Evaluating the protective efficacy of a trivalent vaccine containing Akabane virus,Aino virus and Chuzan virus against Schmallenberg virus infection

Schmallenberg virus (SBV), an arthropod borne pathogen, spread rapidly throughout the majority of Europe since 2011. It can cause a febrile disease, milk drop, diarrhea, and fetal malformation in ruminants. SBV, a member of the Simbu serogroup within the genus Orthobunyavirus, is closely related to Akabane virus (AKAV) and Aino virus (AINOV) among others. In the present study, 4 Holstein-Friesian calves were immunized twice four weeks apart with a multivalent, inactivated vaccine against AKAV and AINOV. Another 4 calves were kept as unvaccinated controls. All animals were clinically, serologically and virologically examined before and after challenge infection with SBV. AKAV- and AINOV-specific neutralizing antibodies were detected one week before challenge infection, while SBV-specific antibodies were detectable only thereafter. SBV genome was detected in all vaccinated animals and 3 out of 4 controls in serum samples taken after challenge infection. In conclusion, the investigated vaccine was not able to prevent an SBV-infection. Thus, vaccines for other related Simbu serogroup viruses can not substitute SBV-specific vaccines as an instrument for disease control.     

Treatment of ovine foot-rot by vaccination with the specific aetiological agent Bacteroides nodosus

Accelerated healing of foot-rot cases after vaccination with Bacteroides nodosus confirms the aetiological role of this organism in the disease. There is a variation in the degree of response. It may range from an increased body weight of affected vaccinated sheep to complete resolution of clinical signs and lesions.The degree of response following vaccination is influenced by the adjuvant incorporated in vaccines. Oil emulsion vaccines were superior to aqueous alum-precipitated preparations. A saponin derivative, Quil A, enhanced the effect of alum-precipitated vaccines in a therapeutic experiment.High agglutinin titres to vaccine strains develop in immunised sheep. Recovery from infection has been demonstrated in animals with high agglutinin titres to the immunising B. nodosus but with low titres to the strain of B. nodosus with which sheep were infected. Antibodies other than those directed against the pili of B. nodosus may be involved in the mediation of the demonstrated therapeutic response.     

Description of the first Schmallenberg disease outbreak in Spain and subsequent virus spreading in domestic ruminants

Schmallenberg disease (SBD) is an emerging disease transmitted mainly among ruminant species by biting midges of the genus Culicoides. Since the Schmallenberg virus (SBV) was first identified in Germany in late 2011, it rapidly spread to other European countries. The aims of the present study were to describe the first SBD outbreak in Spain and to assess the spread and risk factors associated with SBV infection in domestic ruminants from nearby farms during the following year. In March 2012, one malformed stillborn lamb from a sheep farm located in Cordoba province (Southern Spain) was subjected to necropsy. Pathological compatible lesions and molecular analyses confirmed the first SBV infection in Spain. Afterwards, serum samples from 505 extensively reared domestic ruminants from 29 farms were analysed using both blocking ELISA and virus neutralization test against SBV. The overall seroprevalence was 54.4% (CI_95%: 50.0–58.7). Antibodies were detected in 70.6%, 46.0% and 34.8% of cattle, sheep and goats, respectively. A generalized estimating equation model indicated that the main risk factors associated with SBV infection were: species (cattle), age (adult), and absence of animal insecticide treatment. Pathological and molecular results confirmed the presence of SBV in Spain few months after it was firstly identified in Germany. The seroprevalence detected indicates a widespread circulation of SBV in nearby domestic ruminant farms one year after this first outbreak was reported in Spain. Further studies are warranted to determine the spatio-temporal trend of SBV in domestic ruminants in this country.     

