scholarly journals The immune response and protective efficacy of a potential DNA vaccine against virulent Pasteurella multocida

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Ahmed A. M. Yassein ◽  
Ayaat A. Teleb ◽  
Gamal M. Hassan ◽  
Zaki A. El Fiky
Keyword(s):  
Dna Vaccine ◽  
Pasteurella Multocida ◽  
Protective Efficacy ◽  
High Titer ◽  
Serum Antibody ◽  
Rflp Analysis ◽  
Elisa Test ◽  
Farm Animals ◽  
Serum Samples ◽  
Live Attenuated Vaccine

Abstract Background Pasteurella multocida is the main cause of several infections of farm animals, and the immunity gained from commercial vaccines is for the short term only and needs to be routinely administered, so work on new vaccines against virulent P. multocida is crucial. Results In this study, the OmpH gene was amplified from ten P. multocida strains, and the PCR products were sequenced and analyzed. The results of RFLP analysis of OmpH gene digested by MspI enzyme showed that all of ten strains examined possessed one restriction site and two fragments, 350 and 650 bp. The OmpH sequence of strain No. 10 was cloned into bacterial expression vector pUCP24, and the recombinant pUCP24-OmpH was expressed in E. coli DH5α. Serum samples obtained from the ELISA test from a group of vaccinated rats indicate that the antibodies were present at high titer in immunized rats and can be tested as a vaccine candidate with a challenge. Conclusions In rats infected with the DNA vaccine and inactivated vaccine, a significant increase in serum antibody levels was observed. In addition, the DNA vaccine provided the vaccinated rats with partial protection; however, the protective efficacy was greater than that offered by the live attenuated vaccine. This successful recombinant vaccine is immunogenic and may potentially be used as a vaccine in the future.

The Promotion of Peony Seed Proteolysis Product to the Immune Efficacy Induced by ptfA Gene DNA Vaccine of Avian Pasteurella multocida

2020 ◽  
Author(s):  
Qiang Gong ◽  
Zhenqi Du ◽  
Mingfu Niu ◽  
Cuili Qin
Keyword(s):  
Dna Vaccine ◽  
Pasteurella Multocida ◽  
Protective Efficacy ◽  
Stimulation Index ◽  
Serum Antibody ◽  
Peripheral Blood Lymphocytes ◽  
Vaccine Group ◽  
Live Attenuated Vaccine ◽  
Attenuated Vaccine ◽  
Immune Efficacy

Background: Avian Pasteurella multocida is one of the pathogens that affect the health of poultry. The protective efficacy of traditional attenuated vaccine is not ideal. In previous study, we prepared ptfA gene DNA vaccine of avian P. multocida. However, the protective effect of ptfA gene DNA vaccine was inferior to the attenuated vaccine. Therefore, it is necessary to improve the immune efficacy of avian P. multocida DNA vaccine, such as screening for novel adjuvant. Methods: In this study, the peony seed proteolysis product was gavaged to chickens before DNA vaccination or was added to the ptfA gene DNA vaccine as adjuvant. These vaccines were administered to chickens and the serum antibody, lymphocyte proliferation levels, IFN-g, IL-2, IL-4 and IL-6 concentrations secreted by peripheral blood lymphocytes were determined. After challenging with virulent avian P. multocida, survival time and protective efficacy was evaluated. Result: Following vaccination, no significant differences in antibody levels and concentrations of IL-4 among the DNA vaccine group, adjuvant-DNA vaccine group and gavaged group were observed. The stimulation index (SI) values, concentrations of IFN-g, IL-2 and IL-6 in adjuvant-DNA vaccine group and gavaged group were significantly higher than those in the DNA vaccine group. The protective efficacy of live attenuated vaccine group, DNA vaccine group, adjuvant-DNA vaccine group and gavaged group were 92%, 52%, 72% and 60%, respectively. This study has laid a foundation for the design and application of future DNA vaccines of P. multocida and DNA vaccine adjuvant.

