Non-vaccinated Animals Research Articles

The effect of vaccination on the live weight of chickens and the absolute weight of the spleen and gland

Choosing optimal chicken immunization programs is an urgent problem in poultry farming. Studying the morphology of the organs of the chickens' immune system allows you to objectively assess the effectiveness of prevention methods. The article provides data on the effect of multiple vaccinations on the live weight of chickens and the absolute weight of immune defense organs: the spleen and gland of chickens aged 1, 15, 25, 50, 75, 100, and 120 days. One of the leading indicators of the growth of young birds is the dynamics of live weight under the influence of external factors. Identical technological conditions at the beginning of the experiment and the selection of livestock according to the principle of analogs in groups made it possible to assess the effect of vaccination on the chickens' bodies based on live weight indicators. According to our research results, it was established that in chickens of 100- and 120-day-old age, with multiple vaccinations, there is a probable increase in live weight indicators compared to the control group of animals. Our research showed that by 25 days in chickens (vaccinated six times), the hands of the absolute mass of the spleen practically did not change. At the same time, in 25-, 100-, and 120-day-old chickens, there is an increase in the absolute mass of the spleen and the gland at 15- and 50-day-old compared to the indicators of non-vaccinated animals. This indicates the implementation of immune reactions in the studied organs of immune protection due to increased migration of T- and B-lymphocytes to them. In addition, if age periods are compared, a decrease in the absolute mass of the Garder gland was found in 75-day-old chickens compared to 50-day-old chickens, which indicated the involutional processes of this gland. The maximum parameters of the gland's absolute mass were found at 50 days in chickens of both the vaccinated and non-vaccinated groups. Thus, in vaccinated chickens, this indicator of the gland was 0.095 g. Organometric changes in the spleen and gland of vaccinated chickens can be one of the criteria for diagnosing poultry immunodeficiencies.

Novel Proteoliposome-Based Vaccine against E. coli: A Potential New Tool for the Control of Bovine Mastitis.

Simple SummaryMastitis is a highly prevalent disease in dairy cattle, affecting animal welfare and generating economic losses for the dairy industry. Control measures for coliform mastitis are limited, due to the constant exposure of the teat to bacteria and the emergence of antimicrobial-resistant bacteria, making vaccination an important strategy for control of mastitis. However, currently available vaccines show limited efficacy, which could be attributed to inactivation processes that alter the antigenic preservation of the vaccines. The aim of this study was to assess the efficacy of a novel vaccine against mastitis using proteoliposomes obtained from E. coli in a murine model of coliform mastitis. We demonstrated that the proteoliposome vaccine was safe, immunogenic and effective against an experimental model of E. coli mastitis, decreasing bacterial count and tissue damage. This proteoliposome vaccine is a potential new tool for prevention of mastitis.Escherichia coli is an important causative agent of clinical mastitis in cattle. Current available vaccines have shown limited protection. We evaluated the efficacy of a novel vaccine based on bacterial proteoliposomes derived from an E. coli field strain. Female BALB/c mice were immunized subcutaneously with two doses of the vaccine, 3 weeks apart. Between days 5 and 8 after the first inoculation, the females were mated. At 5–8 days postpartum, the mice were intramammary challenged with the same E. coli strain. Two days after bacterial infection, mice were euthanized, and the mammary glands were examined and removed to evaluate the efficacy and safety of the vaccine as well as the immune response generated by the new formulation. The vaccinated mice showed mild clinical symptoms and a lower mammary bacterial load as compared to non-vaccinated animals. The vaccination induced an increase in levels of IgG, IgG1 and IgG2a against E. coli in blood and mammary glands that showed less inflammatory infiltration and tissue damage, as compared to the control group. In summary, the vaccine based on bacterial proteoliposomes is safe, immunogenic, and effective against E. coli, constituting a new potential tool for mastitis control.

Evolution of Swine Influenza Virus H3N2 in Vaccinated and Nonvaccinated Pigs after Previous Natural H1N1 Infection.

