Development of post-weaning diarrhoea in piglets. Relation to presence of Escherichia coli strains and rotavirus

Weaning of piglets complicated with an exposure to pathogenic strains of Escherichia coli was scrutinized in two sets. The first set comprised 20 animals representing two litters and the second set included 30 animals from five litters. The piglets were either left as controls or exposed to one or three pathogenic strains of E. coli. Aiming to simulate a natural exposure the challenge strains were spread on the floor of the pens at weaning. In addition the pigs experienced several non-infectious stress factors commonly occurring at that occasion. Some groups were given adrenocorticotropic hormone (ACTH), aiming to simulate a stressful weaning. The balance and the composition of the faecal coliform populations, measured by a metabolic fingerprinting method, was disturbed among all animals following weaning. This disturbance was more pronounced and lasted longer among piglets exposed to pathogenic strains of E. coli. All piglets exposed to pathogenic E. coli shed these strains in faeces. Diarrhoea was induced in the groups exposed to E. coli, but not among the control animals. Pigs not treated with ACTH and subjected to a single pathogenic strain of E. coli became infected but did not develop diarrhoea unless if coinciding with shed of rotavirus. Control pigs excreting rotavirus had no diarrhoea. Diarrhoea was most frequent in the groups exposed to three pathogenic strains of E. coli, and in these groups diarrhoea was seen in the absence of rotavirus. ACTH administration amplified the clinical signs. The litter of origin influenced the development of post-weaning diarrhoea.     

Protection of the nursing pig against experimentally induced enteric colibacillosis by vaccination of dam with fimbrial antigens of E coli (K88, K99 and 987P)

Pregnant gilts were vaccinated with two doses of alhydrogel adsorbed fimbrial antigens of Escherichia coli (K88ab, K88ac, K99 and 987P) supplemented with beta toxoid of Clostridium perfringens type C. Their piglets, and piglets of nonvaccinated gilts, were subsequently orogastrically challenged with one or other of the four fimbrial types of enteropathogenic E coli. Some of the vaccinated animals were reinjected with a single dose of the vaccine during second gestation and their piglets, and piglets of non-vaccinated sows, were challenged the same way as were litters of gilts. Blood serum and colostra were examined for antibodies to the four fimbrial antigens of E coli and for antitoxin to beta toxin of C perfringens type C. It was found that: (1) a highly significant reduction in mortality and morbidity was achieved in vaccinated litters against all four challenge strains of E coli; (2) excretion of K88ab and K88ac but not of K99 and 987P challenge strains was significantly reduced; (3) revaccination of sows by a single dose of the vaccine during second gestation conferred complete protection against mortality and highly significant protection against morbidity; (4) no correlation was noted between colostral or seroagglutinins to fimbrial antigens of E coli and mortality rates in litters challenged with homologous fimbrial types of E coli, but good correlation was found between colostral precipitins to K88 antigens and mortality rates in litters; (5) antitoxin value in 97 per cent of colostrum of vaccinated sows was 10 iu equivalent of C perfringens type C toxin or more per ml of colostrum.     

Prevention of edema disease in pigs by vaccination with verotoxin 2e toxoid.

Pigs in 2 herds with persistent problems with post weaning edema disease caused by infection with verotoxin-2e (VT2e)-producing Escherichia coli O139 were treated with a VT2e-toxoid vaccine. Treatment was performed as a randomized blind field trial with parallel treatment and non-vaccinated control groups. In 1 herd, a group of pigs was injected with adjuvant alone. Pigs were vaccinated at 1 and 3 wk of age and weaned at 4 wk of age. The effect of vaccination was measured by average daily weight gain (ADG), mortality due to edema disease within the 1st 4 wk after weaning, and weight at 3-6 mo of age. Pathological and microbiological examinations were performed on all pigs that died during the 1st 4 wk post weaning. Only pigs from which VT2e+, F18+ E. coli O139 was isolated were categorized as "death due to edema disease." The serological response to vaccination was evaluated by an indirect ELISA. Vaccination had a statistically significant effect on the level of antibodies specific for VT2e in both herds. Vaccination resulted in a statistically significant increase in ADG in the nursery period but not in the grower-finishing period. Vaccination had a statistically significant effect on mortality due to edema disease with an odds ratio of 0.039, indicating that there was almost total elimination of mortality due to the disease in the vaccine groups.     

