Local tissue reaction to the administration of an inactivated Brucella ovis saline-in-oil vaccine by the intraperitoneal route

Abstract

An inactivated Brucella ovis saline-in-oil vaccine? was administered to 14 adult ewes using both the intraperitoneal route and the subcutaneous route. Pairs of animals were necropsied at intervals between 24 hours and ten weeks after injection. The nature of the local inflammatory reaction to the administration of the vaccine was similar at all sites. The lesion consisted of granulomatous inflammation arranged around droplets of oily vaccine. Diffuse peritonitis was seen at necropsy in 12 of the 14 animals. A local extraperitoneal infammatory response at the injection site was present in four animals despite careful attempts to deposit the vaccine within the abdominal cavity. A second study of 30 rams vaccinated by the intraperitoneal technique confirmed that extraperitoneal deposition of vaccine commonly occurred and that approximately 20% of animals vaccinated by the intraperitoneal method still had peritonitis six months later.     

Vaccination of pigs against Aujeszky's disease by the intradermal route using live attenuated and inactivated virus vaccines.

Inactivated and live Aujeszky's disease virus vaccines were administered intradermally using a special device without a needle. The 88 pigs were vaccinated at the beginning of the fattening period, both under experimental conditions and in commercial herds. All the pigs were challenged at the end of the fattening period in isolation units. The same vaccines were also injected intramuscularly. Vaccination by the intradermal route induced good protection, similar to that conferred with live virus vaccine injected intramuscularly. The inactivated virus vaccine was not as effective when it was injected by the intradermal route. In animals vaccinated intradermally, there were no local lesions in the meat, but very small nodules were found in the dermis; these do not affect carcass quality. The effects of challenge exposure depended on the initial health of the animals, and a synthetic value (delta G) was used to interpret the data. In fattening pigs, intradermal vaccination required less animal constraint than intramuscular injection; administration could be verified by the presence of a papule at the site of inoculation, and pigs could be vaccinated while they were feeding. Injection without a needle also helps avoid bacterial contamination under farm conditions.     

Antibody Response in Goats Vaccinated with Liposome-adjuvanted Clostridium perfringens Type D Epsilon Toxoid

A trial was performed using 20 goats to evaluate the antibody responses to a liposome-adjuvanted Clostridium perfringens epsilon toxoid vaccine (LIPV). The antibody response was compared with those produced by epsilon toxoid vaccines prepared using aluminium hydroxide (ALV) and incomplete Freud's adjuvant (FAV). The animals were allocated to four groups at the beginning of the trial. The animals in group 1 were vaccinated with ALV, while the animals in group 2 received FAV and those in groups 3 and 4 were vaccinated with LIPV. The animals in groups 1 to 3 received three doses of the corresponding vaccine at intervals of three weeks, while those in group 4 received only 1 dose of vaccine at the beginning of the trial. A blood sample was obtained from all the goats at the beginning of the trial and then weekly for 8 weeks. The samples were analysed for epsilon toxoid antibodies by an indirect ELISA technique. No major clinical abnormalities were observed in the animals after vaccination, with the exception of those that received the FAV, which experienced transient lameness. The highest antibody response was observed in the animals vaccinated with FAV, but they presented moderate to severe inflammatory tissue reactions at the injection site. Moderately high antibody responses were obtained with the ALV, with which only minor local reactions were observed. No significant antibody responses were obtained with the LIPV, nor were local reactions observed.     

Vaccination of pigs against Aujeszky's disease by the intradermal route using live attenuated and inactivated virus vaccines

