Final Report Summary - THRIVE-RITE (Natural Compounds to enhance Productivity, Quality and Health in Intensive Farming Systems.) Executive Summary:Section 1: Executive Summary THRIVE-RITE is a collaborative project funded by the EU Seventh Framework Programme (Ref: 315198, start date: 01/08/2012, end date: 28/11/2014). The aims of THRIVE-RITE are to identify viable solutions to problems encountered in intensive production of monogastric animals. These problems include: rising food costs, lower producer margins due to lag effect (increase in input without increase at retail level), ban on in-feed antibiotics, reduced productivity and the limited effectiveness of current 'Solutions' to these problems. The use of in-feed growth promoting antibiotic use is no longer a tool legally available to producers to enhance productivity and control diseases on farms (Regulations (EC) No. 1831/2003 and 1334/2003). These are also human implications to these issues. While the ban on in feed antibiotics was intended to reduce the use of antibiotics in production of pig and poultry meat, this has not happened and in fact, the situation has worsened. Renewed concerns were raised in 2013 by the European Medicines Agency (EMA) regarding the increasing antibiotic use in the EU, an article which appeared in Der Spiegel - (http://www.spiegel.de/international/europe/danish-pig-farmers-reduce-antibiotics-to-prevent-drug-resistance-a-933344.html). The statistics demonstrate that huge levels of antibiotics are being administered by veterinarians in the EU. Germany is the worst offender and there are now major concerns of an imminent rise of drug-resistant pathogens in animal production. The main reason for increased antibiotics use is the lack of viable alternatives to in feed antibiotics following their ban in 2006. Zinc oxide (ZnO) is a widely used alternative. However, it faces increased scrutiny as the majority of the 3Kg of ZnO added per tonne of feed, is passed on through the faeces and into the environment.THRIVE-RITE was established to provide solutions to these problems. THRIVE-RITE consists of highly progressive European enterprises with established records in developing natural feed additives. The enterprises have developed novel, natural-based products which enhance animal health, performance and immunity, as demonstrated in numerous peer-reviewed publications. THRIVE-RITE combines the expertise of the enterprises with a multidisciplinary team of scientists in some of Europe’s leading research institutes. The combination of enterprise and academia provides a robust system in which to verify the use of these products in enhancing animal growth, health and performance, a requirement increasingly being set in modern agriculture. A dual approach was applied to validate the effectiveness of the products. This involved a 2-Stage validation process in (a) the academic setting and (b) large-scale commercial setting. This ensured that significant findings from the university setting were subject to a 2nd round of validation under large-scale commercial conditions. This provided a unique mechanism in which to transfer scientific discovery within academia to direct provision of solutions which are proven to be effective in commercial-scale production. The main targets achieved by THRIVE-RITE are:• Enhanced productivity as measured by ADG & FCR -Target achieved• Reductions in pig and poultry mortality - Target achieved• Reductions in levels of pathogenic bacteria and viruses - Target achieved• Increased antibody transfer to offspring via maternal transfer - Target achieved• Improved antioxidant content in pig & poultry meat - Target achieved• Enhanced gut health & peripheral immunity - Target achievedThe findings of THRIVE-RITE are described in this document, paying close attention to the project objectives and requirements of the end user to effectively enhance productivity and output, but in a manner which does not impact on public health or the environment.Project Context and Objectives:The THRIVE-RITE Consortium was established with the aims of developing effective and commercially viable solutions to problems facing food production in the EU. These problems include rising food costs and lower producer margins due to lag effects, the EU ban on in feed antibiotics and reduced productivity. With these challenges in mind, the THRIVE-RITE Consortium developed a plan focused on solving health problems in monogastric animals. THRIVE-RITE merged the technological and innovative prowess of SMEs with the knowledge and technical capacity of recognised experts in academia. The plan was developed according to the 4 principles listed hereunder & described in detail throughout this section.(a) Problems: Identifying the most pertinent problems facing EU agriculture.(b) Solutions & Targets: Identifying viable solutions to counteracting these problems.(c) Solutions Providers: Identifying solutions providers (SMEs) with a proven track record of innovation and generation of highly effective technologies.(d) Scientific Validation: for solutions to be proven, they must be validated at the highest of levels, including in academic and production settings.Problems facing EU agriculture:• Rising food costs- Population growth: The world’s population will expand to ~ 10 billion by 2050. Standards of living have increased dramatically, especially in China and India. This has increased demand for dairy-based products which has increased prices with knock on effects on feed prices. - Fuel costs: Fuel costs have increased rapidly over the last 10 years. This has directly impacted on feed prices as well as harvesting costs.- Weather extremes: In recent years, colder & wetter summers have been observed in NW Europe with drier weather in the south. This has dramatically affected production. The USA drought in 2012 led to cereal shortages and higher prices.• Lower producer margins due to lag effect: The biggest risk for producers is where input prices rise rapidly without corresponding increases at the retail level. This lag effect causes huge problems for producers.• Ban on antibiotics: In-feed growth promoting antibiotic use is no longer a tool legally available to producers to enhance productivity and control diseases on farms (Regulations (EC) No. 1831/2003 & 1334/2003). Salmonella & Enterobacteriaceae remain common disease burdens on farms and increasingly require direct treatment with antibiotics by veterinarians, with retailers also suffering from reputational damage. • Reduced productivity: Animals fail to thrive when infected by pathogens. In pigs, Enterobacteriaceae reduces performance and viruses such as Porcine circovirus type 2 (PCV-2) and Porcine reproductive and respiratory syndrome virus (PRRSV) impact on productivity, thus requiring vaccination. Important diseases in poultry include Enterobacteriaceae, Coccidial oocysts, Clostridium, Reovirus, Avian RhinoTracheitis, Infectious Bursal Disease, Chicken anaemia virus & Ornithobacterium rhinotracheale.• Current 'Solutions' to farming problems are limited: In-feed antibiotics increase growth & feed efficiency. Since their ban, alternatives comparable in effect have not been found. Use of ZnO in animal feed has also been banned in some EU counties. There remains no viable solutions to these problems.(b) Solutions and TargetsThe Consortium identified and targeted potential solutions to problems facing monogastric animal production, including: (a) products developed to achieve the stated objectives and/or (b) technological approaches to administering the product to obtain maximum efficacy. Specific targets for these technologies were set as follows:• Enhance productivity as measured by ADG & FCRProducts must demonstrate significant improvements in performance in pig and poultry animal models, as measured by enhancements in average daily gain (ADG) and feed conversion ratio (FCR). Enhancements of ADG and FCR by 11 and 7% respectively were targeted, i.e. levels previously achieved with in-feed antibiotics.• Reduce mortalityBenefits from products are likely to enhance overall herd/flock health status, an effect which should lower the incidence of mortality-related losses on farms.• Reduce levels of pathogenic bacteria and viruses A target was set to achieve a significant reduction in levels of pathogenic bacteria and virus load. Important infectious agents include Salmonella, Enterobacteriaceae, Campylobacter, Clostridium, Porcine Circovirus Type 2 (PCV-2) virus, Porcine Parvovirus (PPV) and Reovirus. The target was to reduce pathogen by 1 log.• Increase antibody transfer to offspring via maternal transfer:In addition to direct application, it was decided to assess efficacy in the offspring of treated animals. If effective, mechanisms such as 'maternal transfer' would offer a novel, cost-effective means of enhancing herd health and performance. • Increase protection of embryo by in ovo application of bioactiveDirect application of products in ovo can offer an effective means of enhancing chicken health and productivity. The target was to validate this technology in the academic and commercial scale, in order to develop a means of increasing chicken productivity, while at the same time, lowering the incidence of infectious diseases such as Campylobacter, a leading source of food-borne illness.