Verification of peroral vaccination of pigs against enteropathogenic strains of Escherichia coli

In a large herd of pigs where the parenteral immunisation of pregnant sows by polyvalent vaccine against enteral coliinfections of new-born piglets was performed successfully for several years, an increased occurrence of diseases and mortality of sucking piglets was observed. After screening tests in the herd, six strains of E. coli, differing from the strains contained in the commercial vaccine, were isolated. The obtained strains were employed for preparation of peroral vaccine for sows by means of the modified method after Kohler. In two cycles of sows (48 animals in experimental group and 60 animals in control group), this method was compared with the effectiveness of the commonly used commercial polyvalent parenteral vaccine against enteral E. coli infections of new-born piglets (manufactured by Bioveta, Ivanovice in Haná). In experimental sow groups vaccinated perorally by the new vaccine, the number of live-born piglets increased by 0.475 piglet per litter, the number of reared piglets up to the age of 28 days by 0.904 piglet per litter and the mortality till the age of 28 days decreased by 6.1% as compared with the control vaccinated by commercial vaccine. The results were not statistically significant. The advantages and disadvantages of both vaccination methods are discussed.     

The effect of dietary spray-dried porcine plasma on clinical response in weaned piglets challenged with a pathogenic Escherichia coli.

Weaned piglets were used to determine the effect of dietary spray-dried porcine plasma (SDPP) on the clinical response to an infection with a pathogenic Escherichia coli (E. coli) O139:K82 LT(-). The piglets were divided into two groups of 10 animals each. One group was fed the control diet containing soybean(meal) plus whey powder. The test piglets were fed a diet with 8% SDPP. Piglets were orally infected with the challenge strain on days 6 and 7 after weaning. The experimental period lasted 14 days after which the piglets were euthanised and necropsied. Faecal samples were collected daily for bacteriological analysis. Segments of jejunum, caecum and rectum were removed for bacteriological analysis post mortem. Feed intake and weight gain, faecal and condition scores and body temperature were measured daily. In the control and SDPP groups, 6 and 7 piglets died from diarrhoea. The average daily feed intake (ADFI) and average daily gain (ADG) were substantially higher in the SDPP group than in the control group. SDPP-fed piglets generally had a more favourable faecal score and a healthier appearance than did the control piglets. The faecal excretion of E. coli O139:K82 was similar for control and test piglets. There were no diet effects on the E. coli O139:K82 counts at different sites of the intestine. In this experiment, the inclusion of SDPP at an economically acceptable percentage in the diet could not prevent piglet losses due to challenge with a pathogenic E. coli, but improvements of ADG, ADFI and faecal and condition scores were achieved.     

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.     

Development of Post‐weaning Diarrhoea in Piglets. Relation to Presence of Escherichia coli Strains and Rotavirus

Weaning of piglets complicated with an exposure to pathogenic strains of Escherichia coli was scrutinized in two sets. The first set comprised 20 animals representing two litters and the second set included 30 animals from five litters. The piglets were either left as controls or exposed to one or three pathogenic strains of E. coli. Aiming to simulate a natural exposure the challenge strains were spread on the floor of the pens at weaning. In addition the pigs experienced several non‐infectious stress factors commonly occurring at that occasion. Some groups were given adrenocorticotropic hormone (ACTH), aiming to simulate a stressful weaning. The balance and the composition of the faecal coliform populations, measured by a metabolic fingerprinting method, was disturbed among all animals following weaning. This disturbance was more pronounced and lasted longer among piglets exposed to pathogenic strains of E. coli. All piglets exposed to pathogenic E. coli shed these strains in faeces. Diarrhoea was induced in the groups exposed to E. coli, but not among the control animals. Pigs not treated with ACTH and subjected to a single pathogenic strain of E. coli became infected but did not develop diarrhoea unless if coinciding with shed of rotavirus. Control pigs excreting rotavirus had no diarrhoea. Diarrhoea was most frequent in the groups exposed to three pathogenic strains of E. coli, and in these groups diarrhoea was seen in the absence of rotavirus. ACTH administration amplified the clinical signs. The litter of origin influenced the development of post‐weaning diarrhoea.     

