Master of Science in Biological Sciences, Youngstown State University, 2015, Department of Biological Sciences and Chemistry

The human microbiome is home to roughly 100 trillion microbial, which have been shown to assist in various host physiological functions. Research shows that probiotic consumption may benefit host health. Therefore this is a two part study: project I is a study determining the effects of yogurt (probiotic) consumption on GI microbial structure and diversity, and project II is a study examining the effects of probiotic pill consumption has on stress response (cortisol) for healthy human test subjects exposed to a stressor. In chapter II we used terminal restriction fragment length polymorphism, Sanger sequencing, and qPCR to characterize microbial communities of the GI tract. Non-metric multidimensional scaling analyses, Shannon Wiener diversity index values, and qPCR (targeting Lactobacillus) analysis showed a temporal influence on microbial community, diversity and absolute abundance of Lactobacillus on human test subjects that consumed 250g of yogurt per day for 42 consecutive days. In chapter III, we quantitated Lactobacillus using qPCR and determined stress response (via cortisol) using a salivary enzyme immune assay (EIA). Results from this study showed a decrease in stress response and an increase in the relative abundance of Lactobacillus in individuals that consumed one probiotic pill per day for 30 consecutive days. In comparison, the control group showed no changes in stress response or Lactobacillus. Results from this study suggest the potential for probiotics as therapeutic treatment for host physiological function and stress response. However, further research is necessary to determine the effects probiotics have on human test subject.

Committee: Carl Johnston PhD (Advisor); Diana Fagan PhD (Committee Member); Xiangjia Min PhD (Committee Member) Subjects: Biology; Microbiology; Molecular Biology

Doctor of Philosophy, The Ohio State University, 2024, Animal Sciences

Salmonella, a foodborne zoonotic pathogen, is the leading cause of death associated with foodborne illness in the USA. Salmonellosis is primarily transmitted through the consumption of contaminated poultry meat and eggs. The irrational use of antibiotics to control Salmonella infection in poultry has led to the emergence of multidrug-resistant (MDR) strains and its horizontal transmission to humans. Additionally, vaccines used to prevent Salmonella infection do not provide cross-protection to heterologous serotypes of Salmonella. Therefore, there is an urgency of the development of novel antimicrobials to control Salmonella infection. Studies have shown that small specialty crop farms (SSCFs), which practices mixed farming, are reservoirs of MDR Salmonella. Regular monitoring is needed to reduce the risk of MDR Salmonella transmission from SSCFs to humans. To this end, we isolated Salmonella from 29 of 1174 environmental samples such as dairy and poultry manure, soil, and water of SSCFs. From 29 positive samples, 80 isolates were subjected to whole genome sequencing (WGS) to determine the serotypes, antimicrobial resistance genes, virulence genes, plasmids to understand the potential risk of its transmission. Our results detected the presence of ACSSuT cassette in six Salmonella isolates, which confers resistance to ampicillin, chloramphenicol, sulfonamide, streptomycin, and tetracycline, demonstrating the urgency to develop the novel antimicrobials to combat MDR Salmonella. Therefore, we assessed the activity of novel antibiotics alternatives such as probiotics and anti-microbial peptides (AMPs) against Salmonella. The anti-Salmonella activity of probiotics and AMPs characterized in our study could lay foundation as novel antimicrobials against Salmonella. In silico serotyping of Salmonella isolated from SSCFs identified 15 serotypes of Salmonella. Eight out of 15 serotypes reported in our study are among the top 20 serotypes frequently reported in human salmone (open full item for complete abstract)

