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1.
A virulent Salmonella dublin isolate was made histidine-requiring (his-) to allow recognition. The his- derivative, SL1367 (still calf-virulent), was then given by transduction and mutation, a transposon-generated non-reverting aromatic biosynthesis (aro) defect; this defect caused loss of virulence for the mouse. The his- aro- derivative strain, SL1438, was effective as a live vaccine in mice. Twenty male Holstein calves were divided into 4 groups. Groups I, II, and III were vaccinated IM at 2 weeks and at 3 weeks of age with aromatic-dependent (aro-) S dublin strain SL1438. Groups I and III received freshly prepared vaccine and group II received lyophilized vaccine. Serious adverse reactions to the vaccination were not seen. After vaccination, the mean maximum increase in rectal temperature was 1.8 C in group I and III calves and 0.6 C in group II calves. Fewer group II calves developed diarrhea (1 of 5) or positive blood cultures (0 of 5) after vaccination compared with group I and III calves (6 of 10 and 5 of 10, respectively). Postvaccination diarrhea was mild and of short duration. Group IV was comprised of 5 nonvaccinated calves. At 5 weeks of age, all calves were challenge exposed orally. Group I, II, and IV calves were challenge exposed with 10(11) virulent S dublin SL1367. Group III was challenge exposed with 10(11) virulent S typhimurium UCD 108-11. Subsequently, fever and diarrhea (lasting 1 to 3 days), but no deaths, were observed in the vaccinated calves. Four of the 5 nonvaccinated (group IV) calves died (P less than 0.001) within 8 days after challenge exposure. Aro- S dublin SL 1438 did not cause serious adverse effects and provided protection against oral challenge exposure with either virulent S dublin or S typhimurium.  相似文献   

2.
Migration of bovine macrophages under agarose was used to assess cellular immunity in 7 nonvaccinated calves and 9 calves vaccinated with Salmonella typhimurium. The 9 vaccinated calves were allotted to 4 groups. Group I calves were vaccinated twice orally with small doses of virulent S typhimurium; group II calves were vaccinated twice orally with genetically altered aromatic-dependent (aro-) S typhimurium SL3261; group III calves were vaccinated twice IM with small doses of virulent S typhimurium; and group IV calves were vaccinated twice IM with aro- S typhimurium SL1479. Samples of blood were obtained from these calves at 2 weeks after the 2nd vaccinal dose was given, and lymphocytes were harvested, using lymphocyte separation medium. Lymphocytes in serum-free medium were then incubated with S typhimurim antigen for 48 hours. Lymphocytes were then transferred to antigen-free medium and incubated for 48 hours, and the supernatant was assayed for the migration-inhibition factor (MIF). Lymphocyte supernatant was assayed for MIF by incubating it for 48 hours with 2.0 X 10(4) alveolar macrophages in agar wells. The macrophage migration distance was measured and compared with control values. Macrophage migration was inhibited in the presence of supernatant of lymphocytes from vaccinated calves that had been incubated with antigen, indicating the presence of the MIF in the supernatant. Migration distances, as a percentage of control, were 33% for group I calves (oral vaccination, virulent vaccinal organism), 60% for group II calves (oral vaccination, aro- vaccinal organism), 41% for group III (IM vaccination, virulent organism), and 25% for group IV (IM vaccination, aro- vaccinal organism).(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

