The Veterinary Journal xxx (2013) xxx–xxx

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The Veterinary Journal

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A study of the effectiveness of a needle-free injection device comparedwith a needle and syringe used to vaccinate calves against bovine viraldiarrhea and infectious bovine rhinotracheitis viruses

1090-0233/$ - see front matter � 2013 Elsevier Ltd. All rights reserved.http://dx.doi.org/10.1016/j.tvjl.2013.06.019

⇑ Corresponding author. Tel.: +1 204 4747628.E-mail address: k_ominski@umanitoba.ca (K.H. Ominski).

Please cite this article in press as: Rey, M.R., et al. A study of the effectiveness of a needle-free injection device compared with a needle and syringevaccinate calves against bovine viral diarrhea and infectious bovine rhinotracheitis viruses. The Veterinary Journal (2013), http://dx.doi.org/1j.tvjl.2013.06.019

Michel R. Rey a, Michael Undi a, Juan C. Rodriguez-Lecompte b, Tomy Joseph c,d, Jason Morrison e,Alexander Yitbarek a, Karin Wittenberg a, Robert Tremblay f, Gary H. Crow a, Kim H. Ominski a,⇑a Department of Animal Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canadab Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, 500 University Avenue, Charlottetown, PEI C1A 4P3, Canadac Veterinary Diagnostic Services, MAFRI, Agricultural Services Complex, 545 University Crescent, Winnipeg, Manitoba R3T 5S6, Canadad Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba R3E 0J9, Canadae Department of Biosystems Engineering, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canadaf Boehringer Ingelheim (Canada) Ltd., Burlington, Ontario L7L 5H4, Canada

a r t i c l e i n f o a b s t r a c t

Article history:Accepted 26 June 2013Available online xxxx

Keywords:Bovine viral diarrhea virusInfectious bovine rhinotracheitis virusAntibody responseNeedle and syringeNeedle-free vaccination

The aim of this study was to compare the effectiveness of a needle-free injection device (NF) with a nee-dle and syringe (NS) when used to vaccinate calves against bovine viral diarrhea virus (BVDV) and infec-tious bovine rhinotracheitis virus (IBRV). The study was conducted in two independent phases. Ninety-six crossbred beef calves were vaccinated in the spring and 98 beef calves in the autumn. The calves werevaccinated using a NF or NS at 2 months of age (day 0) and again on day 119, with a modified-live virusvaccine containing IBRV, BVDV (types 1 and 2), parainfluenza-3 virus, and bovine respiratory syncytialvirus. In each herd 10 calves were left unvaccinated to determine whether exposure to either BVDV orIBRV occurred.

Visible vaccine residue at the surface of the skin/hair was apparent immediately following vaccinationwith NF in 30% of the spring-born calves following both the primary and booster vaccination. In theautumn, visible vaccine residues occurred in 19% and 8% of NF-vaccinated calves following the primaryand booster vaccination. Post-vaccination skin reactions recorded on days 21, 42, 119 and 140 occurredwith greater frequency in NF-vaccinated calves than NS-vaccinated ones.

Blood samples were collected on days 0, 21, 42, 119, and 140 and tested for antibodies to BVDV andIBRV. Vaccination technique had no significant effect on BVDV or IBRV antibody concentrations at anytime point. NF was as effective as NS vaccination in eliciting BVDV and IBRV antibody responses.

� 2013 Elsevier Ltd. All rights reserved.

Introduction

Vaccines have traditionally been administered using a needleand syringe (NS) but this can result in accidental injury of individ-uals handling needles (Weese and Jack, 2008) as well as the risk ofbroken needle fragments in meat (van Drunen Littel-van den Hurk,2006). Furthermore, blood-borne infectious diseases such as bo-vine leukosis (Hollis et al., 2005) and anaplasmosis (Reinboldet al., 2010) can be transmitted between animals if a single needleis used to inject multiple animals. These disadvantages have led tothe development of alternative vaccination techniques, includingthe use of needle-free injection devices (NFs).

