Periodic Reporting for period 1 - LIS (Evaluating effects of rumen originated lipopolysaccharide on the pathogenesis of subacute rumen acidosis)
Okres sprawozdawczy: 2022-06-01 do 2023-11-30
High-yielding dairy cows are often fed diets high in starch and low in fiber to boost energy intake and increase milk yield. However, such diets heighten the risk of subacute ruminal acidosis (SARA), a prevalent digestive disorder that negatively impacts animal health and herd profitability. SARA leads to reduced feed intake, decreased milk production, impaired rumen digestion, and severe health issues such as diarrhea, rumen mucosal damage, laminitis, inflammation, and liver abscesses. Studies have shown that high levels of fermentable starch in diets, such as corn silage and corn meal, are linked to increased SARA risk and reduced feed efficiency. Due to the high prevalence of SARA in Europe, which has resulted in substantial economic losses estimated at 994.4 million euros annually. Given the significant economic impact, understanding the causes of SARA and developing effective solutions to mitigate its occurrence is crucial.
Our project addresses this critical issue by focusing on the role of ruminal lipopolysaccharide (LPS) in SARA pathogenesis. Ruminal LPS, a major component of the outer membrane of gram-negative bacteria, is found in higher concentrations in SARA-affected cows due to bacterial lysis. Dosing LPS from E. coli into culture systems changes the bacterial community and increases certain bacteria. However, the potency of LPS from ruminal bacteria differs from that of E. coli, suggesting distinct impacts on the rumen environment. Meanwhile, the specific mechanisms underlying its involvement in SARA development remain unclear. By isolating and studying LPS from ruminal bacteria under SARA conditions, we aim to gain deeper insights into its mechanisms of action and its impact on rumen health.
The objectives of this project are to: 1. Isolate LPS from rumen bacteria by inducing SARA in a batch culture system;2. Compare the effects of ruminal bacterial LPS (LPS-R) with E. coli LPS (LPS-E) on ruminal fermentation, bacterial community composition, and function using 16S rRNA and RNA sequencing.
In conclusion, by elucidating these mechanisms, we can develop targeted interventions to mitigate SARA risk without compromising milk yield. This has the potential to not only improve animal welfare but also enhance the sustainability and profitability of dairy farming operations. Ultimately, our research has the power to transform dairy farming practices, reducing the prevalence of SARA and its associated economic losses while ensuring the continued supply of high-quality dairy products to meet global demand. Through collaboration with industry stakeholders, our findings can be translated into practical solutions that benefit both dairy farmers and society as a whole.
Our project addresses this critical issue by focusing on the role of ruminal lipopolysaccharide (LPS) in SARA pathogenesis. Ruminal LPS, a major component of the outer membrane of gram-negative bacteria, is found in higher concentrations in SARA-affected cows due to bacterial lysis. Dosing LPS from E. coli into culture systems changes the bacterial community and increases certain bacteria. However, the potency of LPS from ruminal bacteria differs from that of E. coli, suggesting distinct impacts on the rumen environment. Meanwhile, the specific mechanisms underlying its involvement in SARA development remain unclear. By isolating and studying LPS from ruminal bacteria under SARA conditions, we aim to gain deeper insights into its mechanisms of action and its impact on rumen health.
The objectives of this project are to: 1. Isolate LPS from rumen bacteria by inducing SARA in a batch culture system;2. Compare the effects of ruminal bacterial LPS (LPS-R) with E. coli LPS (LPS-E) on ruminal fermentation, bacterial community composition, and function using 16S rRNA and RNA sequencing.
In conclusion, by elucidating these mechanisms, we can develop targeted interventions to mitigate SARA risk without compromising milk yield. This has the potential to not only improve animal welfare but also enhance the sustainability and profitability of dairy farming operations. Ultimately, our research has the power to transform dairy farming practices, reducing the prevalence of SARA and its associated economic losses while ensuring the continued supply of high-quality dairy products to meet global demand. Through collaboration with industry stakeholders, our findings can be translated into practical solutions that benefit both dairy farmers and society as a whole.
Our project delved into the intricate realm of SARA, focusing on the pivotal role of ruminal LPS. Through a series of activities, the majority of the project's objectives were accomplished. We constructed an anaerobic gas station to facilitate controlled medium preparation and cultivation of rumen pure and mixed cultures. Collaborating with esteemed international partners, positioned our lab as a custodian of Italy's largest rumen culture collection, fostering groundbreaking research opportunities and international collaboration.
Transitioning to batch culture, we isolated ruminal LPS to unravel SARA's mysteries. We successfully induced SARA, validated by dynamic pH results showcasing sustained acidic condition. Detailed analysis unveiled altered fermentation profiles under SARA conditions, laying the groundwork for standardized SARA induction protocols in the batch culture, to improve experimental consistency and reliability. Furthermore, we determined the optimal peak concentration of free LPS for harvesting enriched free LPS media from rumen bacteria. Additionally, we standardized the processing procedure for collected enriched free LPS media from rumen bacteria.
