Safety and efficacy of the Mycoplasma synoviae MS-H vaccine in turkeys

The live, attenuated, temperature-sensitive Mycoplasma synoviae (MS) vaccine strain MS-H is used to control virulent MS infection in commercial chicken flocks. However, the safety of this vaccine and its potential to prevent disease in turkeys have not been investigated. In this study, MS-H was shown to colonize the upper respiratory system and to induce an antibody response in turkeys but, even at the maximum release dose, was not found to cause air sac, joint, or tracheal lesions typical of wild-type MS infection. Histopathologic examinations of the vaccinated turkeys after exposure to a virulent MS challenge revealed that administration of the vaccine by aerosol, but not eye drop, at the dose recommended for chickens protected the birds against microscopic lesions and colonization of the virulent MS in trachea. It is concluded that MS-H vaccine is safe for use in turkeys and, when used as aerosol at the dose recommended for commercial chickens, can protect turkeys against tracheal lesions caused by a wild-type MS strain.     

Vaccination of turkeys with an avian pneumovirus isolate from the United States

Four-week-old poults obtained from avian pneumovirus (APV) antibody-free parents were vaccinated with different serial 10-fold dilutions of cell culture-propagated APV vaccine. The birds were vaccinated with 50 microl into each conjunctival space and nostril (total of 200 microl). Each poult of each group was vaccinated in groups that received doses of 4 x 10(4), 4 x 10(3), 4 x 10(2), 4 x 10(1), or 4 x 10(0) 50% tissue culture infective dose (TCID50) of APV vaccine, respectively. Respiratory signs were seen between 3 and 12 days postvaccination (PV) in the poults that were vaccinated with 4 x 10(4), 4 x 10(3), and 4 x 10(2) TCID50, respectively. In these groups, APV was detected from swabs collected at 5 days PV and seroconversion was detected at 2 wk PV. The groups that were originally vaccinated with 4 x 10(1) and 4 x 10(0) TCID50 developed mild clinical signs after vaccination, but neither virus nor antibody was detected PV. At 2 wk PV (6 wk of age), birds from each group, along with five unvaccinated controls, were challenged with APV. Upon challenge, the 4 x 10(4) and 4 x 10(3) TCID50 groups were protected against development of clinical signs and were resistant to reinfection. The group previously vaccinated with 4 x 10(2) TCID50 developed clinical signs after challenge that were considerably milder than those seen in the groups that had previously been vaccinated with lower doses or no virus. Even though 4 x 10(2) TCID50 vaccine dose administered by intranasal ocular route resulted in infection, incomplete protection resulted with this pivotal dose. Upon challenge, the 4 x 10(1) and 4 x 10(0) TCID50 groups exhibited milder disease signs than those seen in the challenged unvaccinated controls. In these groups, APV was detected in preparations of swabs collected at 5 days postchallenge (PC) and seroconversion was detected at 2 wk PC. These results indicate that the dose of APV vaccine that causes protection is higher than that required to produce infection.     

Comparison of different attenuation strategies in development of a Salmonella hadar vaccine

The purpose of this work was to develop a live, attenuated vaccine strain to protect chickens against colonization by group C Salmonella. We constructed two candidate vaccines: a deltacya deltacrp derivative and a deltaphoP derivative of Salmonella hadar. White Leghorn chickens were vaccinated at day of age and at 2 wk with one of the two strains. A nonvaccinated group served as a control. At 4 wk of age, all birds were challenged with wild-type S. hadar and necropsied 6 days later. Numbers of S. hadar in the ceca were determined. Enzyme-linked immunosorbent assay-derived serum immunoglobulin G responses against S. hadar lipopolysaccharide indicated that both strains induced a serum antibody response. The average optical density450 for birds vaccinated with the deltaphoP or deltacya deltacrp derivatives was 0.456 and 0.881, respectively. Although the deltacya deltacrp derivative induced higher levels of serum antibody, it did not provide an immune response protective against colonization by S. hadar. Conversely, birds vaccinated with the deltaphoP strain showed significant protection against S. hadar challenge. Seventy percent of the nonvaccinates, 60% of the deltacya deltacrp vaccinates, and 15% of deltaphoP vaccinates were positive for S. hadar in tissues. In a second experiment, birds were vaccinated with either the deltaphoP strain or buffer and challenged with a 10-fold higher dose than in the first experiment. After challenge, all of the birds in both groups were colonized. The geometric mean number of cecal S. hadar isolated from the control group was 1.0 x 10(6) colony-forming units (CFU)/g, and from the vaccinated group, this value was 32 CFU/g, indicating a four to five log reduction in colonization by the challenge strain.     

