Cross protection studies on Bordetella bronchiseptica in mice using an intracerebral challenge model.

Protective activities of heat-inactivated (60 degrees C for 30 min) merthiolate preserved Bordetella bronchiseptica and B. pertussis bacterins were compared in intraperitoneally immunized mice challenged intracerebrally (i.p./i.c.) or intraperitoneally (i.p./i.p.). In the i.p./i.c. assay (Kendrick test), a B. pertussis bacterin protected mice against challenge with B. pertussis 18-323, as well as against phase I cytotoxic and non-cytotoxic strains of B. bronchiseptica. A B. bronchiseptica bacterin, prepared from a phase I cytotoxic strain, gave protection against two phase I B. bronchiseptica strains, irrespective of their cytotoxin-production. A non-cytotoxic phase I strain of B. bronchiseptica elicited protection against the homologous strain only. Neither cytotoxic nor non-cytotoxic B. bronchiseptica strains protected mice challenged with B. pertussis 18-323. Vaccines prepared from phase III strains of B. bronchiseptica were not protective at all against any of the challenge strains. No such differences in the protective activities of the bacterins could be detected by the i.p./i.p. method. They seem to cross-protect equally well. The results indicate that the Kendrick test may be useful in testing potency of different B. bronchiseptica bacterins.     

A Salmonella abortusovis inactivated vaccine protects mice from abortion after challenge.

Two types of Salmonella abortusovis vaccines were prepared, one with aluminium hydroxide (vaccine A) and the other with water in oil (vaccine B) adjuvants. They were compared in a pregnant mouse model, aiming at protecting them from abortions after challenge with a virulent strain of S. abortusovis. The protection for vaccine A was from 74% to 77.6% and that for vaccine B from 71% to 79.6%. Abortions occurred 5-10 days post challenge and S. abortusovis was isolated from all aborted fetuses and from the liver and the spleen of their mothers at the end of the experiment (18 days post challenge). The presence of salmonella in the liver and the spleen of vaccinated non-pregnant but challenged mice was studied in a separate experiment. The bacterium was isolated from one out of 12 vaccinated mice 6 days post challenge as well as from the six controls.     

Effects of vaccine dose, virus challenge dose and interval from vaccination to challenge on protection of broiler chickens against Marek's disease virus challenge

OBJECTIVES: To examine the effects of varying the doses of turkey herpesvirus (HVT) vaccine and Marek's disease virus (MDV) challenge at two intervals after vaccination on the protection of chickens against challenge with MDV. DESIGN AND PROCEDURE: Experiment 1, a dose response study, consisted of 11 doses of HVT vaccine administered at hatch followed by challenge with 100 plaque forming units (pfu) of MDV 5 days post vaccination. Experiment 2, a 2 x 6 x 2 factorial design, included two HVT vaccine types, six different doses of HVT vaccine and 50 pfu and 200 pfu of MDV challenge 2 days post vaccination. All chickens were reared up to day 56 post challenge when all survivors were killed humanely. Dead and killed chickens were examined for gross MD tumours. RESULTS: Experiment 1 showed a significant positive linear relationship between dose of HVT vaccine and protective index in chickens challenged 5 days post vaccination. However the range of protective index observed was limited. In Experiment 2 neither HVT vaccine provided significant protection at any dose. There was no significant effect of vaccine type or MDV challenge dose on overall protection against challenge. Chickens challenged with 200 pfu of MDV had significantly higher mortality and MD incidence than those with 50 pfu. CONCLUSIONS: HVT vaccine dose had a significant impact on protective index, but vaccination to challenge interval appeared to have greater impact on the protective efficacy of vaccination. A fourfold increase in challenge dose increased mortality rate and incidence of MD.     

Immunogenicity of replication-deficient vesicular stomatitis virus based rabies vaccine in mice

