Progress 07/15/18 to 07/14/22
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Following is an outline of our major outreach plan: • Scientific peer-reviewed journal articles • Presentation at scientific meetings • Graduate student and postdoc training Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A graduate student was actively involved in all the studies. This graduate student received comprehensive training and obtained hands on experience on immunology, microbiology, and biochemistry. This project also partly supported a Postdoc to gain training in the areas of molecular microbiology and immunology. Finally, this project supported Dr. Jun Lin (PI) and his graduate student to attend national or international meeting for research presentation and professional development.. How have the results been disseminated to communities of interest?We have actively and effectively disseminated new information and procedures to scientists, producers, organizations, industries, government regulatory agency, and veterinarians. Following are some examples: Dr. Jun Lin (PI) gave multiple invited talks focused on the innovative technology developed and assessed in this project - the enterobactin-based immune intervention - to prevent and control colibacillosis and other pathogens in poultry. These talks were invited by Nanjing Agricultural University (July 17, 2018, China), APC Microbiome Institute/University College Cork (October 3, 2018, Ireland), The 3rd International Symposium on Alternatives to Antibiotics in Animal Production (Dec 16-18, 2019, Bangkok Thailand) and The First Symposium on Plant/Animal Health and Quality/Safety of Agro-Products (November 16-18, 2019, Ningbo, China) The lab personnel attended routinely attended annual Conference of Research Workers in Animal Disease (2018-2022) and American Society for Microbiology to give oral and poster presentations to disseminate new research findings to scientific communities. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
With support by this NIFA project, our collaborative team has made significant progresses toward developing novel Ent-based immune intervention to imrpove poultry health, which has led to 9 peer-reviewed high-impact journal articles (7 published and 2 under review), 11 abstracts presented in national conferences, and 2 thesis/dissertations. Following are brief summary of our accomplishments. Objective 1. Evaluate the immunogenicity of the Ent conjugate vaccine and its efficacy in protecting broilers from colibacillosis. We have completed a comprehensive characterization study focused Ent conjugate vaccine and obtained compelling evidence demonstrating that anti-Ent specific antibodies significantly inhibited APEC and other significant Gram-negative pathogens. Notably, we also performed a metabolomics study for the siderophore production profile (up to 14 different siderophores) of each of the 27 examined strains. These metabolomics data together with our work to sequence whole genome of the all examined strains further support our hypothesis that the developed Ent conjugate vaccine is an effective new vaccine against diverse APEC. Together, we obtained new and compelling microbiological evidence further demonstrating that the Ent-based immune intervention is a promising approach to control colibacillosis in poultry. We performed three large scale broiler trials to assess the efficacy of KLH-Ent conjugate vaccine to protect chickens from colibacillosis. The first two trials failed because the pathogenic APEC strains did not cause expected disease symptom in control groups, likely due to conditions changes related to the moving of Co-PDs to University of Georgia. After a series troubleshooting, we found the virulence plasmids in the APEC strains were lost, leading to attenuated virulence of the strains used for challenge. Subsequently, we have optimized APEC challenge model using new APEC strains with appropriate dose. In the third trial, we successfully evaluated the immunogenicity of the Ent conjugate vaccine and its efficacy in protecting broilers from colibacillosis. We observed that Ent conjugate vaccine elicited strong specific immune responses, leading to approximately 1,024 fold increase in the titer against the whole conjugate vaccine and 128 fold increase specifically directed against small Ent molecule when compared to unvaccinated groups. Upon APEC challenge, the O1 strain only caused sporadic lesions in lung, air sac, heart, liver, and spleen in control chicks, which were similar to those observed in vaccinated chicks. However, the highly virulent O78 strain caused extensive lesions in unvaccinated chicks; the KLH-Ent vaccination alleviated the lesions in different organs and significantly reduced the accumulative lesion scores. Together, the Ent conjugate vaccine could elicit strong specific immune responses in chickens and confer protection against APEC infection. To further increase the impact of the novel Ent conjugate vaccine on poultry health, we also evaluated immunogenicity of the Ent conjugate vaccine and its efficacy to control intestinal colonization of Campylobacter jejuni, a lead foodborne bacterial pathogen with chicken as a major reservoir. We observed that the Ent conjugate vaccine also could confer host protection against C. jejuni colonization, which provides a novel strategy for Campylobacter control in poultry and humans. Objective 2. Assess the Ent conjugate vaccine's ability to prevent colibacillosis in layers and yolk sac infections (YSI) in their hatchlings. Three chicken vaccination trials with different regimens have been performed to determine conditions for efficient production of anti-Ent egg yolk IgY in the past years. We have successfully optimized vaccination regimens for production of high-titer of Ent-specific egg yolk antibodies and demonstrated that the novel Ent conjugate vaccine could induce strong, specific, and robust immune response in chickens. With aid of the produced