Biomedical subjects

J S Radcliffe

Publications and source records attributed to J S Radcliffe.

10 recordsLinked to original sources

PubMed2006

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↗

PubMed2006

Estimating equivalency values of microbial phytase for amino acids in growing and finishing pigs fitted with steered ileo-cecal valve cannulas.

Ten crossbred barrows (48.3 +/- 2.3 kg of initial BW) fitted with steered ileo-cecal valve cannulas were used to investigate the effects of supplemental microbial phytase on the apparent ileal digestibilities (AID) of AA, Ca, P, N, and DM, and the apparent total tract digestibilities of Ca, P, N, and DM. All diets were corn-soybean meal-based, and contained 0.44% Ca and 0.40% total P. Diets 1, 2, and 3 contained 12.0, 11.1, and 10.2% CP, respectively. Diets 4 and 5 had the same ingredient composition as diet 3, plus 250 and 500 U/kg phytase (Natuphos), respectively. Pigs were randomly allotted to 1 of 5 dietary treatments in a paired 5 x 5 Latin square with an extra period to test for carryover effects. Each 14-d period consisted of a 7-d adjustment followed by a 3-d total collection, a 12-h ileal digesta collection, a 3-d readjustment, and a second 12-h ileal digesta collection. Pigs were housed individually in metabolism pens (1.2 x 1.2 m). Water was supplied ad libitum, and feed was supplied at a level of 9% of the metabolic BW (BW(0.75)) per day in 2 equal daily feedings. As the dietary CP concentration increased, the AID of CP and all AA measured increased linearly (P < 0.05) with the exception of proline. In addition, the apparent total tract digestibilities (grams per day) and retention of N (grams per day) increased linearly (P < 0.01) with increasing CP levels. Supplementing diets with phytase increased the AID of Ca (P < 0.01), P (P < 0.001), CP (P = 0.07), and the AA (P < 0.10) Gly, Ala, Val, Ile, Thr, TSAA, Asp, Glu, Phe, Lys, and Arg. Protein and phytase response equations were generated for those AA affected (P < 0.10) by both CP level and phytase supplementation. Based on these equations, 500 U/kg of phytase can replace 0.52 percentage units of the dietary CP, which includes a 0.03 percentage unit improvement in Lys AID. The results of this study show that supplementing pig diets with microbial phytase improves CP and AA digestibilities in addition to Ca and P digestibilities.

6-Phytase↗

PubMed2005

Technical Note: Improved technique for fitting pigs with steered ileocecal valve cannulas.

Collection of ileal digesta to evaluate AA digestibilities has become increasingly important in swine nutrition research. Steered ileocecal valve cannulation of pigs permits total collection of ileal digesta, while still allowing normal digesta flow during noncollection periods. This technique was modified and used with 64 crossbred barrows in five trials. Our procedural changes included preoperative i.v. administration of a broad-spectrum antibiotic and nonsteroidal antiinflammatory drug, sharp incision through the muscle layers of the laparotomy wound, use of a heparinized saline lavage solution, replacement of the guide ring with a stylette, and fixing the outer cannula barrel in place with a hose clamp. The current technique involves a right flank laparotomy, parallel and distal to the last rib, with the pig under general anesthesia. A stainless-steel ring (inner ring = 2.0 mm thick, 35.0 mm i.d.) is introduced into the ileal lumen through an enterotomy proximal to the origin of the ileocecal fold. A nylon string attached to this ring is threaded through the ileum and ileocecal valve into the cecum using a silastic stylette, which encases the string. A second stainless-steel ring (outer ring = 2.0 mm thick, 34 mm o.d.) is fixed in place around the ileum, distal to the inner ring and just proximal to the ileocecal valve. A polyurethane cannula barrel (barrel = 100 mm long, 26 mm i.d., 32 mm o.d.; flange = 70 mm o.d.) is introduced into the cecal lumen via an enterotomy through the lateral cecal band and secured in place with two purse-string sutures. The cannula is exteriorized through an incision caudal and proximal to the intial laparotomy site, where it is plugged using a cylindrical stopper (26 mm o.d., 55 mm long) and held in place by a second cannula barrel (barrel = 43 mm length, 33 mm i.d., 41 mm o.d.; flange = 80 mm o.d.). Procedural changes decreased postsurgical complications, as evidenced by decreased seepage around the cannula and fewer and less severe adhesions noted at necropsy. Based on five trials, this technique is a reliable means of collecting ileal digesta for nutrient analyses.

Animals↗

PubMed2004

Animal management to reduce phosphorus losses to the environment.