A study was undertaken of the protection induced by inactivated and live Aujeszky's disease virus vaccines. The vaccines were administered using a special device which, without the use of a needle, delivered the preparation intradermally. The trials were performed on 88 pigs which were vaccinated at the beginning of the fattening period both in experimental conditions and in pig herds. All the pigs were challenged at the end of the fattening period in isolation units. The results obtained were compared with those obtained using the same vaccines injected intramuscularly. It was shown that vaccination via the intradermal route induced good protection in the vaccinated animals and was similar to that conferred by live virus vaccine injected intramuscularly. The results, with the inactivated virus vaccine, were not so good when it was injected via the intradermal route. Studies with intradermal vaccination showed no local lesion or very small nodules strictly localized to the dermis. The results also confirmed that the effects of challenge exposure depended on the health status of animals prior to infection and show the necessity to use a synthetic value (delta G) to interpret the data and mainly to compare the results objectively. In fattening pigs this vaccination procedure is attractive because (i) less animal constraint is needed than would be for intramuscular injections, (ii) injection can be checked by the presence of a visible papula at the site of inoculation and, (iii) pigs can be vaccinated in the ham while they are feeding. Injection without a needle also contributes to avoiding bacterial contamination under practical farm conditions of vaccination.     

Serum and mucosal antibody responses and protection in pigs vaccinated against Mycoplasma hyopneumoniae with vaccines containing a denatured membrane antigen pool and adjuvant

Objective To investigate the protective efficacy of a pool of denatured membrane protein antigens of Mycoplasma hyopneumoniae (J strain) in the molecular size range 70 to 85 kDa (F3 antigen) in combination with adjuvants for pigs challenged with M hyopneumoniae .
Design A vaccine efficacy experiment with assessment of serum and respiratory tract antibody responses.
Procedure F3 antigens were emulsified with five different adjuvants. To groups of three pigs per vaccine, four vaccines were given by intramuscular injection, and two vaccines, including one of those given intramuscularly, were given by intraperitoneal injection.
Results Compared to six unvaccinated pigs, animals vaccinated with F3 antigen displayed significantly reduced pneumonia (54% reduction in mean lung score) following experimental challenge. Analysis of post-vaccination, pre-challenge IgG and IgA ELISA antibody absorbances in serum and respiratory tract washings revealed no correlation with lung score. Six weeks after challenge, pigs previously vaccinated intramuscularly mostly demonstrated greater IgG and IgA responses in respiratory tract washings, and greater IgG serum antibody responses, than those vaccinated by intraperitoneal injection.
Conclusion Pigs vaccinated with M hyopneumoniae antigens in the molecular size range of 70 to 85 kDa showed a significant reduction in lung lesions compared with unvaccinated control animals after experimental challenge. IgG and IgA antibody concentrations in serum and respiratory tract washings after vaccination do not provide a useful prognostic indicator of protection from enzootic pneumonia.     

Impact of field vaccination with a Theileria annulata schizont cell culture vaccine on the epidemiology of tropical theileriosis

Tropical theileriosis, caused by Theileria annulata, is an important tick-borne disease of cattle. A cell culture attenuated vaccine has been developed in our laboratory by long-term in vitro propagation of the schizont stage of the parasite. A longitudinal study was conducted at selected farms housing indigenous, cross-bred and exotic animals to investigate the effect of vaccination on the epidemiology of the disease. A total of 120 animals in 4 age groups were vaccinated with the vaccine before the onset of disease season. An equal number of age-matched animals were kept as controls at the same sites. Animals were monitored for 14 months at monthly intervals. The 97.5% vaccinated animals showed a rise in antibody titres 1 month post-vaccination, as determined by single dilution ELISA. The 78.3% of non-vaccinated animals became sero-positive over the period of observation. Mean antibody titres were significantly higher in vaccinated than non-vaccinated animals. Cross-bred animals showed higher antibody titres followed by exotic and indigenous animals in both the vaccinated and non-vaccinated groups. However, the antibody titres in animals of different ages were similar. The 36.7% vaccinated and 64.2% non-vaccinated animals became carriers (<0.5% piroplasms in erythrocytes) during the observation period. Clinical cases of theileriosis were recorded only in the non-vaccinated group suggesting that vaccinated animals were sufficiently immune to withstand field tick challenge for at least 14 months.     

