Recirculating elutriator for extracting gastrointestinal nematode larvae from pasture herbage samples.

Gastrointestinal nematode (GIN) parasites present an important limitation to ruminant production worldwide. Methods for quantifying infective larvae of GIN on pastures are generally tedious, time-consuming, and require bulky equipment set-ups. This limitation to expedient data collection is a bottleneck in development of pasture management practices that might reduce pasture infectivity. We modified a soil elutriator concept for extracting GIN larvae from fresh herbage samples. Elutriators were constructed from readily available parts and compared to the Baermann funnel sedimentation method for larvae extraction. More samples could be extracted per day in the elutriator than in a Baermann unit with extraction times of 8 min versus 24h, respectively. Accuracy, measured as maximum recovery of larvae seeded onto herbage samples, did not differ between extraction methods (62.3 vs. 69.8% for elutriator and Baermann, respectively, P>0.05). Larvae recovery from herbage in elutriators showed a strong log(e) relationship with extraction time (r(2)>0.98), which will allow development of accurate correction factors for specific herbages to predict total larvae densities at extraction times less than those needed for maximum recovery. An extraction time of 8 min per sample gave the best compromise of speed, accuracy, and precision as measured by regression confidence bands and root mean square error of analysis of variance. Precision of the elutriator extraction for pasture samples was comparable to published methods and was not affected by forage species or canopy strata. The elutriator method was sensitive enough to detect differences in larvae density as small as 8 larvae g(-1) DM among pasture treatments. Elutriators extracted nematode larvae from herbage samples with accuracy and precision similar to existing methods, but did it much faster. Elutriation shows promise as a rapid method for extracting infective GIN larvae from pasture herbage.

Alpha-tocopherol concentrations and case life of lamb muscle as influenced by concentrate or pasture finishing.

Two experiments were conducted to evaluate alpha-tocopherol accumulation in muscle of lambs finished on pasture or concentrates. The objective for Exp. 1 was to compare accumulation of alpha-tocopherol in the longissimus muscle of pasture-fed lambs to that of lambs fed three concentrations (15, 150, and 300 IU/kg of DM) of supplemental vitamin E (all rac alpha-tocopheryl acetate) in all-concentrate diets. The objective in Exp. 2 was to investigate the effect of duration of supplemental vitamin E feeding on alpha-tocopherol content and color change during display case storage of lamb muscle. Treatments evaluated in Exp. 2 were: 15 IU of supplemental vitamin E/kg DM fed to finish; 15 IU/kg followed by 300 IU/kg of DM during the last 21 d; and 15 IU/kg DM until 7 d prior to finish, then 300 IU/kg DM. In Exp. 1, alpha-tocopherol concentration of rotational grazed alfalfa and perennial ryegrass averaged 137 and 169 mg/kg of DM. Vitamin E treatments for lambs fed concentrate diets did not affect ADG (P > 0.15), but ADG was greater (P < 0.01) for concentrate-fed lambs than for grazing lambs. For the concentrate-fed lambs, alpha-tocopherol in longissimus muscle increased quadratically (P < 0.05) as dietary concentrations of vitamin E increased. Predicted maximum alpha-tocopherol concentration in muscle occurred at about 400 IU/kg of diet DM. Longissimus muscle from lambs grazing alfalfa or ryegrass had similar (P > 0.50) alpha-tocopherol concentrations, and those concentrations were similar to values obtained when the concentrate diet supplemented with 150 IU of vitamin E/kg was fed. In Exp. 2, no differences (P > 0.10) in ADG were observed. Concentrations of longissimus alpha-tocopherol were highest when 300 IU supplemental vitamin E was fed for 21 d prior to slaughter. During a 6-d display period, semimembranosus steaks from lambs fed 300 IU of supplemental vitamin E/kg for either 7 or 21 d had higher a* and b* color readings than steaks from lambs fed 15 IU/kg of supplemental vitamin E. Increased consumption of vitamin E either via pasture or supplementation results in higher alpha-tocopherol concentrations in meat.

A national survey of critical care nurses' practices related to administration of neuromuscular blocking agents.