Committee: Gireesh Rajashekara (Advisor) Subjects: Biology

Master of Science, The Ohio State University, 2024, Animal Sciences

In the swine industry, weaning is a critical period due to changes in the environment and disruption of feed intake. In neonatal pigs, the gastrointestinal tract (GIT) is still developing and relies on the diet to provide nutrients for growth and development. The lack of nutrients hinders GIT growth and development leading to villus atrophy. As a result, the GIT cannot restore and maintain normal physiological functions because nutrients from the diet cannot be effectively absorbed and utilized. Previously, antibiotics were the standard to resolve the negative effects of weaning including disruption of feed intake. However, the concern about antibiotic resistance has elicited the use of alternative methods to mitigate those negative effects. Lactic acid bacteria have been recognized to possess probiotic properties and modulate host physiology. Probiotics are used as direct-fed microbials in the swine industry to optimize pig performance and production efficiency. In the nursery, probiotics have been shown to mitigate post-weaning diarrhea caused by enterotoxigenic Escherichia Coli (ETEC) and modulate health parameters, but the data are inconsistent. Microbial genome sequencing could help better define the mechanisms of probiotic mode of action and dose required to improve probiotic efficacy. The experiments conducted for this thesis aimed to determine if LAB Pediococcus acidilactici PECh3A, which has been genome sequenced for determining gut physiology mechanisms, expresses probiotic properties in vitro by conducting dose response analysis and investigate interactions with intestinal porcine enterocyte jejunum cells (IPEC-J2) and ETEC (3030-2: K88ac LT and STb). A growth curve of P. acidilactici was conducted to determine colony-forming units (CFU)/mL concentrations over a 15-hour growth period. Based on the results of the growth curve a dose-response analysis was investigated on concentrations; 108, 109, and 1010 CFU/mL across different probiotic property test (open full item for complete abstract)

Committee: Sheila Jacobi (Advisor); Alejandro Relling (Committee Member); Benjamin Enger (Committee Member) Subjects: Animal Sciences

Doctor of Philosophy, The Ohio State University, 2024, Food Science and Technology

Antibiotics are powerful tools to treat bacterial infections. However, the surge of antibiotic resistance (AR) is considered one of the top threats to food safety and global public health. Every year, close to 5 million deaths are associated with AR bacterial infections with around 1.27 million deaths directly attributed to AR bacterial infections worldwide. Besides the public health threat, around $60-100 trillion financial loss is expected by 2050 if no action is taken. AR commensals, while usually not directly causing death, can disseminate AR genes to pathogens, and thus indirectly negate effective therapy, making people vulnerable again. Thus, even the most common AR bacterial infections, like mid-ear infections and tooth extraction, may become life-threatening again. For decades, the overuse of antibiotics has been considered as the main cause of AR. Thus, limiting the use of antibiotics has been the primary control strategy worldwide. However, in reality, AR bacteria are still prevalent and persistent in human and animal hosts, as well as the environment even without antibiotic exposure. Humans and animals will get sick and inevitably need antibiotic treatment. AR is a complicated issue and there are multiple contributors to it. Besides the applications of antibiotics, it is now recognized that the rich gut microbiota residing in human and animal gastrointestinal tract system as a breeding ground have key roles in the AR ecology from evolution, dissemination, enrichment and persistence. Oral/nasal introduced AR bacteria proliferate and persist in the host gut. Gut-impacting (orally administered and biliary excreted) antibiotics further selectively enrich AR bacteria. The AR-rich human and animal feces have served as the most significant avenue impacting the AR gene pool in the ecosystem. Therefore, an improved understanding of AR persistent mechanisms and effective mitigation of the pre-existing, persistent AR bacteria in host GI tract system are essential fo (open full item for complete abstract)

Committee: Hua Wang (Advisor); Matthias Klein (Committee Member); Zhongtang Yu (Committee Member); Ye Xia (Committee Member); Michael Cressman (Committee Member) Subjects: Food Science; Health

Doctor of Philosophy, The Ohio State University, 2023, Food Science and Technology

Kefir is a fermented dairy beverage with origin in the Caucasus mountains that has been consumed for centuries due to its attributed health benefits, primarily owed to its diverse microbiota and their valuable metabolites. Kefir has been referred to as one of the “9 food trends to watch for in 2021” by the Institute of Food Technologist and consumers are incorporating kefir-products into their diets as a potential probiotic product. However, consumption of traditional or commercial kefir as a probiotic product presents several drawbacks. Traditional kefir is produced by the fermentation of milk with kefir grains as starter culture, a complex microbial community of bacteria and yeast embedded in an exopolysaccharide matrix. However, according to some reports traditional kefir has limited acceptance among western consumers and due to the complexity of its microbiota and production practices, achieving industrial-level production faces several challenges. Furthermore, in the United States, there is no established standard for commercial kefir products. The United States Food and Drug Administration (FDA) considers microorganisms added to products as dietary supplements, and according to the FDA Code of Federal Regulations Title 21, kefir is categorized as a cultured milk that contains aroma and flavor producing microbial cultures. Therefore, under the present regulation, kefir producing companies have the freedom to incorporate microorganisms without undergoing verification or regulation. As a result, kefir-products on the market are fermented milk products containing different combinations of starter cultures and probiotic strains, varying from one company to the other. Additionally, recent reports indicate that these products do not contain any of the core kefir microorganisms present in traditional kefir and there is lack of research to support their health effects. Given the limitation of traditional kefir and current commercial kefir-products, it is imperative to (open full item for complete abstract)