• Improve antioxidant content in pig & poultry meatThere is evidence from some studies that application of products to animal diets can increase the levels of antioxidant in meat. The target in THRIVE-RITE was to improve meat quality and potentially shelf-life for pig and poultry producers.• Enhance gut health & peripheral immunityProducts in THRIVE-RITE have previously been shown to improve gut health and immunity while also modulating the peripheral immune response. Pig and poultry producers urgently require products which can achieve these effects on a large commercial scale. The target in THRIVE-RITE was to improve these key parameters.(c) Solution ProvidersAt the core of THRIVE-RITE are enterprises with the technological capacity to produce innovative products in an effective and commercially viable manner. Three enterprises in THRIVE-RITE have developed products which enhance animal health, performance and immunity, as demonstrated in numerous peer-reviewed publications. The products contain high concentrations of bioactive compounds, extracted from natural resources. Each group formulated their products towards maximum efficacy in different trials.(d) Scientific ValidationCritical to the success of THRIVE-RITE is independent scientific validation of product efficacy. For THRIVE-RITE to identify solutions to agricultural problems, products must be validated in both academic and large-scale commercial settings. This approach is highly rigorous. Recognised experts were selected to achieve the project targets in the academic setting (Validation Stage 1). In addition, large-scale commercial trials were undertaken in pig and poultry production settings to validate how products performed when beyond the university setting (Validation Stage 2).Validation Stage 1: AcademiaFour academic institutions tested the efficacy of these products under challenge and non-challenge situations. Their collective expertise are: virology, microbiology, immunology, in ovo technology and meat quality. They worked independently covering both academic and commercial validation of products. Products were tested on research farms by the academic groups as follows:Agri-Food and Biosciences Institute, Belfast (UK)- Parameters tested: Performance, PCV-2 virus, gut health & peripheral immunity- Animal models: Pig and Poultry- Application: Direct application, transfer to offspring via maternal transfer & in ovoUniversity College Dublin (Ireland)- Parameters tested: ADG, FCR, Salmonella, Enterobacteriaceae, Campylobacter, gut health & immunity- Animal models: Pig and Poultry- Application: Direct application & transfer to offspring via maternal transferUTP University of Science and Technology (Poland) - Parameters tested: ADG, FCR, Salmonella, Enterobacteriaceae, Campylobacter, gut health & immunity- Animal models: Poultry- Application: In ovo technology and application via waterUniversity of Molise in Campobasso (Italy) - Parameters tested: Meat quality- Animal models: Poultry meat - Application: In ovo technology and application via waterValidation Stage 2: Commercial-scale Two commercial enterprises have developed ultra-modern animal production facilities. This allowed for large-scale validation of findings, by demonstrating effects in a commercial setting. These conditions represented the normal commercial farm setting with increased risk of bacterial infections and viruses. Several tests were carried out at large scale by Drobex- Agro Sp. Z O.O. (Poland) and Truly Irish Country Foods Ltd. (Ireland). Trials were designed and implemented under the supervision of University College Dublin (Ireland) and UTP University of Science and Technology in Bydgoszcz (Poland). Project Results:Section 3: Description of main scientific and technical results/foregrounds 3.1 Project overview 3.1.1 Introduction THRIVE-RITE has 3 main work-packages (WP), each with specifically defined targets. A short summary of the findings from each data WP is outlined in Section 3.1.2 below and in greater detail in Section 3.2. The focus of each WP was as follows:• WP2: Enhancing animal health in the face of pathogenic challenge.• WP3: Maternal and direct applications for enhancing performance and meat quality • WP4: In ovo technology to enhance chicken health, performance and meat quality3.1.2 Project Summary (a) WP2: Enhancing animal health in the face of pathogenic challenge• Results: The aim of WP2 was to enhance animal health and performance in the face of challenge with Salmonella (Task 2.1 Task 2.3) PCV-2 virus (Task 2.2) and Campylobacter (Task 2.4). In Task 2.3 NeoShield® significantly increased ADG and reduced Salmonella typhimurium in experimentally challenged finisher pigs. FCR was improved by both NeoShield® and Bi2tos® in this trial. Colonic expression of immune genes IL-6 and TNF-alpha was significantly reduced by both NeoShield® and Bi2tos®. Increases in Lactobacilli spp. and intestinal expression of IL-10 and MUCB were also observed with Bi2tos®. Overall, this trial demonstrated the efficacy of prebiotics in enhancing health and performance in the important finisher life-stage of the pig. In the weaned pig, reductions in faecal score (Task 2.1) and reductions in PPV viral load in sera (Task 2.2) were achieved with NeoShield® in pigs experimentally challenged with PCV-2. In Task 2.4 NeoShield® demonstrated increased ADG and slaughter weight in broilers challenged with Campylobacter. • Conclusions: NeoShield® has been shown to be an effective product in enhancing performance in finisher pigs challenged by Salmonella typhimurium, while DiNovo® is effective in enhancing performance in chickens infected with Campylobacter. NeoShield® provided benefits to the weaned pig in terms of reduction in Porcine Parvovirus (PPV) viral load and improvements in faecal consistency. Both NeoShield® and Bi2tos® products modulated expression of several important immune genes, thus suggesting modes of action involving the immune response to infection, immune cell activation and inflammatory processes. Bi2tos® also influenced expression of genes which protect the gastrointestinal mucous barrier. (b) WP3: Maternal and direct applications for enhancing performance and meat quality. • Results: The aim of WP3 was to develop maternal and direct applications for enhancing weaned pig health, and to develop means of improving meat quality and shelf life. In Task 3.1 maternal sow feeding with LactoShield® was shown to improve faecal scores pre- & post-challenge with Enterotoxigenic Escherichia coli (ETEC). The dually treated LactoShield-NeoShield® groups had significantly decreased numbers of ETEC-LT versus the widely used commercial standard, zinc oxide (ZnO). Pigs from LactoShield® supplemented sows had growth performance equivalent to pigs offered ZnO during the post-weaning period. In Task 3.2 (experiment 1), LactoShield treated animals showed higher levels of IgG, IgA and IgM than untreated animals at the two earliest critical time points (6 and 11 days of age). In addition, LactoShield increased ADG significantly at several time points (between days 0-7, 25-32, 40-46, 53-60 and 25-78) compared to untreated animals (experiment 1). In Task 3.2A (experiment 2), LactoShield® and NeoShield® were shown to differentially enhance immune response and performance in pigs challenged with PCV-2 & PRRS virus. LactoShield treated pigs showed a strong PCV-2 specific antibody response with levels similar to PCV-2 vaccine alone, with significantly higher levels when compared to non-treated pigs on day 35, 42 and 49 post-infection. A joint university and commercial trial was held in Truly Irish Country Foods Ltd. and UCD facilities in Task 3.3. In this trial, LactoShield® significantly enhanced the life time growth performance of pigs up to slaughter age (Exp.1 UCD). LactoShield® had beneficial effects in the suckling period and improved faecal consistency during the post-weaning period (Exp. 2, Truly Irish Country foods). A low energy diet + NeoShield® significantly improved performance of pigs through improving FCR (Exp. 3). In Task 3.4 NeoShield® significantly improved pork meat quality, antioxidant content and shelf-life, and in many cases performed better than the commercial standard, Vitamin E. A number of significant benefits were also observed with Bi2tos®, including lower TVCs at several time-points and increased antioxidant content at the final day of storage, also greater than Vitamin E. • Conclusions: WP3 demonstrates the efficacy of LactoShield® and NeoShield® in reducing pathogens and improving weaned pig performance. This may offer a means of