Browsing by Author "Dalloul, Rami A."
Now showing 1 - 20 of 52
Results Per Page
Sort Options
- The Acute and Chronic Effects of a Cyclic Heat Stress on 24 to 28 Week Old Laying Hens on Performance, Egg Quality, Apparent Metabolizable Energy, and Blood ChemistryBarrett, Nathaniel W. (Virginia Tech, 2016-11-02)Commercial laying hens are heavily selected for increased egg production, but little selection has occurred for resistance to extreme temperatures. The objective of the experiment was to determine the effects of a daily cyclic heat stress (HS) on performance, body temperature, egg quality, nitrogen corrected apparent metabolizable energy, and blood chemistry in laying hens at acute and chronic time points. In total, 407 pure line laying hen pullets (18 wk) were sourced from Hy-Line International and housed individually for the duration of the experiment. Feed intake, egg production, and body weights were significantly decreased while egg weight and feed efficiency increased due to HS exposure. All egg quality parameters except yolk weight were decreased by HS exposure, including Haugh unit, eggshell and albumen weights. Egg yolk weights were increased over the first 2 wk of HS before falling. Blood chemistry was affected by HS resulting in respiratory alkalosis likely caused by increased respiration and evaporative cooling. There was a decrease in the PCO2, an increase in pH and a reduction in iCa in the blood within 4 to 6 hours of HS when compared to pre-HS levels. The data indicate that the hens were negatively affected by HS and the response was different for acute and chronic time points. At the acute time point the changes to the hen's physiology were caused by altered blood chemistry, but after chronic exposure, the low feed intake seemed to influence the responses.
- Antibiotic perturbation of gut bacteria does not significantly alter host responses to ocular disease in a songbird speciesWeitzman, Chava L.; Belden, Lisa K.; May, Meghan; Langager, Marissa M.; Dalloul, Rami A.; Hawley, Dana M. (PeerJ, 2022-06-10)Bacterial communities in and on wild hosts are increasingly appreciated for their importance in host health. Through both direct and indirect interactions, bacteria lining vertebrate gut mucosa provide hosts protection against infectious pathogens, sometimes even in distal body regions through immune regulation. In house finches (Haemorhous mexicanus), the bacterial pathogen Mycoplasma gallisepticum (MG) causes conjunctivitis, with ocular inflammation mediated by pro- and antiinflammatory cytokines and infection triggering MG-specific antibodies. Here, we tested the role of gut bacteria in host responses to MG by using oral antibiotics to perturb bacteria in the gut of captive house finches prior to experimental inoculation with MG. We found no clear support for an impact of gut bacterial disruption on conjunctival pathology, MG load, or plasma antibody levels. However, there was a non-significant trend for birds with intact gut communities to have greater conjunctival pathology, suggesting a possible impact of gut bacteria on pro-inflammatory cytokine stimulation. Using 16S bacterial rRNA amplicon sequencing, we found dramatic differences in cloacal bacterial community composition between captive, wild-caught house finches in our experiment and free-living finches from the same population, with lower bacterial richness and core communities composed of fewer genera in captive finches. We hypothesize that captivity may have affected the strength of results in this experiment, necessitating further study with this consideration. The abundance of anthropogenic impacts on wildlife and their bacterial communities, alongside the emergence and spread of infectious diseases, highlights the importance of studies addressing the role of commensal bacteria in health and disease, and the consequences of gut bacterial shifts on wild hosts.
- An Assessment of the Relationship among Oxidative Stress, Adaptive Immunity and Genetic Variations in the Chicken, Gallus gallusDeng, Hui (Virginia Tech, 2010-09-01)Oxidative stress (OS) has been associated with aging and age-related diseases in humans, as well as with the decline in economic trait performance in poultry and other domesticated animals. However, the potential effects of OS on the poultry immune system are not well understood. In addition, the impact of bird genetic variation on redox balance remains to be elucidated. Thus, the central hypothesis of this dissertation is: The bird's adaptive immunocompetence is impacted by their OS level, which is not only influenced by environmental factors, but also related to genetic phenotype of either mitochondrial DNA (mtDNA) or nuclear DNA (nDNA). In the first phase of this study, White Leghorn chickens were provided ethanol at different concentrations in drinking water to induce OS. Biomarkers including malondialdehyde (MDA), glutathione (GSH), and plasma uric acid (PUA) were measured to assess OS before and after ethanol treatment. The adaptive immune response during an OS event was measured by plasma IgG and IgM levels, major lymphoid organ weights, CD4+/CD8+ cell ratio, and histopathological analysis of the immune organs. Results showed that when OS was induced by 10% ethanol, chicken adaptive immune responses decreased; however, when birds were exposed to 2% ethanol, there was an enhancement in antioxidant defense and immune response; These results would suggest a negative correlation between OS level and chicken adaptive immune response. In the second phase of the study, subsets of chickens were selected based on their high (H)- or low (L)-OS to assess for variations in their genetic phenotypes. Using MDA levels, 36 chickens were chosen to scan a 2734-bp region of mtDNA, but no definitive SNP was detected. In another experiment, 40 chickens were conversely selected according to three biomarkers for OS. Although no variation was found at eight SNP loci tested across the mitochondrial genome, mtDNA damage measured by 8-hydroxy-2′-deoxy-guanosine was shown to increase with time, and at higher levels in the high OS birds (p < 0.05). Thses results suggest that long-term high OS levels in chickens may increase the somatic mutation of their mtDNA. In the final phase of this dissertation, the effect of nDNA on OS, measured via a genome-wide association study was performed with 18 H and 18 L chickens using the latest chicken 60k SNP microarray for genotyping. Among 56,483 SNPs successfully genotyped, 13 SNPs across five independent loci were associated with OS at significance level of p ≤ 0.001, and another 144 SNPs were also associated with OS (p ≤ 0.01). These results indicate new loci and related genes for their genetic influence upon redox balance. In general, experiments carried out on White Leghorn chickens here have shown that adaptive immune response is tightly related to changes of OS. Further, genetic variance in nDNA is associated with the risk of high OS or the ability to better resist it.