scholarly journals Seroprevalence of Fasciola hepatica infection in cattle and sheep in the province of Kars, Turkey, as determined by ELISA

2014 ◽  
Vol 51 (2) ◽  
pp. 94-97 ◽  
Author(s):  
A. Akca ◽  
H. Gokce ◽  
N. Mor
Keyword(s):  
Host Species ◽  
Fasciola Hepatica ◽  
Elisa Test ◽  
Farm Animals ◽  
Serum Samples ◽  
Hepatica Infection ◽  
Significant Difference ◽  
Large Herd ◽  
Very High ◽  
Cattle And Sheep

Abstract This study aimed to determine the seroprevalence of Fasciola hepatica infection in cattle and sheep in the province of Kars, Turkey. Serum samples from 500 cattle and 540 sheep, collected from 15 randomly selected localities (villages) in the region, were tested for the presence of anti-F. hepatica antibodies using an in-house ELISA test with 98 % sensitivity and 96 % specificity. The seroprevalence of F. hepatica in the district was determined to be 66.6 % (333/500) in cattle and 93 % (502/540) in sheep. There was also a statistically significant difference in the rates of seropositivity between villages (each of which could be considered to be a large herd or flock), ranging from 0 % to 100 % in cattle and from 68 % to 100 % in sheep, P < 0.01). These findings show that F. hepatica infection is very common in the region; that, in contrast to results from abattoir which indicate a level of only 10 % prevalence, the exposure of farm animals to the infection in the region is in fact very high; and that the risk of acquiring the infection varies between the localities and the host species tested within the region.

scholarly journals Astragalus Polysaccharides Enhance the Immune Response to Newcastle Disease Virus Vaccination in Chickens

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Naji A .I . ◽  
◽  
Huda S.J ◽  
Keyword(s):  
Antibody Titer ◽  
Normal Saline ◽  
Sandwich Elisa ◽  
Serum Antibody ◽  
Elisa Test ◽  
Rt Pcr ◽  
High Concentration ◽  
Serum Samples ◽  
Astragalus Polysaccharide ◽  
Serum Antibody Titer

This study was conducted to evaluated the effects of Astragalus polysaccharide on immune responses of chickens immunized with NDV Lasota vaccine. One hundred chickens at one-day-old( Ross breed) were brought , on five day of age, the average maternal serum Abs was measured by hemaglutination inhibition (HI) test and the titer was 3.43 log, and then divided into five equal groups in each group twenty chicks and on day 10 of age the Astragalus polysaccharide high concentration (APSH) group were given 0.5 ml at 400mg /100ml , the Astragalus polysaccharide medium concentration (APSM )group were given 0.5 ml at 200mg /100ml, the Astragalus polysaccharide low concentration (APSL) group were given 0.5 ml at 100mg /100ml , Vaccine control (VC) group were given 0.5 ml normal saline only, Negative control (NC ) group were given normal saline only , all groups orally administrated for four days. At 14 day of age, all chickens with the exception of NC group , were vaccinated with ND Lasota by Intraocular and Intranasal methods. Four blood sample from all groups were aspirated from jugular vein at 0h (before vaccination),6h,12h and 24h(after vaccination ) for determine of Chicken IL-6(Interleukin 6) by RT-PCR. On day 20and 30 of age after vaccination , three chicken were sampled randomly from each group to detect specific serum Antibody titer of NDV by HI test. On days 1,7,14,21after vaccination, four serum Samples from the same of chicken to determines IgA level by sandwich ELISA test. The cellular and humeral response including the production of cytokine IL-6 ( by RT-PCR test) were measured on 0h(before vaccination),6h,12h and 24h( after vaccination )and IgA antibody ( by ELISA test ) were measured on 1d,7d,14d,and 21days after vaccination and serum antibody titer Specific to NDV( by HI test ) were measured at age 20, and 30 days after vaccination evaluated by series of experiments. Results revealed that all the polysaccharide groups were numerically increased in antibody levels, the expression of IL-6and IgA level ,but three parameter were significant (𝑃 < 0.05) in the APSH group compared to corresponding (VC) vaccinated group and( NC) non –vaccinated group. These results suggest that orally administered APS could significantly enhance the efficacy of NDV vaccination and has important implications for the further use of APS as a novel adjuvant.