replacing the usage of Zinc oxide in the post weaning stage of pig production. WP3 also demonstrates that LactoShield® conferred health benefits to weaned pigs when compared to untreated animals. In the large-scale trial, LactoShield® was also shown to enhance the life time growth performance of pigs up to slaughter age. WP3 showed that NeoShield® is as effective and in some cases, more effective than vitamin E in improving meat quality, antioxidant content and shelf-life. (c) WP4: In ovo technology to enhance chicken health, performance and meat quality• Results: In this work package, the use of “in ovo” technology in broiler production was examined. In ovo technology involved optimizing the systems for injecting of the prebiotics into the egg, in ovo (Task 4.1 and Task 4.2). The delivery system was optimized further in Task 4.3 by examining the impact of in ovo application of each product on performance, microbiological and metabolic measures. Significant increases in broiler weight were achieved when DiNovo®, Bi2tos® and Lupin RFOs were administered via in ovo or via water alone, along with improvements in meat quality (Task 4.3). Both DiNovo® and Bi2tos® significantly increased levels of beneficial bacteria (Lactobacillaceae) during the 3rd week of chicken growth. Assessment of meat quality traits demonstrated that each prebiotic significantly improved meat colour (reduced redness), increased breast muscle fiber diameter and increased pectoral muscle weight. The ratio of polyunsaturated fatty acids (PUFA) to saturated fatty acids (SFA) was significantly increased in the DiNovo® group compared with the control. There was a trend towards increased intramuscular fat infiltration in the LAC group versus control (P=0.07) thus potentially improving meat flavour, “juiciness” and tenderness. In the commercial trial in Task 4.4 significant reductions in Clostridium and coccidial oocysts were achieved by all products in the commercial trial, with Bi2tos® also reducing Enterobacteriaceae (D4.4). Antibody responses specific to 5 pathogens were also significantly modulated by products, including antibodies against Avian RhinoTracheitis (ART), Infectious Bursal Disease (IBD), Chicken anaemia virus (CAV), Ornithobacterium rhinotracheale (ORT) and Avian infectious bronchitis virus (IBV). Intestinal morphology was differentially altered by each prebiotic. Prebiotics influenced meat quality, with delays in progression of oxidation observed with Lupin RFO. In the face of Reovirus challenge (D4.5) in ovo injection of DiNovo® significantly reduced chicken mortality, reduced viral load in the liver and increased chicken weight.• Conclusions: WP4 demonstrated differential effects of the products in enhancing broiler performance via in ovo administration and administration via drinking water. In Task 4.3 significant improvements in performance and microbiota were observed for each prebiotic. Substantial improvements in meat quality were also achieved. In the large-scale commercial trial (Task 4.4) the in ovo system was successfully implemented. Products were observed to have differential effects in modulating the immune system, reducing pathogenic infection and enhancing meat quality. In ovo injection of DiNovo® was shown to be an effective treatment for enhancing growth and survival of chickens infected with Reovirus. 3.2 Results 3.2.1 Introduction The ultimate measure of success of the THRIVE-RITE Consortium is the extent to which the problems identified have been solved. The primary aims of THRIVE-RITE were to develop products and application technologies which could achieve the targets listed below. Each of the main targets were met in THRIVE-RITE. These findings are described in detail throughout this section. A summary table is also provided below which outlines the significant results from THRIVE-RITE with respect to the main targets of the project.• Enhance productivity as measured by ADG & FCR• Reduce mortality• Reduce levels of pathogenic bacteria and viruses • Increase antibody transfer to offspring via maternal transfer• Increase protection of embryo by in ovo application of bioactive• Improve antioxidant content in pig & poultry meat• Enhance gut health & peripheral immunity3.2.2. Enhanced productivity as measured by ADG & FCRA central aim of THRIVE-RITE was to develop means of effectively enhancing productivity as measured by ADG, FCR and weight gain. These targets were successfully achieved in both pigs and chickens. Moreover, improved performance was increased in the face of challenge by problematic farm pathogens such as Salmonella, PCV-2 virus, PRRSV, Reovirus. The key findings are summarised as follows:(a) Chickens challenged with Campylobacter jejuni (Task 2.4)The efficacy of prebiotics in enhancing broiler performance in the face of Campylobacter challenge was assessed. Significant increases in ADG and slaughter weight were achieved on treatment with DiNovo®. (b) Pigs at first week post-weaning (Task 3.1) There was a significant dietary treatment effect on ADG of pigs pre-challenge (P<0.05). ADG for all LactoShield® treatments were statistically indistinguishable to that achieved with the commercial standard, ZnO. (c) Pigs challenged with PCV-2 and PRRSV virus (Task 2.2/3.2) The effects of LactoShield® and NeoShield® on the performance of weaned piglets challenged with PCV2 and PRRSV viruses were assessed. Unvaccinated pigs reared on LactoShield® treated sows and then treated with NeoShield®, had significantly increased ADG compared to the untreated unvaccinated control (days 27-33). LactoShield® and NeoShield® significantly increased ADG between Days 80-87 compared to the untreated vaccinated and unvaccinated groups. Vaccination alone did not significantly increase ADG versus untreated, unvaccinated control. Overall, this trial demonstrates the LactoShield® significantly increases performance in PCV-2 & PRRSV viral challenged pigs and to levels comparable to and at one time point greater than the vaccinated pigs (day 80-87).(d) In ovo administration of prebiotics (Task 4.3 4.4)The effects of prebiotics on enhancing chicken body weight were examined in a small scale-trial in Task 4.3. This trial focused on comparing the efficacy of prebiotic administration via in ovo injection or through administration via water alone. Each prebiotic were shown to significantly increase body weight of chicken at various time-points compared to the untreated control, with Lupin RFOs being the best. In the large-scale commercial trial in Task 4.4 trends towards increased chicken body weight were observed through in ovo injection of Bi2tos® and DiNovo®, the latter also showing substantial improvements in FCR. DiNovo® was also associated with improvements in European Broiler Index (EBI), thus suggesting benefits to the end user.(e) Broilers challenged with Reovirus (T4.5)This task evaluated the effects of in ovo inoculation with prebiotics on resistance to an enteric virus challenge (Reovirus challenge model). Significantly higher chicken body weight was achieved on the day of hatching for the DiNovo® treated group versus the uninfected control. Body weights at the end of the experiment were significantly higher with DiNovo® compared to the infected control, thus demonstrating the efficacy of DiNovo® in enhancing performance in the face of viral challenge.3.3.3. Reduced mortality and/or increase protection of embryo An important aim of THRIVE-RITE was to develop means of effectively reducing the incidence of mortality in pig and poultry production. In a Reovirus challenge trial in chickens, treatment with DiNovo® was shown to significantly reduce the mortality rates of infected chickens compared to the untreated infected control (1.74% versus 8.75%, p-value = 0.0131). In a large-scale commercial pig production trial (Task 3.3) maternal supplementation with LactoShield® was associated with a significant reduction in the average number of stillborn pigs (0.43 vs 0.68; p=0.05). In a PCV2/porcine parvovirus (PPV) dual infection, colostrum-deprived piglet infection model (D2.2) mortality was reduced in the NeoShield® group compared to the untreated group, statistical significance (5% vs 20%, p-value>0.05). In other trials, mortality levels were generally low.3.3.4 Reduced levels of pathogenic bacteria and viruses Animals fail to thrive when infected by pathogens such as Enterobacteriaceae and Salmonella. Viral agents also impact on productivity, thus requiring vaccination programs in many instances. A key aim of THRIVE-RITE therefore, was to develop a means of effectively counteracting pathogenic infection. The key findings are summarised as follows:(a) Salmonella Typhimurium in Finisher Pigs (Task 2.3)NeoShield® significantly reduced levels of Salmonella Typhimurium in faeces 7 days post infection (DPI) in finisher pigs. Levels of Salmonella were also significantly reduced by almost 1 log in the colon, with reductions also observed in the caecum. This trial demonstrates the efficacy of NeoShield® in reducing Salmonella levels in finisher pigs.