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Biomedical subjects

K E Griswold

Publications and source records attributed to K E Griswold.

15 recordsLinked to original sources

Effects of replacing sulfate with hydroxychloride sources of trace minerals on mineral status and production performance in dairy cows.

The objectives were to determine the effects of replacing sulfate with hydroxychloride sources of Cu, Mn, and Zn on mineral status and production performance in dairy cows. One hundred forty-one Holstein cows were stratified by parity group prepartum as nulliparous (lactation 0) or parous cows (lactation >0) and, within parity, cows were blocked by genomic breeding value for ECM yield (nulliparous cows) or recently completed lactation 305-d ECM (parous cows) and then randomly assigned to 1 of 2 treatments. Treatments were supplemental sources of Cu, Mn, and Zn as sulfate trace minerals (STM) or hydroxychloride trace minerals (HTM). Diets were formulated to contain approximately 16, 60, and 60 mg/kg of Cu, Mn, and Zn, respectively, and treatments were fed from 246 d of gestation to 105 d of lactation. Cows were weighed twice weekly prepartum and intake of DM, milk yield, and postpartum BW were measured daily, and composition of milk was analyzed twice weekly. Blood was sampled pre- and postpartum and hepatic tissue collected at 10 (50 STM and 52 HTM cows) and at 50 d postpartum (17 STM and 18 HTM cows) and analyzed for concentrations of minerals. Treatment did not affect intake or measures of energy balance prepartum. Numerical results between parentheses are presented following the sequence of STM and HTM. Concentrations of trace minerals in serum differed between treatments only prepartum and those of Cu (1.291 vs. 1.183 ± 0.031 mg/L) and Zn (1.307 vs. 1.211 ± 0.031 mg/L) were greater for cows fed STM compared with cows fed HTM; however, the opposite response was observed for serum Mn (1.628 vs. 1.754 ± 0.038 µg/L). Treatment did not affect liver Mn or Zn concentration, but for Cu, cows fed STM had greater concentration in the liver on d 10 postpartum compared with cows fed HTM (310 vs. 296 ± 7 mg/kg DM); however, the opposite response was observed on d 50 and cows fed STM had smaller concentration of Cu in liver than those fed HTM (287 vs. 318 ± 9 mg/kg DM). Cows fed HTM produced an additional 1.0 kg colostrum than STM cows (5.21 vs. 6.22 ± 0.63 kg), thus resulting in increased yield of colostrum solids. Treatment did not affect the content or yield of IgG in colostrum. Cows fed HTM produced 1.3 kg/d more milk (41.7 vs. 43.0 ± 0.5 kg/d) and 1.5 kg/d more energy-corrected milk (42.5 vs. 44.0 ± 0.6 kg/d) in the first 15 wk of lactation compared with cows fed STM. The estimated NEL content of the postpartum diet consumed by cows, after accounting for the different energy sinks and DMI, was 3.4% greater for HTM than STM (1.68 vs. 1.74 ± 0.02 Mcal/kg). Replacing sulfate with hydroxychloride sources of Cu, Mn, and Zn had small effects on the concentrations of those minerals in tissues and improved production performance in the first 15 wk of lactation.

Animals↗

The digestive fate of Escherichia coli glutamate dehydrogenase deoxyribonucleic acid from transgenic corn in diets fed to weanling pigs.

Corn containing genetically engineered plasmid DNA encoding an Escherichia coli glutamate dehydrogenase (gdhA) was fed to 19-d-old weanling swine to trace the digestive fate of the transgenic DNA. Eight pens of 8 pigs were fed a commercial (nongdhA) starter for 2 wk. One pig was randomly selected from each pen for 0-h control samples. The remaining 56 pigs were transitioned onto a corn-soybean meal diet and fed a diet containing 58% gdhA corn for approximately 1 wk; immediately thereafter, liver, 10th rib muscle, white blood cells, and plasma from the hepatic portal vein and ingesta from the stomach, distal ileum, and large intestine were collected. The DNA was extracted and the concentration determined via spectrophotometry. Polymerase chain reaction and gel electrophoresis were performed with primers designed to amplify 490 bp that included the plasmid's ligation site between the maize ubiquitin and the gdhA genes. The gdhA corn-derived DNA and diet served as positive assay controls, and conventional corn DNA and distilled water acted as negative assay controls. Detection limits were 0.99 fg of target DNA confounded with 500 ng of conventional corn DNA per each 20 &L reaction. Transgenic DNA was detected in 71.43% of the stomach and 1.79% of the ileal ingesta samples from treatment animals but was not detected in the large intestine, white blood cells, plasma, liver, or muscle samples. Transgenic DNA was not detected in any sample from 0-h control animals. Stomach and ileal ingesta samples were further analyzed using real-time PCR. With an estimated limit of detection of 1.049 ag/microL, 89.29% of the stomach ingesta samples were positive (average 1.56 fg target DNA). The proportion of transgenic DNA to total DNA differed between diet and stomach ingesta samples (P < 0.001). Despite the greater sensitivity of real-time PCR, target DNA was detected in only 1.79% of ileal ingesta. These data suggest that the gdhA transgene began degradation in the stomach and was nondetectable in the large intestine.

