Carbapenemase-producing Enterobacterales in hospital drains in Southern Ontario, Canada.

BACKGROUND: Hospital drains may be an important reservoir for carbapenemase-producing Enterobacterales (CPE). AIM: To determine prevalence of CPE in hospital drains exposed to inpatients with CPE, relatedness of drain and patient CPE, and risk factors for drain contamination. METHODS: Sink and shower drains in patient rooms and communal shower rooms exposed to 310 inpatients with CPE colonization/infection were cultured at 10 hospitals. Using short- and long-read whole-genome sequencing, inpatient and corresponding drain CPE were compared. Risk factors for drain contamination were assessed using multi-level modelling. FINDINGS: Of 1209 exposed patient room and communal shower room drains, 53 (4%) yielded 62 CPE isolates in seven (70%) hospitals. Of 49 CPE isolates in patient room drains, four (8%) were linked to prior room occupants. Linked drain/room occupant pairs included Citrobacter freundii ST18 isolates separated by eight single nucleotide variants (SNVs), related blaKPC-containing IncN3-type plasmids (different species), related blaKPC-3-containing IncN-type plasmids (different species), and related blaOXA-48-containing IncL/M-type plasmids (different species). In one hospital, drain isolates from eight rooms on two units were Enterobacter hormaechei separated by 0-6 SNVs. Shower drains were more likely to be CPE-contaminated than hand hygiene (odds ratio: 3.45; 95% confidence interval: 1.66-7.16) or patient-use (13.0; 4.29-39.1) sink drains. Hand hygiene sink drains were more likely to be CPE-contaminated than patient-use sink drains (3.75; 1.17-12.0). CONCLUSION: Drain contamination was uncommon but widely dispersed. Drain CPE unrelated to patient exposure suggests contamination by undetected colonized patients or retrograde (drain-to-drain) contamination. Drain types had different contamination risks.

Neisseria gonorrhoeae Sequence Typing for Antimicrobial Resistance, a Novel Antimicrobial Resistance Multilocus Typing Scheme for Tracking Global Dissemination of N. gonorrhoeae Strains.

A curated Web-based user-friendly sequence typing tool based on antimicrobial resistance determinants in Neisseria gonorrhoeae was developed and is publicly accessible (https://ngstar.canada.ca). The N. gonorrhoeae Sequence Typing for Antimicrobial Resistance (NG-STAR) molecular typing scheme uses the DNA sequences of 7 genes (penA, mtrR, porB, ponA, gyrA, parC, and 23S rRNA) associated with resistance to ß-lactam antimicrobials, macrolides, or fluoroquinolones. NG-STAR uses the entire penA sequence, combining the historical nomenclature for penA types I to XXXVIII with novel nucleotide sequence designations; the full mtrR sequence and a portion of its promoter region; portions of ponA, porB, gyrA, and parC; and 23S rRNA sequences. NG-STAR grouped 768 isolates into 139 sequence types (STs) (n = 660) consisting of 29 clonal complexes (CCs) having a maximum of a single-locus variation, and 76 NG-STAR STs (n = 109) were identified as unrelated singletons. NG-STAR had a high Simpson's diversity index value of 96.5% (95% confidence interval [CI] = 0.959 to 0.969). The most common STs were NG-STAR ST-90 (n = 100; 13.0%), ST-42 and ST-91 (n = 45; 5.9%), ST-64 (n = 44; 5.72%), and ST-139 (n = 42; 5.5%). Decreased susceptibility to azithromycin was associated with NG-STAR ST-58, ST-61, ST-64, ST-79, ST-91, and ST-139 (n = 156; 92.3%); decreased susceptibility to cephalosporins was associated with NG-STAR ST-90, ST-91, and ST-97 (n = 162; 94.2%); and ciprofloxacin resistance was associated with NG-STAR ST-26, ST-90, ST-91, ST-97, ST-150, and ST-158 (n = 196; 98.0%). All isolates of NG-STAR ST-42, ST-43, ST-63, ST-81, and ST-160 (n = 106) were susceptible to all four antimicrobials. The standardization of nomenclature associated with antimicrobial resistance determinants through an internationally available database will facilitate the monitoring of the global dissemination of antimicrobial-resistant N. gonorrhoeae strains.

Risk factors for sporadic domestically acquired Salmonella serovar Enteritidis infections: a case-control study in Ontario, Canada, 2011.

