Progress 10/01/08 to 09/30/13
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Characterization of a direct fed microbial model for the study of allocthonous bacteria on the innate and systemic immune system.
Publications
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Progress 10/01/09 to 09/30/10
Outputs
OUTPUTS: Studies have continued on methods of manipulating commensal bacteria within the gastrointestinal tract in such a way as to increase animal health and productivity. Emphasis has been placed on the changes in whole broiler metabolism and energy expenditures caused by alterations in gastrointestinal flora. This has necessitated the development of new techniques that allow for the estimation of energy expenditures by individual cells within the immune system. To date, this research program has shown that the introduction of direct fed microbials into the gastrointestinal tract will alter energy partitioning in the whole animal. This is especially true of the immune system and the intestinal tract. The data suggests that certain DFMs decrease immune energy expenditures in birds not undergoing an immunological challenge but does cause a dramatic shift in immune system energy expenditures in immune challenged birds. This is also accompanied by increased expression of certain cytokines and antibodies. One interesting detail noted in the present work is that the amount of DFM actually being fed represents a very small number of bacterial cells and DNA in comparison to the resident population of bacterial within the gastrointestinal system. This has led our laboratory to speculate on specific mechanisms that may redefine our thinking of the role of DFMs and probiotics in relation to host animal tissues. These findings have been communicated to the scientific community in invited talks both at national and international symposia as well as in abstract form at scientific meetings PARTICIPANTS: Warren J. Croom, Jr., Ph.D. Co-Principal investigator, Professor of Poultry Science Matthew D. Koci, Ph.D. Co-Principal investigator, Assistant Professor of Poultry Science TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
1. This program will allow increasingly better assessment of the effects of changes in bacterial function in the gastrointestinal tract and changes in metabolism and energy expenditures in the host animal. 2. Development of quicker ways of assessing DFM and probiotic effects in animal's will enhance the velocity of our development of new DFM and probiotic consortia that will either prevent or treat diseased states in the gastrointestinal tract and other tissues. 3. As we elucidate mechanisms of how small numbers of DFM or probiotic bacteria influence host animal metabolism and immune function, we will be able to develop highly specific treatment modalities that could be more efficient than the current use of DFM or probiotic live bacterial cells. This approach could be used to help eliminate or reduce the need for antibiotic use in food animal production systems.
Publications
- Qui R.Q., Croom J., Koci M.D., 2010. Nutrition, microbiota, and immunity. Pancosma Worldwide Scientific Exchange Proceedings. Paris, France
- Qui, R.Q., Croom, J., Koci M.D., 2010. Promoting animal and human health through direct manipulation of the intestinal flora. Minnesota Nutrition Conference Proceedings. Owatonna, Minnesota
- Qui R.Q., Croom J., Koci M.D., 2010. Direct fed microbial supplementation alters host's immune response and repartitions energy to the immune system. ADSA-PSA-AMPA-CSAS-WSASAS-ASAS Joint Annual Meeting. Denver, Colorado
- Qui R.Q., Croom J., Koci, M.D., 2010. Direct fed microbials influence the development and ultrastucture of the pre- and post-gastric intestinal tract. Microbial Communities in Health, Disease and Our Environment.(Abstract) Chapel-Hill, North Carolina
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Progress 10/01/08 to 09/30/09
Outputs
OUTPUTS: Work has continued to characterize the impact of the probiotic consortium, Primalac, on whole broiler and broiler organ system energetics. Efforts in our laboratory have confirmed our previous observations that that this probiotic does not affect whole-body energy consumption but does affect energy expenditures and repartitioning portions of the intestinal tract and the systemic immune system. Our work has shown that resting immune cells in probiotic fed animals have higher levels of ATP whilst non-specifically challenged animals have lower intracellular ATP. This, along with higher circulating levels of antibodies, suggests that probiotic fed broilers mount a more vigorous immune response to non-self antigens. Our research continues to work to identify the mechanism of action of the probiotic organisms on the host-bird tissues. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Research with this probiotic consortium model, Primalac, holds excellent promise for the elucidation of the mechanisms through which probiotics, introduced into the intestinal tract, evoke positive changes in immune function in the broiler. This information could lead to the development of new, non-chemotherapy, methods of preventing and treating disease in broiler and other poultry flocks.
Publications
- J. Croom, M. Chichlowski, M. Froetschel, B. W. McBride, R. Qui and M.D. Koci 2009 The Effects of Direct Fed Microbial Primalac, or Salinomycin Supplementation on Intestinal Lactate Isomers and Cecal Volatile Fatty Acid Concentrations in Broilers. International J. Poultry Science. 8:128-132.