3.
Aromatic-dependent Salmonella typhimurium as modified live vaccines for calves   总被引:29,自引:0,他引:29  
Strains of Salmonella sp with complete nonreverting aromatic biosynthesis (aro) defects are expected to be nonvirulent, in respect to invasive infection, because they need the aromatic metabolites paraaminobenzoate (for making folate) and dihydroxybenzoate (for making enterochelin) which are not available in host tissues. Derivatives with transposon-generated complete nonreverting aro-defects were prepared from 3 mouse-virulent strains of S typhimurium, namely, FIRN, WRAY, and UCD. The latter 2 parent strains originally were isolated from calves and are known to be calf-virulent. The resultant aromatic-dependent (aro-) strains were used to vaccinate 27 calves (2 to 3 weeks old), usually giving 2 doses by the IM route (10(9) bacteria) or orally (1.5 X 10(11)). Vaccination did not cause severe ill effects in any calf. Thus aro- defects cause loss of virulence for calves, as previously shown for mice. Vaccinated and control calves were challenge exposed, usually at 5 weeks of age, by feeding 1.5 X 10(11) cells of 1 of 2 calf-virulent S typhimurium strains, either UCD 108-11 or SL1323. Of the 16 challenge-exposed control calves, all became anorectic and depressed (CNS), and 15 had diarrhea. Fourteen of the 16 died; all tested tissues were bacteriologically culture-positive for Salmonella at necropsy. Vaccination with the live UCD aro- vaccine strain, SL1479 by either of 2 schedules (IM or orally) appeared effective.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

4.
Mice immunized with an aromatic-dependent (aro-) S. dublin strain CS101 by either the intraperitoneal (i.p.) or oral route, were protected against oral challenge with a virulent S. dublin strain CS90, the degree of protection being the greatest when mice had received 3 immunizing doses at weekly intervals. Mice immunized with an aromatic-dependent (aro-) S. typhimurium strain CS332 by the i.p. or oral routes were protected against challenge with virulent S. dublin strain CS90 at 1 or 2 weeks but not at 3 or 4 weeks post-immunization. Mice immunized with 1 dose of aro- S. dublin strain CS101 by the i.p. route developed low levels of lipopolysaccharide (LPS) and flagellin-specific antibody but no delayed-type hypersensitivity (DTH) whereas those immunized with 2 or 3 doses developed significantly higher antibody titres and DTH. In contrast, mice immunized by the oral route developed neither significant antibody response nor DTH. The aro- S. dublin strain CS101 could not be detected beyond day 28 post-inoculation in visceral organs including liver, spleen, mesentery, small intestine, caecum or large intestine of mice inoculated by the i.p. route or in mice inoculated by the oral route with the exception of day 42 post-inoculation. Challenge of mice previously immunized with 3 doses of the aro- S. dublin strain CS101 by the i.p. or oral route with virulent S. dublin strain CS90 resulted in their rapid clearance from the above visceral organs. Calves immunized with the aro- S. dublin strain CS101 by either the intramuscular (i.m.) or oral routes were significantly protected against oral challenge with virulent S. dublin strain CS90. In contrast to the observations in mice, somatic (O) and flagellar (H) antibody titres of calves immunized by either route were negligible as were anti-LPS antibody titres. However, flagellin-specific antibody titres were higher in calves immunized by the i.m. than the oral route. These results indicate that the protection observed in immunized mice or calves against oral challenge with virulent S. dublin was unlikely to have been mediated by humoral salmonella-specific immune mechanism(s).  相似文献   

5.
Eight calves were immunized with live auxotrophic Salmonella typhimurium mutants (aro -SL 1479, gal E 3821) and twelve calves with phenol-killed whole-cell S. typhimurium vaccine, respectively. The clinical status of the animals was followed and serial reisolation of vaccine and challenge strains from faeces was attempted. The immunization of calves with the live aro- auxotrophic S. typhimurium SL 1479 mutant proved to be unsuitable due to the death of calves after revaccination. The calves immunized with live auxotrophic gal E S. typhimurium CCM 3821 mutant proved to be protected against challenge with virulent S. typhimurium 4/5 strain administered orally at a dose of 10(6) colony forming units (CFU). The postvaccination complications showed serious shortcomings. The immunization of calves with three doses of whole-cell inactivated vaccine containing 5 strains of S. typhimurium was effective against oral challenge with virulent S. typhimurium 4/5 at a dose of 10(6) CFU.  相似文献   

