Acta agriculturae Slovenica, suplement 2 (september 2008), 167–171. 
 
16th Int. Symp. “Animal Science Days”, Strunjan, Slovenia, Sept. 17–19, 2008. 
  
Agris category codes: L50, L73 COBISS Code 1.08 
ANTIBODY RESPONSE FOLLOWING SOW VACCINATION USING CELL AND 
RECOMBINANT VACCINES, SINGLE AND MULTIPLE APPLICATION 
Stane KOŠOROK a) and Miran KASTELIC b) 
a) Farme Ihan d.d., Breznikova 89, SI-1230 Domžale, Slovenia 
b) Univ. of Ljubljana, Biotechnical Fac., Dept. of Animal Science, Groblje 3, SI-1230 Domžale, Slovenia 
ABSTRACT 
The vaccination of pregnant sows with E. coli antigens to increase specific antibodies in 
colostrum and milk and subsequently the protection of suckling piglets from neonatal E. coli 
diarrhea is a common and efficient method, already used for many years. The vaccination to 
protect individual animals against erysipelas has been used even for a longer time. There are 
many vaccines and vaccination techniques available on the market, among them herd specific 
“stable” vaccines for E. coli, inactivated cell vaccines and recombinant DNA vaccines. Unlike 
vaccines for pet animals, which are mostly polyvalent vaccines, there are very few polyvalent 
pig vaccines available on the EU market. Recently, Intervet company has developed Diluvac 
forte (DF) adjuvant which enables mixing of different monovalent vaccines. Porcilis DF vaccine 
line makes it possible to combine different antigens in the syringe and only one injection of 
desirable combination of antigens. The difference between two different vaccines (Colisorb, 
Porcilis DF) was tested to find out the antibody response following vaccination with monovalent 
recombinant vaccine against bivalent cell vaccine, and the antibody response followed separate 
and mixed injection of two antigens in one syringe. Statistical evaluation of serum antibody 
response obtained by ELISA test confirmed the expectations. DNA recombinant vaccines gave 
significantly higher antibody titers compared to cell vaccine. Porcilis vaccines mixed together 
prior to application gave surprisingly better response compared to separated application. The 
result leads to reduction of injections to pregnant sows and consequently to better immune 
response. 
Key words: pigs / sows / immunology / antibody response / vaccination / E .coli / erysipelas  
IMUNSKI ODZIV SVINJ PO VAKCINACIJI S CELIČNO IN REKOMBINANTNO 
VAKCINO Z LOČENO IN MEŠANO APLIKACIJO 
IZVLEČEK 
Vakcinacijo brejih svinj z antigeni E. coli uporabljamo že desetletja, da bi povečali specifična 
protitelesa v kolostrumu in mleku in tako pasivno zaščitili sesne pujske pred drisko, ki jo 
povzroča E .coli. Vakcinacija proti rdečici se uporablja še dlje. Na trgu je na voljo veliko 
komercialnih vakcin od t.i. hlevskih vakcin, inaktiviranih celičnih vakcin in v zadnjem času 
rekombinantnih DNK vakcin. Za razliko od vakcin, ki se uporabljajo pri malih živalih in ki so 
polivalentne, so na trgu EU predvsem monovalentne vakcine za prašiče. Nedavno je firma 
Intervet razvila DF (diluvac forte) adjuvant, ki omogoča mešanje različnih monovalentnih vakcin 
in s tem enkratno aplikacijo več poljubnih antigenov. Testirali smo razliko med dvema 
različnima vakcinama (Colisorb, Porcilis DF) z enakimi antigeni, da bi ugotovili, kakšen je 
imunski odziv po vakcinaciji z inaktivirano celično in rekombinantno vakcino in kakšna je 
razlika v količini protiteles po aplikaciji z ločenimi vakcinami v primerjavi z aplikacijo zmešanih 
antigenov. Statistična analiza nivoja protiteles po ELISA testiranju je potrdila naša pričakovanja. 
DNK rekombinantna vakcina daje značilno višje titre v primerjavi s celično vakcino. Porcilis DF 
vakcine, zmešane skupaj pred aplikacijo, presenetljivo spodbudijo boljši imunski odgovor kot 
http://aas.bf.uni-lj.si 
Acta agriculturae Slovenica, suplement 2 (september 2008). 
 
