Animal & Avian Sciences Theses and Dissertations
Permanent URI for this collectionhttp://hdl.handle.net/1903/2741
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Item CAUSATIVE AGENTS FOR FOWL TYPHOID AND PULLORUM DISEASE IN POULTRY AND APPROACH TO CONTROL(2023) Julianingsih, Dita; Biswas, Debabrata; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)In response to escalating consumer demand, a significant number of conventional US poultry farms have transitioned to antibiotic-free and chemical-free practices, particularly those adopting pasture/organic methods. However, recent reports highlight a resurgence of bacterial diseases in both conventional and pasture poultry farms, resulting in elevated bird mortality rates and reduced profitability. This comprehensive study investigates the prevalence of S. Gallinarum and S. Pullorum, causative agents of fowl typhoid and pullorum disease, in integrated crop-livestock/pasture farm environments and meat products. A total of 1,286 samples from 7 farms and 13 retail markets were examined, revealing that S. Pullorum and S. Gallinarum are common in both farm poultry environments and market products. Antibiotic resistance patterns, determined through an antibiogram assay, indicated high resistance to multiple antibiotics. S. Pullorum and S. Gallinarum were discovered in 2.7% and 1.5% of samples, respectively, at the pre-harvest stage. Only 1.6% of the meat samples recovered from retail markets had S. Gallinarum detected in them at the post-harvest level. Concurrently, a different study investigates the possibility of Orange Cold-press Valencia Terpeneless, a citrus oil variety, acting as a natural antimicrobial in poultry farming. This study tackles the problems caused by a decline in the usage of antibiotics, which has resulted in an increase in bacterial infections. Citrus oil exhibits potential as an antimicrobial agent, inhibiting the growth of S. Pullorum and S. Gallinarum, with consistent MIC and MBC values. Time-dependent experiments with 0.4% citrus oil show total suppression of bacterial growth, which is confirmed by environmental simulations. Furthermore, the study reveals that both Salmonella strains have downregulated their virulence genes, which may indicate a change in the pathogenicity of the bacteria. Overall, the findings highlight the crucial importance of surveillance programs and preventive measures. Citrus oil is presented as a promising natural alternative for antibiotics in the treatment of Salmonella-related infections in the poultry farming industry.Item The study of hyperketonemia in the dairy cow.(2023) Barrientos-Blanco, Mario Alberto; Rico, Eduardo; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)The metabolic phenomenon of ketosis in dairy cows has remained ambiguous, casting uncertainty over our understanding and its real implications. Ketosis, commonly defined as blood β-hydroxy-butyrate (BHB) ≥ 1.2 mM (i.e., hyperketonemia), has been observationally connected to the onset of peripartal metabolic disorders (e.g., infectious diseases, fatty liver), and reduced milk yield in dairy cows. Although BHB is currently used as standard biomarker for the prediction of negative health and performance outcomes during the peripartum, the nature of this relationship is ambiguous. In contraposition, recent discoveries in mammalian biology indicate BHB as therapeutic metabolite (e.g., alleviation of inflammation and oxidative stress). Our overreaching goal was to study the effects of BHB on dairy cow metabolism and health. In our first study, 6 multiparous (parity = 2.8 ± 0.9) Holstein mid-lactation dairy cows (128 ± 52 days in milk; DIM), were enrolled in a study to evaluate a ketogenic diet using calcium butyrate (—CaBu—; a ruminal ketone precursor) against an un-supplemented control (Control) in a crossover arrangement of treatments. The CaBu resulted in nutritional ketosis (P < 0.05) with blood BHB levels of 0.2 mM higher relative to Control. Although CaBu resulted in reduced dry matter intake (DMI; P < 0.05), milk production was not affected (P > 0.40), and feed efficiencies were improved (P < 0.05) relative to Control. No differences in glucose, NEFA, respiration rates, pain scores, or rectal temperatures were observed between treatments. In the second experiment, 8 multiparous Holstein (2.75 ± 0.89) mid-lactation dairy cows (140 ± 48 DIM), feed ad libitum, were enrolled in a in a crossover arrangement of treatments. The aim of the study was to evaluate the effect of ketones by intravenous infusion of either Na-BHB solution (2.5mM; EK) to sustain hyperketonemia —BHB > 1.2 mM and < 3.0 mM—, or NaCl as a control (2.5mM; Control) over a 72h period. A systemic lipopolysaccharide (LPS) challenge (E. coli 055:B5; 0,085 g/kg BW,) was intravenously administered at h 60 from infusion start. Cows sustained hyperketonemia throughout the 72h experimental period (1.4 BHB mM vs. 0.7 BHB mM in EK vs. Control, respectively). While DMI and milk production were not affected by the BHB infusion, the combination with the LPS challenge resulted in reductions of 20.8% (P < 0.05) and 40.1%, (P = 0.14) for both measurements in EK vs. Control, respectively. No differences were detected in the glucose and NEFA concentrations, but insulin was higher 46.6% (P < 0.05) in EK group. Among the immune markers, IL-1 was 30.8% higher (P < 0.05) in the EK group, and not differences were detected in TNF, IL-10, CRP, and caspase-1. As expected, the LPS challenge induced increased respiration rates, temperature, and pain scores over the time course of the evaluation (P < 0.001); however, respiration rates tended to be reduced in 8.4% (P < 0.1) and rectal temperature increased in 0.3% (P < 0.05) by the BHB treatment (P < 0.05). Our results are indicative that, in the absence of an immune challenge, hyperketonemia results in no negative impact on cow productivity and health. These data add support to our hypothesis that cofactors other than ketones may be necessary for the development of negative trajectories of health and performance of lactating dairy cows. Future studies will be required to confirm that BHB hyperketonemia metabolic effects could differ from ketosis disorder in dairy cows.Item In Vitro Study of an Orange Oil Derived Alternative to Antibiotics in the Treatment of Bovine Mastitis(2015) Federman, Cassandra Skye; Biswas, Debabrata; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Bovine mastitis is a costly disease in the U.S. dairy industry. Its major causative agent Staphylococcus aureus is often unresponsive to antibiotic therapy. Our first study examined terpeneless, cold-pressed Valencia (CPV) orange oil as a possible alternative to antibiotic therapy in the treatment of S. aureus associated bovine mastitis. Orange oil showed significant inhibition of S. aureus growth and invasion of bovine epithelial mammary cells, but only modest reductions in pre-formed biofilms, which contribute to persistence of S. aureus infections. Our second study examined major components of terpeneless, CPV orange oil. Of four major compounds tested, only citral and linalool had significant inhibitory effects on S. aureus growth. In addition, they were capable of reducing pre-formed biofilms as well as association and invasion to bovine epithelial mammary cells. Part of this inhibition was due to downregulation of virulence and biofilm genes.Item Lineage reprogramming of tumor-infiltrating cytotoxic T lymphocytes using protein stem cell transcription factors(2015) BhaduriHauck, Anjuli Lucija; Xiao, Zhengguo; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Adoptive cell transfer therapy (ACT) is one of the most promising immunotherapies against cancer. However, this treatment regimen requires the expansion of a small population of effector cells, known as tumor infiltrating lymphocytes, into the billions in order to overcome the immunosuppressive tumor microenvironment. The cytotoxic T lymphocytes (CTLs) within this invading immune cell population are the most critical components to kill the growing cancer cells. Nonetheless, the rapid expansion of already exhausted tumor-infiltrating cytotoxic T lymphocytes (TICTLs) may further push them to a terminally differentiated state that reduces their proliferative response upon antigen stimulation. Recently, induced pluripotent stem cells (iPSCs) generated from TICTLs have been suggested as a way to create a renewable source of rejuvenated tumor-specific CTLs, but retroviral reprogramming is inefficient, and can lead to an increased chance of tumorigenesis. To improve the expansion of TICTLs, we used transient protein exposure to SOX2, OCT4, and NANOG (SON) in order to push these exhausted TICTLs to a less differentiated stage, preferably stem cell-like memory CTLs (Tscm). These three transcription factors were transiently delivered using a nuclear protein delivery system. We found only the TICTLs treated with SON (STICTLs) exhibited an increased proliferation rate and extended survivability, independent of additional cytokines and antigen stimulation both in vitro and in vivo; effector CTLs did not respond to the SON regimen. These highly proliferative STICTLs could be associated with up regulation of certain genes related in cell cycle control, such as cyclin D1. Though these STICTLs still express a T cell receptor (TCR), as well as many critical downstream components, they were unable to elicit a reaction against antigen exposure. Though clearly not iPSCs, it is possible that the SON treatment had pushed the TICTLs into a state similar to an early double negative thymocyte. Our findings indicate that TICTLs are uniquely responsive to protein SON compared to naïve and effector CTLs; suggesting TICTLs may also be sensitive to regulation by other more lineage specific transcription factors, thus present new avenue for cancer immune therapy.Item The role of interleukin-12 for mTOR regulation of memory CTLs(2014) Garcia, Karla; Xiao, Zhengguo; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)A major goal of vaccines is to induce functional immune memory, and efforts to improve the efficacy of vaccines targeting memory CTLs have revealed an important immunoregulatory role of rapamycin, a specific mTOR inhibitor. While inflammatory cytokines are critical for memory CTLs formation, it is unknown if cytokines such as IL-12 mediate rapamycin's regulation during infection. Inhibition of mTOR by rapamycin represses CTL expansion but enhances central memory during vaccinia virus infection in mice. Without IL-12, immunoregulatory effects of rapamycin on CTL expansion and subsequent memory formation are diminished, yet present compared to CTLs not treated with rapamycin. In