Search

Microorganisms are absolutely critical to myriad aspects of the human existence. As a field of study, microbiology could and should serve a greater role on our campus, as it has key connections with many of the Destination and Strategic Growth Areas. We propose the development of a broad concept area in microbiology that will serve as a nexus, as it is applied to solve critical global challenges related to food, energy, water and health, by bridging across multiple disciplines at Virginia Tech (VT). There is increasing recognition of microbes as a driving force in natural and managed environments, biological processes, and ecological structure. Conversely, the importance of culture and individual behavior in affecting microbial communities has also become apparent....

Cleaning and disinfection are critical areas of Veterinary hospital biosecurity. Without a validated biosecurity program, veterinary hospitals are in a vulnerable state. Potential outbreaks of pathogenic microorganisms could occur causing loss of: patient lives, hospital revenue and hospital prestige. Instances of pathogenic outbreaks have been recorded in small animal settings (i.e. veterinary clinics and shelters) and in large animal settings (i.e. farms and hospitals.) Veterinary hospitals, often considered the gold standard of veterinary care are not immune to biosecurity breaches. In last 5 years 82% of veterinary teaching hospitals reported nosocomial infections (Anderson, 2010). The aim of this project was to validate cleaning and sanitation procedures that are already in place in the Harry T. Peters, Jr. Large Animal Hospital located in Blacksburg, VA. This project compared two sanitation protocols and validated them using environmental sampling. Using chi-square analysis results indicated there was no significant difference between the sanitation methods. MALS

Two experiments were conducted to determine the effects of various alternatives to antibiotics in broilers grown on used litter on performance, intestinal lesion scores, body composition, and cecal volatile fatty acid (VFA) concentrations. The first experiment used a corn-soybean meal based basal diet to generate a negative control (NC) diet (without treatment), a Bacillus licheniformis probiotic containing diet (NC + 0.10% DFM1), and four diets that contained various concentrations of a Bacillus subtilis probiotic (NC + 0.05% DFM2, NC + 0.10% DFM2, NC + 0.20% DFM2, and NC + 0.40% DFM2). Experimental diets were fed to broilers over a 42-day period. Body weight gain (BWG) was depressed from d 0-28 and d 0-42 with supplementation of 0.10% DFM1 and 0.05% DFM2 compared to NC fed birds (P ≤ 0.05), with no differences among the remaining treatments and the NC fed birds (P > 0.05). Feed intake (FI) and mortality corrected feed efficiency (FEm) were not different over the 0-42 day period. No differences in oocyst shedding, lesion scores, body composition or cecal VFA production were observed (P > 0.05). Consistently low lesion scores were indicative of a mild coccidial challenge. The second experiment utilized a corn-soybean meal basal diet to generate experimental treatment diets. The basal diet without additional supplements was fed to two groups of control birds, one on clean pine shaving litter (PC) and a second on used litter (NC). Treatment groups were fed the same diet, supplemented with butyrate for the first 14 days (NC + But 0-14), botanicals from d 15-43 (NC + Bot 15-43), butyrate from d 0-14 and botanicals from d 15-43 (NC + But 1-14/Bot 15-43), and butyrate from d 0-43 (NC + But 1-43). Butyrate supplementation reduced BWG from d 0-14 (P ≤ 0.05), but there were no differences in BWG from 0-43 d (P > 0.05). Feed intake was reduced for the NC + But 0-14 group from d 0-43 in comparison to both PC and NC. Lesion scores in the jejunum were reduced with supplementation of butyrate alone, fed for either d 0-14 or d 0-43 (P ≤ 0.05) and is an indication of a mild coccidial infection due to the used litter. Although there were no significant differences among treatments, overall performance was above industry expectations, likely due to the mild fall weather. These two experiments indicate that there are numerous factors involved in the efficacy of antibiotic alternatives, at least partially explaining the inconsistent results observed in the published literature. Master of Science Recently, increased regulations and customer demand have restricted and reduced the use of antibiotics in the poultry industry. Historically, antibiotics were used in poultry feed at sub-therapeutic levels to improve performance and prevent the incidence of clinical and subclinical disease. Since the reduction of antibiotic use, many producers have experienced reduced performance resulting in reduced profits. The limited use of antibiotics can also present an animal welfare issue associated with increased sub-clinical and clinical disease. Many researchers are investigating alternative feed additives that will both improve performance and prevent disease, including probiotics, organic acids, and botanical products. Previous research has demonstrated the ability of these alternative to positively, if inconsistently, influence the performance and health of broiler chickens. Therefore, the objective of this thesis is to investigate the effects of two probiotic bacteria fed at various concentrations and the effects of butyric acid and botanicals, fed alone, or in combination, on the performance, intestinal lesion scores, body composition, and cecal volatile fatty acid concentrations of broiler chickens raised on used litter. In these experiments, the used litter is representative of a mild disease challenge that would likely be present in a commercial poultry setting. Overall, there were few differences with the treatment of broilers with probiotic bacteria, but the lowest inclusions slightly reduced body weight gain compared to the control fed broilers. Butyric acid supplementation reduced body weight gain over the first 14 days, but these differences were no longer observed over the 0 to 42 day period. Although performance was not improved, butyrate did result in reduced intestinal lesion scores from the middle section of the small intestine, indicating potential health benefits with butyrate treatment. Overall, the data presented in this thesis suggest that there are a variety of factors that can alter the effectiveness of these alternatives in broiler production and care should be used in selection of antibiotic replacement tools.