BackgroundRabies is a viral disease that causes severe neurological manifestations both in humans and various mammals. Although inactivated and/or attenuated vaccines have been developed and widely used around the world, there are still concerns with regard to their safety, efficacy, and costs.ObjectiveAs demand has grown for a new rabies vaccine, we have developed a new vesicular stomatitis viruses (VSVs) based rabies vaccine that replaces glycoproteins with rabies virus (RABV) glycoprotein (GP), or so-called VSV/RABV-GP.MethodsVSV/RABV-GP production was measured by sandwich ELISA. The generation of VSV/RABV-GP was evaluated with GP-specific antibodies and reduced transduction with GP-specific neutralizing antibodies. Virus entry was quantified by measuring the luciferase levels at 18-h post-transduction. BALB/c mice (three groups of six mice each) were intraperitoneally immunized with PBS, RABA, or VSV/RABV-GP at 0 and 14 days. At 28 days post-immunization serology was performed. Statistical significance was calculated using the Holm–Sidak multiple Student’s t test.ResultsMice immunized with VSV/RABV-GP produced IgM and IgG antibodies, whereas IgM titers were significantly higher in mice immunized with VSV/RABV-GP compared to inactivated RABV. The secretion profiles of IgG1 and IgG2a production suggested that VSV/RAVB-GP induces the T helper cell type-2 immune bias. In addition, the average (±SD; n = 3) serum neutralization titers of the inactivated RABV and VSV/RABV-GP groups were 241 ± 40 and 103 ± 54 IU/mL, respectivelyConclusionOur results confirm that VSV/RABV-GP could be a new potential vaccination platform for RABV.     

Immunization of foxes by the intestinal route using an inactivated rabies vaccine.

Approximately 30% of foxes given two doses of an inactivated rabies antigen delivered directly into the intestinal tract developed an immune response as measured by rabies serum neutralizing antibodies. Seven of ten previously immunized foxes showed an anamnestic response following a booster dose of inactivated rabies antigen delivered to the intestinal lumen. Stomach and particularly intestinal contents were destructive to rabies antigen and virus. This effect could be partially neutralized in vitro by the addition of Questran and soybean trypsin inhibitor. Small enteric coated tablets fed to foxes in a hamburger bolus remained in the stomach for up to 13 hours and therefore would provide a poor vehicle for the delivery of antigen to the intestinal tract.     

Studies of ERA/BHK-21 rabies vaccine in skunks and mice.

ERA rabies vaccine virus grown in BHK-21 13S cells (ERA/BHK-21) and street rabies virus were titrated in mice by intracerebral, intranasal and intramuscular inoculation. Mice were also given undiluted ERA/BHK-21 in baits. Skunks were given undiluted ERA/BHK-21 in baits and by intramuscular, intranasal and intestinal inoculation. Virus neutralizing antibody titers against rabies virus were measured over a three month observation period. The surviving skunks were challenged by intramuscular inoculation with rabies street virus from a skunk salivary gland suspension. When titrated in mice, ERA/BHK-21 had titers of 10(7.0), 10(5.2) and 10(3.9) median lethal doses per mL by the intracerebral, intranasal and intramuscular routes, respectively. All skunks (8/8) inoculated intranasally developed paralytic rabies by 12 days after exposure to ERA/BHK-21 virus. None of the skunks that developed vaccine-induced rabies had infectious virus in the submandibular salivary glands. Vaccine-induced rabies also occurred in 1/8 skunks in the intramuscularly inoculated group and in 1/8 in the intestinally inoculated group. The survival rates of challenged skunks in the various groups were as follows: intramuscular, 7/7; intestinal, 2/7; bait, 0/8; and control, 0/8. These results indicate that ERA/BHK-21 virus has a significant residual pathogenicity in mice and in skunks by some routes of inoculation. Skunks given vaccine intramuscularly were protected against challenge, while those skunks given the vaccine in baits were not.     

The effect of route of inoculation and challenge dosage on Riemerella anatipestifer infection in Pekin ducks (Anas platyrhynchos)

Riemerella anatipestifer is a gram-negative bacteria that can cause disease in a wide variety of wild and domesticated birds, especially waterfowl. The infection can be peracute, acute, or chronic. Although various routes of transmission have been proposed, to date, there is little information on the effects of route of transmission and challenge dosage on R. anatipestifer infection. Hence, the objective of this study was to determine the effect of route of inoculation and challenge dosage on R. anatipestifer infection and pathology. To achieve this objective, one hundred forty-seven 14-day-old white Pekin ducks (Anas platyrhynchos) were equally divided into 13 experimental groups (12 challenge and 1 control group). Each challenge group had 11 ducks. The control group had 15 ducks. Four routes of inoculation were evaluated (intranasal, oral, subcutaneous, and intravenous). Three dosage levels were evaluated for each inoculation route (10(2), 10(4), and 106 colony forming units [CFU]/ml). At the 106 CFU/ml dosage level, mortality was most associated with the subcutaneous (91%) and intravenous (82%) routes, followed by the nasal (18%) and oral (9%) routes. A unique pathologic lesion was found in the bursa of Fabricius and spleen of affected birds. Within the spleen and bursa of Fabricius, there were varying degrees of lymphoid depletion and necrosis within the cortical and medullary regions. These pathologic lesions have not been previously reported in ducks with R. anatipestifer infection.     