infected mice, rapamycin directly enhances IL-12 signaling in WT CTLs by upregulating IL-12 receptor-B2 and STAT4 phosphorylation. Furthermore, secondary expansion of rapamycin-regulated memory CTLs in IL-12 receptor knockouts is impaired and resultant secondary memory CTLs are abolished. This indicates that interplay between cytokines and adjuvants should be considered during vaccine design.Item Identification and characterization of long intergenic noncoding RNAs associated with Marek's disease resistance in chicken(2013) Zhan, Fei; Song, Jiuzhou; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Marek's disease (MD) is a highly contagious lymphomatous disease of chicken caused by Marek's disease virus (MDV). MDV has steadily evolved toward increased resistance and virulence over the past decades. A promising strategy for MD prevention and control would be the enhancement of genetic resistance. This study aimed to investigate the roles of long intergenic noncoding RNAs (lincRNAs) in MD resistance and susceptibility in chickens. We reported more than 1000 lincRNA loci in chicken. Computational functional annotation suggested that lincRNAs were involved in a wide range of biological processes. Moreover, distinct lincRNA expression signatures were observed between MD resistance and susceptible chickens. Additionally, a candidate lincRNA termed linc-stab1 was identified and it may play an important role in MD immune response by regulating a nearby protein-coding gene STAB1. In summary, our results demonstrated that lincRNAs also play an important role in MD resistance and provide good candidates for hypothesis-driven functional studies.Item GENOME-WIDE ANALYSIS OF CHICKEN MIRNAS AND DNA METHYLATION AND THEIR ROLES IN MAREK'S DISEASE RESISTANCE AND SUSCEPTIBILITY(2012) Tian, Fei; Song, Jiuzhou; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Marek's disease (MD) is a T cell lymphoma in chickens and causes high mortality and morbidity in productive chickens. Two inbred chicken lines, resistant line 63 and susceptible line 72, with the same MHC haplotype, showed distinct disease outcomes after MDV infection. The current studies aimed to illustrate the role of microRNA (miRNAs) and DNA methylation in MD resistance and susceptibility in chickens. First, to ascertain the function of miRNAs, miRNA microarray experiments were used to identify miRNAs sensitive to MDV infection in the 2 lines. Most miRNAs were repressed in line 72 after MDV infection, while their transcription was steady in line 63. The miRNA target genes were identified in chickens. Cellular miRNA gga-miR-15b and gga-let-7iwere reduced in infected line 72 chickens and MD tumors. The downregulation of the two miRNAs increased the expression of ATF2 (activating transcription factor 2) and DNMT3a (DNA methyltransferase 3a) in infected line 72. These results indicated that miRNAs may play antiviral functions through modulating target gene expression. Next, to characterize the role of miRNAs in MDV infection, the selected chicken miRNAs were overexpressed in MDV infected DF-1 cells. The overexpressions of chicken miRNA gga-miR-15b and gga-let-7i, by using the retroviral based vector, significantly restricted MDV replications in vitro. MDV oncoprotein was repressed, suggesting that chicken miRNAs may limit MDV propagation. Finally, we found deregulation of transcription of DNA methyltransfereases (DNMTs) in lines 63 and 72 after MDV infection, which coordinated with the methylation alterations in the 2 lines. Infection induced differential methylation regions (iDMRs) that were identified through genome-wide DNA methylation quantification. Genes overlapping line-specific iDMRs were related with pathways of different functions in these two lines, implying the involvement of DNA methylation in MD- resistance and susceptibility. An in vitro study showed that DNA methylation inhibitor repressed viral spread and viral replication. In conclusion, the observed variations of miRNA expression and DNA methylation may be associated with disease predisposition in chickens.Item Generating memory cytotoxic T lymphocytes through repetitive peptide boosting(2012) Smyth, Kendra; Xiao, Zhengguo; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Cytotoxic T lymphocytes (CTLs) play a critical role in controlling intracellular pathogens and cancer cells, and induction of memory CTLs holds promise for developing effective vaccines against critical virus infections. However, generating memory CTLs remains a major challenge for conventional vector-based, prime-boost vaccinations. Thus, it is imperative that we explore nonconventional alternatives, such as boosting without vectors. We show here that repetitive intravenous boosting with peptide and adjuvant generates memory CD8 T cells of sufficient quality and quantity to protect against infection in mice. The resulting memory CTLs possess a unique and long-lasting effector memory phenotype, characterized by decreased interferon-gamma but increased granzyme B production. These results are independent of the specific adjuvant applied and are observed in both transgenic and endogenous models. Overall, our findings have important implications for future vaccine development, as they suggest that intravenous peptide boosting with adjuvant following priming can induce long-term functional memory CTLs.