168 
ločena aplikacija. Rezultati testiranja kažejo, da je mogoče zmanjšati število aplikacij vakcin 
brejim svinjam in hkrati doseči boljši imunski odziv. 
Ključne besede: prašiči / svinje / imunologija / protitelesa / odziv / vakcinacija / E. coli / rdečica 
INTRODUCTION 
Neonatal Escherichia coli (E. coli) diarrhea is an important animal health concern for pig 
producers. It is responsible for significant economic losses. E. coli diarrhea is observed most 
commonly in piglets aged 1–4 days. Causative strains of E. coli are adhered by fimbrial adhesins 
(K88, K99, 987P and F41) (Hall, 1989; Nakazawa et al., 1987) to the small intestinal mucosa 
and produce one or more enterotoxins (ST, LT) (Bertschinger et al. 1992; Hall, 1989). Immunity 
to E. coli diarrhea is humoral and is initially provided via maternal colostrum. Specific 
antibodies inhibit bacterial adherence to the gut wall and neutralize the activity of the 
enterotoxins produced by the attached E. coli. Maternal vaccination has been one of the most 
effective ways of controlling neonatal E. coli diarrhea (Genovese et al., 2001; Osek et al., 1995). 
The application of recombinant DNA technology has resulted in more efficient vaccines (Riising 
et al., 2005; Larsen et al., 2004b; Moon and Bunn, 1993; Nagy, 1986; Clarke et al., 1985). 
Commercially available vaccines are given parenterally at about 6 and 3 weeks prior to 
parturition. These may be killed whole cell bacterins or purified fimbrial vaccines. 
The purpose of the trial was to determine whether new recombinant monovalent vaccines 
(Porcilis) give higher antibody response compared to the old cell vaccine (Colisorb). Secondly, 
the aim was to find out if different monovalent Porcilis vaccines containing DF (diluvac forte) 
adjuvants can be mixed together in the syringe prior to vaccination and obtain impaired results 
compared to separated vaccination (Larsen et al., 2004a). 
MATERIAL AND METHODS 
The purpose of the trial was to determine possible difference in blood serum antibody levels 
following vaccination with three different vaccines and two different application techniques. 
Tested vaccines were: Colisorb (Hoechst, Intervet, bivalent E. coli and erysipelas vaccine), 
Porcilis Coli DF (Intervet, monovalent E. coli vaccine) and Porcilis Ery DF (Intervet, 
monovalent erysipelas vaccine). On a commercial farm 120 pregnant sows and gilts (min. 5 gilts 
per group) inseminated in the same week were divided into three separated groups (A, B, C) 
containing 40 animals each. They were vaccinated at week 7 and week 3 prior to expected 
farrowing. On average 6 days before expected farrowing (approx. 15 days after the second – 
booster vaccination) the blood samples were collected and sent to the lab for antibody level 
determination. Group A: animals vaccinated with Colisorb 2 ml intramuscular (i/m). Group B: 
vaccinated with Porcilis coli and Porcilis ery separately 2 ml each vaccine – i/m. Group C: 
vaccinated with Porcilis coli and Porcilis ery mixed together in one bottle/syringe (DF adjuvant) 
4 ml i/m. Antibody response against the following antigens was tested with the following 
standard ELISA tests: ECO - K88ab, ECO - K88ac, ECO - K99, ECO - 987P, ECO - LT (labile 
toxin) and Erysipelothrix rhusiopathiae ELISA - ERY IRPC value (Cypress diagnostics). 
Laboratory analyses were done at Intervet R&D Service Lab, 5830 AA Boxmeer, The 
Netherlands. 
The data evaluation was done in two steps. In the first step the means and standard deviations 
for every experimental group (A, B and C) were estimated. In the second step the data were 
evaluated with the following statistical model: 
( ) ijkijkjiijk ePPbPGY +−+++= µ  
Košorok, S. and Kastelic, M. Antibody response following sow vaccination … and multiple aplication. 
 
169
where Yijk is an observed trait (adsorbance for every studied concentration of antibodies in blood 
serum), µ is an average value of the model, Gi is a fixed effect of the group (group A, B an C), Pj 
is a fixed effect of the parity, b is a linear regression coefficient of the time period between the 
date of vaccination and the date of farrowing and eijkl is the residual for the observation ijk. 
Means and standard deviations were estimated with SAS/BASIC procedure MEANS. The 
statistics for linear statistic model were estimated with SAS/STAT procedure GLM. 
RESULTS AND DISCUSSION 
The number of treated animals per group, the average value and standard deviations are 
presented in the Table 1. The number of observations laid between 34 and 38. The sizes of 
groups were relatively good weighted. The values for adsorbances, connected with immunity 
against different strains of E. coli, were between 8.89 and 13.64. The differences were larger 
between different strains than between different groups inside the same stain. The values for 
group A were generally slightly lower than the values for the other two groups.  
 
Table 1. Number ob observations per group (n), means and standard deviations (SD) for groups 
A, B and C for traits: period (time interval between the day of blood collection and 
farrowing day, in days), litter size, and adsorbances for antibodies of E. coli strains 
K88ab, K88ac, K99, 987p, for labile toxin (LT) and adsorbances for antibodies for 
erysipelas (Ery) 
 
group A group B group C Trait n mean SD n mean SD n mean SD 
period 36 9.44 1.75 38 8.89 2.61 37 9.27 1.22 
litter size 36 13.64 2.53 38 13.45 2.60 37 14.11 2.45 
K88ab 36 12.04 1.21 38 12.49 1.25 36 13.17 1.15 
K88ac 36 12.56 0.97 38 13.04 1.37 36 13.64 1.03 
K99 36 9.51 0.98 38 11.05 0.96 36 11.21 1.04 
987p 36 10.66 0.92 38 13.14 1.00 36 13.11 1.06 
LT 34 8.02 1.05 38 10.11 1.13 36 10.29 1.32 
Ery 36 116.73 37.43 37 157.13 27.14 36 158.49 22.80 
 