NFs use mechanical compression that is triggered when thenozzle touches the skin, powering vaccine injections through asmall orifice with a high pressure stream that can penetrate theskin and deposit the vaccine into the desired tissue (Mouselet al., 2008). The technique eliminates broken needles, needle dis-posal and needle-stick injuries (Chase et al., 2008), reduces diseasetransfer (Reinbold et al., 2010) and vaccination time (Mousel et al.,2008), and provides greater antigen dispersion (Bennett et al.,1971). They have been used successfully to vaccinate humans(Hingston et al., 1963) and swine (Chase et al., 2008). Their usein cattle is currently limited but research regarding the use ofNFs in cattle has evaluated the immune response after vaccinationagainst Mannheimia haemolytica and Leptospira pomona (Holliset al., 2005), Brucella abortus (Pires et al., 2007), IBRV (Holliset al., 2005; van Drunen Littel-van den Hurk, 2006) and foot-and-mouth disease virus (Pandya et al., 2012).

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2 M.R. Rey et al. / The Veterinary Journal xxx (2013) xxx–xxx

The purpose of the present study was to compare the effective-ness of a NF and NS when used to vaccinate calves with a commer-cially-available, modified-live virus (MLV) combination vaccine(Express 5, Boehringer Ingelheim Canada) containing infectious bo-vine rhinotracheitis virus (IBRV), bovine viral diarrhea virus(BVDV) (types 1 and 2), parainfluenza-3 virus (PI-3), and bovinerespiratory syncytial virus (BRSV).

Materials and methods

Experimental animals and initial screening

Two independent trials were conducted in separate commercial cow–calf beefherds located in Manitoba, Canada. The first trial was conducted in the spring with96 crossbred beef calves (106.6 ± 16.8 kg), while the second trial was conducted inthe autumn with 98 calves (100.7 ± 16.2 kg). Prior to the start of each trial, bloodsamples were collected from calves via jugular venepuncture into sodium heparinvacutainer tubes (BD), and tested for BVDV antigen via RT-PCR at the VeterinaryDiagnostic Services Laboratory, Manitoba Agriculture, Food and Rural Initiatives.All calves were negative for BVDV.

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Vaccination protocol

In each trial, calves were randomly assigned to one of three groups: (1) vacci-nation using a NF, (2) vaccination with a NS, and (3) unvaccinated control group.In the spring group, 47, 39, and 10 calves were allocated to the NF, NS, and controlgroups, respectively. In the autumn, 48, 40, and 10 calves were allocated to thesame respective groups. Calves in the NF and NS groups were vaccinated at approx-imately 2 months of age (day 0) with the MLV combination vaccine, and then againon day 119, according to the manufacturer’s recommendation.

The NF used was a Pulse 250 NeedleFree Injection System which used com-pressed CO2 as the energy source. Cold weather made it necessary to use com-pressed N2 as the power source for the booster vaccination (day 119) in theautumn. The vaccine was administered on the left side of the neck using a skin-tenting technique, with NF set at 45–50 PSI and 85 PSI on day 0 and 119, respec-tively, in order to perform a subcutaneous (SC) injection (Pulse NeedleFree Sys-tems). Immediately following each NF vaccination, the presence of vaccineresidue on the skin/hair surface was recorded.

In the NS group, vaccinations were administered SC with a multi-dose pistol-grip syringe (Kane Veterinary Supplies), fitted with an 18 G, 2.54 cm (1 in.) detect-able needle (Partnar Animal Health). The same vaccine technique was used as forthe NF vaccination. To simulate standard industry practice, needles were changedonce every 10 calves, unless a needle became bent in which case it was immediatelyreplaced. The injected area of all vaccinated calves was inspected, palpated andscored on days 21, 42, 119 and 140 for the presence of skin reactions. Any apparentraised surface at the injection site was recorded as a vaccination reaction.

Animal feeding and management during both trials followed standard industrypractices. Animal handling and care procedures in this study were approved by theUniversity of Manitoba Animal Care Committee in compliance with the guidelinesof the Canadian Council on Animal Care (1993).

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Antibody response

Blood samples were collected via jugular venepuncture into serum vacutainertubes (BD) on days 0, 21, 42, 119 and 140. Samples were allowed to clot and thencentrifuged at 1100 g for 10 min. Aliquots of serum were stored at �20 �C untilanalysis for BVDV and IBRV antibody concentrations using commercial semi-quan-titative ELISA tests (BVDV Total Ab Test Kit and IBRV Individual Ab Test Kit, IDEXXLaboratories), following the manufacturer’s instructions. Antibody concentrationswere expressed as a sample to positive ratio (S/P Ratio) calculated as: S/PRatio = (Sample optical density (OD) � Negative control mean OD)/(Positive controlmean OD � Negative control mean OD). The S/P Ratio was transformed to a log10(S/P Ratio + 1) scale prior to analysis.