scholarly journals Immune effects against influenza A virus and a novel DNA vaccine with co-expression of haemagglutinin- and neuraminidase-encoding genes

2009 ◽  
Vol 58 (7) ◽  
pp. 845-854 ◽  
Author(s):  
Weidong Zhang ◽  
Wanyi Li ◽  
Yan Li ◽  
Hong Li ◽  
Baoning Wang ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Dna Vaccine ◽  
Influenza A ◽  
Protective Efficacy ◽  
Survival Rates ◽  
Serum Antibody ◽  
Lethal Challenge ◽  
Vector Systems

The high variability of influenza virus causes difficulties in the control and prevention of influenza, thus seeking a promising approach for dealing with these problems is a hot topic. Haemagglutinin (HA) and neuraminidase (NA) are major surface antigens of the influenza virus, and provide effective protection against lethal challenges with this virus. We constructed a DNA vaccine (pHA-IRES2-NA) that co-expressed both HA and NA, and compared its protective efficacy and immunogenic ability with that of singly expressed HA or NA, or a mixture of the two singly expressed proteins. Our findings showed that both HA and NA proteins expressed by pHA-IRES2-NA could be detected in vivo and in vitro. The protection of DNA vaccines was evaluated by serum antibody titres, residual lung virus titres and survival rates of the mice. In the murine model, immunization of pHA-IRES2-NA generated significant anti-HA and anti-NA antibody, increased the percentage of CD8+ cells and gamma interferon-producing CD8+ cells and the ratio of Th1/Th2 (T helper) cells, which was comparable to the effects of immunization with HA or NA DNA alone or with a mixture of HA and NA DNA. All the mice inoculated by pHA-IRES2-NA resisted the lethal challenge by homologous influenza virus and survived with low lung virus titre. In addition, previous studies reported that co-expression allowed higher-frequency transduction compared to co-transduction of separated vector systems encoding different genes. The novel HA and NA co-expression DNA vaccine is a successful alternative to using a mixture of purified HA and NA proteins or HA and NA DNA.

scholarly journals Astragalus Polysaccharides Enhance the Immune Response to Newcastle Disease Virus Vaccination in Chickens

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Naji A.I ◽  
◽  
Huda S.J ◽  
Keyword(s):  
Antibody Titer ◽  
Normal Saline ◽  
Sandwich Elisa ◽  
Serum Antibody ◽  
Elisa Test ◽  
Rt Pcr ◽  
High Concentration ◽  
Serum Samples ◽  
Astragalus Polysaccharide ◽  
Serum Antibody Titer