Water quality in the United States is threatened by contamination with nutrients, primarily nitrogen and phosphorus. Animal manure can be a valuable resource for farmers, providing nutrients, improving soil structure, and increasing vegetative cover to decrease erosion potential. At the same time, application of manure nutrients in excess of crop requirements can result in environmental contamination. Environmental concerns with P are primarily associated with pollution of surface water (streams, lakes, rivers). This pollution may be caused by runoff of P when application to land is in excess of crop requirements. Increased specialization and concentration of livestock and crop production has led to the net export of nutrients from major crop-producing areas of the country to areas with a high concentration of animal agriculture. Concentrated animal agriculture has been identified as a significant source of P contamination of surface water. Areas facing the dilemma of an economically important livestock industry concentrated in an environmentally sensitive area have few options. If agricultural practices continue as they have in the past, continued damage to water resources and a loss of fishing and recreational activity are inevitable. If agricultural productivity is decreased, however, the maintenance of a stable farm economy, a viable rural economy, and a reliable domestic food supply are seriously threatened. Decreasing the P content of manure through nutrition is a powerful, cost-effective approach to reducing P losses from livestock farms and will help farmers meet increasingly stringent environmental regulations. This paper reviews opportunities available to reduce the P content of livestock manure, including more accurate interpretation of the published P requirements of animals, improved diet formulation and group-feeding strategies to more precisely meet requirements, and approaches to improve availability of feed P for monogastric and ruminant species.

6-Phytase↗

PubMed2004

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↗

PubMed2000

Comparison of genetically engineered microbial and plant phytase for young broilers.

This study was conducted to compare the efficacy of genetically engineered microbial (Natuphos) and plant (Phytaseed) phytase for enhancing the utilization of phytate P in corn-soybean meal-based diets fed to young broilers and to evaluate the safety of Phytaseed phytase. Three levels of each of the two sources of phytase (250, 500, and 2,500 U/kg of diet) were added to a corn-soybean meal basal diet containing 0.46% total P, 0.21% nonphytate P, and 0.92% Ca. There were eight cages per treatment (eight birds per cage for Weeks 2 to 3 and seven birds for Weeks 4 to 5), except for the basal diet without added phytase that had 16 cages. Cage BW and feed consumption were recorded weekly. During Week 5, cage excreta samples were collected for determination of apparent retention coefficients of DM, Ca, and P. At the end of Week 5, all birds were killed, and the left and right toes were removed for determination of toe ash weight and percentage. Forty birds (one per cage from the diet without added phytase and diets with 500 or 2,500 U phytase/kg from both sources) were randomly selected for gross necropsy and histologic evaluation of liver, kidney, and bone tissues. Addition of both sources of phytase resulted in similar increases (P < 0.05) of BW gain; feed intake; gain:feed; apparent retention of DM, P and Ca; and toe measurements. Phosphorus excretion decreased as phytase addition increased. No significant abnormalities were seen in any of the 40 broilers necropsied. Further, the fit of a nonlinear function revealed that most measurements reached a plateau at 2,500 U/kg. Based on performance, bone characteristics, and retention of P, Ca, and DM of young broilers, the efficacy of Phytaseed phytase was similar to that of Natuphos phytase for enhancing the utilization of phytate P in corn-soybean meal-based diets. General necropsy and histologic examination of liver, kidney, and tibial tissues revealed no adverse effects of phytase source or level.

6-Phytase↗

PubMed2000

Comparison of phytase from genetically engineered Aspergillus and canola in weanling pig diets.

Ninety-six crossbred pigs with an average weight of 9.0 kg were used in a 5-wk trial to compare the efficacy of genetically engineered Aspergillus ficuum phytase, expressed in Aspergillus niger (Natuphos) or in canola seed (Phytaseed), for enhancing the utilization of phytate P in corn-soybean meal-based diets fed to young pigs and to evaluate the safety of Phytaseed phytase. Three levels of the two sources of phytase (250, 500, or 2,500 U/kg of diet) were added to a corn-soybean meal basal diet containing .35% total P, .09% available P, and .50% Ca. There were six pens per treatment (one barrow and one gilt/pen), except that the diet without added phytase was fed to 12 pens of pigs. Pen feed consumption and BW were recorded weekly. During wk 5, pen fecal samples were collected for determination of apparent digestibilities of DM, Ca, and P. At the end of wk 5, all barrows were killed, and the 10th rib on both sides was removed for determination of shear force and energy. Thirty pigs (six from the diet without added phytase and the diets with 500 and 2,500 U/kg phytase from both sources) were randomly selected for gross necropsy and histologic evaluation of liver, kidney, and bone tissues. Both sources of phytase were equally effective in increasing (P < .05) daily gain, gain:feed, apparent digestibilities of DM, P, and Ca, and 10th rib measurements. Fecal P excretion was reduced with phytase addition. Feed intake was increased by phytase levels during wk 4 to 5. No significant abnormalities were seen in any of the 30 pigs necropsied. The fit of a nonlinear function revealed that most measurements were reaching a plateau at 2,500 U/kg phytase. In summary, based on performance, bone measurements, and digestibilities of P, Ca, and DM of young pigs, the efficiency of Phytaseed was similar to that of Natuphos for enhancing the utilization of phytate P in corn-soybean meal-based diets. General necropsy and histologic examination of tissues indicated no toxic effect of phytase.

6-Phytase↗

PubMed1998

The effects of microbial phytase, citric acid, and their interaction in a corn-soybean meal-based diet for weanling pigs.

