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avian insight

V.1 2003

from the president…
New product labels
As 2003 gets off to a start, Lohmann Animal Health International is launching a “new look” in the United States market with our newly designed LAH labels, which symbolize our company’s relationship with Lohmann Animal Health. We have nearly completed our transformation from the dual company that functioned as Vineland/MBL, and we have grown into our own as Lohmann Animal Health International. As a part of Lohmann Animal Health, we stand for quality products and services and a sincere dedication to the poultry industry. We have structured ourselves to be very lean and productive, exactly like our poultry production customers. The new labels carry our LAHI corporate name at the bottom, our Gainesville, Ga., headquarters address, and our recently approved single USDA Establishment Number. All of the information is framed by a colorful stripe down the left side of the label. Look for them when you get your next vaccine shipment. These new labels will now be submitted to international regulatory officials to update our product registrations in more than 60 countries. It will take more than a year to complete this phase of label introduction.

Inactivated vaccines are the core of our success
We are the global leader in inactivated vaccines. Our success lies in our unique adjuvant approaches, our individualized service approach with autogenous vaccines in the U.S. and in Germany, and our enhanced vaccine production capabilities with our state-of-theart, fully automated plant in Maine. As an example of this, the popularity of our broiler breeder vaccine, BTO2-REO, is growing in the U.S. broiler business. Its 100 percent bursa-derived IBD antigen is coupled with the industry leader in REO formulation. This vaccine is perfect to protect young broilers and allow them to produce up to their genetic capacity. Ask your LAHI area manager for a visit from our technical support group to program BTO2-REO into your operation. And, while you’re at it, include a trial with FC4 Gold™, the low dose, intramuscularly applied Fowl Cholera vaccine with a special adjuvant designed for 21st century breeders.

Production consolidation streamlines operations
We are in the final phase of consolidating all U.S. vaccine production to our recently modernized site in Maine. With production facilities in Maine and Cuxhaven, Germany, we can easily accommodate the growing global vaccine production needs for our company. The exception to this approach is the production of Marek’s vaccine, which, for the immediate future, will continue to be made at our New Jersey facility. We are in the final stages of completely revamping people, processes, facilities and materials to improve our ability in this major product area. We have run validation serials and are now entering the market with more consistent Marek’s vaccines. We intend to be a long-term supplier of the best Marek’s vaccines in the world, and our volume capabilities will continue to build through 2003.

avian insight

Gainesville, GA 30501 1146 Airport Parkway

For further information: 770.532.3627 • 800.655.1342 • www.lahinternational.com

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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

Warming oil emulsion bacterins prior to injection
Karen E. Burns, D.V.M, M.A.M, diplomate A.C.P.V Technical Services Veterinarian Lohmann Animal Health Gainesville, GA breast, thigh, or leg. However, there are inherent problems in all of the above vaccination techniques. For example, the S.Q. neck injection offers the highest risk for accidental human injection, a medical emergency. Commonly, a deeper injection into the muscles of the neck occurs, resulting in pain, feed refusal, and cull birds. Oil emulsion bacterins tend to be the most reactive when injected into the muscle. The tissue reaction can result in a greater incidence of condemnations at processing, further decreasing the low monetary return from spent hens. In order to examine a field solution to the problem of I.M. injection reaction, a simple investigation was completed at the University of Georgia to determine if heating the vaccine would reduce the reaction in the breast muscle. Personal communication from an industry vaccination crew indicated that they were heating an oil emulsion bacterin in a water bath prior to I.M. injection into the breast. It is known that heating an oil emulsion decreases the viscosity of the vaccine. In theory, by adjusting the vaccine closer to the birds’ body temperature, the vaccine may be less likely to leave a lesion at processing, possibly because of more rapid absorption of the vaccine. Comparing the heated vaccine to room temperature vaccine allowed evaluation of a commonly used field technique and label recommendation.

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.

Materials and Methods
Oil emulsion inactivated vaccines are critical components of vaccination programs for longlived birds, i.e. breeders, layers and turkeys. These vaccines provide a long duration of immunity and improve antibody response for breeders in order to provide protective maternal antibodies to progeny. Common injection sites include subcutaneously (S.Q.) in the back of the neck, ventral side of the tail, and intramuscularly (I.M.) in the The study used a commercially prepared P. multocida bacterin containing serotypes 1, 3, and 4 from the same serial and lot, refrigerated until use. The heat treatment was performed by placing the bottle in a microbiological incubator set to 41 C (105˚ F) for five hours before vaccination. The vaccine was placed

Lesion in neck muscle of breeder pullet following improper injection technique of an oil emulsion vaccine

Table 1: Study Organization Group
Heated 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

From the president . . . . . . . . . . . . . p.4

Challenge & Serology (25 birds) Lesion Score (25)

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

2 = small multifocal lesions of < 1cm in diameter, same muscle layer;

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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

Room Temp Control

20

10

0
Score 0 Score 1 Score 2 Score 3 Score 4

Breast Lesion Score
* Significance at P < 0.05

Fig. 2. Lesion Score Distribution at 14 weeks post vaccination.

50

40

80 70

c

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.

a,b,c indicates significance at P < 0.1

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.