(b) Enterotoxigenic Escherichia coli (ETEC) in weaned pig (Task 3.1)A combined treatment of LactoShield® and NeoShield® resulted in a significant decrease in levels of ETEC-LT, compared to the untreated control in weaned pigs. In contrast, treatment with zinc oxide, a commonly used commercial alternative to antibiotics, was associated with a significant elevation in the levels of ETEC-LT. While this trial demonstrates that combined treatment of LactoShield® and NeoShield® achieved significant reductions in ETEC-LT, it also demonstrates a limitation to the commercial standard, zinc oxide.(c) Pathogenic infection in chickens (Task 4.4)A significant reduction in Clostridium levels (~2.5 log) were achieved with each prebiotic on day 21 of chicken growth. In addition, Bi2tos® significantly reduced Enterobacteriaceae by ~ 3 logs on day 42. Coccidial oocysts were also significantly reduced by all prebiotics on day 42. DiNovo® showed the most significant (P≤0.01) anti-coccidial action in the beginning of the growing period (day 21), with reductions also reaching significance on day 42. Lupin RFO showed the strongest reduction on day 42. Overall, this trial demonstrates significant differential effects of each prebiotic in reducing levels of pathogens of relevance in poultry production.(d) Reovirus infection in chickens (D4.5)This task evaluated the effects of in ovo inoculation with prebiotics on effective resistance to an enteric virus challenge (reovirus challenge model). In ovo injection of DiNovo® was associated with signficant reductions in Reovirus viral load in the liver at 6 days post-infection. In addition to reduced viral load, DiNovo® was also associated with significant increases in chicken body weight and reductions in post hatch chicken mortalty, thus demonstrating the efficacy of DiNovo® in the face of Reovirus challenge.(e) Porcine Parvovirus Infection (PPV) in weaned pigs (D2.2)Treatment with NeoShield® was associated with significant reductions in serum viral load 4 days post infection in weaned pigs, as determined by quantitative PCR. This trend was observed to continue for the remainder of the trial.3.3.5 Increased antibody transfer to offspring via maternal transferOne mechanism of improving the health of young animals is to increase antibody transfer to offspring via maternal transfer. A key target of the THRIVE-RITE project was to increase levels of serum IgG, IgA and IgM in the weaned pig. (a) Pigs challenged with PCV-2 and PRRSV virus (Task 2.2/3.2 and 3.3) In Task 3.2 (experiment 1), a significant increase in colostral IgG, IgA and IgM concentrations was observed. In addition, a tendency for increased colostral IgG concentrations in commercial sows was also observed in Task 3.3 (experiment 1). Dietary treatment of sows with LactoShield® significantly increased IgA, IgG and IgM in the serum of progeny compared to untreated groups vaccinated against PCV-2 and those unvaccinated (Task 3.2). Moreover, these effects were observed within the first 60 days of the piglet’s life during which time the animal is under considerable stress. The NeoShield® group was associated with significant increases in immunoglobulins later in life. These findings demonstrate significant increases in immunoglobulin transfer from sow to piglet on treatment with LactoShield®, in a PCV-2 and PRRSV viral challenge situation.3.3.6 Enhance gut health & peripheral immunityAnimal health and productivity can be enhanced by mechanisms which improve gut health and peripheral immunity. Important aspects of gut physiology and immunity were targeted for improvements in THRIVE-RITE, the results of which are outlined below.(a) Finisher pigs challenged with Salmonella Typhimurium (Task 2.3)Both NeoShield® and Bi2tos® significantly reduced colonic expression of IL-6, a cytokine expressed by immune cells which play key roles in controlling acute phase response to infection, immune cell activation, and inflammatory processes. NeoShield® and Bi2tos® also significantly reduced colonic expression of Tumor necrosis factor alpha (TNFα), a pro-inflammatory cytokine secreted by innate and adaptive immune cells and stimulates the acute phase reaction and response to infection. Ileal expression of an anti-inflammatory cytokine, IL-10, was increased with Bi2tos®. Increased expression of MUC2B, a gene encoding a secretory protein with important roles in maintaining the gastrointestinal mucous barrier, was observed in the ileum of pigs treated with Bi2tos®. Modulation of the immune system in this manner is likely to reflect, in part, the underlying mechanisms which contributed to the significant reductions in Salmonella Typhimurium and increases in performance observed in the NeoShield® group, and some of the trends observed with Bi2tos®.(b) Pigs challenged with ETEC (Task 3.1)This trial compared the potential immune-modulatory effects of LactoShield® and NeoShield® with ZnO. Piglets from sows supplemented with LactoShield® and those also receiving NeoShield® exhibited increased expression of IL-8 in the ileum when compared with the untreated and ZnO treated groups. In addition, the LactoShield® treatment group had increased ileal expression of TFF3 but reduced colonic expression of TFF3 compared to the untreated group. Both LactoShield® and ZnO were associated with decreased expression of IL-6 in the colon when compared with the untreated group. The similarities and differences observed between treatments in modulating the immune system of the piglet may explain in part, the differential effects observed in enhancing performance and reducing ETEC-LT expression. In particular, LactoShield® was associated with performance levels comparable to ZnO while the LactoShield-NeoShield® combination significantly reduced ETEC-LT levels.(c) Avian immune response to pathogen (Task 4.4)In large-scale commercial trial in Drobex-Agro Sp. z o.o. (Poland), in ovo injection of prebiotics were found to have differential effects on levels of serum antibodies against 5 pathogens. While some prebiotics increased levels of some antibodies at the later time-points, while other were observed to increase antibody levels earlier and in certain cases, reduced levels at later time-points. There are a number of potential explanations for the results observed. For example, increases in antibody levels may indicate stable maintenance of antibodies against pathogen over time, while reductions in antibody may be indicative of a strong response early on, followed by a subsequent reduction or clearance of pathogen. Overall, these findings demonstrate that prebiotics has significant and differential effects in modulating peripheral immune response to pathogens. Moreover, these effects were observed in a large-scale production setting, thus validating the immunomodulatory effects of prebiotics in the commercial setting.(d) Intestinal morphology (Task 4.4)Differential changes in intestinal morphology and structure were observed for all prebiotics. Overall intestinal weight was significantly increased by DiNovo® and Bi2tos® (Day 42). Increases in surface area and width were achieved with DiNovo® and Lupin RFO on day 21. Thickness of muscularis and crypt depth were increased by Bi2tos® on day 21. The weight of intestines is associated with two factors, length of the intestine and wall thickness. Increased thickness of the intestine wall (including thickness of the muscularis) is positively related with the use of feed ingredients (better mixing of chyme and faster absorption of nutrients - increased motility of the intestinal villi), and therefore may provide a benefit to producers in this manner. 3.3.7 Improve meat quality (Task 3.3 4.3 4.4)A central aim of THRIVE-RITE was to develop prebiotic technologies which significantly improve meat quality according to key parameters relevant to producers, retailers and consumers alike. These targets were achieved for both pig & poultry, as outlined below.(a) Chicken meat quality• Ratio of polyunsaturated fatty acids (PUFA) to saturated fatty acids (SFA): P/S RatioIn ovo injection of DiNovo® in Task 4.3 was associated with an increase in the P/S ratio in breast muscle from 42 day old broilers. An increased P/S ratio is often recommended as a dietary means of preventing cardiovascular disease. • Meat colour: All prebiotics in Task 4.3 significantly improved meat colour by reducing redness at 45 min. post slaughter. DiNovo® & Lupin RFO reduced redness after 24 hours. • Pectoral Muscle size: All prebiotics influenced breast muscle size by increasing fiber diameter. Pectoral muscle weight was significantly increased by all prebiotics (Task 4.3). • Shelf-life: In the large-scale commercial trial (Task 4.4) Lupin RFO delayed progression of oxidation in meat up to day 6, while the untreated group showed significant signs of oxidation on Day 4. DiNovo® had positive effects on meat quality in the commercial trial, as it was associated with the lowest susceptibility to lipid peroxidation compared to the other prebiotic groups, and similar when compared to the control group. • Organoleptic properties: A trend towards increased intramuscular fat was achieved with DiNovo® (T4.3) an effect which may improve meat flavour, “juiciness” and tenderness.