- Bacterial profiles and ex vivo effects of Salmonella Heidelberg on leukocyte function in turkey purebred linesPotter, Tiffany Dawn (Virginia Tech, 2014-11-05)Escalating product recalls as a consequence of Salmonella-contaminated poultry products have resulted in detrimental economic impacts. One long-term alternative to Salmonella prevention, receiving increased attention, is selection to improve genetic resistance. This study evaluated the effects of an oral Salmonella Heidelberg (SH) challenge on bacterial colonization, and the ex vivo effects of SH on phagocytic and bactericidal leukocyte function in turkeys from six pedigree lines (A-F). Data were analyzed using JMP Pro (SAS) and differences were determined using Student’s t-test following ANOVA with significance reported at P ≤ 0.05. Interaction effects of treatment X gender X genetic line were significant on bacterial colonization in the ceca. Cumulatively, females exhibited higher phagocytosis potential than males. The main effect of genetic line was significant bactericidal activity of PBMCs. Microbial profiling of cecal DNA was performed to examine differences in colonization of Salmonella, E. coli, and Enterococcus species among the genetic lines. Results indicated line E having the highest Enterococcus but lowest Salmonella colonization than all other lines, while line A birds displayed the highest Salmonella colonization. These results suggest that gender and genetic line have a marked effect on susceptibility to Salmonella colonization, while genetic line X gender has a more eminent effect on Enterococcus cecal colonization. If able to determine genetic markers associated with these immune responses to Salmonella, genetic selection for increased resistance could be feasible in turkeys.
- Cellular Events During Coccidial Infection in ChickensSu, Shengchen (Virginia Tech, 2016-09-21)Avian coccidiosis is caused by the intestinal protozoa Eimeria. The parasite's site of infection in the intestine is site specific. Eimeria acervulina mainly affects the duodenum, E. maxima the jejunum, and E. tenella the ceca. Lesions in the intestinal mucosa cause reduced feed efficiency and body weight gain in Eimeria-challenged chickens. My previous studies showed that the growth reduction may be due to changes in expression of digestive enzymes and nutrient transporters in the intestine. This can also lead to diminished intracellular pools of nutrients and inhibit pathogen replication. In this dissertation, further analysis of cellular events was performed. Expression of host defense peptides (HDPs), apoptosis and autophagy related genes were examined in Eimeria challenged broilers. The results showed that upon Eimeria infection, LEAP2 was consistently downregulated in the target tissues, while the avian beta-defensins (AvBDs) showed many variations in expression patterns. Downregulation of LEAP2 may be a mechanism for Eimeria to combat the host defense system, and to promote its survival inside the host cell. The in situ hybridization results showed that LEAP2 was expressed only along the villus in the small intestine and not in the crypt. This is the first time LEAP2 has been localized to epithelial cells of the chicken intestine. Eimeria infection can also induce an anti-apoptotic and anti-autophagy state in the host cells. This condition can be both favorable and unfavorable to parasite survival and replication inside the host cell. A comparison of gene expression between Ross and Eimeria resistant Fayoumi (line M5.1 and M15.2) chickens challenged with Eimeria maxima was conducted. The comparison among different lines of chickens showed differential gene expression patterns in lines with different resistance to Eimeria. The similar body weight reduction indicated that there may not be a significant Eimeria resistant line among the Ross, Fayoumi M5.1 and M15.2 birds. The interaction between Eimeria and the host cell is very complex. Studying the mechanisms behind the changes of gene expression during Eimeria infection may give rise to potential therapeutic targets of coccidiosis.
- Characterizing physiological and genetic differences in the early immune response to Haemonchus contortus in resistant and susceptible sheepBowdridge, Scott Alexander (Virginia Tech, 2009-10-12)This dissertation compares immune responses of resistant and susceptible sheep to infection with Haemonchus contortus during the peri-parturient period and larval stage of infection. Identification of immunological events resulting in parasite resistance in St. Croix hair sheep may provide better targets for differential gene expression analysis and eventual discovery of selectable markers for parasite resistance. Antibody levels of hair ewes and composite Dorset x Finnsheep-Rambouillet wool ewes were measured during breeding and again after parturition. Results demonstrated that hair ewes had higher levels immunoglobulin-A after infection and maintained a higher level of circulating antigen-specific antibody when compared to wool ewes. To characterize immune responses to the larval stage of infection, hair and wool lambs were sacrificed at 0, 3, 5, and 7 d after infection with H. contortus. Neutrophil migration to abomasal mucosa and lymph node development were higher in hair sheep than in wool sheep. Gene expression analysis indicated no difference in the abomasal lymph node as both breeds expressed a general T-helper cell type 2 (TH2) response. However, profound differences in TH2 responses were observed in the abomasal mucosa, where hair sheep expressed more IL-4, -13 and -33 than wool sheep. These data thus document the presence of immunological differences between the breeds. Immune responses to larval parasite infection in wool sheep are generally suppressed and may increase the magnitude and duration of infection whereas immune responses to larval infection in hair sheep was more robust and more strongly polarized towards a TH2
- Combination of probiotics and coccidiosis vaccine enhances protection against an Eimeria challengeRitzi, Miranda M.; Abdelrahman, Wael; van-Heerden, Kobus; Mohnl, Michaela; Barrett, Nathaniel W.; Dalloul, Rami A. (BMC, 2016-11-08)Coccidiosis is endemic in the commercial broiler industry capable of inflicting devastating economic losses to poultry operations. Vaccines are relatively effective in controlling the disease; their efficacy could potentially be improved with concurrent use of probiotics as evaluated in this study using an Eimeria challenge. Day of hatch 400 Cobb-500 male broilers were assigned to one of four treatment groups including control (CON), vaccine-only gel application (VNC), probiotic-only gel application (NPC), and vaccine-plus-probiotic gel application (VPC). Birds were placed in floor pens (6 replicate pens/treatment, 16–17 birds/pen). NPC and VPC birds received the probiotics in the water on days 2–4, 8, 14–20, 22, 29, and 34–36. On day 15, birds were mildly challenged with 0.5 mL of a mixed oral inoculum of Eimeria sp. prepared with the coccidiosis vaccine at 10× the vaccination dose. Performance measurements were recorded on first day and weekly afterwards, and lesion scores were evaluated 6 days post-challenge. Overall, the probiotics and coccidiosis vaccine resulted in an enhanced protective effect against the challenge, with VPC birds exhibiting lower lesion scores in the duodenum than VNC or NPC birds. Birds in the VPC treatment also demonstrated higher weight gains during days 1–15, days 7–15, and days 21–28 when compared to the VNC birds. These results suggest that the combination of probiotics and coccidiosis vaccines could enhance performance and provide an additional protective effect against a mixed Eimeria challenge.