Evidence of excretion of Schmallenberg virus in bull semen

Schmallenberg virus (SBV) is a novel orthobunyavirus, discovered in Germany in late 2011. It mainly infects cattle, sheep and goats and could lead to congenital infection, causing abortion and fetal abnormalities. SBV is transmitted by biting midges from the Culicoides genus and there is no evidence that natural infection occurs directly between ruminants. Here, we could detect SBV RNA in infected bull semen using qRT-PCR (three bulls out of seven tested positive; 29 positive semen batches out of 136). We also found that highly positive semen batches from SBV infected bulls can provoke an acute infection in IFNAR^-/- mice, suggesting the potential presence of infectious virus in the semen of SBV infected bulls.     

施马伦贝格病毒

施马伦贝格病毒病(Schmallenberg virus,SBV)是一种新发现的动物传染病,因于2011年底在德国施马伦贝格镇首次发现而临时得名,随后蔓延于西欧(包括比利时、法国、德国、荷兰、意大利、卢森堡、西班牙、英国和丹麦),并分别在奥地利、波兰、瑞典和芬兰等国的牛、山羊、绵羊中检测到抗体。遗传分析显示该病毒与布尼亚病毒科(Bunyaviridae)正布尼亚病毒属(Orthobunyavirus)西姆布血清群病毒(Simbu serogroup viruses)的亲缘关系最密切,西姆布血清群病毒是已知的反刍动物病原,可通过节肢动物媒介(蚊、蠓)传播。施马伦贝格病毒病有2种不同的临床症状:成年牛出现短暂轻微/温和的病症(产奶量减少、发热、腹泻)和新生哺乳动物(牛、羊)死产和先天缺陷。因为同群类似的病毒不是人畜共患病病原,也无该病毒致人发病的证据,但现阶段尚不能完全排除。尽管目前没有特效的药物和疫苗,但因已有类似病毒(赤羽病)的疫苗,疫苗接种应是控制该病的可能选项。因施马伦贝格病毒是一种新发现的病毒,许多方面尚不清楚,还有待于进一步研究。     

Inferences about the transmission of Schmallenberg virus within and between farms

In the summer of 2011 Schmallenberg virus (SBV), a Culicoides-borne orthobunyavirus, emerged in Germany and The Netherlands and subsequently spread across much of Europe. To draw inferences about the transmission of SBV we have developed two models to describe its spread within and between farms. The within-farm model was fitted to seroprevalence data for cattle and sheep farms in Belgium and The Netherlands, with parameters estimated using approximate Bayesian computation. Despite the short duration of viraemia in cattle and sheep (mean of 3–4 days) the within-farm seroprevalence can reach high levels (mean within-herd seroprevalence >80%), largely because the probability of transmission from host to vector is high (14%) and SBV is able to replicate quickly (0.03 per day-degree) and at relatively low temperatures (threshold for replication: 12.3 °C). Parameter estimates from the within-farm model were then used in a separate between-farm model to simulate the regional spread of SBV. This showed that the rapid spread of SBV at a regional level is primarily a consequence of the high probability of transmission from host to vector and the temperature requirements for virus replication. Our results, obtained for a region of the UK in a typical year with regard to animal movements, indicate that there is no need to invoke additional transmission mechanisms to explain the observed patterns of rapid spread of SBV in Europe. Moreover, the imposition of movement restrictions, even a total movement ban, has little effect on the spread of SBV at this scale.     

Ovine campylobacterosis: preliminary studies of the efficacy of the in vitro serum bactericidal test as an assay for the potency of Campylobacter (Vibrio) fetus subsp intestinalis bacterins.