high quality egg yolk antibodies, we have performed several chicken trials for Objective 3. In addition, a large trial was performed to determine if vaccination of layers with the Ent conjugate vaccine would protect hatchlings against colibacillosis through vertical transfer of protective antibodies. Specifically, twelve breeder hens at 20 weeks of age were evenly assigned into 2 groups with one rooster (rode island red) being introduced to each group for fertilization. The layers in the treatment group were subcutaneously immunized with the Ent conjugate vaccine (100 μg per bird) while the other group served as non-immunization control. One week after the last vaccination, the eggs collected from each group were used for hatching. The newly hatched chicks (~30 birds per group) were housed separately. In each group, the chicks were intratracheally challenged with APEC O78 strain (107 CFU per bird) at 5 days of age. At two days post the challenge, all chicks were euthanized to assess lesions and APEC load in major organs. The egg yolks and blood samples (collected from both hens and hatchlings) were subjected to ELISA analysis. We observed that vaccination of the Ent conjugate vaccine significantly increased (16 fold) serum anti-Ent IgY titer when compared to the control group. The Ent-specific InY titers were in egg yolk and hatchlings treatment group were also significantly increased 32 fold and 8 fold, respectively, when compared to the control. However, the APEC challenge only led to very mild lesions in liver, lung, spleen, heart, and air sac in both groups with no significant difference observed with respect to both lesion scores and APEC load. In conclusion, vaccination of layers with the Ent conjugate vaccine could lead to successful vertical transfer of anti-Ent IgY from the vaccinated hen to her offspring. The APEC challenge model for young chicks needs to be optimized in the future. Objective 3. Determine the efficacy of passive immunization using Ent-specific egg yolk antibodies for protecting chickens from colibacillosis. With aid of the hyperimmune egg yolk powder obtained from Objective 2, we examined passive immunization of chickens with anti-Ent egg yolk powder for Campylobacter control. Interestingly, although we have demonstrated the Ent conjugate vaccine could significantly reduce C. jejuni colonization in chickens, passive immunization of chickens using Ent-specific egg yolk antibodies did not lead to reduced colonization of C. jejuni. We speculated that the orally administered egg yolk antibodies may be degraded in gastrointestinal tract in chickens. Therefore, we changed the focus of this objective by obtaining key information about functionality and in vivo bioavailability of orally administered egg yolk antibodies, a critical issue for the success of Ent-based passive immune intervention. We performed two chicken trials for both in vivo and ex vivo evaluation of stability of egg yolk IgY. Our findings indicated that the orally administered egg yolk antibodies were not stable in vivo. Specifically, egg yolk IgY could be substantially degraded in chicken gizzard, highly warranting development of effective approaches, such as encapsulation, for the controlled release and protection of orally administered egg yolk IgY in livestock. which was further demonstrated by ex vivo evidence.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Zeng, X., H.Wang, C. Huang, B. Gillespie, J. Lin. 2018. Efficacy of enterobactin conjugate vaccines to induce enterobactin specific egg yolk antibodies. Annual Conference of Research Workers in Animal Disease. December 1-4, Chicago
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Lin, J., X. Zeng, C.M.Logue, L.K. Nolan. 2018. Enterobactin-based immune intervention to control colibacillosis in poultry. Annual Conference of Research Workers in Animal Disease. December 1-4, Chicago
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Wang, H., J. Lin. 2019. A novel enterobactin conjugate vaccine induces high level of lipocalin-like antibodies for control of Gram-negative pathogens. American Society for Microbiology KY/TN Annual Conference. April 26-27, Nashville, TN.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2019
Citation:
Huiwen Wang, M.S. Thesis, University of Tennessee, Spring, 2019. Production and Characterization of Enterobactin specific Antibodies Induced by Novel Enterobactin Conjugate Vaccines.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2019
Citation:
Wang, H., X. Zeng, Y. Mo, B. He, H. Lin, J. Lin. 2019. Enterobactin-specific antibodies induced by a new enterobactin conjugate vaccine. Applied and Environmental Microbiology. 85:e00358-19.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lin, J., H. Wang, X. Zeng. 2019. Innovative enterobactin-specific egg yolk antibodies for controlling Gram-negative pathogens. The 3rd International Symposium on Alternatives to Antibiotics in Animal Production. Dec 16-18, Bangkok Thailand
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lin, J. 2019. Sustainability and safety of animal agriculture: emerging threats and innovative solutions. The First Symposium on Plant/Animal Health and Quality/Safety of Agro-Products. Ningbo, China, November 16-18, 2019.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Wang, H., and J. Lin. 2019. Enterobactin-specific antibodies: a novel tug-of-war weapon against Gram-negative pathogens. Annual Conference of Research Workers in Animal Disease. November 2-5, Chicago
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lin, J., X. Zeng, C.M.Logue, N. L. Barbieri, L.K. Nolan. 2019. Enterobactin-based immune intervention to control colibacillosis in poultry. Annual Conference of Research Workers in Animal Disease. November 2-5, Chicago
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Wang, H., X. Zeng, J. Lin. 2020. Enterobactin-specific antibodies inhibit in vitro growth of different Gram-negative bacterial pathogens. Vaccine. 38:7764-7773.