- Delayed access to feed affects broiler small intestinal morphology and intestinal cell ontogenyLiu, Kuan-Ling (Virginia Tech, 2019-08-01)In the broiler industry, chicks are often deprived of feed and water up to 48 h posthatch. This delayed access to feed (DAF) has been found to inhibit small intestinal development, compromising growth of the chick. To further understand the impact of DAF on small intestines at the molecular level, many developmental genes that regulate intestinal development were investigated. The objective of this study was to determine the effect of DAF on early posthatch broiler small intestinal morphology, which includes villus height (VH) and crypt depth (CD), and to quantify changes in regulatory genes, such as Olfactomedin 4 (Olfm4), Marker of Ki-67 (Ki-67), Peptide Transporter 1 (PepT1), and Mucin 2 (Muc2), in response to DAF. The Olfm4 mRNA can clearly identify stem cells in the intestinal crypt, which allows VH and CD to be measured, while Ki-67 marks the proliferating cells. The peptide transporter PepT1 is located on intestinal epithelial cells and plays a critical role in transporting di- and tripeptides. Muc2, which is secreted from goblet cells, forms mucus that lines the intestinal epithelial cells acting as a layer of protective coating. Cobb 500 chicks, hatching within a 12 h window, were randomly allocated into three experimental groups: control with no feed delay (ND), 24 h feed delay (D24), and 36 h feed delay (D36). Quantification of Olfm4, Ki-67, PepT1, and Muc2 mRNA abundance were investigated by quantitative PCR, in duodenum, jejunum, and ileum at 0 h, 24 h, 36 h, 72 h, 120 h, and 168 h posthatch. Additionally, localization of cells expressing each gene was visualized using in-situ hybridization at all listed times except 168 h posthatch. Statistical analysis was performed using JMP Pro 14, and significant differences between treatments within a collection day were determined by t-test and one-way ANOVA (P < 0.05). In the ND group, duodenal CD at 0 h was greatest compared to all other time points. With DAF, the duodenal VH of D36 chicks was lower at 36 h (P < 0.001) and 72 h (P = 0.002) compared to ND chicks. In the jejunum and ileum, the VH of D36 chicks was lower at 120 h (P = 0.005) and 72 h (P = 0.03), respectively, compared to ND chicks. In contrast, the VH of D24 chicks at 24 h was greater than ND (P = 0.004) in the jejunum. There was no difference between treatments by 168 h in all intestinal segments. The CD was also lower in DAF groups compared to ND but only in the jejunum and ileum. In contrast, duodenal CD was greater in D24 chicks at 24 h (P = 0.039) and in D36 chicks at 36 h (P < 0.0001) compared to ND chicks, but the difference was no longer significant by 72 h. The VH/CD ratio was lower in all three segments, except the ileum displayed a greater VH/CD ratio in D24 and D36 chicks at 24 h and 36 h, respectively, compared to ND chicks. The mRNA abundance of Olfm4 and Ki-67 was greater in DAF groups upon refeeding, but not until 120 h. The PepT1 mRNA abundance was greater in DAF groups while the abundance of Muc2 mRNA was lower. This difference in mRNA abundance level was more prominent in the duodenum and jejunum. From the analysis of number and distribution of goblet cells found in the upper half and lower half of the villi, expressed as a ratio (VU/VL), a greater ratio was observed in delayed groups compared to ND. In summary, while DAF resulted in altered small intestinal morphology with an effect more pronounced in D36 than D24 chicks, upon refeeding, some genes important to intestinal development were upregulated as a response to the treatment.
- Development and validation of a house finch interleukin-1β (HfIL-1β) ELISA systemKim, Sungwon; Park, Myeongseon; Leon, Ariel E.; Adelman, James S.; Hawley, Dana M.; Dalloul, Rami A. (2017-08-30)Background A unique clade of the bacterium Mycoplasma gallisepticum (MG), which causes chronic respiratory disease in poultry, has resulted in annual epidemics of conjunctivitis in North American house finches since the 1990s. Currently, few immunological tools have been validated for this songbird species. Interleukin-1β (IL-1β) is a prototypic multifunctional cytokine and can affect almost every cell type during Mycoplasma infection. The overall goal of this study was to develop and validate a direct ELISA assay for house finch IL-1β (HfIL-1β) using a cross-reactive chicken antibody. Methods A direct ELISA approach was used to develop this system using two different coating methods, carbonate and dehydration. In both methods, antigens (recombinant HfIL-1b or house finch plasma) were serially diluted in carbonate-bicarbonate coating buffer and either incubated at 4 °C overnight or at 60 °C on a heating block for 2 hr. To generate the standard curve, rHfIL-1b protein was serially diluted at 0, 3, 6, 9, 12, 15, 18, 21, and 24 ng/mL. Following blocking and washing, anti-chicken IL-1b polyclonal antibody was added, plates were later incubated with detecting antibodies, and reactions developed with tetramethylbenzidine solution. Results A commercially available anti-chicken IL-1β (ChIL-1β) polyclonal antibody (pAb) cross-reacted with house finch plasma IL-1β as well as bacterially expressed recombinant house finch IL-1β (rHfIL-1β) in immunoblotting assays. In a direct ELISA system, rHfIL-1β could not be detected by an anti-ChIL-1β pAb when the antigen was coated with carbonate-bicarbonate buffer at 4°C overnight. However, rHfIL-1β was detected by the anti-ChIL-1β pAb when the antigen was coated using a dehydration method by heat (60°C). Using the developed direct ELISA for HfIL-1β with commercial anti-ChIL-1β pAb, we were able to measure plasma IL-1β levels from house finches. Conclusions Based on high amino acid sequence homology, we hypothesized and demonstrated cross-reactivity of anti-ChIL-1β pAb and HfIL-1β. Then, we developed and validated a direct ELISA system for HfIL-1β using a commercial anti-ChIL-1β pAb by measuring plasma HfIL-1β in house finches.
