Effects of adjuvant-augmented germling vaccines in turkey poults challenged with Aspergillus fumigatus

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.     

Protection of turkey poults from Bordetella avium infection and disease by pili and bacterins

The role of pili in protection against Bordetella avium infection in turkey poults was studied. An isolate that produced the largest number of pili under growth conditions developed in our laboratory was used for preparation of pili and bacterin and for challenge. The pili were isolated, purified, examined by electron microscopy, and tested for purity by gel electrophoresis. Poults were vaccinated with oil-adjuvant pili, formaldehyde- or merthiolate-inactivated bacterins, or a commercial bacterin. Poults were vaccinated once or twice subcutaneously at different ages and challenged intranasally with a pathogenic B. avium isolate 5 days following the last vaccination. A few vaccinated birds had very mild clinical signs. B. avium was isolated from the sinuses of a few vaccinated birds, and growth was scanty. The mean colony counts from tracheal sections was significantly higher (P less than 0.1) in unvaccinated challenged poults than in vaccinated challenged poults. It is postulated that B. avium pili are important immunogens in turkey poults.     

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.     

Use of an enzyme-linked immunosorbent assay to measure antibody levels in turkey breeder hens, eggs, and progeny following natural infection or immunization with a commercial Bordetella avium bacterin

Twelve large white turkey hens were immunized with a commercially available Bordetella avium bacterin. Hens and eggs were tested using an enzyme-linked immunosorbent assay (ELISA) to determine the response to the bacterin. Three hundred poults were then obtained from two commercial flocks, the hens of one flock having been immunized with the same bacterin used on the group of 12 turkeys. Titers of the poults were monitored for 7 weeks, and poults were challenged by exposure to infected poults at 1, 7, 14, and 21 days post-hatch. Hens produced an antibody response following immunization, with a parallel antibody response being detected in eggs. Maternal antibodies were present in poults from immunized hens. Poult titers declined to near the level of poults from unimmunized hens by 14 days of age. Poults from immunized hens challenged at 1 and 7 days were resistant to development of clinical disease and gross lesions, whereas all poults from unimmunized hens exhibited clinical signs and gross lesions. After 14 days, the resistance of both groups to development of clinical disease, became near equal, neither group being affected as severely as the unimmunized hens challenged at days 1 and 7. Six commercial turkey breeding flocks and their progeny that had not been vaccinated for B. avium and had no history of B. avium infection were evaluated with the B. avium ELISA. There were variations between the flocks, with poult titers reflecting those found in the hens.     

Efficacy of viable and inactivated Newcastle disease virus vaccines in turkeys

Market turkeys spray-vaccinated at 20 days of age with viable Newcastle disease virus (NDV) vaccine and challenged 7 weeks postvaccination failed to yield NDV by tracheal swabbing 4 days postchallenge but demonstrated serologic evidence of infection. Birds vaccinated subcutaneously with inactivated oil-emulsion (OE) NDV vaccine had virologic and serologic evidence of infection. Breeder hens vaccinated by spray with commercial La Sota vaccine at 19 weeks of age and revaccinated subcutaneously with OE vaccine at 32 weeks of age had an adequate level of resistance against a drop in egg production but demonstrated serologic evidence of infection when challenged with velogenic NDV at 38 weeks of age.     

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.     

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.     

Propagation of virulent and avirulent turkey hemorrhagic enteritis virus in cell culture

Virulent and apathogenic isolates of turkey hemorrhagic enteritis virus (HEV) were successfully propagated in lymphoblastoid cell lines of turkey origin, whereas spleen and kidney cell cultures from HEV-infected turkeys failed to replicate the virus. The lymphoblastoid cell lines used were MDTC-RP16 and MDTC-RP19, which were previously established from tumors induced by Marek's disease virus in turkeys. Virus replication followed co-cultivation of lymphoblastoid cells with spleen cells from HEV-infected turkeys. Virus replication was demonstrated by immunofluorescence, by agar-gel-precipitin tests, and by electron microscopy. Supernatant fluid of cultures infected with virulent HEV caused death and specific lesions in turkey poults. Poults vaccinated with apathogenic HEV were protected against death and lesions after challenge with pathogenic HEV, which was recovered from infected cultures. The MDTC-RP19 cell line appeared far more susceptible than the MDTC-RP16 cell line to infection with HEV.     