BACKGROUND: Recommendations on use of neuromuscular blocking agents include using peripheral nerve stimulators to monitor depth of blockade and concomitantly administering sedatives and/or analgesics. OBJECTIVE: To evaluate critical care nurses' practices in administering neuromuscular blocking agents. METHODS: A 16-item survey was mailed to 483 acute care facilities in the United States. Of these, 246 surveys (51%) were returned and analyzed to determine use of neuromuscular blocking agents, peripheral nerve stimulators, sedatives, and analgesics. Logistic regression analysis was used to find independent predictors of use of peripheral nerve stimulators. RESULTS: Seventy-five percent of respondents reported long-term use of neuromuscular blocking agents in critically ill patients. Of those, 63% monitored the level of blockade with peripheral nerve stimulators. Reasons for not using peripheral nerve stimulators included unavailability of equipment (48%), lack of training (36%), and insufficient evidence that peripheral nerve stimulators improve care (23%). Predictors of use of stimulators were facilities with more than 150 beds (P < .001) and administration of neuromuscular blocking agents by continuous infusion (P < .001). Ninety-five percent of respondents reported using concurrent analgesics/sedatives always or most of the time. Facilities with fewer than 10 beds in the intensive care unit used concurrent analgesics/sedatives significantly less often than did facilities with 10 beds or more (90% vs 98%, respectively; P = .03). CONCLUSIONS: Small and large facilities differ in concomitant use of analgesics/sedatives and peripheral nerve stimulators. Education and research are needed to ensure that patients receive adequate monitoring and sedation during administration of neuromuscular blocking agents.

Influence of glyphosate on amino acid composition of Egyptian broomrape.

The parasitic plant broomrape is entirely dependent on its host for reduced carbon and nitrogen and is also susceptible to inhibition by glyphosate that is translocated to the parasite through a host. Studies were conducted to examine the effect of broomrape parasitism on amino acid concentrations of two hosts: common vetch that is tolerant of low levels of glyphosate and oilseed rape that has been genetically engineered for glyphosate resistance. The influence of glyphosate on the amino acid content of broomrape and the two hosts was also examined. Amino acid concentrations in leaves and roots of parasitized common vetch plants were generally similar to those of the corresponding tissues of nonparasitized plants. Amino acid concentrations in broomrape were lower than those of the parasitized common vetch root. For common vetch, glyphosate applied at rates that selectively inhibited broomrape growth did not alter individual amino acid concentrations in the leaves, but generally increased amino acid levels at 0.18 kg ha-1. Glyphosate application also increased the amino acid concentrations, with the exception of arginine, of broomrape growing on common vetch and did not generally influence concentrations in leaves or roots of common vetch. In oilseed rape, parasitization by broomrape generally led to higher amino acid concentrations in leaves but lower concentrations in roots of parasitized plants. Broomrape had higher amino acid concentrations than roots of the parasitized oilseed rape. Glyphosate applied at 0.25 and 0.5 kg ha-1 generally increased the amino acid concentrations in oilseed rape leaves, but the 0.75 kg ha-1 application caused the amino acid concentrations to decrease compared to those of untreated plants. In oilseed rape root the general trend was an increase in the concentration of amino acids at the two highest rates of glyphosate. Individual amino acid concentrations in broomrape attachments growing on oilseed rape were generally increased following glyphosate application of 0.25 kg ha-1. These results indicate that low rates of glyphosate alter amino acid profiles in both host and broomrape and raise questions about the regulation of amino acid metabolism in the parasite.

Impact of egyptian broomrape (Orobanche aegyptiaca (Pers.) parasitism on amino acid composition of carrot (Daucus carota L.).

The relationship between the organic nitrogen status of Egyptian broomrape and one of its hosts, carrot, was studied by comparing amino acid profiles of leaf and root tissues of nonparasitized and broomrape-parasitized carrot plants and by analyzing amino acid profiles of broomrape at different growth stages. Total N concentrations, expressed as a percentage of the dry weight of the tissues, were similar in leaves of nonparasitized and parasitized carrot plants but were lower in parasitized roots than in nonparasitized roots. In both dry and germinated broomrape seeds, N concentrations were lower than or similar to those in broomrape tubercles, shoots, or callus. Individual amino acid concentrations in hydrolysates of leaves of parasitized carrot plants tended to be similar to or greater than those in hydrolysates of nonparasitized carrot plants. Roots of parasitized plants tended to have similar or lower amino acid concentrations than roots of nonparasitized plants. Dry and germinated broomrape seeds had similar amino acid profiles, but individual amino acid concentrations were lower than in the other broomrape tissues examined. The broomrape shoot tended to have lower amino acid concentrations than the tubercle and callus. Free amino acid profiles of leaves and roots of parasitized plants paralleled those of nonparasitized plants, respectively. Individual free amino acids tended to occur at similar or lower levels in dry and germinated broomrape seeds than in the tubercle, shoot, or callus. Free amino acid composition of the broomrape tubercle was similar to that of the parasitized root. Arginine and alanine concentrations in broomrape callus were dramatically higher than those of other amino acids in this or other tissues investigated. These results indicate that changes in the composition of both free and bound amino acids in carrot are associated with broomrape parasitism.

