Determination of duration of immunity of calves vaccinated with the Theileria annulata schizont cell culture vaccine

Bovine tropical theileriosis caused by Theileria annulata is a serious haemoprotozoan disease of cattle affecting exotic cattle, their crossbreeds and young indigenous calves. Cell culture vaccines have been developed and used effectively in various countries for the control of this disease. However, the duration of immunity provided by these vaccines is poorly understood. The present experiments were planned to study the duration of immunity in animals after vaccination with the T. annulata (Hisar) schizont cell culture vaccine. Two groups of calves were vaccinated and challenged after a period of 3 and 6 months, respectively. There was no fever in any of the vaccinated calves after challenge. However, the vaccinated animals exhibited mild to moderate enlargement of lymph nodes and parasitological reactions. The parasitological reactions were very mild in calves challenged after 3 months and moderate in calves challenged after 6 months. There was a mild but significant decrease in the haematological values of calves after challenge. A significant rise in the anti-theilerial antibody titres was observed in all calves after vaccination, which increased further, by many folds after challenge. On the other hand, all the challenge control calves showed symptoms of acute theileriosis and died. The observations suggested that the T. annulata (Hisar) schizont cell culture vaccine provided immunity in vaccinated animals for at least 6 months in the absence of field tick challenge. However, there was some decline in immunity after 6 months, if the animals are not exposed to ticks during this period.     

Effect of Vaccination in the Field with the Theileria annulata (Hisar) Cell Culture Vaccine on Young Calves Born during the Winter Season

The responses were monitored of young crossbred calves vaccinated against tropical theileriosis during the winter against a field tick challenge in the disease season. Thirty-eight calves below 2 months of age, born after the end of the disease season, were selected at an organized farm. Twenty-five animals were vaccinated with Theileria annulata (Hisar) cell culture vaccine (developed at CCS HAU Hisar laboratory) after the end of the disease season and 13 calves were kept as non-vaccinated controls. These calves were observed for their susceptibility to theileriosis in the new disease season. There was an increase in antibody titre in 18 of the 25 vaccinated animals one month after vaccination. The antibody titre then declined gradually, but remained higher than those of the non-vaccinated animals at month 0. No fever or other clinical signs of tropical theileriosis were observed in any of the vaccinated animals. Nine out of 25 (36%) vaccinated calves showed occasional piroplasms (<;0.5%) in blood smears. All the vaccinated animals withstood the field tick challenge. On the other hand, 9 of the 13 (69%) unvaccinated calves exhibited occasional piroplasms, and included three clinical cases of tropical theileriosis. These observations suggest that young crossbred calves vaccinated with the T. annulata (Hisar) cell culture vaccine at the end of the disease season were relatively resistant during the next disease season.     

Prevention of bovine respiratory syncytial virus infection and clinical disease by vaccination

A double blind field trial was carried out with a live attenuated bovine respiratory syncytial virus vaccine. The trial involved 530 calves, two to 10 months old, on 27 dairy farms, where respiratory problems due to bovine respiratory syncytial virus infections had been observed during the preceding year. In 17 herds either all calves were vaccinated (nine groups) or all calves received a placebo (eight groups). In 10 herds half the number of calves were vaccinated and the other half kept as non-vaccinated controls. Calves were vaccinated intramuscularly twice with an interval of four to five weeks. These groups were under regular clinical observation and animals were tested periodically for antibodies to bovine respiratory syncytial virus and parainfluenza type 3 virus. Serological examination indicated that no bovine respiratory syncytial virus infection had occurred prior to the first vaccination in August. Vaccination did not cause adverse reactions. Low concentrations of neutralising and complement fixing antibodies were induced by vaccination and a sharp increase of antibody titres was observed after natural infection of vaccinated animals. Infections with bovine respiratory syncytial virus occurred in six out of eight non-vaccinated groups, in nine out of 10 partly vaccinated groups and in only two out of nine completely vaccinated groups. Virus infection in completely vaccinated groups was significantly reduced compared with partly vaccinated and non-vaccinated groups. The incidence of bovine respiratory syncytial virus lower respiratory disease was significantly reduced in completely vaccinated groups compared to non-vaccinated groups. Generally only mild signs of upper respiratory disease were present in completely vaccinated groups after bovine respiratory syncytial virus infection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Investigations on the Influence of Helminth Parasites on Vaccination of Chickens against Newcastle Disease Virus under Village Conditions

