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Research Note
Field evaluation of the accuracy of vaccine deposition by two different
commercially available in ovo injection systems
ABSTRACT The location of injection and vaccine de- in which the vaccine was injected into the amniotic
position in ovo is known to be critical to the efficacy of sac or into s.c. or i.m. regions of the embryo. Incorrect
Mareks disease (MD) vaccine protection against MD vaccine delivery was defined as delivery into the air
viral challenge. Vaccine deposition into the amniotic cell; allantoic sac; any combinations including air cell
sac or a s.c. or i.m. site of the embryo is required for or allantois; the abdominal, cranial, orbital, or thoracic
MD vaccine efficacy. Vaccine deposition into the air cavities of the embryo; or no vaccine delivery at all.
cell or allantoic fluid results in chicks that are not ad- In hatchery 1 (Chick Master, Newton, MS) 1,171 nor-
equately protected against subsequent MD viral chal- mal eggs were processed through the Inovoject system
lenge. A study was conducted in 2 commercial broiler and 1,138 eggs were processed by the Intelliject system.
hatcheries to evaluate the ability of 2 in ovo injection The Inovoject system correctly vaccinated 94.62% of
systems, the Embrex Inovoject system (Pfizer Poultry the normal eggs as compared with 61.16% delivery ac-
Health, Research Triangle Park, NC) and the Intelliject curacy of normal eggs with the Intelliject system. In
system (Avitech, Salisbury, MD; distributed by Merial hatchery 2 (Jamesway Super J, Magee, MS) 926 normal
Ltd., Gainesville, GA) to deliver a vaccine approved eggs were processed by the Inovoject system and 910
for use in ovo accurately and properly. A standard MD normal eggs were processed by the Intelliject system.
vaccine diluent mixed with a protein-staining dye was The Inovoject system correctly vaccinated 91.04% of
delivered through each machine to simulate in ovo vac- the normal eggs, whereas the Intelliject system correct-
cination. The location of the dye within the egg deter- ly vaccinated 71.98% of the normal eggs. The results
mined whether the vaccine was delivered correctly. Each of this study clearly demonstrate that the Inovoject
egg was also evaluated for normal embryo development system accurately delivered in ovo vaccine at a signifi-
(normal eggs). Correct vaccine delivery included eggs cantly higher rate than the Intelliject system.
Key words: site of injection, in ovo, vaccine deposition, embryo
2011 Poultry Science 90:223226
doi:10.3382/ps.2010-00759
223
Injection system Correct delivery Incorrect delivery Correct delivery Incorrect delivery
all incorrect embryo sites (intracranial, intraorbital, tray to tray associated with eggs from the oldest breed-
intraabdominal). No attempts were made to quantify er flock. Only live normal embryos at injection time
or differentiate volume of vaccine delivered to multiple were included in the evaluation. The Inovoject system
sites in ovo; therefore, suboptimal doses or partial de- correctly injected a higher percentage of normal eggs
livery to correct sites in ovo were considered incorrect (P 0.0001) than the Intelliject system (Table 1). The
applications. Correct injections included only complete same model was then applied to each specific category
delivery of individual vaccine dispensed into the correct (vs. all others). The Intelliject system was significantly
site in ovo. The egg was considered the experimental higher (P 0.0001; Table 2) in all categories of incor-
unit in a generalized randomized complete block design rect vaccine deposition (AC, AL, AC or AL combina-
with flock as the blocking factor. Correct vs. incorrect tions, and EM incorrect), whereas the Inovoject system
injections were analyzed by exact conditional logistic performed significantly better in all categories of cor-
regression, with flock as a stratifying variable and treat- rect vaccine deposition (AM and EM proper) except for
ment as a factor (SAS/STAT Version 9.1.3 Service Pack the combination AM and EM injections, for which the
3, SAS Institute Inc., Cary, NC). In terms of adjust- Intelliject system delivered significantly more than the
ment for multiple comparisons, the overall treatment Inovoject system (P 0.0001).
