Bull. Anim. Hlth. Prod. Aft.. (2007), 55, 5-11
FIELD EVALUATION OF THE POLYVALENT INACTIVATED BVD, IBR, PI-3 AND
BRSV VACCINE ADJUVANTED WITH NIGELLA SATIVA OIL IN PREGNANT DAMS
AND THEIR CALVES
M. H. Ebeid', A. A. A. El-Sawalhyz, EI-Sabbagh, M. M. A.Nabila3, Sh. Degheidy4 and
A. M. M. Allam4
'Faculty of Veterinary Medicine, Benha University Egypt
2 Faculty
of Veterinary Medicine, Mansoura University Egypt
'Veterinary Serum and Vaccine Research Institute Cairo Egypt
National Research Center Cairo Egypt
EVALUATION SUR LE TERRAIN DUN VACCIN INACTIVE POLYVALENTAVEC UN
ADJUVANT D'HUILE DE NIGELLE CONTRE LES VIRUS DE BVD, IBR, PI-3 ET BRS
CHEZ DES MERES GRAVIDES FT LEURS VEAUX
Resume
On a evalue choz dos mares gravides un vaccin inactive polyvalent avec un adjuvant dhuile do nigolle
contra le virus de la diarrhpe virale bovine (BVD), de la rhinotracheite inioctiouse bovine (IBR). le virus
paragrippal type 3 (PI-3) et to virus rospiratoire syncytial bovin (BRS). Les mCres etaient au dernier
stade de gravidito (troisieme trimestre). Trois semaines apres la vaccination. I'anticorps neutralisant a
commence a augmenter audessus du niveau de protection pouratteindre un niveau maximal au
velage. Les n&eaux de protection apres la deuxieme vaccination choz Los mares 1-1 ,4 mois apres le
velage etaient comme suit: 1.70: 1,80 :1,90 of 1,65 pour los virus de BVD, IBR, PI-3 of BRS
respectivement. Les niveaux de protection apres la deuxieme vaccination chez les mores 2-3 mois
apres le velago otaient : 1,95:2: 2.10 of 2 pour les virus do BVD, IBR. P1-301 BRS respectivement.
Apres to deuxieme vaccination des mores. les niveaux de protection des veaux issus de ces mares 11! a mois apres 10 velago 6taient comme suit : 1.25 ; 0.80: 0.55 of 0.60 pour les virus BVD. IBR, PI-3
of BRS respectivement. Apres la douxieme vaccination des mores. les niveaux de protection des veaux
issus do cos mares 2-3 mois apres le vclage etaient les suivants: 1,68:0.90:0.43 of 0.74 pour les virus
BVD. IBR, PI-3 et BRS respectivement. Le vacpn avec I' adjuvantd'huile do nigelle peut @tre utilise en
toute seoudte pour ('immunisation des mares gravides pour Is contrdte de ('infection des veaux
nouvellement mis bas.
Mots-cle : Vaccin, BVD, IBR, P13, BRS, huile de nigelle. bovins.
Summary
A polyvalent inactivated respiratory virus vaccine adjuvanted with Nigella saliva oil containing bovine
viral diarrhea (BVD). Infectious Bovine Rhinotracheitis (IBR), Parainfluenza typo 3 (PI-3) and bovine
respiratory syncytial virus (BRSV) was evaluated in pregnant dams. Dams were at late stage of
pregnancy (third trimester). The neutralizing antibody started to increase following vaccination at 3
weeks post vaccination above the protective level and increase to maximal level at parturition as
follows for dams delivered after 2nd vaccination during 1-1.5 months were 1.70, 1.80. 1.90. and 1.65 for
BVD. IBR. PI-3 and BRS viruses respectively. Dams delivered after 2nd vaccination during 2-3 months
were 1.95.2.00.2.10, and 2.00 for BVD. IBR. PI-3 and BRS viruses respectively. The titers of the
offspring born to vaccinated cows are as follows for calves delivered after 2nd vaccination during 1-1.5
months were 1.25.0.80.0.55, and 0.60 for BVD. IBA, PI-3 and BRS viruses respectively. Calves
delivered after 2nd vaccination during 2-3 months were 1.68, 0.90.0.43. and 0.74 for BVD, IBR, P1 .3
and BRS viruses respectively. The Nigella Saliva oil adjuvanted vaccine can be used safely in
immunization of pregnant dams to control the infection in newly borne calves.
Key-words :
Vaccine, BVD. IBR, PI-3. BRSV. Nigella Saliva oil and cattle
'corresponding Author a-mail: ahmedolSawalhya@yah00.COm
6
M. H. EBEID. A. A. A. EL-SAWALHY. M. M. A. EL-SABBAGH. NABILA. SH. DEGHEIDY and A. M. M. ALLAM
Introduction
Bovine respiratory diseases (BRD) have an important and serious impact on the beef cattle
industry, both for stocker and feedlot entities. Economic losses result from death, decreased
performance of diseased cattle, lowered weight gain, increased cost of gain, reduced carcass
value and treatment costs'.