- Developmental gene expression of host defense peptides in immune organs and the small intestine of turkey poults (Meleagris gallopavo)Hamad, Shaimaa Kamal (Virginia Tech, 2016-09-28)Host defense peptides (HDPs) are a large group of small positively charged peptides that play an important role in innate immunity. Their role is more critical at early ages when other components of the immune system have not fully developed. There are three classes of avian HDPs: avian beta defensins (AvBDs), cathelicidins (Cath) and liver-expressed antimicrobial peptide 2 (LEAP-2). The objective was to compare expression of HDPs in male turkey poults at day of hatch (D0), D7, D14, D21 and D28 from the thymus, spleen, bursa, duodenum, jejunum and ileum. The expression of AvBD1, AvBD2, AvBD8, AvBD9, AvBD10, AvBD13, Cath2, Cath3 and LEAP-2 was measured using qPCR (n=6 birds/tissue/age). Data were analyzed by one-way ANOVA and Tukey's test, and significance considered at P ≤ 0.05. AvBDs and Caths exhibited greater expression in immune organs than intestinal tissues, with the greatest expression of AvBDs observed in the spleen. The intestinal tissues showed very low expression of AvBDs except for AvBD10 at D0. Similar to AvBDs, Caths expression in the immune organs was greater than the intestinal tissues with the spleen having the greatest expression among immune organs. Conversely, LEAP-2 showed greater expression in the intestinal tissues than in the immune tissues, which showed very low LEAP-2 expression unlike other HDPs. Understanding the differential expression of HDPs could reveal the innate immune status of poults, and may subsequently allow improvement of their health through appropriate mitigation strategies.
- Dietary Non-Drug Feed Additive as an Alternative for Antibiotic Growth Promoters for Broilers During a Necrotic Enteritis ChallengeCalik, Ali; Omara, Islam I.; White, Mallory B.; Evans, Nicholas P.; Karnezos, T. Peter; Dalloul, Rami A. (MDPI, 2019-08-13)Necrotic enteritis, caused by Clostridium perfringens, is an enteric disease that leads to poor performance and increased mortality, resulting in significant economic losses in poultry production. This study evaluated the effects of a proprietary prebiotic, probiotic, and plant extract blend on performance of broilers during coccidiosis challenge leading to necrotic enteritis (NE). In total, 744 Cobb500 male broilers were randomly allocated to 3 treatments (8 replicates, 31 birds/pen) including, the negative control (NC) fed a basal diet; the positive control (PC) fed a basal diet with Virginiamycin; and the additive group fed basal diet with a blend of prebiotic, probiotic, and plant extract (BSN). A unique, naturally occurring NE model developed to mimic field conditions was implemented to challenge the birds. This model consists of spraying a concentrated commercial coccidiosis vaccine on litter and feed upon bird placement, which, in conjunction with the presence of C. perfringens spores in the environment, leads to the development of a NE outbreak one week post vaccine application. At the onset of NE on d7, three birds/pen were selected for scoring NE lesions. Body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) were recorded on days 7, 14, 28, and 42. Carcass composition was assessed by dual energy X-ray absorptiometry (DXA) analysis on day 42. Dietary supplementation of BSN significantly (p < 0.05) improved FCR during starter and grower periods. Dietary treatments had no effect on NE lesions in the small intestine. DXA analysis revealed slightly higher lean content in BSN birds compared to NC. These results showed that dietary supplementation of the BSN blend significantly improved broilers performance during the early NE challenge phase, as well as in the grower period.