Swine influenza viruses (SIV) produce a highly contagious and worldwide distributed disease that can cause important economic losses to the pig industry. Currently, this virus is endemic in farms and, although used limitedly, trivalent vaccine application is the most extended strategy to control SIV. The presence of pre-existing immunity against SIV may modulate the evolutionary dynamic of this virus. To better understand these dynamics, the viral variants generated in vaccinated and nonvaccinated H3N2 challenged pigs after recovery from a natural A(H1N1) pdm09 infection were determined and analyzed. In total, seventeen whole SIV genomes were determined, 6 from vaccinated, and 10 from nonvaccinated animals and their inoculum, by NGS. Herein, 214 de novo substitutions were found along all SIV segments, 44 of them being nonsynonymous ones with an allele frequency greater than 5%. Nonsynonymous substitutions were not found in NP; meanwhile, many of these were allocated in PB2, PB1, and NS1 proteins. Regarding HA and NA proteins, higher nucleotide diversity, proportionally more nonsynonymous substitutions with an allele frequency greater than 5%, and different domain allocations of mutants, were observed in vaccinated animals, indicating different evolutionary dynamics. This study highlights the rapid adaptability of SIV in different environments.

Experimental reproduction of histomonosis caused by Histomonas meleagridis genotype 2 in turkeys can be prevented by oral vaccination of day-old birds with a monoxenic genotype 1 vaccine candidate

Histomonosis (syn. blackhead disease) is caused by the protozoan parasite Histomonas meleagridis and can result in high mortality in turkey flocks, a situation driven by the limitation of prophylactic and therapeutic interventions. Multi-locus sequence typing confirmed the existence of two genotypes, with the vast majority of reported histomonosis outbreaks being caused by genotype 1 in contrast to only a few detections of genotype 2.For the first time, genotype 2 of H. meleagridis was successfully isolated from an outbreak of histomonosis in a flock of 5-week-old turkeys and a clonal culture was established. Using this culture, an experimental infection was performed in naïve turkeys. The animal trial reflected the observations from the field outbreak and coincided with a previously reported case of histomonosis caused by genotype 2, albeit no mortality was observed in the infected birds whereas 17.1% mortality was noticed in the field outbreak from appearance of disease until slaughter. Post mortem investigations demonstrated that lesions were restricted to the caeca in the field outbreak and the experimental trial.In parallel with the experimental reproduction of pathological changes, an oral vaccination of day-old turkeys with a monoxenic genotype 1 vaccine was carried out to determine efficacy against a genotype 2 challenge. Successful vaccine uptake was characterized by the presence of the vaccine in the caeca determined by qPCR and immunohistochemistry (IHC). Excretion of the vaccine strain was confirmed prior challenge, with the majority of birds developing antibodies. The new monoxenic vaccine was able to minimize lesions in the caeca demonstrating heterologous protection. No parasites were detected in the liver by IHC in any of the vaccinated birds, compared to non-vaccinated animals. However, in 6 out of 17 birds of the vaccinated group a positive signal was obtained by real time PCR from liver samples with 2 positives being typeable by conventional PCR as genotype 2. Overall, H. meleagridis genotype 2 infection was successfully reproduced. Experimental vaccination with a genetically distantly related genotype 1 was able to reduce lesions, supporting protection by a recently developed vaccine candidate as an efficacious prophylactic strategy.

Caecal microbiota composition of experimental inbred MHC-B lines infected with IBV differs according to genetics and vaccination

Interactions between the gut microbiota and the immune system may be involved in vaccine and infection responses. In the present study, we studied the interactions between caecal microbiota composition and parameters describing the immune response in six experimental inbred chicken lines harboring different MHC haplotypes. Animals were challenge-infected with the infectious bronchitis virus (IBV), and half of them were previously vaccinated against this pathogen. We explored to what extent the gut microbiota composition and the genetic line could be related to the immune response, evaluated through flow cytometry. To do so, we characterized the caecal bacterial communities with a 16S rRNA gene amplicon sequencing approach performed one week after the IBV infectious challenge. We observed significant effects of both the vaccination and the genetic line on the microbiota after the challenge infection with IBV, with a lower bacterial richness in vaccinated chickens. We also observed dissimilar caecal community profiles among the different lines, and between the vaccinated and non-vaccinated animals. The effect of vaccination was similar in all the lines, with a reduced abundance of OTU from the Ruminococcacea UCG-014 and Faecalibacterium genera, and an increased abundance of OTU from the Eisenbergiella genus. The main association between the caecal microbiota and the immune phenotypes involved TCRϒδ expression on TCRϒδ+ T cells. This phenotype was negatively associated with OTU from the Escherichia-Shigella genus that were also less abundant in the lines with the highest responses to the vaccine. We proved that the caecal microbiota composition is associated with the IBV vaccine response level in inbred chicken lines, and that the TCRϒδ+ T cells (judged by TCRϒδ expression) may be an important component involved in this interaction, especially with bacteria from the Escherichia-Shigella genus. We hypothesized that bacteria from the Escherichia-Shigella genus increased the systemic level of bacterial lipid antigens, which subsequently mitigated poultry γδ T cells.