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Cui, Y., F.Guo, J. Guo, X. Cao, H. Wang, B. Yang, H. Zhou, X. Su, X. Zeng, J. Lin, and F. Xu. 2020. Immunization of chickens with the enterobactin conjugate vaccine reduces Campylobacter jejuni colonization in the intestine. Vaccines. 8, 747; DOI:10.3390/vaccines8040747
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Zeng, X., H. Wang, C. Huang, C.M. Logue, N. L. Barbieri, L.K. Nolan, J. Lin. 2021. Evaluation of the immunogenic response of a novel enterobactin conjugate vaccine in chickens for the production of enterobactin-specific egg yolk
antibodies. Frontiers in Immunology. https://doi.org/10.3389/fimmu.2021.629480
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Wang, H., X. Zeng, L. Cao, Q. He, J. Lin. 2021. Passive immunization of chickens with anti-enterobactin egg yolk powder for Campylobacter control. Vaccines, 9, 569. https://doi.org/10.3390/vaccines9060569
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Wang, H., X. Zeng, J. Lin. 2021. Ex vivo evaluation of enzymatic degradation of egg yolk IgY in chicken gastrointestinal tract. Frontiers in Immunology, 12:746831 https://doi.org/10.3389/fimmu.2021.746831
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
Cui, Y., F.Guo, J. Guo, X. Cao, H. Wang, B. Yang, H. Zhou, X. Su, X. Zeng, J. Lin, and F. Xu. 2020. Immunization of
chickens with the enterobactin conjugate vaccine reduces Campylobacter jejuni colonization in the intestine. Annual Conference of Research Workers in Animal Disease. December 5-8, 2020.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Wang, H., L. Cao, C.M. Logue, N. Barbieri, L.K. Nolan, J. Lin. 2021. Enterobactin-based immune intervention to control colibacillosis in poultry. Annual Conference of Research Workers in Animal Disease. December 3-7, 2021
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Wang, H., X. Zeng, F. Xu, L. Cao, C. M. Logue, N. L. Barbieri, L.K. Nolan, J. Lin. 2021. Enterobactin-based immune interventions against Gram-negative bacterial infections in chickens. Annual Conference of Research Workers in Animal Disease. December 3-7, 2021
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Cui, Y., H. Wang, F. Guo, X. Cao, X. Wang, X. Zeng, G. Cui, J. Lin, F. Xu. 2022. Monoclonal antibody-based indirect competitive ELISA for quantitative detection of Enterobacteriaceae siderophore enterobactin. Food Chemistry. 391:133241.doi: 10.1016/j.foodchem.2022.133241.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2022
Citation:
Wang, H., L. Cao, C.M. Logue, L.K. Nolan, J. Lin. 2022. Evaluation of immunogenicity and efficacy of the enterobactin conjugate vaccine in protecting chickens from colibacillosis. Vaccine.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2022
Citation:
Wang, H. , Q. Zhong, J. Lin. 2022. Hyperimmune egg yolk antibody for passive immunization: status, challenges, and prospects. Infection and Immunity.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Wang, H., C.M. Logue, L. K. Nolan, J. Lin. 2022. Assessment of an enterobactin conjugate vaccine in layers to protect offspring from colibacillosis. 102th Annual Conference of Research Workers in Animal Disease. January 20-24, 2023, Chicago.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2022
Citation:
Huiwen Wang, Ph.D., Summer, 2022. Enterobactin-based Immune Interventions against Gram-negative Bacterial Infections, The University of Tennessee
|
Progress 07/15/20 to 07/14/21
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Following is an outline of our major outreach plan: • Scientific peer-reviewed journal articles • Presentation at scientific meetings • Graduate student and postdoc training Changes/Problems:OBJECTIVE 3. We have performed two chicken trials to examine the stability and functionality of orally administered egg yolk antibodies. Given that solid evidence demonstarted the instability of egg yolk antibodies in the GI tract, it is not necessary to perform some proposed passivive immune protection of egg yolk antibodies against APEC infection in Objective 3. Our findings highly warrant future needs to develop practical approach to enhance in vivo stability of egg yolk antibodies (e.g. encapsulation) for passive immune protection against APEC infection. The new and significant