- Differential appetite regulation in lines of chickens selected for high and low juvenile body weight: the role of beta-MSHSmith, Marissa L. (Virginia Tech, 2011-03-18)Melanocortins play a key role in appetite regulation across species. One such melanocortin, beta-melanocyte stimulating hormone (beta-MSH) is receiving increasing attention for its anorexigenic effects. In chicks selected for low (LWS) and high (HWS) juvenile body weight, beta-MSH differentially decreased food intake and HWS chicks may be more sensitive to its effects. Both lines responded similarly to beta-MSH with decreased water intake. While whole blood glucose concentrations and ingestive and non-ingestive behaviors (sit, stand, preen, perch, deep rest, jumps, escape attempts, feed pecks, defecations, and total distance traveled) were not affected in either line, beta-MSH increased corticosterone in LWS chicks but not HWS chicks. However, despite the increase in corticosterone concentration in LWS, astressin, a corticotrophin releasing hormone (CRH) receptor antagonist, did not attenuate the effects of beta-MSH in either line suggesting that the altered stress response may not be acting via CRH receptors. When beta-MSH was co-administered with HS014, a highly selective antagonist for the melanocortin 4 receptor, only LWS responded with an attenuated response to beta-MSH suggesting that the differential response may in part be due to altered receptor affinity or binding resulting from the selection process. To investigate the roles of the hypothalamus and hindbrain in the differential food intake response, an experiment was designed where chicks were injected targeting either the lateral or 4th ventricle utilizing a novel freehand injection procedure. Chicks from both lines responded similarly to beta-MSH following both lateral and 4th ventricle injections. Together, these data suggest that alterations in the b-melanocortinergic appetite regulation system may be in part responsible for the differential body weights of the LWS and HWS lines. [Adaptations of chapters II, III, and IV have been published in Neuroscience Letters, Journal of Neuroendocrinology, and Behavioural Brain Research, respectively]
- Differing house Finch cytokine expression responses to Original and evolved isolates of Mycoplasma gallisepticumVinkler, Michal; Leon, Ariel E.; Kirkpatrick, Laila T.; Dalloul, Rami A.; Hawley, Dana M. (Frontiers, 2018-01-22)The recent emergence of the poultry bacterial pathogen Mycoplasma gallisepticum (MG) in free-living house finches (Haemorhous mexicanus), which causes mycoplasmal conjunctivitis in this passerine bird species, resulted in a rapid coevolutionary arms-race between MG and its novel avian host. Despite extensive research on the ecological and evolutionary dynamics of this host–pathogen system over the past two decades, the immunological responses of house finches to MG infection remain poorly understood. We developed seven new probe-based one-step quantitative reverse transcription polymerase chain reaction assays to investigate mRNA expression of house finch cytokine genes (IL1B, IL6, IL10, IL18, TGFB2, TNFSF15, and CXCLi2, syn. IL8L). These assays were then used to describe cytokine transcription profiles in a panel of 15 house finch tissues collected at three distinct time points during MG infection. Based on initial screening that indicated strong pro-inflammatory cytokine expression during MG infection at the periorbital sites in particular, we selected two key house finch tissues for further characterization: the nictitating membrane, i.e., the internal eyelid in direct contact with MG, and the Harderian gland, the secondary lymphoid tissue responsible for regulation of periorbital immunity. We characterized cytokine responses in these two tissues for 60 house finches experimentally inoculated either with media alone (sham) or one of two MG isolates: the earliest known pathogen isolate from house finches (VA1994) or an evolutionarily more derived isolate collected in 2006 (NC2006), which is known to be more virulent. We show that the more derived and virulent isolate NC2006, relative to VA1994, triggers stronger local inflammatory cytokine signaling, with peak cytokine expression generally occurring 3–6 days following MG inoculation. We also found that the extent of pro-inflammatory interleukin 1 beta signaling was correlated with conjunctival MG loads and the extent of clinical signs of conjunctivitis, the main pathological effect of MG in house finches. These results suggest that the pathogenicity caused by MG infection in house finches is largely mediated by host pro-inflammatory immune responses, with important implications for the dynamics of host–pathogen coevolution.
- Dissecting the impact of macrophage migration inhibitory factor (MIF) on host immune responsePark, Myeongseon (Virginia Tech, 2018-10-16)Macrophage migration inhibitory factor (MIF) has been implicated in mediating both innate and adaptive immune responses in inflammatory and infectious diseases. The sequence and structure of MIF is highly conserved across the avian phylogeny, which underlies high sequence homology and functional similarities between turkey and chicken MIFs. Turkey MIF (TkMIF) inhibited cell migration and promoted cell proliferation with production of inflammatory mediators, comparable to the biological properties of chicken MIF (ChMIF), thus indicating the biological cross-reactivity between turkey and chicken MIFs. This study identified the cell surface receptor(s) that could bind ChMIF and the biological roles triggered by such interactions. In addition to CD74, a previously identified receptor, CXCR4 also interacts with ChMIF. Moreover, the formation of receptor complexes was shown between CXCR4 and CD74. MIF signaling through CXCR4 and CD74 led to cell chemotaxis and proliferation activity as well as intracellular calcium influx. Intriguingly, Eimeria MIF (EMIF), a homologue secreted following parasitic infection, also interacted with CD74 leading to comparable biological functions to those of ChMIF. Given such observations, we hypothesized that CXCR4 and CD74 are receptors for ChMIF leading to the functional consequences similarly manifested by EMIF interaction with the corresponding receptors. EMIF, predominantly secreted from the invasive merozoite stage, may help the parasite exploit the host immune response by interacting with common ChMIF receptors. This may lead to functional mimicry thus provoking the question of whether EMIF would modulate the biological functions of ChMIF to manipulate the host defense that allows more efficient invasion of the host. To evaluate this concept, a transgenic E. tenella lacking MIF was generated by in vivo passage of E. tenella transfected with a CRISPR plasmid targeting EMIF. Although not fully disrupted, reduction of EMIF expression was observed in the transgenic E. tenella itself as well as in inoculated cells, which resulted in enhanced survival of host cells. Herein, we achieved a better characterization of the functional roles of both avian and parasite MIFs underlying the interaction with common host receptors, along with the essential role of parasite MIF promoting host cell death during parasitic infection.