The prophylactic use of antibiotics is rapidly being phased out of poultry operations. Diseases such as coccidiosis are extremely costly, and typically prevented with coccidiostats and antibiotics. New regulation banning antibiotics is mandating a search for new alternatives, such as probiotics and phytogenic feed additives (PFAs). Two separate studies were performed to evaluate the performance and microbial profiles of commercial broilers fed either PFAs or Eimeria challenged broilers fed probiotics. During the probiotics study, 1056 day-old male Ross broiler chicks were assigned to one of 6 treatments, 8 replicates per treatment, with 22 birds per floor pen in a 2x3 factorial arrangement, with DFM mix and coccidiosis challenge being the main factors. Treatments included a non-infected control (NC), a non-infected low level DFM-fed group (DL), a non-infected high level DFM-fed group (DH), an infected control (PC), an infected low level DFM-fed group (DL+), and an infected high level DFM-fed group (DH+). Performance measurements were taken on d0, d7, d14, d21, and d28. Mucosal scrapings from the ileum were obtained on d7, d14, d21, and d28 to determine microbial profiles. On d15, birds in the three infected groups were challenged with a mixed inoculum containing Eimeria maxima, E. tenella, and E. acervulina. Lesion scores were assessed on d21. Body composition was analyzed via Dual Energy X-Ray Absorptiometry (DXA) on d28. Results indicated a trend (P < 0.1) toward improved D28 body composition, lower Clostridium in DFM-fed challenged broilers compared to PC, and higher Bacillus in DH compared to DH+. Results were not significant for BW, BWG, FI, or lesion scores. The second study evaluated the effects of PFA provided as a dietary supplement over a 6-week period on performance and immune responses in broilers. In total, 1056 day-old male chicks were randomly assigned to 8 replicate pens (22 birds/pen) with 6 treatments including a standard corn/wheat basal diet, a basal with antibiotics, and 4 various combinations of PFA (PHY 1, 2, 3, and 4). Performance was assessed on day 7 (D7), 14, 28, and 42. Starter diet was fed D0-14, grower D14-28, and finisher D28-42. Microbial profiling was assessed on D7, 14, and 42, and body composition measured on D42 using Dual Energy X-Ray Absorptiometry (DXA). Results varied by week, but cumulatively, body weight gain (BWG) per bird was higher in PHY1 and 4, statistically similar to AGP and CTRL. With feed conversion ratio (FCR), PHY2 was higher than any other treatment between D0-42, while all other treatments were similar. PCR results were not statistically significant; however, DXA results indicated a higher lean to fat ratio in birds from PHY1-4 when compared to AGP, indicating an overall leaner bird in PHY treatments. The results of these studies suggest that diets supplemented with DFMs or PFAs result in improved broiler performance. Master of Science The prophylactic use of antibiotics is rapidly being phased out of poultry operations. Diseases such as coccidiosis are extremely costly, and typically prevented with coccidiostats and antibiotics. New regulation banning antibiotics is mandating a search for new alternatives, such as probiotics and phytogenic feed additives (PFAs). Two separate studies were performed to evaluate the performance and microbial profiles of commercial broilers fed either PFAs or Eimeria challenged broilers fed probiotics. During the probiotics study, 1056 day-old male Ross broiler chicks were assigned to one of 6 treatments, 8 replicates per treatment, with 22 birds per floor pen in a 2x3 factorial arrangement, with DFM mix and coccidiosis challenge being the main factors. Treatments included a non-infected control (NC), a non-infected low level DFM-fed group (DL), a non-infected high level DFM-fed group (DH), an infected control (PC), an infected low level DFM-fed group (DL+), and an infected high level DFM-fed group (DH+). Performance measurements were taken on d0, d7, d14, d21, and d28. Mucosal scrapings from the ileum were obtained on d7, d14, d21, and d28 to determine microbial profiles. On d15, birds in the three infected groups were challenged with a mixed inoculum containing Eimeria maxima, E. tenella, and E. acervulina. Lesion scores were assessed on d21. Body composition was analyzed via Dual Energy X-Ray Absorptiometry (DXA) on d28. Results indicated a trend (P < 0.1) toward improved D28 body composition, lower Clostridium in DFM-fed challenged broilers compared to PC, and higher Bacillus in DH compared to DH+. Results were not significant for BW, BWG, FI, or lesion scores. The second study evaluated the effects of PFA provided as a dietary supplement over a 6-week period on performance and immune responses in broilers. In total, 1056 day-old male chicks were randomly assigned to 8 replicate pens (22 birds/pen) with 6 treatments including a standard corn/wheat basal diet, a basal with antibiotics, and 4 various combinations of PFA (PHY 1, 2, 3, and 4). Performance was assessed on day 7 (D7), 14, 28, and 42. Starter diet was fed D0-14, grower D14-28, and finisher D28-42. Microbial profiling was assessed on D7, 14, and 42, and body composition measured on D42 using Dual Energy X-Ray Absorptiometry (DXA). Results varied by week, but cumulatively, body weight gain (BWG) per bird was higher in PHY1 and 4, statistically similar to AGP and CTRL. With feed conversion ratio (FCR), PHY2 was higher than any other treatment between D0-42, while all other treatments were similar. PCR results were not statistically significant; however, DXA results indicated a higher lean to fat ratio in birds from PHY1-4 when compared to AGP, indicating an overall leaner bird in PHY treatments. The results of these studies suggest that diets supplemented with DFMs or PFAs result in improved broiler performance.