Cross protection of mice and swine given live-organism vaccine against challenge exposure with strains of Erysipelothrix rhusiopathiae representing ten serovars

Mice and swine vaccinated (subcutaneous inoculation) with live acriflavine-fast attenuated Erysipelothrix rhusiopathiae, strain Koganei 65-0.15 (serovar 2), were challenge exposed with 10 strains of E rhusiopathiae pathogenic for swine; the latter strains comprised serovars 9 and 10 and other previously undetermined. Vaccinated mice did not die after they were challenge exposed (subcutaneous inoculation) with serovars 4, 6, 7, 8, 9, 10, 15, 16, or N, but vaccinated mice challenge exposed with strain 2553 (serovar 20) had 30% mortality. Nonvaccinated control mice died after they were challenge exposed with all serovars tested. One of 2 vaccinated swine challenge exposed (intradermal inoculation) with each of strains 911 (serovar 8), 2179 (serovar 10), or 2553 developed localized urticarial lesion at the site of intradermal inoculation. Vaccinated swine challenge exposed with serovars 4, 6, 7, 9, 15, 16, or N did not have clinical signs of acute swine erysipelas. Nonvaccinated control swine developed localized lesions at the site of intradermal challenge inoculation.     

猪口蹄疫O型灭活疫苗PD50效检攻毒剂量试验

采用PD50效检方法,对0304041-2批猪口蹄疫O型灭活疫苗(I型苗)进行不同攻毒剂量的PD50 检测.结果表明,分别用500 ID50 、1 000 ID50 、2 000 ID50 和4 000 ID50 的病毒进行攻毒,疫苗每头份分别含5.2、3.6、2.1和1.0个PD50 .该项试验的攻毒毒株ORMF8的ID50为10-7.0/2 mL,MID为10 -6.0 /2 mL.     

Cross protection in mice and swine immunized with live erysipelas vaccine to challenge exposure with strains of Erysipelothrix rhusiopathiae of various serotypes

Mice and swine immunized subcutaneously with live vaccine prepared from acriflavine-fast attenuated Erysipelothrix rhusiopathiae, strain Koganei (serotype 2), were challenge exposed to virulent strains of E rhusiopathiae of various serotypes. Vaccinated mice did not die after challenge exposure to serotypes 1a, 1b, 2, 3, 5, 6, 7, 8, 9, 11, 12, 15, 16, 18, 19, 21, or N, but 20% to 30% mortality occurred in vaccinated mice challenge exposed to serotypes 10, 14, 20, or 22. Nonvaccinated control mice died after challenge exposure to all serotypes tested. Vaccinated swine challenge exposed to strain 14B (serotype 9) or strain 2179 (serotype 10) developed localized urticarial lesions at the site of intradermal exposure. Vaccinated swine challenge exposed to serotypes 1a, 1b, 2, 5, 8, 11, 12, 18, 19, or 21 did not have clinical signs of acute erysipelas. Nonvaccinated control swine developed acute generalized erysipelas or localized lesions at the site of intradermal exposure.     

猪圆环病毒2型核酸疫苗的小鼠免疫保护试验

为了研究猪圆环病毒2型(PCV2)核酸疫苗在小鼠攻毒试验中的免疫保护效果,以PCV-2 GXWZ-1株为模板,扩增出ORF2基因及其截短基因8个片段(A(ORF2)、B(51-100aa)、C(101-150aa)、D(181-235aa)、E(151-200aa)、F(51-150aa)、G(101-235aa)、H(51-235aa)),将其插入到pcDNA3.0载体中,构建出真核表达质粒,并将其转染至PK-15细胞,用间接免疫荧光试验检测其瞬时表达情况。将试验小鼠随机分成9组,其中免疫组7组,阴阳性对照各1组,将纯化的真核表达质粒对小鼠进行组合免疫;二免后,用经处理过的PCV-2 GXWZ-2株阳性病料悬液腹腔注射免疫组和非免疫对照组小鼠,阴性对照组用生理盐水腹腔注射;其后进行体重记录、病理切片制作及PCR检测。结果表明:共有6个真核表达质粒成功在PK-15细胞中表达。在攻毒后的3周内,阳性对照组小鼠PCR诊断均为阳性;免疫组中,部分组小鼠在攻毒后第1周或在第2周为阳性,到第3周时各免疫组小鼠全部为阴性;阴性对照组始终为阴性。免疫组在病理保护学方面明显优于非免疫对照组,非免疫对照组的体重增长速率略低于免疫组和阴性对照组。由此可见猪圆环病毒2型核酸疫苗在小鼠攻毒试验中有明显的保护作用。     

