avianinsight

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 August 2008

Volume 1

Live attenuated vaccines: benets of water stabilizers in drinking water and spray administrations
We will continue to invest in research, development and production to assure a steady supply of high quality avian biologics for the poultry industry.
Dave Zacek President, Lohmann Animal Health
By Ivn R. Alvarado, DVM, MS, Ph.D., ACPV, Technical Services Veterinarian, Lohmann Animal Health International

techniques for the administration of live attenuated vaccines. The stimulation of immunity in a massively vaccinated population (herd immunity) reduces the

A poor administration is still the most common cause of vaccine failure in poultry. A successful administration of live attenuated vaccines by spray or drinking

the ease and low cost have made spray and drinking water the preferred methods for massive administration of live attenuated vaccines in the farms.
probability of an individual bird in a vaccinated ock to become infected (Marangon and Busani 2006). Since under eld conditions, it is unrealistic to expect a 100% protection of the birds in a vaccinated ock, the goal is to develop immunity in a high enough proportion of birds to prevent the transmission or minimize the adverse effects of a disease agent. For poultry, the ease and low cost have made spray and drinking water the preferred methods for massive administration of live attenuated vaccines in the farms. water requires an adequate preservation and proper delivery of the vaccine to the host. In this article, we describe the adverse effects a live vaccine might encounter during resuspension and administration in suboptimal water, and the benets of using water stabilizers during vaccination.

Introduction
The increasing demand for economical and rapidly available sources of animal protein has highly inuenced the constant growth of poultry operations around the world. The capacity of poultry to adapt to diverse geographical conditions and the fact that no large areas of land are required, have also contributed to the expansion and efciency of commercial poultry operations. As a result of their size increase, it has become necessary to develop and implement massive

Massive Administration of Live Vaccines

During manufacturing, most live attenuated vaccines are subjected to a lyophilization process. Lyophilization, in

inside

Live attenuated vaccines p.1

Presidents Note

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association with adequate refrigeration temperatures (between 4C and 7C or 39F and 44F), guarantees the preservation of an adequate vaccine titer able to stimulate a satisfactory immune response in the bird. However, once reconstituted, the limited lifespan of live attenuated vaccines should be a major concern. Drinking water is an appropriate method of administration for live attenuated vaccines that require an initial infection within the gut, such as Salmonella, infectious bursal disease and avian encephalomyelitis vaccines. Also, due to the communication of the nasal cavity with the mouth via the choanal cleft, a successful application of respiratory virus vaccines (such as infectious bronchitis and Newcastle disease virus) should be expected. Coarse spray is a widely used and effective method to vaccinate birds against respiratory viruses, not only at the hatchery but also in the farm. By coarse spray, the vaccine is introduced through the eye, the nostrils and by ingestion after pecking.

Periodic treatments of water wells with high concentrations of chlorine release products (CRP), and chlorination of the drinking water through in-line proportioners, have been effectively used in poultry operations to control microbial levels. However, CRP also inactivate live vaccines. Chlorinated compounds (such as sodium hypochlorite, chlorine dioxide and sodium dichloroisocyanurate) are highly active oxidizing agents, destroying the cellular activity of proteins (McDonnell and Russell 1999). Sodium hypochlorite ionizes to produce Na+ and the hypochlorite ion OCL-, which is transformed in hypochlorous acid (HOCL), the active moiety responsible for bacterial inactivation (McDonnell and Russell 1999). The deleterious effects of CRP on bacteria include a disruption of oxidative phosphorylation and other membrane-associated activity as well as inhibition of DNA synthesis. Hypochlorous acid levels as low as 2.6 ppm completely inhibit the growth of E. coli within 5 minutes, 96% of DNA synthesis and between 10% and 30% of protein synthesis (McDonnell and Russell 1999). The virucidal activity of CRP includes nucleic acid fragmentation. pH is a measure of acidity or basicity of a solution. pH measurements depend on the concentration of hydrogen (H+) and hydroxide ions (OH-). Waters with neutral pH (ph = 7.0), have an equal concentration of H+ and OH-. The resuspension of live vaccines in low pH water will cause structural damage to cell membranes and macromolecules such as DNA and proteins. Week acids, such as some organic acids used in drinking water for poultry, can pass freely

through the bacterial cell membrane. Once inside, the week acids dissociate and increase the acidication of the cytoplasm, affecting the bacterial survival in drinking water (Cotter and Hill 2003). Furthermore, a synergistic effect between low water pH and the bacteriocidal/virucidal activity of chlorine-released agents has been demonstrated. While low pH values (below 5) adversely affect water consumption, high pH values can affect water palatability and form mineral deposits which reduce the water ow rate (Vermeulen, Backer et al. 2002). Hardness is a measure of the quantity of salts, mainly of divalent ions such as calcium, magnesium and/or iron, in the water. Hardness is expressed as milligrams of calcium carbonate (CaCO3) per liter (ppm) and indicates the total quantity of divalent salts present (Wurts 1993). However, high hardness in the presence of low calcium but high magnesium levels can be observed. A previous research study at Lohmann Animal Health has shown the detrimental effect of hard water (20 ppm Fe and 4 ppm NaOCl) on Newcastle disease virus and Salmonella typhimurium live vaccines two and four hours after resuspension. Drastic reductions in viral and bacterial titers have been observed 2 and 4 hours after resuspension in hard water. In areas with hard water, the constant deposit of insoluble minerals in the nipple drinkers and water lines can also affect the homogeneous administration of live vaccines to the ocks. Spray vaccination using hard water can create mineral deposits, clogging the spray nozzles.