Evaluation of the non-temperature-sensitive field clonal isolates of the Mycoplasma synoviae vaccine strain MS-H

The live attenuated temperature-sensitive (ts+) Mycoplasma synovia (MS) strain, MS-H, is used as a vaccine in a number of countries to control virulent MS infection in commercial chicken flocks. Nine out of 50 isolates made from flocks vaccinated with MS-H were found to have lost the ts+ phenotype of the original vaccine strain. In order to examine the influence of the ts- phenotype on virulence of the isolates, four of the ts- isolates, the MS-H vaccine, and the vaccine parent strain 86079/7NS were administered by aerosol in conjunction with infectious bronchitis virus to 3-wk-old specific-pathogen-free chickens. The four ts- clones induced only minimal air sac lesions that were not different in severity from those caused by MS-H vaccine; however, the vaccine parent strain 86079/7NS caused air sac lesions that were significantly greater than those of MS-H and all ts- clones. The vaccine parent strain 86079/7NS and two of the ts- clones were recovered from the air sacs of the respectively infected chickens whereas the MS-H vaccine and two other ts- clones were not. Three of the ts- isolates caused increased tracheal mucosal thicknesses that were significantly greater than those from birds inoculated with MS-H, and one caused increased tracheal mucosal thicknesses that were significantly less than those from birds inoculated with 86079/7NS. In conclusion, unlike the MS-H vaccine, the MS-H ts- clones were associated with minor changes in tracheal mucosa; however, unlike the vaccine parent strain, they did not induce lesions in the air sacs. These results suggest that factors other than ts+ phenotype are involved in the attenuation of the MS-H vaccine.     

Duration of immunity with Mycoplasma synoviae: comparison of the live attenuated vaccine MS-H (Vaxsafe MS) with its wild-type parent strain, 86079/7NS

The duration of protective immunity elicited by the MS-H vaccine was evaluated by experimental challenge of chickens at 15 and 40 wk after eyedrop vaccination. Immunity induced by the parent strain of the vaccine, 86079/7NS, was also investigated for comparison. A serological response to Mycoplasma synoviae was detected in 89% to 100% of MS-H vaccinates and 86079/7NS inoculates at 15, 27, 30, 35, and 40 wk after inoculation. A significantly lower incidence of air-sac lesions and lower air-sac lesion severity were observed in both the MS-H vaccinated and the 86079/7NS inoculated groups, as compared to the unvaccinated controls, after both challenge points. Tracheal mucosal thicknesses in MS-H vaccinates was significantly lower in the upper, lower, and total trachea at 40 wk after vaccination, as compared to the controls. It was demonstrated in this experiment that protective immunity, as determined by protection against experimental challenge, was maintained to at least 40 wk after vaccination.     

Onset of immunity with Mycoplasma synoviae: comparison of the live attenuated vaccine MS-H (Vaxsafe MS) with its wild-type parent strain (86079/7NS)