This study was conducted to evaluated the effects of Astragalus polysaccharide on immune responses of chickens immunized with NDV Lasota vaccine. One hundred chickens at one-day-old( Ross breed) were brought , on five day of age, the average maternal serum Abs was measured by hemaglutination inhibition (HI) test and the titer was 3.43 log, and then divided into five equal groups in each group twenty chicks and on day 10 of age the Astragalus polysaccharide high concentration (APSH) group were given 0.5 ml at 400mg /100ml , the Astragalus polysaccharide medium concentration (APSM )group were given 0.5 ml at 200mg /100ml, the Astragalus polysaccharide low concentration (APSL) group were given 0.5 ml at 100mg /100ml , Vaccine control (VC) group were given 0.5 ml normal saline only, Negative control (NC ) group were given normal saline only , all groups orally administrated for four days. At 14 day of age, all chickens with the exception of NC group , were vaccinated with ND Lasota by Intraocular and Intranasal methods. Four blood sample from all groups were aspirated from jugular vein at 0h (before vaccination),6h,12h and 24h(after vaccination ) for determine of Chicken IL-6(Interleukin 6) by RT-PCR. On day 20and 30 of age after vaccination , three chicken were sampled randomly from each group to detect specific serum Antibody titer of NDV by HI test. On days 1,7,14,21after vaccination, four serum Samples from the same of chicken to determines IgA level by sandwich ELISA test. The cellular and humeral response including the production of cytokine IL-6 ( by RT-PCR test) were measured on 0h(before vaccination),6h,12h and 24h( after vaccination )and IgA antibody ( by ELISA test ) were measured on 1d,7d,14d,and 21days after vaccination and serum antibody titer Specific to NDV( by HI test ) were measured at age 20, and 30 days after vaccination evaluated by series of experiments. Results revealed that all the polysaccharide groups were numerically increased in antibody levels, the expression of IL-6and IgA level ,but three parameter were significant (𝑃 < 0.05) in the APSH group compared to corresponding (VC) vaccinated group and( NC) non –vaccinated group. These results suggest that orally administered APS could significantly enhance the efficacy of NDV vaccination and has important implications for the further use of APS as a novel adjuvant.

scholarly journals Enhancement of the Yersinia pestis EV NIIEG Vaccine Srain Immunogenic and Protective Activity under Cultivation with Azoxymer Bromide (Polyoxidonium)

2022 ◽  
Vol 20 (6) ◽  
pp. 12-19
Author(s):  
T. N. Shchukovskaya ◽  
A. Y. Goncharova ◽  
S. A. Bugorkova ◽  
O. M. Kudryavtseva ◽  
N. E. Shcherbakova ◽  
...  
Keyword(s):  
Yersinia Pestis ◽  
Vaccine Strain ◽  
Ribosomal Proteins ◽  
Protective Efficacy ◽  
Direct Action ◽  
Serum Antibody ◽  
Live Attenuated Vaccine ◽  
Maldi Tof ◽  
Morphological Studies ◽  
Characteristic Features

Background. The live-attenuated vaccine based on the Yersinia pestis strain EV line NIIEG is still used in Russia, providing protective efficacy against plague. Nevertheless, there is an urgent need for developing new ways to increase the immunogenicity of the Y. pestis EV NIIEG vaccine strain. In this study, the ability of direct action of immunoadjuvant azoximer bromide (polyoxidonium, PO) on the immunobiological properties of vaccine strain Y. pestis EV NIIEG during cultivation on a dense nutrient medium was evaluated. Materials & Methods. Y.pestis EV NIIEG, cultivated at 28 °С for 48 h on LB agar, Miller pH 7.2 ± 0.1 (Sigma-Aldrich, USA) with the addition of PO and without. MALDI-TOF mass-spectrometry was deployed for the obtainment of mass-spectra of ribosomal proteins from Y. pestis EV NIIEG cells on the MicroflexTM LT mass spectrometer (Bruker Daltonics, Germany). Protective efficacy was evaluated under subcutaneously challenge guinea pigs and mice BALB's with 400 LD50 doses of the Y. pestis 231, Y. pestis P-13268 Vietnam (MLD=5 CFU). Antibody titers to F1 in serum were determined using an ELISA. Results. The addition of the therapeutic concentration of PO in the cultivation medium induced a significant increase in the immunogenicity of Y. pestis EV NIIEG that resulted in enhancement of serum antibody levels against Y. pestis F1 antigen and several times the growth of protective efficacy in the bubonic plague model on two types of experimental animals. ImD50 of the vaccine strain Y. pestis EV NIIEG, cultivated with PO, was significantly (p < 0,05) lower in comparison to ImD50 for Y. pestis EV NIIEG in standard cultivation conditions. One year of storage at a temperature of 4 °С did not alter the protective properties of the vaccine strain Y. pestis EV NIIEG, cultivated with PO. Conclusions. Morphological studies confirmed the absence of influence PO introduction into the cultivation environment on the safety of the vaccine strain. MALDI-TOF MS profile of the Y. pestis EV NIIEG, cultivated with PO, had peaks characteristic features. The mass peak at m/z 3,061 was significantly down-regulated and new mass peaks at m/z 2,759, m/z 3,533 were determined. These changes are accompanied by the increase of Y. pestis EV NIIEG immunogenicity.