Animal Feed↗

Fecal prevalence and diversity of Salmonella species in lactating dairy cattle in four states.

Salmonella is one of the most serious foodborne pathogenic bacteria in the United States, causing an estimated 1.3 million human illnesses each year. Dairy cows can be reservoirs of foodborne pathogenic bacteria, including Salmonella spp.; it is estimated that from 27 to 31% of dairy herds across the United States are colonized by Salmonella. The present study was designed to examine the occurrence of Salmonella spp. on dairies and to examine the serotypic diversity of Salmonella isolates on sampled dairies from across the United States. Fecal samples (n = 60 per dairy) were collected from 4 dairies in each of 4 states for a total of 960 fecal samples representing a total population of 13,200 dairy cattle. In the present study, 93 of 960 samples (9.96%) collected were culture-positive for Salmonella enterica. At least one Salmonella fecal-shedding cow was found in 9 of the 16 herds (56%) and the within-herd prevalence varied in our study from 0% in 7 herds to a maximum of 37% in 2 herds, with a mean prevalence among Salmonella-positive herds of 17%. Seventeen different serotypes were isolated, representing 7 different Salmonella serogroups. There were 2 or more different serogroups and serotypes present on 7 of the 9 Salmonella-positive farms. Serotypes Montevideo and Muenster were the most frequent and widespread. From our data, it appears that subclinical colonization with Salmonella enterica is relatively common on dairy farms and is represented by diverse serotypes on US dairy farms.

Animals↗

Nutritional value of a corn containing a glutamate dehydrogenase gene for growing pigs.

Eight female PIC Line 42 pigs (initial BW = 47.5 +/- 1.8 kg) were used in a two-period switchback design (n = 4 per treatment per period) to evaluate the nutritional difference between a genetically modified corn and a similar nontransgenic corn. The genetically altered corn (gdhA+) contained a glutamate dehydrogenase gene isolated from Escherichia coli. The non-transgenic corn was the same variety lacking the transgenic cassette, grown at the same two locations. Pigs were surgically fitted with steered ileocecal valve cannulas for collection of ileal digesta. Diets were made up of primarily one of the two corn sources. Dietary AA profiles were adjusted using crystalline AA to match Illinois Ideal Protein Ratios. Pigs were limit-fed at 8% of metabolic body weight (BW0.75) in two equal feedings at 0600 and 1800 daily throughout the experiment. The study consisted of two 15-d periods. Each period consisted of a 7-d acclimation period, a 3-d total collection of feces and urine, two 12-h ileal collections, and a 3-d adjustment period between ileal collections to ensure adequate hydration. Crude protein, leucine, methionine, alanine, aspartic acid, glutamic acid, and tyrosine concentrations were greater (P < 0.05) in the gdhA+ corn than in the nontransgenic variety. The presence of the gene did not alter (P > 0.17) BW gain. Similarly, DM digestibility, fecal N excretion (grams per day), apparent total-tract N digestibility, N balance, net protein utilization, and N retained as percentages of absorbed were not affected (P > or = 0.32) by the gene modification. Apparent ileal AA digestibility values did not differ (P > 0.31) between the two dietary treatments. Results of this study suggest corn that contains the E coli. gene for glutamate dehydrogenase was nutritionally equivalent to the unaltered variety.

Animal Feed↗

Effects of substrate, passage rate, and pH in continuous culture on flows of conjugated linoleic acid and trans C18:1.