Protective effects of intranasal vaccination with plasmid encoding pseudorabies virus glycoprotein B in mice

Intranasal administration of plasmid DNA encoding glycoprotein B of pseudorabies virus into mice induced both serum and secretory antibody responses. These mice resisted intranasal challenge with lethal dose of the virus, but did not intraperitoneal challenge. On the other hand, intramuscular injection of the plasmid induced less secretory and higher serum antibody responses than those of intranasally vaccinated mice. None of them was protected from virus challenge. The present results suggest that administration of plasmid DNA encoding glycoprotein B by respiratory mucosal route generates local secretory antibodies which serve to protect animals from pseudorabies virus infection.     

Influence of vaccination route on the efficacy of Aujeszky's disease deletion-mutant vaccine.

In order to compare the effect of the route of immunization on the efficacy of a modified live Aujeszky's disease (AD) vaccine, which had deletions in both thymidine kinase (TK-) and glycoprotein gIII genes (gpIII-), 20 six-week-old pigs were vaccinated by either the intramuscular (IM) (n = 10) or subcutaneous (SC) (n = 10) route. All the animals, including five non-vaccinated control animals, were challenged with virulent AD virus 22 days after vaccination. Four of five non-vaccinated animals died within 12 days after challenge. Although none of vaccinated animals died, three of animals in the SC group exhibited clinical signs, and average daily gains in the SC group were depressed. The animals in the IM group were not found to shed challenge virus, but those in the SC group shed the virus up to 9 days. Virus neutralizing antibody titers in the vaccinated animals were low or non-detectable by 21 days after vaccination. A glycoprotein gII (gpII) screening ELISA detected gpII antibody in all animals in the IM group. While, only 30% of animals in the SC group were positive by the same test. The results of this study indicate that TK-, gpIII modified live AD virus vaccine is effective against challenge with virulent AD virus; however, vaccination by the SC route reduced vaccine efficacy in comparison with IM route.     

Protection of mice against Chlamydophila abortus infection with a bacteriophage-mediated DNA vaccine expressing the major outer membrane protein

A bacteriophage-delivered DNA vaccine against Chlamydophila abortus was constructed by cloning a eukaryotic cassette containing the ompA gene (which expresses the Major Outer Membrane Protein) into a bacteriophage lambda vector. Four groups, each of 20 BALB/c mice were inoculated separately with the phage vaccine, a conventional DNA vaccine based on the same ompA expression cassette, a live attenuated vaccine (strain 1B) or the empty phage vector. The phage and DNA vaccines and empty phage vector were administered intramuscularly on days 0, 14 and 28; the attenuated vaccine was given once on day 0. Half the animals in each group were challenged on day 42 by intraperitoneal injection of live C. abortus and sacrificed on day 49. Phage-vaccinated mice developed moderate antibody levels against C. abortus and yielded higher levels of IFN-γ and IL-2 compared with the attenuated live vaccine group. Clearance of chlamydiae from spleens was significantly better in the attenuated vaccine group compared with the phage vaccine group, while both groups were significantly superior to the DNA vaccine and control groups (p<0.01). Although levels of protection in the mouse model were lower in phage-vaccinated animals, than in 1B vaccinated animals, phage vaccines offer several other advantages, such as easier handling and safety, potentially cheaper production and no chance of reversion to virulence. Although these are preliminary results in a model system, it is possible that with further optimisation immunization with phage vaccines may provide a novel way to improve protection against C. abortus infection and trials in large animals are currently being initiated.     

Failure to stimulate a local immune response in the mammary gland of the sow using intraperitoneal "priming"

An experiment was carried out to determine whether a local immune response could be produced in the mammary gland of sows by "priming" the animals with an intraperitoneal injection of antigen without adjuvant. Adjuvant was not used in the immunization regimen because it had been shown to induce acute peritonitis and multiple abscess formation when injected intraperitoneally in pigs. Four pregnant sows were "primed" by intraperitoneal injection of ferritin; a second dose of the antigen was given two weeks later into the lumen of the jejunum. Another group of pregnant sows was given two intra-muscular injections of ferritin two weeks apart and a third group of sows served as non-immunized controls. Antibody titres and concentrations of IgG, IgA and IgM in colostral and milk whey suggested that a local immune response had not been stimulated in the mammary glands of these sows. Mammary tissue was examined for the presence of immunoglobulin-containing and specific antibody-containing plasma cells. Results of these studies confirmed that intraperitoneal/intrajejunal injection of ferritin without adjuvant was not successful in inducing a local immune response in the mammary gland.     