An in vitro serum bactericidal test was developed to assess the efficacy of Campylobacter fetus bacterins. Four experimental monovalent bacterins (either serotype C or A-2) and 2 commercial bivalent bacterins (a "suspect" and an "efficacious" bacterin) with aluminum hydroxide adjuvant were administered to sheep and rabbits from which antiserums were then prepared. The different vaccines were evaluated by comparing the in vitro bactericidal activity of the sheep and rabbit antiserums. Results of the in vitro tests were compared to the protection induced in vaccinated ewes which were orally exposed to C fetus. The sheep and the rabbit antiserums after they were heated at 56 C for 30 minutes were unable to exert a killing effect on C fetus cells. Addition of a fresh homologous complement source to the heated antiserums was necessary to demonstrate the in vitro bactericidal capacity. In the comparison of the suspect and the efficacious commercial bacterins, which both reportedly contain serotype C cells, there was a statistically significant difference in bactericidal activities for serotype C cells of antiserums from sheep 14 days after the 2nd vaccination. There was a corresponding significant difference in the antiserums from rabbits 14 days after the 2nd vaccination. Proportionally, more abortions and stillbirths were observed in the ewes vaccinated with the suspect bacterin and then orally exposed to C fetus-serotype C cells than in those vaccinated with the efficacious bacterin. The results indicated that the ability of vaccinated sheep to overcome infection is reflected in the in vitro bactericidal capacity of antiserum from the animal. Since 89% of the variation in sheep antiserums from 14 days after 2nd vaccination can be accounted for by rabbit antiserums from 14 days after 2nd vaccination, the in vitro bactericidal capacity of rabbit antiserums probably provides a reliable index of the protective effect of bacterins containing serotype C for ewes exposed to the homologous serotype.     

Immunisation against ovine caseous lymphadenitis: efficacy of monocomponent Corynebacterium pseudotuberculosis toxoid vaccine and combined clostridial-corynebacterial vaccines

Sheep were immunised with Corynebacterium pseudotuberculosis toxoid formulated as a monocomponent vaccine with aluminium adjuvant or in combination with 5 clostridial antigens, and also in the combined form with sodium selenate. Immunised and control sheep were experimentally infected 16 days after vaccination and slaughtered and inspected after a further 3 months to determine their resistance to infection. All 3 vaccines afforded an equal and high level of protection; 91% of vaccinated sheep exhibiting no lesions of caseous lymphadenitis compared with 51.5% affected sheep in the control group. Average lesion counts were 1.2 per affected vaccinated sheep and 4.5 per affected control sheep. Antitoxin responses to the clostridial toxoids incorporated in the combined vaccines were not affected by inclusion of the C pseudotuberculosis toxoid or the sodium selenate.     

A comparison of DNA vaccines expressing the 45W, 18k and 16k host-protective antigens of Taenia ovis in mice and sheep

The immunogenicity of DNA vaccines encoding three different Taenia ovis host-protective antigens was compared in mice and sheep. DNA vaccines encoding the 45W, 18k and 16k antigens of T. ovis were constructed. The ability of DNA vaccines encoding the 45W and 18k genes to express antigen was confirmed by Western blotting of transfected Cos-7 cells. BALB/c mice were vaccinated intramuscularly with 45W, 18k or 16k DNA vaccines and the humoral immune response analysed by ELISA. DNA vaccines expressing 45W, 18k or 16k antigen were immunogenic in mice and generated significant titres of antigen-specific antibody. Intramuscular vaccination of outbred sheep with the T. ovis DNA vaccines generated significantly lower titres of 45W-specific antibody and failed to generate 18k or 16k-specific antibody. The findings of this study show that each of the three T. ovis host-protective antigens are amenable to delivery via DNA vaccines, and that the parameters governing the efficacy of DNA vaccines in sheep require further investigation.     

Prevalence of caseous lymphadenitis and usage of caseous lymphadenitis vaccines in sheep flocks