The onset of protective immunity with MS-H was determined through experimental challenge and compared with the parent strain 86079/7NS. MS-H vaccinates and 86079/7NS inoculates were challenged at 1, 2, 3, 4, 5, and 6 wk after vaccination, then examined 2 wk after challenge for signs of respiratory disease. Serologic results indicated that 100% of MS-H vaccinates had antibodies to MS by 3 wk after vaccination and 100% of 86079/7NS inoculates were positive by 2 wk after inoculation. From 3 wk after vaccination, MS-H vaccinates had a significantly lower incidence of air sac lesions and, from 4 wk after vaccination, a significantly lower air sac lesion severity. In 86079/7NS-inoculated birds, a significantly lower incidence of air sac lesions was observed from 1 wk after inoculation, and air sac lesion severity was significantly lower than the unvaccinated controls at 3 wk after inoculation. It would appear that, under the conditions of this experiment, protective immunity elicited by MS-H appeared at 4 wk after vaccination, slightly later than the appearance of serum antibody. Although the MS-H vaccine was slower to establish protective immunity than 86079/7NS, there was no significant difference between the two strains by 4 wk after vaccination or inoculation.     

Validation of a disease model in Japanese quail (Coturnix coturnix japonica) with the use of Escherichia coli serogroup O2 isolated from a turkey

This study established a disease model and protocol for bacterial challenge with Escherichia coli serogroup O2 strain EC317 in Japanese quail. Five groups of 10 birds each were injected subcutaneously in the breast with 200 μL of a brain-heart infusion (BHI) culture containing 1 × 10(8), 1 × 10(7), 1 × 10(6), 1 × 10(5), or 1 × 10(4) colony-forming units/mL of the test organism, which had been isolated from a turkey with cellulitis and septicemia. Birds in a 6th group were controls that received sterile BHI alone. Localized lesions of cellulitis developed in all of the birds that received E. coli. The morbidity and mortality rates were highest (100%) in the birds receiving the highest dose of E. coli and decreased linearly with decreasing dose (P < 0.05). Severity of disease, including lesions of pericarditis and perihepatitis, was also directly proportional to the dose of E. coli. These findings indicate that this disease challenge protocol can be used to study disease resistance and immunologic consequences of contaminant exposure or other stressors in birds.     

A comparison of the onset of protection induced by Newcastle disease virus strain B1 and a fowl poxvirus recombinant Newcastle disease vaccine to a viscerotropic velogenic Newcastle disease virus challenge

Four-week-old specific-pathogen-free white rock chickens were immunized with either a commercial recombinant fowl poxvirus-vectored Newcastle disease vaccine (FPN) expressing the hemagglutinin-neuraminidase and fusion protein genes of Newcastle disease virus (NDV) strain B1 or live NDV B1. Vaccinates and controls were challenged by eyedrop and intranasal (E/I) route with a viscerotropic velogenic NDV at 14 days postvaccination to determine the time of clearance of challenge virus. In a subsequent experiment, chickens were challenged at 3, 6, or 10 days postvaccination to determine the onset of immunity. Chickens that received a recommended field dose (1x) or a 0.01x dose of FP-N subcutaneously (s.c.) and were seropositive by hemagglutination-inhibition test at 14 days postvaccination cleared the challenge virus by 14 days postchallenge. Clinical Newcastle disease and high challenge virus titers in tissues were seen only in seronegative FP-N 0.01x dose vaccinates and controls. In a comparison of vaccination with FP-N (1x, 10(4,9) median tissue culture infective dose) s.c., B1 (10(6) median egg infective dose [EID50]) s.c., or B1 (10(6) EID50) E/I, chickens vaccinated at 6 or 10 days before challenge with all vaccines were protected against clinical disease, but only those vaccinated with B1 E/I 10 days before challenge were protected against infection with the challenge virus. Vaccination at 3 days before challenge with B1 E/I provided early protection, but severe nervous signs developed later and reduced overall protection to 60%, whereas disease in chickens vaccinated with B1 s.c. and FP-N s.c. 3 days before challenge was similar to the challenge controls.     

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.     