Committee: Valente B. Alvarez (Advisor); Christopher Simons (Committee Member); Ahmed Yousef (Committee Member); Rafael Jimenez Flores (Committee Member) Subjects: Food Science; Microbiology

Master of Science, The Ohio State University, 2022, Animal Sciences

Enterococcal spondylitis (ES) is a clinical syndrome caused by improper colonization of Enterococcus cecorum in the free thoracic vertebra (FTV) and adjacent notarium or synsacrum. Enterococcal spondylitis is a devastating disease that results in lameness which can lead to lowered feed efficiency, decreased weight gain, and mortality. Because of the relatively recent discovery and recognition of the pathogenesis for ES, research models are still under development. Induction of ES through E. cecorum bacterial translocation, which occurs when the intestinal barriers are damaged or weakened by stressors like pathogens, heat, dietary ingredients, etc. may accurately represent production setting conditions. Some diet additives that have been known to have antinutritional effects and cause intestinal barrier damage include rye high in non-starch polysaccharides (NSPs), animal-based proteins, and poor-quality soybean meals (pqSBM). These ingredients may be able to induce translocation of E. cecorum in broilers and lead to cases of ES. Enterococcus cecorum is a normal component of gastrointestinal flora of chickens and has been found to make up the majority of enterococcal and streptococcal flora in older birds. Since E. cecorum is part of normal gastrointestinal (GI) tract, finding methods to prevent translocation to other areas of the body can be challenging. Thus, promoting and maintaining the intestinal barrier is integral. Probiotics have been documented to influence gut barrier integrity via increased tight junction gene expression. Since the gut barrier prevents transfer of potential pathogens like E. cecorum into the blood stream and probiotics have been shown to increase gut barrier integrity, probiotics may be able to prevent instances of ES. Two separate experiments were conducted, a series of 4 in vitro assays were used to determine the effects of two commercial Bacillus sp. probiotics, GutCare® and Ecobiol® on growth of known pathogenic strains of E. cecorum (open full item for complete abstract)

Committee: Lisa Bielke (Advisor); Sara Mastellar (Committee Member); Shelia Jacobi (Committee Member) Subjects: Agriculture; Animal Diseases; Animal Sciences

DNP, Otterbein University, 2022, Nursing

Clostridium difficile (C. difficile) are toxigenic, spore-forming bacteria that live in the gastrointestinal (GI) tract of healthy individuals without harm to the host due to the protection offered by the natural intestinal flora. Antibiotics can disrupt the intestinal flora and create an environment within the GI tract that promotes the infectious growth of C. difficile. C. difficile infections (CDIs) are the most common cause of antibiotic-associated diarrhea, are highly contagious, and can result in an increased risk of morbidities and mortality to individuals affected. Recent studies and literature reviews identified the use of probiotic therapy in patients receiving antibiotics to be an effective prophylactic intervention for decreasing antibiotic-associated CDI occurrence. The purpose of this organizational assessment was to compile organizational data concerning nosocomial CDIs as a prerequisite to future practice changes regarding the use of probiotics for the prevention of nosocomial antibiotic-associated CDIs. Results to be determined upon completion of data collection and analysis. Conclusions & recommendations to be determined upon completion of data collection and analysis. Co-Author: Schwaderer, Paxton

Committee: Dr. Kacy Ballard (Advisor); Dr. Brian Garrett (Committee Member); Stormy Barton (Committee Member); Elizabeth Finke (Committee Member) Subjects: Health; Health Sciences; Medicine; Nursing; Pharmacy Sciences

