Fowl cholera: influence of Bordetella avium on vaccinal immunity of turkeys to Pasteurella multocida

Turkeys exposed to Bordetella avium were vaccinated against fowl cholera with live Pasteurella multocida vaccine. Previous exposure to B. avium resulted in impairment of systemic immunity conferred by the vaccine: 86% of the vaccinated turkeys exposed to B. avium at 1 day old developed lesions or died of fowl cholera after challenge at 15 weeks old with virulent P. multocida. Of vaccinated turkeys not previously exposed to B. avium, only 26% had lesions or died of fowl cholera.     

Efficacy of a commercial turkey coryza vaccine (Art-Vax) in turkey poults

Four laboratory experiments were designed to study the efficacy of the only available commercial vaccine for turkey coryza, Art-Vax. Poults were vaccinated either once or twice at different ages and challenged with pathogenic Alcaligenes faecalis. In another study, commercial turkeys vaccinated at 1 and 12 days of age on a commercial farm were brought to the laboratory for challenge with pathogenic A. faecalis. Both the laboratory- and field-vaccinated poults were given the manufacturer's recommended dosage of the vaccine strain. Regardless of the vaccine schedule or source of poults, the vaccine was not effective in protecting challenged turkeys from infection. Furthermore, the vaccine was not effective in protecting poults less than 3 weeks of age from disease, but it was effective in protecting poults more than 3 weeks of age from disease. These results indicate that although vaccinated turkeys older than 3 weeks of age were not susceptible to disease, they were susceptible to infection and could act as carriers of field strains of A. faecalis, thus perpetuating the risk of infection to flocks subsequently raised in the same buildings.     

Protection of chickens by ribosomal vaccines from Pasteurella multocida: dependence on homologous lipopolysaccharide

Chickens were protected against fowl cholera by ribosomal vaccines prepared from noncapsulated Pasteurella multocida. Passive hemagglutination (PHA) titers to lipopolysaccharide (LPS) and the degree of protection conferred by ribosomal vaccines were diminished or abolished when ribosomes were chromatographed on an immunoadsorbent column. Addition of subimmunogenic amounts of serotype 1 (homologous) LPS to highly purified ribosomes resulted in vaccines that protected against challenge exposure and produced PHA titers to homologous LPS. Addition of serotype 5 LPS to highly purified ribosomes did not protect chickens against challenge exposure with serotype 1 P multocida, but produced PHA titers to serotype 5 LPS. Combinations of serotype 1 ribosomal RNA and serotype 1 (homologous) LPS did not protect chickens or produce PHA titers to LPS. Purified ribosomes from Brucella abortus, Aspergillus fumigatus, and chicken liver were combined with LPS from P multocida and were evaluated as vaccines. Brucella abortus and A fumigatus ribosomes combined with LPS protected chickens as well as did bacterin made from whole cells of P multocida. Chicken liver ribosomes combined with LPS did not provide protection. To determine whether a protein carrier would substitute for ribosomes, methylated bovine albumin (MBA) was combined with LPS and evaluated as a vaccine. A serologic response to LPS was induced by MBA-LPS vaccine, but the vaccine offered no better protection than when LPS was used alone as vaccine. Ribosome-LPS vaccines produced serologic responses to LPS that were at least 5-fold greater than those produced by MBA-LPS vaccine.     

Field vaccination against hemorrhagic enteritis of turkeys by a cell-culture live-virus vaccine

