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This study investigated the microbial colonization and maternal influences on the neonatal calf gut microbiome. Microbiome samples were collected from dams (n = 6) and calves (n = 6) using sterile flocked swabs. The vaginal, oral, and fecal bacterial communities were examined from the dam and the fecal community of calves was examined from birth to 60 d of age. Microbial communities varied by anatomical location and age of the calf. Metagenomic analysis 16s ribosomal DNA revealed ten phyla associated with microbiomes of the dam and the same ten phyla associated with calf feces at various time points: Bacteroidetes, Firmicutes, Proteobacteria, Actinobacteria, Cyanobacteria, Verrucomicrobia, Spirochaetes, Tenericutes, Fibrobacteres, and Lentisphaerae. Overall, the calf meconium and fecal microbiome is influenced by a combination of the maternal vagina, oral, and fecal microbiomes. Further studies will be needed to identify the transference mechanisms of maternal microbes to offspring and the associated host-microbial interactions. / Master of Science in Life Sciences / This study investigated the microbial colonization and maternal influences on the neonatal calf gut microbiome. Microbiome samples were collected from dams (n = 6) and calves (n = 6) using sterile flocked swabs. The vaginal, oral, and fecal bacterial communities were examined from the dam and the fecal community of calves was examined from birth to 60 d of age. Microbial communities varied by anatomical location and age of the calf. Metagenomic analysis 16s ribosomal DNA revealed ten phyla associated with microbiomes of the dam and the same ten phyla associated with calf feces at various time points: Bacteroidetes, Firmicutes, Proteobacteria, Actinobacteria, Cyanobacteria, Verrucomicrobia, Spirochaetes, Tenericutes, Fibrobacteres, and Lentisphaerae. Overall, the calf meconium and fecal microbiome is influenced by a combination of the maternal vagina, oral, and fecal microbiomes. Further studies will be needed to identify the transference mechanisms of maternal microbes to offspring and the associated host-microbial interactions.

In experiment one, week old Jersey bull calves (n=39) were assigned to one of four diets: 21/21 (n=8), 27/33 (n=8), 29/16 (n=9), MILK; or a baseline sacrifice group (n=6). Diets 21/21, 27/33, and 29/16 were milk replacers containing 21, 27, or 29% CP, and 21, 33, and 16% fat, respectively. Diet 21/21 was fed at 15% of BW. Diets 27/33, 29/16, and MILK supplied 180g CP/d. Calves were fed 4 wk. Weight, hip height, wither height, heart girth, and body length were measured weekly. Weekly plasma samples were analyzed for PUN, NEFA, and glucose. Calves were processed to estimate body composition. Feed efficiency and ADG were greatest for calves fed MILK, least for calves fed 21/21, and intermediate for calves fed 29/16 and 27/33. Calves fed 27/33 or MILK had the greatest gains of fat and percentage fat in the empty body. Body fat percentage of calves fed 29/16 or 21/21 was not changed by diet. Performance of calves fed 27/33 and 29/16 was similar except that calves fed 29/16 were leaner and calves fed 27/33 had a propensity for elevated NEFA. Feeding 180g of CP in the MR was beneficial to calf performance compared with diet 21/21. In experiment two, tissues from a subset of calves [21/21 (n=4), 27/33 (n=5), 29/16 (n=5), MILK (n=3), baseline (n=2)] were scanned using dual energy x-ray absorptiometry to estimate mass, fat, CP, and ash. Liver, organ, and carcass mass by DXA were correlated to scale weights (R²⁺= 0.99, 0.62, and 0.79, respectively). DXA was a poor predictor of percentage fat, CP, and ash (adjusted R²⁺ <0.10). Experiment three determined level of calf mortality in the United States; and identified opportunities to reduce mortality. Herds (n=88) were representative of the US Jersey population. Production averaged 7180 ± 757 kg milk annually. Herds averaged 199 births annually. Mortality was 5.0% from birth to 24 h (M24) of life and 6.7% from 24 h to 3 mo of life (M3). Level of mortality (M24) was highest in herds that calved on pasture. Lower levels of mortality (M3) were associated with use or maternity pens and earlier weaning. / Ph. D.