Master of Science, The Ohio State University, 2021, Animal Sciences

Necrotic enteritis is an enteric disease primarily caused by overgrowth of C. perfringens (CP) in the small intestine following a variety of predisposing factors. The objective of this study was to determine if a novel probiotic showed anti-clostridial effects, survived pelleting temperatures and the varied environment of the gastrointestinal tract (GIT), and if anti-clostridial effects were retained through the GIT. The probiotic was tested against 9 strains of CP to determine overarching anti-clostridial effect. The probiotic suppressed all 9 strains of CP significantly (p-value<0.05) when CP inoculated media was overlayed onto a pre-grown colony of probiotic and zones of inhibition measured. Next, probiotic efficacy was compared against common antibiotics and other bacterial isolates. Two of the four antibiotic treatments, penicillin (0.0625mg/mL) and metronidazole (0.05mg/mL), two of the bacterial isolates, and the candidate probiotic were all able to reduce CP growth, with the candidate probiotic outperforming the bacterial isolates (p-value<0.001) and metronidazole (p-value=0.007). The CP strain showed resistance to the other two antibiotic treatments, BMD (0.022mg/mL) and avilamycin (0.05mg/mL). A germination and sporulation assay was completed to ensure spores could germinate at the internal body temperature of chickens and sporulate to survive pelleting. A lack of significant change (p-value=0.096) in cell recovery was indicative of the probiotic's ability to germinate to colonize the gut and sporulate to endure pelleting. Interaction of the probiotic candidate with 18 other common enteric residents of the microbiome in poultry were observed in a streak plate assay, in search of potential inhibition. Of the 18 additional strains tested against the probiotic, three strains of E. cecorum were significantly inhibited by the probiotic based on Chi2 analysis. A gastrointestinal survival assay was performed mimicking the crop, proventriculus, and intestines to (open full item for complete abstract)

Committee: Lisa Bielke (Advisor); Zhongtang Yu (Committee Member); Hongyu Xue (Committee Member) Subjects: Animal Sciences

Doctor of Philosophy, The Ohio State University, 2021, Food Science and Technology

Salmonella and Campylobacter are leading bacterial agents both nationally and globally, making them high public health concerns. Both are significant important zoonotic pathogens commonly found in livestock. Poultry and poultry products are the most common sources of human infections. Infected poultry show little to no clinical signs, and risk the chance of entering the food system, potentially contaminating consumers. Direct links between the high load of bacteria in the chicken intestinal tract and the high contamination of poultry carcasses support the need for pre-harvest control. Previously, antibiotics were used to control bacterial infections and growth; but increasing occurrence of antibiotic resistant bacteria has caused laws and practices to shift. Targeting the control of foodborne pathogens in the pre-harvest stage can improve animal welfare and public health. Alternative treatment methods are needed to combat Salmonella and Campylobacter in production animals, improve antibiotic stewardship, and subsequently strengthen the economy. We discovered the antimicrobial efficacy of Lactobacillus acidophilus (LA), Lactobacillus rhamnosus GG (LGG), and Bifidobacterium animalis subsp. lactis (Bb12) in vitro. More importantly, we showed LGG significantly reduced Salmonella in the chicken cecum by 1.9 logs (P<0.001) at 10 days post infection. LGG was also able to inhibit the growth/ presence of other food safety significant Salmonella serovars in an agar well diffusion assay. Characterization of antibacterial activity of LGG revealed that although organic acids are present, the most crucial part of inhibition are the heat and protease stable peptides that were identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). After which, we tested the efficacy of Lactobacillus rhamnosus GG (LGG) derived small peptides (P1-NPSRQERR, P2- PDENK, and P4-MLNERVK) against Salmonella Typhimurium (ST) in chickens and examined their antibacterial effects a (open full item for complete abstract)

Committee: Gireesh Rajashekara (Advisor); Anastasia Vlasova (Committee Member); Melvin Pascall (Committee Member); Scott Kenney (Committee Member) Subjects: Food Science; Microbiology

Master of Sciences, Case Western Reserve University, 2020, Biology

The “mycobiome" refers to the composition of both bacterial and fungal communities in the human gut microbiome and has been the focus of disease-state correlations investigated by researchers and pursued with commercial interests by biotech startups. A microbiome startup currently sells direct-to-consumer at-home microbiome sequencing kits and probiotics that aim to balance the gut biofilm that contributes to the dysbiosis-associated conditions. This company has expressed an interest in developing new business strategies to leverage their intellectual and technical strengths. This thesis is composed of two parts: The first section is a scientific and technical investigation of the micro- and myco-biome, sequencing techniques and strategies (16S, ITS, WGS, and Shotgun Metagenomic Sequencing) that play a role in the characterization and identification of fungal and bacterial colonies in the gut. These strategies aim to overcome challenges in characterizing and quantifying microbiota composition. Next, this sequencing data can form a robust database of patient data that plays a role in disease identification, and this thesis identifies some of the bioinformatic analyses to achieve this goal. The section concludes with how insights derived from patient data can be used in the optimization of cohort design in clinical trials for various diseases. The second section investigates three different business models that a microbiome startup has expressed interest in exploring for future development: (1) medical foods; (2) a therapeutic pipeline; and (3) a data-licensing and discovery platform for drug development. A detailed analysis of the market dynamics, competitive landscape, regulatory issues, and other nascent concerns was performed for each potential vertical as a foundation to develop future business strategy of a microbiome-related startup. The thesis is concluded on a holistic analysis of the scientific and technical assets and business opportunities and str (open full item for complete abstract)