A cell-culture-propagated (CC) live-virus hemorrhagic enteritis (HE) vaccine was evaluated for efficacy and safety in two field trials conducted in North Carolina (NC) and Minnesota (MN). At 4 or 5 1/2 weeks of age, 9,839 poults in NC and 15,857 poults in MN were vaccinated with a CC HE vaccine administered via the drinking water. A comparable number of poults were maintained as unvaccinated controls. Vaccinated and unvaccinated poults were compared for seroconversion, response to laboratory challenge with a virulent HE virus at 3 weeks postvaccination, livability, percentage graded A, and average weight at marketing. In both trials, vaccination with the CC HE vaccine resulted in immunity against HE as indicated by seroconversion and by resistance to HE lesions following laboratory challenge with virulent HE virus. Compared with unvaccinated groups, vaccinated groups had a significantly higher percentage of turkeys graded A in the NC trial and in two of three flocks in the MN trial (P less than 0.005). Further, in the NC trial, livability was significantly higher (P less than 0.005) in vaccinated turkeys than in unvaccinated turkeys. These data indicate that the CC HE vaccine is efficacious and safe to use in the field.     

Protective efficacy of H5 inactivated vaccines in meat turkey poults after challenge with Egyptian variant highly pathogenic avian influenza H5N1 virus

In contrast to chickens, there is a paucity of information on the potency of H5 vaccines to protect turkeys against the highly pathogenic avian influenza (HPAI) H5N1 virus infections. In this study, 4 groups, 10 turkey poults each, were vaccinated at seven days old with one of H5N2 or H5N1 commercial vaccines or one of two prepared H5N1 vaccines from a local Egyptian variant HPAI H5N1 (EGYvar/H5N1) strain. At 35 days age, all vaccinated and 10 non vaccinated birds were challenged intranasal with 10(6) EID(50)/0.1 ml of EGYvar/H5N1. All vaccines used in this study were immunogenic in turkeys. There was no cross reaction between the commercial vaccines and the Egyptian variant H5N1 antigen as obtained by the hemagglutination inhibition test. Birds vaccinated with H5N2 vaccine were died, while other H5N1 vaccinated groups have had 20-40% mortality. The highest virus excretion was found in non-vaccinated infected and H5N2 vaccinated birds. Eleven peculiar amino acid substitutions in H5 protein of the variant strain were existed neither in the vaccine strains nor in the earliest H5N1 virus introduced into Egypt in 2006. In conclusion, single vaccination at seven days old is inadequate for protection of meat turkeys against variant HPAI H5N1 challenge and multi-dose vaccination at older age is recommended. For the foreseeable future, continuous evaluation of the current vaccines in H5N1 endemic countries in the face of virus evolution is a paramount challenge to mitigate the socio-economic impact of the virus.     

Immunomodular effect of fusion gene DNA vaccine of avian metapneumoviruses

The objective of this work was to develop and evaluate the immunomodular effect of a DNA vaccine based on the fusion (F) gene of avian metapneumoviruses (aMPV) and to study its protection against field virus challenge, as this will help to better control the disease in turkeys. In this study, the F protein of the Egyptian isolate (Giza-turkey rhinotracheitis-4) of the B-subtype of aMPV isolated in 2009 was expressed from a DNA plasmid in Vero cells. After 1 i.m. injection of turkey poults with this plasmid, the antibody response was detected by ELISA. The turkey poults inoculated with locally prepared DNA aMPV vaccine had highly significant phagocytic activity, as measured by phagocytic percent and index of macrophage activation, in comparison to those inoculated with inactivated and live attenuated vaccines and with the noninfected control group. Intratracheal challenge of turkey poults at 21 d postvaccination by a dose of 100 uL of field Egyptian Giza-turkey rhinotracheitis-4 virus of a titer 6 log₁₀ tissue culture infective dose 50 resulted in 100% protection in poults that received locally prepared DNA aMPV vaccine, whereas those that received commercial aMPV vaccines experienced 80 and 90% protection; typical clinical signs of aMPV infection were seen in control nonvaccinated poults. Therefore, a high success rate was noted when using F gene DNA plasmid vaccine by the induction of a potent immunomodular effect for both cell-mediated and humoral immune response. The use of the F gene DNA plasmid vaccine developed in this study provided 100% protection in vaccinated poults, which can help in controlling aMPV infections in turkeys.     