Two experiments were conducted using newborn colostrum-deprived calves to establish the distribution of uptake of ¹²⁵I- globulin in smail intestine and to investigate effects of added microorganisms on ¹²⁵I-gamma globulin uptake. Ten calves less than 12.5 h of age (X̅ = 7 h) were anesthetized and intestines exteriorized through an abdominal incision. Intestine was ligated into 10 cm segments at 70 cm intervals beginning at the ileocecal junction, injected with ¹²⁵I-gamma globulin in an electrolyte solution and incubated for 1.5 h. One additional segment was formed adjacent to segments 1, 5 and 10 to assess effects of .5 h exposure to ¹²⁵I-gamma globulin on uptake by epithelium. After prescribed gamma globulin exposure, segments were excised, volume of lumen contents, segment weight and tissue activity were determined. Age, birth weight and intestine length were recorded. Location of each segment (PSEG) was expressed as percentage of distance from cecum to abomasum. Uptake was expressed as milligrams gamma globulin internalized per gram of segment tissue. Distribution of gamma globulin uptake after 1.5 h exposure was a cubic function of PSEG. Uptake was greatest In a region 15% of cecumabomasum distance, declining progressively towards the abomasum. After .5 4 exposure, regression of uptake on PSEG was a quadratic function with greatest uptake at 30% of cecum-abomasum distance. Uptake after 1.5 h exposure was greater than after .5 h. In experiment II, 10 calves less than 14 h of age (X̅ = 8.6 h) were anesthetized and intestines surgically exteriorized. Intestine was ligated into segments 10 cm in length at three cm intervals beginning 1.8 m above the ileocecal junction. Seven treatments were assigned in random order to segments in three successive sections of small intestine. Three treatments compared uptake in segments receiving one ml of either live intestine origin bacteria culture, sterile microbiological broth or autoclaved bacteria culture with four h incubation followed by 1.5 h exposure to ¹²⁵I-gamma globulin. Two treatments measured anaerobic microbial growth after four h incubation with one ml of either sterile broth or live bacteria culture. Residual ¹²⁵I-gamma giobulin was measured in segments receiving one ml of sterile broth or live bacteria culture with 5.5 h incubation followed by 15 second exposure to ¹²⁵I-gamma globulin. Measurements were as described for the first 10 calves. Serum corticosteroids, total protein and protein components were measured at O h and 5.5 h later. Uptake was lowest in segments receiving live bacteria as compared to segments receiving sterile inocula. Number of bacteria per gram of segment tissue was negatively correlated with uptake. Low serum corticosteroids were associated with low gamma globulin uptake. Body weight and age were not related to uptake in either experiment.in a decisive manner. / Ph. D.

M. S.

The beef cattle marketing structure imposes stress on calves due to weaning, transport, commingling, and adaptation to new diets, resulting in a weakened immune systems at the height of disease risk, frequently causing bovine respiratory disease. Backgrounding programs facilitate opportunities for calves to overcome stressors by building immunity, and adapting the rumen to high concentrate diets for improved feedlot performance. Four experiments were conducted to compare backgrounding strategies and effects of supplementation frequency performance and the effects of the ruminal environment. In Exp. 1, 48 weaned steers were used to investigate the effects of transportation and supplementation frequency, while in Exp. 2, 36 heifers were used to investigate only supplementation frequency. No differences in gains were observed due to transportation stress or supplementation frequency. Weaning stress resulted in elevated (P < 0.05) creatine kinase and neutrophil:lymphocyte ratios during the first week. In Exp. 3, 48 calves were used to compare the effect of tall fescue type on performance and health. Calves on novel endophyte fescue had higher ADG (P = 0.07) than on endophyte-infected fescue. Experiment 4 investigated the changes in ruminal environment due to supplementation frequency. No differences were observed between supplementation frequencies for ruminal pH, ammonia, or VFA concentration, and DM, or CP digestibility. Therefore, the rumen maintained a hospitable environment to promote bacterial protein synthesis and fiber digestion with every 48 h supplementation. Backgrounding calves with high fiber co-product supplements or on novel endophyte fescue can enhance calf performance. / Master of Science

Two calf feeding trends are emerging in the dairy industry in the United States. Large herds often find it economical to feed pasteurized waste milk; while smaller herds are embracing technological advancements by utilizing automated calf milk feeders. Housing of calves varies depending on feeding mechanism. Calves fed using autofeeders are grouped together but large herds often find it more labor efficient to house calves individually in elevated wooden crates or polyethylene hutches. Two studies were conducted. The objective of the first field study was to evaluate the influence of diet and housing type on growth and morbidity in 84 Holstein heifer calves in a 2 by 2 factorial experimental design. Calves were housed in either polyethylene hutches or elevated wooden crates with slatted floors. Diets consisted of pasteurized waste milk or the same waste milk supplemented to provide approximately 454 g of milk replacer solids containing 25% protein and 10% fat (LOL Balancer). Calves were randomly placed in 1 of 4 treatment groups 48 h after birth and monitored until weaning (~60 d of age). Body weights and hip heights were measured at time of enrollment and weaning. Milk samples of pasteurized waste milk were obtained five times weekly to measure standard bacteriological plate count, fat, protein and total solids content. All calves were fed 3.3 L of liquid diet via bottle at 0730 and 1530 h. Calves were monitored daily for respiratory and digestive illness and treated according to established protocols. Pasteurized waste milk contained 332,171 ° 733,487 cfu/ mL, 3.51 ° 0.59% fat, 3.13 ° 0.30% protein, and 11.64 ° 1.05% total solids. Housing (P = 0.02) and diet (P = 0.01) affected weight gain, but there was no interaction. Least squares average daily gain for crate and hutches were 0.52 ° 0.024 and 0.59 ° 0.024 kg/d. Least squares average daily gain for waste milk and balancer diets were 0.52 ° 0.024 and 0.60 ° 0.024 kg/d, respectively. Housing or diet did not affect hip height growth/d (0.196 ° 0.007 cm). Health of the calves was not affected by diet or housing. Supplementing waste milk with balancer or housing calves in hutches resulted in higher weight gain. The objective of the second study was to evaluate management, and sanitation and consistency of liquid delivered to calves via automated feeders. Ten herds in Virginia and North Carolina with sophisticated (Förster-Technik, Germany) and basic (Biotic Industries Inc., TN, USA) machines completed a 60-question survey concerning calf and autofeeder management. Duplicate milk replacer samples were obtained to measure sanitation, dry matter, and temperature of milk in the autofeeder at the time of the survey. Six dairies from the original 10 were visited monthly for 3 mo for continued evaluation of sanitation, dry matter, and temperature of milk replacer from the autofeeder. Seven herds utilizing basic machines had a mean SPC of 6,925,000 ° 7,371,000 cfu/ml. The mean dry matter and temperature readings were 12.0 ° 2.1 Brix and 38.8 ° 6.7 °C, respectively. Three dairies that used sophisticated autofeeders had a mean SPC of 1,339,000 ° 2,203,000 cfu/ml. Mean dry matter and temperature readings were 10.37 ° 1.68 Brix and 38.6 ° 6.76°C, respectively. Dairies were also categorized based on management strategies. Producers that purchased autofeeders to manipulate feeding rates, refocus labor to sanitation, and care and well-being of calves, or for technological advancements were successful at rearing calves via autofeeders. Dairy producers who purchased an autofeeder to explore feeding options were not as successful because proper time and management was not dedicated to care of calves or to maintenance of the autofeeder. / Master of Science