Prevalence studies have shown that almost 100% of free-range chickens are infected with a wide range of parasites. The infections are mostly subclinical in nature, resulting in production losses and occasionally mortality. Newcastle disease (ND), on the other hand, results in high mortality rates during epidemics. ND is a limiting factor for increasing poultry production in many tropical countries, where frequent reports indicate vaccination failures. The aim of our study was to investigate the influence of helminths on the antibody response after vaccination against Newcastle disease of free-range chickens naturally infected with parasites. Sixty chickens were divided into six groups, of which three were vaccinated against ND with a live De Soto vaccine, while the other three remained non-vaccinated. One group within the vaccinated groups and the one within the non-vaccinated group was kept naturally infected with helminth parasites, while the other two groups in each set were dewormed with fenbendazole and niclosamide, and one of each of these groups was subsequently infected with Ascaridia galli. After vaccination, all the groups were followed for 5 weeks and their antibody titres were determined weekly using a HI test. All the birds were finally challenged 4 weeks after vaccination with a virulent velogenic ND virus obtained from a field outbreak. All the vaccinated chickens seroconverted and had high antibody levels after 3 weeks, but these dropped to low levels at 4 weeks after vaccination. After challenge, the antibody titres rose in the dewormed groups but not in the parasite-infected groups. After 5 weeks, all the parasite-infected animals had significantly lower antibody titres than the dewormed animals. All the vaccinated chickens survived the challenge infection, emphasizing the importance of the cellular immune response. Further studies are needed to examine the effects of the parasitic infection on protection against ND over a longer period.     

Immunity to Escherichia coli in pigs: Antibody Secretion by the Mammary Gland after Intramammary or Intramuscular Vaccination with an E. coli Vaccine

Indirect hemagglutinating antibody titres in individual gland samples of colostrum and milk from 13 sows were measured. Five of the sows were vaccinated via a mammary gland and five by the intramuscular route with a live formalinised Escherichia coli vaccine and three remained as non-vaccinated controls.

Antibody titres were higher in colostral and milk whey from the vaccinated sows than from non-vaccinated groups. The inoculated gland in the group of sows given vaccine by the intramammary route secreted milk containing markedly more antibodies to the vaccine E. coli strain than did the non-vaccinated glands. Milk from the vaccinated gland did not contain higher titres to heterologous E. coli O antigens than milk from non-vaccinated glands. Serum titres were the same or higher than the titres in colostrum from non-vaccinated glands.

     

Duration of protective immunity and antibody responses in cattle immunised against alcelaphine herpesvirus-1-induced malignant catarrhal fever

ABSTRACT: Protection of cattle from alcelaphine herpesvirus-1 (AlHV-1)-induced malignant catarrhal fever (MCF) has been described previously, using an attenuated virus vaccine in an unlicensed adjuvant. The vaccine was hypothesised to induce a protective barrier of virus-neutralising antibody in the oro-nasal region, supported by the observation of high titre neutralising antibodies in nasal secretions of protected animals. Here we describe further analysis of this vaccine strategy, studying the effectiveness of the vaccine formulated with a licensed adjuvant; the duration of immunity induced; and the virus-specific antibody responses in plasma and nasal secretions. The results presented here show that the attenuated AlHV-1 vaccine in a licensed adjuvant protected cattle from fatal intranasal challenge with pathogenic AlHV-1 at three or six months. In addition, animals protected from MCF had significantly higher initial anti-viral antibody titres than animals that succumbed to disease; and these antibody titres remained relatively stable after challenge, while titres in vaccinated animals with MCF increased significantly prior to the onset of clinical disease. These data support the view that a mucosal barrier of neutralising antibody blocks infection of vaccinated animals and suggests that the magnitude of the initial response may correlate with long-term protection. Interestingly, the high titre virus-neutralising antibody responses seen in animals that succumbed to MCF after vaccination were not protective.     

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.     