BVD, IBR, PI-3 and BRS viruses have all been incriminated in the etiology of acute respiratory
diseases in cattle and calves feedlot' . They are causing serious losses each year due to the
controlling of BRD which is a major focus of veterinary health programs'. So. prevention and
control of BRD is much more successful and economically feasible than treatment. An ideal
processing protocol should include vaccination of the herd with safe, potent and effective vaccine
to develop an adequate immune response' 5.
Active immunity by natural infections or vaccinations may prolong persistence of maternally
derived antibodies. Vaccination and induction of protective immunity in calves at the earliest age is
ideal for prevention of future disease. Potentially boostering the cows annually with certain
vaccines might result in transfer of high levels of antibodies to the calves',. Nigella Saliva oil
adjuvanted vaccine is better and effective than other oil used in the experiment . (Marcol-52 and
Paraffin). This refers to its high immunogenic effect. It is natural, and produced locally in Egypt
when compared with the alhydrogel.
Therefore the present work aimed to study the duration of immunity for the Nigella Saliva oil
adjuvanted vaccine in field calves delivered from pregnant dams whichvaccinated with this
vaccine.
Materials and Methods
Materials
Animals
Cows
Ten apparently healthy pregnant cows were used for field evaluation of the prepared vaccine.
They were at third trimester of pregnancy. Cows were belonged to private farm located in
Domiatta city, Domiatta governorate, Egypt. Seven cows were kept for evaluation of the vaccine
and three pregnant cows were kept as non-vaccinated control animals.
Viruses
a. BVD virus, Iman strain (106.5 TCID50ml).
b. BHV-lvirus. Abou Hammad strain (107.5 TCID50/ml).
c. P13 virus, strain 45 (108 TCID50/ml).
d.
BRS virus, strain 375L (106.5 TCID50ml).
Samples
Serum
Serum samples were collected for evaluation of prepared vaccines. They were collected from
all calves and pregnant cows before and after vaccination. Calves' serum samples were collected
until antibodies titer became inprotective. In cows, offsprings' serum samples were collected after
ingestion of colostrums from calves (post-colostral samples). Then, calves were periodically
examined for 7 months post delivery for detection of antibodies for the vaccine.
M. H. EBEID. A. A. A. EL-SAWALHY. M. M. A. EL-SABBAGH. NABILA, SH. DEGHEIDY ant
A. M. A4. ALLAM
Methods
1. Serum neutralization test, serum samples were tested by SNTaccording to Rossi and Kiessell.
2- Field evaluation of the vaccine adjuvanted by Nigella Sativa oil in pregnant dams
Ten apparently healthy pregnant cows were used for field evaluation of the prepared
polyvalent inactivated vaccine adjuvanted with Nigella saliva oil. They belonged to private farm
located in Domiatta city, Domiatta governorate. They were at late stage of pregnancy (third
trimester) and divided as follows, seven pregnant cows were kept for evaluation of the vaccine
and three pregnant cows were kept as non-vaccinated control animals. Serum samples were
collected from cows before inoculationof the first dose of the vaccine, prior to administration of the
second dose of the vaccine and before parturition. In addition, serum samples were collected from
unvaccinated cows at the same intervals. Serum samples were collected from the newborn calves
at 1st week post delivery then every month till 7-month post delivery.
Statistical analysis
Data were analyzed using an analytical software program SPSS version 8.0 for Windows
(SPSS science. Chicago. II, USA) by using the non-parametric Mann-Whitney U test.
Results and Discussion
Pneumonic respiratory disease based on necropsy summary is the major cause of death in
stocker cattle and feedlot catField evaluation of the polyvalent inactivated BVD. IBR. P1 .3 and BRSV vaccine adjuvanted with
oil in pregn ant dams and their calves.
9 Nrge/ta Saliva
tl&. Vaccination programs for breeding herds, stocker operation and processing of cattle in feed
yards are integral parts of preventive health programs designed to lessen the effects of infectious
respiratory diseases in cattle.
Many authors used mineral oils as adjuvants in preparation of inactivated respiratory vaccines
and compared these adjuvants against the alhydrogel gel adjuvant'" .2. They found that the vaccinated calves against these respiratory pathogens with vaccines adjuvanted by mineral oils
showed a better neutralizing antibodies response to the oil adjuvant vaccines than to the
aluminum gel vaccines.
The vaccines with Nigella sativa oil adjuvant are of good quality and immunostimulant more
than vaccines with Marcol oil adjuvant'. The seeds of Nigella sativa contain both fixed and
essential oils, proteins, alkaloids and saponin. The extract —as a natural oil adjuvant - showed
antibacterial, antioxidant, analgesic, anti-inflammatory, and antifungal effect" t`. In addition, there
was a significant enhancement of phagocytosis and increase of the T helper cell population"".