Efficacy of an inactivated Senecavirus A vaccine in weaned pigs and mature sows

Senecavirus A (SVA), commonly known as Seneca Valley virus (SVV) is a causative agent for vesicular disease in swine. It has been found across the globe including the United States, Brazil, and China. Clinical disease caused by this virus is identical to foot-and-mouth disease virus (FMDV). Since FMDV has the potential to cause severe economic consequences in FMDV-free countries, those countries are on high alert for signs of vesicles in swine and an investigation is performed to rule out the presence of FMDV if observed. In countries where SVA cases have continued to occur, investigations and testing can cause a burden on personnel and resources. The objectives of this study were to test the efficacy of a whole-virus inactivated SVA vaccine against challenge in nursery-aged pigs, mature sows, and to assess the protection of passive maternal immunity generated by immunized dams. Animals were given two doses of the vaccine intramuscularly three weeks apart and challenged intranasally two weeks after the second dose. Non-vaccinated animals challenged with SVA developed clinical signs of disease, replicated virus, and developed a neutralizing antibody response. Vaccinated animals had robust neutralizing titers after two doses; and after challenge, did not develop vesicular disease and had limited rectal shedding. Piglets suckling immunized dams and challenged with SVA at 3–6 days-of-age had neutralizing titers prior to challenge and did not replicate or shed virus. An efficacious vaccine could improve swine welfare and reduce the economic consequences of continued foreign animal disease investigations.

ДИАГНОСТИЧЕСКОЕ ЗНАЧЕНИЕ РЕАКЦИИ НЕПРЯМОЙ ГЕМАГГЛЮТИНАЦИИ (РНГА) В СРАВНЕНИИ С ИФА И ДРУГИМИ СЕРОЛОГИЧЕСКИМИ ТЕСТАМИ ПРИ ДИАГНОСТИКЕ БРУЦЕЛЛЕЗА КРУПНОГО РОГАТОГО СКОТА

The results of comparative diagnostic studies of blood sera from cattle for brucellosis from farms with different epizootic situations, performed in the laboratory of the Caspian zonal NIVI - a branch of the FGBNU "FANC RD" and the State Budgetary Institution "Kropotkin Regional Veterinary Laboratory", are presented. The diagnostic efficiency of the indirect hemagglutination test (IHA) with the use of erythrocyte antigen was studied in comparison with other serological methods, such as enzyme immunoassay (ELISA), agglutination test (RA), complement agglutination test (RSC), rose bengal test (RBP), immunodiffusion reaction with O-PS antigen (RID with O-PS antigen). In order to determine the specificity of serological tests, 52 blood serum samples from non-vaccinated animals from a safe farm were examined for brucellosis in RNGA, ELISA, RA, RSK, RBP and RID with O-PS antigen. All blood serum samples were tested with negative results. The sensitivity of the tested diagnostic tools was determined using 82 blood serum samples from cows immunized with the B. abortus 82 vaccine from a brucellosis-prone farm. The results of the studies showed that the infection in the herd proceeds in an acute form, as evidenced by a significant percentage of positively reacting to brucellosis of animals in the RID and high titers of RNHA, RA and RSK. Thus, a positive RNHA was obtained in 20 cases (24.4%), ELISA with the test system of LLC SPF "Sibbiotest" revealed 31 sick animals (37.8%), RSK - 20 (24.4%), RA and RBP - 17 (20.7%), ELISA test system from Biok - 15 (18.3%), RID with O-PS antigen - 9 (11%). It is important to note that the RNGA additionally identified animals that respond positively to brucellosis: to ELISA "Biok" - 5 (6.1%), RA and RBP - 3 (3.7%), RID - 11 (13.4%). The results of our studies have shown that the RNGA method is one of the most sensitive tests that allows to identify a high percentage of animals with brucellosis in a dysfunctional farm (24.4% of the studied ones), which is superior to the results of other serological methods. The results of comparative serological studies of bovine blood sera in RNHA, ELISA, RA, RSK, RBP and RID with O-PS confirmed the specificity of the tests and the high sensitivity of the RNHA method with brucellosis erythrocyte antigen.

