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avian insight
V.1 2003
avian insight
3/7/05
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avian insight
A L O H M A N N A N I M A L H E A LT H N E W S B R I E F
V. 1 2 0 0 3
Extracted from a presentation Evaluation of the effect of heating an oil emulsion Pasteurella multocida bacterin on tissue reaction and immunity. Karen E. Burns, Jaime Ruiz, and John R. Glisson. Poultry Diagnostic and Research Center, University of Georgia. Presented at AAAP, 2001 Boston.
Lesion in neck muscle of breeder pullet following improper injection technique of an oil emulsion vaccine
# of Birds
50
10 week
Vaccination - Right Serology
18 week
Vaccination Left Serology
24 week
Challenge & Serology (25 birds) Lesion Score (25)
In this issue of avian insight: Warming oil emulsion bacterins prior to injection . . . . . . . . . . . . . p.1
Room Temp Vaccine 50 Vaccination - Right Serology Vaccination Left Serology Challenge & Serology (25 birds) Lesion Score (25)
Controls
50
Serology Only
Serology Only
Right and Left indicate side of the breast muscle where injection occurred. Groups were divided at 24 weeks, 1/2 challenge and 1/2 lesion scored. continued on page 2
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2 continued from page 1 in a foam-insulated container for transport to pens for immediate injection. Room temperature vaccine was placed out of refrigeration for 12 hours and allowed to warm to room temperature. The birds were injected intramuscularly in the superficial pectoral muscle with 0.5 mL of the bacterin using an 18-gauge 1/4 inch needle. Antibodies to P. multocida were measured using the IDEXX ELISA for P. multocida. 25 birds from each treatment group were challenged at 24 weeks of age with 1.0 mL serotype 1 P. multocida (X-73 challenge strain, 2.0 X 102 colony forming unit/mL) by I.M. injection in the left thigh.
avian insight
V.1 2003
Lesion Scoring. 25 birds from each treatment group were euthanized at 24 weeks of age. Both breast muscles were carefully incised to reveal the superficial and deep pectoral muscles. The following scoring system was used to subjectively evaluate the injection lesions:
0 = no visible lesions
3 = large focal or multifocal lesions > 1cm in diameter, including lesions that extended into deeper muscle layer
1 = small focal lesion within one muscle layer, < 1cm in diameter 4 = abscessation, deep pectoral myopathy, lesions that were diffuse, > 5cm in diameter
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Results
Table 2 shows the ELISA data at 10, 18 and 24 weeks. Titers for P. multocida tend to be highly variable and will have higher CV% than typically seen with viral titers.
Discussion
The heating of a P. multocida bacterin to 41 C (105 F) was compared to an industry-accepted standard of room temperature, 25 C (77 F). Tissue reactivity, serological response, and
Table 2: P. multocida ELISA Geometric Mean Titers from trial 1 comparing treatment groups. Treatment Group Heated Room temp Control Week 10 5 16 38 CV% 181% 112% 74% Week 18 1635A 946B 15 CV% 66% 73% 137% Week 24 3136 2841 7 CV% 56% 82% 121%
Antibody response, as measured by ELISA, induced by the heated vaccine displayed a significant difference in GMT at 18 weeks. One possible explanation for this difference is a change in the absorption rate of the first vaccine, resulting in higher titers. The improvement in titer could be important in vaccine programs that provide one killed bacterin injection combined with a live wing-web injection. The difference in GMT between the two vaccinated groups did not remain significantly different at 24 weeks. Label recommendations for most manufacturers suggest warming oil emulsion vaccines to room temperature prior to injection. The results of this study indicate a potential advantage to warming to higher temperatures, anywhere in a range 85-105 F (29-41 C). The bacterin in this study was warmed to 105 F with some cooling allowed, with the estimated temperature of the product being 90-95 F. An accurate temperature of the injected product could not be obtained without sacrificing sterility of the bacterin. Since the study was limited to one product, further research, addressing duration of immunity, the persistence of lesions and emulsion stability should be completed to fully evaluate this technique. However, field application has proven successful in reducing local and systemic reaction from injection of bacterins. Additionally, vaccine crew members report a distinct advantage in the ease of administration when the vaccine is warmed to above room temperature. This study was conducted on a standard water-oil emulsion product. With new adjuvant and emulsion technologies available in commercial products, this technique may not show an advantage for those types of formulations. Despite this potential aid in reducing vaccine reaction, focus should still remain on proper injection technique and accuracy to reduce the potential harmful effects of vaccination.
AB significance at (P<0.05). For lesion scores, the general trend observed was that birds receiving heated vaccine displayed lower lesion scores as compared with room temperature with all controls displaying a lesion score of 0. There was a significant (P<0.05) difference between the number of birds scoring in lesion groups 2 and 3 when comparing the two treatments. More birds in the room temperature group had lesion scores greater than or equal to 3. Figures 1 and 2 depict the lesion scoring at 6 and 14 weeks post vaccination.
50
40
Heated
# of Birds 30
20
10
0
Score 0 Score 1 Score 2 Score 3 Score 4
50
40
80 70
30
60
% Mortality
20
50 40 30 20
10
0 10 0
Heated Vaccine
b a
RT Vaccine Controls
Treatment
Fig. 1. Lesion score distribution at 6 weeks post vaccination. Mortality caused by challenge demonstrated the non-vaccinated controls had a significantly higher (P<0.05) mortality, 76%, than the two vaccinated groups. When comparing the heated and room temperature group, the mortality rates were significantly different (P<0.10), 0% and 12% respectively, as seen in figure 3.
Fig. 3. Mortality following challenge with Serotype 1, 6 weeks post second vaccination. protection from challenge were all measured to examine differences in the treatment. The findings indicated that when a P. multocida bacterin is heated to 41 C prior to I.M. injection into the breast, a significant reduction occurred in tissue reactivity based on the lesion scoring system.