A field experiment was performed to evaluate the polyvalent inactivated respiratory vaccine
adjuvanted with Nigella Sativa oil. The field condition was achieved in a private farm located in
Domiatta Governorate.
All cows in the vaccinated group and their offspring were normal during the experimental
period and did not exhibit any rise in body temperature. They did not show any signs of respiratory
disorders. This might be contributed to the safety of the prepared vaccine.
The neutralizing antibody started to increase following vaccination at 3 weeks post vaccination
above the protective level and
10
M. H. EBEID. A. A. A. EL-SAWALHY. M. M. A. ELSABBAGH. NABILA, SH. DEGHEIDY and A. M. M. ALLAM
increase to maximal level at parturition as
months when it comes from vaccinated
follows for dams delivered after 2nd vacci- dams with inactivated vaccine at the last
nation during 1-1.5 months (Table 1) were 1.70, 1.80, 1.90, and 1.65 for BVD, IBR. PIgestation period of pregnancy.
So we can conclude that the Nigella
3 and BRS viruses respectively. Dams de- Sativa oil adjuvanted vaccine can be used
livered after 2nd vaccination during 2-3 months (Table 1) were 1.95.2.00, 2.10, and 2.00 for BVD.
IBR, PI-3 and BRS viruses respectively.
The neutralizing antibody to offspring born from vaccinated cows are as follows for calves
delivered after 2nd vaccina tion during 1-1.5 months (Table 1 & Fig 1) were 1.25, 0.80, 0.55, and
0.60 for BVD, IBR, PI-3 and BRS viruses respectively. Calves delivered after 2nd vaccination
during 2-3 months (Table 1 & Fig 2) were 1.68, 0.90, 0.43, and 0.74 for BVD, IBR, PI-3 and BRS
viruses respectively.
These results are in disagreement with that obtained by Baz16 who reported that calves get the
same titer of antibodies or higher than their dams. The antibodies gradually decreased towards
the seventh month post parturition and reached to the minimal protective level of immunity.
These results agreed with that obtained by''s " who concluded that the maternal antibodies of
cattle calves that were developed because of vaccination of their dams with killed vaccine
containing BVD, IBR. PI-3 and BRS viruses might persist for 6 months.
Cattle calves that were delivered at 3 months of vaccination of their dams remained protected
till the 4", month as obtained by Brar2' and EI-Sabbagh~. They concluded that passive colostral
antibodies lost one-half of their remaining antibody titer eve ry 21 days and serologically
responded to vaccine. Thus, the present study indicated the maternal antibodies in calves remained within the protective level from 3-6safely in immunization of pregnant dams to control the
infection in newly borne calves.
References
1.
Fulton, R. W.: Ridpath. J. F.; Saliki, J. T.; Briggs, R. E.; Confer, A. W.: Burge. L. J.; Purdy,
C. W.: Loan. R. W.; Duff. (3. C. and
Payton, M. E. (2002). Bovine viral diarrhea virus (BVDV) 1b: predominant BVDV subtype in calves with
respiratory disease. Can J. Vet. Res.. Jul:66(3):181-190.
2.
Durham. P.J. K. and Hassard, L. E. (1990). Prevalence of antibodies to IBR, PI3. BRSV and BVD in cattle in
Saskatchewan and Alberta. Cand. Vet. J. 31: 815-820.
3.
Samira, S.T.; EI-Sabbagh. M.M.A. and Ghaly, H.M. (2001 b). Preparation of combined inactivated BVD, IBR. P13
and respiratory syncytial vines (BRSV). J. Egypt. Vet. Med. Ass., 61. (4) 251-263.
4, Slokka. GL. and EdvardsAlvin. L. (1990). Revaccination of stressed calves with a multiple polyvalent vaccine (IBR, BVD,
BRSV). Agr. Pract, 11 (5):18-20.
5.
Samira, S.T.; EI-Sabbagh, M.M.A.: Ghaly. H.M. and Abdel Samae, M. M. (2001 a). Production and evaluation of a
mullicomponant inactivated respiratory viral vaccine [BVDV IBRV. PI3V and bovine adeno virus type 3: Pneumo4] in cattle. Egypt. J. Agnc. Res. 79 (1): 335-351.
6.
10
Fulton, R. W.: Briggs, R. E.; Payton. M. E.: Confer, A. W.; Saliki, J.T.: Ridpath, J. F.; Burge. L. J. and Duff, G C.
(2004). Maternally derived humoral immunity to BVDV l a, BVDV1b, BVDV2. BHV-1. PI3V, BRSV, Mannhemia
haemolybca and Pasteurella multocida in beef calves, antibody decline by hall-life studios and effect on response to
vaccination. Vaccine. 26:22(5.6): 643649.