6.
Twenty-six clinically normal colostrum-fed dairy calves were allotted to 5 groups. Calves of groups 1 and 2 served as nonvaccinated controls and were challenge-exposed with variable numbers of organisms. Group-3 calves were vaccinated SC with a modified Salmonella typhimurium bacterin. The bacterin was composed of killed acid-hydrolyzed S typhimurium G30/C21 (Re-mutant) whole cells coated with alkali-hydrolyzed S typhimurium LT-2 lipopolysaccharide, as antigen, and monophosphoryl lipid A, as adjuvant. Calves of groups 4 and 5 were vaccinated with a 2% mineral oil-in-water emulsion containing lipopolysaccharide as antigen and monophosphoryl lipid A and trehalose 6-6'-dimycolate as adjuvants. Calves of groups 3-5 were vaccinated at 2 weeks of age and again at 4 or 6 weeks of age. Adverse reactions were not observed after vaccination. Calves were challenge-exposed orally at 6 or 8 weeks of age with 1.5 X 10(11) (groups 1 and 4), or 3.0 X 10(11) (groups 2, 3, and 5) colony-forming units of S typhimurium UCD 108-11. Mortality after challenge exposure was 2 of 5 group-1 calves; 4 of 5 group-2 calves; 5 of 6 group-3 calves; 1 of 5 group-4 calves; and 4 of 5 group-5 calves. Statistical difference between calves of similarly challenge-exposed groups was not evident, indicating failure of either vaccine to protect calves of this age from oral challenge exposure with virulent S typhimurium.  相似文献   

7.
The purpose of this study was to evaluate the safety and efficacy of a diaminopimelic acid mutant of Salmonella typhimurium as a vaccine for calves. Transposon techniques were used to create a stable nonreverting diaminopimelic acid mutant of a virulent S. typhimurium strain. Calves were vaccinated at weekly intervals with the diaminopimelic acid mutant given as an oral dose of 10(10) organisms, followed by two subcutaneous doses of 10(9) organisms. The calves tolerated vaccination well and the vaccine strain was eliminated from the feces within four days. Of five vaccinated calves, three survived challenge with 5 X 10(9) organisms of the parent strain whereas all five unvaccinated calves that were challenged died. The surviving calves eliminated the challenge organism from the feces within three weeks.  相似文献   

8.
Chemiluminescence of bovine alveolar macrophages was used to study the development of opsonins in calves vaccinated parenterally with live aromatic-dependent strains of either S. dublin or S. typhimurium. These calves responded by producing Salmonella-specific opsonins detected by increased chemiluminescent responses, and were able to survive oral challenge with live virulent organisms of either serotype. Non-vaccinated calves of the same age lacked Salmonella-specific opsonins and were not able to survive challenge. Thus it was concluded that the ability to produce opsonins is among the immunological responses that are associated with protection against salmonellosis in calves. Antigenic similarities between S. dublin and S. typhimurium were shown by the ability of either organism to absorb significant amounts of opsonic capacity from the sera of calves vaccinated with either of the two vaccines. These antigenic similarities are thought to explain in part the ability of either vaccine to protect against challenge with either the homologous or heterologous Salmonella serotype.  相似文献   

9.
The purpose of this study was to evaluate the effectiveness of an aromatic-dependent mutant of Salmonella typhimurium as a parenteral vaccine for prevention of fecal shedding of Salmonella spp. Pigs and chickens were vaccinated IM, with 1 x 10(9) and 1 x 10(8) organisms, respectively, followed by a second identical vaccination 2 weeks later. Salmonella organisms were not detected by analysis of fecal or cloacal swab specimens from any animal after vaccination. Deleterious side effects were not noticed after vaccination. Pigs were challenge-inoculated PO with 1 x 10(12) virulent S typhimurium 1 week after the second vaccination. Chickens were challenge-inoculated PO with 3 x 10(8) organisms of either S enteritidis or the virulent parent strain of S typhimurium 3 weeks after the second vaccination. Vaccinated pigs shed Salmonella spp significantly less frequently than did nonvaccinated pigs. Vaccinated chickens challenge-inoculated with either S enteritidis or S typhimurium also shed Salmonella less frequently than the corresponding nonvaccinated control birds; however, the difference was not significant.  相似文献   