Effects of bursectomy, irradiation, and cyclophosphamide on turkeys vaccinated with CU cholera strain

Turkeys surgically bursectomized, irradiated, and/or injected with cyclophosphamide at 1 day were vaccinated with the live Clemson University (CU) strain of Pasteurella multocida. Bursectomized turkeys vaccinated via drinking water or wing-web puncture at 7 weeks and challenged at 11 weeks had a significantly (P less than 0.05) lower survival rate after challenge than unbursectomized controls. Bursectomized and unbursectomized turkeys vaccinated via drinking water at 7 weeks, revaccinated via the auditory tube at 11 weeks, and challenged at 15 weeks had similar survival rates. The vaccinated bursectomized turkeys had significantly (P less than 0.05) lower levels of serum anti-P. multocida antibody than vaccinated unbursectomized controls. Radiation had no immunosuppressive effect. The immunosuppressive effect of cyclophosphamide was dosage-dependent. Bursectomy and injection of cyclophosphamide in the same turkey were complementary. It was concluded that in young turkeys, the development of immunity to the avirulent CU vaccine is highly dependent upon the bursa of Fabricius, but that as they grow older the bursa is of less importance, particularly if they were vaccinated via a parenteral route, such as in the air spaces of the head.     

Serum and mucosal antibody responses and protection in pigs vaccinated against Mycoplasma hyopneumoniae with vaccines containing a denatured membrane antigen pool and adjuvant

Objective To investigate the protective efficacy of a pool of denatured membrane protein antigens of Mycoplasma hyopneumoniae (J strain) in the molecular size range 70 to 85 kDa (F3 antigen) in combination with adjuvants for pigs challenged with M hyopneumoniae .
Design A vaccine efficacy experiment with assessment of serum and respiratory tract antibody responses.
Procedure F3 antigens were emulsified with five different adjuvants. To groups of three pigs per vaccine, four vaccines were given by intramuscular injection, and two vaccines, including one of those given intramuscularly, were given by intraperitoneal injection.
Results Compared to six unvaccinated pigs, animals vaccinated with F3 antigen displayed significantly reduced pneumonia (54% reduction in mean lung score) following experimental challenge. Analysis of post-vaccination, pre-challenge IgG and IgA ELISA antibody absorbances in serum and respiratory tract washings revealed no correlation with lung score. Six weeks after challenge, pigs previously vaccinated intramuscularly mostly demonstrated greater IgG and IgA responses in respiratory tract washings, and greater IgG serum antibody responses, than those vaccinated by intraperitoneal injection.
Conclusion Pigs vaccinated with M hyopneumoniae antigens in the molecular size range of 70 to 85 kDa showed a significant reduction in lung lesions compared with unvaccinated control animals after experimental challenge. IgG and IgA antibody concentrations in serum and respiratory tract washings after vaccination do not provide a useful prognostic indicator of protection from enzootic pneumonia.     

Onset and duration of immunity and minimum dosage with CU cholera vaccine in turkeys via drinking water

Growing turkeys were partly protected against fowl cholera 4 days after vaccination with the live Clemson University (CU) strain of Pasteurella multocida administered in drinking water, and they were highly protected from 1 to 4 weeks after vaccination. The commercially available lyophilized vaccine and the freshly cultured vaccine of the CU strain did not differ in the level of immunity induced. Immunity was relatively high in turkeys vaccinated with 1:2 and 1:4 dilutions of the recommended dosage (4 X 10(8) P. multocida) but was significantly (P less than 0.05) lower in turkeys vaccinated with a 1:8 dilution of the recommended dosage. Immunity continued for 13 weeks after the last vaccination in turkeys vaccinated twice 3 weeks apart, but it persisted for only 8 weeks in those vaccinated only once.     