Targeting SARS-CoV-2 receptor-binding domain to cells expressing CD40 improves protection to infection in convalescent macaques

Achieving sufficient worldwide vaccination coverage against SARS-CoV-2 will require additional approaches to currently approved viral vector and mRNA vaccines. Subunit vaccines may have distinct advantages when immunizing vulnerable individuals, children and pregnant women. Here, we present a new generation of subunit vaccines targeting viral antigens to CD40-expressing antigen-presenting cells. We demonstrate that targeting the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein to CD40 (αCD40.RBD) induces significant levels of specific T and B cells, with long-term memory phenotypes, in a humanized mouse model. Additionally, we demonstrate that a single dose of the αCD40.RBD vaccine, injected without adjuvant, is sufficient to boost a rapid increase in neutralizing antibodies in convalescent non-human primates (NHPs) exposed six months previously to SARS-CoV-2. Vaccine-elicited antibodies cross-neutralize different SARS-CoV-2 variants, including D614G, B1.1.7 and to a lesser extent B1.351. Such vaccination significantly improves protection against a new high-dose virulent challenge versus that in non-vaccinated convalescent animals.

Maternally Derived Antibody Levels Influence on Vaccine Protection against PCV2d Challenge.

Simple SummaryThe efficacy of vaccination against type 2 porcine circoviruses is influenced by the level of maternally derived antibodies present at the time of vaccination of the piglets. This aspect was investigated by using two types of vaccines either comprised of a whole virus or its single protein. Both vaccines were able to overcome the impact of the maternal antibodies, however, the immunity, measured by serum antibody levels, developed faster for the whole virus type vaccine. The correlation between the maternally derived antibody levels at the time of vaccination and the challenge viral load showed differences amongst the tested lymph nodes. Additionally, it was confirmed also in this study that group oral fluid samples provide a reliable and relatively easy animal welfare-friendly way of estimating the PCV2 status of groups of pigs.Piglets from a porcine circovirus type 2 (PCV2) stable farm of low and high levels of maternally derived antibodies (MDA) against PCV2 were vaccinated either with a whole virus type or a PCV2 ORF2 antigen-based commercial subunit vaccine at three weeks of age. Two non-vaccinated groups served as low and high MDA positive controls. At four weeks post vaccination, all piglets were challenged with a PCV2d-2 type virus strain and were checked for parameters related to vaccine protection over a four-week observation period. MDA levels evidently impacted the outcome of the PCV2d-2 challenge in non-vaccinated animals, while it did not have a significant effect on vaccine-induced protection levels. The humoral immune response developed faster in the whole virus vaccinates than in the subunit vaccinated pigs in the low MDA groups. Further, high MDA levels elicited a stronger negative effect on the vaccine-induced humoral immune response for the subunit vaccine than for the whole virus vaccine. The group-based oral fluid samples and the group mean viraemia and faecal shedding data correlated well, enabling this simple, and animal welfare-friendly sampling method for the evaluation of the PCV2 viral load status of these nursery piglets.

Avidin-Biotin recombinant nucleoprotein competitive ELISA for the detection of peste des petits ruminants virus antibodies in sheep and goats

The present study describes the development of a truncated recombinant peste des petits ruminants virus (PPRV) nucleoprotein (rPPRV-NPN) and its polyclonal antibodies-based immuno-diagnostic assay, Avidin-Biotin (AB) recombinant nucleoprotein competitive ELISA (ABrC-ELISA) for the detection of PPRV antibodies in the sheep and goats. The PPRV N-terminal immunogenic region (1–266 aa) of nucleoprotein (NPN) coding sequence was amplified and cloned into the pETite vector. The rPPRV-NPN with a molecular weight of ∼ 30 kDa was expressed in E. coli, purified, and characterized by SDS-PAGE and immunoblot using standard PPRV specific sera. The Ni-NTA affinity-purified rPPRV-NPN as coating antigen and its hyperimmune serum as competitive antibodies raised in guinea pigs were evaluated as diagnostic reagents in ABrC-ELISA using the known standard panel of sera. The threshold (cut-off) Percentage Inhibition (PI) value was determined as 45 (mean ± 3 SD) based on the reactivity of the known sheep and goats sera to PPRV antibodies [negative (n = 140) and positive (n = 98)] and the assay had a sensitivity of 97 % (95 % Confidence Interval (CI): 91.3–99.4 %) and specificity of 100 % (95 % CI: 97.4–100 %) with an excellent Area under curve (AUC) of 0.997 (95 % CI: 0.99–1.0). On evaluation of diagnostic performance of the assay using the sheep and goats sera (n = 391) from vaccinated, infected, and non-vaccinated animals, the ABrC-ELISA showed the relative diagnostic sensitivity of 95.88 % (95 % CI: 92.56–98.01 %) & 98.77 % (95 % CI: 96.43–99.74 %) and diagnostic specificity of 97.97 % (95 % CI: 94.19–99.58 %) & 90.54 % (95 % CI: 84.64–94.73 %) against indigenous PPR competitive ELISA kit & IDvet Screen® PPR Competition kit, respectively. The study showed that ABrC-ELISA is rapid, sensitive, and specific and can be a better alternative assay for the detection of the PPRV antibodies in the sera of small ruminants for serosurveillance / seromonitoring of PPR not only at the eradication and post-eradication phases in the disease-controlled endemic countries but also in the PPR non-endemic countries.

Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis

Paratuberculosis is a disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). Vaccination is the most cost-effective control method. However, despite the fact that macrophages are the main target cells for this pathogen, the precise mechanisms behind the response of the macrophage to Map infection and how it is modified by vaccination are yet poorly understood. The aim of this study was to investigate the effect of Silirum® vaccination in the early immune response of caprine monocyte-derived macrophages (CaMØs). Peripheral blood mononuclear cells (PBMCs) were obtained from vaccinated and non-vaccinated goats, cultured in vitro until differentiation to macrophages and infected with Map. After a 24 h incubation, Map viability and DNA were assessed in culture by viable colony count and real time quantitative polymerase chain reaction (qPCR). In addition, Map phagocytosis and expression of IL-10, IL-12, IFN-γ, TNF-α, IL-17A, IL-1β, iNOS, IL-6 and MIP-1β were also evaluated through immunofluorescence labelling and reverse transcriptase qPCR (RT-qPCR), respectively. A significant reduction of Map viability was observed in both supernatants (P < 0.05) and CaMØs (P < 0.001) from the vaccinated group. Similarly, the percentage of infected CaMØs and the number of internalized Map by CaMØs (P < 0.0001) was higher in the vaccinated group. Finally, iNOS (P < 0.01) and IL-10 were significantly up-regulated in CaMØs from vaccinated goats, whereas only MIP-1β was up-regulated in non-vaccinated animals (P < 0.05). These results show that vaccination modifies the immune response of CaMØs, suggesting that the phagocytosis and microbiocidal activity of macrophages against Map is enhanced after vaccination.

Comparative proteomics of Brucella melitensis is a useful toolbox for developing prophylactic interventions in a One-Health context

Brucellosis caused by Brucella melitensis is a zoonosis frequently reported in the Mediterranean and Middle-East regions and responsible for important economic losses and reduced animal welfare. To date, current strategies applied to control or eradicate the disease relies on diagnostic tests that suffer from limited specificity in non-vaccinated animals; while prophylactic measures, when applied, use a live attenuated bacterial strain characterized by residual virulence on adult pregnant animals and difficulties in distinguishing vaccinated from infected animals. To overcome these issues, studies are desired to elucidate the bacterial biology and the pathogenetic mechanisms of both the vaccinal strain and the pathogenic strains. Proteomics has a potential in tackling issues of One-Health concern; here, we employed label-free shotgun proteomics to investigate the protein repertoire of the vaccinal strain B. melitensis Rev.1 and compare it with the proteome of the Brucella melitensis 16 M, a reference strain representative of B. melitensis field strains. Comparative proteomics profiling underlines common and diverging traits between the two strains. Common features suggest the potential biochemical routes responsible for the residual virulence of the vaccinal strain, whilst the diverging traits are suggestive biochemical signatures to be further investigated to provide an optimized diagnostic capable of discriminating the vaccinated from infected animals. The data presented in this study are openly available in PRIDE data repository at https://www.ebi.ac.uk/pride/, reference number PXD022472.

Sero-epidemiology of porcine parvovirus, circovirus, and classical swine fever virus infections in India.