Vaccination of cattle against brucellosis using either a reduced dose of strain 19 or one or two doses of 45/20 vaccine

SUMMARY Three groups, each of 14 mature Jersey heifers, were vaccinated. They were mated about 2 months later and those that became pregnant were challenged at about 6.5 months of pregnancy by the conjunctival application of virulent Brucella abortus. Group 1 heifers received 2 doses of B. abortus 45/20 vaccine 2 months apart. Only 5 of the 14 heifers became pregnant, and of these 5 only one resisted challenge. Group 2 heifers received only one dose of 45/20 vaccine, 5 of the 10 challenged resisted infection. Group 3 heifers received 3 × 10⁸ cfu of strain 19. Six of the 10 heifers challenged resisted infection. All of 5 non-vaccinated control cattle became infected. It appeared advantageous to give only one dose of 45/20 rather than 2 as presently recommended. A single dose of 45/20 vaccine induced resistance to virulent B. abortus approximately equal to that given by the reduced dose of strain 19. One dose of 45/20 vaccine stimulated transient serological positivity in 2 of 28 heifers whereas the reduced dose of strain 19 gave rise to persistent titres in 2 of 14 vaccinated heifers.     

LABORATORY AND FIELD STUDIES WITH A BOVINE EPHEMERAL FEVER VACCINE

Bovine ephemeral fever (BEF) virus vaccines, prepared from the brains of suckling mice infected with strain 525 BEF virus, were evaluated in housed cattle and in the field. The virus in lyophilised preparations was stable for 6 months at -50 degrees C. Thirty-four calves, 5 to 18 months old, were used in laboratory vaccination trials. An increase in serum neutralising antibody was detected in 13 of 14 calves initially free of serum antibody, and all 13 failed to develop clinical illness following challenge with virulent BEF virus. Vaccination resulted in no detectable serum antibody increase in 4 calves, 5 months old, with pre-existing antibody of presumed maternal origin. Seven animals, 18 months of age with serum antibody presumed due to previous BEF infection, developed increased antibody titres following vaccination. In 3 animals vaccinated but not challenged, vaccine-induced antibodies decreased to low levels over 5 months. In contrast, the antibody titres following infection with virulent virus in 2 calves were maintained over 5 months. Field trials, involving 236 animals initially free of serum antibody, were conducted on 5 properties near Mackay and 4 properties near Brisbane. Most of 164 animals were vaccinated with a single dose of lyophilised vaccine containing aluminium hydroxide adjuvant. Only 4 animals failed to develop serum antibody and no adverse reactions to vaccination were reported. Natural infection with BEF occurred in 4 herds at Mackay and clinically mild BEF occurred in 3 of 109 vaccinated and 3 of 46 control animals. On the basis of measured serum antibody titres it was assumed that 8 of 53 animals receiving full vaccine volume, 20 of 40 animals receiving half vaccine volume and 18 of 40 control animals became infected with BEF virus. Two dairy herds in Brisbane became naturally infected with virulent BEF virus 7 months after vaccination. Clinical BEF was observed in 8 of 11 control animals and in 3 of 26 animals which received 2 doses of vaccine. Two strains of BEF virus were isolated from unvaccinated animals that developed clinically mild BEF in the field. These strains either failed to infect, or produced subclinical or very mild BEF, when inoculated intravenously into susceptible calves. The anitbody response to natural infection with apparently mild viruses was short-lived, similar to that produced by vaccination.     

A study of antibody levels in wild ruminants vaccinated against rabies

The authors have vaccinated 22 fallow deer (Dama dama) and 10 mouflons (Ovis ammon musimon) against rabies with an inactivated vaccine: 4 fallow deer with 1 ml, 14 fallow deer and 10 mouflons with 2 ml, 4 animals were kept as controls (fallow deer). The antibody responses were checked by fluorescent foci inhibition carried out on blood samples collected during a two-year period. All the animals developed antibody titres and were still protected after 24 months.     