Passively administered antibodies alleviate stunting syndrome in turkey poults

Stunting syndrome is an enteric disease of young turkeys that results in reduced growth (stunting) of poults and impaired feed efficiency. A virus, which has been termed the stunting syndrome agent (SSA), causes stunting syndrome. In this study passive immunity was evaluated as a method of protecting poults from stunting syndrome. One-day-old poults were injected with either tryptose phosphate broth, an anti-SSA antibody preparation, or an anti-Newcastle disease virus antibody preparation before challenge by placing them into SSA-contaminated isolators or control (nonchallenge) isolators. Poults that received anti-SSA antibodies were significantly heavier (P < 0.05) and did not display as severe clinical disease compared to birds that did not receive the anti-SSA antibodies. However, the birds that received anti-SSA antibodies and were challenged were significantly lighter (P < 0.05) than birds that were not challenged. The results of this trial demonstrate that the injection of anti-SSA antibodies benefited poults undergoing stunting syndrome. The role of passive immunity, either through breeder hen vaccination or through supplying antibodies to poults artificially (i.e., at the hatchery), may have future applications in alleviating stunting syndrome.     

Observations on the pathogenicity of Alcaligenes faecalis in chickens

A series of trials was conducted in which specific-pathogen-free (SPF) leghorn chicks were exposed to various isolates of Alcaligenes faecalis. Chicks were inoculated with A. faecalis alone or in combination with Newcastle disease/infectious bronchitis (Nc/Br) vaccine, laryngotracheitis vaccine, infectious bursal disease virus, or Mycoplasma gallisepticum. The response was evaluated by morbidity, mortality, airsacculitis, reisolation of A. faecalis, and histopathological lesions of tracheas. Although A. faecalis was recovered up to 42 days postinoculation in some cases, no clinical signs were directly attributed to simple A. faecalis infection. None of the other agents significantly increased the severity of A. faecalis signs or lesions, except that A. faecalis-infected chicks that were given Nc/Br vaccine had prolonged microscopic tracheal lesions. In another trial, the effects of A. faecalis in young SPF leghorns, non-SPF broilers, and turkeys were compared. Broiler-type chicks were more susceptible than leghorns and less susceptible than poults. Consequently, the use of leghorns as a model for studying this infection is questioned.     

Minimization of pathologic changes in Ornithobacterium rhinotracheale infection in turkeys by temperature-sensitive mutant strain

The protection elicited by a temperature-sensitive (Ts) mutant of Ornithobacterium rhinotracheale (ORT) vaccine against challenge with pathogenic strain was investigated. In Experiment 1, specific serologic response to ORT was detected in 12%-19% of Ts-vaccinated birds at 3 wk postvaccination by either drinking water or oculo-nasal instillation. At 7 days postchallenge, 100% of Ts-vaccinated turkeys of all groups were able to respond with an ORT-specific antibody response, but the control group was not, suggesting the potential of Ts strain to evoke immune protection. The study also revealed a statistically significant ability of the Ts strain to protect vaccinated turkeys against gross lesions caused by the pathogenic strain of ORT in treated groups vs. control. In Experiment 2, seroconversion was detected by enzyme-linked immunosorbent assay in birds after they were given the Ts strain in drinking water in field conditions. The results of the field study showed mean scores of gross lesions of nonvaccinated/challenged groups to be up to seven times higher than those of the vaccinated/challenged group. In addition, reisolation rates and quantification of ORT colonies per gram of lung tissue were significantly lower for vaccinated/challenged than for nonvaccinated/challenged turkeys. In conclusion, results from laboratory and field experiments suggest that use of the Ts mutant strain of ORT as a live vaccine would be a suitable method to evoke protection against ORT infection in turkeys.     