Reproductive problems in swine caused by porcine viruses pose a serious threat to the pig industry in developing countries like India. For evaluating the true extent of porcine infections, a total of 1308 representative sera samples were collected from 92 different pig farms covering 8 North-Eastern states and Punjab state of Northern India during a period of 2 years (2011-2013). Sera samples were tested for the presence of antibodies against porcine parvovirus (PPV), porcine circovirus-2 (PCV-2), and classical swine fever virus (CSFV) using commercial enzyme-linked immunosorbent assay (ELISA) kits. In the North-Eastern states, the seroprevalence of CSFV in non-vaccinated animals was 6.30% and that of PCV2 and PPV was 6.28% and 1.24%, respectively. In Punjab, the seroprevalence of CSFV in non-vaccinated animals was 44.44% and seroprevalence of PCV-2 and PPV was 34.07% and 39.10%, respectively. Detection of antibodies against more than one virus revealed that 4.66% animals had co-infection with PCV-2 and PPV, 1.75% with CSF and PPV, 1.98% with CSF and PCV-2, and 1.75% with all the three viruses. The receiver operator characteristics (ROC) curve analysis depicted that piglet mortality, parvovirus, and CSFV were the most important parameters with an AUC value of 0.997, 0.897, and 0.973, respectively. Incidence of single or co-infection with different viruses showed that the occurrence of single infection was significantly more prevalent than co-infection. This study provides useful information to set up future epidemiologic, flock management, and public animal health policies for the prevention and control of PCV-2, PPV, and CSF in India.

Paratuberculosis vaccination specific and non-specific effects on cattle lifespan

Records of cattle vaccination against paratuberculosis (PTB) have been analyzed to determine whether or not non-specific effect (NSE) on overall mortality similar to that observed in BCG vaccinated humans occurs in animals. The results of a previously reported slaughterhouse study on PTB prevalence were used as a reference on the age incidence of advanced patent (clinical) epidemio-pathogenic forms. In the proper vaccine study, cows in 30 cattle farms in the Basque Country, Spain were followed-up for between 1 and 13 years. Vaccinated groups were composed by 1008 (592 right-censored) animals younger than 3 months treated as calves and by 3761 (3160 right-censored) vaccinated at any older age. Controls were 339 (157 right-censored) and 4592 (2213 right-censored) age matched animals, respectively. Individual last year presence in the annual testing was considered age at culling or death. A survival analysis was carried out according age at vaccination of vaccinated versus non-vaccinated animals. PTB age incidence in the slaughterhouse study was subtracted from the difference between vaccinated and non-vaccinated animals at the same age in order to estimate PTB-specific and non-specific effects. The maximum difference was observed at the 2–3 years interval with a 33.9% mortality reduction in the calf vaccinated group. This corresponded also with the maximum NSE that was 24.5% for a PTB incidence of 9.5%. Overall, vaccination afforded to calves a 26.5% yearly mortality protection, split between 11.1% PTB-specific and 15.4% NSE. These results support a NSE on total mortality associated with PTB vaccination that appeared to persist for up to 6–7 years. This confirms for the first time in an animal field study the innate immune system memory predicted by the recently proposed trained immunity theory. Contrasting the literature, no deleterious effects of killed vaccines on females were observed. Mortality reduction would offset vaccination costs and could improve livestock systems efficiency and potentially reduce antibiotic use.Clinical trial registered with Spanish Agency for Drugs and Sanitary products (AEMPS) as 11/012/ECV.

Humoral immune response to Q fever vaccination of three sheep flocks naturally pre-infected with Coxiella burnetii

Qfever is a zoonotic disease caused by the bacterium Coxiella burnetii; Coxiella-infected ruminants are the main reservoir shedding the pathogen during abortion or parturition through birth products. Germany has a long history of small-scale Q fever epidemics in the human population mostly associated with lambing sheep. Therefore, fast and efficient control measures are essentially required to prevent transmission from infected sheep flocks to humans. In our present study, three sheep flocks were vaccinated with an inactivated C.burnetii phase I vaccine after a field infection with C.burnetii was diagnosed. Serum samples and vaginal swabs were collected at different time points to evaluate the extent of the outbreak and the consequences of the vaccination. The serum samples were examined by phase-specific IgG phase I and phase II ELISAs and a commercial ELISA, simultaneously detecting both phase variations. Moreover, vaginal swabs were analysed by qPCR. The fourth flock with no Q fever history and non-vaccinated animals were used as a control group to evaluate the phase-specific ELISAs. The inactivated C.burnetii phase I vaccine induced an IgG phase II response and boosted the humoral immune reaction against natural pre-infections. Furthermore, the longevity of vaccine-induced antibodies seems to depend on previous infections. Around 16 months after primary vaccination, mainly IgG phase I antibodies were detectable. Vaccination did not prevent shedding at the next lambing season. Most interestingly, the phase-specific ELISAs revealed more C.burnetii positive animals than the blended ELISA-Assay. Taken together, phase-specific ELISAs are suitable tools to provide insights into natural- or vaccine-induced humoral immune responses to C.burnetii in sheep.

