Investigation of the efficacy of a genetically-stabile live Salmonella typhimurium vaccine for use in swine

Hybrid swine (Landrace x Pietrain) aged 3-4 weeks were immunized twice at an interval of 3 weeks solely by the oral route and by the oral/parenteral route to evaluate the efficacy of a live S. Typhimurium vaccine. In each experiment a control group was run without vaccination. The animals were challenged at the age of 8-10 weeks by oral test infection with a labelled S. Typhimurium DT 104 strain. An ELISA was used to establish the presence of antibodies to S. Typhimurium in serum samples, coupled with clinical investigation. The presence of the challenge strain in the ileal and caecal mucosa and in the ileocolic lymph nodes was investigated quantitatively using the Koch plating method to determine the degree of colonization of those organs at the time of slaughter. The clinical course of disease was used to assess the success of vaccination. However, it was not possible to trigger, in a reproducible manner, clinical signs of disease in unvaccinated animals through infection. The vaccinated animals had a significantly lower (p < 0.05) colonization of the ileal and caecal mucosa than the unvaccinated animals. This was also seen to a lesser degree for the ileocolic lymph nodes.     

Use of a gonadotrophin-releasing hormone vaccine in headshaking horses

The aim of this study was to investigate the use of a gonadotrophin-releasing hormone (GnRH) vaccine in the treatment of headshaking in horses. Fifteen geldings received two doses of the GnRH vaccine four weeks apart. Serum was collected before and after vaccination to measure concentrations of luteinising hormone (LH) (10 horses) and follicle-stimulating hormone (FSH) (six horses). Owners recorded the frequency of seven common headshaking behaviours using a visual analogue scale (VAS) before vaccination and at two, four, eight, 12, 16 and 20 weeks after vaccination. Serum LH (P=0.008) and FSH (P=0.03) concentrations decreased significantly following vaccination. Although approximately one-third of the owners reported a subjective improvement in headshaking, serial scoring did not indicate a reduction in headshaking behaviours following vaccination with a commercial GnRH vaccine. Vaccination reactions were observed in four of 15 horses (27 per cent), including one case of severe, presumed immune-mediated, myositis.     

Salmonella flagellin enhances mucosal immunity of avian influenza vaccine in chickens

Flagellin, a bioactive Toll-like receptor (TLR) 5 ligand, may trigger the innate immunity that in turn is important for subsequent adaptive immune responses. In the present study, the adjuvant effects of the monomeric and polymeric forms of Salmonella flagellin (mFliC and pFliC, respectively) were examined in specific-pathogen free (SPF) chickens immunized intramuscularly (i.m.) or intranasally (i.n.) with formalin-inactivated avian influenza virus (AIV) H5N2 vaccines. Results showed that mFliC cooperating with the 64CpG adjuvant significantly induced influenza-specific antibody titers of plasma IgA in the i.m.-vaccinated animals. The nasal IgA levels in the i.n.-mFliC-coadministrated AIV vaccinated chickens were significantly elevated compared to levels observed in the control group (H5N2 vaccine alone). The pFliC cooperating with the 64CpG adjuvant significantly enhanced cell proliferation of splenocytes in the i.m.-vaccinated animals. TLR3 and TLR5 expressions were activated by flagellin stimulation in vitro and in vivo. These results suggest that flagellin can be used as an adjuvant in an AIV H5N2 vaccine, especially for mucosal immunity.     