(b) Pork meat qualityThe aim of Task 3.4 was to develop prebiotics to increase the quality of pork meat, increase antioxidant content and shelf-life. These targets were successfully met. • Meat colour: NeoShield® provided significant improvements in pork quality to levels comparable with the commercial standard, Vitamin E, on day 3 of storage (reduced redness and reduced yellowness). NeoShield® and Bi2tos® significantly reduced yellowness compared to the control and to levels statistically indistinguishable from Vitamin E on day 3.• Meat shelf-life: Bi2tos® and NeoShield® significantly reduced levels of total viability counts (TVC) in LD muscle samples on day 5, to levels comparable with vitamin E. Bi2tos® and NeoShield® were also effective on day 10, while vitamin E in contrast, was ineffective at reducing TVC at this time-point. NeoShield® was the only treatment to reduce TVC on day 14, whereas vitamin E increased TVC at this time point. Lipid oxidation was significantly reduced by NeoShield® to levels comparable with Vitamin E on day 7, 11 and 14 of storage. While Bi2tos® was associated with increased lipid oxidation on day 11 and 14.• Antioxidant activity:All treatments significantly increased antioxidant activity in meat on day 4 of storage. NeoShield® was the only treatment to increase antioxidant activity at all time-points thereafter (day 7, 11, 14). Vitamin E on the other hand was ineffective beyond day 7. The only treatments to increase antioxidant activity at the final test date (day 14) were NeoShield® and Bi2tos®.3.3 Conclusions The stated aims of THRIVE-RITE were to develop products and application technologies which could achieve a number of clearly defined targets in pig and poultry meat production. From the conception of this project, the barometer of success was defined as the extent to which the problems identified could be solved. The table below summarises the significant results from THRIVE-RITE with respect to the main targets of the project, including:• Enhance productivity as measured by ADG & FCR -Target achieved• Reduce pig and poultry mortality - Target achieved• Reduce levels of pathogenic bacteria and viruses - Target achieved• Increase antibody transfer to offspring via maternal transfer - Target achieved• Improve antioxidant content in pig & poultry meat - Target achieved• Enhance gut health & peripheral immunity - Target achievedTHRIVE-RITE has validated a number of solutions to pressing problems facing food production in the EU. Knowledge as to the potential mode(s) of action of prebiotics has also been expanded considerably, thus providing significant contribution to knowledge generation. Key problems in EU agriculture and solutions identified by THRIVE-RITE include:• Rising food costs: NeoShield® improved feed conversion ratio (FCR) when used in cheaper, low energy diet. NeoShield® therefore, may provide producers with a means of switching to cheaper, more affordable diets.• Ban on antibiotics: Products were effective in reducing pathogens in a variety of situations. This was particular true in the case of challenge with Salmonella in finisher pigs and pathogens in poultry production such as Campylobacter and Enterobacteriaceae. In addition to providing a means of replacing use of antibiotics on countering bacterial pathogens, the data from THRIVE-RITE demonstrates their effectiveness against viral pathogens and parasites.• Reduced productivity: prebiotics were observed to increase performance in a range of conditions in pig and poultry production, in challenge, non-challenge and commercial scale production setting. Moreover, effects were observed via a number of application methods. The benefits of LactoShield® in pig production are particularly striking as it has been demonstrated that by feeding just one animal, the sow, positive performance effects are provided to ~12 progeny, thus providing substantial return on investment. Application of products via in ovo injection and via water proved highly effective in enhancing performance in poultry. • Ineffective alternatives: Products in THRIVE-RITE were generally robust compared to commercial positive controls such as zinc oxide, vaccines and Vitamin E. In this respect, THRIVE-RITE has identified products which can be used by producers who wish to move away from antibiotics usage or products which face increasing regulatory control in future (e.g. ZnO).In conclusion, these findings will provide the companies and food producers in Europe with novel means of enhancing growth, profitability and competitiveness. Potential Impact:Section 4: Potential impact, main dissemination activities and exploitation results 4.1 Potential impact 4.1.1 Background In the past, the requirement to increase productivity and agricultural output to feed the growing world population placed immense pressure on producers, resulting in a widespread culture of adding antibiotics to feed. While this was an effective means of improving animal health and output, it was banned in 2006 in the EU, due to concerns associated with overuse of antibiotics and potential effects on wider public health. However, the pathogens which affect pig and poultry farms have not gone away. These pathogens are highly evolved and are oftentimes highly species specific and successful in infecting their hosts. Without conventional treatments such as antibiotics, productivity has been significantly reduced. The EU’s failure to address this problem resulted in a dramatic rise in the use of therapeutic antibiotics, an issue of great concern to the European Medicines Agency (EMA; http://www.spiegel.de/international/europe/danish-pig-farmers-reduce-antibiotics-to-prevent-drug-resistance-a-933344.html). However, considerable foresight was demonstrated by the EU in 2012, by recognizing the potential of the THRIVE-RITE Consortium as having the capacity to generate effective alternatives to antibiotics. The THRIVE-RITE project is now completed and is expected to have immediate and long term impacts on the pig and poultry sectors, retailers and the consumer.THRIVE-RITE targeted the following problems that required viable and immediate solutions in the EU, both immediately and into the future: - Rising food costs- Lower producer margins due to lag effect- Ban on antibiotics- Reduced productivity- Limitations to current 'Solutions' to farming problem.Several products have emerged in THRIVE-RITE as having potential to solve these problems. The enterprises involved have established records in developing natural feed additives and this will ensure a prompt roll-out of the technologies to the end users. End users will benefit enormously from THRIVE-RITE, as the scientific information garnered will provide them with sufficient levels of detail in which to make informed decisions, a major requirement of large-scale producers in the industry. The robust experimental design and techniques employed by the leading scientific experts in the consortium, also reduces risk to the end users, as many previous unknowns with regard to product efficacy have been tackled at the commercial scale in this project. A summary of the findings pertinent to the project and the likely impacts, are described in the next section. With the successful roll-out of technologies from THRIVE-RITE, the project should serve as an excellent hallmark of how the EU should approach apparently insurmountable challenges and how to organize and implement research in a manner focused on the outcome- that is, solving the problem in a precise, viable and highly effective manner. 