Rotaviruses were isolated from intestinal contents obtained from flocks of turkey poults and pheasant poults with diarrhea and from different age groups of chickens showing various signs of intestinal disorders. The incorporation of 5 micrograms trypsin/ml in the inoculum and medium was essential for virus isolation in chicken kidney cells. All isolates were identified as rotaviruses by fluorescent-antibody technique using a National Institutes of Health reference rotavirus antiserum against human rotavirus strain "D," Type 2. Negative-staining and transmission electron microscopy showed the presence of rotavirus particles. Furthermore, polyacrylamide gel electrophoresis pattern of viral RNA segments of our isolates confirmed that they are rotaviruses. Seven-day-old turkey poults could be infected with turkey and chicken rotavirus isolates; in contrast, chicks of the same age were refractory.  相似文献   

10.
对从天津地区分离到的71株鸡大肠杆菌的部分生物学特性包括致病性、血清型、耐药特性和免疫原性等进行了研究。结果表明其中60株为致病性菌株,占分离菌株的84.5%;60个致病性菌株共定型出45个菌株,分属O1、O2、O5、O6、O20、O45、O53、O74、O75、O78、O88、O89、O92、O107、O111、O145等16个血清型,其中O78、O88、O2、O45、O53和O145为优势血清型,占定型菌株的73.4%;试验菌株具有广泛的耐药性,60个致病性菌株均为多重耐药。免疫原性测定试验结果表明,O2、O78、O88血清型菌株均可对相同血清型菌株提供很好的保护,但3个血清型菌株之间缺乏有效的保护。  相似文献   

11.
This investigation was designed to study the effect of exposure time, pH, age of bird, and native intestinal microflora on the in vitro attachment of Salmonella typhimurium to the ceca of chickens. Ceca were surgically removed from chickens immediately after euthanasia, and the interiors were exposed to S. typhimurium as intact ceca with contents, intact ceca rinsed free of contents, or inverted rinsed ceca. Attachment of S. typhimurium was slightly higher in washed than in unwashed ceca. Neither pH nor age of chicks affected attachment of the organism to ceca. There was no difference in attachment of salmonellae to inverted washed ceca after 1, 2, 3, 5, and 10 min exposure, but a one log difference was noted between 10 min and 30 sec.  相似文献   

12.
A commercial infectious bronchitis virus (IBV) vaccine of the Massachusetts 41 strain was injected in embryonating chicken eggs on embryonation day (ED) 18. The IBV vaccine was pathogenic for embryos, and it was passaged in chicken kidney tissue culture to reduce the pathogenicity. At the 40th tissue culture passage (P40-IBV), the virus became apathogenic for the embryos. Maternal antibody-positive or -negative chicks hatching from eggs injected with P40-IBV developed antibody to IBV and were protected against challenge exposure at 4 weeks of age with virulent Massachusetts 41 IBV. Although P40-IBV protected chicks when administered on ED 18, this virus did not protect chicks well if given at hatch. When combined with the turkey herpesvirus (HVT), P40-IBV given on ED 18 did not interfere with the protection against challenge exposure with virulent Marek's disease virus, nor did the presence of HVT interfere with protection by P40-IBV. Thus, under laboratory conditions, IBV vaccine could be combined with HVT to form a bivalent embryonal vaccine.  相似文献   

13.
The role of fimbria in adherence of an avian pathogenic Escherichia coli (APEC) O78 strain 789 to chicken intestine was studied. Bacterial adhesion to tissue sections representing the regions within the chicken intestinal tract was determined by using immunohistochemical methods. E. coli 789 grown to express the type 1 fimbria adhered efficiently to the crop epithelium, to the lamina propria of intestinal villi, and to the apical surfaces of both the mature as well as the crypt-located enterocytes in intestinal villi, whereas no adhesion to mucus-producing goblet cells was detected. The adhesion was inhibited by mannoside and the role of type 1 fimbriae in the observed adhesion was confirmed with a recombinant strain expressing type 1 fimbriae genes cloned from E. coli and Salmonella enterica. E. coli 789 strain grown to favor AC/I fimbriae expression as well as the recombinant E. coli strain expressing the fac genes adhered to goblet cells but only poorly to the other epithelial sites. E. coli strain 789 as well as S. enterica serovar Typhimurium IR715 and S. enterica serovar Enteriditis TN2 strains were able to multiply in ileal mucus medium. The type 1 fimbria expressing bacteria adhered to the ileal mucus, whereas the AC/I fimbriated strains showed poor adherence to the mucus. The adhesion of E. coli 789 onto the crop epithelium and the follicle associated epithelium of the chicken ileum was efficiently inhibited by an adhesive strain ST1 of Lactobacillus crispatus isolated from chicken, whereas poor inhibition of E. coli adherence was observed with the weakly adhesive L. crispatus strain 134mi. The type 1 fimbriae may be important in colonization of the chicken intestine by APEC and Salmonella.  相似文献   