OBJECTIVE: To estimate the prevalence of caseous lymphadenitis (CLA), determine the current usage of vaccines against CLA and to measure the effectiveness of these vaccines on sheep farms. DESIGN AND POPULATION: A survey was undertaken on 223 sheep flocks in New South Wales, Victoria and Western Australia. METHOD: The prevalence of CLA was measured by conventional inspection techniques at abattoirs in lines of sheep that could be traced back to a farm. Managers of the flocks were sent a questionnaire about their vaccine practices, management practices and knowledge of CLA. RESULTS: The average prevalence of CLA in adult sheep in these flocks was 26% and varied from 20% in Western Australia to 29% in New South Wales. About 43% of sheep producers used CLA vaccines; only 12% used them as recommended. Awareness of CLA was highest in Western Australia. More producers would use CLA vaccine if they knew the prevalence of CLA in their flock and producers obtained most information about CLA from vaccine resellers. CONCLUSIONS: Only 10 to 15% of producers are currently achieving effective CLA control through the use of recommended CLA vaccination programs. In Western Australian flocks more than 25% of effectively vaccinated ewes will be sent to abattoirs in the 2 to 3 years after this study. However, large decreases in the prevalence of CLA can be achieved by about 70% of producers by either making adjustments to their vaccination programs or buying a vaccine with a CLA component. Two or three key facts on effective CLA vaccination could be made available at the point of sale of vaccines and from abattoirs that reported the prevalence of CLA to farmers.     

Association of the time that elapsed from last vaccination with protective effectiveness against foot-and-mouth disease in small ruminants

Routine and emergency vaccination of small ruminants against foot-and-mouth disease (FMD) is mandatory in many endemic countries, yet data on the field effectiveness of the vaccines used is scarce. We conducted an investigation of a serotype O FMD outbreak that took place in a sheep and goat pen, and estimated the effectiveness of various routine vaccination statuses. We also evaluated the protection provided by colostrum administration and emergency vaccination. Animals which were routinely vaccinated twice were not clinically affected while disease incidence was observed among animals routinely vaccinated only once (p = 0.004 according to a two-sided Fisher''s exact test). In groups vaccinated only once, there was a significant association between the average time that elapsed since last vaccination and the disease incidence (n = 5; Spearman correlation coefficient: r_s = 1.0, p < 0.01). In addition, non-vaccinated lambs fed colostrum from dams vaccinated more than 2 months before parturition had a mortality rate of 33%. Administration of emergency vaccination 2 days after the occurrence of the index case was the probable reason for the rapid blocking of the FMD spread within 6 days from its onset in the pen.     

The effect of time between doses on serological response to a recombinant multivalent pilus vaccine against footrot in sheep

SUMMARY The response of sheep to a recombinant multivalent footrot vaccine containing pilus antigens was examined after the administration of two doses of vaccine at Intervals ranging from 2 to 52 weeks. Agglutinating antibody titres were measured 3 weeks after the second vaccination and showed that lengthening of the interdose interval results in higher agglutinin titres. The capability of sheep to mount an increasingly strong immune response as the interval between doses is increased provides an opportunity to maximise the usefulness of vaccination by administering the first dose well before an expected footrot transmission period. This advantage of increasing the interdose interval has not been reported for traditional, whole-cell footrot vaccines, and use of the new pilus vaccine in this manner may improve prospects for disease control. Furthermore, sheep given a third dose either 6 or 12 months after their initial two-dose vaccination program achieved significantly higher titres than those elicited after the second dose, suggesting the likelihood of further improvement in disease control in successive seasons.     

Cross-protection from Bacteroides nodosus vaccines and the interaction of pili and adjuvants

The effects of vaccination of Merino sheep with the purified pili or the whole cells of Bacteroides nodosus strain 198, either in oil or alum-oil adjuvant, on the severity of foot-rot induced with the homologous strain (198) and a heterologous strain (217) were determined in a field experiment, on flood irrigated pasture. The efficacy of the whole cell vaccines was comparable to that of purified pili vaccines, against homologous challenge, when both had a similar content of pilus antigen although the purified pili vaccines induced significantly greater homologous pilus agglutinating antibody titres than the whole cell vaccines. However, against heterologous challenge, the whole cell vaccines in oil (CO) or alum-oil (CAO) provided significantly greater protection than a purified pili-in-oil (PPO) vaccine, the number of severely affected feet in sheep vaccinated with PPO being similar to that of the unvaccinated group. The group vaccinated with purified pili in alum-oil (PPAO) was intermediate between these two extremes. The superior performance of the PPAO in comparison to the PPO vaccine, against heterologous challenge, was associated with significantly higher mean ELISA titres to the outer membrane complex. Western blot analyses implicated a role in cross-protection for outer membrane proteins, in particular a protein Mr 78,000. The PPO vaccine produced fewer, smaller and less persistent vaccination reactions at the inoculation sites than did the other vaccines. Bodyweight gains in the period prior to challenge were much lower for the groups vaccinated with CO and CAO than for the controls and those vaccinated with purified pili, due presumably to the larger vaccination reactions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunization of cattle against Aujeszky's disease with inactivated adjuvant vaccine