4.1.2 Impact SummaryTHRIVE-RITE has strong implications in terms of (a) sustainable means of pig and poultry production, (b) food safety, quality and shelf life and (b) human health and the environmental protection. THRIVE-RITE achieved the following major targets:• Enhance productivity as measured by ADG & FCR -Target achieved• Reduce pig and poultry mortality - Target achieved• Reduce levels of pathogenic bacteria and viruses - Target achieved• Increase antibody transfer to offspring via maternal transfer - Target achieved• Improve antioxidant content in pig & poultry meat - Target achieved• Enhance gut health & peripheral immunity - Target achieved(a) Sustainable means of pig and poultry production THRIVE-RITE has demonstrated that pig and poultry producers no longer need to depend on antibiotics in efforts to enhance the health of their flock or herd. Producers now have viable options available to them which have been compared with effective commercial standards such as zinc oxide. The products assessed in THRIVE-RITE have undergone a robust series of trials and a high level of scrutiny has been placed on efficacy and reproducibility, at levels expected and demanded from end users. The project has also assessed performance of the products when animals are under direct challenge by pathogens. The immune response to viral pathogen has also been assessed and efficacy of products has been compared with vaccine-based approaches to improving performance. This provides end users with proven means of managing herd health in a more cost effective way than relying exclusively on vaccine-based approaches. The advantage of THRIVE-RITE approach to product validation has been the requirement to conduct large-scale commercial trials, under academic supervision. This served to validate the scientific findings from the university to the ‘real-life’ scenario on the ground. Moreover, the stringent approach to experimental design has meant that significant findings at the commercial scale stand up to statistical scrutiny, a standard rarely achieved in the industry due to the inherent difficulties in allocating facilities, time, resources & the large number of pigs or chickens required for experimentation. This approach saved time and resources to validating the technologies and in this respect, ensures that findings have a more immediate impact. (b) Meat quality, shelf-life and safety One of the most striking findings from the THRIVE-RITE project is that the products assessed had significant effects in enhancing meat quality parameters, shelf-life and safety. In previous years, many have cited Vitamin E as the best product to enhance meat quality and shelf-life, largely due to its well-known antioxidant activity. Strikingly, NeoShield® conferred increases in antioxidant content in pork to levels similar to Vitamin E and over an extended time period. Antioxidant content was also increased with Bi2tos®. Moreover, both NeoShield® and Bi2tos® reduced total viability counts on pork muscle samples for a longer duration than Vitamin E, thus suggesting a role for these products in improving food safety. In fact, Vitamin E was observed to increase TVC after 14 days of storage, suggesting a negative effect of this vitamin on food safety. Lipid oxidation on pork meat was also reduced by NeoShield®, similar to Vitamin E, with positive effects on meat colour also observed with both NeoShield® and Bi2tos®. The impact of this research is that producers, retailers and end users alike can benefit from effects of these two new products which for the first time, have been shown to have positive effects on enhancing pork quality, antioxidant content and shelf life. THRIVE-RITE has also demonstrated that poultry meat quality is improved with the products. Interestingly, DiNovo®, Bi2tos® and Lupin RFO, were observed to differentially improve meat quality. All three improved meat colour and breast muscle fiber diameter. Lipid peroxidation was improved by Lupin RFO and DiNovo®. DiNovo® increased the ratio of polyunsaturated to saturated fatty acids in chicken meat. The advantage of the THRIVE-RITE approach to examining meat quality was the strong focus on the producer, retailer and end user. At present, there are very few viable means of improving meat quality. THRIVE-RITE has demonstrated that the prebiotics under study in this project provide novel means of improving meat quality, thus offering meat producers safer and cost effective means of increasing meat quality, shelf life and safety. (c) Human health and environmental protection• Human health:There are now major concerns of an imminent rise of drug-resistant microbes in animal production in the EU. This is due in part to the rapid increase in numbers of veterinarians prescribing antibiotics to farmers, a practice not only linked to the banning of in-feed antibiotics, but also associated with scandals surrounding the massive profits involved (Der Spiegel, http://www.spiegel.de/international/business/german-veterinarians-are-getting-rich-off-of-the-abuse-of-antibiotics-a-828289.html). The statistics show that huge levels of antibiotics are being administered by veterinarians in the EU, with Germany having been identified as being the worst offender in this regard (article published in Der Spiegel in 2013 http://www.spiegel.de/international/europe/danish-pig-farmers-reduce-antibiotics-to-prevent-drug-resistance-a-933344.html). The main reason for increased antibiotics in general is that producers do not have access to effective alternatives to in feed antibiotics following their ban in 2006. While Zinc oxide (ZnO) is currently an effective and widely used alternative, it faces increased scrutiny given the environmental implications. The majority of the 3Kg of ZnO added per tonne of pig feed is passed on through the faeces and into the environment. France banned ZnO in animal feed, more countries are expected to follow. THRIVE-RITE is expected to have significant impacts, as the products and technologies assessed have now been proven to effectively reduce pathogen load in pig and poultry. Producers will no longer have to rely on antibiotics or ZnO as means of increasing animal performance and profit. Viable alternatives have been identified. This is particular true in the case of challenge with Salmonella in finisher pigs and pathogens in poultry production such as Campylobacter and Enterobacteriaceae. In addition to providing a means of replacing antibiotics, the data from THRIVE-RITE demonstrates effectiveness of products against viral pathogens and parasites. This is significant, as producers will have access to products which not only counter bacterial pathogen, but also viral and parasitic infections which reduce performance and cause substantial economic losses. A striking finding from THRIVE-RITE was the effectiveness of the products in improving meat quality and this has potential for impacts on public health. DiNovo® was observed to increase P/S ratio in chicken meat, a proportion often recommended as a dietary means of preventing cardiovascular disease. These findings provide a novel means for developing methods of tackling public health issues and this may be particularly relevant to areas such as Northern Europe where the incidence of atherosclerosis and coronary heart disease are exceptional high. The efficacy of DiNovo® and Bi2tos® in improving antioxidant content and reducing TVC counts in meat may also provide a means of improving human health through benefits conferred by antioxidants themselves and by virtue of reducing pathogenic bacteria on the meat. • The environment:While Zinc oxide is cheap and highly effective way of increasing animal performance, it is not viable in the long terms due to concerns over environmental pollution. For the first time, animal producers will be offered a viable and effective means of moving away from zinc oxide by using effective and safe products such as those assessed in THRIVE-RITE.4.2 Impact of work packages (a) Work package 2:• Impacts on producers:- NeoShield: reduces Salmonella and increases productivity in finisher pigs (Task 2.3): Producers now have a product in NeoShield®, proven by Thrive-Rite to reduce Salmonella infection by almost 1 log and increase growth of finisher pigs. NeoShield® also modulated the immune system in the small intestine of treated pigs. There are few reliable products on the market which can achieve these effects. Thus, NeoShield® will provide producers with a new method of countering Salmonella and increasing productivity at the critical fattening stage of production. This is critical stage in the growth of finisher pigs, as increased performance translates into increased profit for the producer during this time.- DiNovo: increases broiler performance in flocks infected with Campylobacter (Task 2.4): Chicken producers will be able to increase average daily gain in broilers which are infected with Campylobacter, using a validated product, DiNovo®. There are few reliable natural products proven to increase broiler performance. Thus, producers will have access to a new, validated method of achieving these effects. - Immune modulation, gut health and beneficial bacteria (Task 2.3):Pig producers can access 2 products proven to modulate the immune system in pigs:- Both NeoShield® & Bi2tos® modulated the immune system in different ways. - Bi2tos® also increased beneficial bacteria, lactobacilli.There are few products, if any, on the market which can achieve these effects. • Impact on Society:Lower levels of Salmonella in the finisher pig achieved with NeoShield®, may reduce levels of this pathogen in pork meat, thereby providing a means of reducing the incidence of Salmonella poisoning in humans at the population-wide level. Retailers will benefit from enhanced reputation by having pork with reduced levels of Salmonella. Lower usage of antibiotics through use of NeoShield® can also offer a means of reducing the potential for drug-resistant pathogens to arise in animal production. This is an issue or growing importance in the EU given the extensive overuse of therapeutic antibiotics currently ongoing in countries such as Germany, an issue that has not yet been dealt with (http://www.spiegel.de/international/europe/danish-pig-farmers-reduce-antibiotics-to-prevent-drug-resistance-a-933344.html).