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Fig. 1. BVDV antibody levels [log10(S/P Ratio + 1)] in calves vaccinated using NF andNS devices in the spring (a) and autumn (b). There were 10 calves in the controlgroup. Arrows (;) indicate the day calves were vaccinated.

Statistical analysis

The spring and autumn trials were analyzed separately. A linear mixed repeatedmeasures model (SAS 9.2; SAS Institute) was used to analyze antibody concentra-tions with fixed effects of group, days post-vaccination and their interaction andanimal within group as a random effect. Two calves, one from the NF group inthe spring and one from the NS group in the autumn, were removed from the studyfor reasons unrelated to the study. Confidence limits (95%) of the estimated fre-quencies of skin reactions for each group at each time point were calculated to eval-uate the effect of vaccination device on such reactions. An error in data collectionmeant that skin reaction data was not available on days 21 and 42 of the springtrial.

Please cite this article in press as: Rey, M.R., et al. A study of the effectiveness ofvaccinate calves against bovine viral diarrhea and infectious bovine rhinotraj.tvjl.2013.06.019

Results

Vaccine residues

Visible vaccine residue was observed in approximately 30% ofNF-vaccinated calves following both primary and booster vaccina-tions in the spring. In the autumn, visible vaccine residue occurredin 19% and 8% of NF-vaccinated calves following primary and boos-ter vaccinations, respectively.

BVDV antibody response

BVDV antibody concentrations were not affected by vaccinationtechnique, nor was there an interaction between time and device(P > 0.05). Changes in BVDV antibody concentrations in NF- andNS-vaccinated calves were similar in the spring (Fig. 1a) and au-tumn (Fig. 1b).

IBRV antibody response

IBRV antibody concentrations were not affected by vaccinationtechnique (P > 0.05). Concentrations of IBRV antibodies were high-er (P < 0.05) in NF-vaccinated calves than NS-vaccinated calves ondays 42 and 119 in the spring (Fig. 2a) and day 42 in the autumn(Fig. 2b). In both trials, IBRV antibody concentrations decreasedprogressively following the initial vaccination, a trend that closelyfollowed that of control calves.

a needle-free injection device compared with a needle and syringe used tocheitis viruses. The Veterinary Journal (2013), http://dx.doi.org/10.1016/

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Fig. 2. IBRV antibody levels [log10(S/P Ratio + 1)] in calves vaccinated using NF andNS devices in the spring (a) and autumn (b). There were 10 calves in the controlgroup. NF and NS means within day with a different letter differ (P < 0.05). Arrows(;) indicate the day calves were vaccinated.

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Fig. 3. Percentage of skin reactions (mean ± 95% confidence limits) in calvesvaccinated against BVDV and IBRV using NF and NS devices in the spring (a) andautumn (b). Means within day with a different letter differ (P < 0.05).

M.R. Rey et al. / The Veterinary Journal xxx (2013) xxx–xxx 3

Skin reactions

In the spring (Fig. 3a) and autumn (Fig. 3b), post-vaccinationskin reactions occurred with greater (P < 0.05) frequency in NF-vaccinated calves relative to NS-vaccinated calves.

Discussion

BVDV antibody concentrations in control calves declined overtime, indicating high levels of maternal antibodies at the start ofthe study and the absence of an immune response to wild BVDVduring the study. There was no detectable increase in BVDV anti-body concentrations at 21 days post-primary vaccination, probablydue to the high level of maternal antibodies on day 0. However,over the remaining period prior to the booster vaccination, BVDVantibody concentrations in NF- and NS-vaccinated calves werehigher than the controls, indicating that despite the high maternalantibodies on day 0, vaccination did elicit an active BVDV antibodyresponse, consistent with previous reports (Endsley et al., 2003;Kirkpatrick et al., 2008). Following the booster vaccination onday 119, both NF- and NS-vaccinated calves showed a significantantibody response. Over the whole study period, the antibody re-sponse was the same in vaccinated cattle irrespective of which de-vice was used.