10.
Salmonella dublin strain SL5631, which is auxotrophic for p-amino-benzoic acid and 2,3-dihydroxybenzoate because of a deleted aroA gene, was given orally in a dose of 10(10) live bacteria to 6 calves 5-7 weeks old. The calves tolerated the strain well, had a transient mucoid diarrhea and sacrificed animals showed a moderate acute inflammation in the ileum on day 2. The salmonella strain was seen lining the mucosal epithelium using immunohistopathology. Already in calves sacrificed on day 6 the damage was less pronounced and signs of regeneration were obvious. The healing process was more accentuated in calves sacrificed on day 14. The results demonstrated the attenuating effect of the deleted aroA gene. Groups of 5-7 weeks old calves (n = 25) orally immunized with 10(8), 10(9) and 10(10) S. dublin SL5631 at weekly intervals were challenged 2, 6 or 15 weeks after the immunization. All calves were protected against oral challenge with 10(10) bacteria of the virulent S. dublin strain, which equals 1,000 LD50 doses. At autopsy, calves were sacrificed 3 weeks after challenge, all calves had normal intestinal findings with only slightly enlarged mesenteric lymph nodes. The protective effect is surmised to involve cell-mediated as well as humoral defense mechanisms.  相似文献   

11.
The minimum number of doses of a live aromatic dependent (aro-) Salmonella typhimurium vaccine strain (SL1479), given by the intramuscular, oral or subcutaneous route required to protect sheep from experimentally-induced clinical salmonellosis, was determined. A significant reduction in mortalities and diarrhoea occurred in those sheep immunised with one or 2 intramuscular doses or 2 subcutaneous doses. On the other hand, sheep immunised with one subcutaneous dose were not protected. Immunisation with one or 2 oral doses also resulted in a significant reduction in mortality, although reduction in the prevalence of severe diarrhoea was less consistent. Sheep immunised with a single intramuscular dose of aro- S. typhimurium developed high levels of serum antibodies and significant delayed-type cutaneous hypersensitivity response to homologous Salmonella lipopolysaccharide and flagellin, whereas those with a single oral dose did not. It was concluded that immunisation of sheep with a single oral or intramuscular dose of live aro- S. typhimurium reduced mortalities and the prevalence of diarrhoea in sheep due to infection with virulent S. typhimurium.  相似文献   

12.
Cows were vaccinated with formalin-killed Salmonella typhimurium approximately seven weeks and two weeks before parturition to investigate whether passive immunity could protect their calves against experimental S typhimurium infection. After birth the calves were left with their dam for 48 hours and then separated and fed cold, stored colostrum from their own dam for a further eight days. Oral challenge five days after birth with 10(8) S typhimurium did not result in the death of these calves even when they had absorbed little colostrum. Mortality was reduced to 22 per cent in calves which sucked from vaccinated dams and were then fed colostrum from unvaccinated cows and to 50 per cent in calves born to unvaccinated cows and later fed colostrum from vaccinated animals. Calves which sucked from a vaccinated dam and then received stored colostrum from the same cow excreted salmonellas for significantly shorter periods after challenge and were less often infected at necropsy 28 days after inoculation. Protection was not correlated with the levels of O or H agglutinating antibodies in serum, which were at a maximum 24 hours after sucking and then slowly declined. There was no evidence of an active antibody response in the serum. Measurement of the O and H response of cows after vaccination indicated that the vaccination schedule could be improved. The highest levels of agglutinating antibody were measured between two and three weeks after the first vaccination and there was only a minimal response to the second vaccination before parturition.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