- Steele, M. A., O. AlZahal, S. E. Hook, J. Croom and B.W. McBride 2009 Ruminal acidosis and the rapid onset of ruminal parakeratosis in a mature dairy cow: a case report. Acta Veterinaria Scandinavica. 51: 39-45.
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Progress 10/01/02 to 09/30/08
Outputs
OUTPUTS: During the life of this project we investigated the effects of peptide YY (PYY) and probiotics in maintaining the health and well-being of broilers and turkey poults. Peptide YY was investigated as a means of enhancing maturation of the poult or chick intestinal tract, hence increasing bird robustness and ability to resist enteric infections. In ovo administration of peptide YY was shown to increase glucose transport in hatchling turkey poults up through one week of age. Subsequent injection with PYY at the dosage of 150ug/kg body weight would sustain this increase in intestinal absorption. Additionally we documented a genotypic influence on the effects of in ovo administration of PYY on subsequent changes in the rate of jejunal glucose uptake. Turkey poults from lines selected for growth required a larger dosage of PYY to show increases in intestinal absorption than turkey lines selected for egg production. Further research is planned to delineate PYY's mode of action in the turkey and broiler. We selected a commercial probiotic or direct fed microbial as a model to describe the mode of action of autochthonous bacteria in the gastrointestinal tract as related to enhanced intestinal and whole-body health as well as growth and performance. We systematically estimated the effects of this probiotic consortium in broilers on the manner of colonization within the gastrointestinal tract as view with scanning electron microscopy, whole-bird energy expenditures using closed circuit indirect calorimetry, individual tissue oxygen consumption, ileal glucose and amino acid absorption, production of select cytokines in the gastrointestinal tract and the effects of probiotics on whole body immune function and energy partitioning into various tissues associated with growth and immune function. Our studies demonstrated that feeding this probiotic consortium resulted in colonization of lactobacilli that existed in a very close spatial relationship with intestinal enterocytes. Furthermore, supplementation of this consortium increased intestinal cytokine production, systemic cell mediated and humoral immune response. Interestingly, the body of control broilers fed the consortium partitioned less energy into the immune system. In contrast, broilers fed the consortium, given a non-specific challenge, repartitioned more energy into immune tissues, increased both cell-mediated and humoral immune responses, but did not result in the birds expending more whole-body energy. Further studies are planned with this consortium using specific pathogen challenges. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
This project has demonstrated that the use of naturally occurring peptides continue to be a putative manner in which to improve the health and performance of broilers and poults. More mechanistic information is needed to reduce this information to practice within the industry. Of paramount importance is the need to develop methods of extending the benefits of the initial administration of such intestinal modulators as peptide YY. Genotype and gender variation need to be further described. Our investigations into the mechanisms action of probiotics have conclusively demonstrated that some consortiums evoke potentially beneficial immune responses. As the mechanisms of actions on the immune system become better understood, there is no doubt that effective prophylactic and therapeutic probiotic consortia will be able to be developed for use in poultry production systems to reduce disease and increase bird performance.
Publications
- Chichlowski,M. J. Croom, R. Qui, B.W. McBride and M.D. Koci. Direct Fed Microbial, Primalac, Supplementation and Jejunal Glucose and Proline Transport in Broiler Chickens 2008 International Journal of Poultry Science 7: 1163-1166
- Fairchild, A.S., J. Croom, J.L. Grimes and W.M. Hagler, Jr.Effect of ASTRA-BEN 20 on Broiler Chicks Exposed to Aflatoxin B1 or T2 Toxin 2008 International Journal of Poultry Science 7: 1147-1151
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Progress 10/01/06 to 09/30/07
Outputs
OUTPUTS: Activities: Several experiments were conducted to further elucidate the impact of supplementation of the direct fed microbial model (DFM), Primalac, on the energetics of immune response in broiler chickens. Broiler chicks were fed the DFM from hatch until 26 days of age. During that period, the effects of DFM supplementation on whole body energy production as well as the energy status of white blood cells and nitrous oxide production by macrophages were estimated in a setting of a non specific (sheep red blood cell) antigen challenge. Other experiments were conducted on the effects of this DFM supplement on the oxidative stress in the membranes and mitochondria of the ileum and cecum of broiler chicks fed the supplement for 21 days. A final experiment examined the effects of an experimental enzyme feed supplement, keratinase, on energy expenditures and intestinal absorptive cell histology and ultrastructure in broiler chicks. The results of all experiments were presented at
poster sessions during the annual meetings of the Poultry Science Association during July at San Antonio, Texas. All experiments were conducted as a portion of the doctoral training of two students.