findings from this objective research have led to following two peer-reviewed high impact journal articles: Wang, H., X. Zeng, J. Lin. 2021. Ex vivo evaluation of enzymatic degradation of egg yolk IgY in chicken gastrointestinal tract. Frontiers in Immunology, 12:746831 https://doi.org/10.3389/fimmu.2021.746831? Wang, H., X. Zeng, L. Cao, Q. He, J. Lin. 2021. Passive immunization of chickens with anti-enterobactin egg yolk powder for Campylobacter control. Vaccines, 9, 569. https://doi.org/10.3390/vaccines9060569 What opportunities for training and professional development has the project provided?A graduate student was actively involved in all the studies. This graduate student received comprehensive training and obtained hands on experience on immunology, microbiology, and biochemistry. This project also partly supported a Postdoc to gain training in the areas of molecular microbiology and immunology. How have the results been disseminated to communities of interest? Dr. Jun Lin (PI) and lab personnel attended Annual Conference of Research Workers in Animal Disease to give oral and poster presentations to disseminate new research findings to scientific communities. The results have led to 5 peer-reviewed research articles published on high impact journals (Frontiers in Immunology, Vaccine, and Vaccines) as well as 4 abstracts presented in annual Conference of Research Workers in Animal Disease. What do you plan to do during the next reporting period to accomplish the goals? We will repeat a chicken trial to confirm the observed protective efficacy of Ent-conjugate vaccine to control collisbacillosis in chickens (Objective 1) We will perform a large and complex trial to assess the Ent conjugate vaccine's ability to prevent colibacillosis in layers and yolk sac infections in their hatchlings as proposed in Objective 2. We will continue to generate peer-reviewed journal article and meeting abstracts resulting from this project.
Impacts
What was accomplished under these goals?
Objective 1. Evaluate the immunogenicity of the Ent conjugate vaccine and its efficacy in protecting broilers from colibacillosis. We have continued to examine significant potential of the Ent conjugate vaccine for broader applications to prevent and control diverse APEC and various Gram-negative infections. Specifically, we examined inhibitory effect of Ent-specific antibodies on the in vitro growth of three significant Gram-negative pathogens: Escherichia coli (n=27), Salmonella enterica (n=8), and Campylobacter spp. (n=6). Tthis study provides critical and compelling in vitro evidence supporting the feasibility of Ent-based immune interventions against several Gram-negative pathogens. The innovative findings have been published in a high-impact journal of Vaccine (Wang, H., X. Zeng, J. Lin. 2020. Enterobactin-specific antibodies inhibit in vitro growth of different Gram-negative bacterial pathogens. Vaccine. 38:7764-7773) Following two failed APEC trials reported in previous year, we have optimized APEC challenge model using new APEC strains with appropriate dose. Subsequently, we successfully performed a trial to evaluate the immunogenicity of the Ent conjugate vaccine and its efficacy in protecting broilers from colibacillosis. Specifically, specific-pathogen-free White Leghorn chicks were subjected to a crossed design with two variables, vaccination (with or without) and APEC challenge (APEC O1, O78, or PBS), leading to 6 groups (9 or 10 birds per group). Chickens were subcutaneously injected with PBS or KLH-Ent conjugate vaccine (100 ug per bird) at 7 days old, followed by booster immunization at 21 days old and subsequent intratracheal APEC challenge (108 CFU/bird of O1 strain, O78 strain, or PBS control) at 28 days old. At 5 days post-challenge, all chicks were subjected to necropsy to examine lesions and APEC load in major organs. Blood samples were collected at different time points and used for ELISA to monitor specific immune responses. Ileal and cecal samples were collected from each bird for microbiome analysis. We observed that Ent conjugate vaccine elicited strong specific immune responses, leading to approximately 1,024 fold increase in the titer against the whole conjugate vaccine and 128 fold increase specifically directed against small