A dual-flow continuous culture system consisting of 4 fermenters was used in a 4 x4 Latin square design. The objective of the research was to evaluate the effects of solid dilution rate (SDR), pH, and concentration of linoleic acid (LA) in the feed mixture on the production of conjugated linoleic acid (CLA) and trans-C18:1. The 4 treatments were 1) control = pH 6.5, 1% LA, 4%/h SDR; 2) high solid dilution rate (HSDR) = pH 6.5, 1% LA, 8%/h SDR; 3) high linoleic acid (HLA) = pH 6.5, 3% LA, 4%/h SDR; and 4) low pH (LPH) = pH 5.8, 1% LA, 4%/h SDR. Inoculum was collected 6 h after feeding from a cow fed 40% alfalfa hay and 60% grain. Liquid dilution rate was held at 0.12/h. All treatments except HLA contained 2% tallow. The LA was dissolved in buffer and continuously infused into the fermenters. The CLA flows were 16.5, 20.4, 23.2, and 25.2 mg/d for control, HSDR, HLA, and LPH, respectively. Compared with control, LPH increased flows of CLA, cis-C18:1, and C18:2, and decreased flow of C18:0. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibilities were not affected by pH. The HSDR tended to increase CLA flow compared to control, possibly because a shorter solid retention time led to incomplete biohydrogenation (BH). The NDF and ADF digestibilities and bacterial numbers were reduced by HSDR. With more LA available as a substrate for CLA, HLA resulted in a higher flow of CLA than control. The HLA resulted in the highest acid detergent fiber and fatty acid digestibilities, bacterial numbers, and BH. Increasing solids passage rate, reducing pH, and increasing dietary LA appears to increase in vitro CLA production.

Acetates↗

Domperidone can ameliorate deleterious reproductive effects and reduced weight gain associated with fescue toxicosis in heifers.

The aim of this study was to evaluate the effect of a dopamine antagonist, domperidone, in nonpregnant, reproductively cycling heifers consuming endophyte-infected (EI) fescue diets. Thirty crossbred heifers (Angus x Holstein or Hereford x Holstein) were assigned to one of three treatment groups (n = 10); endophyte-free (EF) fescue diet, EI fescue diet, or endophyte-infected diet and treated with domperidone (EID). Heifers fed EI diets had decreased weight gains compared with heifers fed EF or EID (P < 0.05) during a 21-d treatment period. Ovarian structures were monitored via transrectal ultrasound to determine follicle size and day of ovulation. Blood plasma samples were collected daily and analyzed for progesterone concentration to determine luteal function. Heifers ingesting EI diets had estrous cycles of shorter duration and lower mid-cycle progesterone concentrations than heifers in the EF or EID treatments (P < 0.05). Ovaries from a subset of heifers in each group (n = 3 per group) were harvested and in vitro secretion of progesterone from luteal tissue extracts was determined. No differences in progesterone concentrations were detected among luteal tissue incubates (P > 0.05). These results suggest that domperidone supplementation of heifers consuming EI fescue may ameliorate certain symptoms of fescue toxicosis.

Acremonium↗

Effects of urea infusion and ruminal degradable protein concentration on microbial growth, digestibility, and fermentation in continuous culture.

The effects of urea and rumen-degradable protein (RDP) on microbial growth, digestibility, and fermentation were examined using dual-flow continuous culture. The experimental design was a 4 x 4 Latin square with a 2 x 2 factorial arrangement of treatments. Factors were urea infusion (0.4 g/L of artificial saliva) and RDP concentration, and the treatments were as follows: 1) low RDP (8% of dietary dry matter) without urea (LDNU), 2) high RDP (11% of dietary dry matter) without urea (HDNU), 3) low RDP (8% of dietary dry matter) with urea (LDU), and 4) high RDP (11% of dietary dry matter) with urea (HDU). The LDNU (i.e., negative control) and HDNU treatments were formulated to be nitrogen limiting. Results indicated that infusion of urea increased all digestibility measurements (P < 0.05), which in turn increased (P < 0.05) volatile fatty acid, NH3 nitrogen, trichloroacetic acid-soluble nitrogen, and soluble protein concentrations. Increasing dietary RDP improved dry matter and organic matter digestibility (P < 0.05) but did not alter acid detergent fiber or nonfiber carbohydrate digestibilities (P > 0.05). Isobutyrate concentration decreased (P = 0.05) with increased RDP. Increased dietary RDP increased crude protein degradation and soluble protein concentration (P < 0.05), but NH3 nitrogen, trichloroacetic acid-soluble nitrogen, and peptide nitrogen were unaffected by changing RDP levels. Microbial growth efficiency was 19.9, 24.9, 28.0, and 32.2 g N/g organic matter truly digested for LDNU, HDNU, LDU, and HDU, respectively, and was significantly improved both by urea infusion (P = 0.002) and increased RDP concentration (P = 0.021). The interactions of urea and RDP (P < 0.05) were explained by the high digestibility of neutral detergent fiber, nonstructural carbohydrate, and especially hemicellulose, with the HDNU treatment. The results of this study indicated that hemicellulose-degrading bacteria were able to effectively compete with nonstructural carbohydrate-degrading bacteria for available peptide and amino acid nitrogen. Further, the extent of protein degradation was dependent on the availability of NH3 nitrogen in the system.