PARTICIPANTS: Croom, J., Department of Poultry Science, North Carolina State University (PI) Daniel, L. Department of Poultry Science, North Carolina State University, (collaborator) Koci, M.D., Department of Poultry Science, North Carolina State University (PI) Qui, R. Department of Poultry Science, North Carolina State University, (Ph.D. student trainee)
TARGET AUDIENCES: Academics in poultry and animal science, nutrition and immunology: Poultry Feed Industry Technical Support (data used in industry presentations to explain mode of action of direct fed microbials or probiotics to producers within the USA as well as other countries)
PROJECT MODIFICATIONS: None
Impacts
Change in Knowledge: These experiments were the first to demonstrate that although the feeding of direct fed microbials (DFM) had no impact on whole body energy consumption, DFM supplementation did affect the partitioning of energy amongst and within organ and tissue systems within the body of broiler chickens. Direct fed microbial supplementation decreased resting, antigen unchallenged energy states in white blood cells whilst increasing energy states and antibody output during challenges with sheep red blood cells. Similarly, thymus gland energy expenditures of DFM supplemented broilers were not different during control states (non sheep red blood cell challenge) from non-DFM supplemented broilers. After challenge with sheep red blood cells (challenged state), the thymus gland of DFM supplemented broilers expended greater amounts of energy compared to broilers that had no supplementation. DFM supplementation resulted in no change in the oxidative stress of the
membrane and mitochondria of the ileum, although DFM did increase oxidative stress in the membranes of the cecum of young broilers. Supplementation with the experimental enzyme, keratinase, had no effects on broiler chick ileal histology, although changes were noted in the ultrastructural surface of enterocytes of birds supplemented with this enzyme.
Publications
- Chichlowski. M., W. J. Croom, F. W. Edens, B. W. McBride, R. Qiu, C. C. Chiang, L. R. Daniel, G. B. Havenstein, and M. D. Koci. 2007. Microarchitecture and Spatial Relationship Between Bacteria and Ileal, Cecal, and Colonic Epithelium in Chicks Fed a Direct-Fed Microbial, PrimaLac, and Salinomycin. Poultry Science. 86: 1121-1132.
- Chichlowski. M., J. Croom, B. W. McBride, L. Daniel, G. Davis, and M. D. Koci Direct-Fed Microbial PrimaLac and Salinomycin Modulate Whole-Body and Intestinal Oxygen Consumption and Intestinal Mucosal Cytokine Production in the Broiler Chick. 2007. Poultry Science 86: 1100-1106.
- Chichlowski, M., J. Croom, B.W. McBride, G.B. Havenstein and M.D. Koci Metabolic and Physiological Impact of Probiotics or Direct-Fed-Microbials on Poultry: A Brief Review of Current Knowledge. 2007. International Journal of Poultry Science 6 (10): 694-704.
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Progress 10/01/05 to 09/30/06
Outputs
Although many studies show that supplementation of probiotics reducing the incidence of enteric disease and enhance growth and performance, the mechanisms of action of probiotics in poultry are poorly understood. If the mechanisms of action can be understood, more efficacious probiotic consortium can be designed to enhance poultry health and decrease the incidence of disease. A series of studies was conducted to investigate the effects of a mixed consortium of direct fed microbials or probiotics on broiler metabolism and immune status. In the first study, the effects of a probiotic consortium was compared to a control diet and one substituted with salinomycin, a commonly used antibacterial and anti-protozoal used by the poultry industry. Probiotic supplementation significantly decreased whole-body energy expenditures as well as ileal oxygen expenditures compared to salinomycin supplementation. Numerical trends were observed for decreased whole-body and ileal oxygen
consumption as compared to controls. Probiotic supplementation resulted in decreased production of intestinal mucosal cytokines as compared to control and salinomycin supplemented diets. Probiotics also increased passive glucose and proline transport in the jejunum and ileum. Histomorphometric studies demonstrated that probiotic supplementation increased the number of intestinal goblet cells, jejunal villi surface area and muscle thickness. Scanning electron microscope studies demonstrated that probiotic supplementation resulted in increased attachment of Lactobaccillus-like organisms to the surface of the jejunum, ileum and cecum. Additionally, close spatial relationships were noted between these organisms and the goblet cells and crypts. Probiotics also seemed to alter the amount and consistency of intestinal mucus. Preliminary studies noted that probiotic supplementation had no effect on the amount of free-radicals associated with enterocyte mitochondria. Other pilot studies
suggest that probiotic supplementation increased anti-body production after specific and non-specific antigen challenges to the broilers without increasing whole-body energy expenditures. Antigen challenges increased demand for ATP within the white blood cells. These data suggest that our model probiotic consortium may enhance intestinal and whole-body immunity without altering whole-body energy consumption. This might be accomplished by an unknown repartitioning of available energy within the broiler. Further studies are planned to elucidate the action of probiotics on immune function and the energetic expenditures of portions of the immune system.