Ent molecule when compared to unvaccinated groups. Upon APEC challenge, the O1 strain only caused sporadic lesions in lung, air sac, heart, liver, and spleen in control chicks, which were similar to those observed in vaccinated chicks. However, the highly virulent O78 strain caused extensive lesions in unvaccinated chicks; the KLH-Ent vaccination alleviated the lesions in different organs and significantly reduced the accumulative lesion scores. Together, the Ent conjugate vaccine could elicit strong specific immune responses in chickens and confer protection against APEC infection. The findings from this trial has led to two abstracts. To further increase the impact of the novel Ent conjugate vaccine on poultry health, we also evaluated immunogenicity of the Ent conjugate vaccine and its efficacy to control intestinal colonization of Campylobacter jejuni, a lead foodborne bacterial pathogen with chicken as a major reservoir. The Ent-KLH conjugate vaccine could induce high-level of Ent-specific specific antibodies in chickens and confer host protection against C. jejuni colonization, which provides a novel strategy for Campylobacter control in poultry and humans. This study has led to one peer-reviewed article published in a high-impact journal (Cui et al. 2020. Immunization of chickens with the enterobactin conjugate vaccine reduces Campylobacter jejuni colonization in the intestine. Vaccines. 8, 747; DOI:10.3390/vaccines8040747) Objective 2. Assess the Ent conjugate vaccine's ability to prevent colibacillosis in layers and yolk sac infections (YSI) in their hatchlings. Three chicken vaccination trials with different regimens have been performed to determine conditions for efficient production of anti-Ent egg yolk IgY in the past years. We have successfully optimized vaccination regimens for production of high-titer of Ent-specific egg yolk antibodies and demonstrated that the novel Ent conjugate vaccine could induce strong, specific, and robust immune response in chickens. With aid of the produced high quality egg yolk antibodies, we have performed several chicken trials for Objective 3. In addition, the findings from this objective study has led to a high-impact journal articles (Zeng et al. 2021. Evaluation of the immunogenic response of a novel enterobactin conjugate vaccine in chickens for the production of enterobactin-specific egg yolk antibodies. Frontiers in Immunology. https://doi.org/10.3389/fimmu.2021.629480) Objective 3. Determine the efficacy of passive immunization using Ent-specific egg yolk antibodies for protecting chickens from colibacillosis. To obtain key information about functionality and in vivo bioavailability of orally administered egg yolk antibodies, a critical issue for the success of Ent-based passive immune intervention, we performed a chicken trial to examine passive immunization of chickens with anti-enterobactin egg yolk powder for Campylobacter control . Ourfindings indicated that the orally administered egg yolk antibodies were not stable in vivo, which was further demonstrated by ex vivo evidence. This study has been published recently (Wang et al. 2021. Passive immunization of chickens with anti-enterobactin egg yolk powder for Campylobacter control. Vaccines, 9, 569. https://doi.org/10.3390/vaccines9060569) To better understand the magnitude and dynamics of instability of egg yolk IgY in vivo, subsequently, we conducted comprehensive ex vivo analyses by spiking hyperimmune egg yolk IgY in fresh GI contents collected from five broilers at each sampling age (2, 4, or 6 weeks). We observed thategg yolk IgY could be substantially degraded in chicken gizzard, highly warranting development of effective approaches, such as encapsulation, for the controlled release and protection of orally administered egg yolk IgY in livestock. This new discovery was also published in a high-impact journal recently (Wang et al. 2021. Ex vivo evaluation of enzymatic degradation of egg yolk IgY in chicken gastrointestinal tract. Frontiers in Immunology, 12:746831 https://doi.org/10.3389/fimmu.2021.746831).
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Wang, H., X. Zeng, J. Lin. 2020. Enterobactin-specific antibodies inhibit in vitro growth of different Gram-negative bacterial pathogens. Vaccine. 38:7764-7773.