13.
A commercially available Salmonella bacterin was administered to Holstein calves starting at 1 to 19 weeks of age. Serum samples were obtained before administering bacterin and at 2-week intervals thereafter. An ELISA with Salmonella dublin lipopolysaccharide (LPS) or S dublin whole cells as antigen, was used to measure specific IgG and IgM responses. Antibody responses to LPS were not detected from calves < 12 weeks old inoculated with killed bacterin. Immunoglobulin responses to whole-cell antigen were detected from all age groups of calves inoculated with the same killed Salmonella bacterin. Calves < 11 weeks old are able to produce immunoglobulins to some whole-cell antigens, but are unable to produce anti-LPS immunoglobulins when inoculated with killed Salmonella bacterin. This age-related response to killed Salmonella antigens may account, in part, for increased susceptibility to salmonellosis in calves < 12 weeks old. In comparison to the response for killed antigen, 8 calves given modified-live aromatic-dependent S dublin bacterin at 1 to 3 weeks of age had detectable anti-LPS immunoglobulins after immunization, although the response was not as rapid and was of a lesser magnitude than that of older calves given killed Salmonella bacterin.  相似文献   

14.
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   

15.
Pregnant cows were vaccinated at ablactation by infusion of heat inactivated S. dublin or S. typhimurium into the mammary gland in order to protect their offsprings via colostrum against salmonellosis. This method of vaccination is based upon statements according to which specific IgA and IgM play a prominent role in respect of protection. Both Ig classes are produced locally and are not channelled from the mother's system into the mammary gland. In comparison to calves from non-vaccinated control animals, calves from vaccinated cows did not exhibit clinical symptoms after challenge infection, and had a reduced salmonella excretion rate in respect of quantity but not in respect of the duration of shedding salmonella. Serological quantitation of specific Ig-classes (DIG-ELISA) did not allow to identify a specific Ig-class responsible for protection.  相似文献   

16.
Previous findings, viz. that mice can be successfully immunized against infection with Salmonella dublin with either live or inactivated vaccine, were confirmed. Immunity lasted for at least 12 weeks in mice which had been immunized with inactivated alum-precipitated vaccine. The immunogenicity of inactivated vaccine gradually decreased on storage at 4 degrees C, but this was only detectable if a single injection was used for immunization: 2 injections virtually eliminated this phenomenon. The immunogenicity of live vaccine in mice was not enhanced by levamizole or the simultaneous injection of inactivated organisms. Both live and inactivated vaccines provided immunity in calves. A single injection of lyophilized vaccine, prepared from live rough Salmonella dublin strain (HB 1/17),protected 3 out of 6 calves, while 2 injections of a formalin-inactivated, alum-precipated vaccine, containing 1% packed cells of S. dublin strain 2652 V, protected 5 out of 6 calves against intraduodenal challenge with 2 x 10(9), S. dublin strain 2652 V. Two calves which had been immunized with an inactivated oil adjuvant vaccine were also solidly immune to this challenge. Serum antibody response in calves was poor when measured by the tube agglutination and the haemagglutination tests. Similarly, the sera had only marginal protective values when tested by means of a passive protection test in mice. Antibody titres alone are not a valid measure therefore, for the immune status of immunized animals.  相似文献   

17.
The aim of this study was to examine the duration of immunity of different vaccination schemes using the S. enteritidis live vaccine Gallivac Se and the S. enteritidis-S. typhimurium inactivated vaccine Gallimune Se+St. Three groups of Lohman Brown chickens were used. Group one was vaccinated three times orally with Gallivac Se at weeks one, seven and 13 of age. Group two was vaccinated twice orally with Gallivac Se in weeks one and seven and once i.m. with Gallimune Se+St in week 14 of age. A third group was not vaccinated and served as the control group. Eight randomly selected chickens from each of the three groups were challenged with a nalidixic acid resistant S. enteritidis PT4 strain in weeks 24, 51 and 71 of age and the same number of animals were challenged with a S. typhimurium DT 104 strain in weeks 26, 54 and 73 (75) of age.The chickens were euthanised seven days post challenge and the number of challenge strain organisms (log10 cfu) in the liver and on caecal mucosa was determined.The quantitative investigation of the challenge strain in the liver and caecal mucosa revealed a statistically significant (p < 0.05) lower challenge strain burden in the vaccinated groups compared with the non-vaccinated control group up to week 71 (73) of age. The protective effects were demonstrated for both challenge strains.  相似文献   