Impacts
Our studies have demonstrated that supplementing the diets of broilers with probiotics or friendly bacteria can stimulate both whole bird and intestinal immune response. Additionally, supplementation of probiotics to poultry may increase the effectiveness to routine disease immunization and their resistance to other diseases for which their are no vaccines or treatments. Studies are currently underway to better understand how probiotic supplementation works within the bird. Achieving this would allow the design of more effective probiotics and healthier poultry flocks. This is an issue of emerging importance since pressure from the scientific community and general public will result in a ban of antibiotics from use as routine poultry disease preventatives. Additionally, increasing awareness by the public of possible links between poultry diseases and those that can be transmitted to humans is sparking increased public concern over the health of the nation's poultry
flocks. Present and future generations of probiotics will likely play a vital role in maintaining the health of the nations poultry flocks and limit their impact on the occurrence of human disease.
Publications
- Croom W.J., Jr.,Decubellis, J.,Coles B.A., Daniel L.R.,Christensen V.L. 2006. Effect of in ovo peptide YY on the ontogeny of glucose transport in turkey poults. Int. J. Poul. Sci. 5:128-133
- Fan Y.K.,Croom W.J.,Daniel L.,McBride B.W.,Koci M.,Havenstein G., Eisen E.J. 2006. Effect of genotype on whole-body and intestinal metabolic response to monensin in mice. Asian-Aust. J. Anim. Sci.19:554-560
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Progress 10/01/04 to 09/30/05
Outputs
Currently, there is no cohesive, integrated understanding within the scientific community, of how allocthonous bacteria, such as are found in probiotics (Pro) or direct-fed-microbials (DFMs) contribute to general and gastrointestinal health of poultry. An understanding of this phenomenon is essential to the future profitability of the poultry industry since it is generally accepted that Pro and DFMs can afford some of the same protection from enteric and systemic disease as the prophylactic antibiotics that are currently being phased from use. Broiler chicks, 21 days of age, were used to investigate the mechanisms of action of direct-fed-microbials (DFMs) on gastrointestinal health, whole body metabolism and growth and feed efficiency. We have found that a DFM, consisting primarily of lactobacilli, had no effect on dry matter intake, decreased whole body energy expenditures by 17% and intestinal energy expenditures by 47% as compared to control birds or birds fed
salinomycin. Concomitant to these decreases in whole-body and gastrointestinal energy expenditures, production of both pro and anti-inflammatory cytokines were reduced. Analysis of gastrointestinal fermentation products, although not fully completed, have shown a 50% decrease in L-lactate acid concentration of the jejunum of DFM birds compared to those of control and salinomycin treated birds. Scanning electron microscopy (SEM) of the ileums of all treatments demonstrated significant colonization by lactobacilli of the villi crypts surface areas near goblet cells in the DFM treated birds. This shift in bird energetics associated with the use of the DFM, at both the whole-body and tissue level, is dramatic and speaks to a complex interaction between the organisms contained in the DFM and the host bird tissues. SEM studies support this conclusion. Further studies are underway to more fully examine the mechanisms associated with these beneficial metabolic and physiological responses.
Impacts
We have been the first to demonstrate, that the use of direct-fed-microbials or probiotics, decrease whole-bird energy utilization as well as intestinal energy utilization. This discovery has the potential to exert enormous impact on the profitability of the North Carolina poultry industry, especially broilers, in two ways. First, the decreased whole-bird energy utilization suggests that direct-fed-microbials have the potential to increase the energetic efficiency of feed utilization which will result in lower fed costs. Second, the discovery that direct-fed-microbials decreased intestinal energy expenditures while simultaneously down- regulating intestinal, mucosal, cytokine production, strongly supports the protective effects direct-fed-microbials have on the health of the gastrointestinal tract, thus preventing death from intestinal diseases.
Publications
- Belstra, B.A., Flowers, W. L., Croom, W. J., Degroot, J. and See, M. T. 2005. Urinary excretion of collagen degradation markers by sows during postpartum uterine involution. Anim. Reprod. Sci. 85:131-145.
- Fairchild, A.S., Grimes, J. L., Porter, J.K., Croom, W.J., Daniel, L.R., Muller, M. and Hagler, W.M., Jr. 2005. Effects of diacetoxyscirpenol and fusaric acid on poults: Individual and combined effects of dietary diacetoxyscirpenol and fusaric acid on turkey poult performance. Int. J. Poult. Sci. 4:350-355.
- Survana, S., Christensen, V., Ort, D.T. and Croom, W.J., Jr. 2005. High levels of dietary carbohydrate increase glucose transport in poult intestine. Comp. Biochem Physiol A Mol Integr Physiol. 141: 257-263.
