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from the president

Dave Zacek, President of Lohmann Animal Health International, Gainesville, Georgia, USA

In addition, we are using a proprietary adjuvant described as water-in-oil-inwater (WOW). This new adjuvant is designed to provide long duration of protection without causing harsh postvaccination reactions. AviPro 101 Coryza Gold is our way of answering customer demand for improved efficacy and improved duration of immunity against this disease. Practical Development of New Vaccines Lohmann Animal Health International has produced a steady stream of new avian vaccines since our inception in 2000. Each vaccine was developed in response to specific customer needs. Our approach has been practical and

close to the user. Look to us for a continuous supply of new avian vaccines. Our company is focused on only avian vaccines. This allows us to devote a significant part of our revenues to developing new vaccines. And, to learn how to use AviPro 101 Coryza Gold in your operation, contact your LAHI Area Manager or LAHI Veterinarian for more information.

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A L O H M A N N A N I M A L H E A LT H I N T E R N AT I O N A L N E W S B R I E F V. 1 2 0 0 5

EFFICACY AND DURATION OF IMMUNITY STUDIES INVOLVING A HAEMOPHILUS PARAGALLINARUM BACTERIN IN A NOVEL OIL EMULSION
Karen Burns GroganA, Patricia S. WakenellB, Ken TakeshitaC, Eric J. LovellD
Technical Service Veterinarian, Lohmann Animal Health International, Gainesville, GA Associate Professor, Population Health and Reproduction, College of Veterinary Medicine, University of California at Davis, Davis, CA. C Area Manager/Technical Service Veterinarian, Lohmann Animal Health International, Vacaville, CA D Consultant Veterinarian, Lohmann Animal Health International, Pensacola, FL
A B

Introducing a New Vaccine We are very pleased to introduce AviPro 101 Coryza Gold, a new vaccine for protection against Coryza (H. paragallinarum). Our new vaccine contains representative strains of type A, B and C H. paragallinarum to provide broad protection against a significant variety of expected challenges.

Lohmann Animal Health International is a leading manufacturer and supplier of poultry biologics for the U.S. and international vaccine markets. Based in Gainesville, GA, LAHI maintains production facilities in Vineland, NJ, and Waterville, ME. LAHI is a member of the PHW Group, a 32-company enterprise that produces avian vaccines and poultry for the global market.

Introduction Haemophilus paragallinarum (H. paragallinarum) is the etiology of Infectious Coryza, a highly contagious, acute respiratory disease of poultry. This disease can be economically devastating in tropical and subtropical parts of the world, where it is more commonly reported. Clinical signs of the disease include nasal exudates with a characteristic odor, sinusitis, conjunctivitis, with depression and lethargy. Infected laying flocks can experience a reduction in egg production by 40-100% and will typically recover with treatment. Even with the use of commercially available bacterins, affected flocks can experience production drops of 5-10%. H. paragallinarum isolates were originally grouped into serovars by Page in 1962 using a plate agglutination test. Isolates were placed into one of three serovars, A, B, or C. In 1983, another identification scheme was reported, the Kume scheme, which grouped isolates using a hemagglutination inhibition test. The Kume scheme was then adapted
In this issue of avian insight:

Nasal exudates seen in infectious coryza

Characteristic sinusitis present in infectious coryza

to correlate with the existing Page serovars. Additionally, more recent work has reported a hemagglutination inhibition test for the Page scheme, resulting in the most accepted identification system for serovar identification. Serovar distinction remains critical to discussions of bacterins since only homologous protection is known to exist, meaning inactivated antigens of group A will only protect to group A. There is no cross-group protection. Cross protection is reported to exist among isolates within serovars. The bacterins produced against H. paragallinarum are reported to exhibit adverse post vaccination reactions, especially those in water-in-oil emulsions. These reactions can result in decreased feed consumption, a delay in sexual maturity, and a reduction in total number of eggs produced per hen. In general, the bacterins that are water-

based in aluminum hydroxide offer less post vaccination reaction, but protection can be reduced. The ideal vaccine would include all three serovars incorporated into an adjuvant that delivered the antigens to the immune system stimulating a long-lived, high level of immunity without adverse reactions. To minimize the field problems with coryza bacterins, Lohmann Animal Health International (LAHI) has developed and licensed a new commercial H. paragallinarum bacterin, presented in a novel oil emulsion. The product, Coryza ABC Gold, contains highly antigenic quantities of representative strains from the three Page serovars of H. paragallinarum. In order to evaluate the bacterin in the novel adjuvant, the following studies were performed to demonstrate the reduction in tissue reaction, protection from challenge and duration of immunity.