Testing a bivalent vaccine in sheep against experimental infection with Salmonella bovismorbificans and Salmonella typhimurium

Abstract

Extract

Increasing awareness of the threat from salmonellosis to animal and human health, and recognition of implications to the farming economy and meat marketing industry in New Zealand, have emphasized the need for effective measures for controlling this disease. As pointed out by Beckett (1967 Beckett, F. W. 1967. The use of Salmonella vaccine in outbreaks of salmonellosis in sheep. N.Z. vet. J., 15: 66–69. [Taylor &; Francis Online] , [Google Scholar]), reduction in stocking rates or use of chemotherapeutic agents is often impracticable or ineffective. Other recent publications, drawing attention to the possible role of vaccination in controlling salmonellosis, have dealt with fundamental aspects of immunity (Jonas 1967 a Jonas, W. E. 1967a. Studies of the immunological aspects of salmonellosis of mice: active immunity. N.Z. vet. J., 15: 27–30. [Taylor &; Francis Online] , [Google Scholar]; b Jonas, W. E. 1967b. Salmonellosis of sheep: use of serum from a vaccinated ewe in a mouse passive protection test. N.Z. vet. J., 15: 55–61. [Taylor &; Francis Online] , [Google Scholar]) and with evaluation of an experimental vaccine under field conditions (Wallace and Murch, 1967 Wallace, G. V. and Murch, O. 1967. Field trials to assess the value of a bivalent killed salmonella vaccine in the control of ovine salmonellosis. N.Z. vet. J., 15: 62–65. [Taylor &; Francis Online] , [Google Scholar]).     

Evaluation of efficacy of a new live Salmonella Typhimurium vaccine candidate in a murine model

Efficacy of a new live Salmonella Typhimurium vaccine candidate attenuated by two virulence gene deletions was evaluated in this study. Live form of the vaccine was orally administered and formalin-inactivated form was intramuscularly (IM) administered. 105 female BALB/c mice were used and divided into 5 groups, A to E, containing 21 mice per group. Serum IgG and secretory IgA titers and levels of serum IFN-γ, IL-4, TNF-α, IL-12 of the immunized groups, especially group C (oral prime-oral booster) significantly increased. In addition, all animals in groups C showed no clinical symptoms and survived the virulent challenges, whereas all or some of mice from the other groups died of the challenge. These indicate that the vaccine candidate can be an effective tool for prevention of Salmonella infections by inducing robustly protective immune responses and leading to the production of inflammatory cytokines. Booster immunization, especially via oral administration, is necessary to optimize its protection.     

携带新城疫病毒F基因DNA疫苗的减毒鼠伤寒沙门氏菌的构建及其免疫原性

根据GenBank中发表的新城疫病毒(NDV)融合蛋白(F)基因序列,设计1对引物,通过RT-PCR扩增出鹅源NDV分离株JS5F基因(约1700bp),测序确认后,将其克隆入真核表达载体pVAX1,获得重组真核表达质粒pVAX1-F。pVAX1-F经脂质体转染COS-7细胞,间接免疫荧光试验检测出F基因在COS-7细胞中的表达产物。将pVAX1-F转化减毒鼠伤寒沙门氏菌SL7207,构建成功携带DNA疫苗的重组沙门氏菌SL7207(pVAX1-F)。重组菌以109CFU/只的剂量2次免疫BALB/c小鼠,免疫小鼠可以检测到特异性针对NDVF蛋白的血清抗体和小肠粘膜抗体应答,SL7207(pVAX1-F)免疫组抗体水平显著高于SL7207(pVAX1)组(P<0.05)。将SL7207(pVAX1-F)以109CFU/只剂量口服免疫1日龄雏鸡,免疫保护试验结果显示,SL7207(pVAX1-F)免疫组对鸡具有良好的保护率(77.27%),与空白对照组和SL7207(pVAX1)空载体组之间存在显著性差异(P<0.05)。结果表明,该运送DNA疫苗的减毒沙门氏菌系统在体内能成功释放所携带的质粒,并能刺激机体产生免疫应答,可对NDV强毒攻击提供良好的免疫保护作用,提示该疫苗候选株对新城疫的控制有重要应用前景。     

Vaccination of calves against Salmonella dublin with aromatic-dependent Salmonella typhimurium