Standard deviations (SD) presented about 10% of average value. The group with older 
bivalent vaccine showed on average a little lower variability compared to both other groups. The 
cause of larger variability after group with the recombinant vaccine cannot be explained with the 
results from this experiment.  
The immunity against Erysipelothrix rhusiopathiae was also much larger in groups B and C 
than in group A. In that case, the variability following vaccination with the old vaccine was also 
much larger. 
In the Table 3 the results of analysis of variance according to the model described in chapter 
Material and methods are presented. The R2’s were middle to large that is between 0.33 and 
0.65.  
The group means the type of vaccine combined with the type of application. It statistically 
significantly influenced the quantity of antibodies measured as absorbance for every strain. The 
parity influenced the immunity against both strains K88 – ab and ac as well as the immunity 
against Erysipelothrix rhusiopathiae. The immunity against two other strains (K99 and 987p) did 
not depend on parity – the immunocompetence did not change during the productive life of sows 
and high immunity was expected also at the first parity. This was not the case for both K88 
Acta agriculturae Slovenica, suplement 2 (september 2008). 
 
170 
strains where the immunity increased from first to second parity. Some higher incidence of 
diarrhea in first little piglets can be expected because of lower immunity in gilts. The time 
interval between the time of vaccination and farrowing influenced only the immunity against 
strain 987p. In this particular case, late vaccination resulted in lower piglet protection. 
 
Table 2. Analysis of variance for observed adsorbaces for antibodys of E. coli strains K88ab, 
K88ac, K99, 987p, for labile toxin (LT) and adsorbances for antibodies for erysipelas 
(Ery) 
 
 model Effects 
 df    Ti Pj regresion 
Trait mod. error F P R2 df P df P df P b 
K88ab 10 99 4.96 <0.0001 0.3336 2 0.0004 7 0.0004 1 0.5212 – 0.0360
K88ac 10 99 5.96 <0.0001 0.3756 2 <0.0001 7 <0.0001 1 0.9226 0.0050
K99 10 99 7.10 <0.0001 0.4178 2 <0.0001 7 0.8157 1 0.0750 0.0918
987p 10 99 18.56 <0.0001 0.6521 2 <0.0001 7 0.1166 1 0.0007 0.1691
LT 10 97 12.62 <0.0001 0.5654 2 <0.0001 7 0.0018 1 0.0449 0.1109
Ery 10 98 7.19 <0.0001 0.4232 2 <0.0001 7 0.0106 1 0.3580 1.3310
 
The LSMEANS values for the group corresponded to the statistical model. The old bivalent 
vaccine (group A) generally caused lower immunoresponse compared to the new recombinant 
vaccines. Only in case of K88ab strain the group A was as good as group B, but both mentioned 
groups were worse than group C with mixed monovalent recombinant vaccines. 
 
Table 3. LSMEANS for groups A, B and C according to the statistical model* 
 
Trait group A group B group C 
K88ab 12.03a 12.48a 13.27 
K88ac 12.36 12.91 13.78 
K99 9.37 10.95a 11.22a 
987p 10.34 12.99a 13.09a 
LT 7.49 9.68 10.37 
Ery 118.59 161.66a 158.16a 
* the values with the same letter are not statistically significantly different (P > 0.05) 
 
The differences between the old bivalent vaccine and the recombinant new type of vaccines 
were expected. The group C – mixed monovalent recombinant vaccines surprisingly caused 
better immune response compared to the vaccination with the same but unmixed vaccines 
applied separately (group B). The effect is difficult to explain, but is known from the previous 
studies. 
CONCLUSIONS 
The newer recombinant vaccines against different E. coli antigens and against Erysipelothrix 
rhusiopathiae were compared with the old bivalent cell vaccine (group A). The recombinant 
vaccines were applied separately (group B) or they were mixed (group C). The recombinant 
vaccines caused stronger antibody response compared to the old bivalent vaccine. Group C was 
Košorok, S. and Kastelic, M. Antibody response following sow vaccination … and multiple aplication. 
 
171
in absolute values better than groups A and B. In cases of antibodies against K88ab and LT toxin 
the application of mixed vaccines resulted in higher antibody response than the application of 
two monovalent vaccines separately. In the other six cases the application of mixed or separated 
vaccines was equivalent. The effect of parity influenced the level of antibodies against strains 
K88ab, K88ac and Erysipelothrix rhusiopathiae as well as the concentration of LT. Higher 
incidence of diarrhea in first litter piglets was expected. The time period between the vaccination 
and the day of blood samples collection influenced only the level of antibodies against 987p 
strain and LT toxin. Vaccination with mixed recombinant vaccines is recommended due to 
powerful antibody response, less injections required and lower labor input. 
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