FLEX Symposium & PCVAD Research Award Winners Presentation

FLEX Symposium & PCVAD Research Award Winners Presentation St. Paul, Minnesota Ingelvac CircoFLEX Ingelvac MycoFLEX Ingelvac CircoFLEX-MycoFLEX

Table of Contents FLEX Symposium September 20, 2008 Performance of conventional healthy pigs vaccinated with Ingelvac MycoFLEX or Suvaxyn RespiFend... 1 John Kolb, DVM; et al Impact of PCV2 Vaccination on Subclinical PCVAD Cases in Canada, France and USA...3 Ernest Sanford, DVM, Dip Path, Diplomate ACVP An innovative method for quantifying animal behavior responses to various immunization protocols...6 Keith Bretey, DVM; et al Ingelvac CircoFLEX - The Bleeding Edge of Innovation Toward the Control of Complex Disease Syndromes... 10 Mike Roof, M.S., Ph.D Ingelvac CircoFLEX Ingelvac MycoFLEX Ingelvac CircoFLEX-MycoFLEX

Performance of conventional healthy pigs vaccinated with Ingelvac MycoFLEX or Suvaxyn RespiFend J Kolb, D Johnson, E Diaz, R Edler Boehringer Ingelheim Vetmedica, Inc., Ames, Iowa, USA Introduction and Objectives This paper describes the results of a field evaluation of a novel M. hyo bacterin, Ingelvac MycoFLEX (Boehringer Ingelheim Vetmedica, Inc., St. Joseph, Mo.), and another commercially available vaccine in a conventional production system positive for PRRSv, Salmonella typhimurium, Lawsonia intracellularis and SIV and Mycoplasma hyopneumoniae (M. hyo). Materials and Methods A commercial production system was selected that utilized wean to finish production and vaccination against M. hyo at approximately three and five weeks of age. A total of 1254 pigs were weaned into the barn over a 10-day period to fill 46 of 48 pens. Two pens remained open for use as hospital pens. All pigs within a given pen received the same vaccination, with the exception of two non-vaccinated sentinel pigs per pen which were used for lung lesion and serologic evaluations. Vaccinated pigs received either Suvaxyn RespiFend (2 ml, two doses at 3 and 5 weeks of age) or Ingelvac MycoFLEX (one 1 ml dose at 5 weeks of age). The volumes given should be mentioned somewhere. Pigs were observed for any local or systemic side effects following injection, which had previously been commonly reported by the production system following vaccination with Suvaxyn RespiFend. Non-vaccinated sentinel pigs (92) were tagged with a purple tag, while vaccinated pigs were tagged with either a red tag (two pigs per pen for serologic evaluations) or yellow tag. All pigs had a unique and consecutively numbered tag regardless of tag color. All pigs were weighed off test over two days. A subset of each group of pigs was selected for slaughter check, with at least five pigs per pen eligible for examination. All selected pigs remaining on the farm at the time of the health check were examined. Pigs were randomly selected during off-test weighing and lung lesions were scored blindly by a veterinarian (73 non-vaccinated sentinels, 85 Suvaxyn RespiFend and 92 Ingelvac MycoFLEX pigs, respectively). Average daily gain was the performance parameter of interest in this evaluation and was analyzed using one-way ANCOVA, having treatment and sex as main effects and day 0 weight as a covariate. Individual pig was the experimental unit. Starting weight was evaluated using one-way ANOVA. LS means are reported. Results No significant difference (p>0.05) in average daily weight gain (ADG) was detected between the vaccinated groups. Both groups of vaccinated pigs had significantly reduced average lung lesion scores (ALLS) as compared to non-vaccinated pigs (p 0.05, Table 1). 1