Coccidiosis is endemic in the commercial broiler industry and inflicts devastating economic losses to poultry operations. Probiotics may provide a potential alternative to the prophylactic use of anticoccidials in commercial production. This study evaluated the effects of probiotic applications (feed and water) on bird performance and resistance to a mixed Eimeria infection in commercial broilers. On day of hatch, 1008 commercial male broilers (Cobb 500) were assigned to one of 6 treatments (8 replicate floor pens; 21 birds/pen), including non-infected negative control (NEG), Eimeria-infected positive control (POS), anticoccidial control (0.01% salinomycin, SAL), intermittent high dose water-applied probiotic (WPI), continuous low dose water-applied probiotic (WPC), and feed-supplemented probiotic (FSP). On d15, all birds except those in NEG were challenged with a mixed inoculum of Eimeria acervulina, E. maxima, and E. tenella. Measurements were taken on d7, 15, 21, 28, 35 and 42. Fecal samples were collected from d20-d24 for oocyst counts, and lesion scores were evaluated on d21. Data were analyzed using the Fit Model platform in JMP Pro 10.0 (SAS). Differences in experimental treatments were tested using Tukey HSD following ANOVA with significance reported at P ° 0.05. Overall, NEG birds outperformed all other groups. For performance, the probiotic groups were comparable to the SAL treated birds, except during the 6 days immediately following the Eimeria species challenge, where the SAL birds exhibited better performance. WPC birds had lower duodenal and jejunal lesion scores, indicating a healthier intestine and enhanced resistance to Eimeria species compared to POS. Birds in the WPI treatment shed fewer oocysts in the feces, though this was not a trend for all of the probiotic treatment groups. The results of this study suggest probiotic supplementation without anticoccidials can enhance performance and help alleviate the negative effects of a mixed Eimeria infection. Master of Science

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. Ph. D.