Protective immune response of oral rabies vaccine in stray dogs, corsacs and steppe wolves after a single immunization.

In this study the safety and protective immunity of an oral rabies vaccine, based on the live, modified rabies virus strain VRC-RZ2, was examined in stray dogs (Canis Sp.), corsacs (Vulpes corsac) and steppe wolves (Canis lupus campestris). In the safety group (dogs, n=6; corsacs, n=3; wolves, n=3) which was vaccinated with a 10-times field dose/animal, no animals showed any signs of disease or changes in behavior or appetite during the period of clinical observation, similar to the animals in the negative control group. Saliva samples taken from animals prior and post (5th and 10th days) vaccination failed to demonstrate rabies virus antigen. Observations of immunogenicity in vaccinated carnivores (dogs, corsacs and wolves) during a 180 day period showed the titers of virus neutralizing antibodies (VNA) in the blood sera of vaccinated dogs to be within 0.59-1.37 IU/mL. On 14 days post vaccination (dpv), all the wild carnivores had detectable levels of neutralizing antibodies, with mean titers ranging from 0.50 ± 0.07 IU/mL (for wolves) to 0.59 ± 0.10 IU/mL (for corsacs). Weeks after vaccination, all the vaccinated wolves and corsacs had higher levels of neutralizing antibodies: 0.70 ± 0.10 - 0.71 ± 0.08 IU/mL at 30 dpv, 1.06 ± 0.08 - 1.28 ± 0.21 IU/mL at 60 dpv and 0.41 ± 0.09 - 047 ± 0.06 at 180 dpv. The highest level of VNA (˃1.0 IU/ml) was detected at 60 dpv, in all vaccinated animals. After challenge all vaccinated dogs remained healthy for 180 days. Control animals (unvaccinated dogs) developed symptoms of rabies on day 6 post administration of a virulent virus and died of rabies on days 11-13. Of note, the VNA titers in all the wild carnivores (corsacs and wolves) immunized with VRC-RZ2 were higher than 0.5 IU/ml (0.59 ± 0.11 IU/ml), even as early as 14 days post vaccination. These, presumably protective, titers of antibodies to rabies virus were present in the dogs and wild carnivores examined in this study for at least 180 days.

[Genetic stability of the HA, NA, and NS genes of the recombinant vector virus Flu-NS1-124-Omp16 (H5N1) expressing the brucellar gene].

The recombinant strain Flu-NS1-124-Omp16 (H5N1) of the influenza virus expressing the brucellar Omp16 gene was constructed on the basis of the technology of reverse genetics for the purpose of developing vector anti-brucellosis vaccine. The obtained recombinant strain is a genetically stable construction. This stability is confirmed by the comparative analysis of the nucleotide sequences of the HA, NA, and NS genes of the recombinant vector virus Flu-NS1-124-Omp16 (H5N1) expressing the Omp16 gene of the Brucella abortus (GenBank: AAA59360.1). The comparative analysis showed that the nucleotide sequence of the NS gene of the first and the fifth passage level of the Flu-NS1-124-Omp16 (H5N1) virus corresponded for 100% to the initial part of 12AAS2TC_124 Omp16g containing the chimera NS1-124-Omp16 in the composition of DNA (deoxyribonucleic acid) plasmids pHW2000. Total identity with HA and NA genes of the strain A/AstanaRG/6:2/2009 (H5N1) was shown by the comparative analysis of the nucleotide sequences of HA and NA genes of the first and the fifth passage level of the recombinant strain Flu-NS1-124-Omp16 (H5N1). The recombinant vector virus Flu-NS1-124-Omp16 (H5N1) expressing the brucella Omp16 gene maintains the genetic stability during 5 passages in 10-day developing chicken embryos.

Comparative evaluation of effectiveness of IAVchip DNA microarray in influenza A diagnosis.

The paper describes comparative evaluation of IAVchip DNA microarray, reverse transcription PCR (RT-PCR), and real-time RT-PCR versus virus isolation in chicken embryos and shows their diagnostic effectiveness in detection and subtyping of influenza A virus. The tests were evaluated with use of 185 specimens from humans, animals, and birds. IAVchip DNA microarray demonstrates higher diagnostic effectiveness (99.45%) in early influenza A diagnosis as compared to the real-time PCR (98.38%) and RT-PCR (96.22%), thus showing its clear superiority. Diagnostic sensitivity of IAVchip DNA microarray (100%) exceeds the same of RT-PCR (95.95%) and real-time RT-PCR (97.96%) in the range of estimated confidence intervals. IAVchip DNA microarray and real-time RT-PCR displayed equal diagnostic specificity (98.85%), while diagnostic specificity of RT-PCR was 96.40%. IAVchip DNA microarray has an advantage over the other tests for influenza A diagnosis and virus identification as a more rapid method that allows performing simultaneous detection and subtyping of about tens of specimens within one experiment during 8-10 hours. The developed IAVchip DNA microarray is a general test tool that enables identifying simultaneously 16 hemagglutinin (HA) and 9 neuraminidase (NA) subtypes of influenza A virus and also to screen the influenza A viruses from humans, animals, and birds by M and NP genes.

A New Intranasal Influenza Vector-Based Vaccine TB/FLU-04L Against Tuberculosis: Preclinical Safety Studies.