In Ontario, Canada, the number of Salmonella Enteritidis (SE) cases increased over the years 2005-2010. A population-based case-control study was undertaken from January to August 2011 for the purpose of identifying risk factors for acquiring illness due to SE within Ontario. A total of 199 cases and 241 controls were enrolled. After adjustment for confounders, consuming any poultry meat [adjusted odds ratio (aOR) 2·24, 95% confidence interval (CI) 1·31-3·83], processed chicken (aOR 3·32, 95% CI 1·26-8·76) and not washing hands following handling of raw eggs (OR 2·82, 95% CI 1·48-5·37) were significantly associated with SE infection. The population attributable fraction was 46% for any poultry meat consumption and 10% for processed chicken. Poultry meat continues to be identified as a risk factor for SE illness. Control of SE at source, as well as proper food handling practices, are required to reduce the number of SE cases.

Subtyping of a large collection of historical Listeria monocytogenes strains from Ontario, Canada, by an improved multilocus variable-number tandem-repeat analysis (MLVA).

Listeria monocytogenes is responsible for severe and often fatal food-borne infections in humans. A collection of 2,421 L. monocytogenes isolates originating from Ontario's food chain between 1993 and 2010, along with Ontario clinical isolates collected from 2004 to 2010, was characterized using an improved multilocus variable-number tandem-repeat analysis (MLVA). The MLVA method was established based on eight primer pairs targeting seven variable-number tandem-repeat (VNTR) loci in two 4-plex fluorescent PCRs. Diversity indices and amplification rates of the individual VNTR loci ranged from 0.38 to 0.92 and from 0.64 to 0.99, respectively. MLVA types and pulsed-field gel electrophoresis (PFGE) patterns were compared using Comparative Partitions analysis involving 336 clinical and 99 food and environmental isolates. The analysis yielded Simpson's diversity index values of 0.998 and 0.992 for MLVA and PFGE, respectively, and adjusted Wallace coefficients of 0.318 when MLVA was used as a primary subtyping method and 0.088 when PFGE was a primary typing method. Statistical data analysis using BioNumerics allowed for identification of at least 8 predominant and persistent L. monocytogenes MLVA types in Ontario's food chain. The MLVA method correctly clustered epidemiologically related outbreak strains and separated unrelated strains in a subset analysis. An MLVA database was established for the 2,421 L. monocytogenes isolates, which allows for comparison of data among historical and new isolates of different sources. The subtyping method coupled with the MLVA database will help in effective monitoring/prevention approaches to identify environmental contamination by pathogenic strains of L. monocytogenes and investigation of outbreaks.

Pandemic H1N1 Influenza Infection and Vaccination in Humans Induces Cross-Protective Antibodies that Target the Hemagglutinin Stem.

Most monoclonal antibodies (mAbs) generated from humans infected or vaccinated with the 2009 pandemic H1N1 (pdmH1N1) influenza virus targeted the hemagglutinin (HA) stem. These anti-HA stem mAbs mostly used IGHV1-69 and bound readily to epitopes on the conventional seasonal influenza and pdmH1N1 vaccines. The anti-HA stem mAbs neutralized pdmH1N1, seasonal influenza H1N1 and avian H5N1 influenza viruses by inhibiting HA-mediated fusion of membranes and protected against and treated heterologous lethal infections in mice with H5N1 influenza virus. This demonstrated that therapeutic mAbs could be generated a few months after the new virus emerged. Human immunization with the pdmH1N1 vaccine induced circulating antibodies that when passively transferred, protected mice from lethal, heterologous H5N1 influenza infections. We observed that the dominant heterosubtypic antibody response against the HA stem correlated with the relative absence of memory B cells against the HA head of pdmH1N1, thus enabling the rare heterosubtypic memory B cells induced by seasonal influenza and specific for conserved sites on the HA stem to compete for T-cell help. These results support the notion that broadly protective antibodies against influenza would be induced by successive vaccination with conventional influenza vaccines based on subtypes of HA in viruses not circulating in humans.

Tasco supplementation: effects on carcass characteristics, sensory attributes, and retail display shelf-life.

Two hundred crossbred cattle (Bos indicus x Bos taurus) were supplemented with 2% Tasco (Ascophyllum nodosum) in a commercial finishing facility to evaluate marbling score, USDA quality grade, sensory traits, and retail display shelf life. Treatment animals (n = 100) received a steam-rolled corn (Zea mays)-based diet containing 2% Tasco meal (DM basis), for 14 d beginning at d 45 of the finishing period and again 14 d before slaughter. Control animals (n = 100) received a steam-rolled corn (Zea mays)-based diet without Tasco at identical feeding periods. Carcasses from Tasco-fed cattle exhibited greater marbling scores (P = 0.003) than controls. There were no treatment effects (P > 0.05) on sensory, shear, or purge attributes of striploin or inside round steaks with the exception of inside round steaks from Tasco animals having a greater initial tenderness (P = 0.03) and lower off-flavor score (P = 0.002) than control steaks. The LM samples from Tasco-fed cattle had a greater percentage of ether extractable fat (P = 0.001) and lower percentage of protein (P = 0.001) than controls. Inside round samples from treated animals exhibited a greater percentage of moisture (P = 0.03) than control steaks. Visual lean color of striploin steaks was not affected by Tasco supplementation (P = 0.26); however, steaks from Tasco-treated animals were more uniform and had less discoloration and browning than those from controls (P = 0.005, 0.04, and 0.05, respectively). Inside round visual scores and instrument values reflected similar treatment responses (P < 0.05), with a majority of the effects on muscle redness (CIE a*, hue angle) and measures of discoloration. Tasco steaks were generally more red and less discolored during extended postmortem aging and retail exposure (P < 0.05). The results from this study indicate that short-term supplementation of 2% Tasco meal in feedlot cattle increases carcass quality and prolonged retail shelf life.