18.
An oil emulsion Escherichia coli bacterin administered in 1- and 2-dose vaccination regimens was evaluated in beef cattle. Serologic responses to the K99 pilus antigen were monitored, and suckling offspring from vaccinated and nonvaccinated cows were inoculated with virulent, K99-positive, enterotoxigenic Escherichia coli. The degree of protection and duration of immunity conferred were determined in 2 respective studies. In the first study (study A), titers of pregnant cattle were determined from time of vaccination through calving (a 6- to 20-week period). Titers of 24 cows vaccinated with a single 2-ml dose of bacterin were compared with those of 24 cows given a 2-dose regimen and with those of 23 nonvaccinated cattle (contemporary controls). Inoculum consisting of 1.2 X 10(12) viable enterotoxigenic E coli/dose administered to nursing calves from these dams yielded 0% mortality (0 deaths/20 calves) in calves from 1-dose vaccinates, 6% mortality (1 death/18 calves) in calves from 2-dose vaccinates, and 37% mortality (7 deaths/19 calves) in calves from nonvaccinated dams. Study B was an extended evaluation conducted in cattle that were kept in the study up to 87 weeks from initial vaccination until calving. Serologic titers to the K99 pilus antigen were compared in 1-dose, 2-dose, and nonvaccinated cattle in groups of 8, 6, and 6, respectively. Calves from these dams were inoculated with 8.1 X 10(11) viable enterotoxigenic E coli/dose, which resulted in 0% mortality (0 deaths/5 calves) in calves from 1-dose vaccinates, 0% mortality (0 deaths/5 calves) in calves from 2-dose vaccinates, and 80% mortality (5 deaths/6 calves) in calves from nonvaccinated dams.  相似文献   

19.
The purpose of the study was to evaluate the safety and efficacy of a galactose epimeraseless mutant of Salmonella typhimurium administered as an oral vaccine to one week old calves and to investigate properties of galactose epimeraseless mutants which affect their virulence and immunogenicity. The galactose epimeraseless mutant S. typhimurium strain G30D caused diarrhea and fever in three calves to which it was administered orally at a dose of 10(10) organisms; all three calves died following challenge with virulent S. typhimurium ten days postvaccination. Mild illness developed in four calves vaccinated with a dose of 9 X 10(6) organisms and one of these calves survived challenge. Three unvaccinated calves died following challenge. The vaccine organism persisted in tissues and was shed for a prolonged period by calves which received 10(10) organisms. Studies of characteristics of galactose epimeraseless mutants of S. typhimurium showed that, in the presence of galactose, there is selection for secondary mutants which are galactose resistant. The studies indicate that galactose epimeraseless mutants of S. typhimurium are not good candidate live vaccine organisms for use in calves.  相似文献   

20.
A controlled calfhood vaccination trial to prevent bovine virus diarrhea was conducted in a 100 head cow-calf operation with a three year history of annual calf losses due to enteric bovine virus diarrhea (persistently infected herd). Approximately 50% of the calves were vaccinated at six, 12 and 24 weeks of age. Paired serum samples and growth data were collected on three occasions for comparison between vaccinates and controls. Three vaccinated calves died of enteric bovine virus diarrhea in the first year of the trial and one nonvaccinated calf died in the second year. Two of the three vaccinated calves had developed bovine virus diarrhea virus neutralization antibody titres of 2048 or greater before developing clinical signs. The control and third vaccinated calf failed to seroconvert before dying of enteric bovine virus diarrhea. Approximately 90% of the vaccinated calves seroconverted compared to approximately 40% of the controls. Paired serum samples collected from 75% of the cows in the spring, summer and fall of each year of the trial, showed persistent high bovine virus diarrhea virus neutralization titres in all samples. Calf vaccination before 12 weeks of age had little effect on seroconversion due to high levels of passive antibody to bovine virus diarrhea. Growth data showed that there was no improvement in weight gain or rate of growth in the vaccinated calves.  相似文献   

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