A five point radial response surface design supplemented with a control group and several single factor points was utilized to investigate the effects of sodium bicarbonate and disodium phosphate on animal performance, ruminal fermentation, ration digestibility and rate of passage of liquid and particulate phases of digesta. Rations contained 0 to 4.5% sodium bicarbonate and 0 to 2.5% disodium phosphate.. Calves were randomly assigned to experimental rations, eight to the control group, six to the centerpoint, two to the radial points and three to the single factor points with restrictions as to time. Animals were on a growth trial from wk 7 to 14. Human samples were taken for determination of pH, buffering capacity, volatile fatty acids anct ruminal ammonia and blood samples for determination of glucose and plasma urea nitrogen at 8, 11 and 14 wk. Rate of passage was determined using ytterbium and cobalt EDTA to follow the particulate and liquid phases of digesta respectively. A digestion trial was carried out concurrently with rate of passage determinations. A two compartment model and a gamma time-dependent model were fitted to marker excretion data. Average daily gains were not significantly affected by buffer additions, but appeared to be maxillial at 1 to 2% sodium bicarbonate and 0% disodium phosphate. Average daily gain and feed efficiency were not changed. Buffers consistently elevated total concentration of volatile fatty acids and osmolality in the rumen. Proportion of volatile fatty acids and acetate to propionate ratio were not changed by buffer addition. Digestibility of dry matter, organic matter, neutral detergent fiber, starch and acid detergent fiber are unchanged. Sodium bicarbonate increased ash digestibility while disodium phosphate tended to decrease ash digestibility. Urine volume and pH were increased by both buffers. Rumen dilution rate for liquid digesta was increased by 50% and particulate digesta increased by 25% with addition of buffers. Flow of liquid digesta was best described by a two compartment model and particulate digesta by a gamma time-dependent model. No benefit of manipulating rate of passage in young dairy calves was observed in this study. Manipulation of ruminal fermentation by rate ot passage has potential to increase; milk production by bypassing starch and protein. Rates of flow of particulate digest may exert important affects on the ruminal acetate to propionate ratio and has potential to explain milk fat depression. / Ph. D.

Mortality and decreased weight gain resulting from infection and disease in dairy calves is a problem within the dairy industry. Colostrum is the sole source of maternal immunity for the calf, having a substantial impact on health and survival. To date, colostrum quality is determined by concentration of antibodies. Colostrum also contains proteins and cells, which may enhance immune development in the neonate. Our goals were to determine the impact of colostral immune cells on (1) immune status during the first month of life and (2) immune development over time. To determine the impact of adoptively transferred colostral immune cells, calves were fed either whole colostrum (WC) or cell-free colostrum (CFC) at birth. During the first month of life, calves fed CFC had decreased numbers of CD4+ T cells when compared to WC-fed calves. However, CFC-fed calves had a greater percentage of monocytes during the first month of life. To determine the influence of colostral immune cells on immune development, cellular blood parameters were measured in response to two series of vaccinations (A and B). After vaccination series A, CFC-fed calves had decreased numbers of B cells when compared to WC-fed calves. After vaccination series B, CFC-fed calves had decreased levels of interleukin-2 gene expression and numbers of CD4+ and gamma delta T cells when compared to WC-fed calves. This study demonstrates that colostral immune cells impact immune status and development in dairy calves. / Master of Science