Behavior, preference for, and use of alfalfa, tall fescue, white clover, and buffalograss by pregnant gilts in an outdoor production system.

Sustainable outdoor pig production requires vegetation that can maintain ground cover, assimilate manure nutrients, and prevent soil erosion. Two experiments were conducted to evaluate the suitability of four forages: alfalfa (Medicago sativa), tall fescue (Festuca arundinacea), white clover (Trifolium repens), and buffalograss (Buchloe dactyloides) for grazing or ground cover in pastures for pigs. Each forage plot covered 7.5 m2, with nine replicates in a randomized block design. In Exp. 1, eight pregnant gilts had free access to all forages during a 2-d adjustment period. Immediately thereafter, pairs of gilts were assigned randomly to one of each of four blocks of the four forages during a 2-d measurement period. The percentage of ground cover for each forage was visually estimated at 0, 24, and 48 h of study. Behavioral data, including walking, eating, grazing, rooting, drinking, standing, lying, and time spent in hut were video-monitored continuously for 48 h. Initial percentage of ground cover was 100% for all species. By 48 h, percentage of ground cover decreased (P < 0.001) for white clover (11.3 +/- 0.88%) and alfalfa (36.3 +/- 0.88%), but not for tall fescue (98.0 +/- 0.88%) or buffalograss (98.0 +/- 0.88%). Gilts spent more (P < 0.01) time grazing white clover (16.3 +/- 1.97 min/d) and alfalfa (11.2 +/- 1.97) than tall fescue (0.8 +/- 1.97) or buffalograss (0.3 +/- 1.97), and rooted more (P < 0.04) white clover than other forages. In Exp. 2, six gilts from the initial group were put on six blocks of the four forages. Each gilt was assigned randomly to three replicates of each forage, including alfalfa, tall fescue, or buffalograss (white clover was excluded because of damage by gilts during Exp. 1), and gilts grazed single forages for 2 d. After this grazing period, the percentage of ground cover was less (P < 0.01) for alfalfa than for buffalograss or tall fescue (37.5 +/- 0.38, 96.7 +/- 0.39, 96.3 +/- 0.39%, respectively). With access to a single forage, pregnant gilts spent more (P < 0.01) time grazing alfalfa (15.8 +/- 2.36 min/d) than buffalograss (1.5 +/- 2.36) or tall fescue (0.7 +/- 2.37). These gilts clearly preferred grazing white clover and alfalfa, and rooting and eating white clover compared with buffalograss or tall fescue. Rates of ground cover loss were less (P < 0.01) for tall fescue and buffalograss than for the more preferred forages. Less preferred forages could have potential as pasture for swine when the primary objective is ground cover maintenance rather than nutrient supply.

Effect of supplemental vitamin D3 concentration on concentrations of calcium, phosphorus, and magnesium relative to protein in subcellular components of the longissimus and the distribution of calcium within longissimus muscle of beef steers.

The effect of supplementing diets with various levels of vitamin D3 to provide 0, 0.5, 1, and 5 million IU/(steer x d) for 8 d before slaughter on the mineral content and localization of Ca in LM and muscle fragments was studied during the postmortem aging process. Twelve feedlot steers of three biological types were given access to the four levels of vitamin D for 8 d before slaughter. Differential centrifugation techniques were used to determine the concentrations of minerals relative to protein in different muscle fragments on d 3 and 21 postmortem. Electron microscopy visualization of bound Ca indicated that vitamin D3 mobilized Ca from the sarcoplasmic reticulum and transverse tubule system into the myofibrils. Bound Ca was concentrated near the Z-line at the A-band/I-band juncture within the sarcomere. Supplementing steers with 1 and 5 million IU/(steer x d) of vitamin D3 increased (P < 0.05) Ca, P, and Mg concentrations per unit of protein in the cytosol. Soluble cytosolic Ca concentrations were greater (P < 0.05) on d 21 than on d 3 postmortem only when steers were supplemented with 5 million IU/d. Concentrations of Ca, P, and Mg in isolated tissues were increased (P < 0.05) in nuclei and myofibrilar proteins by supplementing steers with 1 and 5 million IU/ (steer x d) of vitamin D3. All supplemental vitamin D3 treatments also increased (P < 0.001) Mg concentrations in the cytosol, regardless of aging treatment, and increased Mg concentrations (P < 0.04) within the mitochondria at d 3 postmortem. Thus, supplementation of feedlot steers with vitamin D3 at levels of 0.5 to 5 million IU/(steer x d) increased Ca concentrations within respiring muscle, resulting in increased bound tissue Ca concentrations. When the respiring muscle was converted to meat, the increased bound tissue Ca resulting from vitamin D3 treatment released Ca concentrations into the cytosol during aging (P < 0.05). Results of this study indicate that vitamin D3 supplementation increased total cytosolic Ca, P, and Mg concentrations in meat.