Twenty-seven percent of pens had at least one pig that suffered adverse systemic side effects following injection with Suvaxyn RespiFend. Affected pigs exhibited seizures, salivation and lethargy. One pig died. No pigs exhibited any type of adverse reaction following Ingelvac MycoFLEX vaccination. The day following vaccination, more Ingelvac MycoFLEX pigs were observed at feeders than Suvaxyn RespiFend pigs (2.4 vs. 1.6 pigs/pen, respectively). Table 1. Summary Data (LS means) Non-vaccinated sentinels* Ingelvac MycoFLEX Suvaxyn Respifend Total Weight Gain, lbs NA* 218.08 a 216.90 a ADG, lbs NA* 1.46 1.46 Average Lung Lesions Score, % 23.3 a 5.3 b 8.6 b a,b: p<0.05. ADG=average daily gain. ALLS=average lung lesion score (% of lung with lesions). *Non-vaccinated sentinels used only for lung lesion & serologic evaluations. Conclusions Both vaccinated groups had similar average daily gains and end weights. Ingelvac MycoFLEX pigs exhibited fewer local and systemic side effects than pigs vaccinated with RespiFend. This provides an interesting contrast between the oldest, traditional two-dose M. hyo vaccine and a new, non-reactive single-dose vaccine. Excellent efficacy is obtainable with a safe, non-reactive, singledose vaccine. 2

Impact of PCV2 Vaccination on Subclinical PCVAD Cases in Canada, France and USA Ernest Sanford, DVM, Dip Path, Diplomate ACVP Boehringer Ingelheim Vetmedica (Canada) Tel: 519-641-1785; Fax: 519-641-2334 E-mail: ernest.sanford@boehringer-ingelheim.com Introduction Once PCV2 pig vaccines started to be widely used two years ago, it quickly became apparent that the biological performance of PCV2-vaccinated pigs in many cases exceeded historical levels observed in herds prior to their PCVAD outbreak. The implication of these observations was that the ubiquitous PCV2 infection in pigs, which most of us had passively accepted as being benign, might actually have been present at a subclinical level of intensity and constraining productivity to an extent not recognized prior to the availability of PCV2 pig vaccines. This paper details three studies that evaluate the impact of PCV2 vaccination on pigs subclinically affected by PCVAD. The studies were conducted on pigs in three different countries Canada, France and USA using a single dose of Ingelvac CircoFLEX vaccine (1-3). Canadian Study Pigs were sourced from a high health, PRRSV- and Mycoplasma hyopneumoniae-negative, PCV2-positive, 2,850-sow herd in which PCVAD had never been diagnosed clinically or histopathologically. Included in the trial were 1,056 pigs, 528 of which were injected with 1 ml of Ingelvac CircoFLEX and 528 injected with 1 ml of a placebo (sterile water) at 3 weeks of age. Pigs entered the finishing barn at 54 days of age (after 33 days in the nursery). Pens of pigs were weighed and feed intake measured biweekly upon entry into the finishing barn. The pen was the experimental unit. All pigs were weighed at days 2 and 88 (end of study) in the finisher. Carcass data were collected from the slaughter plant. The mixed model procedure of SAS statistical software was used to analyze the data. Results Vaccinated pigs were on average 2.5 kg (5.5 lb) heavier than unvaccinated controls (107.3 kg [236.1 lb] vs. 104.8 kg [230.6 lb] [p<0.01]), respectively, and had an ADG in finishing of +36 g (0.79 lb) compared with the controls (p<0.001). Mortality was greater (p<0.05) for the controls vs. vaccinates by 4.5 percent. Carcass weight, lean percent and index were greater (p<0.05) for the vaccinates. Loin depth was 1.8 mm greater for the vaccinates over the controls (P<0.05). 3