Elevation of muscle and plasma 3-methylhistidine as a result of turkey coccidiosis

To assess muscle breakdown during avian coccidiosis, the level of the non-metabolizable amino acid 3-methylhistidine (3MH) was determined in muscle and plasma from turkey poults that received an infection with a field isolate containing a mixture of Eimeria species. The effect of increased levels of parasitism was evaluated at 6 days postinoculation (DPI) in birds receiving 2.5 x 10(4), 1 x 10(5), or 2 x 10(5) oocysts each. The changes in 3MH levels during recovery from acute infection were assessed at 6-29 DPI in animals given 1.9 x 10(5) oocysts per bird. In some experiments, uninoculated birds given the same amount of feed as infected birds (pair fed) were used to determine the impact of feed deprivation on weight loss and 3MH levels. Infected birds had significantly elevated plasma and muscle 3MH at 6 DPI after a single dose of Eimeria oocysts. The plasma and muscle 3MH returned to control levels after 14 DPI. The 3MH levels increased with increased dose of oocysts. Plasma and muscle 3MH levels were well correlated, and an inverse curvilinear relationship between weight gain and plasma 3MH concentrations levels was observed. Plasma and muscle 3MH levels were significantly elevated in pair-fed birds, but 3MH levels in infected birds were increased by 30% over pair-fed birds. The results suggested that muscle breakdown, as assessed by plasma and muscle levels of 3MH, increased during the acute stage of Eimeria infection in turkey poults.     

Duration of immunity engendered by a single dose of a cold-adapted strain of Avian pneumovirus

The duration of immunity after a single dose of a cold-adapted strain of Avian pneumovirus (APV) was studied. Turkeys were vaccinated at 1 wk of age and challenged with virulent virus 3, 7, 10, and 14 wk later. Nonvaccinated groups were also challenged at the same times. No clinical signs were observed in the vaccinated birds after vaccination or after any challenge. No viral RNA was shed by the vaccinated birds after any challenge. The nonvaccinated birds shed viral RNA after all challenges. Avian pneumovirus-specific humoral antibodies were detected in the vaccinated birds until 14 wk after vaccination. The results of this preliminary study indicate that inoculation with a single dose of a cold-adapted strain of APV at 1 wk of age provides protection until 15 wk of age.     

Protection against neonatal Escherichia coli diarrhoea in pigs by vaccination of sows with a new vaccine that contains purified enterotoxic E. coli virulence factors F4ac, F4ab, F5 and F6 fimbrial antigens and heat-labile E. coli enterotoxin (LT) toxoid

The efficacy of a new vaccine against neonatal Escherichia coli diarrhoea in piglets containing purified F4ab, F4ac, F5 and F6 fimbriae and detoxified heat-labile toxin (LT) was tested in challenge experiments by the method described by the European Pharmacopoeia (3rd edn, EDQM, Council of Europe, Strasbourg, France). A group of 11 young sows from a herd without E. coli problems was vaccinated 6-8 and 2-4 weeks prior to expected farrowing and another group of nine young sows were non-vaccinated controls. Escherichia coli antibody titres were determined in serum samples taken from the sows before first vaccination and before farrowing and in colostrum samples. The newborn piglets were allowed to suckle colostrum from their mother immediately after birth. The piglets were marked with individually numbered ear tags. Approximately 12 h after birth, 118 piglets from vaccinated sows and 79 piglets from non-vaccinated control sows were challenged by oral instillation of 5 ml of a freshly prepared culture of one of the challenge strains [O8:K87:F4ab (LT+) or O149:K91:F4ac (LT+) or O9:K30:F5 or O9:K103:F6 respectively]. The challenge cultures contained as a mean 6.8x10(9) CFU/ml. After challenge the piglets were observed for 7 days and mortality and morbidity were recorded. Vaccinated sows developed significant levels of antibody titres in colostrum and serum. Control sows stayed at a low/seronegative level. The protective efficacy was excellent because 66.7-87.5% of the piglets from vaccinated sows remained without clinical signs after challenge. Only 0.0-28.0% of the piglets from non-vaccinated sows remained healthy and more than 47.1% of the piglets in this group died after challenge. It is concluded that the new vaccine is very effective in protection of piglets against neonatal E. coli diarrhoea.     

Immunogenicity of infectious bursal disease viruses in chickens.