对应用遗传工程活菌苗E.coli C_(600)(pMMO85)免疫母猪所产仔猪的攻击试验

应用遗传工程活菌苗E.coli C_(600)(pMMO85),在母猪预产前15～20d,经口服和皮下注射两种途径免疫接种。对免疫母猪所生仔猪,每头攻击E.coli C_(83549)(O149:k91:k(?)8_(?))强毒菌600亿后,再让其吸吮母乳,观察仔猪发生腹泻和排菌情况。结果,对照组3(?)头,在攻击后8～12h均发生腹泻,死亡31头(81.58％),存活仔猪7头,腹泻时间平均2.60d,排菌时间平均5d;而吸吮经两种途径免疫母猪乳汁的仔猪87头,发育正常,仅个别仔猪在攻击1d后发生轻度腹泻,一二天内均能自愈,排菌时间也较对照组大为缩短,平均1.28～1.35d。     

Evaluation of the low dose level of a heat-killed and dried cell preparation of Enterococcus faecalis to prevent porcine edema disease using experimental infection model with enterotoxcemic Escherichia coli in weaning pigs

Porcine edema disease (ED) is caused by Shiga toxin 2e-producing Escherichia coli (STEC). ED has become frequent in pig farms, and the use of antimicrobials has resulted in the development of antimicrobial-resistant STEC. Accordingly, the use of materials other than antimicrobials is requested for the prevention of ED. Oral administration of a heat-killed and dried cell preparation of Enterococcus faecalis strain EC-12 (EC-12) to weaning pigs was previously demonstrated to decrease animal mortality in a STEC-contaminated farm at 0.05% (w/w) dose level. In this study, pigs experimentally infected with STEC were used as a model for ED to evaluate the low dose level of EC-12 to prevent ED. Fifteen 21-day-old pigs were divided into 5 groups: STEC challenge with the basal diet, STEC challenge with EC-12 supplemented at 0.005, 0.01, or 0.05% (w/w) to the basal diet, and no STEC challenge with the basal diet. The challenge was carried out when the animals were 25, 26, and 27 days old using STEC contained in capsules resistant against gastric digestion. All pigs were euthanized at 32 days of age. The daily weight gain, feed conversion ratio, and palpebral edema were improved by supplementation with 0.05% EC-12, but not by the low dose levels. Accordingly, 0.05% level of supplementation was needed for EC-12 to improve clinical symptoms in weaning piglets infected by STEC.     

Enteric-coated pellets of F4 fimbriae for oral vaccination of suckling piglets against enterotoxigenic Escherichia coli infections

To prevent enterotoxigenic Escherichia coli (ETEC) induced postweaning diarrhoea, the piglet needs an active mucosal immunity at the moment of weaning. In the present study, the feasibility of oral vaccination of suckling piglets against F4+ETEC infection with F4 fimbriae was studied. Furthermore, oral vaccination with enteric-coated pellets of F4 fimbriae was compared to vaccination with F4 fimbriae in solution. Therefore, piglets were orally administered 1mg F4 fimbriae in pellets or in solution during three successive days at the age of 7 and 21 days, whereas control piglets were not vaccinated. Five days postweaning (33 days of age), all animals were orally challenged with F4+ETEC. Despite the induction of an immune response upon oral administration of both F4 fimbriae in pellets as in solution, the colonisation of the small intestine by F4+ETEC upon oral challenge could not be prevented. However, a marginal but significant reduction in F4+ E. coli faecal excretion was found in the piglets vaccinated with F4 fimbriae in pellets, indicating that the use of an enteric-coat which protects the F4 fimbriae against inactivation by milk factors and degradation by enzymes and bile improves vaccination.     