Comparison of live avirulent M-9, Minnesota, and CU fowl cholera vaccines

The M-9 and Minnesota (MN) avirulent Pasteurella multocida vaccines were evaluated and compared with the Clemson University (CU) vaccine, which had been shown to be highly effective in preventing fowl cholera in turkeys. Neither the M-9 nor the MN vaccine given in the drinking water was as effective as the CU vaccine in protecting turkeys against challenge with virulent P. multocida. When grown in brain-heart infusion (BHI) agar as recommended, the M-9 was not as efficacious as when it was grown in BHI broth. The M-9 was as effective as the CU vaccine only when grown in BHI broth and given at 10 times the standard dosage. Injection of the M-9 vaccine into the air spaces of the head at a site near the caudal rim of the ear after one vaccination in the drinking water was not as effective for hyperimmunizing potential breeders as was the CU vaccine injected at the same site. A microtiter agglutination test demonstrated a significant (P less than 0.05) correlation between the level of anti-P. multocida antibody found 1 week after vaccination and survival after challenge with virulent P. multocida.     

Immune response of poults following intranasal inoculation with Artvax vaccine and a formalin-inactivated Bordetella avium bacterin

Poults 3 weeks and older developed temporary tracheal resistance to intranasal challenge following inoculation of either Artvax vaccine or formalin-inactivated Bordetella avium bacterin by the intranasal and eyedrop routes. Resistance usually persisted for 3-4 weeks after B. avium challenge. However, with constant exposure to infected controls, the vaccinated birds eventually developed tracheal infection. Day-old poults did not respond to either the Artvax or the bacterin and were completely susceptible to challenge. Two-week-old poults responded to some degree, but poults 3 weeks old and older responded best. Poults inoculated with bacterin by aerosol or by drinking water did not respond as well as those inoculated by the intranasal and eyedrop routes. When poults were given a single subcutaneous injection at 3 weeks of age and challenged 2 weeks later, three of five resisted infection for 18 days.     

Virulence and toxigenicity of capsular serogroup D Pasteurella multocida strains isolated from avian hosts

Five capsular serogroup D strains of Pasteurella multocida isolated from avian hosts were examined for virulence and toxigenicity. Virulence was based on development of lethal infections or lesions following intramuscular exposure of turkey poults. The four strains isolated from turkeys varied from slightly to moderately virulent; the strain isolated from a chicken was avirulent. Poults exposed by intra-airsac inoculation with relatively few organisms of the more virulent of the strains had a high mortality rate; however, intranasal exposure of poults with this strain did not cause clinical disease or establish infections. All strains from turkeys were toxigenic, producing heat-labile toxins that killed poults when administered intraperitoneally and caused focal dermal lesions when administered intradermally. Using these criteria, the strain from a chicken was not toxigenic. The demonstration of virulence, particularly the high mortality in poults exposed via air sacs, indicates avian capsular serogroup D strains are a potential cause of fowl cholera.     

Vaccination of turkeys with cell-free culture filtrate of Pasteurella multocida

Turkeys given cell-free culture filtrate (CCF) of Pasteurella multocida strain R44/6 orally, via air sacs, or subcutaneously mixed 1:1 with incomplete Freund's adjuvant (IFA) at 6 and 9.5 weeks of age were compared with negative controls given bacteriologic medium and positive controls vaccinated with a commercial bacterin. At 13 weeks of age, serum antibody titers to P. multocida were detectable only in turkeys given CCF in IFA (low titers) and positive control turkeys (high titers), at which time turkeys were challenged orally with either the homologous strain or strain P-1059. Protection against challenge with strain R44/6 was provided by the commercial bacterin, CCF in IFA, and CCF given via air sacs. When turkeys were challenged with strain P-1059, protection was superior in turkeys given CCF via air sacs, intermediate in turkeys given commercial bacterin or CCF in IFA, and absent in negative control turkeys and turkeys given CCF orally. These results indicate CCF is an effective immunogen when administered via the lower respiratory tract for protecting turkeys against pasteurellosis.     