effect for site of injection was evaluated using a Fishers In the Jamesway hatchery, 1,008 eggs were injected
protected least significant difference type approach for by each injection system. Of those, 926 normal eggs
comparisons for each site without adjustment. Statisti- were injected by the Inovoject system and 910 normal
cal significance was determined with P 0.05. eggs were injected by the Intelliject system. As in tri-
al 1, the Inovoject system (Table 3) had significantly
RESULTS AND DISCUSSION more correct vaccinations (AM) in ovo (P 0.0001)
than the Intelliject system, except for injections of EM
Two separate analyses were completed, one for each proper, for which there was no significant difference (P
type of incubation system involved, because myriad ran- > 0.5984) and injections of AM-EM proper, for which
dom effects were related to each hatchery and respec- the Intelliject system had significantly (P 0.0114)
tive incubation system. In the Chick Master hatchery, more proper injections than the Inovoject system. The
1,296 eggs were injected by each injection system. Of Intelliject system (Table 3) delivered significantly (P
those, 1,171 normal eggs were injected by the Inovoject 0.0001) more incorrect vaccine placements (AC, AC or
system and 1,138 normal eggs were injected by the In- AL combinations, and EM incorrect) when compared
telliject system. Although randomized at injection, the with the Inovoject system, except for AL (P > 0.4308)
difference in the number of normal eggs between the and the combination AM-EM incorrect injections (P >
2 groups was primarily due to fertility variation from 0.9999), for which there was no significant difference.
Table 2. Number and percentage of eggs at each specific site of vaccine deposition1 at the Chick Master hatchery
AC AL AL AM-EM EM Total
Item AC combination AL AM combination AM incorrect AM-EM incorrect EM eggs
Injection system
Embrex Inovoject
system
Eggs (no.) 0 3 1 2 0 673 1 2 56 433 1,171
Delivery (%) 0.00b 0.26b 0.09b 0.17b 0.00b 57.47a 0.09b 0.17b 4.78b 36.98a
Intelliject system
Eggs (no.) 41 26 18 179 78 480 20 48 80 168 1,138
Delivery (%) 3.60a 2.28a 1.58a 15.73a 6.85a 42.18b 1.76a 4.22a 7.03a 14.76b
Total eggs 41 29 19 181 78 1,153 21 50 136 601 2,309
a,bNumbers within a column lacking a common superscript differ (P 0.05).
1AC = air cell; AL = allantoic sac; AM = amniotic fluid; EM = embryo body.
Injection system
Embrex Inovoject
system
Eggs (no.) 2 2 27 48 3 744 0 3 1 96 926
Delivery (%) 0.22b 0.22b 2.92a 5.18b 0.32b 80.35a 0.00a 0.32b 0.11b 10.37a
Intelliject system
Eggs (no.) 17 30 33 134 30 541 1 13 10 101 910
Delivery (%) 1.87a 3.30a 3.63a 14.73a 3.30a 59.45b 0.11a 1.43a 1.10a 11.10a
Total eggs 19 32 60 182 33 1,285 1 16 11 197 1,836
a,bNumbers within a column lacking a common superscript differ (P 0.05).
1AC = air cell; AL = allantoic sac; AM = amniotic fluid; EM = embryo body.