Ascophyllum nodosum supplementation: a preharvest intervention for reducing Escherichia coli O157:H7 and Salmonella spp. in feedlot steers.

Ascophyllum nodosum (Tasco-14) decreased the prevalence of enterohemorrhagic Escherichia coli (EHEC) O157:H7 in animals fed prior to harvest. Tasco-14 was supplemented on a 2% dry matter basis 14 days prior to harvest to determine its effects on EHEC and Salmonella spp. prevalence. Two hundred mixed crossbred steers and heifers (Bos indicus x Bos taurus), in a large commercial finishing facility, served as experimental units. Treatment (TRT, n = 100) animals received a steam-rolled corn-based diet containing 2% Tasco-14 on a dry matter basis, and control (CON, n = 100) animals received only the steam-rolled corn-based diet. Hide swabs and fecal samples were obtained for EHEC and Salmonella spp. evaluations. Animals were sampled 1 day prior to (d - 1) the feeding of Tasco-14 and immediately following exsanguinations. The prevalence of EHEC O157 on hide swabs and in fecal samples (P = 0.0001 and P < 0.0001, respectively) and the prevalence of EHEC O157:H7 on hide swabs and in fecal samples (P < 0.0001 and P < 0.0001, respectively) was reduced by 33 and 36% from d - 1 levels on TRT hide swabs and by 9 and 11% in TRT fecal samples. The prevalence of EHEC O157 and EHEC O157:H7 was reduced by 33 and 36% from d - 1 levels on TRT hide swabs and by 9 and 11% in TRT fecal samples. The prevalence of Salmonella spp. on hide swabs did not change for TRT animals (P = 0.64). CON animals showed an increase in Salmonella spp. prevalence (P < 0.0001) from d - 1 feeding levels on hide swabs. The prevalence of Salmonella spp. increased in both TRT and CON fecal samples when compared to d - 1 levels (P = 0.002). However, TRT samples exhibited a lower (P < 0.05) postfeeding prevalence of Salmonella spp. in fecal samples than did CON samples. Results from this study indicate that 2% Tasco-14 supplementation in feedlot cattle diets reduces EHEC O157 and EHEC O157:H7 prevalence on hide swabs and in fecal samples and may suppress increases in Salmonella spp.

Matua bromegrass hay for mares in gestation and lactation.

Matua bromegrass hay (Bromus willdenowii Kunth) is a high quality forage, but its value for mares during gestation and lactation is not well known. Intake, rate of passage, performance, and reproduction by gestating and lactating Quarter Horse mares fed the hay was investigated. In this experiment, 12, 2- to 12-yr-old gravid mares (mean BW = 553 kg, SD = 36) were fed Matua hay (CP = 11.5%) or alfalfa hay (Medicago sativa L.) (CP = 15.4%) for variable days prepartum (mean 59.9 d; SD = 23.5) and for 70 d postpartum. Matua and alfalfa hay were fed as the roughage portion of the diet with a grain supplement. Mares, blocked by age, expected date of foaling, and BW, were assigned randomly within blocks to treatments (six mares per treatment). Forage type did not affect intake, gestation length, birth weight, number of foals, foal weight gain, day of first postpartum ovulation, cycles per conception, or pregnancy rate at 70 d. On d 1, milk from mares fed alfalfa hay contained less (P < 0.03) CP than milk from mares fed Matua hay. Milk CP decreased (P < 0.01) in all mares over time. In a separate experiment, voluntary intake and rate of passage of Matua (CP = 15.5%), alfalfa (CP = 24.9%), and Timothy (Phleum pratense L.) (CP = 4.1%) hays were determined in nine 2-yr-old pregnant mares (mean BW = 447 kg; SD = 21). Diets were 100% forage. Timothy hay did not meet CP requirements for mares. Voluntary intake of alfalfa hay was higher (P < 0.01) than Matua hay. Intake of Timothy hay was lower (P < 0.01) than the mean of alfalfa and Matua hay. Rate of passage offorage was measured by passage of Cr-mordanted fiber. Passage rate and retention time did not differ between Matua and alfalfa hay; however, the retention times of Matua and alfalfa hays were shorter (P < 0.01) than for Timothy hay. Our results indicate that Matua hay is a forage that can be used safely for mares during gestation and early lactation and for their young foals.