Meanwhile, we meticulously compared the effects of ruminal bacterial LPS (LPS-R) with Escherichia coli LPS (LPS-E) on ruminal fermentation and bacterial community composition. Our findings clarify the role of LPS in SARA pathogenesis by revealing similar bacterial changes between LPS-R and LPS-E, resembling alterations observed in cows with SARA. Additionally, we observed that the effects of LPS on pathogenesis are time-dependent, with notable effects of LPS only observed after 24 hours of incubation. Importantly, LPS-E emerged as a practical alternative for future research, showcasing the robustness of our study results.
Moving forward, comprehensive analysis, including RNA sequencing, will deepen our understanding of LPS's intricate role in SARA. Collaboration with industry partners will be pivotal in developing innovative solutions for SARA control, particularly focusing on the development of LPS binding or absorbent products. These efforts will play a crucial role in safeguarding dairy cow health and ensuring the sustainability of the industry. The results of LIS were showcased at two major international conferences. Meanwhile, Dr. Dai has been invited to deliver talks at various prestigious research institutes and online live events. At least two scientific peer-reviewed papers from LIS will be published in open-access journals.
Transitioning to batch culture, we isolated ruminal LPS to unravel SARA's mysteries. We successfully induced SARA, validated by dynamic pH results showcasing sustained acidic condition. Detailed analysis unveiled altered fermentation profiles under SARA conditions, laying the groundwork for standardized SARA induction protocols in the batch culture, to improve experimental consistency and reliability. Furthermore, we determined the optimal peak concentration of free LPS for harvesting enriched free LPS media from rumen bacteria. Additionally, we standardized the processing procedure for collected enriched free LPS media from rumen bacteria.
Meanwhile, we meticulously compared the effects of ruminal bacterial LPS (LPS-R) with Escherichia coli LPS (LPS-E) on ruminal fermentation and bacterial community composition. Our findings clarify the role of LPS in SARA pathogenesis by revealing similar bacterial changes between LPS-R and LPS-E, resembling alterations observed in cows with SARA. Additionally, we observed that the effects of LPS on pathogenesis are time-dependent, with notable effects of LPS only observed after 24 hours of incubation. Importantly, LPS-E emerged as a practical alternative for future research, showcasing the robustness of our study results.
Moving forward, comprehensive analysis, including RNA sequencing, will deepen our understanding of LPS's intricate role in SARA. Collaboration with industry partners will be pivotal in developing innovative solutions for SARA control, particularly focusing on the development of LPS binding or absorbent products. These efforts will play a crucial role in safeguarding dairy cow health and ensuring the sustainability of the industry. The results of LIS were showcased at two major international conferences. Meanwhile, Dr. Dai has been invited to deliver talks at various prestigious research institutes and online live events. At least two scientific peer-reviewed papers from LIS will be published in open-access journals.
Our comprehensive investigation into the role of LPS in the pathogenesis of SARA is pioneering. We have not only delved into the effects of LPS on SARA pathogenesis but have also explored these effects irrespective of their origins. This groundbreaking approach suggests that LPS from E. coli can be utilized as a representative model for studying the impact of rumen bacterial LPS on SARA development. This departure from conventional methodologies expands the scope of research possibilities and enhances our understanding of SARA's underlying mechanisms.
Furthermore, our development of standardized SARA induction protocols in batch culture represents a significant advancement. These protocols ensure reproducible results and facilitate effective cross-study comparisons. By meticulously controlling various factors such as diet composition and rumen fluid sampling procedures, we can induce SARA consistently, thereby enhancing experimental reliability. This standardization addresses a crucial need in the field and sets a new standard for experimental methodologies in rumen health research.
Additionally, the project holds significant socio-economic implications and broader societal benefits. By uncovering the role of LPS in SARA, we're not just transforming dairy farming practices but also enhancing animal welfare and health outcomes. The ability to regulate LPS levels stands to greatly enhance farm profitability while simultaneously mitigating environmental impacts by specific products, such as methane emissions. Moreover, our research findings play a crucial role in informing policy-making processes, providing policymakers with invaluable insights into the management of SARA and its associated risks. This, in turn, fosters collaboration within the industry, leading to the development of effective strategies and interventions. These collaborative efforts translate into tangible benefits for stakeholders across the agricultural sector, driving advancements in farming practices and promoting sustainability.
Furthermore, our development of standardized SARA induction protocols in batch culture represents a significant advancement. These protocols ensure reproducible results and facilitate effective cross-study comparisons. By meticulously controlling various factors such as diet composition and rumen fluid sampling procedures, we can induce SARA consistently, thereby enhancing experimental reliability. This standardization addresses a crucial need in the field and sets a new standard for experimental methodologies in rumen health research.
Additionally, the project holds significant socio-economic implications and broader societal benefits. By uncovering the role of LPS in SARA, we're not just transforming dairy farming practices but also enhancing animal welfare and health outcomes. The ability to regulate LPS levels stands to greatly enhance farm profitability while simultaneously mitigating environmental impacts by specific products, such as methane emissions. Moreover, our research findings play a crucial role in informing policy-making processes, providing policymakers with invaluable insights into the management of SARA and its associated risks. This, in turn, fosters collaboration within the industry, leading to the development of effective strategies and interventions. These collaborative efforts translate into tangible benefits for stakeholders across the agricultural sector, driving advancements in farming practices and promoting sustainability.