Weaning of piglets. Effects of an exposure to a pathogenic strain of Escherichia coli

The influence of weaning on day 32 and a simultaneous challenge with a pathogenic strain of Escherichia coli was studied in eight piglets. Another nine weaned but non-infected piglets were used as controls. The distribution of peripheral blood mononuclear cells (PBMC) into subpopulations, as well as their response when stimulated in vitro by pokeweed mitogen, changed in a similar manner during post-weaning in both groups. In contrast, superior responses were recorded for PBMC collected from the challenged pigs when stimulated in vitro with concanavalin A and with a heat-inactivated extract of the E. coli strain used for infection, respectively. Despite a successful colonization of the challenge strain, no clinical signs of disease were recorded. Nor did the daily weight gain or the number of E. coli, enterococci, or Clostridium perfringens excreted per gram of faeces differ between the groups. However, the weaning induced a marked decrease in the diversity of coliforms in individual piglets, which announced a reduced colonization resistance of that flora. Also, a decreased homogeneity between coliform floras of different piglets was observed following weaning. The decreased homogeneity indicated that different strains of E. coli were predominant in different animals, which may in turn facilitate the spread of pathogenic strains. The enteric changes were more pronounced and lasted longer in infected animals. Still, the influence of a sole pathogenic strain of E. coli was not enough to induce post-weaning diarrhoea.     

Vaccination with type III secreted proteins leads to decreased shedding in calves after experimental infection with Escherichia coli O157

Escherichia coli O157:H7 remains a threat to humans via cattle-derived fecal contamination of food and water. Preharvest intervention strategies represent a means of reducing the pathogen burden before harvest. In this study, the efficacy of a commercially produced type III secreted protein (TTSP) vaccine was evaluated with the use of a commingled experimental calf infection model (30 placebo-treated animals and 30 vaccinates). The calves were vaccinated on days 0, 21, and 42 and infected with 10(9) colony-forming units (CFU) of E. coli O157 by oral-gastric intubation on day 56. Fecal shedding was monitored daily for 14 d. Serologic assessment revealed a robust immune response to vaccination; the serum titers of antibodies against EspA, Tir, and total TTSPs were significantly higher in the vaccinates than in the placebo-treated animals on days 21, 42, 56, and 70. Significantly less (P = 0.011) of the challenge organism was shed by the vaccinates than by the placebo-treated animals on days 3 to 10. Peak shedding occurred in both groups on days 3 to 6; during this period the vaccinates showed a mean log reduction of 1.4 (P = 0.002) and a mitigated fraction of 51%. The number of animals shedding was significantly lower among the vaccinates compared with the placebo group on days 3 to 6 (P ≤ 0.05), with a mean prevented fraction of 21%. No differences in the duration of shedding were observed. Owing to the low challenge shedding in both groups on days 11 to 14 (mean CFU/g < 10; median = 0), no significant differences were observed. These data indicate that TTSP vaccination had protective effects through significant reductions in the number of animals shedding and the number of challenge organisms shed per animal and provides evidence that TTSP vaccination is an effective preharvest intervention strategy against E. coli O157.     

The preventive effect of Bacillus subtilus strain DB9011 against experimental infection with enterotoxcemic Escherichia coli in weaning piglets

Porcine edema disease (ED) is caused by Shiga toxin 2e‐producing Escherichia coli (STEC). Post‐weaned piglets often suffer from ED as a result of intestinal infection with STEC, which causes impaired growth performance and high mortality. Antimicrobial therapy is a curative treatment for piglets infected with STEC, but the emergence of antimicrobial‐resistant STEC has become a serious problem for Japanese pig farmers. Therefore, an alternative strategy other than antimicrobial therapy is needed for the prevention or treatment of ED. In this study, we evaluated the effect of oral administration of Bacillus subtilis DB9011 (DB9011) to prevent the experimental infection of STEC in weaning piglets. Eight 21‐day‐old piglets were divided into two groups: STEC challenge with the basal diet, and STEC challenge with DB9011 supplemented diet. The challenge was carried out when the animals were 25, 26 and 27 days old using STEC contained in capsules resistant against gastric digestion. All pigs were euthanized at 36 days of age. DB9011 improved the symptoms of ED and decreased the number of STEC in the ileal digesta and feces. Accordingly, oral administration of DB9011 in weaned piglets prevents ED through the suppression of the growth of STEC in the ileum.     