Attenuated live fowl cholera vaccine. I. Development of vaccine strain M3G of Pasteurella multocida

A live cholera vaccine was developed from a virulent avian septicemia strain of Pasteurella multocida serotype 1. The virulent parental strain was mutagenized with N-methyl-N'-nitro-N-nitroso guanidine. Mutants were selected that had either smaller colonies at 37 C or temperature sensitivity for growth at 41 C. Four small-colony mutants and 2 temperature-sensitive mutants were studied. All the mutants were avirulent for turkeys. Sixteen days after turkeys were vaccinated with each mutant, both the vaccinates and unvaccinated controls were challenge-exposed to virulent P. multocida of the homologous serotype and the heterologous serotype 3. Two of the small-colony mutant strains protected against both homologous and heterologous challenge. Suggested for a live cholera vaccine is P. multocida M3G, a small-colony-forming mutant, innocuous for both mice and turkeys and stable against reversion.     

Evaluation of combined Pasteurella vaccines in control of sheep pneumonia

The effectiveness of an oil adjuvant vaccine (OAV) incorporating locally isolated strains of Pasteurella haemolytica type 7 and Pasteurella multocida types A and D was compared with that of Carovax (Wellcome Laboratories) in imported cross-bred lambs. The criterion of efficacy was the ability of the vaccines to reduce the extent of pneumonic lesions in vaccinated as against unvaccinated control lambs. The OAV produced at this Institute significantly reduced the lung lesions at P less than 0.05 level compared with its control group when challenged with P. haemolytica alone. However, the vaccine was unsatisfactory against P. multocida or combined P. multocida P. haemolytica challenge. Carovax did not produce any significant reduction in the lung lesions caused by P. haemolytica and/or P. multocida.     

Septicemia in vaccinated and nonvaccinated turkeys inoculated with Pasteurella multocida serotype A:3,4

Sixty-four, 10-week-old turkeys were inoculated with a highly virulent field isolate (86-1913) of Pasteurella multocida serotype A:3,4 by an oculo-nasal-oral route. Inoculated turkeys were examined at 4, 8, 16, 20, and 24 hours post-inoculation for bacteremia and histologic lesions. Bacteremia was detected in one of six turkeys 8 hours after inoculation and in four of six turkey poults at 16 hours post-inoculation. Pasteurella multocida was isolated from the spleens of two turkeys at 8 hours and from the spleens of all six poults 16 hours after inoculation. Peak concentrations of P. multocida reached 10(9) colony forming units per ml of blood. At 4 to 8 hours post-inoculation, isolate 86-1913 produced a fibrinopurulent bronchopneumonia followed by severe pulmonary necrosis, pleuritis, vasculitis; and, at 16 to 24 hours post-inoculation numerous extracellular bacteria were observed. Hepatic lesions included focal heterophil aggregates 8 hours after inoculation; these progressed to hepatic necrosis. Numerous extracellular bacteria within sinusoids were present 16 to 24 hours after inoculation. At 16 to 24 hours post-inoculation, there was degeneration of periarteriolar reticular cells in the spleen; these cells progressed to coalescing coagulative splenic necrosis with extracellular bacterial colonies. A second group of 41, 10-week-old turkeys, previously vaccinated with the Clemson University strain of P. multocida serotype A:3,4, were challenged with isolate 86-1913.(ABSTRACT TRUNCATED AT 250 WORDS)     

Efficacy and safety of a cell-culture live virus vaccine for hemorrhagic enteritis of turkeys: laboratory studies