(b) Work Package 3:• Impacts on producers:- LactoShield® enhances performance in weaned pigs:- Producers now have access to a product with an exceptional high level of validation to enhance the performance of the weaned pig. Effects can be observed in up to 12 offspring, by treating the sow alone. LactoShield® has been verified on a university and commercial scale to achieve the following (T3.3):- Enhance the life-time performance of pigs from weaning to slaughter age- Improved faecal consistency during the post-weaning period.- Reduce numbers of still-born pigs.- LactoShield® has been proven in the academic setting to improve performance and health in the face of challenge with ETEC or viral infection, as follows: - Improves faecal scores pre- and post-challenge with ETEC (T3.1).- Improve performance to levels equivalent to ZnO during the post-weaning period (T3.1).- Enhances the immune response in piglets infected with PCV-2 & PRRS virus (T3.2).- Increases ADG when compared to untreated animals and has a strong PCV-2 specific antibody response similar to that observed in the vaccine group (T3.2).- Increases serum IgA, IgG and IgM in first 60 days of piglet’s life (T3.2).Again, this gives producers access to a product which works by treating just one animal, the effects being observed in numerous offspring (up to 12 piglets). This is extremely novel and highly cost-effective for producers.- NeoShield® improves performance of weaned pigs when used in low energy diets:- In the UCD trial (D3.3) significant improvements in FCR were observed in pigs treated with NeoShield® and consuming a cheaper, low energy diet. Thus, NeoShield® provides producers with a means in which to prevent significant losses associated with more expensive diets.- Meat Quality:- T4.3 compared the efficacy of NeoShield® and Bi2tos® against Vitamin E in influencing meat quality, antioxidant content and TVC counts. These findings are significant, as producers now have access to 2 novel products, proven to enhance meat quality and related parameters. There are few, if any, safe and effective products on the market to achieve these effects. NeoShield® & Bi2tos® compete with Vitamin E to achieve the following when applied to finisher pigs:i. NeoShield:- Improves pork meat colour comparable to Vitamin E (D.3.4).- Reduced yellowness in pork meat to levels comparable to Vitamin E (D.3.4).- Reduction in TVC counts in pork LD muscle samples up to day 14 of storage, to levels greater than Vitamin E (D.3.4).- Reduced lipid oxidation in pork meat to levels similar to Vitamin E (D.3.4).- Increased antioxidant content, and to levels greater than Vitamin E (D.3.4).ii. Bi2tos:- Reduced yellowness in pork to levels comparable to Vitamin E (D.3.4).- Reduction in TVC counts in pork LD muscle samples up to day 10 (D.3.4).- Increased antioxidant content in pork meat (D.3.4).• Impact on Society:- Substituting antibiotics with LactoShield® may lower the potential for drug-resistant pathogens to arise in animal production, thus benefiting society.- Usage of zinc oxide may be reduced or replaced by using LactoShield®. Thus, the potential for environmental pollution associated with the use of ZnO is reduced.- Food safety is increased using NeoShield® or Bi2tos® in the finisher stage of pig production. This can provide consumers with meat with lower total viable counts & increased shelf-life.- Antioxidant content in meat from pigs treated with NeoShield® is higher than that achieved with vitamin E, thus providing potential health benefits to the consumer. - Retailers can improve their reputations as pork from NeoShield® or Bi2tos® treated pigs has lower TVC and increased antioxidant content.- Retailers and consumers alike can benefit from meat with longer shelf-life.(c) Work Package 4WP4 identified differential effects of prebiotics on enhancing health & performance, modulating the immune system, reducing pathogens and improving meat quality parameters in poultry. Each prebiotic proved beneficial but worked in different ways. This offers choice to producers when choosing a proven and effective prebiotics to enhance health and profitability. • Impacts on producers:- In ovo Technology and in water applicationsTHRIVE-RITE has successfully validated the efficacy of in ovo technology at both university and commercial scale in poultry production. Water applications of prebiotics have also been validated and provide benefits similar to, but slightly different to in ovo application. Thus, producers using traditional water delivery methods as well as producers using in ovo technology can both benefit from these findings. A critical impact of THRIVE-RITE is that a myriad of benefits are conferred by in ovo and water application of the prebiotics, including enhanced performance, modulation of the immune system, reduction in pathogen, increased beneficial bacterial, improved gastrointestinal health and improved meat quality.- DiNovo®, Bi2tos® and Lupin RFO: new products to reduce Clostridium:DiNovo®, Bi2tos® and Lupin RFO all reduced Clostridium in chickens when tested in a large scale commercial trial (D4.4). Producers affected by this pathogen have a choice of three new products to counteract this problem. This is likely to improve productivity as it may counteract the growth retardation effects of this pathogen.- Bi2tos: a new product to reduce Enterobacteriaceae:Bi2tos® was proven in the large-scale commercial trial to reduce levels of Enterobacteriaceae in chickens. This provides producers with a validated product for use in countering Enterobacteriaceae infection. There are few natural products, if any, proven to achieve these effects in commercial scale production of chickens.- DiNovo®, Bi2tos® and Lupin RFO: new products to reduce coccidial oocysts:DiNovo®, Bi2tos® and Lupin RFO all reduced levels of coccidial oocysts in chickens in a commercial trial. This gives producers access to novel products for countering infection with this common parasite. DiNovo® prevented high levels of infection early on and maintained oocysts at these levels, reducing them further at the last test date. This is significant, as global losses due to poultry coccidiosis has also been estimated as ~ $800M per annum (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3114971/ ), with >$3B is spent on controlling and preventing this disease annually, http://www.thepoultrysite.com/poultrynews/28036/high-cost-of-coccidiosis-in-broilers). Current methods of control include chemical, vaccination and ionophore, each with their limitations and problems, each with their advantages and disadvantages. For instance, while vaccination provides early resistance, birds become susceptible to necrotic enteritis infection. Chemical approaches may improve feed conversion early on; however, the parasite becomes resistant to chemical treatment. Ionophore provides resistance; however, oocyst populations build up, reducing feed conversion and growth rates, and can also be associated with gangrenous dermatitis (http://www.thepoultrysite.com/poultrynews/28036/high-cost-of-coccidiosis-in-broilers). Thrive-Rite has validated a new, prebiotic approach to counteracting Coccidiosis in broilers. This provides producers with new opportunities to counteracting this significant disease causing agent, and in an economically viable manner.- DiNovo: a new product to counteract Reovirus infection & mortality related losses:Reovirus infection has negative effects in chickens which manifest as viral arthritis/tenosynovitis, fibrinous tenosynovitis, stunting and atrophy of the pancreas. This results in general poor performance and reduced weight gain and thus, considerable economic losses. THRIVE-RITE has shown that DiNovo® increase chicken weight, reduces mortality and reduces viral load on the liver of chickens challenged with Reovirus (D4.5). This trial demonstrated the efficacy of DiNovo® in challenge situation and provides producers affected with Reovirus-related losses with a novel and effective means of countering this infection.- Modulating the immune system:Antibody responses specific to 5 pathogens were significantly modulated DiNovo®, Bi2tos® and Lupin RFO, including antibodies against:- Avian RhinoTracheitis (ART): causes ‘Swollen Head Syndrome’.- Infectious Bursal Disease (IBD): causes immunosuppression and economics losses due to secondary diseases and/or mortality thereafter.- Chicken anaemia virus (CAV): anaemia, immunosuppression & reduced growth.- Ornithobacterium rhinotracheale (ORT): respiratory disease, reduced weight gain.