IBRV antibody concentrations in both NF- and NS-vaccinatedcalves decreased following the primary vaccination but increased

Please cite this article in press as: Rey, M.R., et al. A study of the effectiveness ofvaccinate calves against bovine viral diarrhea and infectious bovine rhinotraj.tvjl.2013.06.019

following the booster vaccination. This response is similar to thatobserved in studies that have evaluated IBRV vaccination of calvesusing NS (Brar et al., 1978; Menanteau-Horta et al., 1985). The lackof an initial response is probably due to maternal antibody inter-ference with antibody production (Menanteau-Horta et al., 1985).However, this lack of response does not necessarily imply thatthe early IBRV vaccination was not important since it is likely thatthe increase following the second vaccination was an anamnesticresponse (Menanteau-Horta et al., 1985).

The IBRV antibody concentrations in the NF-vaccinated groupwere higher at some time points than those in the NS-vaccinatedgroup. Previous research in cattle has shown that vaccination withNFs resulted in comparable and sometimes enhanced immune re-sponse when compared to NS vaccination (Hollis et al., 2005; vanDrunen Littel-van den Hurk, 2006; Pires et al., 2007; Pandyaet al., 2012). Enhanced immune response following NF vaccinationmay be due to wider vaccine dispersion into tissues and greaterpenetration through the skin (Bennett et al., 1971). Furthermore,Langerhans cells present in the epidermis may enhance the im-mune response as they have high migratory mobility and are effec-tive antigen presentation cells (Bodey et al., 1997). This results inincreased inflammation, which causes recruitment of immune-competent inflammatory cells and allows for increased contactbetween the vaccine antigen and immune cells (Giudice andCampbell, 2006).

Post-vaccination skin reactions can be caused by several factors,including pressure trauma associated with vaccination or contam-ination by transmitting microorganisms from the skin/hair surface

a needle-free injection device compared with a needle and syringe used tocheitis viruses. The Veterinary Journal (2013), http://dx.doi.org/10.1016/

4 M.R. Rey et al. / The Veterinary Journal xxx (2013) xxx–xxx

into the tissue surrounding the injection site (Troxel et al., 1997).In the current study, NF vaccination caused a greater percentageof skin reactions than NS vaccination. The presence of skin reac-tions following NF vaccination may hinder the use of NFs if produc-ers believe that NF vaccination has adverse carcass quality effects.Future research should evaluate the impact of NF vaccination onskin reactions and relationship to carcass quality.

NF injection devices have been reported to leave residual vac-cine on the surface of the skin/hair, which may be unavoidableand may hinder the acceptance of the technology (Jones et al.,2005). Chase et al. (2008) proposed that the residual vaccine vol-ume observed at injection sites following NF vaccination is usuallyquite small (0.0004 mL). The presence of vaccine residue in NF-vac-cinated calves in the present study did not compromise antibodyresponse in NF-vaccinated calves.

Conclusions

The immune response after NF vaccination was comparable tothat after NS vaccination in both spring- and autumn-born calves.The higher percentage of post-vaccination skin reactions and thepresence of more vaccine residue in NF-vaccinated calves did notinfluence the effectiveness of the NF. Needle-free injection devices,such as the one evaluated in this study, can be used to vaccinatecalves against BVDV and IBR.

Conflict of interest statement

None of the authors has any financial or personal relationshipsthat could inappropriately influence or bias the content of thepaper.

Acknowledgements

The authors wish to thank Growing Forward, Manitoba Agricul-ture, Food and Rural Initiatives (MAFRI) for financial support. Spe-cial thanks are also due to Pulse NeedleFree Systems for providingthe needle-free injection device and to Boehringer Ingelheim (Can-ada) Ltd. for donation of the Express 5 vaccine. The authors are in-debted to Betty Green and family at G7 Ranch and the staff at EURRanch, who gratefully allowed the use of their cattle for the trialand for help on sampling days. The technical assistance providedby Terri Garner, Deanne Fulawka, Dana Gardiner, Carson Callum,Nicole Ireland, Mateo Remonda, Romina Livolsi, Ashley Rawluk,Sean Thompson, Harold Echeverry and Janice Haines is gratefullyacknowledged.

Please cite this article in press as: Rey, M.R., et al. A study of the effectiveness ofvaccinate calves against bovine viral diarrhea and infectious bovine rhinotraj.tvjl.2013.06.019

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a needle-free injection device compared with a needle and syringe used tocheitis viruses. The Veterinary Journal (2013), http://dx.doi.org/10.1016/