- The Effect of Dietary Phytic Acid Concentration and Phytase Supplementation on Performance, Bone Ash, and Intestinal Health of Broilers Vaccinated With a Live Coccidial Oocyst VaccineLehman, Regina N. (Virginia Tech, 2011-11-03)The role of nutrition in providing optimal broiler growth and intestinal health is essential, especially during stress or disease challenge. Feed enzymes are useful for improving performance of poultry, particularly when nutrition, management, or health status is not favorable. The objective of the following experiments was to evaluate the effect of dietary phytic acid (PA) and phytase on the performance and intestinal health of birds that were vaccinated with a live coccidial oocyst vaccine. For each experiment, half of the chicks were spray-vaccinated at day-of-hatch with Coccivac®-B and grown out in floor pens with ad libitum access to diets formulated to meet Cobb nutrient recommendations. In the first experiment, birds were given one of three diets that included different levels of a PA solution to obtain dietary PA levels of 0.74, 0.87, and 1.12% for low, medium, and high PA diets, respectively. In the second experiment, two levels of PA were included to obtain dietary PA levels of 0.75 and 1.05% for low and high PA diets, respectively. In addition, phytase was added over the top to half of the diets at 1000 FTU/kg, resulting in four diets: low PA without phytase, low PA with phytase, high PA without phytase, and high PA with phytase. Live performance parameters including body weight, body weight gain, feed intake, feed conversion, and mortality were measured as well as tibia ash (experiment 2) and indicators of small intestinal health including morphology, apparent ileal amino acid digestibility (IAAD), and pH (experiment 2). The results presented here indicated that giving broilers vaccinated against coccidiosis a medium level of PA was detrimental to feed intake, body weight gain, and it induced necrotic enteritis (P ≤ 0.05). Adding phytase on top of nutritionally adequate diets did not improve performance (P ≥ 0.05), but did improve (P ≤ 0.05) apparent IAAD and morphology of the small intestine, especially in younger birds. In addition, it has been determined that important considerations in diet formulation also can include the phytate: protein as well as calcium: total phosphorus ratios, as these may critically affect how phytate impacts bird health and performance.
- Effect of probiotics or high incubation temperature on gene expression and cell organization of the small intestine and yolk sac of chicksJia, Meiting (Virginia Tech, 2021-11-30)The small intestine and yolk sac (YS) are important organs for nutrient absorption and innate immunity in chickens during the post-hatch or prehatch periods. These organs share a similar structure of epithelial cell-lined villi with tight junctions between adjacent cells. Probiotics have been reported to improve chicken growth performance and gut health including promotion of intestinal morphology. However, there are few studies that show the effect of probiotics on ontogeny of intestinal epithelial cells and antimicrobial peptides, or intestinal integrity in young healthy chicks. Heat stress during incubation was shown to increase mortality and decrease hatchability of chicks, while no studies have investigated the effect of heat stress on the integrity of the YS, which might be related to hatching performance. There were four studies conducted in this research: 1) a comparison of the effect of two probiotics on the ontogeny of small intestinal epithelial cells in young chicks; 2) the effect of two probiotics on mRNA abundance of tight junction proteins in the small intestine of young chicks; 3) the effect of high incubation temperature on mRNA abundance of tight junction proteins in the YS of broiler embryos; and 4) comparison of avian defense peptide mRNA abundance in the YS of broilers and layers. In study 1, Probiotics transiently decreased body weight gain (BWG) from day 2 to day 4, but did not affect body weight (BW) from day 2 to day 8, and small intestinal weight and intestinal morphology from day 2 to day 6. Probiotics did not affect marker gene expression of intestinal stem cells (Olfm4) and goblet cells (Muc2) in all small intestinal segments, but did increase expression of a marker gene of proliferating cells (Ki67), and decreased an antimicrobial peptide (liver-enriched antimicrobial peptide 2, LEAP2) in the jejunum at day 4. Probiotic 1 decreased PepT1, a marker of enterocytes in the duodenum at day 4. These results suggest that probiotics did not improve growth performance and intestinal morphology in young healthy chicks, but temporarily promoted intestinal epithelial cell proliferation and decreased LEAP2 antimicrobial peptide expression in the jejunum. In situ hybridization (ISH) showed that Ki67+ proliferating cells were mainly located in the crypt region and the blood vessels of villi. In study 2, Probiotic supplementation to newly hatched chicks for less than one week did not affect mRNA abundance of the tight junction proteins in the small intestine. Occludin (OCLN) mRNA, which was detected by ISH to be expressed in intestinal epithelial cells in both the villus and crypt regions, was greater in the duodenum of female chicks than males. In study 3, high incubation temperature starting from embryonic day 12 (E12) affected mRNA abundance of the tight junction proteins in the YS, including increased zonula occluden 1 (ZO1) at E13, increased junctional adhesion molecule A (JAMA) and heat shock protein 90 (HSP90) at E17, but decreased tight junction protein JAMA at E19 and OCLN at day of hatch (DOH). These results showed that the YS tight junction proteins were increased by short term heat exposure but decreased by long term heat exposure. In study 4, the expression of avian β defensin 10 (AvBD10), CATHs and toll-like receptors in the YS was examined. Toll-like receptors were highly expressed in the YS at early incubation stages (E7), while CATHs showed a peak expression from E9 to E13, which was similar to the expression pattern of AvBD10. CATHs and AvBD10 mRNA temporal expression patterns were similar in broilers and layers, while their expression levels were different. Layers, especially brown layers, had greater mRNA abundance for antimicrobial peptides such as AvBD10, CATH1, and CATH2 in the YS. These results demonstrate that the antimicrobial peptide temporal expression patterns in the YS are not affected by breed, but their expression levels are affected by breed. In summary, the small intestine and the YS are essential for nutrient uptake, innate immunity, and maintenance of integrity. The ontogeny of intestinal epithelial cells, such as proliferating cells can be modulated by probiotic supplementation. Similar to the small intestine, the YS can also express tight junction proteins, which can be affected by high incubation temperature. Antimicrobial peptide expression in the intestine of healthy young chicks is also transiently decreased by probiotic supplements. Avian defensin and cathelicidin expression patterns in the YS were not affected by breed.