With respect to accuracy of vaccine deposition by the lenge, whereas delivery to the AC shows no embryo
Inovoject system in the Chick Master hatchery vs. the access to the vaccine, thus no vaccinal response. As
JW hatchery, differences could be seen that were caused the embryo matures and begins the hatching process
by embryo age and egg setter tray design. The egg tray (internal and external pipping), more embryos in the
configuration and stability of the egg in the tray varied population are injected either s.c. or i.m. into the right
between the 2 incubation systems, with the Jamesway breast area. Embryo body delivery during in ovo vac-
incubation tray allowing eggs to deviate more than the cination is normal; however, penetration too deep into
Chick Master incubation tray along the upright axis. the egg and embryo during late-stage development can
The design of the Chick Master incubation tray kept cause excessive trauma to the embryo. Therefore, use
eggs in a very tight, upright position and did not al- of an in ovo injection system that does not deliver vac-
low egg movement laterally while the eggs were being cine to the correct embryonic site would compromise
turned in the incubator. In addition, embryos in the embryo safety and vaccine performance. The objective
Chick Master hatchery were 18 h older at transfer com- of commercial in ovo vaccination is to vaccinate every
pared with those in the Jamesway hatchery. The opti- viable embryo safely and uniformly. To accomplish this
mal time to inject eggs, developmentally, is the physi- objective, a commercial in ovo vaccination device must
ological stage between when the stalk of the yolk sac is administer vaccine to sites that result in the greatest
beginning its ascent into the abdomen and the head is safety and efficacy for the vaccine delivered. In this
tucked under the wing up until external pipping is initi- study, the Inovoject system was significantly better at
ated. The optimal target site for uniform automated in meeting that objective.
ovo vaccination is the AM or right breast area of the
developing embryo. Injections into the cranium, ocu- REFERENCES
lar space, and abdomen of the embryo are considered
off-target injections and are a result of misalignment Gildersleeve, R. P., C. M. Hoyle, A. M. Miles, D. L. Murray, C. A.
Ricks, M. N. Secrest, C. J. Williams, and C. L. Womack. 1993.
of injectors with the eggshell surface. These delivery Developmental performance of an egg injection machine for ad-
sites in ovo are not desirable for MD vaccine delivery ministration of Mareks disease vaccine. J. Appl. Poult. Res.
to the late-stage developing embryo and are incorrect 2:337346.
applications. As the embryo increases in size late in in- Islam, A. F., S. W. Walkden-Brown, C. W. Wong, P. J. Groves,
S. K. Burgess, K. E. Arzey, and P. L. Young., 2001. Influence
cubation, the likelihood of injecting the embryo proper of vaccine deposition site on post-vaccinal viraemia and vaccine
increases and, with correct injector-to-egg alignment, efficacy in broiler chickens following in ovo vaccination against
results in targeted delivery to the right breast area. Mareks disease. Avian Pathol. 30:525533.
Why the delivery results of the Intelliject system did Sarma, G., W. Greer, R. P. Gildersleeve, D. L. Murray, and A. M.
Miles. 1995. Field safety and efficacy of in ovo administration
not reflect this is unknown. of HVT + SB-1 bivalent Mareks disease vaccine in commercial
This study shows the large variability in accuracy of broilers. Avian Dis. 39:211217.
vaccine deposition, depending on the in ovo vaccination Sharma, J. M., L. F. Lee, and P. S. Wakenell. 1984. Comparative vi-
system used. Previous work done by Islam et al. (2001) ral, immunologic, and pathologic responses of chickens inoculated
with herpesvirus of turkeys as embryos or at hatch. Am. J. Vet.
and Wakenell et al. (2002) demonstrated that vaccine Res. 45:16191623.
protection against challenge was dependent on the vac- Sharma, J. M., and R. L. Witter. 1983. Embryo vaccination against
cine delivery site. The vaccine must be injected deep Mareks disease with serotypes 1, 2 and 3 vaccines administered
singly or in combination. Avian Dis. 27:453463.
enough into the egg to be delivered into the amnion, Wakenell, P. S., T. Bryan, J. Schaeffer, A. Avakian, C. Williams,
which includes the AM and EM. When delivered to and C. Whitfill. 2002. Effect of in ovo vaccine delivery route on
the AM, the vaccine is imbibed, breathed in, or both HVT/SB-1 efficacy and viremia. Avian Dis. 46:274280.
by the embryo before hatching. The MD vaccination Zhang, Y., and J. M. Sharma. 2001. Early posthatch protection
against Mareks disease in chickens vaccinated in ovo with a
delivered via the AL results only in significantly lower CVI988 serotype 1 vaccine. Avian Dis. 45:639645.
viremia and does not adequately protect against chal-