Tasco-Forage: I. Influence of a seaweed extract on antioxidant activity in tall fescue and in ruminants.

Seaweed (Ascophyllum nodosum) is a known source of plant growth regulators, and application to turfgrasses has increased activity of the antioxidant superoxide dismutase (SOD) and specific vitamin precursors. Increased antioxidant activity in both plants and animals diminishes oxidative stress. Two pasture experiments investigated effects of Tasco-Forage (a proprietary seaweed-based product) applied to tall fescue (Festuca arundinacea) on antioxidant activity in plants and in ruminants that grazed the forage. In Exp. 1, fescue was 70 to 100% infected with the endophyte fungus Neotyphodium coenophialum ([Morgan-Jones and Gams] Glenn, Bacon, and Hanlin). Twenty-four wether lambs (initial BW 41 kg; SD = 5) grazed fescue treated with 0, 1.7, or 3.4 kg Tasco/ha applied in April and July, 1994, with four replications per treatment. Grazing occurred for 26 d beginning April 21 and for 22 d beginning July 19. In July, there was a linear increase in daily gains (P < 0.05), and serum vitamin A (P < 0.13) and whole-blood Se (P < 0.10) tended to increase in lambs grazing Tasco-treated fescue. In Exp. 2,48 Angus and Angus x Hereford steers (initial BW 245 kg; SD = 20) grazed infected or uninfected tall fescue in Virginia that was treated (3.4 kg/ ha) or untreated with Tasco in April and July, 1995. Steers that grazed infected tall fescue had lower (P < 0.02) serum vitamin A and E and tended (P < 0.07) to have lower whole-blood Se in September compared with steers that grazed uninfected tall fescue. Tasco decreased (P < 0.06) serum vitamin E but tended to increase whole-blood Se (P < 0.10) in September and serum vitamin A in July (P < 0.12). During 1996 and 1997, the experiment was repeated in Virginia with Angus steers and was replicated at Prairie, MS, where 1/4 Brahman x 3/4 Angus steers were used. Forty-eight steers were included at each location in each year (n = 192 total steers for 1996 and 1997). Steers that grazed infected tall fescue in Mississippi had lower (P < 0.05) serum vitamin E by the end of the grazing season. At both locations Tasco increased (P < 0.05) activity of superoxide dismutase in both infected and uninfected fescue. The endophyte in tall fescue seemed to decrease antioxidant activity in grazing steers, whereas Tasco seemed to increase antioxidant activity in both the forage and the grazing ruminant. Tasco may provide opportunities to reduce oxidative stress in plants and animals.

Tasco-Forage: II. Monocyte immune cell response and performance of beef steers grazing tall fescue treated with a seaweed extract.

Effects of applying Tasco-Forage, an Ascophyllum nodosum seaweed-based product prepared by a proprietary process, to endophyte (Neotyphodium coenophialum [Morgan-Jones and Gams] Glenn, Bacon, and Hanlin)-infected and endophyte-free tall fescue (Festuca arundinacea Schreb.) were studied in each of 3 yr (1995, 1996, and 1997) in Virginia and in 1996 and 1997 in Mississippi. There were 48 steers at each location in each year (n = 240) in a 2 x 2 x 2 factorial arrangement with two replications at each location. Steers in Virginia were Angus and Angus x Hereford with initial weights of 245 kg (SD = 20), 234 kg (SD = 9), and 265 kg (SD = 5) in yr 1, 2, and 3, respectively. Steers in Mississippi were 3/4 Angus and 1/4 Brahman and weighed 230 kg (SD = 8) and 250 kg (SD = 2) in yr 2 and 3, respectively. Tasco (3.4 kg/ha) was dissolved in water and applied to pastures in April before grazing was begun and again in July at the same rate. The grazing period was from mid-April to late September or mid-October. Total gains were higher (P < 0.05) for steers grazing uninfected than for those grazing endophyte-infected tall fescue. Rectal temperatures were increased (P < 0.05) due to endophyte infection at both locations; Tasco application decreased temperature of steers grazing infected fescue in Virginia (interaction, P < 0.07) but increased temperatures of steers grazing infected fescue in Mississippi (interaction, P < 0.05). Presence of the endophyte resulted in rough hair coats and loss of hair color, but the effect was partially offset (P < 0.05) by Tasco application in Virginia in 1995. Both monocyte phagocytic activity (all years and locations) and major histocompatibility complex class II expression (1995 only) were decreased (P < 0.05) in steers due to endophyte infection, but this effect was reversed (P < 0.05) by application of Tasco to pastures. Application of the extract from A. nodosum seems to have use in alleviating adverse effects of endophyte on immune function and may improve hair coat condition in cattle grazing infected fescue, but effects on rectal temperature varied due to location.