Experiences with a vaccine being developed for the control of swine dysentery

SUMMARY A prototype vaccine that is being developed for the control of swine dysentery (SD) was tested in two groups of experimental pigs. Vaccination induced high circulating antibody titres against the aetiological agent, Serpulina (Treponema) hyodysenteriae .
Pigs in the first trial were vaccinated twice before being challenged orally with the bacteria. Five of 6 unvaccinated animals developed dysentery within a fortnight of challenge, but only 1 of 6 vaccinated pigs showed signs of disease at this time. Unexpectedly, 1 mo after challenge, the surviving unvaccinated pig and 2 remaining healthy vaccinated animals succumbed to the disease. The reason for the development of this late-onset form of dysentery was not clear.
In the second trial, 8 pigs were vaccinated 3 times. Only 2 of these animals (25%) developed severe dysentery after being mixed with infected pigs, whereas 7 of 8 (88%) unvaccinated control pigs in the same pen became diseased. The late-onset form of dysentery was not observed.
The prototype vaccine for SD provided a useful level of protection, and could be used in programs to control the disease in Australia.     

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.     

Viraemia and abortions are not prevented by two commercial equine herpesvirus-1 vaccines after experimental challenge of horses

Eighteen horses, vaccinated on a number of occasions over a period of 12 to 20 months with either a live equine herpesvirus-1 (EHV-1) or an inactivated EHV-1 vaccine, were challenged by the intranasal instillation of the subtype 1 virus isolated from the 1983 outbreak of abortion and paralytic disease at the Lipizzan Stud, Piber, Austria. The prechallenge serum titres of all vaccinated horses were remarkably low, although most horses had received their last vaccine dose only 3 weeks before test-infection. Higher titres were obtained with the inactivated product than with the live virus vaccine. However, no obvious differences were found between the two vaccines in their ability to prevent disease, in that all vaccinated and two 'sentinel' horses became infected and developed viraemia and some degree of clinical disease after challenge; five of the 10 in-foal mares aborted.     

Does multiple oral vaccination of wild boar against classical swine fever (CSF) have a positive influence on the immunity?

We studied the efficacy of multiple vaccinations of wild boar against classical swine fever (CSF) using a C-strain vaccine. The study consisted of two experiments. In the first experiment, 7 to 8 months old animals were vaccinated either three or four times at an interval of 7 days or twice at an interval of 14 or 28 days. In the second experiment, the efficacy of oral immunisation in young boars (3 months old) was examined after fivefold vaccination at intervals of 14 or 28 days. Independently of the immunisation scheme all wild boar developed neutralising antibodies. An evaluation of the antibody titres 28 days after the initial vaccine application showed that single vaccination and triple immunisation at an interval of 7 days induced the highest antibody titres (X > or = 1/80). In multiple vaccinated young boars (vaccinated at intervals of 14 or 28 days) the third vaccination led to a slight reduction or to an only moderate increase of the antibody titre. In a challenge study after the fifth vaccination all wild boar were protected (no viraemia, no virus excretion, no post-mortem virus detection in organs). This was confirmed by the fact that sentinel animals were not affected. Although other immunisation schemes also were effective, booster vaccination at an interval of 28 days is recommended as basic procedure for eradication of CSF in wild boar. Triple vaccination might also be used at the beginning of the control measures.     

Response of Bali cattle (Bos javanicus) to vaccination with Brucella abortus strain 19 in West Timor

A trial was conducted in two villages (one containing cattle infected with brucellosis and one not containing infected cattle) in Timor, Indonesia to determine the serological response to vaccination with Brucella abortus strain 19 in Bali cattle (Bos javanicus) (n = 599). Mature female cattle were immunised with low-dose strain 19 (2x10(8)-6x10(8) colony forming units) and calves (6-12 months) with high-dose strain 19 (4x10(10)-12x10(10) colony forming units). Other mature females and calves were inoculated with sterile vaccine diluent and formed a non-vaccinated in-contact control group. The seroprevalence and mean titres were highest in the vaccinated cattle 3 months after vaccination. These then receded, however, 1% of vaccinated calves and 1.9% of vaccinated cows from the village without infected cattle were still seropositive on the complement-fixation test (CFT) 24 months after vaccination. Non-vaccinated seropositive animals were more likely to have aborted or had a stillbirth and were less likely to have produced a calf than were seronegative cows from the village containing infected animals. We concluded that strain 19 vaccine induced protection in Bali cattle and that this vaccine might play an important role in the control of bovine brucellosis in Timor.     