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Progress 10/01/03 to 09/30/04
Outputs
We have initiated a multi-disciplinary program with the goal of collecting data that will allow for the construction of a rationale paradigm for the design of probiotics and direct-fed microbials in poultry and other domestic livestock species. We emphasize measuring the effects of probiotic on nutritional physiological parameters such as whole body energetics, intestinal energetics, intestinal nutrient absorption, intestinal nutrient transporter density, intestinal histomorphometrics, gastrointestinal volatile fatty acid and lactate production and mucosal and whole-body immunologic function. Special care is being given to ensuring the patency of controls and the proper colonization of treatments using RNA profiles unique to the consortia being studied. Additionally scanning electron microscopy is being used to study possible unique associations between intestinal structures and adherent strains of allocthonous bacteria. Initial studies have been completed and data
are currently being summarized and interpreted. Future studies will explore the possible effects of probiotics on ion exchange across the lower intestinal tract, cecum and colon as well as the production of certain long-chain fatty acids that may have a direct stimulatory effect on the avian immune system.
Impacts
Probiotic and direct-fed microbial development have largely been based on trial and error formulations which have been screened using performance-based growth trials. Construction of a list of the actions of gastrointestinal bacteria that exert a desirable host-based response will greatly facilitate the development of consortia of probiotics that can be used to enhance animal health and productivity. More importantly, experimental identification of desirable actions by allocthonous bacteria, will allow microbial geneticists to genetically modify strains of such bacteria with amplified capabilities. A sophisticated approach to the formulation of probiotics would allow for the creation of powerful consortia that could, conceivably, prevent or even cure certain diseased states in poultry and other domestic animals. In addition, it is possible that consortia could be developed that would minimize the zoonogens that could impact human health.
Publications
- Croom, W. J., B. M. Berg and I. L. Taylor. 2004 Method of Reducing alumunium levels in the central nervous system. U.S. Patent # 6,734,166 B1
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Progress 10/01/02 to 09/30/03
Outputs
This project was designed to investigate the role of gastrointestinal peptides in maintaining the health and function of the gastrointestinal tract. Furthermore, it seeks to develop a better understanding of the mechanisms of action of probiotics in maintaining and restoring gastrointestinal health. The effectiveness of probiotics is likely dependent on the viability of autochthonous bacteria within the intestinal tract. We have examined the effects of the commonly used ionophore antibiotics, sodium monensin and laidlomycin propionate, all of which have the potential for altering the pattern of microorganisms inhabiting the gastrointestinal tract, on gastrointestinal function using the Swiss-Webster mouse as a model. Of special interest were the effects of these antimicrobials on jejunal glucose function and jejunal and whole-body energetics. Thirty mice (10 mice per treatment) were fed, sodium monensin, laidlomycin or laidlomycin propionate at either, 1.62 or 16.2mM
in the drinking water for 14 days. At the end of the experimental period whole body respiration of each mouse was measured using closed-circuit indirect calorimetry. The mice were then euthanized, jejunal tissue dissected and in vitro jejunal oxygen consumption and glucose uptake were measured. None of the ionophore treatments had any effect on whole body energetics, the rate or efficiency of jejunal glucose absorption or jejunal respiration. It was concluded that ionophore effects on the physiological mechanisms of nutrient absorption was not an issue in animal production. Furthermore, since ionophores can serve as antimicrobials for gram-negative bacteria within the gastrointestinal tract, it was hypothesized, using deductive reasoning, that the autochthonous bacteria within the gastrointestinal tract have little effects on intestinal and whole-body energy metabolism as has been proposed by some investigators. Other projects were initiated to study the effects of direct fed
microbials on small intestinal glucose transport. Of special interest is their effects on insertion of nutrient transporters into the enterocyte membrane since this may well define the limits of maximal intestinal nutrient absorption. Other studies designed to examine the effects of peptideYY on intestinal nutrient transporters in the enterocyte of both turkeys and broilers of different genetic stock have commenced. This laboratory was the first to report the nutrient enhancing effects of peptideYY in the intestinal tract. Recently, investigators at other institutions have reported that another gastrointestinal peptide, epidermal growth factor, increases the insertion of nutrient transporters into the membrane of enterocytes. Unlike epidermal growth factor, peptide YY increases nutrient uptake without concomitant increases in intestinal energy expenditure. We hypothesize that this difference is the result of more efficient insertion of pre-formed nutrient transporters lying within
vesicles just under the enterocyte membrane.