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Gainesville, GA 30501 1146 Airport Parkway

Efficacy and Duration of Immunity Studies Involving a Haemophilus Paragallinarum Bacterin in a Novel Oil Emulsion ..............p.1 From the President ...............p.4

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For further information: 770.532.3627 800.655.1342 www.lahinternational.com

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Study 1 Evaluation of Tissue Reactivity

Table 1. Scoring system to evaluate SQ injections
Score Description of Lesion 0 11 1+ 2 3 4 No visible lesion No adjuvant remnants with slight inflammation; or traces of emulsion with no inflammation Remnants of adjuvant with only minor inflammation Remnants of adjuvant with some edema and/or petechial hemorrhage deeper in the tissue Single or multiple well-defined granulomas, less then 1 inch in size Single or multiple well-defined granulomas, 1 inch in size or greater Larger granulomas and/or diffused swelling of the subcutaneous or muscle tissue with necrotic lesions

product in the novel emulsion. One hundred fifty SPF chickens, five weeks of age, were divided into six groups. Three groups were vaccinated subcutaneously in the neck with one dose (0.5 ml) of the experimental bacterin and revaccinated four weeks after the initial vaccination. The remaining three groups served as non-vaccinated, chalGraph 1: Percent protection by group following challenge

in commercial laying flocks. Birds were injected twice with the LAHI coryza bacterin, subcutaneously at ten weeks and 14 weeks of age. The flocks injected with Coryza ABC Gold were compared to flocks injected with another commercially available bacterin. The parameters evaluated were tissue reaction by manual palpation at two weeks post vaccination, mortality following injection, and production effects. Graph 2 will display the tissue reaction results from one participating farm. In the farms participating, no significant differences were observed in mortality rates, egg production and other production parameters (feed conversion, weight gain, etc.) between groups vaccinated with the LAHI product as compared to the other commercial bacterin. University Study: Duration of Immunity To evaluate the efficacy of the product in field-vaccinated birds, a university collaborated with LAHI to complete a challenge trial in commercial layers from the previous field studies. Eighty field-vaccinated birds at 48 weeks of age, that had received two doses of LAHI Coryza ABC Gold, and 40 age-matched non-vaccinated control birds were transported to university challenge facilities. After an acclimation
Strain Name Serogroup A - 0083 Serogroup B - SpRoss Serogroup C - Modesto CFU per dose 8.3 x 106 6.8 x 105 3.4 x 107

Table 3: Results following challenge with serotypes A, B, C in 48-week-old commercial layers.

Group 0083 Vaccinated Non-vaccinated # Positive % Protection # Positive % Protection
a,b -

Challenge SpRoss 1/20a 95% 7/10b 30% Modesto 0/20a 100% 8/10b 20% 0/20a-100% 8/10b 20%

indicate significance at P< 0.01 by Chi Square test

Graph 3: Percent protection following challenge with three strains of H. paragallinarum.

This bacterin using a novel adjuvant containing the three serovars of H. paragallinarum was tested against a popular water-based commercial bacterin also containing all three serovars. Previous field experience with this commercial water-based product has demonstrated the desirable minimum post vaccination reaction. Groups of threeweek-old specific pathogen free (SPF) birds were vaccinated subcutaneously with either the experimental vaccine or the commercial water-based vaccine. Three weeks post vaccination, the vaccination sites of the birds were evaluated for adverse reaction and scored numerically with a standardized lesion scoring system. The lesion scoring system appears in Table 1. The experimental vaccine produced an average injection site reaction score of 0.63 compared to the commercial water-based vaccines higher score of 1.9.
Table 2. Percent protection following intranasal challenge
Group Vaccinated Non-Vaccinated Vaccinated Non-Vaccinated Vaccinated Non-Vaccinated Serovar Challenge A (8.46 x 10 CFU)
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lenged controls. At two weeks post revaccination all birds were challenged intranasally with one of the three challenge strains of H. paragallinarum. The challenge dose varied by serovar, see Table 2 for actual challenge levels. The chickens were observed daily for signs of facial swelling or nasal exudates for a period of ten days post-challenge. All birds were housed under conditions of high temperature and high humidity throughout the observation period. Chickens exhibiting two consecutive days of facial swelling and/or two consecutive days of nasal exudates during the observation period were considered positive for H. paragallinarum infection. Results are found in Table 2 and in Graph 1. Field Studies As a part of the licensing procedure, the product was tested
Graph 2: Lesion Score results from field-vaccinated flocks

% Protection 100% 28.6% 91.7% 20.0% 93.8% 5.6% Lesion scoring system for this trial:
0 = No palpable lesion 1 = Palpable mass less than 1cm in diameter 2 = Palpable mass greater than 1 cm, but less than 2cm in diameter 3 = Palpable mass greater than 2 cm in diameter

period, birds were challenged intranasally with 0.30 ml of one of three challenge strains present in the bacterin, at the following titer levels: For the challenge period, birds were housed under conditions of high temperature and high humidity. Challenge groups were observed for eight days postchallenge for clinical signs of coryza facial swelling or nasal exudates. Any bird displaying clinical signs for two consecutive days during the eight-day period was judged as positive for H. paragallinarum infection. Results of challenge and percent protection can be found in Table 3 and Graph 3.

Conclusions Through the studies presented, this new H. paragallinarum bacterin has been shown to be highly effective. The product produces immunity to all three serovars contained, showing equal protection to challenge in both studies presented. University-performed challenge studies demonstrated effective immunity in birds at 48 weeks of age. Furthermore, in both in-house studies and field studies, this new bacterin proved to be less tissue reactive, even compared to water-based products. Developments in emulsion technology have allowed vaccine manufacturers to overcome previous negative effects experienced in the field with certain products; coryza bacterins are one such example. By developing an H. paragallinarum bacterin in a novel oil emulsion, Coryza ABC Gold from LAHI will alleviate previous problems experienced with this group of products. Further field studies are planned to extend the evaluation of this product, including the protective effect to egg production drops in the face of field challenge and use in broiler breeders.

A B (5.64 x 105 CFU) B C (8.64 x 107 CFU) C

Study 2 Bacterin Efficacy A challenge trial was completed to prove efficacy of the

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