French Study Pigs were from a 900-sow, PRRSV- and M. hyopneumoniae-positive, farrow-to-finish herd. For the study, 1,548 pigs, 2-3 weeks of age, were assigned to vaccinated (Ingelvac CircoFLEX) or unvaccinated groups, commingled and individually weighed at 3, 10, 15, 20 and 23 weeks of age. Viremia was determined by qpcr on sera from 250 pigs sampled every other week up to 22 weeks of age. Body weights were compared using ANOVA and subsequent t-tests and the percentage of viremic animals, clinical signs and mortality were compared using Fischer s exact test. The individual pig was the experimental unit. At the time of the study the herd was categorized as a subclinical PCVAD herd as clinical signs of PCVAD were not observed. Results At the end of the trial, vaccinated pigs were on average 2.4 kg (5.28 lb) heavier than unvaccinated controls (p 0.0001). ADG was 27 g (0.59 lb.) higher for the vaccinates compared to the controls (p<0.0001) during the grow-finish period (10 to 22 weeks of age). General health status was good for both groups. At peak viremia (17 weeks of age) 9.4 percent of the vaccinates and 55 percent of the unvaccinated controls were viremic (p 0.0001). Mortality in finishing (15 to 22 weeks of age) showed a trend toward vaccinates over the controls (3.02% vs. 1.59%) but was not statistically significant (p=0.0859). USA Study Pigs were from a PRRSV-negative but M. hyopneumoniae-positive, multi-site production system. The study consisted of 2,400 pigs (1,200 vaccinated with Ingelvac CircoFLEX and 1,200 unvaccinated controls), with the pen being the experimental unit. Treatment groups were alternated by pen. Each barn had 40 pens housing 30 pigs per pen. Pen weights were collected at 3, 12 and 22 weeks of age. Blood samples were collected serially from 10 randomly selected pigs from each group at 3, 6, 10, 18 and 22 weeks of age. PCVAD lesions had been identified histologically in individual pigs in this herd but clinical signs of PCVAD had not been observed, hence its designation as a subclinical PCVAD herd. Results Both treatment groups were considered clinically normal throughout the trial although PCVAD was confirmed histologically and by qpcr in individual pigs at 10, 14 and 17 weeks of age, thus confirming subclinical PCVAD. ADG for vaccinated pigs in the grow-finish stage (12 to 22 weeks of age) was 930 g (2.05 lb) vs. 900 g (1.98 lb) for the unvaccinated controls (p=0.01). Viral loads of PCV2 were statistically significantly lower for vaccinates vs. controls (p<0.0001) on qpcr at 10, 18 and 22 weeks of age. Summary and Concluding Remarks The common thread running through all three studies is that vaccinated pigs consistently outperformed unvaccinated cohorts for a wide range of production parameters in herds that had historically not experienced clinical PCVAD. For nearly every parameter in all three studies the vaccinates had a statistically significant advantage over the unvaccinated pigs in these subclinical 4

PCVAD herds. From these and other studies (1-5) it is clear that PCV2 infection in growing pigs, in the absence of any observed clinical signs, can have a significant negative impact on performance. Since nearly all pigs in virtually all herds worldwide become infected with PCV2 some time during their growing period, it is possible that, unbeknownst to us in the past, overall productivity is being compromised in what we previously considered normal or even superior growing pigs. References 1. Young M, Cunningham G, Sanford E. Performance of Ingelvac CircoFLEX vaccinated pigs in a subclinical PCVAD herd. IPVS 20: Vol 1, pg 25, 2008. 2. Orveillon F-X, Fachinger V, Denotte J-P. Effect of PCV2 vaccination on growth performance in a subclinically affected herd. A.D. Leman Swine Conf 35: In Press. 3. Bretey K, Diaz E. Efficacy of Ingelvac CircoFLEX in a herd with subclinical PCVAD. A.D. Leman Swine Conf 35: In Press. 4. King D, Dubois P, Painter T, et al. Biological and economic benefits of controlling subclinical PCVAD with PCV2 vaccination. IPVS 20: Vol 2, pg 36, 2008. 5. Henry S, Dritz S, Nietfeld J, et al. PCV2 studies: Research from the K-State PCV2 team. AASV 39: 463-467, 2008. 5