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Cui, Y., F.Guo, J. Guo, X. Cao, H. Wang, B. Yang, H. Zhou, X. Su, X. Zeng, J. Lin, and F. Xu. 2020. Immunization of chickens with the enterobactin conjugate vaccine reduces Campylobacter jejuni colonization in the intestine. Vaccines. 8, 747; DOI:10.3390/vaccines8040747
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Zeng, X., H. Wang, C. Huang, C.M. Logue, N. L. Barbieri, L.K. Nolan, J. Lin. 2021. Evaluation of the immunogenic response of a novel enterobactin conjugate vaccine in chickens for the production of enterobactin-specific egg yolk antibodies. Frontiers in Immunology. https://doi.org/10.3389/fimmu.2021.629480
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Wang, H., X. Zeng, L. Cao, Q. He, J. Lin. 2021. Passive immunization of chickens with anti-enterobactin egg yolk powder for Campylobacter control. Vaccines, 9, 569. https://doi.org/10.3390/vaccines9060569
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Wang, H., X. Zeng, J. Lin. 2021. Ex vivo evaluation of enzymatic degradation of egg yolk IgY in chicken gastrointestinal tract. Frontiers in Immunology, 12:746831 https://doi.org/10.3389/fimmu.2021.746831
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2020
Citation:
Cui, Y., F.Guo, J. Guo, X. Cao, H. Wang, B. Yang, H. Zhou, X. Su, X. Zeng, J. Lin, and F. Xu. 2020. Immunization of chickens with the enterobactin conjugate vaccine reduces Campylobacter jejuni colonization in the intestine. Annual Conference of Research Workers in Animal Disease. December 5-8, 2020.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Wang, H., L. Cao, C.M. Logue, N. Barbieri, L.K. Nolan, J. Lin. 2021. Enterobactin-based immune intervention to control colibacillosis in poultry. Annual Conference of Research Workers in Animal Disease. December 3-7, 2021
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Wang, H., X. Zeng, F. Xu, L. Cao, C. M. Logue, N. L. Barbieri, L.K. Nolan, J. Lin. 2021. Enterobactin-based immune interventions against Gram-negative bacterial infections in chickens. Annual Conference of Research Workers in Animal Disease. December 3-7, 2021
|
Progress 07/15/19 to 07/14/20
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Following is an outline of our major outreach plan: • Scientific peer-reviewed journal articles • Presentation at scientific meetings • Graduate student and postdoc training Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A graduate student was actively involved in all the studies. This graduate student received comprehensive training and obtained hands on experience on immunology, microbiology, and biochemistry. This project also partly supported a research scientist to gain training in the areas of molecular microbiology and immunology. Finally, this project supported Dr. Jun Lin (PI) and his graduate student to attend national or international meeting for research presentation and professional development. How have the results been disseminated to communities of interest?We have actively and effectively disseminated new information and procedures to scientists, producers, organizations, industries, government regulatory agency, and veterinarians. Following are some examples: Dr. Jun Lin (PI) gave multiple invited talks focused on the innovative technology developed and assessed in this project - the enterobactin-based immune intervention - to prevent and control colibacillosis and other pathogens in poultry. His talks were invited by The 3rd International Symposium on Alternatives to Antibiotics in Animal Production (Dec 16-18, 2019, Bangkok Thailand) and The First Symposium on Plant/Animal Health and Quality/Safety of Agro-Products (November 16-18, 2019, Ningbo, China) The lab personnel attended Annual Conference of Research Workers in Animal Disease (November 2-5, 2019, Chicago) to give oral presentations to disseminate new research findings to scientific communities. What do you plan to do during the next reporting period to accomplish the goals? We will perform chicken trials to evaluate the efficacy of the Ent conjugate vaccine in protecting chickens from colibacillosis. We will evaluate the efficacy of Ent-specific egg yolk powders to control collisbacillosis in chickens. We will publish two peer-reviewed journal article from this project and start to prepare another peer-reviewed journal article. We will submit abstracts and present findings in multiple national and international conferences.
Impacts
What was accomplished under these goals?