- Avian infectious bronchitis virus (IBV): reduces performance.Producers now have novel means of managing viral infection and health status of their flock through use of cost effective, scientifically and commercially validated prebiotic products.- Improving intestinal morphology:Intestinal morphology in poultry was differentially altered by DiNovo®, Bi2tos® and Lupin RFO. This provides producers with a choice of products which influence intestinal morphology and function in different ways. For example:- Bi2tos® increased intestinal weight (day 42) and increased thickness of intestine wall and crypt depth (D4.4). - Lupin RFO increased intestinal surface area and width (day 21; T4.4).- DiNovo® increased intestinal surface area, width (day 21) & weight (day 42; T4.4).End users concerned with intestinal health and looking to maximise nutrient absorption now have access to three new products which are proven to differentially influence gut weight and morphology. - Meat quality:Prebiotics successfully influenced poultry meat quality, shelf-life and related parameters in a university led trial (D4.3) and large-scale commercial trial (D4.4). - Meat colour: All prebiotics in Task 4.3 significantly improved meat colour by reducing redness at 45 min. post slaughter. DiNovo® & Lupin RFO reduced redness after 24 hours. This will provide producers with a competitive edge as they will be able to provide retailers with meat which retains a fresher colour for longer.- Pectoral Muscle size: All prebiotics influenced breast muscle size by increasing fiber diameter. Pectoral muscle weight was significantly increased by all prebiotics (Task 4.3). This is significant as producers will have a means of increasing yield or critical sections of chicken carcass, thus increasing profitability.- Shelf-life: Lupin RFO delayed progression of oxidation in meat up to day 6, while the untreated group showed significant signs of oxidation on Day 4 (D4.4). DiNovo® had lowest susceptibility to lipid peroxidation compared to the other prebiotic groups, and similar when compared to the control group. These findings are significant as they demonstrate that (a) the prebiotics do not negatively impact on meat quality- a key requirement of producers and (b) may be used to enhance shelf-life. - Ratio of polyunsaturated fatty acids (PUFA) : saturated fatty acids (SFA): In ovo injection of DiNovo® in Task 4.3 was associated with an increase in the P/S ratio in breast muscle from 42 day old broilers. Thus, producers will now have access to a product in DiNovo® which can be used to enhance this parameter. This will be very attractive to retailers as an increased P/S ratio is often recommended as a dietary means of preventing cardiovascular disease.- Organoleptic properties: A trend towards increased intramuscular fat was achieved with DiNovo® (T4.3) which may improve meat flavour, “juiciness” and tenderness. This may provide producers with a means of producing higher quality meat with improved taste, thus providing a competitive edge when selling to retailers.• Impact on Society:The major societal implication of WP4 is that poultry production can be undertaken in a more sustainable manner without the need for antibiotics or chemical treatments. This may reduce the potential for drug-resistant pathogens to arise in animal production, thus benefiting the broader society. In terms of food safety, Clostridium is the 3rd most common cause of food borne illness in the UK and USA. THRIVE-RITE has identified three products, DiNovo®, Bi2tos® and Lupin RFO, which can reduce this pathogen. This provides a means of reducing Clostridium without chemical or antibiotic treatments in poultry production. Bi2tos® also reduces levels of Enterobacteriaceae, a highly significant pathogen from a poultry & public human health perspective. Enhancements of meat quality may also have broader societal implications. A major finding from WP4 was that in ovo application of DiNovo®, proved effective in the face of Reovirus challenge in enhancing chicken growth and reducing mortality. This may have implications for public health in the future, as this trial proves in principal that DiNovo® is effective in the context of treating viral infection.(d) General Impacts The findings of THRIVE-RITE are extremely significant as they pave the way for a new approach to enhancing health in animals. We are entering an era of dramatic change in antibiotics use. Many experts consider that the practice of widespread antibiotics use is coming to an end, primarily out of necessity given the dramatic rise in multiple-resistant strains of pathogen and the fact that fewer and fewer effective replacements are available to some of the common antibiotics which were previously highly effective. (http://harvardmagazine.com/2014/05/superbug). THRIVE-RITE has validated a new approach to countering bacteria-based pathogens through use of prebiotic technology, modulation of the immune system and improving intestinal health. Moreover, viral pathogens continue to challenge animal and human health, with strains rapidly evolving and in many cases prove difficult to eradicate or manage through conventional vaccine technology. THRIVE-RITE has demonstrated that prebiotics can modulate the immune response to virus and even reduce mortality associated losses and performance issues in affected animals. As such, the findings in THRIVE-RITE open up a new scientific approach to dealing with viral pathogen. Another major impact of THRIVE-RITE is that the products can also be applied to enhance meat quality. There is currently a deficit in the provision of commercially viable means of achieving these effects in a manner safe to the consumer. There are also clear environmental benefits arising from THRIVE-RITE as producers now have access to products which will help them move away from using zinc oxide, a practice facing increasing regulation due to issues with heavy metal contamination. Another major implication to THRIVE-RITE is that novel maternal transfer mechanisms in which treatment of one animal (sow) can provide health benefits for up to 12 offspring, has been scientifically validated at the commercial level. This offers tremendous possibilities for improving animal health in a highly cost effective and efficient way. Likewise, the validated in ovo technology also have serious implications regarding an efficient means of administering prebiotics in poultry production. Critically, both maternal and in ovo technologies were validated at the university and commercial levels, a level of validation required by large-scale end users.Overall, THRIVE-RITE has achieved the aims of successfully validating the efficacy of the products under study in terms of their novel applications in animal production. Moreover, a considerable contribution to broadening scientific knowledge has also been made, both with regard to the hitherto unknown differential effects of different prebiotics and the novel application of this technology in terms of meat quality enhancement, maternal transfer to progeny, in ovo technology and managing viral pathogen.4.3 Dissemination activities 4.3.1 Dissemination Plan A key component of the dissemination plan was to ensure that producers, public and scientists alike were informed as to project targets. Targeting scientists and large-scale producers together was crucial, as a major barrier to solving the problems targeted by THRIVE-RITE are (a) a lack of communication between scientists and end users and (b) a lack of understanding between both groups as to requirements for validation at both university and commercial levels. These aims were explained to the public in a clear manner to ensure full understanding of how this research would benefit society in general. This achieved as follows:• Extensive local coverage via newspapers, magazines, radio and a number of documentaries. • Extensive international coverage with website articles discussing THRIVE-RITE in ≥11 countries and in at least 4 languages.• >60 dissemination activities in total.- >30 website articles- ~14 newspaper and magazine articles- >3 radio press releases and documentaries- >10 conferences and presentations. Several workshops and extensive teaching activities• Peer reviewed papers:- Manuscripts currently in preparation for publication in 2015 in high impact scientific journals.- 3 have been submitted so far (1 published, 1 in press, 1 under review as of 16/12/2014)4.3.2 Dissemination at Local levelThere was broad media coverage at local level via radio, newspapers, magazines and several documentaries. 4.3.3 Dissemination at International LevelVarious dissemination methods applied at international levels, including electronic media and scientific papers. List of Websites:For further information, please contact the THRIVE-RITE Project coordinator, as follows:John T. O’Sullivan, C.E.O. BioAtlantis Ltd., Kerry Technology Park, Tralee, Co. Kerry, Ireland. Tel. +353(0)66 711 8477e-mail: jtos@BioAtlantis.com , web: www.bioatlantis.com , Project website: www.thriverite.eu