An innovative method for quantifying animal behavior responses to various immunization protocols Keith Bretey, Roy Edler, Edgar Diaz Introduction Pigs are naturally curious about their environment and seek out any recent changes or introductions into that environment. Pigs also are cautious, initially acting fearful or excitable when a person first enters a pen. However, after a 15-second adjustment period, pigs that have no previous reason to fear people will relax and explore the presence of a person in the pen by nosing or biting at the observer s legs and feet. The National Pork Board s Swine Welfare Assurance Program 1 (SWAP) introduced in 2003 utilized these behavioral principles to identify and quantify pigs that are relaxed in the presence of people in a pen. After entering a pen and kneeling down in that pen, 50 percent of the pigs in that pen should return to the observer s gloved hand or approach the observer and/ or demonstrate a relaxed posture within the 15-second observation period. An adaptation of the animal observation section of the 2003 SWAP has been employed and suggests that willingness to approach may be a useful parameter for assessing vaccine reactivity (side effects) in pigs 2. A similar adaptation of the animal observation section of the 2003 SWAP was utilized in this study to evaluate the behavior of weaned pigs following vaccination with Intervet/Schering-Plough s Circumvent /M+PAC ½ dose compared to Ingelvac MycoFLEX, Ingelvac CircoFLEX, a combination of both products, or saline as measured by clinical observations pre-injection and six hours post-injection. Circumvent and Ingelvac CircoFLEX are both vaccines to aid in the reduction of circovirus disease, while M+PAC and Ingelvac MycoFLEX are both vaccines to aid in the reduction of disease caused by Mycoplasma hyopneumoniae. The Circumvent/ M+PAC protocol used in this study was chosen because it was the normal protocol used in the production system and because it is a protocol reportedly used by veterinarians in some other swine production systems. This protocol is not used at label dosage in this evaluation and the products are not approved for mixing/combined use. Per the system s normal regimen, a second dose of Circumvent/M+PAC was given 2 weeks later, however, the behavioral response to the second dose was not quantified. The terms safety, reactivity and approachability as used in this document refer to relative responses by groups of pigs to various injectable vaccines or saline, as measured by the relative decrease in percent of pigs in a group that are willing to approach an observer within a specified time interval, with the baseline value for each group determined prior to treatment and the same measurement repeated six hours post-treatment. Objectives To evaluate the comparative reactivity of Ingelvac CircoFLEX alone, Ingelvac MycoFLEX alone, Ingelvac CircoFLEX/Ingelvac MycoFLEX mixture, Circumvent/M+PAC mixture and saline injection as measured by clinical observations pre-injection and six hours post-injection. 6

Material and Methods On day 0 (the day before immunization) an observer blinded to treatment entered each of the study pens and recorded the number of pigs that approached the observer within a 15-second period of time. The 15 seconds of observation time began after the observer entered the pen, sat down on a stool, and tentatively extended a leather-gloved hand in the direction of the majority of the pigs. The observer then recorded the number of pigs that approached the observer in that 15-second period of time. A pig was considered to have approached the observer if that pig demonstrated a non-threatened stance and both eyes of the pig could be seen by the observer. On day 1 the immunization products or sterile saline were administered to approximately 6,250 pigs in 250 pens from four barns (two rooms per barn) beginning at 7:20 a.m. and utilizing multiple vaccination teams. All immunizations were given in the muscle on the right side of the neck. Beginning at 1:20 p.m. that same day (six hours post-injection) and progressing at the same rate as the vaccination crew, an observer blinded to treatment entered each of the study pens in the same order as the immunizations occurred and recorded the number of pigs that approached the observer within a 15-second period of time. The 15 seconds of observation time began after the observer entered the pen, sat down on a stool, and tentatively extended a leather-gloved hand in the direction of the majority of the pigs. The observer then recorded the number of pigs that approached the observer in that 15-second period of time. A pig was considered to have approached the observer if that pig demonstrated a non-threatened stance and both eyes of the pig could be seen by the observer. Each population of pigs served as its own baseline for behavior observations. The treatment groups were identified as follows: 1] Saline (2 ml) 2] Ingelvac CircoFLEX/Ingelvac MycoFLEX mixture (2 ml) 3] Ingelvac CircoFLEX (1 ml) 4] Ingelvac MycoFLEX (1 ml) 5] Circumvent/M+PAC ½ dose mixture (2 ml) Randomization procedures: 1) Baseline data (percentage of the pigs that approached) from each pen was sorted by room within barn from lowest to highest. 2) A random number was assigned to each pen. 3) Within blocks of five pens, treatment groups 1-5 were assigned to pen based on lowest to highest random number. Results Baseline pre-injection There were no differences in the pre-treatment (baseline) percentage of pigs per pen that approached the observer among the five treatments (Table 1). 7