Committee: Christopher Cullis (Committee Member); Emmitt Jolly (Committee Member); Neema Mayhugh (Committee Member) Subjects: Biology; Entrepreneurship

Master of Science, The Ohio State University, 2016, Animal Sciences

This project studied 1. Inhibitory effects of probiotic culture supernatant from four different probiotic strains against Salmonella enterica Enteritidis (S. Enteritidis) in vitro, 2. the effect of a synbiotic product on layer production parameters, gut bacterial profile, and immune parameters, pre- and post-Salmonella challenge, and 3. the effect of organic acid plus phytochemical supplement product on layer production parameters with and without Salmonella vaccine. In the first objective, two experiments were conducted to study the inhibitory ability of culture supernatants from four different probiotic bacterial strains on S. Enteritidis growth in vitro. The first experiment studied the effect of probiotic culture supernatant on Salmonella inhibition in liquid media. Culture supernatants from the probiotics L. reuteri, E. faecium, B. animalis, or P. acidilactici probiotics inhibited (P < 0.05) the growth of Salmonella. The second experiment studied the effects of probiotic supernatants on Salmonella inhibition in solid media. The culture supernatant from E. faecium and L. reuteri separately inhibited Salmonella growth efficiently compared to that from B. animalis and P. acidilactici. In the second objective, the effects of a synbiotic containing four live probiotic strains on layer production and local immune parameters following an experimental Salmonella infection was studied. At 18 and 20 wk of age, birds fed synbiotics in both vaccinated and unvaccinated pullets had higher body weight (P < 0.05) than the non synbiotic control group. Birds fed synbiotics had 0.7%, 17.8%, 21.7%, 3%, and 4.2% higher (P < 0.05) hen day egg production (HDEP) at 19, 20, 21, and 23 wk of age, compared to the birds fed no synbiotics, respectively. At 24 wk of age, birds were challenged with S. enterica Enteritidis. Hens fed synbiotics had 3%, 6.7%, 4.3%, 12.5%, and 14.4% higher (P < 0.05) HDEP at 24, 25, 26, 27, and 28 wk of age, compared to the birds fed no synbiotics, re (open full item for complete abstract)

Committee: Ramesh Selvaraj (Advisor); Michael Lilburn (Committee Member); Renukaradhya Gourapura (Committee Member) Subjects: Animal Sciences

Doctor of Philosophy, The Ohio State University, 2015, Comparative and Veterinary Medicine

Lactobacillus spp. have been tested in infants for the prevention or treatment of various enteric conditions. However, to aid in rational strain selection for specific treatments, comprehensive studies are required to delineate and compare the specific molecules and pathways involved in a less complex but biologically relevant model (gnotobiotic pigs). Here we elucidated Lactobacillus rhamnosus (LGG) and L. acidophilus (LA) specific gut transcriptome responses in a monocolonized pig model to simulate responses in newly colonized infants. Whole genome microarray, followed by biological pathway reconstruction, was used to investigate the host-microbe interactions at early (day 1) and later stages (day 7) of colonization. Both LA and LGG modulated common responses related to host metabolism, gut integrity, and immunity, as well as responses unique to each strain in pigs. Our data indicated that probiotic establishment and beneficial effects in the host are guided by: (1) down-regulation or upregulation of immune function-related genes in the early and later stages of colonization, respectively, and (2) alternations in metabolism of small molecules (vitamins and/or minerals) and macromolecules (carbohydrates, proteins, and lipids). Pathways related to immune modulation and carbohydrate metabolism were more affected by LGG, whereas energy and lipid metabolism-related transcriptome responses were prominently modulated by LA. These findings are highly relevant to the improvement of probiotic-based interventions in enteric infections, either to moderate specific intestinal conditions or to enhance vaccine efficacy for enteric infections. Enteric infections are attributed to millions of deaths in infants worldwide annually. Human rotavirus (HRV) is a major cause of viral gastroenteritis in infants that accounts for approximately 440,000 deaths annually worldwide, particularly in developing countries where malnutrition is prevalent. Malnutrition perturbs the infant gut mic (open full item for complete abstract)