Animals↗

Effectiveness of short-term feeding strategies for altering conjugated linoleic acid content of beef.

A steer finishing trial was performed to determine the effect of short-term dietary regimens on conjugated linoleic acid (CLA) content of muscle tissues. The experimental design was an incomplete 3 x 2 factorial, with three levels of soybean oil (SBO; 0, 4, and 8% of diet DM) and two levels of forage (20 vs. 40% of diet DM). Forty Angus x Hereford steers averaging 504 +/- 29.0 kg were allotted randomly to one of four treatments for the last 6 wk of the finishing period. Treatments were: 80:20 concentrate:forage control diet (C); 80:20 concentrate:forage + 4% SBO (C4); 60:40 concentrate:forage + 4% SBO (F4); and 60:40 concentrate:forage + 8% SBO (F8). After 42 d on the experimental diets, steers were sacrificed and samples were collected from the chuck, loin, and round muscle groups. Fatty acid (FA; mg/100 mg of FA) composition was determined by gas-liquid chromatography. Data were statistically analyzed with mixed models procedures. The performance and carcass quality model included the effects of SBO and forage. The model for FA composition included the effects of SBO, forage, muscle group, and interactions. Orthogonal contrasts were used to determine linear effects of SBO. There were no differences in growth performance among treatments (P > 0.05). Increasing dietary SBO linearly decreased dressing percent (P = 0.04), and tended to linearly decrease marbling score (P = 0.12) and quality grade (P = 0.08). The only CLA isomer detected in tissue samples was cis-9,trans-11. Addition of SBO to diets linearly increased linoleic acid (18:2n-6; P = 0.04) and tended to linearly increase linolenic acid (18:3n-3; P = 0.10) in muscle tissues. The CLA in lean tissues was decreased (P = 0.005) with SBO-containing diets. These findings suggest that increased PUFA may limit ruminal production of CLA and trans-vaccenic acid (VA) and/or may depress stearoyl-CoA desaturase expression or activity in lean tissues, which in turn limits CLA formation and accretion in tissues. Increasing dietary forage tended to increase 18:0, 18:2n-6, CLA, and 18:3n-3 (P < 0.15), suggesting that increased forage may mitigate toxic effects of PUFA on ruminal biohydrogenation, thereby increasing the pool of CLA and VA available for CLA formation and accretion in tissues. Short-term feeding of elevated SBO and forage levels can alter FA profiles in muscle tissues.

Animal Feed↗

Proteolytic activities of the starch-fermenting ruminal bacterium, Streptococcus bovis.

The objective of this study was to characterize the extracellular proteolytic activity of Streptococcus bovis. Strains KEG, JB1, NCFB 2476, and K11.21.09.6C produced very similar large molecular weight (160-200 kDa) extracellular proteases that were specifically inhibited by PMSF, a serine protease inhibitor. Further experiments with S. bovis KEG indicated that cultures grown with casein as the sole added N source produced the greatest level of proteolytic activity, and the level of proteolytic activity was independent of growth rate. Clarified ruminal fluid (CRF) decreased proteolytic activity by 54% compared with cultures grown with casein alone, and addition of exogenous peptides and carbohydrates (CHO) to the CRF further reduced the level of proteolytic activity by 44% and 52%, respectively. These results suggested that the proteolytic activity of S. bovis KEG was modulated by available N source and that the proteolytic activity was present for reasons other than providing N for growth. The role of S. bovis in ruminal proteolysis requires further definition, but phenotypic similarity among some ruminal strains would suggest a common niche in ruminal proteolysis. The uniformity of proteolytic activities could make S. bovis a prime candidate for manipulation in ruminal proteolysis control strategies.

Animals↗

Diversity of extracellular proteolytic activities among Prevotella species from the rumen.