Impacts
The findings of this project could have profound effects on feeding protocols in poultry and other animal species. First, our laboratory is one of the few that understands the significance of changes, in gastrointestinal microflora and the rate and total amounts of nutrient absorption, on whole-body energy expenditures. Our work on probiotics is designed to further the understanding of the physiological mechanisms that result in beneficial effects of probiotics and direct-fed microbials on nutrient absorption. It is hoped that this information can be used to establish a rational approach to the development of probiotics that do not have a deleterious effect on whole-body energetics for use in the poultry industry. Our previous advances in designing technology to enhance intestinal nutrient absorption in poultry (peptideYY administration) represent a quantum leap in the quest of the poultry industry to minimize nutrient input and maximize growth. The work currently in
progress will clarify the changes in physiological mechanisms associated with this increase and identify any differences associated with genotype that may be of use to the poultry industry.
Publications
- Chang, S. C., M. J. Lin, J. Croom and Y. K. Fan. 2003. Administration of triiodothyronine and dopamine to broiler chicks increases growth, feed conversion and visceral organ mass. Poult. Sci. 82: 285-293.
- Fan, Y. K., J. Croom, E. J. Eisen, H. R. Spires and L. R. Daniel. 2003. Ionophores have limited effects on jejunal glucose absorption and energy metabolism in mice. J. Anim. Sci. 81: 2072-2079.
- Korchazhkina, O. V., A. E. Ascroft, J. Croom and C. Exley. 2003. Does either the gastrointestinal peptide PYY or the neuropeptide NPY bind aluminum? J. Inorg. Biochem. 94:372-380.
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Progress 10/01/01 to 09/30/02
Outputs
Phase 1 of the project was devoted to the continuing characterization of the biological effects of the gastrointestinal peptide, peptide YY (PYY), on intestinal function as well as potential regulatory functions in the central nervous system. We were able to conclude that in order to obtain increases in intestinal glucose absorption in hatchling turkey poults via in ovo administration of PYY, careful consideration must be given to the genotype of the fertile egg to which the peptide is administered. In general, in order to increase post-hatch intestinal glucose absorption in poults via in ovo administration, higher dosages of PYY must be administered to eggs from lines selected for growth as opposed to lines selected for egg production. We also concluded that the variation in response to this technology between lines may not be solely due to the genotypic differences in the embryos, but may also be due to differences in the structure of embryonic membranes. The
kinetics of absorption of small peptides into the allantoin is poorly understood. Additionally, embryos are capable of ingesting amnionic fluid as early as day 10 of incubation. Hence, any line differences in the ability of PYY to cross embryonic membranes may significantly affect post-hatch glucose absorption. We were able to determine that no effect is elicited with PYY in any line at dosage levels below 600 micrograms per kilogram of egg weight. We have also demonstrated that administration of in ovo PYY has no detrimental effect on whole-body energy metabolism, as measured by indirect calorimetry, on growing turkey poults through 3 weeks post-hatch. In an closely related paper conducted in cooperation with the Tulane University School of Medicine, we have speculated that PYY may actually increase the efficiency of nutrient utilization in humans to such an extent as to be one of the factors involved with the onset of obesity. Phase 2 of the project was designed to investigate the
effects of administration of probiotics on the absorptive function of jejunal tissue in broilers and poults. Of special interest was the effects of surface nutrient transporters, since probiotics bind to the surface of enterocytes. We started by addressing a very fundamental issue raised by Illinois investigators regarding probiotics, that is their effect on energy metabolism. For this we used the mouse as a model because of our ability to monitor the feed intake and water consumption of large numbers of individual mice. Our studies have shown that administration of ionophore antibiotics with a gram negative spectrum of activity had no effect on jejunal of or whole body energy metabolism as measured by oxygen consumption and indirect calorimetry, respectively, compared to controls not receiving antibiotics. This indirectly demonstrates that speculation by Illinois investigators that both non- pathogenic allocthonous and autocthonous populations of intestinal microorganisms exert
little effect on intestinal tissue and whole body energy metabolism is probably not correct. Further studies are planned using antibiotics with a wider spectra of antimicrobial activity.
Impacts
Our project clearly shows that in ovo administration of select gastrointestinal peptides has the potential for improving the health and productivity of poultry. We have determined, however, that close attention must be given to the strain of poultry used in regards to in ovo dosage. These findings should hasten the development of this technology for use in the poultry industry. In addition, we have found that the use of probiotics does not seem to effect tissue or whole bird energy consumption. This is an important finding given the increased use of probiotics to replace antimicrobials in poultry production systems. Again, this should strengthen the case for the use of safe probiotic technology as a substitute for antibiotics in poultry prodution systems.
Publications
- Bird, A. R., W. J. Croom, Jr. and B. W. McBride. 2002. Dietary management of the intestinal microbiota using probiotics and prebiotics in humans and animals. Proc. Aust. Poult. Sci. Sym. 14:35-43
- Taylor, I. L. and J. Croom. 2002. Enhanced efficiency of nutrient absorption may play a role in obesity: Role of peptide YY. in: Gut-Brain Peptides in the New Millennium,pp. 335-341. (eds) Y. Tache. CURE Foundation, Los Angeles, CA.