Committee: Gireesh Rajashekara Dr (Advisor); Linda J Saif Dr (Committee Member); Michael T Bailey Dr (Committee Member); Jordi B Torrelles Dr (Committee Member) Subjects: Comparative; Developmental Biology; Gynecology; Health; Medicine; Veterinary Services

Master of Science, The Ohio State University, 2015, Animal Sciences

Bacteria play a crucial role in the gastrointestinal health of horses and stress, medication and diet can alter microbial populations leading to decreased health. Recently there has been increased interest in the prophylactic use of probiotic supplements in healthy horses. Therefore, the objective of this research was to evaluate the influence of probiotic supplements in three separate studies. In the first study, seven Quarter Horse mares (10.0 ± 2.0 yr) were randomly assigned to one of two treatments in a crossover design with 49 d experimental periods. All horses received a control diet of 0.5% BW of 12% CP pelleted concentrate with water and mixed grass hay ad libitum (CTL) or the control diet supplemented with 140 g of a probiotic containing 109 cfu per dose of Lactobacillus acidophilus (TRT) intended for humans. Fecal samples were collected on d 0, 14, 21, 28, 35, 42, and 49. In the second study, 12 Quarter Horses (4 geldings, 8 mares; 1.5 ± 0.5 yr) were randomly assigned to one of two treatments. All horses received a control diet of 0.5% BW of 14% CP pelleted concentrate with water and mixed grass hay ad libitum (CTL) or the control diet supplemented with 10 g of a commercial probiotic containing 107 cfu per dose of Lactobacillus acidophilus (TRT) intended for livestock. Fecal samples were collected weekly for 77 d. In the third study, six Miniature Horse geldings (7.5 ± 3.5 yr) were used in a replicated 3 x 3 Latin Square design. Each horse received a control diet of 1.5% BW of a mixed grass hay and water ad libitum and was randomly assigned to one of three treatment groups during each 14 d period: control diet only (CTL), control diet supplemented with 0.11g/kg of BW of a commercial equine probiotic containing 2.0 x 107 cfu/g of Bacillus subtilis (LO) or control diet supplemented with 0.22 g/kg of BW of the commercial equine probiotic (HI). Fecal samples were collected on d 0, 3, 7, 10 and 14 of each period. For each study, fecal samples were pooled by (open full item for complete abstract)

Committee: Kimberly Cole (Advisor); Zhongtang Yu (Committee Member); Pasha Lyvers-Peffer (Committee Member) Subjects: Animal Sciences

MS, University of Cincinnati, 2015, Medicine: Immunology

In the United States, nearly 500,000 infants are born pre-term each year. These infants are at high risk of serious complications, including necrotizing enterocolitis (NEC), sepsis, and death. While hospitalized these infants routinely receive multiple doses of broad-spectrum antibiotics, which can have profound and lasting effects on the composition of the intestinal microbiome. Microbiome composition, in turn, can impact a variety of health outcomes including NEC and sepsis. We were interested in determining how feeding strategies impact microbiome composition in infants receiving large doses of antibiotics. The standard of care in Cincinnati is for hospitalized pre-term infants to receive human milk (either mother's own milk or pasteurized donor milk) for the first 14 days of life. After that point they may receive formula, milk, or a combination of milk and formula. We hypothesized that infants who continue to receive predominantly human milk in week three of life would have greater microbiome diversity characterized by increased Bacteroides and Bifidobacteria and decreased Clostridia. We examined the intestinal microbiome of 71 infants from two Cincinnati NICUs. Our experimental cohort consisted only of infants born <32 weeks gestation, with birth weight between 850 and 1500 grams. All infants were free of congenital abnormalities, did not display overt feeding difficulties, survived free of NEC or sepsis, and had stool samples in weeks one, two, and three of life. We examined the effect of heavy antibiotics (=5 days treatment) and light antibiotics (1-4 days treatment) versus no antibiotics. We then examined the effect of feeding predominantly human milk versus formula in week three of life within each category of antibiotics treatment. Stool samples were collected, 16s rRNA extracted, and microbiome data analyzed for alpha diversity and beta diversity. Microbiome composition was determined by LEfSe. We determined that in all categories, including heavy antibi (open full item for complete abstract)