Poults free from hemorrhagic enteritis (HE) antibody were vaccinated by gavage at 1 day or 2 weeks of age with a live HE vaccine virus that had been propagated in a Marek's disease (MD)-induced B-lymphoblastoid cell line of turkey origin. Vaccinated and unvaccinated poults were challenged with a virulent HE virus at various times postvaccination. One hundred tissue-culture-infectious doses of the vaccine virus per poult were sufficient to induce a serological response as well as to protect poults against HE lesions and mortality. Vaccinated poults were protected against the disease as early as 1 week and as late as 8 weeks PV. The vaccine was efficacious by several routes of application. The vaccine virus spread horizontally from vaccinated to contact-exposed poults, as indicated by seroconversion and resistance of contact-exposed poults to challenge. The vaccine had no detectable harmful effects on the humoral immune response to particulate antigens or on weight gain of vaccinated poults. The vaccine proved to be free from MD virus, as indicated by the absence of MD lesions and antibody in 8-week-old chickens inoculated intra-abdominally with the vaccine at hatching. These findings indicate that the cell-culture-propagated HE vaccine is efficacious and safe.     

Protection of piglets against atrophic rhinitis by vaccinating the sow with a vaccine against Pasteurella multocida and Bordetella bronchiseptica

The efficacy of a new vaccine against atrophic rhinitis in pigs was tested in the Netherlands and Denmark. The vaccine contained protein dO (a truncated Pasteurella multocida toxin which is immunogenic and non-toxic), inactivated Bordetella bronchiseptica whole cells, and an adjuvant. The sows were either vaccinated intramuscularly with 2 ml of the vaccine at six to eight and two to four weeks before expected farrowing or left unvaccinated as controls. All the piglets were challenged intranasally with B bronchiseptica when three to seven days old and with P multocida three to four days later. Pigs born to the vaccinated sows performed significantly better than pigs born to the control sows when judged on growth, average daily weight gain and snout scores. The challenge organisms were reisolated more frequently from the control pigs than from the pigs in the vaccinated group. The vaccinated sows and their progeny developed high titres of antibodies against B bronchiseptica and P multocida toxin.     

Lysates of turkey-grown Pasteurella multocida: effects of solubilizing agents on the immunologic properties of membrane vesicles

Membrane vesicles from lysed suspensions of turkey-grown Pasteurella multocida were treated with various solubilizing agents to release protein that may contain cross-protection factor. Potassium thiocyanate, NaOH-glycine, lithium diiodosalicylate, guanidine hydrochloride, n-butanol, dimethyl sulfoxide, Triton X-100, and sodium lauryl sarcosinate were each tested as solubilizing agents. Vaccines made from combining solubilized membrane vesicles with complete lysate supernatant fluid produced various degrees of protection against challenge exposure with a heterologous serotype of P multocida in turkeys. Only vaccines prepared from membranes that were solubilized with potassium thiocyanate and sodium lauryl sarcosinate protected as well as complete lysate from turkey-grown P multocida. The amount of protein in each vaccine did not relate to protection. Distinct chemical differences were observed between lysates prepared from turkey-grown P multocida and lysates prepared from 41 C broth-grown P multocida. The external morphology of P multocida, after treatment with lysozyme and EDTA, was similar whether grown in broth or in turkeys.     

Fowl cholera immunity induced by various vaccines in broiler minibreeder chickens determined by enzyme-linked immunosorbent assay

Broiler minibreeder hens were vaccinated for protection against fowl cholera at 12 and 21 weeks of age using several vaccination schemes, which included a live Pasteurella multocida (CU strain) vaccine, two commercial polyvalent fowl cholera oil-based bacterins, and two experimentally prepared polyvalent oil-based bacterins. Some treatment groups received only live or killed vaccines, whereas others received a live vaccine at 12 weeks followed by a killed product at 21 weeks. At 42 weeks of age, all birds that received the live CU vaccine twice or once followed by a bacterin survived challenge. Birds that received killed vaccines only were significantly less protected but still showed a respectable survival rate of 86%. All unvaccinated controls died within 72 hr after challenge. At 72 weeks of age, overall protection was lower than that at 42 weeks, regardless of vaccination treatment. Antibody titers were usually higher in birds that received bacterins than in those receiving live vaccines, yet overall protection was still greater in those birds that received the live cholera vaccine twice.