Adverse Conditions
At the farm, a poor condition of the water used during resuspension and administration will have a detrimental effect on vaccine titers. Live vaccines should be resuspended in fresh water with a close to neutral pH, moderate hardness and no chlorine or other disinfectant residues. The undesirable effects these factors will have on the survival of live vaccines are briey described in the following paragraph.

Water stabilizers
Live poultry vaccines are produced in specic pathogen free (SPF) embrionated eggs, tissue cultures or nutritional broths under controlled laboratory conditions. Vaccine manufacturers have recognized the adverse conditions live vaccines face during their administration in the farm. For such reason, the use of water stabilizers during vaccination is recommended worldwide by technical services veterinarians.

additive. Other reducing agents are sodium metabisulte, sodium bisulte and ammonium thiosulfate (Simpson 2001). pH buffering agents: used to stabilize the water pH between 6 and 7, approximately. Some buffering agents include sodium phosphate, potassium phosphate, sodium citrate, sodium bicarbonate and potassium bicarbonate. Water soluble dye: food colored dyes

and administer the vaccines. Furthermore, the inclusion of dyes facilitates the visualization of vaccine in water lines and vaccinated birds. The use of water stabilizers during vaccination allows a proper preservation of live vaccines, which, in association with a proper administration technique, will contribute to a successful vaccination.

References
Barbour, E. K., A. Abdelnour, et al. (2002). Evaluation of 12 stabilizers in a developed attenuated Salmonella Enteritidis vaccine. Vaccine 20 (17-18): 2249-2253. Cargill, P. (1999). Vaccine administration in poultry. In Practice 323-328. Cotter, P. D. and C. Hill (2003). Surviving the acid test: responses of Gram-positive bacteria to low pH. Microbiol. Mol. Biol. Rev. 67(3): 429-453. Marangon, S. and L. Busani (2006). The use of vaccination in poultry production. Rev. Sci. Tech. Off. Int. Epiz 26(1): 165-274. McDonnell, G. and D. Russell (1999). Antiseptics and disinfectants: activity, action and resistance. Clinical Microbiology Reviews: 147-149. Simpson, G. D. (2001). The reduction of the Chlorite ion. International symposium on chlorine dioxide., Las Vegas, Nevada. Vermeulen, B., P. D. Backer, et al. (2002). Drug administration to poultry. Adv. Drug Del. Rev. 54: 795-803. Wurts, W. A. (1993). Understanding water hardness. World Aquaculture 24(1): 18.

One of the rst products used to stabilize the drinking water during vaccination was skimmed milk. Dilution of skimmed milk (approximately 2 g/liter or 10 g/gallon) in the drinking water at least 20 minutes before the addition of the vaccine was a general recommendation (Cargill 1999). Skimmed milk has shown to efciently overcome the detrimental effects of chlorine. However, the presence of undissolved residues in cold water, which can eventually block nipple drinkers, and the presence of residues inside the water pipes after vaccination, serving as a source of nutrients for bacterial growth (build up of biolms), has limited its use. More efcacious dyed water stabilizers for the protection of live vaccines are commercially available. These are some of the characteristics and components used in water stabilizers: Reducing agents: used to neutralize oxidizing sanitizers (i.e. residual chlorine) or contaminants present in farm water. One of the most commonly used reducing agents is sodium thiosulfate (Simpson 2001), which is recognized as a safe food

approved by the Food and Drug Administration are essential to allow the detection of the water containing the live vaccines in the drinker systems and in vaccinated birds (tongue and nasal cleft after eye drop administration; tongue, oral cavity or crop after water administration, and skin and feathers after coarse spray vaccination). Thermo-stability conferring agents: bovine serum albumin, sorbitol, maltose, lactose, sucrose and glycerol are some of the most common vaccine stabilizers (Barbour, Abdelnour et al. 2002).

Conclusions
The administration of live attenuated vaccines by massive methods such as drinking water and spray is a very common practice in poultry operations. Massive administration methods are not as efcient as individual vaccination. However, they offer several advantages, such as low cost and rapid administration of one or several vaccines. Since live attenuated vaccines are very fragile organisms, it is important to neutralize the adverse chemical agents present in the water used to resuspend

Presidents Note
hired additional staff are now fully integrated and part of the vaccine manufacturing team in Maine. The majority of our production consists of inactivated vaccines. Increased demand for AviPro vaccines both in the USA and internationally,
Dave Zacek President, Lohmann Animal Health

be completed by fall of 2009. In addition, we are adding a fermentation suite in the manufacturing plant. This new production addition will allow increased volumes of our bacterial products, both live and killed. Recognition of AviPro quality is driving the demand. We are pleased that customers value our vaccines and keep demanding more year after year. We will continue to invest in research, development and production to assure a steady supply of high quality avian biologics for the poultry industry. And we will continue to work hard to be your Avian Professionals.

require we build again. We have begun a large, state-of-the-art animal testing complex on property adjacent to our current manufacturing plant in Winslow. Included on the site are the Animal Services Administration Center, a large pen isolation unit, small test houses and a large multiple isolation unit. Ground was broken for phase I of this project June 2008. The multi-million dollar facility will

It has been nearly one year since we opened our newly expanded production/testing/label, pack and ship site in Maine after having closed our Vineland site and transferred vaccine production to Winslow, Maine. Newly

375 China Road Winslow, Maine 04901

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207.873.3989 800.655.1342 www.lahinternational.com