Committee: Ardythe Morrow Ph.D. (Committee Chair); Simon Hogan Ph.D. (Committee Member); Jonathan Katz Ph.D. (Committee Member) Subjects: Immunology

Master of Science, The Ohio State University, 2014, Animal Sciences

Ohio has a 158 million dollar turkey industry. Gut growth and development occurs first before the turkey can realize its full muscle producing potential. Alternative and adjunctive approaches to decreasing the use of feed grade antibiotics are becoming an important area of research due to increasing consumer and legislative concerns with antibiotic resistance. Probiotics or supplemental dietary commensal microbes is one such potential approach as they can colonize the intestine, particularly in young animals with a relatively naive intestine microbiome. Intestinal mucosa is made up of mucin glycoproteins that play a key role in preventing the attachment and colonization of pathogenic bacteria. These proteins are made up of a protein backbone that is coded for by the MUC gene family. At hatch, the turkey intestine is relatively aseptic and therefore vulnerable to bacterial colonization from both commensal and pathogenic microbes. In this study, we determined the expression of MUC2, which codes for a secretory gel-forming mucin that is predominantly found in the small and large intestine, from immediately post-hatch through day 11 of age in poults fed a conventional starter diet, the starter diet supplemented with two commercial probiotics (A and B), or the starter diet supplemented with a commercial prebiotic. This was done by comparing the MUC2 transcription levels to the transcription level of a housekeeping gene. Multiple potential housekeeping genes were investigated and one, RPS13, was found to be stably expressed across all ages and treatments in the turkey poult. The effects of the supplemented diets on intestinal development were also analyzed. While MUC2 transcription increased with age, there were no significant effects due to diet. The intestinal parameters of villus height, area and crypt depth were all increased with supplementation of probiotic B and the prebiotic.

Committee: Michael Lilburn (Advisor); Macdonald Wick (Committee Member); Joseph Ottobre (Committee Member) Subjects: Animal Sciences

Doctor of Philosophy, Case Western Reserve University, 2007, Pathology

Elevations in the mucosal expression of IFN-γ and the common p40 subunit of IL-12 and IL-23 accompany early colonic inflammation in interleukin-10 deficient (IL-10 ‑/‑) mice. These cytokines later decline while inflammation persists. Changes in the immune response are known to occur as a component of normal aging, and have been shown to affect disease course in multiple systems. I therefore hypothesized that age-associated variations in systemic and mucosal expression of multiple cytokines in the IL-10 ‑/‑mouse reflect underlying age-related alterations in systemic and mucosal immunity. To address this question, IL-10 ‑/‑and wild-type mice were maintained in an ultrabarrier facility or transferred to conventional housing at 3, 12, or 30 weeks of age, with or without pretreatment with probiotic organisms. Weight, stool changes, and histologic features were followed. Lamina propria mononuclear cells were cultured for cytokine analysis by ELISA or multiplex cytokine array. Ultrabarrier-housed IL-10 ‑/‑mice were found to be statistically indistinguishable from wild-type mice by weight, disease activity index, and histologic inflammation. In addition, IL-10 ‑/‑mice, but not wild-type, transferred at 3 weeks gradually develop colitis, reaching significant, sustained maximum by 15 weeks of age. Transfer at 12 weeks induces rapid disease onset in both strains, maximal at 15 weeks of age. Inflammation persists in IL-10 ‑/‑, while wild-types recover. IL-10 ‑/‑and wild-type mice transferred at 30 weeks demonstrate transient diarrhea and weight loss, but no chronic inflammation. Probiotics delay symptom onset only in 12-week-old group. Age-associated fluctuations in systemic and mucosal levels of several cytokines were found in both IL10 ‑/‑and wild-type mice, and expression of many cytokines was elevated even in the absence of overt disease. LPS induced higher levels of IFN-γ from IL-10 ‑/‑ versusWT splenocytes, but IL-17 increaed with TCR stimulation. These experiments demons (open full item for complete abstract)

Committee: Alan Levine (Advisor) Subjects: Health Sciences, Immunology