A recombinant vector vaccine TB/FLU-04L for the prevention of tuberculosis was developed in RIBSP CS MES RK and SRII. The vaccine is based on the attenuated influenza strain Flu NS106/ESAT-6_Ag85A expressing mycobacterial antigens Esat-6 and Ag85A. This research aimed to conduct pre-clinical safety studies of the vaccine as one of the basic and mandatory stages in the development and introduction of immunobiological preparations. The studies were performed at the research centers of the Republic of Kazakhstan and the Russian Federation.The experiment was conducted on ferrets, monkeys, and rabbits. The TB/FLU-04L vaccine was administered intranasally (7.5 lg TCID50/animal). The clinical signs, body weight, temperature, hematological parameters, and local irritant effects were monitored throughout the study. The results of the study demonstrated the safety of the TB/FLU-04L intranasal vector vaccine against tuberculosis since its administration in laboratory animals led to no adverse effects in any of the monitored parameters. No influenza A virus particles were isolated from samples of nasal washes.

Detection and Genetic Characterization of Lineage IV Peste Des Petits Ruminant Virus in Kazakhstan.

Peste des petits ruminant (PPR) is endemic in many Asian countries with expansion of the range in recent years including across China during 2013-2014 (OIE, 2014). Till the end of 2014, no cases of PPR virus (PPRV) were officially reported to the Office Internationale des Epizooties (OIE) from Kazakhstan. This study describes for the first time clinicopathological, epidemiological and genetic characterization of PPRV in 3 farm level outbreaks reported for the first time in Zhambyl region (oblast), southern Kazakhstan. Phylogenetic analysis based on partial N gene sequence data confirms the lineage IV PPRV circulation, similar to the virus that recently circulated in China. The isolated viruses are 99.5-99.7% identical to the PPRV isolated in 2014 from Heilongjiang Province in China and therefore providing evidence of transboundary spread of PPRV. There is a risk of further maintenance of virus in young stock despite vaccination of adult sheep and goats, along livestock trade and pastoral routes, threatening both small livestock and endangered susceptible wildlife populations throughout Kazakhstan.

Duration of the protective immune response after prime and booster vaccination of yearlings with a live modified cold-adapted viral vaccine against equine influenza.

We previously created a live vaccine against equine influenza based the new reassortant cold-adapted (Ca) strain A/HK/Otar/6:2/2010. The live vaccine contains surface proteins (HA, NA) from the wild-type virus A/equine/Otar/764/2007 (Н3N8; American Lineage Florida Clade 2), and internal proteins (PB2, PB1, PA, NP, M, NS) from the attenuated Ca donor virus A/Hong Kong/1/68/162/35CA (H3N2). To determine the safety and duration of the protective immune responses, 90 yearlings were intranasally vaccinated in single mode, double mode at an interval of 42 days (10(7.0) EID50/animal for both vaccinations), or with PBS (control group). Ten animals from each group were challenged with the homologous wild-type virus A/equine/Otar/764/07 (Н3N8) at 1, 2, 3, 4, 5, 6, 9 and 12 months after vaccination. Similarly, 10 animals from each group were challenged with the heterologous wild-type virus A/equine/Sydney/2888-8/07 (Н3N8; American Lineage Florida Clade 1) 12 months after vaccination. The vaccine was completely safe, and single intranasal vaccination of yearlings was capable of inducing statistically significant (from P=0.03 to P<0.0001) clinical and virological protection against the homologous virus; however, only double mode vaccination generated significant (from P=0.02 to P<0.0001) protection against the heterologous virus at 12 months (observation period). Interestingly, this vaccine enables the differentiation of infected and vaccinated animals. On this basis of this study, we recommend double intranasal administration of this vaccine at an interval of 42 days in veterinary practice.

Safety and immunogenicity of a novel cold-adapted modified-live equine influenza virus vaccine.

OBJECTIVE: To design and evaluate the safety and immunogenicity of a modified-live vaccine to prevent equine influenza virus (EIV) infection based on the novel reassortant cold-adapted strain A/HK/Otar/6:2/2010. METHODS: Surface proteins (HA, NA) from the wild-type strain A/equine/Otar/764/2007 (H3N8) and internal proteins (PB2, PB1, PA, NP, M, NS) from the attenuated cold-adapted donor strain A/Hong Kong/1/68/162/35CA (H3N2) were included in the vaccine. Horses were administered 10(9.2) EID50 /mL of the modified-live vaccine or saline solution using a nasal spray. The clinical condition of the animals was assessed throughout the study and nasopharyngeal swabs were collected for virus titration. Two yearlings in each group were euthanased on day 5 post vaccination (PV) for histological examination and measurement of viral titres in the organs. Serum samples and nasal secretions were collected to evaluate serological response. Lymphoproliferation after restimulation in vitro was determined to evaluate cell-mediated immunity. To evaluate the protective capacity of the vaccine, the yearlings in both groups were challenged with the wild-type virus at 28 days PV and their clinical condition and serological response was evaluated. Nasal swabs were collected to assess viral shedding from the upper respiratory tract. RESULTS: Single intranasal administration of a modified-live EIV vaccine caused no adverse effects and vaccinated yearlings and pregnant mares did not form detectable levels of antibodies by days 7, 14 and 28 PV, as indicated by the HI reaction and ELISA. Secretory antibodies could be detected on day 7 and reached maximal levels on day 14 PV. In vitro studies showed that the yearlings and pregnant mares both formed a cell-mediated immune response by day 14 PV. The vaccine protected yearlings against challenge with wild-type virus. We conclude that single intranasal administration of the modified-live EIV vaccine was safe in the yearlings and pregnant mares that we treated, and was immunogenic and protective in the yearlings.