Six-hour post-injection There were no differences in the percentage of pigs per pen that approached the observer between saline, Ingelvac CircoFLEX and the mixture of Ingelvac CircoFLEX and Ingelvac MycoFLEX. The percentage of pigs approaching the observer was lower in the Ingelvac MycoFLEX group compared to the saline group (p 0.05). The percentage of pigs approaching the observer was significantly lower in the Circumvent/M+PAC ½ dose group compared to saline, Ingelvac CircoFLEX, Ingelvac MycoFLEX as well as the mixture of Ingelvac CircoFLEX and Ingelvac MycoFLEX (Table 1). Change from baseline to six-hour post-injection behavior (delta) There were no differences in the change (delta) from baseline to six-hour post-injection percentage between saline, Ingelvac CircoFLEX or the mixture of Ingelvac CircoFLEX and Ingelvac MycoFLEX. There was a significant change from baseline to six hours post-injection behavior for the Ingelvac MycoFLEX group. However, this difference was still significantly smaller than the change observed with the mixture of Circumvent/M+PAC ½ dose. All four treatments had significantly lower changes from baseline approachability (delta) compared to the Circumvent/M+PAC ½ dose group (Table 1). Table 1. Willingness to approach observation results Item Saline CircoFLEX (CF) MycoFLEX (MF) CircoFLEX + MycoFLEX (CF/MF) Circumvent + M+Pac (CV/MP) Number of pens observed 54 51 49 50 46 Number of pigs observed 1350 1275 1225 1250 1150 Baseline pre-injection behavior, % interested Six hour post-injection behavior, % interested Change from baseline to six hour post injection behavior, % difference 64.89 64.08 63.51 63.04 62.43 55.33 a 54.59 ab 51.35 b 51.52 ab 37.13 c 9.56 a 9.49 a 12.16 b 11.52 ab 25.30 c abc Within a row without a common superscript differ (Fischer s Exact Test P 0.05). 8

Conclusions Approachability decreased in pigs that received Circumvent/M+PAC ½ dose compared to saline, Ingelvac MycoFLEX, Ingelvac CircoFLEX or both products. This method of observing pigs and measuring percent approachability was repeated in two other projects with similar results. These findings suggest that: 1) Willingness to approach is a useful parameter for assessing vaccine reactivity (side effects) in pigs, 2) Ingelvac CircoFLEX and the mixture of Ingelvac CircoFLEX and Ingelvac MycoFLEX were no more reactive than saline and Ingelvac MycoFLEX was only slightly different from saline, and 3) The Intervet/Schering-Plough product mixture tested was significantly more reactive than saline and all tested BI products. References 1. National Pork Board. Swine Welfare Assurance Program 2003. Animal Observation: 12-17. National Pork Board, Des Moines, Iowa, USA. 2. Baumert, D. et al. Willingness to approach behavior and feed disappearence of weaned pigs following vaccination with Mycoplasma vaccines. 20th IPVS Congress, P02.030, Durban, South Africa, June 2008. Ingelvac, CircoFLEX and MycoFLEX are registered trademarks of Boehringer Ingelheim Vetmedica, GmbH, Ingelheim, Germany. Circumvent is a registered trademark of Intervet, Inc., Millsboro, Delaware. M+PAC is a registered trademark of Schering-Plough Animal Health Corporation. 9

7 Mike Roof Ingelvac CircoFLEX The bleeding edge of innovation toward the control of complex disease syndromes PCVD - Global perspective and successful control, Durban, June 24 th, 2008 47 10