The experiments with sheep and young cattle were carried out to test the immunizing efficacy of inactivated adjuvant vaccine against Aujeszky's disease. The vaccine application at doses of 1 ml and 2 ml to lambs at the age of eight to ten months caused the neutralizing antibody production with a significant rise of titres after revaccination. A survival of infection induced with a dose of 10(5.5) TKID50 of virulent virus was recorded in 62.5% of once vaccinated animals and in 87.5% of twice vaccinated animals. When applying different doses of vaccines (from 1 to 10 ml) to young cattle, the antibody reaction level was directly dependent on the inoculum quantity. The double inoculation of animals with vaccines of 2 ml and 5 ml caused the neutralizing antibody production at titres of 1:35, or 1:46. The animals, immunized with the live or inactivated IBR-vaccine possessing high antibody titres against IBR-virus, reacted upon the vaccination with inactivated Aujeszky's vaccine anamnestically, by early production of antibodies in high titres. Metaphylactic vaccination (2 ml of vaccine) of cattle in herds with an acute course days, however earlier during five days from the revaccination when it was carried out in seven days following the first vaccination.     

Effect of thimerosal concentration on the efficacy of inactivated Newcastle disease oil-emulsion vaccines

Different quantities of the preservative thimerosal in inactivated Newcastle disease oil-emulsion vaccines were tested to determine the influence on the hemagglutination-inhibition (HI) response of broilers. The effect of thimerosal was measured in vaccines that had been stored for 1, 21, and 52 weeks; HI serology was conducted at 2, 4, and 6 weeks after vaccination. Mean HI titers 4 weeks after vaccination decreased at a significant rate (P less than or equal to 0.001) with increasing concentrations of thimerosal. HI titers 4 weeks after vaccination with 1-week-old vaccine were significantly (P less than or equal to 0.05) higher than those after vaccination with 52-week-old vaccine at all thimerosal concentrations tested. Titers were also significantly higher (P less than or equal to 0.05) after vaccination with 1-week-old vaccine than after vaccination with 21-week-old vaccine at all thimerosal concentrations below about 8.25 mg/ml of antigen. Thimerosal at the levels recommended in commercial vaccines does not significantly decrease vaccine efficacy.     

The measurement of three cytokine transcripts in naïve and sensitized ovine peripheral blood mononuclear cells following in vitro stimulation with bluetongue virus serotype-23

Bluetongue (BT), a serious disease of sheep and some wild ruminants, is caused by bluetongue virus (BTV), a member of the family, Reoviridae. The current research thrust for controlling BT is on development of efficient vaccines, necessitating clear understanding of ovine immunology. At present, comparative studies on cytokine gene expression profiles of naïve and BTV-sensitized peripheral blood mononuclear cells (PBMC) in sheep have not been clearly understood. In the present study, PBMC from naïve and BEI-inactivated-saponin-adjuvanted BTV-1 vaccinated sheep were stimulated in vitro with heterologous BTV-23. At various intervals, RT-qPCR was carried out to estimate cytokine (interferon-gamma, interleukin-12 and interleukin-2) mRNA expressions that are linked to cell-mediated immunity. The results showed that PBMC cytokine profiles were relatively increased both temporally and quantitatively in immunized sheep PBMC compared to naïve ones, suggesting that BTV-1 vaccination may prime immune system that can cross-react with BTV-23 antigens.     