Cross-protective properties of infectious bursal disease viruses (IBDVs) were studied. Viruses represented different subtypes of serotype 1, including recently isolated viruses (variants), and a serotype 2 virus. Chickens were vaccinated at 3 weeks of age with inactivated vaccines containing 10(5), 10(6), 10(7), or 10(8) mean tissue-culture infectious dose of a given virus and challenged 2 weeks later using either 10(2) or 10(3.5) mean embryo infectious dose (EID50) of either a standard virus or a variant serotype 1 virus. Protection was evaluated at 5 and 10 days post-challenge, based on gross and microscopic lesions, body weight, and bursa/body-weight ratios. The serotype 2 virus did not confer protection on birds challenged with the serotype 1 viruses. Vaccines made of variant viruses at the low doses protected chickens challenged with the high or low doses of either the standard or the variant viruses. Vaccines made of the standard or variant strains at low doses protected against high or low challenge doses of the standard strain. Vaccines made of the high dose of any of the standard strains protected chickens against the variant virus when the low challenge dose (10(2) EID50) was used, but not when the high challenge dose (10(3.5) EID50) was used. The lowest dose of the standard viruses vaccines required to confer protection against the variant virus varied depending on the strain. Results indicated that protection depended on the strain and dose of both the vaccine and challenge viruses and that the variant strains and standard strains share a common protective antigen(s).     

Evaluation of protection against Mycoplasma gallisepticum infection in chickens vaccinated with the F strain of M. gallisepticum

The effect of vaccination with the F strain of Mycoplasma gallisepticum (MG) on protection against challenge with a tylosin-resistant strain of MG was evaluated. White leghorn chickens vaccinated via eyedrop at 6 weeks of age were subsequently challenged with various dilutions of the tylosin-resistant MG strain, as were unvaccinated controls. Three days later, tracheal swabs were collected and cultured in medium with and without tylosin to distinguish between the vaccine and challenge strains. The mean infectious dose of the challenge strains was 3.8 log10 higher in the vaccinated group than in the controls, and the vaccinated group harbored fewer challenge organisms in the trachea. These findings suggest that the F strain of MG induces protection against infection with field strains of MG and that long-term vaccination with the F strain in multiple-age layer farms may result in replacement of field MG strains by the F strain.     

Safety of temperature sensitive mutant Mycoplasma gallisepticum vaccine

A temperature sensitive (ts) vaccine strain designated ts-11 was selected after exposure of a low passage culture of the immunogenic Australian field isolate (strain 80083) of Mycoplasma gallisepticum to 100 mg/ml of N-methyl-N-nitro-N-nitrosoguanidine. Viable counts (assayed as colour changing units (CCU)/25 microliters) of a thawed stock culture of ts-11 were typically log10 3 to log10 5 higher when incubated at 33 degrees C (the permissive temperature) than duplicate viable counts incubated at 39.5 degrees C (the restrictive temperature). Doses of approximately 2 x 10(7) CCU of ts-11 caused no gross lesions or loss of egg production when inoculated into the air sacs of susceptible chickens and no clinical or pathological signs of sinusitis when inoculated into the infraorbital sinuses of susceptible turkey poults, whereas the parent strain 80083 was demonstrably pathogenic. However, 1 of 10 poults inoculated intra-abdominally with approximately 2 x 10(7) CCU of ts-11 did show signs of mild airsacculitis. Eight-week-old pullets were vaccinated by eye drop with up to 1.4 x 10(7) CCU of ts-11 and simultaneously subjected to several stressful management practices, without apparent ill effects. Administration by coarse aerosol of 5 ml of ts-11 vaccine/25 day-old broilers, with or without 25 doses of infectious bronchitis virus vaccine caused no obvious signs of respiratory disease. The non virulent ts phenotype was maintained after 3 passages of strain ts-11 in chickens. Chickens vaccinated 3 weeks previously with ts-11 or with strain 80083 were placed in contact with susceptible chickens for a period of 2 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)     

鸡滑液囊支原体活疫苗(MS-H株)同时免疫对新支二联活疫苗免疫效力影响的调查

为了解新支二联活疫苗和鸡滑液囊支原体活疫苗之间有无相互影响,选用70只1日龄SPF鸡随机分为4组进行比对试验,其中:G1组20只,联合免疫新支二联苗活疫苗和鸡滑液囊支原体活疫苗(MS-H株);G2组20只,为新支二联活疫苗单独免疫组;G3组20只,为传染性支气管炎病毒(IBV)攻毒组;G4组10只,为空白对照组.免疫后...     