Mike Roof PCVD - Global perspective and successful control, Durban, June 24 th 2008 Boehringer Ingelheim Vetmedica, Inc., Ames, Iowa USA Ingelvac CircoFLEX The bleeding edge of innovation toward the control of complex disease syndromes The global swine industry has undergone incredible change and development over the past decade. Changes in herd size, management practices, and emerging diseases have all re-shaped the industry. Associated with these changes, animal health companies also need to change and adapt to insure they provide tools that meet the current industry needs as well as those of the future. Historically, animal health companies focused on vaccine registration with goals related to addressing a single clinical issue. However, with the emergence PRDC (Porcine Respiratory Disease Complex) and PCVAD (Porcine Circovirus Associated Diseases) we have moved away from dealing with clinical disease associated with a single etiological agent and move into complex disease syndromes involving multiple pathogens that interact in a complex fashion and wreak economic havoc for swine producers. In the late 1990 s Boehringer Ingelheim Vetmedica recognized that Porcine Circovirus type II (PCV 2) was an important and emerging disease. Because of our commitment to the swine industry we made this pathogen a priority in our R&D efforts. However we quickly determined that control of PCV 2 alone would provide some value to the industry, but that long term we needed to develop FLEXIBLE tools, tools that worked globally, tools that took into account regional needs, and tools that addressed complex syndromes. The cornerstone of our effort was built and designed around our Ingelvac CircoFLEX vaccine, containing the unique combination of Purified Circovirus Antigen (PCA TM ) and the highly effective aqueous polymer adjuvant ImpranFLEX TM. In starting this project, we worked with Boehringer Ingelheim scientists, veterinarians, and global disease experts. We felt in order to develop a world class vaccine, we needed to know and understand the following key issues: 1. Where does PCVAD disease most commonly manifest itself within the global swine population? 11

2. If a vaccine were available, when/where would it be used to be most effective? 3. What would be the properties of a world class vaccine that would provide the most benefit to swine producers across the globe? 4. How to best match customer demands for efficacy, safety and convenience all at the same time? of PCV 2, PRRSV, and M. hyopneumoniae as the most commonly detected agents in this disease syndrome. Therefore, we concluded that ultimately, not only would we need to develop a tool to control PCV 2, but it must be compatible with other vaccines based on the regional needs and disease agents. It needed to be FLEXIBLE to the herd and customer. When we looked at herds across the globe we routinely found that almost all pigs and pig herds were PCV 2 positive for antibody and antigen and yet not all herds had clinical disease. When we focused only on herds that had clinical disease we consistently noted that the timing of the disease was almost exclusively in the grower and finisher (> 6 weeks of age). This suggested that pigs in their first week of life, although exposed to the PCV 2 virus were likely protected by maternal immunity. Disease was then observed after maternal decay. Based on this assessment, sow vaccination and inducing additional maternal immunity didn t make sense or seem to add value to a control program. Sows were already positive and maternal immunity already appeared to be present and yet pigs were experiencing PCVAD. Vaccination of a sow seemed like it would have little benefit and may only push the disease into older and more valuable finisher animals. This was the basis for our focus on a pig vaccine and not on a sow vaccine. In hindsight this was a great decision! Numerous studies and reports have been done looking at PCVAD and investigating the presence of swine pathogens. In general almost all reports, no matter on the method or geographical region, consistently list some combination So at this point, we knew we had a disease that was found in the grower and finisher. We knew we needed to vaccinate pigs and induce immunity in the pig. We needed a vaccine that could be used with other respiratory vaccines. So the next step was to consider WHEN to vaccinate a pig for optimal protection. Because there were many reports of clinical disease in the grower (> 6 weeks of age), we knew that an effective vaccine had to placed and used prior to this time and yet be given enough time to induce protective immunity PRIOR to the clinical disease. So we worked backwards. > 6 weeks of age Clinical Disease Observed 3-4 weeks General time to induce a maximum immune response 3 weeks of age Optimal time to vaccinate pigs and insure disease control!! We knew that ORF 2 was a neutralizing epitope and so this would be the basis of a quality PCV 2 vaccine. Vaccination at 3 weeks of age would mean that a 2 dose vaccine would not be of value and potentially induce immunity too late. We didn t have time to get 2 doses into pigs before disease started and would leave herds susceptible during early infections and maternal decay. PCVD - Global perspective and successful control, Durban, June 24 th, 2008 49 12