The current research was aimed at comparing proteolytic activities among ruminal Prevotella spp. Growth rates of Prevotella sp. 2202, Prevotella ruminicola D31d, P. brevis GA33, P. albensis M384, and P. bryantii B(1)4 varied with N source, and no one N source produced the fastest growth in all species. Proteolytic activity was greatest with casein compared with peptides, AA, and NH(4)Cl in all species. Proteolytic activity of Prevotella sp. 2202, P. brevis GA33, and P. bryantii B(1)4 was modulated by N source. With gelatin co-polymerized SDS-PAGE, the extracellular activities of the Prevotella spp. showed wide variation in number, size, and type of proteases. Prevotella sp. 2202 and P. albensis M384 produced metalloproteases of low molecular weight (40 kDa). P. ruminicola D31d produced one cysteine protease (100-200 kDa) and two metalloproteases (90-100 kDa). P. brevis GA33 generated a diffuse clearing zone (95-160 kDa) containing serine, cysteine, and metalloproteases. P. bryantii B(1)4 produced a metalloprotease greater than 200 kDa in size. The molecular sizes provided are estimations and served only to differentiate among the bacterial species in this study. Large variations in proteolytic activities among species and the known genetic diversity of the Prevotella taxon suggested that targeting this bacterial assemblage for genetic manipulation in order to alter the bacterial impact on ruminal protein degradation would be difficult.

Animals↗

Degradation of protein and utilization of the hydrolytic products by a predominant ruminal bacterium, Prevotella ruminicola B1(4).

Degradation and utilization of protein by Prevotella ruminicola B1(4), a proteolytic bacterium that is prominent in the rumen, was examined. In preliminary experiments, proteinaceous N sources produced faster growth rates than did NH4Cl, based on changes in optical density over time. However, ammonium chloride produced a greater maximum cell density than did proteinaceous N sources. Of the proteinaceous N sources, an enzymatic hydrolysate of soybean protein with a relative peptide size of 3 AA residues produced a greater growth rate and maximum cell density compared with the other proteinaceous N sources. Further experiments revealed that P. ruminicola B1(4) grew faster and to a greater final dry weight with soybean protein than with casein. Degradation of both proteins was low as was indicated by the slow disappearance of soluble protein, low concentrations of free AA and peptides, and the decrease in ammonia concentrations over time. Patterns of degradation did differ between the two proteins, however. Accumulation of peptides and free AA from soybean protein peaked 2 h earlier than those from casein, and concentrations of free AA and peptides from soybean protein were lower on average than those from casein. Prevotella ruminicola B1(4) preferentially utilized Asp, Ile, Leu, Lys, and Arg from soybean protein compared with casein. The relative size of peptides that accumulated from both proteins, as determined by the ratio of ninhydrin reaction after HCl hydrolysis to ninhydrin reaction before HCl hydrolysis, suggested that part of the proteolytic activity of P. ruminicola B1(4) is a dipeptidase. Our findings suggest that P. ruminicola may have a greater impact on peptide degradation than on protein degradation in the rumen.

Amino Acids↗

Effect of form of nitrogen on growth of ruminal microbes in continuous culture.

A study was conducted to determine the effect of various forms of N on the growth of ruminal microbes in a continuous culture system with solids and liquid dilution rates comparable to those of a high-producing dairy cow. Nitrogen forms were isolated soy protein, soy peptides, individual amino acids (AA) blended to profile soy protein, and urea, which were fed alone and in combinations so that the total N provided was 1.6% of the diet DM. The 100% soy protein treatment resulted in reduced digestion of N and nonstructural carbohydrate compared with other N forms, and outflow of bacterial N/24 h was less than when peptides were fed. This suggested that proteolysis rather than peptide uptake was the rate-limiting step in N utilization in this study. Non-urea N forms increased ADF digestion, total VFA production and the molar percentages of isobutyrate, isovalerate, and valerate compared to urea, which reflected the contribution of carbon skeletons of AA. When combinations of N forms were used, each form contributed an equal quantity of N, 50% of the total treatment, which was .8% of the diet DM. Combinations of N forms did not enhance, and in most cases reduced, ADF and NDF digestion when compared with individual N forms, and no combinations increased microbial growth over that of the individual forms. These results confirm that N forms other than ammonia are needed not only for maximum microbial growth, and they further demonstrate a need for non-protein N for the fiber digestion. In addition, results of this study suggest a requirement for a minimum level of peptide or AA N, which was met only when individual N forms were fed.

Amino Acids↗

Amelioration of cardiotoxic effects of alcohol by vitamin E.

Injection of mice with a single ip dose of 2g/kg of ethanol leads to time dependent increases of lactic dehydrogenase plasma isoenzymes indicative of myocardial damage. Electron microscopic analysis of the myocardium shows changes in mitochondrial structure, endoplasmic reticulum and myofibrils. Pretreatment of the animals with 86 units of alpha tocopherol partially prevented the changes in isoenzyme patterns and reduced the electron microscopic evidence of myocardial damage. The study supports previous findings that some of the toxic effects of alcohol might be mediated through free radical mechanisms leading to lipid peroxidation and that the ameliorating effect of alpha tocopherol could relate to its function as antioxidant and free radical scavenger.

Animals↗