Lack of protection against Bordetella avium in turkey poults exposed to B. avium-like bacteria

Six laboratory experiments were designed to determine whether poults infected with the nonpathogenic Bordetella avium-like (BAL) bacteria would develop immunity to B. avium (BA), the causative agent of turkey coryza. The BAL bacteria were isolated from poults given that organism, but few colonies were observed by 3 weeks postexposure. No serum-agglutinating antibody to the BAL bacterium was detected in poults exposed to that organism. Poults exposed to BAL bacteria either once or twice at different ages were not protected from infection or disease following experimental challenge between 1 and 7 weeks of age with pathogenic BA.     

An enzyme-linked immunosorbent assay for detection of Bordetella avium infection in turkey flocks: sensitivity, specificity, and reproducibility.

An enzyme-linked immunosorbent assay (ELISA) for detection of Bordetella avium infection in turkey poults was developed. One-week-old poults challenged intratracheally with 10(12) colony-forming units of B. avium had detectable titers (greater than or equal to 11), with an average of 13.6% positive samples when the birds were 6 to 11 weeks old. The method was sensitive enough to detect maternal antibodies to B. avium in poults up to 3 weeks of age. The same poults challenged at 1 week of age had 100% tracheal infection up to 3 weeks of age, which dropped to 0% by 6 weeks. The method resulted in no false-positive samples (titer = 0) from birds not infected with B. avium and tested weekly between 4 and 11 weeks of age. Antibodies in turkey flocks infected with Newcastle disease virus, hemorrhagic enteritis virus, and Mycoplasma meleagridis, and birds infected with Escherichia coli had no apparent cross-reactivity to the B. avium antigens used in the ELISA. The percentages of B. avium-positive serum samples collected from different turkey flocks did not significantly differ (P greater than 0.05) when samples were tested by the developed ELISA at different times, an indication of the reproducibility of the method.     

Protection of laying hens against infectious bronchitis with inactivated emulsion vaccines

Commercially-reared laying chickens were challenged at 31 weeks of age with a virulent infectious bronchitis (IB) virus. They showed a sharp drop in egg production, despite having been vaccinated at four and eight weeks old with live attenuated IB vaccines to a recommended schedule. In contrast, similar birds that had been further immunised at point-of-lay with inactivated oil emulsion IB vaccine, or with a combined IB/Newcastle disease (ND) emulsion vaccine, showed no detectable fall in egg production after the same challenge. Unvaccinated susceptible specific pathogen-free birds challenged at the same time stopped laying almost completely. In the birds revaccinated with emulsion vaccine, measurement of haemagglutination inhibition antibody levels to IB showed their geometric mean titres to be raised from less than 5 log2 at the time of vaccination to over 10 log2 four weeks later. Their antibody levels did not rise further followining the IB challenge whereas in the birds that had not been revaccinated antibody rises to nearly 10 log2 were detected after the same challenge. For pullets vaccinated earlier with live IB vaccine, revaccination with inactivated IB or IB/ND oil emulsion vaccine at point-of-lay provides a safe and effective way of protecting their egg production against IB infection.     

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.     

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.     

The prevalence of Chlamydia psittaci infections in Belgian commercial turkey poults