Aspects on feed related prophylactic measures aiming to prevent post weaning diarrhoea in pigs

The ability of feed related measures to prevent or reduce post weaning diarrhoea (PWD) was examined in a split litter study including 30 pigs from 6 litters allotted into 5 groups. Four groups were exposed to 3 pathogenic strains of E. coli via the environment at weaning. Three of them were given zinc oxide, lactose+fibres or non-pathogenic strains of E. coil as probiotics. The challenged and the unchallenged control groups were given a standard creep feed. Diarrhoea was observed in all challenged groups but not among uninfected animals, and the incidence of diarrhoea was lower in the group given nonpathogenic E. coli compared to all other challenged groups. The severity of PWD also differed between litters. When corrected for mortality due to PWD, a decreased incidence of diarrhoea was also seen in the groups given zinc oxide or lactose+fibres. The dominating serotype of E. coil within faecal samples varied from day to day, also among diarrhoeic pigs, indicating that diarrhoea was not induced by one single serotype alone. The diversity of the faecal coliform populations decreased in all piglets during the first week post weaning, coinciding with an increased similarity between these populations among pigs in the challenged groups. This indicated an influence of the challenge strains, which ceased during the second week. The group given lactose+fibres was least affected with respect to these parameters. In conclusion feed related measures may alleviate symptoms of PWD.     

液态复合添加剂对断奶1周内仔猪生长性能、养分消化率及血液指标的影响

本试验旨在研究仔猪断奶1周内补饲液态复合饲料添加剂对其生长性能、养分消化率、消化道p H及血液指标的影响。挑选20日龄仔猪105头,断奶前屠宰5头,其他100头在21日龄断奶后分为对照组和试验组,每组5个重复,每个重复10头猪。对照组仔猪饲喂教槽料,试验组仔猪在每次饲喂教槽料后补喂液态复合饲料添加剂(主要成分为酸化剂、中药提取物、壳寡糖、复合离子调节剂)。试验从断奶当天至断奶后第8天,共计7 d。结果表明:试验组的平均日增重高于对照组(P0.05),腹泻率显著低于对照组(P0.05);试验组粗蛋白和钙的表观消化率较对照组显著增加(P0.05);试验组仔猪断奶后第2天和第8天胃部p H、断奶后第8天十二指肠p H、断奶后第4天回肠p H均显著低于对照组(P0.05),断奶后第4天氯离子含量、断奶后第2天和第8天球蛋白、断奶后第4天和第8天血清Ig G和Ig A含量均显著高于对照组(P0.05),而断奶后第2天血清尿素氮显著低于对照组(P0.05)。由此表明,断奶1周内给仔猪补饲液态复合饲料添加剂,可降低消化道p H,提高养分消化率,并改善机体免疫功能,从而提高仔猪断奶1周内的生长性能,降低仔猪腹泻率。     

仔猪大肠杆菌病防治

致病性大肠杆菌是引起仔猪黄痢、仔猪白痢、仔猪水肿病的病原,感染后发病仔猪临床症状以腹泻、水肿等为特征,传染性强,对仔猪生长发育造成影响,甚至导致仔猪急性死亡。通过加强饲养管理、做好清洁卫生及消毒可减少该病发生,还可通过大肠杆菌疫苗免疫怀孕母猪进行预防。该文论述仔猪大肠杆菌病的致病机理、临床症状及防治措施。     

Active oral immunization of suckling piglets to prevent colonization after weaning by enterotoxigenic Escherichia coli with fimbriae F18

Immunoprophylaxis of porcine oedema disease and post-weaning diarrhoea caused by strains of Escherichia coli expressing fimbriae F18 is an unsolved problem. The study was designed to examine whether vaccination with a live F18ac vaccine of unweaned pigs born to sows with F18ac antibody in the colostrum requires preformed fimbriae in the vaccine, and whether protection against the heterologous fimbrial variant F18ab is induced as well. Genetically susceptible pigs were vaccinated orally on three consecutive days, beginning 10 days before weaning with 10(11) CFU of an F18ac culture. Challenge with a dose of 10(7) CFU of E. coli F18 on three consecutive days was initiated 9 or 11 days after weaning. Eighteen pigs given the fimbriated F18ac vaccine and challenged with a strain of the homologous fimbrial variant were protected against colonization; mean faecal viable counts of the challenge strain were >3 log10 lower than those from the 17 non-vaccinated control pigs. The vaccinated pigs developed a significant rise of F18ac IgA serum antibodies. The 23 pigs which had received the non-fimbriated vaccine showed no significant protection and exhibited much lower serum F18ac IgA ELISA reactivities. Eighteen pigs vaccinated with the fimbriated F18ac and challenged with an F18ab strain had faecal viable counts nearly as high as those from 16 non-vaccinated control pigs. It is concluded that only oral vaccines having preformed fimbriae induce protection limited to the homologous fimbrial variant.     