Tasco-Forage: III. Influence of a seaweed extract on performance, monocyte immune cell response, and carcass characteristics in feedlot-finished steers.

Tall fescue (Festuca arundinacea Schreb.) infected with the endophyte Neotyphodium coenophialum ([Morgan-Jones and Gams] Glenn, Bacon, and Hanlin) causes fescue toxicosis in cattle grazing the forage, but effects of the endophyte were considered to be abated soon after removal of the animals from pastures. Tasco-Forage, a proprietary extract from the brown seaweed Ascophyllum nodosum, is a known source of cytokinins and has increased antioxidant activity in both plants and the animals that graze the forage. Tasco was applied at 0 and 3.4 kg/ha to infected and uninfected tall fescue pastures in Virginia and Mississippi. Forty-eight steers grazed the pastures at each location during each of 2 yr (n = 192) before being transported to Texas for feedlot finishing. On arrival at the feedlot, steers from Tasco-treated pastures had higher (P < 0.01) monocyte phagocytic activity and tended (P < 0.07) to have higher major histocompatibility complex class II expression than steers that grazed the untreated pastures. A depression (P < 0.05) in monocyte immune cell function due to grazing infected fescue was detected throughout the feedlot finishing period but was reversed by Tasco. Rectal temperatures were elevated (P < 0.07) in steers that had grazed the infected tall fescue when they arrived in Texas, but by d 14 no difference was detected. However, by d 28 the temperature effects of infected tall fescue were reversed. Steers that had grazed infected fescue had lower (P < 0.01) rectal temperatures on d 112 of the feedlot period, demonstrating a much longer-lasting effect of the endophyte on thermoregulatory mechanisms than previously thought. Steers that had grazed Tasco-treated pastures had higher (P < 0.01) rectal temperatures on d 56 than steers that had grazed untreated fescue. Steers that had grazed the Tasco-treated pastures had higher marbling scores (P < 0.05) regardless of the endophyte, but no effect of Tasco or endophyte on gain was measured. Our data suggest that Tasco application to tall fescue pastures alleviated some of the negative effects of tall fescue toxicity.

Tasco-Forage: IV. Influence of a seaweed extract applied to tall fescue pastures on sensory characteristics, shelf-life, and vitamin E status in feedlot-finished steers.

Tasco-Forage is an Ascophyllum nodosum seaweed-based product that has increased antioxidant activity in both plants and animals. Endophyte (Neotyphodium coenophialum ([Morgan-Jones and Gams] Glenn, Bacon, and Hanlin)-infected and uninfected tall fescue (Festuca arundinacea Schreb.) pastures in Virginia and Mississippi during 1997 were treated or not with 3.4 kg Tasco/ha in April and July. There were two replications of each treatment at each location. Forty-eight steers (6/replication) grazed pastures at each location (n = 96) from April to October prior to transportation to Texas Tech, Lubbock, for finishing during a 160-d period in the feedlot. Blood (antemortem) and liver (postmortem) samples were collected. After slaughter and chilling, the left strip loins (IMPS #180) were collected from three randomly selected steers from within each pasture replication (n = 48). Strip loins were vacuum-packaged and stored at 2 degrees C. At postmortem d 7, 14, 21, and 28, strip loins were removed from packaging and fabricated into 2.54-cm steaks. Following each fabrication day postmortem, the strip loins were repackaged and stored at 2 degrees C until the following postmortem time. After the prescribed fabrication, steaks were overwrapped with polyvinyl chloride film, subjected to simulated retail display at 2 degrees C for up to 3 d, and subjective and objective color were evaluated daily by a trained panel. Steaks from Mississippi steers that had grazed Tasco-treated fescue retained higher (P < 0.05) CIE a* color scores throughout retail display. Steaks were more uniform and had less discoloration and less browning (P < 0.05) if they were from steers that had grazed Tasco-treated fescue, and the effect was greatest for steers from Mississippi (location x Tasco interaction; P < 0.05). The endophyte in tall fescue may decrease uniformity and increase lean discoloration and two-toning of beef steaks when removed from vacuum packaging on or beyond d 21 postmortem (endophyte x Tasco x postmortem day interaction: P < 0.05). Vitamin E in liver was increased (P < 0.06) and serum vitamin E was decreased (P < 0.09) in steers that had grazed the treated pastures. These experiments indicated that Tasco applied to tall fescue during the grazing season can improve color stability and extend beef shelf-life, particularly in cattle grazing infected tall fescue. The mode of action of Tasco is not clear, but antioxidants and specific vitamins may be involved.