The prevalence of Chlamydia psittaci infections in Belgian commericial turkey poults was examined and a follow-up study of one Belgian turkey flock was performed. Sera were examined for the presence of anti-chlamydia antibodies by immunoblotting. Cloacal and conjunctival swab smears and lung impression smears were examined for the presence of chlamydial antigen using the IMAGEN Chlamydia immunofluorescence test. Anti-chlamydia antibodies were found in 90 of 100 sera collected at slaughter from turkeys raised during the summer of 1992. The following winter, 73 of 100 sera reacted positively. On all twenty farms examined during 1992, turkeys were positive for anti-chlamydial antibodies. During 1993, chlamydial antigen was detected in swabs from 20 of 40 slaughterhouse turkeys tested. Antigen was found more often in the cloaca than in the conjunctiva. Chlamydial antigen was detected in samples from each of the 4 farms examined. The follow-up study on a turkey farm, sampling the birds at weekly intervals from one week old until 12 weeks of age, revealed that chlamydial antigen and anti-chlamydial antibodies were present during the whole period. During 1994, chlamydial antigen was detected in 45 of 60 lungs from slaughterhouse turkeys from all of 6 farms. During 1995, chlamydial antigen was detected in 41 of 54 lungs of 6 week old commercial turkey poults. The results of the present study indicate that Chlamydia psittaci infections are highly prevalent amongst Belgian commercial turkey poults with apparently little seasonal or year-to-year variation and that turkeys can contract the infection at an early age.     

The efficacy of a temperature-sensitive mutant vaccine against Northwest Arkansas isolates of Alcaligenes faecalis

The efficacy of a commercially produced temperature-sensitive mutant Alcaligenes faecalis vaccine was evaluated in turkeys contact-challenged with one of three strains of A. faecalis. In the vaccinated control group, the vaccine strain of A. faecalis colonized the nasal turbinates but not the trachea, caused no clinical signs of turkey coryza, and induced humoral antibodies. In the vaccinated challenged groups, the vaccine reduced both the severity of lesions and the number of birds exhibiting lesions compared with unvaccinated challenged groups, but it did not prevent colonization of challenge strains of A. faecalis.     

La Sota vaccination may not protect against virus shedding and the lesions of velogenic Newcastle disease in commercial turkeys

The aim of this project is to study the clinical signs and lesion of velogenic Newcastle disease (vND) in commercial turkeys, and also to find out if La Sota vaccination offered protection against these signs and lesions. The cockerels were included as positive controls. One hundred and twenty turkey poults and cockerels were divided into eight groups as follows: unvaccinated unchallenged turkeys (UUT), unvaccinated challenged turkeys (UCT), vaccinated unchallenged turkeys (VUT), vaccinated challenged turkeys (VCT), and along the same lines, the cockerel groups were UUC, UCC, VUC and vaccinated challenged cockerels (VCC). Vaccination was at 3 weeks of age while challenge was at 6 weeks of age. The unvaccinated turkeys and cockerels (UCT and UCC) showed different degrees of depression, diarrhoea and later paralysis at challenge. Total mortality was 100% in cockerels within 6 days, but 60% in turkeys. Similar but milder clinical signs were found in the VCC with a total mortality of 13.3%. The VCT showed mild drop in feed and water consumption, and no mortality. All the challenged groups had significant (p < 0.05) loss of weight when compared with their controls. Necropsy showed that while the UCC had severe proventricular haemorrhages, intestinal and caecal tonsil ulcers, the UCT had no digestive tract lesion. There was severe atrophy of the lymphoid organs in all the challenged groups. Histopathological sections of the bursa, spleen and thymus in all the challenged groups with special emphasis on the vaccinated and unvaccinated turkeys with mortalities of 0 and 60%, respectively, had very severe necrosis and depletion of the lymphoid tissue. Virus was isolated from the cloacal swabs. The haemagglutination inhibition antibodies were significantly higher (p < 0.05) in the challenged groups than the unchallenged. The above observations in the intestinal tracts of UCT are of diagnostic significance while the gross and microscopic lesions in the UCT and VCT show that La Sota vaccination may not protect turkeys against the destruction of the lymphoid organs by vND as earlier reported in chickens. This may lead to immunosuppression and production problems in areas where vND is enzootic.     

Marek's disease in turkeys: lack of protection by vaccination

Herpesvirus of turkeys, a highly effective vaccine against Marek's disease (MD) in chickens, was ineffective in protecting turkeys against MD. Another tissue-culture attenuated vaccine virus also protected chickens, but not turkeys, from MD. Intact and immunosuppressed turkey poults inoculated with herpesvirus of turkey developed a persistent viremia, but did not have detectable gross or microscopic lesions.