A live bovine herpesvirus-1 marker vaccine is not shed after intramuscular vaccination

It should be established, whether animals vaccinated intramuscularly (IM) with a live Bovine herpesvirus type 1 (BHV-1) marker vaccine become viremic and/or excrete vaccine virus with nasal discharge. Five cattle, seronegative for BHV-1, were vaccinated with an overdose of the vaccine (Bovilis IBR marker live) via the IM route. Nasal swabs and blood samples were taken at regular intervals and tested for BHV-1 in a virus infectivity assay. In addition, a polymerase chain reaction (PCR) specific for BHV-1 DNA was performed on the blood samples. BHV-1 neutralizing antibody titres were determined in the sera taken prior to the vaccination and four weeks after immunisation. AIl animals were successfully vaccinated as judged by the development of BHV-1 neutralising antibodies. However, all nasal swab samples were tested negative for vaccine virus, and all blood samples were found negative for BHV-1 vaccine virus and BHV-1 specific DNA. From these data it can be concluded that the vaccine virus was not excreted with nasal discharge after IM vaccination and that the vaccinated animals did not have a detectable viremia. Therefore, it is recommended to apply the tested BHV-1 marker live vaccine by the IM route in situations where it is undesirable that the vaccine virus is excreted.     

The serological response of rams to four methods of vaccination against Brucella ovis infection

A comparative study was made of the serological response of rams to an inactivated Brucella ovis saline in-oil vaccine administered either once or twice by either the subcutaneous or the intraperitoneal route. The serological response of rams to two spaced doses of vaccine was more consistent and more persistent than when a single dose of vaccine was used. The rise in titre was more rapid and the final titre of greater magnitude when the subcutaneous route of administration was used in comparison with the intraperitoneal route. On a serological basis, the most satisfactory vaccination technique investigated consisted of two subcutaneous injections of vaccine administered ten weeks apart. The least satisfactory technique was a single intraperitoneal injection.     

The response of vaccinated rams to experimental challenge using Brucella ovis

One hundred and thirty eight rams were allocated to four experimental groups. An inactivated Br. ovis vaccine was administered either once by the intraperitoneal route (1 i/p), twice by the intraperitoneal route (2 i/p), or twice by the subcutaneous route (2 s/c), and the last group was left unvaccinated. They were then challenged by the intravenous inoculation of between 123 and 1.23 x 108 live Br. ovis bacteria. The number of rams that succumbed to infection within the four groups was 4135 (11%) for the 2 s/c, 7133 (21%) for the 2 i/p, 9135 (26%) for the 1 i/p and 18135 (51%) for the unvaccinated rams. Vaccination reduced the number of rams that succumbed to experimental challenge and although there were differences between the vaccinated groups, these were not statistically significant. Following challenge, unvaccinated rams were the first to excrete Br. ovis in their semen three weeks following inoculation. Next, those vaccinated by either one or two doses by the intraperitoneal technique began to excrete the organism (five weeks) and then finally those rams vaccinated twice by the subcutaneous route (seven weeks).     

Safety of a non-adjuvanted therapeutic vaccine for the treatment of feline dermatophytosis

The safety of a non-adjuvanted inactivated fungal vaccine for the treatment of dermatophytosis in cats was investigated in two studies: a controlled laboratory study, and a placebo-controlled double-blind field study with a cross-over design in Europe. In the laboratory study, two groups of 10 cats each were administered an intramuscular twofold overdose, followed by five single 1 ml doses, of either vaccine or control product at 14-day intervals. In the field study, cats were treated with three intramuscular injections of 1 ml vaccine administered at 14-day intervals, as recommended by the manufacturer. A total of 89 cats were enrolled in the field study and divided into two groups to receive either vaccine or placebo for the first three treatments, followed by the opposite product for the final three treatments. The cats enrolled in the two studies were 12 weeks of age or older, as recommended by the manufacturer. All the cats were monitored closely for possible injection site reactions, systemic reactions (including changes in rectal body temperature) and adverse events. The results from both studies showed no significant differences between the vaccinated cats and the control or placebo-treated cats with regard to local or systemic reactions. A few mild to moderate local reactions were noted, but these were evenly distributed between the vaccinated and placebo-treated cats and resolved within a few days. No severe or serious adverse events related to the vaccinations were observed.     