Vaccines against Aujeszky's disease: Evaluation of their efficacy under standardized laboratory conditions

Summary

A standardized test was developed to compare the efficacy of Aujeszky's disease virus (ADV) vaccines under laboratory conditions. Per test 3 groups of 6 to 8 sero‐negative pigs were used. The first vaccination was done at 10 weeks of age. One group was vaccinated once, another was vaccinated twice and the 3rd served as control. Pigs were challenge exposed to the virulent NIA‐3 strain of ADV 12 weeks after the first vaccination. Apart from mortality, average periods of growth arrest, fever and virus shedding after challenge were used as parameters to evaluate vaccine efficacy.

Two inactivated and 4 attenuated vaccines were tested. Two attenuated vaccine viruses were excreted after vaccination. Despite maximal standardization, a considerable variation still existed between the experiments in mortality and growth arrest periods of control pigs after challenge. However, the controls were always more severely affected than the vaccinated pigs. All vaccines except one were effective in preventing death after challenge, but none conferred complete protection. Most vaccinated pigs still lost weight, developed fever and shed virus after challenge. Revaccination after 3 or 4 weeks had little effect, particularly with the attenuated vaccines. The results of the present study indicate that 2 of the attenuated vaccines conferred the best protection, I attenuated vaccine appeared to be as effective as the 2 inactivated ones, and the 4th attenuated vaccine was least effective.     

An in vivo comparison of the efficacy of the heartwater blood and ground-up tick suspension vaccines in calves

Two groups of calves were respectively immunized with heartwater blood (BV) and ground-up tick suspension (GUTS) vaccine. A third group was left unimmunized as controls. No difference in the immune status conferred could be demonstrated between the 2 vaccines at 6 months and 12 months challenge after vaccination. An index, based on the rectal temperature before and during the reaction, was calculated as an aid in evaluating the data. In practice, the evaluation of heartwater vaccination by challenge is more effective at 12 months than at 6 months after vaccination. No effective difference was demonstrated between the 2 vaccines in their immunizing efficacy.     

Immunisation against ovine caseous lymphadenitis: correlation between Corynebacterium pseudotuberculosis toxoid content and protective efficacy in combined clostridial-corynebacterial vaccines

Groups of sheep were dosed with vaccines containing Corynebacterium pseudotuberculosis toxoid combined in varying amounts with 5 clostridial antigens. Resistance of the sheep to infection with C pseudotuberculosis was tested at 1, 6 and 12 months after vaccination by infection with pus from ovine lymph glands actively infected with C pseudotuberculosis. The outcome was assessed 3 months after challenge by slaughter and inspection of the sheep for caseous lymphadenitis lesions. Protection was demonstrated by a significant reduction in the proportion of immunised sheep exhibiting lesions compared with control sheep, and by fewer abscesses in affected immunised sheep than in affected control sheep. A positive correlation was found between amount of C pseudotuberculosis toxoid administered and degree of protection obtained. Chromatographically-purified toxoid induced essentially the same protection, suggesting that anti-toxic immunity is the major factor in protection.     

Immune responses of sheep to louping-ill virus vaccine

The immune responses of sheep to single and double doses of commercially available louping-ill virus vaccine were examined. The susceptibility to challenge of sheep which had been vaccinated but showed a poor response was also investigated. Two injections of vaccine were required to provoke an adequate antibody response and maximum titres were obtained when there was an interval of two to eight weeks between injections. After challenge, viraemia could not be detected in animals with an antibody titre of 20 although increase in the concentration of humoral antibodies indicated that infection had occurred. Vaccinated but seronegative sheep and vaccinated animals with an antibody titre of 10 were also clinically resistant to the challenge, although circulation of virus was demonstrated. That vaccination had sensitised those animals to viral antigen was evident from the reduced viraemias, the early rise in humoral antibody titres and subsequent protection afforded compared to unvaccinated control animals. Thus, animals with minimal antibody titres after vaccination are protected, but it is recommended that vaccines eliciting the highest possible antibody responses will be the most useful under field conditions.