The Effect of Season and Vaccination for Glässer's Disease and Post‐weaning Colibacillosis in an Outdoor Pig Unit Endemically Infected with Virulent Strain of Haemophilus Parasuis Serotype 5 and Pathogenic Escherichia coli

The objective of this field trial was to determine if vaccination against Haemophilus parasuis serovar 5 (HPS 5) and pathogenic serotypes of Escherichia coli would improve nursery pig performance in an outdoor unit in different seasons. The unit was concurrently infected with HPS 5 and with different serotypes of E. coli. All piglets were born to HPS 5 vaccinated sows. The trial was carried out in four (two summer and two winter) groups. Group 1 (E. coli and HPS vaccinated, summer season) (n = 362): Piglets were vaccinated pre‐weaning with inactivated E. coli‐VT2e‐toxin and post‐weaning against HPS 5. Group 2 (non‐vaccinated, summer season) (n = 349): Piglets were not vaccinated. Group 3 (E. coli and HPS vaccinated, winter season) (n = 358): The animals were analogously treated as Group 1. Group 4 (non‐vaccinated, winter season) (n = 353): Piglets were not vaccinated. The following parameters were evaluated: A: average daily nursery weight gain (ADG), B: nursery mortality, C: feed efficiency (FE). No significant weight differences were detected within the vaccinated and non‐vaccinated summer or winter raised groups of weaners. Summer raised weaners were significantly (P<0.05) heavier from day 35 on than winter raised animals. ADG and FE of summer raised pigs were significantly better (weeks 1–3 P<0.05; fourth week post‐weaning P<0.01) during the nursery period than that of the winter raised groups. Winter raised vaccinated weaners showed during the last week of nursing significantly (P<0.05) better daily gain and feed efficiency compared with the non‐vaccinated winter raised animals. Non‐significant ADG and FE differences were detectable between the summer raised vaccinated or non‐vaccinated groups of pig. Winter raised non‐vaccinated animals suffered significantly (P<0.05) higher nursery mortality (10.63%) compared to the winter raised vaccinated animals. Implication: In cases of concurrent infections with HPS 5 and with different serotypes of E. coli, especially during winter season, vaccination against both diseases is suggested.     

Yeast-expressed classical swine fever virus glycoprotein E2 induces a protective immune response

Classical swine fever (CSF) is an economically important swine disease worldwide. The glycoprotein E2 of classical swine fever virus (CSFV) is a viral antigen that can induce a protective immune response against CSF. A recombinant E2 protein was constructed using the yeast Pichia pastoris expression system and evaluated for its vaccine efficacy. The yeast-expressed E2 (yE2) was shown to have N-linked glycosylation and to form homodimer molecules. Four 6-week-old specified-pathogen-free (SPF) piglets were intramuscularly immunized with yE2 twice at 3-week intervals. All yE2-vaccinated pigs could mount an anamnestic response after booster vaccination with neutralizing antibody titers ranging from 1:96 to 1:768. Neutralizing antibody titers at 10 weeks post booster vaccination ranged from 1:16 to 1:64. At this time, the pigs were subjected to challenge infection with a dose of 1 × 10⁵ TCID₅₀ (50% tissue culture infective dose) virulent CSFV strain. At 1 week post challenge infection, all of the yE2-immunized pigs were alive and without symptoms or signs of CSF. Neutralizing antibody titers at this time ranged from 1:4,800 to 1:12,800 and even to 1:51,200 one week later. In contrast, the control pigs continuously exhibited signs of CSF and had to be euthanized because of severe clinical symptoms at 6 days post challenge infection. All of the yE2-vaccinated pigs were E^rns antibody negative and had seroconverted against E^rns by post challenge day 11, suggesting that yE2 is a potential DIVA (differentiating infected from vaccinated animals) vaccine. The yeast-expressed E2 protein retains correct immunogenicity and is able to induce a protective immune response against CSFV 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.