Traditional grow it, kill it, 2 dose vaccine technology is easy and commonly used in autogenous vaccines and older commercial products that are used in naive animals. However development of 1 dose vaccine technology and vaccines that work in the face of maternal antibodies requires: A high quality, purified, stable, neutralizing antigen, at the proper dosage PCA TM (Purified Circovirus Antigen). The PCA TM is forming a hollow PCV-2 virus, so live virus like antigen is presented to the immune system. A world class adjuvant that induces rapid onset of immunity, has good duration of immunity, and induces the proper immune response ImpranFLEX TM. ImpranFLEX TM is a buffered solution containing an aqueous polymer. It has an excellent safety profile as it does not contain mineral oil. A high quality scientific team A little dose of LUCK! The conclusion of 7 years of research and development by the BIVI R&D team was the US launch of Ingelvac CircoFLEX in 2006 and the European launch in 2008. Ingelvac CircoFLEX is a one-shot vaccine, 1ml per dose, with an outstanding safety profile. It has been proven to override maternal immunity in pigs from 3 weeks of age onwards. The vaccine provides a rapid onset of immunity as early as two weeks after vaccination and has proven to deliver protective immunity through to slaughter in close to 100 million pigs. The product has taken the market by storm and consistently exceeded expectations in PCV 2 and PCVAD disease control. In former times this would have been enough and the end of the story However, as was stated earlier we no longer work in a world where control of a single etiological agent is enough. We need to provide solutions that meet complex disease syndromes. For this reason we have developed the Ingelvac CircoFLEX in parallel to another new vaccine Ingelvac MycoFLEX. This new vaccine was launched in the US in 2008 and registration in other countries is in progress. This will be the second phase of our delivery of the FLEX concept to the swine industry. 1. Ingelvac CircoFLEX vaccine 1 ml/1 dose 2. Ingelvac MycoFLEX vaccine 1 ml/1 dose 3. Formulated with the same adjuvant, ImpranFLEX. 4. Ingelvac MycoFLEX CircoFLEX-MycoFLEX and Ingelvac combination CircoFLEX has can been mixed licensed, and allows which the allows use its of contents a 2 ml/ 1 Ingelvac CircoFLEX and Ingelvac MycoFLEX vaccines * Highly effective to be mixed in controlling and administered both as a single-dose, Mycoplasma 2 ml lung injection lesions to and pigs PCVAD 3 weeks of age at or the older. same time. * Convenient Highly effective dose in size controlling (1+1=2 ml) both Convenient Mycoplasma and lung cost lesions effective and PCVAD 1 dose program at the same time. Very Convenient safe with dose no size systemic (1+1=2 and ml) injection site reactions Convenient and cost effective 1 dose program 5. The Very data safe demonstrating with no systemic efficacy and of injection the FLEX site combination reactions was presented at the AASV San Diego in March 2008. 5. The data demonstrating efficacy of the FLEX combination was presented at the AASV San Diego in March 2008. 50 13

Treatment group Number of Animals Average lung lesion score (% of lung) Ingelvac MycoFLEX 18 5.5 a MycoFLEX mixed with CircoFLEX 19 3.9 a Table 1: Efficacy of Ingelvac MycoFLEX in FLEX Concept Study Challenge Controls 19 14.3 b Strict Controls 6 0.0 a,b: p < 0.0001 Treatment group Number of Animals Lymphoid depletion (%) Lymphoid inflammation (%) Lymphoid IHC (% affected) Ingelvac MycoFLEX mixed with 24 CDCD pigs 0a 4.2 a 8.3 a Ingelvac CircoFLEX Non-Vaccinated Controls 24 CDCD pigs 83.3 b 87.5 b 91.7 b a,b: Values with different superscripts differ significantly (p < 0.0001) Table 2: Efficacy of Ingelvac CircoFLEX in FLEX Concept Study We are truly excited about these innovative developments and hope that Boehringer Ingelheim give you the power to FLEX your muscle and control these economically significant disease complexes, raise healthy pigs, and maximize your economic returns. PCVD - Global perspective and successful control, Durban, June 24 th, 2008 51 14

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innovative solutions in swine health Boehringer Ingelheim Vetmedica, Inc. 2621 North Belt Highway St. Joseph, Missouri 64506-2002 1.800.325.9167 www.bi-vetmedica.com 2008 Boehringer Ingelheim Vetmedica, Inc. Ingelvac, Ingelvac MycoFLEX and Ingelvac CircoFLEX are registered trademarks and CircoFLEX-MycoFLEX is a trademark of Boehringer Ingelheim Vetmedica, GmbH, Germany. Always read, understand and follow the label directions.