Objective 1. Evaluate the immunogenicity of the Ent conjugate vaccine and its efficacy in protecting broilers from colibacillosis. We have completed a comprehensive characterization study focused Ent conjugate vaccine and obtained compelling evidence demonstrating that anti-Ent specific antibodies significantly inhibited APEC and other significant Gram-negative pathogens. Notably, we also performed a metabolomics study for the siderophore production profile (up to 14 different siderophores) of each of the 27 examined strains. These metabolomics data together with our work to sequence whole genome of the all examined strains further support our hypothesis that the developed Ent conjugate vaccine is an effective new vaccine against diverse APEC. These findings has led to one research article submitted to the high impact journal Vaccine (resubmitted after modifications) and four abstracts presented in domestic and international conferences. We performed another broiler trial to assess the efficacy of KLH-Ent conjugate vaccine to protect chickens from colibacillosis. However, the experiment failed because the pathogenic APEC strains still did not cause expected disease symptom in control groups. Recently, the Co-PD (Dr. Cathy Logue at UGA) found the virulence plasmids in the strains were lost, leading to attenuated virulence of the strains used for challenge. New strains have been identified. Due to COVID-19 pandemic, new trial has been significantly delayed and will be performed in spring, 2021. Objective 2. Assess the Ent conjugate vaccine's ability to prevent colibacillosis in layers and yolk sac infections (YSI) in their hatchlings. We have completed the proposed studies to evaluate the immune response of layers upon vaccination with KLH-Ent conjugate vaccine and to prepare large quantities of Ent-specific egg yolk for the passive immunization work proposed in Objective 3. Specifically, we have immunized a total of 10 pullets with Ent-KLH conjugate vaccine using an optimized vaccination regimen. All eggs (post-immunization) have been collected and the egg yolks displayed significantly higher titer of Ent-specific IgY yolk (up to 2,048 fold) than the pre-immunization egg yolks. We have pooled all egg yolks and prepared large quantities of hyperimmune egg yolk powder that are used in Objective 3. The research work focused on production and characterization of egg yolk antibodies directed against enterobactin has led to one manuscript submitted to Frontiers in Immunology. Objective 3. Determine the efficacy of passive immunization using Ent-specific egg yolk antibodies for protecting chickens from colibacillosis. We have completed a preliminary chicken trial to determine in vivo availability of specific egg yolk powder supplemented in feed. This issue is critically important for interpretation of the findings from the proposed studies in this objective; however, to date, this issue has never been addressed in any of publications focused on application of egg yolk antibodies to prevent and control enteric animal pathogens. Briefly, 30 newly hatched chickens were assigned into two groups (15 birds per group) that receive starter feed or starter feed supplemented with 2% of egg yolk powder. At ages of 5d, 9d, and 14d, 5 birds were from each group for collecting blood and intestinal contents (from cecum and ileum, respectively). The IgY directed against Ent-KLH vaccine were assessed in all samples using ELISA. We observed that the specific IgY titer in the intestine and blood from treated chicks are very low, which is unexpected because we are confident the feed taken up by the chicks in treatment group has extremely high titer of specific IgY. Using different approaches, we will identify specific reasons causing poor in vivo availability of desired IgY.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lin, J., H. Wang, X. Zeng. 2019. Innovative enterobactin-specific egg yolk antibodies for controlling Gram-negative pathogens. The 3rd International Symposium on Alternatives to Antibiotics in Animal Production. Dec 16-18, Bangkok Thailand
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lin, J. 2019. Sustainability and safety of animal agriculture: emerging threats and innovative solutions. The First Symposium on Plant/Animal Health and Quality/Safety of Agro-Products. Ningbo, China, November 16-18, 2019.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Wang, H., and J. Lin. 2019. Enterobactin-specific antibodies: a novel tug-of-war weapon against Gram-negative pathogens. Annual Conference of Research Workers in Animal Disease. November 2-5, Chicago
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lin, J., X. Zeng, C.M.Logue, N. L. Barbieri, L.K. Nolan. 2019. Enterobactin-based immune intervention to control colibacillosis in poultry. Annual Conference of Research Workers in Animal Disease. November 2-5, Chicago
- Type:
Journal Articles
Status:
Under Review
Year Published:
2020
Citation:
Wang, H., X. Zeng, J. Lin. 2020. Enterobactin-specific antibodies inhibit in vitro growth of different Gram-negative bacterial pathogens. Vaccine (Resubmitted with modifications)
- Type:
Journal Articles
Status:
Submitted
Year Published:
2020
Citation:
Zeng, X., H. Wang, C. Huang, B. Gillespie, J. Lin. 2020. Production and characterization of enterobactin-specific egg yolk IgY for passive immunization. Frontiers in Immunology (Submitted)
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Progress 07/15/18 to 07/14/19
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Following is an outline of our major outreach plan: • Scientific peer-reviewed journal articles • Presentation at scientific meetings • Graduate student and postdoc training Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A graduate student was actively involved in all the studies. This graduate student received comprehensive training and obtained hands on experience on immunology, microbiology, and biochemistry. This project also partly supported a Postdoc to gain training in the areas of molecular microbiology and immunology. Finally, this project supported Dr. Jun Lin (PI) and his graduate student to attend national or international meeting for research presentation and professional development How have the results been disseminated to communities of interest?We have actively and effectively disseminated new information and procedures to scientists, producers, organizations, industries, government regulatory agency, and veterinarians. Following are some examples: Dr. Jun Lin (PI) gave multiple invited talks focused on the innovative technology developed and assessed in this project - the enterobactin-based immune intervention - to prevent and control colibacillosis and other pathogens in poultry. These talks were invited by Nanjing Agricultural University (July 17, 2018, China), and APC Microbiome Institute/University College Cork (October 3, 2018, Ireland). The lab personnel attended Annual Conference of Research Workers in Animal Disease (December 1-4, Chicago) and American Society for Microbiology KY/TN Annual Conference (April 26-27, Nashville, TN) to give oral and poster presentations to disseminate new research findings to scientific communities. What do you plan to do during the next reporting period to accomplish the goals? We will continue ongoing layer vaccination study to 1) determine immune response; and 2) produce large quantities of hyperimmune egg yolk antibodies. We will evaluate the efficacy of Ent-specific egg yolk powders to control collisbacillosis in chickens We will generate at least one peer-reviewed journal article from this project. We will submit abstracts and present findings in multiple national and international conferences.