Crossbred weanling pigs (an equal number of barrows and gilts) with an average initial weight of 7.4 (Exp. 1) or 9.6 kg (Exp. 2) were used in two 4-wk experiments (Exp. 1, n = 96; Exp. 2, n = 96) to investigate the effects of added phytase or citric acid on performance, rib mineralization, gastric pH, and digestibility measurements. A corn-soybean meal-based diet low in Ca and P was used in both experiments. In Exp. 1, three citric acid levels (0, 1.5, or 3.0%) and four phytase levels (0, 250, 500, or 750 U/kg) were used in a 3 x 4 factorial arrangement of treatments. In Exp. 2, two citric acid levels (0 or 2.0%) and three phytase levels (0, 250, or 500 U/kg) were used in a 2 x 3 factorial arrangement of treatments. Phosphorus was maintained at .33 and .34% in Exp. 1 and 2, respectively. Calcium was maintained at a 2.5:1 ratio with total available P (available P plus the estimated released phytate P by phytase) in Exp. 1 and at a level of .44% in Exp. 2. In both experiments, BW and feed consumption were measured weekly, and pen fecal samples were collected twice daily for 5 d during wk 4. At the end of wk 4, the barrow in each pen was killed following a fast-refeed-fast (22-1-2 h) regimen for collection of 10th ribs and stomach digesta. In Exp. 1 and 2, phytase addition did not affect (P > .05) performance but linearly increased (P < .05) rib shear force, shear energy, dry bone weight, ash weight, ash percentage, and Ca and P digestibilities. Addition of citric acid in both experiments reduced dietary pH and stomach digesta pH (P < .05). The addition of citric acid improved (P < .05) ADG, feed efficiency, and Ca digestibility in Exp. 1, but it had no effect on performance and Ca digestibility in Exp. 2. In summary, the additions of citric acid and phytase to weanling pig diets were each beneficial, but no synergistic effects were observed.

6-Phytase↗

PubMed1997

Factors affecting phosphorus and calcium digestibility in diets for growing-finishing pigs.

In two experiments, we investigated various factors that affect the estimation of the apparent total tract digestibility (ATTD) of DM, Ca, and total P in diets for growing-finishing pigs. In Exp. 1, the effects of age, housing, and calculation method (indicator [Cr2O3] vs 10-d total collection) were determined. Eighteen barrows and gilts (40 to 95 kg BW) were housed in six pens, and ATTD was estimated using the indicator method. Twelve barrows were housed in metabolic crates, and ATTD was estimated using both calculation methods. Dietary treatments were 1) a tapioca-soybean-barley-based diet, 2) Diet 1 supplemented with 400 FTU microbial phytase/kg of diet, and 3) a corn-soybean meal-based diet: In Exp. 2, six barrows (95 to 120 kg BW) were fed a phytase-deficient diet to investigate the effects of coprophagy (40 g fresh feces/kg of diet) and movement. Pigs were fed at 2.8 times maintenance requirement (418 kJ ME/BW.75); water supply was 2.5 L/kg of feed. The ATTD increased as BW increased. Phytase enhanced total P ATTD by an average of 18.1 percentage units. The ATTD of DM was higher and the ATTD of Ca and total P (P < .001) were lower in pigs housed in pens than in pigs housed in metabolic crates. Fecal consumption and movement led to numerical increases in Ca (P = .217) and total P (P = .103) ATTD. Estimates of Ca and total P ATTD using pigs in metabolic crates are lower than estimates in practice.

6-Phytase↗

PubMed1997

The effects of sow parity on digestibility of proximate components and minerals during lactation as influenced by diet and microbial phytase supplementation.

Ninety-six (Finnish Landrace x Dutch Landrace) reproductive sows were used at parities 1, 3, 5, and 7 + 8 from d 107 of gestation to d 21 of lactation to investigate the effects of diet and parity on apparent total tract digestibility (ATTD). Animals were randomly assigned to one of four dietary treatments. Dietary treatments were 1) a P-deficient (1.1 g digestible P [dP]/kg) Dutch semipractical diet; 2) Diet 1 supplemented with 400 FTU Aspergillus niger phytase per kilogram of diet (1.7 g dP/kg); 3) a corn-soybean meal-based diet (1.3 g dP/kg); and 4) Diet 3 supplemented with extra monocalcium phosphate (MCP; 2.4 g dP/kg of diet). Animals were fed twice daily at 2.8 times maintenance (418 kJ ME/ BW75) from d 8 to the end of lactation. Feces and urine were collected during d 11 to 13 and d 18 to 20 of lactation. The ATTD of DM, OM, ash, CP, Ca, Mg, and total P (P < .01) were higher for the corn-soybean meal-based diets than for the Dutch semipractical diet not supplemented with phytase. Addition of MCP enhanced total P ATTD by an average of 6.7 percentage units. Addition of microbial phytase improved Ca, Mg, and total P ATTD, but the effects were dependent on the stage of lactation. Lower ATTD of OM and CP were seen for first parity animals compared with higher parity sows. The ATTD of Mg increased with increasing parity. Parity had little effect on the ATTD of minerals during lactation, and dietary effects were prominent and followed a similar trend to those seen in growing-finishing pigs.

6-Phytase↗