Forage systems for production of stocker steers in the upper south.

The southern states produce large numbers of beef calves that are generally weaned and sold in autumn. Keeping calves in this region beyond weaning to graze high-quality forages through a stocker cattle phase could improve profitability. Autumn-weaned Angus crossbred steers were allocated by breeding and weight to four forage systems that began in mid-November and continued through mid-October as follows: System 1, tall fescue (Festuca arundinacea Schreb.) and Kentucky bluegrass (Poa pratensis L.)-white clover (Trifolium repens L.); System 2, tall fescue, caucasian bluestem (Bothriochloa caucasica [Trin.] C. E. Hubbard) and tall fescue-red clover (Trifolium pratense L.); System 3, orchardgrass-alfalfa and bluegrass-white clover; and System 4, rye (Secale cereale L.), soybeans (Glycine max)-foxtail millet (Setaria italica), and bluegrass-white clover. All steers were supplemented with hay or silage previously cut from their respective systems when forage for grazing was limited. System 2 which used stockpiled tall fescue for winter grazing and caucasian bluestem for summer forage plus fescuered clover for hay and grazing in a three-paddock system, resulted in greater (P < .01) gain per hectare and per steer, more grazing days, and reduced stored forage requirements and produced more surplus feed than the other systems tested. Gains per hectare for Systems 1 through 4 were 454, 554, 472, and 487 kg (SE = 18), respectively. Harvested forage from Systems 1, 2, and 3 met needs for stored forages but System 4 required additional "purchased" hay. Stored forage was fed for 61, 38, 112, or 104 d for Systems 1 through 4, respectively. Within the physio-climatic region of this experiment, a simple three-paddock system based on cool- and warm-season perennial forages could improve beef production per unit of land area while reducing inputs of labor and equipment.

Voluntary intake, digestibility, and subsequent selection of Matua bromegrass, coastal bermudagrass, and alfalfa hays by yearling horses.

Matua bromegrass (Bromus willdenowii Kunth. cv. Grasslands Matua) was introduced in 1973, but little information exists concerning its potential as a hay for horses. Thus, voluntary intake and apparent digestibility of OM, CP, and fiber components of Matua by 18 Quarter Horse yearlings (mean initial BW 354 kg; SE 5.8) were compared with alfalfa (Medicago sativa L.) and coastal bermudagrass (Cynodon dactylon L.) as hays in a randomized block design. A 15-d adjustment period was followed by a 5-d collection period during which the hays were consumed ad libitum. Voluntary intake of DM was greater (P<.01) for alfalfa (10.9 kg/d) than for the mean of the grasses, and intake of Matua (10.0 kg/d) was greater (P<.001) than that of bermudagrass (7.4 kg/d). Apparent digestibility of OM was greater (P<.001) for alfalfa (74%) than for the mean of the grasses but did not differ between Matua (64%) and bermudagrass (60%). At the end of the digestion trial, each yearling was offered each of the three forage hays during an 11-d period to determine subsequent preference and effect of previous hay experience. Yearlings preferred alfalfa over the grass hays and generally selected more Matua than bermudagrass. All yearlings consumed less of the forage species to which they had been previously exposed compared with unadapted yearlings. The Matua hay fed in this trial was palatable and met most of the nutritional needs for yearling horses.

Toward a framework for computer-mediated collaborative design in medical informatics.

The development and implementation of enabling tools and methods that provide ready access to knowledge and information are among the central goals of medical informatics. The need for multi-institutional collaboration in the development of such tools and methods is increasingly being recognized. Collaboration involves communication, which typically involves individuals who work together at the same location. With the evolution of electronic modalities for communication, we seek to understand the role that such technologies can play in supporting collaboration, especially when the participants are geographically separated. Using the InterMed Collaboratory as a subject of study, we have analyzed their activities as an exercise in computer- and network-mediated collaborative design. We report on the cognitive, sociocultural, and logistical issues encountered when scientists from diverse organizations and backgrounds use communications technologies while designing and implementing shared products. Results demonstrate that it is important to match carefully the content with the mode of communication, identifying, for example, suitable uses of E-mail, conference calls, and face-to-face meetings. The special role of leaders in guiding and facilitating the group activities can also be seen, regardless of the communication setting in which the interactions occur. Most important is the proper use of technology to support the evolution of a shared vision of group goals and methods, an element that is clearly necessary before successful collaborative designs can proceed.

Influence of Neotyphodium coenophialum on copper concentration in tall fescue.