14.
An avirulent, wild-type avian Escherichia coli (E. coli Av) was electrotransformed with a plasmid coding for the production of microcin 24 (pGOB18) and was designated E. coli AvGOB18. The transformant inhibited the growth of seven serotypes of Salmonella commonly associated with colonization and contamination of poultry products and seven strains of E. coli O157:H7 in the in vitro colicin/microcin assay. The transformant did not inhibit the replication of multiple isolates of Listeria monocytogenes or Campylobacter jejuni in similar assays. The transformant is nonconjugative, indicating that the plasmid would not be transmitted to other intestinal microflora in the environment. The transformant also survived in sterile tap and deionized water incubated at 25 C and 37 C in the laboratory for 30 days and was recovered from drinkers and birds in in vivo floor pen studies. In in vivo studies, E. coli AvGOB18 did not colonize the intestinal tract of broiler chicks when given as a single or multiple dose and did not reduce the Salmonella load in the broilers. But Salmonella typhimurium was reduced significantly in the intestinal tracts of broiler chickens when E. coli AvGOB18 was administered continually in the water supply.  相似文献   

15.
60株大肠杆菌的分离与致病性鉴定   总被引:1,自引:1,他引:0  
大肠杆菌(Escherichia coli,E.coli)是鸡肠道常在菌,部分菌株具有致病性。鸡大肠杆菌病就是由大肠杆菌中的某些致病性菌株引起的鸡感染性疾病。致病性大肠杆菌均携带毒力岛基因ChuA。为鉴定分离自临床病例的鸡大肠杆菌的致病性,本试验采用PCR法检测ChuA基因进行分子生物学鉴定,60株大肠杆菌中有31株属于致病性大肠杆菌,总阳性率为51.67%。  相似文献   

16.
鸡肠道菌基因组DNA的RAPD分类的研究   总被引:7,自引:0,他引:7  
本文以7种鸡肠道菌为研究对象,分别提取基因组DNA,利用随机扩增多态性DNA(RAPD)技术对其基因组进行DNA分析,筛选出一系列随机引物,能在7种肠道细菌中有较好的多态性,可用作分子标记鉴别这7种肠道菌。  相似文献   

17.
In the study 2004/2005, the current quantitative resistance level of Escherichia coli, Pasteurella multocida, Pseudomonas aeruginosa, Salmonella sp. and Staphylococcus aureus from chickens and turkeys was determined for the first time within the framework of the National Resistance Monitoring of the Federal Office of Consumer Protection and Food Safety (BVL).The objective was to implement a valid database on the basis of which the development and spread of resistance can be evaluated and monitored. During the investigation period from January 2004 to February 2005,927 strains were collected and 857 (92%) bacteria strains which corresponded to the specifications of the study protocol were tested with the broth microdilution method to determine the in vitro susceptibility (minimum inhibitory concentration) to 22 to 28 antimicrobial agents or antibiotic combinations. The results document a prevalence of resistance that exceeds that of bacterial pathogens of other animal species, especially in the case of tetracycline. Apart for S. aureus, clinical resistance to fluoroquinolones can still be considered low in poultry pathogens (E. coli approx. 2%). By applying the MICG of 4 mg/L for enrofloxacin, a susceptibility of approximately 78 % was calculated for S. aureus. A comparison of the prevalence of resistance between chickens and turkeys, showed that a slightly higher prevalence of resistance can be expected in turkeys. Differences between the susceptibility data of chicks and adult animals could only be found in turkeys. In the case of E. coli, the prevalence of resistance of strains isolated from adult turkeys was up to 10% higher than those isolated from chicks for the corresponding antimicrobial agents. It must be pointed out that the number of E. coli strains from adult turkeys was much higher (n = 194) than the number from turkey chicks (n = 21). The results indicate clearly that in a resistance monitoring system it is necessary to categorise poultry by animal species (chicken, turkey) as well as by production stage and type (broiler, laying hen), so that the epidemiology of resistance can be correctly represented and evaluated. This information is the basis for the development of long-term management options for minimizing antibiotic resistance. Furthermore, the knowledge of prevailing resistance levels in Germany is a valuable tool for veterinary practitioners when determining an empirical therapy.The data collected by the BVL make an important contribution to the optimisation of the safety of food from animals, and thus to improving consumer protection.  相似文献   