Cross-clade protection against HPAI H5N1 influenza virus challenge in BALB/c mice intranasally administered adjuvant-combined influenza vaccine

The avian H5N1 influenza virus has the potential to cause a new pandemic. The increasing number of recent outbreaks of highly pathogenic avian influenza H5N1 in birds and humans emphasizes the urgent need to develop a potent H5N1 vaccine. Here, we studied the immunogenicity and protective effect of a vaccine prepared from H5N1 inactivated whole virus. This vaccine was intranasally co-administered in mice with phosphate buffered saline, recombinant cholera toxin B subunit (rCTB), cholera toxin (CT), rCTB containing a trace amount of holotoxin (rCTB/CT), polyinosinic:polycytidylic acid double-stranded RNA (polyI:C), or MF59 as an adjuvant. Intranasal administration of H5N1 inactivated whole virus vaccine with rCTB, CT, rCTB/CT, polyI:C, and MF59 elicited an immunological response with both secretory IgA (sIgA) in nasal, lung, and vaginal lavage, and IgG antibody in serum, showing protective immunity against lethal H5N1 infection. Cross-clade protection was also observed in animals immunized with a vaccine derived from Anhui/01/2005(H5N1) with rCTB, CT, rCTB/CT, polyI:C, or MF59 as adjuvants that were subsequently challenged with the A/OT/SZ/097/03 influenza strain.     

Effect of FMD vaccine antigen payload on protection, sub-clinical infection and persistence following needle challenge in sheep

The relationship of Foot-and-Mouth Disease (FMD) virus antigen payload and single and double vaccinations in conferring protection against virus challenge in sheep was studied. Sheep vaccinated with half the cattle dose (1 ml) containing 15 and 3.75 μg of FMDV antigen with or without booster resisted virulent challenge on 21 days post vaccination or 7 days post booster. FMDV RNA could be detected in nasal secretions in 26% of vaccinated sheep (10^3.12 to 10^3.82 viral RNA copies) on day 35 post challenge. No live virus could be isolated after 5 days post challenge indicating that the risk of transmission of disease was probably very low. The finding showed that vaccines containing antigen payload of 1.88 μg may prevent or reduce the local virus replication at the oropharynx and shedding of virus from nasal secretions and thereby reduce the amount of virus released into the environment subsequent to exposure to live virus. Sheep with no vaccination or with poor sero conversion to vaccination can be infected without overt clinical signs and became carriers.     

The effect of route and dosage of immunization on the serological response to a Pasteurella haemolytica and Haemophilus somnus vaccine in feedlot calves

The effect of route and dosage of administration on the serological response to a vaccine containing genetically attenuated leukotoxin of Pasteurella haemolytica combined with bacterial extracts of P. haemolytica and Haemophilus somnus (Somnu-Star Ph, Biostar Inc., Saskatoon, Saskatchewan) was evaluated in a controlled field trial in 301 feedlot calves. Vaccination of calves on arrival at the feedlot with Somnu-Star Ph significantly (p < 0.05) increased P. haemolytica and H. somnus serum antibody titers and reduced bovine respiratory disease (BRD) morbidity. A single subcutaneous vaccination with Somnu-Star Ph was as effective in stimulating a humoral antibody response and in reducing BRD morbidity as double vaccination by the intramuscular or the subcutaneous route. Furthermore, there were no swellings or adverse reactions observed with either subcutaneous or intramuscular administration of Somnu-Star Ph.

These results suggest that feedlot calves can be immunized subcutaneously once on arrival with Somnu-Star Ph. Double vaccination was of no added value in this trial, because the majority of BRD morbidity occurred prior to revaccination fourteen days postarrival. Additional larger-sized field trials are needed to monitor the duration of immunity following vaccination and to test the effect of route and dosage of vaccination on mortality.