Poor performance of livestock that graze tall fescue (Festuca arundinacea Schreb.) has been associated with the endophyte fungus Neotyphodium coenophialum [Morgan-Jones and Gams] Glenn, Bacon, and Hanlin). Recent evidence suggests lowered Cu status and a depression of Cu-related immune function in steers that graze endophyte-infected (E+) tall fescue. Greenhouse and field studies investigated relationships between the endophyte and Cu concentrations in tall fescue. Seventeen infected 'Kenhy' clones were divided, and one plant of each pair was treated three times with Benomyl to remove the endophyte (E-). Plants were watered with nutrient solution in a greenhouse for 6 mo before sampling. Copper concentrations were greater (P < .001) in E- than in E+ clones (3.4 vs 2.8 microg/g; SE, .06). In the second greenhouse experiment, genetically similar E+ and E- 'Kentucky'-31 (KY-31) and 'Georgia Jessup' were grown from seed and fertilized with nutrient solution to produce mature plants. Copper concentrations were higher (P < .05) in E- than in E+ tall fescue (8.6 vs 7.6 microg/g; SE, .3). In a field plot experiment in Texas, E+ and E- KY-31 were grown with 0, 50, and 100% replacement of potential evapotranspiration. By September, Cu concentrations were higher (P < .05) in E- than in E+ tall fescue (7.3 vs 6.6 microg/g; SE, .2). In pasture experiments, KY-31 E+ (> 70% infection level) and E- (< 5% infection level) tall fescue were grown in Virginia at two locations with three rates of N fertilizer. Copper concentrations were higher (P < .05) in E- than in E+ tall fescue (4.8 vs 4.5 microg/g; SE, .1) and increased (P < .01) linearly in response to N. Our data demonstrate that the presence of the endophyte is associated with lower Cu concentrations in tall fescue, which may contribute to lowered Cu status in animals and thus contribute to the etiology of fescue toxicity.

Monocyte immune cell response and copper status in beef steers that grazed endophyte-infected tall fescue.

A 3-yr study was conducted to evaluate immune response and Cu status of yearling beef steers as a consequence of grazing tall fescue (Festuca arundinacea Schreb.) infected (E+) with the endophyte fungus Neotyphodium coenophialum ([Morgan-Jones and Gams] Glenn, Bacon, and Hanlin). During a preliminary study in 1994, 24 weanling Angus and Angus x Hereford steers were blocked by breed and weight (initial BW 271 kg; SD 25) and were randomized to E+ and low endophyte (E-) fescue in pastures at Glade Spring, VA. Grazing began in April and was discontinued in July. In 1995 and 1996, 24 weanling Angus and Angus x Hereford steers (initial BW 249 kg, SD 20 and 240 kg, SD 15, respectively) were randomized to the E+ and E- pastures at Glade Spring during each year. Grazing began in April and continued until September in 1995 and October in 1996. In 1994, steers that grazed E+ fescue exhibited lower (P < .05) phagocytic activity, major histocompatibility complex (MHC) class II expression, ceruloplasmin, and serum Cu than steers that grazed E- tall fescue. During 1995, steers grazing E+ fescue had lower (P < .05) phagocytic activity and MHC class II expression than steers that grazed E- fescue. In 1996, one-half of the steers within each paddock received a Cu oxide bolus at the beginning of the grazing season. During 1996, phagocytic activity was lower (P < .01) and MHC class II expression tended (P < .07) to be lower in steers that grazed E+ tall fescue than in steers that grazed E- tall fescue. Copper supplementation increased (P < .05) MHC class II expression in July regardless of endophyte status over nonsupplemented steers. Steers that grazed E- tall fescue had higher (P < .05) plasma or serum Cu concentrations than steers that grazed E+ tall fescue in each year of the study. These data indicate that the endophyte compromised the immune function of grazing steers, and the data suggest a relationship with depressed Cu status.

Evaluating evidence against diagnostic hypotheses in clinical decision making by students, residents and physicians.

Clinicians are faced with the difficulty of filtering large quantities of information and incorporating evidence to make safe and accurate diagnostic, therapeutic and management decisions. One solution to this difficulty is the development of evidence-based decision support tools designed to provide relevant and up-to-date evidence to clinicians. However, as investigations of medical decision making have found that hypothesis generation and clinical reasoning differ as a function of expertise, the gathering, interpretation and use of evidence against hypotheses depends on the prior knowledge of the clinician. The specific focus of the study is to understand how clinical evidence is gathered and evaluated during diagnostic reasoning. Verbal data during clinical interactions with physicians, residents and final year medical students were collected and transcribed. The dialogues were analyzed using three levels of coding: cognitive, epistemological and linguistic pragmatic perspectives. Results show that: (a) the ability to index and use adequate evidence by physicians, residents and students is a function of the early generation of accurate hypotheses: and (b) strategies for resolving inconsistent evidence differ as a function of medical expertise. The relationship between these findings and its application for the development of adequate knowledge-based systems for indexing and retrieval is discussed.