An inactivated vaccine for the control of bluetongue virus serotype 16 infection in sheep in Italy

Because no suitable products are at the moment available to safely control the spread of BTV-16 in Europe, an inactivated vaccine was produced from the reference field isolate of bluetongue virus serotype 16. One group of six sheep was vaccinated subcutaneously with the inactivated vaccine twice, on days 0 and 28, whereas a second group of eight sheep was inoculated with saline solution and used as mock-vaccinated control animals. Seventy-eight days after the first vaccination, all sheep were inoculated subcutaneously with a suspension containing 10(6.3) TCID(50) of a virulent reference BTV-16 isolate. Apart from a transient inflammatory reaction at the injection site, no adverse effects were reported following vaccination. All vaccinated animals developed high titres (7.3-9.3log(2)(ED50%/50 microl)) of virus-specific neutralising antibodies and were resistant to challenge with BTV-16. Conversely, following challenge, control animals developed hyperthermia and long lasting high-titre viraemia.     

Passively transferred immunity to IBD following live vaccination of parent chickens by two different routes

Vaccination of broiler breeder parents with live infectious bursal disease (IBD) vaccine by the oral route resulted in higher and more persistent quantitative gel diffusion precipitins than vaccination by intramuscular injection. All the progeny of the orally vaccinated parents had maternally derived IBD antibody (MDA) at hatching while MDA was detected in only 35 per cent of the progeny of the intramuscularly vaccinated parents. This MDA had disappeared completely at 14 and eight days respectively. Susceptibility to IBD challenge began at eight days in the first group and at one day old in the second. The age of 100 per cent susceptibility occurred at 19 and 11 days in the respective groups.     

Humoral immune response in breeding sows after vaccination with a herd-specific Chlamydophila abortus vaccine

A chlamydial vaccine efficacy trial with assessment of the clinical acceptability and serum antibody responses was performed in breeding sows. A BGM cell culture derived vaccine containing 10(8)/ml formalin-inactivated purified elementary bodies (Eb.) in sterile 0.15 M saline was prepared from Chlamydophila (Ch.) abortus strain OCHL03/99 which has been isolated in the herd from a sample of vaginal discharge. Vaccination was performed as a randomised trial with parallel treatment of a vaccinated group (25 sows) and non-vaccinated control group (20 sows). Sows received two 2.0-ml doses of vaccine intramuscularly at a three week interval. Control sows were dosed with sterile 0.15 M saline, accordingly. Serological response to vaccination was measured by ELISA with a total of 204 blood serum samples (114 from the vaccine group; 90 from the control group) using crude chlamydial LPS as the antigen. Compared to the control group, vaccinated sows showed a marked primary and secondary IgG serum antibody response following the two vaccinations. Antibody levels peaked between week 7 and 14 after priming vaccination, declined incrementally until week 27 but remained significantly higher than the corresponding sham-immune control levels and the prevaccination values of the vaccine group (p < 0.05). Western blot analysis of solubilized whole Eb. of Ch. abortus, Ch. pecorum, and Chlamydia (C.) suis with pre- and postvaccination sera confirmed that vaccination induced an antibody response preferentially against a range of 13 chlamydial antigens including the 40 kDa MOMP of Ch. abortus. Clinical side effects consisting of a transient mild local inflammatory reaction at the site of injection were observed in approx. 30% of vaccinated sows. These results provide the basis for further clinical evaluation of the Ch. abortus vaccine to protect sows from chlamydia-induced reproductive disorders.     