The effectiveness of foot-and-mouth disease vaccines in Switzerland. I. Screening tests and herd immunity

Protection of the Swiss national cattle herd against foot-and mouth disease is attempted by annual vaccination with inactivated trivalent (O, A, C) vaccines. With the serotype A5 as an example, this paper demonstrates the procedure of potency testing. Serological data obtained with two vaccines in primovaccinated feeder bulls showed that neutralizing antibodies developed within 7 to 14 days post vaccination. There was no statistically significant difference in the anti-serotype antibody titers induced by each of the vaccines; differences were seen between anti-O serotype and anti-A serotype antibody titers, regardless of which vaccine was used. Epidemiological analyses with about 3000 cattle demonstrated that single-vaccinated, and to a lesser degree twice-vaccinated, animals often had short lived immunity; that is, the antibody titers waned within a few months. In contrast, the majority of thrice and multiple vaccinated animals maintained relatively high antibody titers throughout the twelve month period of observation.     

Experimental trials with a thermostable Newcastle disease virus (strain I2) in commercial and village chickens in Tanzania

Antibody responses in indigenous village and commercial chickens vaccinated with 12 thermostable Newcastle disease (ND) vaccine and protection levels against challenge with a virulent field isolate were determined. The antibody response of village chickens vaccinated by eye drop revealed that 30, 60 and 90 days after primary vaccination, the mean log2 HI titres were 6.1, 5.4 and 3.6, respectively, whereas for commercial chickens, the antibody response after 14, 30 and 90 days were 8.2, 5.1 and 4.2, respectively. Village chickens vaccinated orally via drinking water had mean log2 HI titres of 3.4 after 30 days. After booster vaccination, the mean HI titre was 5.4 and 3.3 after 30 and 60 days post-secondary vaccination (i.e. 60 and 90 days after primary vaccination). Antibody response of mean log2 HI titres of 2.6 was recorded 30 days after primary vaccination orally through food; 30 and 60 days after secondary vaccination (i.e. 60 and 90 days after primary vaccination), mean log2 HI titres were 5.3 and 3.2, respectively. All commercial and village chickens vaccinated by eye drop survived the challenge trial whereas village chickens vaccinated through drinking water and food had protection levels of 80% and 60% 30 days after primary vaccination, respectively. However, 30 days after booster vaccination, the protection level was 100%. At 60 days after secondary vaccination, the protection level dropped again to 80% for chickens vaccinated orally. All control chickens used in the challenge trials developed clinical ND and died 3-5 days after inoculation with the virulent virus. Supported by laboratory findings, I2 strain of NDV seemed to be avirulent, immunogenic and highly protective against virulent isolates of NDV. It may be a suitable vaccine to use in village chickens to vaccinate them against ND in rural areas.     

Protective antibody response produced by the chickens vaccinated with green coloured thermostable Newcastle disease virus

The efficacy of green-coloured (GC) I-2 Newcastle disease vaccine was determined in the present study. I-2 vaccine was mixed with a green coloured dye and stored at 4°C for 6 months while assayed for the virus infectivity at a monthly interval. Chickens were vaccinated with the GC vaccine by eye drop. Serum samples were collected from all birds before and after vaccination at weekly interval for 4 weeks and tested for haemagglutination-inhibition (HI) antibody against Newcastle disease virus (NDV). These chickens were challenged with NDV virulent strain four weeks after vaccination. The results showed that there was no difference between the infectivity titres of GC and uncoloured vaccines. However, chickens vaccinated with GC vaccine produced higher HI antibody titres than chickens vaccinated with uncoloured vaccine. Results from the challenge trial showed that all vaccinated chickens survived whereas all unvaccinated chickens died. The findings from this study have shown that the GC vaccine is safe and produced protective antibodies against NDV in vaccinated chickens. Wambura, P. N., 2008. Protective antibody response produced by the chickens vaccinated with green coloured thermostable Newcastle disease virus. Tropical Animal Health and Production.     

Is there a benefit from an early booster vaccination in the control of equine influenza?