18.
M Dho  J P Lafont 《Avian diseases》1982,26(4):787-797
In chickens, virulent Escherichia coli strains express their pathogenicity in the respiratory tract. A quantitative comparison of tracheal colonization by virulent and avirulent E. coli was carried out in gnotoxenic chickens after intestinal implantation. Two-week-old axenic chicks reared in isolators were inoculated per os with various associations of identified E. coli strains. No clinical sign of disease was observed in any of the chicks, despite the presence of virulent strains in all the intestines and most of the tracheas. The virulent organism reached greater population sizes in the trachea and feces of monocontaminated chicks and of chicks contaminated simultaneously with a virulent and an avirulent strain. In holoxenic chicks, identified virulent and avirulent strains were outnumbered by the E. coli population of the intestinal flora previously established and could not be recovered from the tracheas of most chicks.  相似文献   

19.
Ultrasonic inactivation of Escherichia coli followed by irradiation was found to be the most efficient method for preparation of an effective vaccine against colibacillosis. Challenge experiments revealed that this vaccine provided the best protection compared with other methods of inactivation: heat, formaldehyde, and irradiation. Preparing the ultrasonicated vaccine from O2:K1 strain increased its range and also supported adequate protection against homologous strain O78:K80. The degree of protection conferred by the vaccine was positively correlated with the antibody titer against E. coli as measured on day of challenge. Low antibody titers detected 5 days post-vaccination resulted in only 20% protection. High antibody titers detected at 8 and 15 days post-vaccination correlated with a low number of chicks with lesions. In each challenged group, the live chicks that did not develop lesions had higher antibody titers than chicks with lesions, revealing a correlation between numbers of chicks with lesions and antibody titers as measured by enzyme-linked immunosorbent assay.  相似文献   

20.
Egg-yolk antibodies induced by immunizing hens with selected Escherichia coli antigens were evaluated for their ability to protect broiler chickens against respiratory/septicemic disease caused by avian pathogenic E. coli (APEC). Seven groups of broiler breeder hens were vaccinated three times, 1 week apart with live E. coli, killed E. coli, E. coli antigens [lipopolysaccharide (LPS), type 1 pilus adhesin (FimH), P pilus adhesin (PapG), aerobactin outer membrane receptor (IutA)] or phosphate buffered saline (PBS). An O78 APEC strain was used for preparation of all the antigens. Egg yolk immunoglobulins (IgY) were purified from eggs of each group and antibody activity in serum and purified IgY was determined by enzyme-linked immunosorbent assay (ELISA). IgY (100mg) was injected intramuscularly into 11-day-old broiler chickens, which were challenged 3 days later with homologous (O78) or heterologous (O1 or O2) E. coli by the intra-air sac route. Mortality was recorded and surviving chickens were euthanized 1 week after the challenge and examined for macroscopic lesions. Passive antibodies against all antigens except FimH were protective (90-100%) against the homologous challenge, but only anti-PapG and anti-IutA were effective against heterologous challenge. Anti-PapG IgY provided the greatest protection against the three serogroups of E. coli used for challenge. Hence vaccination of broiler breeders to induce anti-PapG and anti-IutA antibodies may provide passive protection of progeny chicks against respiratory/septicemic disease caused by APEC.  相似文献   

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