Impacts
What was accomplished under these goals?
Objective 1. Evaluate the immunogenicity of the Ent conjugate vaccine and its efficacy in protecting broilers from colibacillosis. The quantitative in vitro growth assay was performed to examine the inhibitory effects of Ent-specific antibodies on the growth of diverse Avian Pathogenic E. coli (APEC) strains including those producing salmochelins; the purified lipocalin-2 was used as control. The Ent-specific antibodies significantly inhibited the growth of diverse E. coli strains under iron-restricted condition. Specifically, the Ent antiserum could lead to more than 7 log10 unit growth reduction, similar as that conferred by lipocalin-2. For the strains that also produce salmochelins, Ent-specific antibodies displayed significantly stronger inhibitory effects on the growth of E. coli than lipocalin-2. Together, we obtained new and compelling microbiological evidence further demonstrating that the Ent-based immune intervention is a promising approach to control colibacillosis in poultry. We also performed a broiler trial to assess the efficacy of KLH-Ent conjugate vaccine to protect chickens from colibacillosis; this vaccination trial is in a larger scale than the one proposed in proposal by including three heterologous strains of APEC isolates. However, likely due to the change of broiler breed for the challenge work, the three pathogenic APEC isolates did not cause expected disease symptom in control groups, leading to failure of this trial. We plan to perform dosemitric experiment to determine appropriate APEC dosage for challenge work in the future. Objective 2. Assess the Ent conjugate vaccine's ability to prevent colibacillosis in layers and yolk sac infections (YSI) in their hatchlings. We have initiated the proposed studies to evaluate the immune response of layers upon vaccination with KLH-Ent conjugate vaccine and to prepare large quantities of Ent-specific egg yolk for the passive immunization work proposed in Objective 3. Specifically, sufficient Ent conjugate vaccine was prepared. A total of 10 pullets were housed in the animal facility at UGA in July, 2019. All eggs (pre-immunization) were collected for preparation of control egg yolk antibodies. When all these layers become to actively lay eggs (expected in October, 2019), they will be immunized with KLH-Ent conjugate vaccine and produce sufficient hyperimmune egg yolk antibodies that will be used in Objective 3. Objective 3. Determine the efficacy of passive immunization using Ent-specific egg yolk antibodies for protecting chickens from colibacillosis. Not initiated yet.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Wang, H., X. Zeng, Y. Mo, J. Lin. 2018. Characterization of the enterobactin-specific antibodies induced by a novel enterobactin conjugate vaccine. Annual Conference of Research Workers in Animal Disease. December 1-4, Chicago
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Zeng, X., H.Wang, C. Huang, B. Gillespie, J. Lin. 2018. Efficacy of enterobactin conjugate vaccines to induce enterobactin specific egg yolk antibodies. Annual Conference of Research Workers in Animal Disease. December 1-4, Chicago
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Lin, J., X. Zeng, C.M.Logue, L.K. Nolan. 2018. Enterobactin-based immune intervention to control colibacillosis in poultry. Annual Conference of Research Workers in Animal Disease. December 1-4, Chicago
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Wang, H., J. Lin. 2019. A novel enterobactin conjugate vaccine induces high level of lipocalin-like antibodies for control of Gram-negative pathogens. American Society for Microbiology KY/TN Annual Conference. April 26-27, Nashville, TN.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2019
Citation:
Huiwen Wang, M.S. Thesis, University of Tennessee, Spring, 2019. Production and Characterization of Enterobactin-specific Antibodies Induced by Novel Enterobactin Conjugate Vaccines.
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