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Progress 10/01/00 to 09/30/01
Outputs
Investigations were continued to determine the types and sources of variation in intestinal absorptive response to the in ovo administration of epidermal growth factor (EGF) and peptide YY (PYY) in poultry. Additonally, studies were continued on the mechanisms of action of these gastrointestinal peptides on enterocytes and the possible involvement of the brain-gut axis in their effects. Dosage, genotype and a dosage x genotype interaction have been identified as key sources of variantion. Egg Line and Nicholas poults tend to be more responsive to administration of in ovo PYY than BUT poults in regards to enhancement of jejunal glucose uptake. Corollary studies in my laboratory have shown that subcutaneous PYY administration in control and Down syndrome mice results in rapid mobilization of the brain aluminum pool over a three day period. Aluminum is ubiquitous in our environment including our food and feed as well as water and air. Aluminum inhibits collagen synthesis
in chicken fibroblasts. In humans, aluminum has been been linked as a possible etiological factor for Alzheimer's disease and may depress appetite. Hence, PYY adminsitration may have other benerfical effects in both poultry and humans because of possible involvement of the brain-gut axis in its response. Further studies are underway to repeat and clarify this unexpected action of PYY. These beneficial responses may include enhanced rate of growth in poultry due to greater connective tissue synthesis and increase appetite. In humans, this response may result in a treatment or cure for Alzheimer's disease.
Impacts
In poultry, our preliminary data suggest that both in ovo and post-hatch adminsitration of gastrointestinal peptides, especially epidermal growth factor and peptide YY, may result in less enteric disease and increased performance in growing chickens and poultry, especially during the period of early growth when their gastrointestinal tracts are more immature. In humans, PYY may be used to enhance the absorption of nutrients in patients suffering from degenerative diseases. Additionally,since it has already been safely administered i.v. to humans, it may evolve into a treatment or cure for Alzheimer's disease.
Publications
- Coles, B.A., J. Croom, L. R. Daniel and V. L. Christensen.2001. In ovo peptide YY administration improves body weight at hatch and day 3 in turkey poults. J. Applied Poul. Res. 10:380.
- Croom, J. Piracetam study: Poorly designed and misinterpreted.2001. Arch. Pediatr. Adolesc. Med. 55:1176
- Croom, J. and I. L. Taylor.2001. Neuropeptide Y, peptide YY and aluminum in Alzheimer's disease: Is there an etiological relationship? J. Inorganic Biochem. 87:51
- Peebles, E. D., J. Croom, W. R. Maslin, S. K. Whitmarsh, L. R. Daniel and I. L. Taylor. 2001. In ovo peptide YY administration and epidermal growth factor administration and their effects on growth, yolk sac absorption and yolk stalk function in post hatch broiler chicks. Comp. Physiol. Biochem. Part A, 130:741
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Progress 10/01/99 to 09/30/00
Outputs
Investigations were conducted to determine the effects of in ovo administration of the gastrointestinal peptide, peptideYY (PYY), on the ontogeny, rate and efficiency of glucose transport in the intestinal tract of turkeys. Unlike earlier studies, in ovo administration of 400 micrograms PYY/kg egg weight did not affect the ontogeny of jejunal glucose uptake three days later when administered at days 20, 23 and 26 of incubation. It was noted that the apparent energetic efficiency (APEE) of jejunal glucose absorption increased (represented by a numerical decrease in APEE) between day 26 of incubation and hatch in embryos and poults from saline treated eggs. Administration of PYY at days 23 and 26 of incubation resulted in a decrease (represented by a numerical increase in APEE) in efficiency of glucose absorption at day 26 of incubation and hatch, respectively. Additionally, PYY administration increased jejunal weight adjusted for body weight and decreased jejunal
length adjusted for body weight. It is possible that a response in jejunal glucose transport was not observed with PYY because of a recently identified dosage X genotype interaction with turkeys. The dosage used in the present study was below that (900 micrograms/kg egg weight) known to stimulate jejunal glucose absorption in BUT poults. Ultrastructural investigations into the possible in ovo mechanisms of action of PYY, epidermal growth factor (EGF) and thyroxine on intestinal function indicate both EGF and thyroxine stimulated terminal web development in BUT turkey poult jejunal enterocytes at hatch when administered at d25 of incubation. No differences in enterocyte terminal web development were noted in poults from control and PYY treated eggs. This data suggest that PYY enhances glucose absorption via different cellular mechanisms than EGF or thyroxine. Further studies are underway to more fully characterize the mechanisms of action of in ovo administered PYY in turkey poults
using quantitative Western Blot technology.