Long-term immunogenicity and protection against Mycoplasma agalactiae induced by an oil adjuvant vaccine in sheep

The long-term protective immunity of an inactivated mineral-oil adjuvanted Mycoplasma agalactiae vaccine was evaluated in sheep. The antigen suspension was emulsified with a mixture of three mineral oils (Montanide ISA-563, Marcol-52, Montane-80 at the ratio of 30%, 63%, and 7%, respectively). Twenty-two animals were divided in 2 groups (A and B) and immunised with two doses of the vaccine (group A, n = 14) or used as unvaccinated control (group B, n = 8). Five months after the second vaccination, seven animals of group A and four animals of group B were challenged by nasal route with M. agalactiae. The remaining seven vaccinated and four control animals were challenged intranasally eight months after vaccination. The vaccine was able to induce a full-protective immunity preventing the clinical signs of contagious agalactia and the infection by M. agalactiae in all groups of animals irrespective of the time of challenge after booster administration.     

Comparison of protection levels against pseudorabies virus infection of transgenic mice expressing a soluble form of porcine nectin-1/HveC and vaccinated mice

We recently generated transgenic mice expressing a soluble form of porcine nectin-1 (PHveCIg) showing remarkable resistance to pseudorabies virus (PRV) infection. Nectin-1, also known as herpesvirus entry mediator C (HveC), is an alphaherpesvirus receptor that binds to virion glycoprotein D (gD). In order to evaluate the level of resistance to PRV infection induced by the expression of PHveCIg in the transgenic mice, the protective effects of vaccinated and transgenic mice were directly compared. Mice were immunized with a live vaccine, through intraperitoneal injection of PRV strain Begonia (an attenuated vaccine strain deleted for gE and thymidine kinase genes) at 4 weeks before challenge. The vaccinated and transgenic mice were challenged with 10LD(50), 20LD(50) or 50LD(50) of PRV strainYS-81 via intranasal route. In the vaccinated mice, no protection was observed in the challenges with 20LD(50) and 50LD(50). Only two out of six vaccinated mice survived in the challenge with 10LD(50). In contrast, four transgenic mouse lines showed significant resistance to PRV infection, although the survival rates varied in the challenge with each viral dose. These results demonstrate clearly the high potential of transgenic strategy in control of pseudorabies.     

Intranasal vaccination against upper respiratory tract disease (U.R.D.) in the cat—II. Results of field studies under enzootic conditions in The Netherlands with a combined vaccine containing live attenuated calici- and herpesvirus

A live vaccine containing attenuated calici- and herpesviruses was tested in field studies in the Netherlands.This vaccine is administered intranasally. Cats may show local transient reactions in the week following vaccination. The experiments were conducted in three types of infected premises: boarding catteries, homes for stray cats and breeding colonies. In the boarding catteries the vaccine was instilled intranasally at least one week before admission. If this was impossible—like it is in homes for stray cats—the animals were vaccinated at the day of admission and then quarantined for at least 24 hr.In the four boarding catteries 290 cats were vaccinated. Local reations were observed in 25 animals (less than 9%). Five cats (less than 2%) showed clinical symptoms of the disease in spite of vaccination. However these cases were all in the same cattery which also housed stray cats.In three homes for stray cats 257 animals were vaccinated. The percentage of local reactions was much higher (49%) due to suppressed infections and secondary bacterial invaders. Clinical disease was seen in 13 cats (5%). However eight of these cats were in one cattery where some of the requirements of the experiment—c.q. quarantine—could not be met.Prevention of the disease in infected breeding colonies was difficult to achieve because infected parent animals are known to shed virulent virus during rehousing and parturition.The following vaccination program was very successful: parent animals are vaccinated intranasally prior to mating and the pregnant queens again on the day of parturition. The kittens are then vaccinated intranasally at an age of 8–10 days old. With this program the percentage of healthy kittens that could be sold was as high as 98%. This compares to 66% in a previous vaccination program where the queens were given an intramuscular vaccine and the kittens received either several intramuscular vaccinations or one intranasal vaccination on the age of six days.The CH vaccine is available on the Dutch market for one year. The results obtained by veterinary practitioners confirm the good results obtained in the field studies.