scholarly journals A network approach for provisional assay recognition of a Hendra virus antibody ELISA: test validation with low sample numbers from infected horses

2018 ◽  
Vol 30 (3) ◽  
pp. 362-369 ◽  
Author(s):  
Axel Colling ◽  
Ross Lunt ◽  
Jemma Bergfeld ◽  
Leanne McNabb ◽  
Kim Halpin ◽  
...  
Keyword(s):  
Test Performance ◽  
Serum Antibody ◽  
Relative Sensitivity ◽  
Elisa Test ◽  
Hendra Virus ◽  
Test Validation ◽  
Serum Samples ◽  
Antibody Elisa ◽  
Almost All ◽  
Positive Results

Obtaining statistically sound numbers of sera from Hendra virus (HeV)-infected horses is problematic because affected individuals usually die or are euthanized before developing a serum antibody response. As a consequence, test validation becomes a challenge. Our approach is an extension of OIE principles for provisional recognition and included 7 validation panels tested across multiple laboratories that provided estimates for test performance characteristics. At a 0.4 S/P cutoff, 16 of 19 sera from HeV-infected horses gave positive results in the HeV soluble G, indirect ELISA (HeVsG iELISA; DSe 84.2% [95% CI: 60.4–96.6%]); 463 of 477 non-infected horse sera tested negative (DSp 97.1% [95% CI: 95.1–98.4%]). The HeVsG iELISA eliminated almost all false-positive results from the previously used HeV iELISA, with marginally decreased relative sensitivity. Assay robustness was evaluated in inter-laboratory and proficiency testing panels. The HeVsG iELISA is considered to be fit for purpose for serosurveillance and international movement of horses when virus neutralization is used for follow-up testing of positive or inconclusive serum samples.

MONITORING STUDIES OF ARBOVIRUS INFECTIONS TRANSMITTED BY MOSQUITOES ON THE TERRITORY OF THE VOLGOGRAD REGION

2019 ◽  
pp. 60-66
Author(s):  
E.V. Molchanova ◽  
D.N. Luchinin ◽  
A.O. Negodenko ◽  
D.R. Prilepskaya ◽  
N.V. Boroday ◽  
...  
Keyword(s):  
Virus Antigen ◽  
Elisa Test ◽  
Serum Samples ◽  
Blood Sucking ◽  
Tick Borne Encephalitis ◽  
Volgograd Region ◽  
Two Samples ◽  
Probable Presence ◽  
California Serogroup ◽  
Tick Borne Encephalitis Virus

The paper presents data from the monitoring studies’ results of arbovirus infections transmitted by mosquitoes in the Volgograd region. West Nile virus antigen (WNV) in 9 samples, Tahyna virus in one sample, Batai virus in two samples were detected in the study of 110 samples of field material (blood-sucking mosquitoes) by ELISA test. Antibodies to WNV in 16.58 percent of the samples, to tick-borne encephalitis virus in 1.08 percent, to viruses of the California serogroup and Ukuniemi in 1.09 percent, to the virus Sindbis in 2.17 percent were detected as a result of the study of blood serum samples from donors in the Volgograd region. Thus, we obtained data on the probable presence of the Batai, Sindbis, Ukuniemi and Californian serogroup viruses along with the circulation of WNV on the territory of the Volgograd region.

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