Impacts
The development of new technologies, such as in ovo peptideYY administration, that increase the amount and efficiency of nutrient absorption by poultry will favorably impact the nation's poultry industry. Increasing the rate and efficiency of nutrient absorption in broilers and turkey poults will decrease the amount of feedstuffs required to bring birds to market as well as decrease manure output, which will increase industry profits and decrease the impact of the industry on the environment.
Publications
- Berg, B.M., Croom, J., Fernanez, J.M., Spears, J.W., Eisen, E.J., Taylor, I.L., Daniel, L.R., Coles, B.A., Boeheim, F., and Mannon, P.J.2000. Peptide YY administration decreases brain aluminum in the Ts65Dn Down syndrome mouse model. Growth, Development and Aging. 64:3-19.
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Progress 01/01/98 to 12/31/98
Outputs
Studies were conducted to further investigate the effects of in ovo administration of peptide YY (PYY) on growth and feed conversion ratios in broiler chicks and intestinal glucose uptake rate in hatchling turkey poults. In the first study, 1260 broiler chicken eggs were injected at 18 days of incubation and 7 days post-hatch with either saline (control) or 600 mcg PYY/kg egg weight. Birds were housed by sex and treatment in pens of 20 and weekly pen weights and feed intake were recorded. Although no differences were noted in chick performance at 7 days, there was significant (p<. 08) enhancement of feed conversion ratios at 21 days post hatch and body weight (p<. 10) at 42 days. Alterations in the length of incubation time by in ovo PYY administration or a delay in the placement of chicks may have masked beneficial effects of PYY during the early post-hatch period. In a second study, the ability of in ovo administration of epidermal growth factor and PYY to enhance
intestinal glucose transport in hatchling poults from three commercial turkey lines (1 egg line and 2 growth lines) was investigated. In ovo administration of EGF did not increase intestinal glucose transport in any of the three lines studied. In ovo PYY administration (600 mcg PYY/kg egg weight) increased glucose transport in the egg line and one growth line. The second growth line did not show a response to PYY until the dosage was raised to 900 mcg/kg egg weight. These data suggest that genotype may play an important role in the response to PYY.
Impacts
(N/A)
Publications
- Croom, W.J., McBride, B., Bird, A.R., Fan, Y.K., Odle, J., Froetschel, M., and Taylor, I.L. 1998. Regulation of intestinal glucose absorption: A new issue in animal science. Can. J. Anim. Sci. 78:1-13.
- Fan, Y.K., Croom, W.J.,Jr., Christensen, V.L., Bird, A.R., Daniel, L.R., McBride, B.W., Eisen, E.J. 1998. Apparent energetic efficiency of jejunal glucose uptake in young adult turkeys selected for rapid growth. Can. J. Anim. Sci. 78:301-306.
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Progress 01/01/97 to 12/31/97
Outputs
Studies were conducted to investigate the effects of in ovo administration of peptide YY (PYY) on intestinal glucose (IG) uptake rate in hatchling turkey poults and growth and feed conversion (FC) ratios in broiler chickens. In the first experiment, 600 broiler eggs were injected in the air cell at day 18 of incubation with either saline (control) or saline plus 600 mcg of PYY/kg of egg weight. At hatching, 210 chicks from control and 210 chicks from PYY treated eggs were divided into groups of 7, by treatment, in a randomized complete block design. Birds were fed a starter diet from 0-21 days and a grower diet from 21-42 days of age. Body weights were measured at day 1, 7, 21 and 42 posthatch. Feed consumption (FC) was measured weekly by pen. Chick body wt and adjusted pen FC ratio were improved at day 7 by in ovo treatment with PYY (165.7g Vs 170.2g, p<.02 and 1.55 Vs 1.49, p<.04 for control and PYY treatments, respectively). In a second study, turkey eggs were
injected (via the air cell) at day 25 of incubation with either physiological saline or saline plus 600 micrograms of PYY/kg of egg wt. In ovo treatment resulted in increases in IG uptake in hatchling turkey poults. Several studies indicate that in ovo treatment of eggs from both egg-line and growth-line chickens and turkeys increases IG uptake in hatchlings. Thus, in ovo treatment of IG peptides of endogenous origin may enhance chick and poult survival and subsequent performance.
Impacts
(N/A)
Publications
- uptake and oxygen consumption in turkey poults selected for rapid
- growth. Poultry Sci. 76:1739-1745.
- FAN, Y.K., CROOM, J., CHRISTENSEN, V.L., BLACK, B.L., BIRD, A.R.,
- DANIEL, L.R., MCBRIDE, B.W. and EISEN, E.J. 1997. Jejunal glucose
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