Ebeid. M. H.; El-Sawalhy, A. A. A.: ElSabbagh, M. M. A.: Nabila. Sh. Degheidy and Allarn, A. M. M. (2006).
Preparation of polyvalent inactivated BVD, IBR, PI-3 and
M. H. EBEID. A. A. A. EL-SAWALHY. M. M. A. ELSABBAGH. NABILA, SH. DEGHEIDY and A. M. M. ALLAM
increase to maximal level at parturition as
months when it comes from vaccinated
follows for dams delivered after 2nd vacci- dams with inactivated vaccine at the last
nation during 1-1.5 months (Table 1) were 1.70, 1.80, 1.90, and 1.65 for BVD, IBR. PIgestation period of pregnancy.
So we can conclude that the Nigella
3 and BRS viruses respectively. Dams de- Sativa oil adjuvanted vaccine can be used
livered after 2nd vaccination during 2-3 months (Table 1) were 1.95.2.00, 2.10, and 2.00 for BVD.
IBR, PI-3 and BRS viruses respectively.
The neutralizing antibody to offspring born from vaccinated cows are as follows for calves
delivered after 2nd vaccina tion during 1-1.5 months (Table 1 & Fig 1) were 1.25, 0.80, 0.55, and
0.60 for BVD, IBR, PI-3 and BRS viruses respectively. Calves delivered after 2nd vaccination
during 2-3 months (Table 1 & Fig 2) were 1.68, 0.90, 0.43, and 0.74 for BVD, IBR, PI-3 and BRS
viruses respectively.
These results are in disagreement with that obtained by Baz 16 who reported that calves get the
same titer of antibodies or higher than their dams. The antibodies gradually decreased towards
the seventh month post parturition and reached to the minimal protective level of immunity.
These results agreed with that obtained by''s " who concluded that the maternal antibodies of
cattle calves that were developed because of vaccination of their dams with killed vaccine
containing BVD, IBR. PI-3 and BRS viruses might persist for 6 months.
Cattle calves that were delivered at 3 months of vaccination of their dams remained protected
till the 4", month as obtained by Brar2' and EI-Sabbagh~. They concluded that passive colostral
antibodies lost one-half of their remaining antibody titer eve ry 21 days and serologically
responded to vaccine. Thus, the present study indicated the maternal antibodies in calves remained within the protective level from 3-6safely in immunization of pregnant dams to control the
infection in newly borne calves.
References
4.
Fulton, R. W.: Ridpath. J. F.; Saliki, J. T.; Briggs, R. E.; Confer, A. W.: Burge. L. J.; Purdy,
C. W.: Loan. R. W.; Duff. (3. C. and
Payton, M. E. (2002). Bovine viral diarrhea virus (BVDV) 1b: predominant BVDV subtype in calves with
respiratory disease. Can J. Vet. Res.. Jul:66(3):181-190.
5.
Durham. P.J. K. and Hassard, L. E. (1990). Prevalence of antibodies to IBR, PI3. BRSV and BVD in cattle in
Saskatchewan and Alberta. Cand. Vet. J. 31: 815-820.
6.
Samira, S.T.; EI-Sabbagh. M.M.A. and Ghaly, H.M. (2001 b). Preparation of combined inactivated BVD, IBR. P13
and respiratory syncytial vines (BRSV). J. Egypt. Vet. Med. Ass., 61. (4) 251-263.
4, Slokka. GL. and EdvardsAlvin. L. (1990). Revaccination of stressed calves with a multiple polyvalent vaccine (IBR, BVD,
BRSV). Agr. Pract, 11 (5):18-20.
7.
Samira, S.T.; EI-Sabbagh, M.M.A.: Ghaly. H.M. and Abdel Samae, M. M. (2001 a). Production and evaluation of a
mullicomponant inactivated respiratory viral vaccine [BVDV IBRV. PI3V and bovine adeno virus type 3: Pneumo4] in cattle. Egypt. J. Agnc. Res. 79 (1): 335-351.
8.
Fulton, R. W.: Briggs, R. E.; Payton. M. E.: Confer, A. W.; Saliki, J.T.: Ridpath, J. F.; Burge. L. J. and Duff, G C.
(2004). Maternally derived humoral immunity to BVDV l a, BVDV1b, BVDV2. BHV-1. PI3V, BRSV, Mannhemia
haemolybca and Pasteurella multocida in beef calves, antibody decline by hall-life studios and effect on response to
vaccination. Vaccine. 26:22(5.6): 643649.
Ebeid. M. H.; El-Sawalhy, A. A. A.: ElSabbagh, M. M. A.: Nabila. Sh. Degheidy and Allarn, A. M. M. (2006).
Preparation of polyvalent inactivated BVD, IBR, PI-3 and