- Effects of a dietary antioxidant blend on growth performance, liver function, oxidative stress, and meat and fat quality in pigs and broiler chickens fed diets high in oxidantsLu, Ting (Virginia Tech, 2013-08-22)High feed ingredient prices have increased the use of by-products containing a high proportion of polyunsaturated fatty acids (PUFA) in pig and chicken feeds. This can increase the oxidation of other feed nutrients as well as causing oxidative stress in animals. Two studies were conducted to evaluate the effects of a dietary antioxidant blend (AOX, ethoxyquin and propyl gallate) in pigs and broiler chickens fed a diet high in oxidants. The objective of the first study was to evaluate the antioxidant blend on growth performance, meat quality, liver function, oxidative status, carcass characteristics, meat quality, and fatty acid profile in pigs. Crossbred barrows (n = 100, 10.91 ± 0.65 kg, 36 ± 2 d of age, Landrace × Duroc) were allotted to 5 treatments based on body weight (BW, 5 replicate pens per treatment, 4 pigs per pen). Treatments included: 1) HO: high oxidant diet containing 5% oxidized soy oil and 10% PUFA source (containing docosahexaenoic acid, DHA, 3.7% of diet); 2) VE: the HO diet with 11 IU/kg of added vitamin E; 3) AOX: the HO diet with AOX (135 mg/kg); 4) VE+AOX: the HO diet with both vitamin E and AOX; and 5) SC: a standard corn-soy control diet. The trial lasted for 118 d; on d 83, the HO diet pigs were switched to the SC diet because the animals were displaying very poor health. Compared with SC pigs, HO pigs had decreased average daily gain (ADG) and average daily feed intake (ADFI) from d 26 to 82 (P < 0.05). However, after switching the HO pigs to the SC diet, the VE treatment became the most stressed treatment with the poorest performance from d 83 to 118 (P < 0.05). The AOX restored pig performance to a level similar to pigs fed the SC diet (P > 0.05) with greater gain to feed ratio (G:F) for the entire period (P < 0.05). The AOX added treatments also attenuated the enlarged liver symptoms and reduced markers of liver stress including total bilirubin and aspartate transaminase, thiobarbituric acid reactive substances (TBARS) and carbonyl concentrations. In addition, the AOX addition in the high oxidant diet restored the lighter carcass weight, less back fat, less lean body mass and smaller loin eye area, decreased dressing percentage and intensive lipofuscin deposition induced by the high oxidant diet. However, the traits of loin muscle redness and belly firmness were not fully corrected by AOX. The second study was to investigate the antioxidant blend and vitamin E on growth performance, oxidative status, meat quality, fatty acid profile, liver function and inflammatory response in broiler chickens. Cobb 500 male broilers (n = 1200, 44.7 ± 0.8 g, d 0) were randomly distributed into 60 floor pens across 6 treatments with 10 replicate pens of 20 chicks each. Treatments included: 1) HO: high oxidant diet with vitamin E at 10 IU/kg, 3% oxidized oil, 3% PUFA source; 2) VE: the HO diet with vitamin E at 200 IU/kg; 3) AOX: the HO diet with AOX at 135 mg/kg, 4) VE+AOX: the HO diet with both vitamin E at 200 IU/kg and AOX at 135 mg/kg, 5) SC: standard control, a corn soy diet with vitamin E at 10 IU/kg, 3% non-oxidized soybean oil, no PUFA source, and 6) PC: positive control, the SC diet with AOX at 135 mg/kg. Compared to the SC birds, the PUFA added treatments (HO, VE, AOX, VE+AOX) groups had greater body weight, ADG and ADFI from d 0 through d 21 (P < 0.05). However, the growth of birds fed the VE treatment fell behind that of other treatments (P < 0.05) during the last 21 d of the trial. Compared to the HO birds, the AOX birds had lower TBARS and greater uric acid concentrations in the plasma, greater gene expression of superoxide dismutase and less drip loss, suggesting enhanced systematic antioxidant capability. In addition, dietary addition of AOX or AOX plus VE moderately improved liver function and reduced inflammation in fat tissue to a level similar to control groups. In both studies, the AOX supplement was effective in preserving PUFA, especially DHA deposition in the back fat of pigs and abdominal fat of chickens. These results suggest that feeding the high oxidant diet caused a series of changes in growth performance, liver function, oxidative status, carcass characteristics and meat quality in pigs, and AOX addition attenuated many of these. The supplementation of AOX also showed some effects on reducing oxidative stress in chickens. However, the effects were not as profound as the pig study.
- Effects of Calcium and Enzyme Supplementation on the Occurrence of Necrotic EnteritisPaiva, Diego Moreira (Virginia Tech, 2013-01-21)Diet composition and nutrient balance can have a critical impact on intestinal integrity during exposure to enteric pathogens. Researchers have extensively reported benefits on nutrient availability and broiler performance as a consequence of the impact of phytase supplementation. However, the poultry industry has little information on the effects of phytase supplementation in disease settings. The objective of these studies was to evaluate phytase supplementation impact on bird performance, intestinal morphology and pH, nutrient digestibility and bone mineralization during necrotic enteritis (NE). In each experiment, Cobb 500 broilers were obtained from a commercial hatchery and housed in floor pens at the Virginia Tech Turkey Research Center. Birds were placed on used litter from a previous flock that had presented clinical signs of NE. Broilers were fed non-medicated diets formulated to meet NRC (1994) nutrient requirements, except for calcium and phosphorus. In the first experiment, birds began exhibiting clinical signs of NE on d 9, and elevated NE-associated mortality persisted until d 26. Mortality was influenced by the main effects of dietary Ca or phytase. Dietary Ca supplemented at 0.9% or 1000 FTU/kg of phytase increased mortality compared to 0.6% Ca or 0 FTU/kg phytase, respectively, from d 0 to 19. Feed intake (FI) and feed conversion (FC) were affected by Ca x P interaction. From d 0 to 19, birds fed 0.9% Ca and 0.3% available P (avP) had decreased FI and improved FC compared to birds fed 0.9% Ca and 0.45% avP, while FI and FC were similar in birds fed diets with 0.6% Ca, regardless of avP level. Calcium x P x phytase interaction influenced BW or BWG from d 0-12. In general, birds fed 0.9% Ca and 0.45% avP with phytase were heavier compared to birds fed 0.6% Ca, 0.45% avP, and phytase. Calcium at 0.9% increased gizzard (d 19) and jejunum (d 12) pH. Dietary Ca supplemented at 0.9%, avP supplemented at 0.45%, and 1,000 FTU/kg phytase significantly increased tibia ash weight compared to 0.6% Ca, 0.3% avP, and 0 FTU/kg phytase, respectively, on d 12. A 3-way interaction was observed on d 35 for tibia ash percentage; birds fed 0.9% Ca and 0.45% avP had a significant increase in tibia ash percentage, regardless of phytase supplementation. A 3-way interaction was also observed for Ca and P digestibility on d 35. Phytase supplementation significantly increased Ca digestibility regardless of Ca and P levels of the diets. In addition, diets containing 0.6% Ca and 1,000 FTU/Kg of phytase resulted in a significant increase in P digestibility, regardless of P levels. In the second experiment, birds also began exhibiting clinical signs of NE on d 9, and elevated NE-associated mortality persisted until the end of the trial (d 21). Mortality was significantly affected by an interaction between Ca source and Ca levels. Significantly higher mortality was observed when animals were fed 0.9% Ca diets formulated with calcified seaweed from d 0-21 compared to 0.6% Ca diets (regardless of Ca source). From d 0-7, birds fed 0.6% Ca in diets supplemented with phytase had heavier BW than the other treatments regardless of Ca source. From d 0-14 and 0-21, animals fed diets with calcified seaweed had significantly higher FC than animals fed diets with limestone. On d 21, the gizzard of birds fed 0.9% Ca was significantly less acidic than the gizzard of birds fed 0.6% Ca. In conclusion, reducing dietary levels of Ca associated with phytase supplementation improved bird performance and nutrient digestibility. In addition, these experiments indicate that Ca is an important dietary factor in the pathogenesis of NE.