Ten Holstein calves were divided into 2 groups. Five calves served as nonvaccinated controls, and 5 calves were vaccinated IM at 2 and 3 weeks of age with 10(9) aromatic-dependent (aro-) Salmonella typhimurium strain SL1479 containing O antigens 1, 4, 12. Serious adverse reactions to vaccination were not observed in the calves. Mean maximum rectal temperature increase in the vaccinated calves was 1.5 C. One calf had diarrhea and depressed appetite for 1 day after vaccination. At 5 weeks of age, all calves were challenge exposed orally with 1.5 X 10(11) virulent S dublin strain SL1367 (O antigens 9,12). After challenge-exposure inoculum was given, 1 of 5 vaccinated calves died and 4 of the 5 nonvaccinated calves died (P less than 0.05). Thus, some cross serotype protection against S dublin was induced by parenteral vaccination of calves with aro- S typhimurium strain SL1479, although protection was not complete.     

Percloacal infection of chickens with Salmonella typhimurium

The characteristics of Salmonella typhimurium infection in hatchlings and adult poultry given by the intravenous, oral and percloacal routes were compared. Based on LD₅₀ determinations, 1-day-old birds were most susceptible to percloacal infection. It is claimed that this infection technique yields predictable results in terms of mortality and numbers of organisms in the liver, spleen and intestinal tissues. Though older birds do not die when given large doses of organisms by this route, small numbers are sufficient to establish transient infection or carriage in the intestine.Based on quantitative data, the course of the percloacal infection appeared similar to that given by the oral route in hatchlings; however, the number of organisms required to establish infection by the former route was significantly less. It is suggested that the percloacal infection route may be a useful model for studying Salmonella infection-immunity in birds, particularly if resistance mechanisms operating in the intestine are to be taken into account. The possibility that it may be an important infection route in natural circumstances must also be considered.     

Evaluation of the live vaccine efficacy of virulence plasmid-cured,and phoP- or aroA-deficient Salmonella enterica serovar Typhimurium in mice

We evaluated the protective efficacy of 94-kb virulence plasmid-cured, and phoP- or aroA-deficient strains of Salmonella enterica serovar Typhimurium (ΔphoP or ΔaroA S. Typhimurium) as oral vaccine candidates in BALB/c mice. Two weeks after the completion of 3 oral immunizations with 1 × 10⁸ colony-forming units (CFU) of virulence plasmid-cured, and ΔphoP or ΔaroA S. Typhimurium at 10-day intervals, S. Typhimurium lipopolysaccharide (LPS)-specific mucosal secretory immunoglobulin A (s-IgA) antibody titers were detected in the cecal homogenate, bile and lung lavage fluid, but not in the intestinal lavage fluid. In addition, the increases in S. Typhimurium LPS-specific immunoglobulin G (IgG) and IgA antibody titers in the serum were also observed 2 weeks after completing 3 oral immunizations with virulence plasmid-cured, and ΔphoP or ΔaroA S. Typhimurium. The series of 3 oral immunizations protected the mice against an oral challenge with 5 × 10⁸ CFU of the virulent strain of S. Typhimurium, suggesting that both the virulence plasmid-cured, and ΔphoP and ΔaroA S. Typhimurium strains are promising candidates for safe and effective live S. Typhimurium vaccines.     

Use of a live attenuated Salmonella typhimurium vaccine to protect hens against Salmonella enteritidis infection while undergoing molt