Salmonella enterica serovar typhimurium colonization of the crop in the domestic turkey: influence of probiotic and prebiotic treatment (Lactobacillus acidophilus and lactose)

Acute colonization of the crop of the domestic turkey by Salmonella enterica serovar typhimurium (ST) was examined. The influences of preharvest probiotic and prebiotic treatment with lactobaccilli and lactose on crop colonization with ST were also investigated. Prior to Salmonella challenge, poults received 2.5% lactose and Lactobacillus acidophilus (1.9 x 10(9) organisms/liter) in the only source of drinking water from 1 day old to termination. At 3-wk-old, turkey poults were challenged with ST (1.7 X 10(8) colony-forming units [CFU]/ml) before their natural nocturnal fast to determine the potential effects of supplementation on crop colonization when the crop was engorged and subsequently undergoing emptying. Crop ingesta and tissue were collected at time points 30 min and 4, 8, and 24 hr postchallenge and ST levels were determined. High levels of ST were detected in the crop. For instance, for the poults not receiving lactose or lactobacilli, 30 min after ST challenge, there were 4.4 x 10(7) CFU in the crop ingesta and 5.3 x 10(5) CFU in the crop wall. Ingesta ST levels dropped dramatically to 1.0 x 10(6) CFU after 4 hr as the crop emptied. Crop wall ST levels were steady during the nocturnal crop evacuation. Immunohistochemical staining demonstrated ST in close association with the crop epithelium. Treatment with lactose and L. acidophilus supplementation did not reduce ST colonization.     

Induction of humoral immune response and protective immunity in chickens against Salmonella enteritidis after a single dose of killed bacterium-loaded microspheres.

Formalin-inactivated Salmonella enteritidis phage type 4 strain 119/95 (SE) was encapsulated in biodegradable poly (DL-lactide co-glycolic acid) PLGA; (65:35) microspheres by a modified water-in-oil-in-water (w/o/w) double-emulsion solvent extraction/evaporation technique. These SE-loaded microspheres (SE-MS) were porous and spherical in shape with diameters of 0.4-10 microm and 20-80 microm in two preparations. SE-MS were subsequently used to vaccinate specific-pathogen-free chickens in a single dose in order to investigate the potency of a single-dose vaccination in inducing immune responses and protective immunity. In Experiment 1, 4-wk-old chickens that were vaccinated intramuscularly with 20-80-microm SE-MS generated long lasting (over 6 mo) and persistently high serum anti-SE immunoglobulin (Ig)G antibody response. In Experiment 2, 2-wk-old chickens were vaccinated orally with 0.4-10-microm or intramuscularly with 20-80-microm SE-MS and challenged with 10(9) colony-forming units of homologous SE strain at 6 wk postvaccination. When challenged intramuscularly, one each of the orally vaccinated (n = 10) and the intramuscularly vaccinated birds (n = 10) showed clinical signs and death, whereas all of the nonvaccinated control birds (n = 12) were sick and 11 of them were killed. When challenge was via oral route, 26.1% of cloacal swabs and 24.0% of organs (liver, spleen, and cecum) collected from orally vaccinated birds (n = 35) were positive for SE, comparable to 27.9% of feces and 18.7% of organs from the intramuscularly vaccinated birds (n = 35). These figures were significantly lower than those for nonvaccinated birds (n = 30) from which 59.3% of feces and 44.0% of organs tested SE positive (P < 0.05). The humoral immune response was also determined after vaccination with a single dose. The intramuscular vaccination elicited higher serum IgG response than oral administration, but the latter elicited a significant intestinal mucosal IgA antibody response. This is the first evidence that chickens vaccinated with killed SE-loaded PLGA microspheres, intramuscularly and orally in a single dose, developed systematic and local immune responses, thereby conferring protective immunity.     