- Effects of dietary enzyme supplementation on performance, bone ash, small intestinal morphology, and apparent ileal amino acid digestibility of broilers exposed to a live coccidia oocyst vaccineWalk, Carrie L. (Virginia Tech, 2009-09-03)A series of experiments were conducted to evaluate the effects of dietary enzyme supplementation in broilers exposed to a live coccidia oocyst vaccine. In each experiment, Cobb 500 broilers were obtained at day of hatch from a commercial hatchery. Half of the chicks were sprayed with Coccivac BTM and housed in battery brooders in experiment (EXP) 1 and 2 or floor pens (EXP 3). Experimental diets were formulated according to Cobb 500 nutrient recommendations (positive control; PC) with the exception of Ca and available P (aP), which were reduced in the negative control (NC) diets approximately 0.1% (EXP 1), 0.11 and 0.13%, respectively (EXP 2), and 0.13% (EXP 3). Negative control diets in EXP 1 were supplemented with phytase (PHY), protease (PRO), xylanase (XYL), and the combination of PHY+PRO, PHY+XYL, and PHY+PRO+XYL. Negative control diets in EXP 2 were supplemented with PHY A, PHY B, and PHY C. In EXP 3, PC diets were supplemented with PHY at 1000 FTU/kg, and NC diets were supplemented with PHY at 1000 or 5000 FTU/kg. In all three experiments PHY supplementation generally improved (P ≤ 0.05) broiler performance and bone ash. Vaccination reduced (P ≤ 0.05) broiler performance in EXP 1 and 3, but increased (P ≤ 0.05) broiler performance in EXP 2. Xylanase and/or PHY supplementation tended to improve (P = 0.10) ileal amino acid digestibility (IAAD) in vaccinated broilers in EXP 1 and EXP 3. Phytase supplementation improved (P ≤ 0.05) IAAD and vaccination reduced (P ≤ 0.05) IAAD in EXP 3. Small intestinal morphology and goblet cell numbers were affected by enzyme supplementation and vaccination, which resulted in significant (P ≤ 0.05) interactions. In general, vaccination increased (P ≤ 0.05) small intestinal crypt depth and reduced (P ≤ 0.05) goblet cell numbers in EXP 3. Phytase supplementation of the NC diets fed to vaccinated broilers ameliorated the reduction in ileal goblet cells associated with vaccination (EXP 3). Enzyme supplementation tended to improve nutrient digestibility and altered small intestinal morphology in vaccinated broilers. Vaccination reduced broiler performance but nutrient digestion and dietary enzymes may improve nutrient utilization during a coccidia vaccination.
- The effects of dietary β-glucan supplementation on performance and immune response of broiler chicks during an Eimeria challengeCox, Chasity Marie (Virginia Tech, 2009-12-09)Escalating consumer concerns have placed the poultry industry under mounting pressure to reduce the use of chemotherapeutic agents as feed additives. One possible alternative receiving increased attention is the use of immunomodulators such as β-glucan. A pilot study evaluated the effects of a yeast derived β-glucan (Auxoferm YGT) on growth performance and immune response of broiler chickens. Day-old chicks were fed a diet containing 0, 0.02, or 0.1% yeast β-glucan. On days 7 and 14 post-hatch, body weight and relative immune organ weights were measured, peripheral blood was collected to determine heterophil to lymphocyte (H:L) ratios, and small intestinal sections were sampled to evaluate relative gene expression. The addition of β-glucan had no influence on growth. Dietary β-glucan supplementation modulated the expression of interleukin (IL)-8, IL-18, interferon (IFN)-γ and inducible nitric oxide synthase (iNOS) in the small intestine. A subsequent study was conducted to investigate the effects of dietary β-glucan on broiler chick (1440 birds) performance and immune response during a mixed Eimeria infection (day 8 of age). Measurements were taken and samples collected on days 4, 7, 10, 14 and 21 post-hatch. The results from this study show that β-glucan supplementation did not negatively impact performance. The addition of β-glucan to the diet resulted in reduced gross lesion severity and increased H:L ratios. The gene expression results suggest that β-glucans are capable of skewing the host immune response toward aTh1 mediated response and consequently down-regulating the Th2 mediated response.
- «
- 1 (current)
- 2
- 3
- »