The 1B vaccine strain of Chlamydia abortus produces placental pathology indistinguishable from a wild type infection.

Chlamydia abortus is one of the most commonly diagnosed causes of infectious abortion in small ruminants worldwide. Control of the disease (Enzootic Abortion of Ewes or EAE) is achieved using the commercial live, attenuated C. abortus 1B vaccine strain, which can be distinguished from virulent wild-type (wt) strains by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Published studies applying this typing method and whole-genome sequence analyses to cases of EAE in vaccinated and non-vaccinated animals have provided strong evidence that the 1B strain is not attenuated and can infect the placenta causing disease in some ewes. Therefore, the objective of this study was to characterise the lesions found in the placentas of ewes vaccinated with the 1B strain and to compare these to those resulting from a wt infection. A C. abortus-free flock of multiparous adult ewes was vaccinated twice, over three breeding seasons, each before mating, with the commercial C. abortus 1B vaccine strain (Cevac® Chlamydia, Ceva Animal Health Ltd.). In the second lambing season following vaccination, placentas (n = 117) were collected at parturition and analysed by C. abortus-specific real-time quantitative PCR (qPCR). Two placentas, from a single ewe, which gave birth to live twin lambs, were found to be positive by qPCR and viable organisms were recovered and identified as vaccine type (vt) by PCR-RFLP, with no evidence of any wt strain being present. All cotyledons from the vt-infected placentas were analysed by histopathology and immunohistochemistry and compared to those from wt-infected placentas. Both vt-infected placentas showed lesions typical of those found in a wt infection in terms of their severity, distribution, and associated intensity of antigen labelling. These results conclusively demonstrate that the 1B strain can infect the placenta, producing typical EAE placental lesions that are indistinguishable from those found in wt infected animals.

Different clinical presentations of subgenotype 2.1 strain of classical swine fever infection in weaned piglets and adults, and long-term cross-protection conferred by a C-strain vaccine

Classical swine fever is an important swine disease in China, and sporadic outbreaks with mild clinical signs despite compulsory vaccination have raised questions about the virulence and pathogenicity of prevalent subgenotype 2.1 strains, and the ability of C-strain vaccines to cross-protect against them. To investigate this, three field isolates were evaluated in experimentally infected piglets and compared with the highly virulent reference Shimen strain. Clinical signs for the field strains ranged from mild to severe, and mortality ranged from 0 to 80 %. These data show differences in virulence among the subgenotype 2.1 field isolates and support the use of field strain GD191 as a genotype 2 challenge virus to assess efficacy of C-strain vaccines.In contrast to the historical genotype 1 strain, which caused acute infection with significant virus shedding in non-vaccinated animals, the subgenotype 2.1 GD191 strain produced different clinical manifestations in weaned piglets and adults. Adult pigs showed subclinical infection with viral shedding, whereas weaned piglets showed overt signs of infection. Efficacy of, and duration of immunity conferred by a C-strain vaccine were assessed using the reference Shimen strain and field isolate GD191 at 12 and 15 months after vaccination. A robust antibody response and sterilising protection were seen in all vaccinated animals and lasted up to 15 months post-vaccination. This study confirms that C-strain vaccines confer both clinical and virological protection against the historical genotype 1 Shimen strain and cross-protection against the prevalent genotype 2 field strain.

Safety and efficacy of an attenuated heartwater (Ehrlichia ruminantium) vaccine administered by the intramuscular route in cattle, sheep and Angora goats

Heartwater is an economically important tick-borne disease of ruminants in Africa. The current commercial vaccine uses live Ehrlichia ruminantium from blood of infected sheep, requires antibiotic treatment during infection, needs to be administered intravenously and does not protect against all South African isolates. An attenuated tissue culture vaccine not requiring antibiotic treatment and effective against different field strains in small groups of goats and sheep was reported previously. The objective of the present study was to test safety and efficacy of this vaccine administered by intramuscular (i.m.) inoculation in larger groups of sheep, Angora goats and cattle. Animals were vaccinated via intravenous (i.v.) and i.m. routes and received E. ruminantium homologous challenge by feeding of infected ticks or by i.v. inoculation of infected blood. For vaccine titration in sheep and goats, the optimum safe and efficacious dose was determined using 2 ml equivalent of 102–105 culture-derived live elementary bodies (EBs). Similarly, the vaccine was titrated in cattle using 5 ml containing 105–107 EBs. Seventy percent of i.v. vaccinated and 9.7% of i.m. vaccinated Angora goats receiving 105 EBs, developed severe reactions to vaccination and were treated. These treated animals and the remaining 90.3% of i.m.- vaccinated goats showed 100% protection against i.v. or tick challenge. Sheep and Angora goats vaccinated i.m. with 104 EBs had no vaccination reactions and were fully protected against i.v. or tick challenge. Similarly, vaccinated cattle (dose 106 EBs) did not react to vaccine inoculation and were fully protected against i.v. or tick homologous challenge. Control non-vaccinated animals reacted severely to challenge and required oxytetracycline treatment. This successfully demonstrated that Angora goats, sheep and cattle can be safely vaccinated with the attenuated E. ruminantium Welgevonden vaccine via the i.m. route, with no clinical reactions to vaccination and 100% protection against virulent i.v. and homologous tick challenge.