An aerosolized Brucella spp. challenge model for laboratory animals

To characterize the optimal aerosol dosage of Brucella abortus strain 2308 (S2308) and B. melitensis (S16M) in a laboratory animal model of brucellosis, dosages of 10(3)-10(10) colony forming units (CFU) were nebulized to mice. Although tissue weights were minimally influenced, total CFU per tissues increased beginning at 10(6)-10(7) CFU dosages, with 10(9) CFU appearing to be an optimal dosage for S16M or S2308 aerosol delivery. At 12 weeks after vaccination with 10(7) CFU of B. abortus strain RB51 (SRB51) or saline (control), mice were challenged intraperitoneally (i.p.) (6.4 x 10(4) CFU) or via aerosol (1.76 x 10(9) CFU) with S2308. Mice vaccinated with SRB51 had reduced (P < 0.05) splenic, liver and lung colonization (total CFU and CFU/g) after i.p. challenge with S2308 as compared with control mice after i.p. S2308 challenge. Control and SRB51-vaccinated mice did not differ (P > 0.05) in splenic, liver or lung colonization after aerosol S2308 challenge. Failure to demonstrate vaccine protection was not because of a high aerosol challenge dosage as colonization of spleen and liver tissues was lower (P < 0.05) after aerosol challenge when compared with control mice after i.p. S2308 challenge.     

Protective immunity against Newcastle disease: the role of cell-mediated immunity

The role of cell-mediated immunity (CMI) in protection of birds from Newcastle disease was investigated by two different strategies in which only Newcastle disease virus (NDV)-specific CMI was conveyed without neutralizing antibodies. In the first strategy, selected 3-wk-old specific-pathogen-free (SPF) birds were vaccinated with either live NDV (LNDV), ultraviolet-inactivated NDV (UVNDV), sodium dodecyl sulfate-treated NDV (SDSNDV), or phosphate-buffered saline (PBS) (negative control) by the subcutaneous route. Birds were booster vaccinated 2 wk later and challenged with the velogenic Texas GB strain of NDV 1 wk after booster. All vaccinated birds had specific CMI responses to NDV as measured by a blastogenesis microassay. NDV neutralizing (VN) and hemagglutination inhibition (HI) antibody responses were detected in birds vaccinated with LNDV and UVNDV. However, birds vaccinated with SDSNDV developed antibodies that were detected by western blot analysis but not by the VN or HI test. Protection from challenge was observed only in those birds that had VN or HI antibody response. That is, birds with demonstrable CMI and VN or HI antibody response were protected, whereas birds with demonstrable CMI but no VN or HI antibody response were not protected. In the second strategy, birds from SPF embryos were treated in ovo with cyclophosphamide (CY) to deplete immune cells. The birds were monitored and, at 2 wk of age, were selected for the presence of T-cell activity and the absence of B-cell activity. Birds that had a significant T-cell response, but not a B-cell response, were vaccinated with either LNDV, UVNDV, or PBS at 3 wk of age along with the corresponding CY-untreated control birds. The birds were booster vaccinated at 5 wk of age and were challenged with Texas GB strain of NDV at 6 wk of age. All birds vaccinated with LNDV or UVNDV had a specific CMI response to NDV, VN or HI NDV antibodies were detected in all CY-nontreated vaccinated birds and some of the CY-treated vaccinated birds that were found to have regenerated their B-cell function at 1 wk postbooster. The challenge results clearly revealed that CY-treated birds that had NDV-specific CMI and VN or HI antibody responses to LNDV or UVNDV were protected, as were the CY-nontreated vaccinated birds. However, birds that had NDV-specific CMI response but did not have VN or HI antibodies were not protected from challenge. The results from both strategies indicate that specific CMI to NDV by itself is not protective against virulent NDV challenge. The presence of VN or HI antibodies is necessary in providing protection from Newcastle disease.