Previous studies demonstrated that Salmonella enteritidis infections in hens undergoing molt via feed withdrawal were more severe than in full fed hens. We conducted two trials to determine if immunizing specific-pathogen-free, Salmonella-culture-negative hens via aerosol exposure to MeganVacl, a commercially available attenuated Salmonella typhimurium vaccine, would reduce transmission of S. enteritidis from infected hens to uninfected but contact-exposed hens during a molt. In trial 1, one group of hens received two aerosol doses of vaccine 2 wk apart whereas a second group of hens remained nonvaccinated. In trial 2, the vaccinated group received only one dose of vaccine. Two weeks after the final immunization, feed was removed from all the hens, and on day 4, the center hen in rows of 11 hens received a dose of 3 x 10(5) (trial 1) or 1.3 x 10(6) (trial 2). Transmission to the unchallenged hens was followed 3, 10, 17, and 24 days later. Vaccination reduced the horizontal spread of S. enteritidis in vaccinated hens compared with their nonvaccinated counterparts, with vaccinated hens shedding significantly less S. enteritidis on day 10 postchallenge in trial 1 and on days 3, 10, 17, and 24 in trial 2. Recovery of S. enteritidis from ovaries was significantly reduced in the vaccinated hens in trial 1 and from livers/spleens, ovaries, and cecum in trial 2. These studies indicate that immunization of hens with a live S. typhimurium vaccine could help reduce S. enteritidis problems during a molt situation.     

Immunogenicity of a DNA vaccine of Avian Reovirus orally delivered by attenuated Salmonella typhimurium

This study aimed to assess the ability of a Salmonella typhimurium-mediated Avain Reovirus DNA vaccine in eliciting antibody production. Six-day-old SPF chickens were orally immunized with SL7207 (pVAX-σC) twice at 2-week interval, detectable antibody was generated 2 weeks after immunization and was significantly higher than the control groups (P<0.01) and ten chickens (66.7%) were considered safe in the subsequent challenge. These results show that SL7207 (pVAX-σC) can induce protective antibody in chickens and the newly-constructed vaccine is also effective in protection chickens against ARV infection.     

Immunogenicity of a DNA vaccine of Avian Reovirus orally delivered by attenuated Salmonella typhimurium

This study aimed to assess the ability of a Salmonella typhimurium-mediated Avain Reovirus DNA vaccine in eliciting antibody production. Six-day-old SPF chickens were orally immunized with SL7207 (pVAX-σC) twice at 2-week interval, detectable antibody was generated 2 weeks after immunization and was significantly higher than the control groups (P < 0.01) and ten chickens (66.7%) were considered safe in the subsequent challenge. These results show that SL7207 (pVAX-σC) can induce protective antibody in chickens and the newly-constructed vaccine is also effective in protection chickens against ARV infection.     

Immunization of sows in an integrated pig-breeding herd using a homologous inactivated Salmonella vaccine decreases the prevalence of Salmonella typhimurium infection in the offspring

The efficacy of a homologous vaccination in preventing infection of suckling piglets with Salmonella (S.) Typhimurium was evaluated after an immunization of pregnant sows using an inactivated herd-specific S. Typhimurium vaccine. Twenty-five pregnant sows were vaccinated three times antepartum. The efficiency of this vaccine regime was assessed by comparison with a control group of 37 sows and their suckling piglets, which were daily treated with enrofloxacin from day 14 antepartum until the day of weaning. From the first day of life until day 142 post-partum, faecal samples of the piglets were collected and analysed for Salmonella shedding. In parallel, systemic antibody responses were monitored using a whole cell-based isotype-specific enzyme-linked immunosorbent assay (ELISA). The bacteriological investigation showed marked effects of vaccination. Salmonella Typhimurium could not be detected in any of the faecal samples of the piglets from the vaccinated sows. In contrast, the piglets of the group with long-time antibiotic treatment shed salmonellae rating to 47.4% of the animals. Furthermore, the offspring from vaccinated sows showed significantly decreased antibody activities of immunoglobulin (Ig)A and IgG. These bacteriological and serological results indicate a significantly lower Salmonella prevalence in piglets of the vaccinated group. As this study shows, the presented strategy of vaccination of pregnant sows with an inactivated Salmonella vaccine seems to be a suitable measure in decreasing Salmonella prevalence in offspring of infected sows.     

Expression of Escherichia coli antigens in Salmonella typhimurium as a vaccine to prevent airsacculitis in chickens

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.