The efficiency of vaccination against cattle necrobacteriosis using an experimental vaccine with a new adjuvant complex

The purpose of the study was to analyze the effectiveness of the adjuvant complex based on saponin, glycerin and chitosan in the experimental series of an inactivated vaccine against cattle necrobacteriosis. 4 groups of animals (30 in each group) were selected for the study, aged 6 to 24 months. The first (1st) group consisted of calves 6 months old, the second (2nd) group consisted of animals 12 months old and the third group – animals 24 months old. The control 4 (fourth) group (non-vaccinated animals) contained 10 animals of each age group – 6, 12 and 24 months old. The animals were vaccinated using a needle-free injector at a dose of 0.4 cm3 in two points at a distance of 7–10 cm (0.2 cm3 each) twice at intervals of 4-6 weeks. Revaccinated after 6 months, with a single dose of 0.4 cm3 (at two points, 0.2 cm2 each). It was found that the vaccine with the multicomponent adjuvant had low viscosity and hardening temperature, moderate reactivity, high stability during storage, prolonged immunity in vaccinated animals. The immunity in animals was detected 21 days after the first vaccine administration and lasted at least 18 months after the last immunization. The necrobacteriosis incidence in animals of the control non-vaccinated group was 70-80%. During double vaccination, the average level of postvaccinal agglutinating antibodies in serum of experimental groups of animals was 312.08±74.12 – 62.13±69, 07. Humoral response in vaccinated animals provided reliable protection against Fusobacterium necrophorum disease within two years (observation period). Absence of disease symptoms in vaccinated animals during 2 years testified to the high preventive efficiency of the vaccine. The results obtained confirm the validity of the application of multicomponent adjuvant in inactivated sorbed vaccine as a means of specific prevention of animal necrobacteriosis.

Gilt Vaccination with a Mixed Administration of a PRRS MLV and a PPV1 Subunit Vaccine Protects against Heterologous PRRSV1 Infection and Prevents Detrimental Effects on Piglet Performance.

The efficacy of the combined administration of a porcine reproductive and respiratory syndrome (PRRS) modified live virus (MLV) vaccine and a porcine parvovirus 1 (PPV1) subunit vaccine in gilts was addressed in two experiments. Experiment A aimed to establish a 4-week onset of immunity (OOI). Gilts were randomly distributed in three treatment groups: non-vaccinated control animals (group 1), animals vaccinated with the combined vaccine (group 2), and a third group that consisted of animals vaccinated with the PRRS MLV vaccine alone (group 3). Four weeks after the first vaccination, gilts were challenged with a heterologous PRRS virus 1 (PRRSV1) and euthanized three weeks after. Besides this, experiment B pursued a 17-week duration of immunity (DOI). In this case, gilts were distributed in the same treatment groups, but for the third group, which consisted of non-vaccinated, non-challenged animals were used instead. For the DOI assessment, gilts were artificially inseminated 4 weeks after the first vaccination, challenged at day 90 of gestation, and followed up, together with their offspring, until day 20 post-farrowing. Serology and viremia post-challenge were determined in gilts from both experiments, while farrowing and piglet performance were only evaluated in experiment B. Overall, the combined vaccine helped to protect gilts from viremia post-challenge and, consequently, to prevent PRRS clinical symptoms and diminish the proportion of piglets infected congenitally or early in life. The combined vaccine also elicited a significant improvement in piglet survival rate and growth performance until weaning. The present results reveal efficacy and lack of interference of the mixed use of the tested vaccines against PRRSV1 infection, with at least 4-week OOI and 17-week DOI.