Conventional equine influenza vaccination schedules consist of a primary course of two vaccinations given 4-6 weeks apart followed by a third vaccination (booster) given approximately 5 months later. In between the primary course and the third vaccination, horses are generally considered not to be adequately protected against influenza. This study aimed to investigate whether Thoroughbred foals would benefit from a vaccination schedule in which the third vaccination was given earlier than in conventional vaccination schedules. The vaccines used were an inactivated whole virus equine influenza vaccine and an inactivated whole virus combination vaccine containing equine influenza and equine herpesvirus antigens. Four groups of foals were vaccinated with the two vaccines according to a conventional and an accelerated vaccination schedule in which the third vaccination was given 14 weeks after the first administration. In both groups, the fourth vaccination was given at the normally recommended interval of 26 weeks after the third vaccination for the combination vaccine and 52 weeks after the third vaccination with the influenza only vaccine. The horses were 4-11 months of age and seronegative for influenza. Immunological responses after vaccination were monitored for several months using the single radial haemolysis test. The results indicated that 28 weeks after the first vaccination, antibody levels in horses vaccinated according to the accelerated schedule were not significantly higher than in horses vaccinated according to the conventional schedule. In addition, the total level of antibody production (area under the curve) was not significantly different at that point although antibody titres were slightly higher (but not significantly so) between 16-30 weeks in the accelerated schedule. Between the third and fourth doses, horses vaccinated according to the accelerated schedule had antibodies against influenza below the level required for clinical protection for 39 and 18 weeks for the influenza only and the combination vaccine, respectively, whereas those vaccinated according to the conventional schedule had antibody titres below the level for clinical protection for 9-15 weeks in the corresponding period for both vaccines. Horses vaccinated according to the accelerated schedule with the combination vaccine had lower antibody titres after the fourth vaccination than those vaccinated according to the conventional schedule after the third vaccination, although antibody titres prior to vaccination were similar. For the influenza only vaccine, titres after the accelerated fourth administration were not different to those after the conventional third vaccination. There was no benefit from early booster vaccinations with the vaccines used in this study, so for these vaccines the conventional schedule provided better protection than the selected accelerated alternative. This may contrast with some other vaccine formulations, although a direct comparison using similar protocols has not been made.     

Protection against neonatal Escherichia coli diarrhoea in pigs by vaccination of sows with a new vaccine that contains purified enterotoxic E. coli virulence factors F4ac, F4ab, F5 and F6 fimbrial antigens and heat-labile E. coli enterotoxin (LT) toxoid

The efficacy of a new vaccine against neonatal Escherichia coli diarrhoea in piglets containing purified F4ab, F4ac, F5 and F6 fimbriae and detoxified heat-labile toxin (LT) was tested in challenge experiments by the method described by the European Pharmacopoeia (3rd edn, EDQM, Council of Europe, Strasbourg, France). A group of 11 young sows from a herd without E. coli problems was vaccinated 6-8 and 2-4 weeks prior to expected farrowing and another group of nine young sows were non-vaccinated controls. Escherichia coli antibody titres were determined in serum samples taken from the sows before first vaccination and before farrowing and in colostrum samples. The newborn piglets were allowed to suckle colostrum from their mother immediately after birth. The piglets were marked with individually numbered ear tags. Approximately 12 h after birth, 118 piglets from vaccinated sows and 79 piglets from non-vaccinated control sows were challenged by oral instillation of 5 ml of a freshly prepared culture of one of the challenge strains [O8:K87:F4ab (LT+) or O149:K91:F4ac (LT+) or O9:K30:F5 or O9:K103:F6 respectively]. The challenge cultures contained as a mean 6.8x10(9) CFU/ml. After challenge the piglets were observed for 7 days and mortality and morbidity were recorded. Vaccinated sows developed significant levels of antibody titres in colostrum and serum. Control sows stayed at a low/seronegative level. The protective efficacy was excellent because 66.7-87.5% of the piglets from vaccinated sows remained without clinical signs after challenge. Only 0.0-28.0% of the piglets from non-vaccinated sows remained healthy and more than 47.1% of the piglets in this group died after challenge. It is concluded that the new vaccine is very effective in protection of piglets against neonatal E. coli diarrhoea.