Flatpea intoxication in sheep and indications of ruminal adaptation.

This paper describes the signs of toxicity when seed-bearing flatpea (Lathyrus sylvestris L) hay is fed to sheep. Signs of intoxication (including seizure, muscular trembling and spasmotic torticollis) are similar to those observed for ammonia toxicity in ruminants. Accumulation of ammonia may be a direct consequence of flatpea ingestion, given that 2,4-diaminobutyric acid (DABA, a toxic constituent of flatpea) is known to inhibit hepatic urea synthesis. However, other modes of toxicity for DABA as well as other flatpea toxins may also contribute to this process of intoxication. Our evidence suggests that ruminal microbes are responsible for flatpea detoxification and host animal protection. The adaptation of sheep to flatpea may be a consequence of enhanced ruminal detoxification activity. Ruminal protective functions can be disrupted, however, through abrupt monensin feeding or the replacement of nonadapted for adapted rumen contents. This disruption temporarily suppresses mechanisms of ruminal detoxification. As a consequence sheep can again be made vulnerable to flatpea intoxication.

The toxic effects of mature flatpea (Lathyrus sylvestris L cv Lathco) on sheep.

The toxic effects of mature, seed-bearing flatpea (Lathyrus sylvestris L cv Lathco) hay on sheep was studied in a feeding trial with 25 adult ewes. Five ewes were barren; 20 were in the last 10 w of pregnancy. The ewes were blocked by weight and reproductive status for assignment to treatment groups. Four groups of 5 ewes each were allotted to 1 of the following diets: 0% (controls), 35.0%, 52.5% or 70.0% flatpea (FP); and 1 group of 5 ewes was allotted to a diet of increasing levels of 17.5, 35.0, 52.5 and 70.0% FP. Flatpea hay was harvested in the mature, seed-bearing stage, pelleted, reground and incorporated into a basal (control) diet of 30% corn + 70% alfalfa by substitution of an appropriate amount of FP for alfalfa. The resulting diets were fed for up to 42 days. The toxic nature of FP was expressed by the following dose-related responses: Fluctuating reductions in feed intake upon initiation of diets containing 35% FP and above; occurrence of a potentially fatal, acute central nervous system syndrome at FP levels as low as 35%; and reduced weight gain at FP levels of 52.5% and above. A wasting condition was observed in 1 ewe fed 52.5% FP. Four of 5 ewes and 1 lamb fed increasing levels of FP developed acute poisoning. One of these ewes developed acute neurotoxicosis when consuming 52.5% FP; 3 other ewes and 1 40-d-old orphaned lamb developed neurotoxicosis upon raising the FP level to 70.0%.(ABSTRACT TRUNCATED AT 250 WORDS)

Apparent digestibility and nutrient balance in lambs fed different levels of flatpea hay.

Apparent digestibility and nutrient utilization were studied in a digestion and balance trial with 30 wether lambs (BW 32 kg). Lambs were blocked by weight and allotted randomly to five diets with ratios of alfalfa (Medicago sativa L.) to 'Lathco' flatpea (Lathyrus sylvestris L.) hay of 100:0, 75:25, 50:50, 25:75, and 0:100. Alfalfa was harvested in the early bloom stage and flatpea was harvested in the vegetative stage. Digestibilities of DM, NDF, ADF, cellulose, hemicellulose, and energy decreased linearly (P less than .05) as the level of flatpea hay increased to values of 53.3, 32.4, 39.9, 46.0, 40.5, and 52.5%, respectively, for 100% flatpea hay. Nitrogen retention (9 to 15% of intake) was not affected by level of flatpea hay. Ruminal pH, NH3 N, and blood urea N increased linearly (P less than .001) as level of flatpea increased in the diet, apparently a reflection of dietary N level. Calcium excretion decreased linearly (P less than .01) with increased proportions of flatpea hay and was related to dietary intake of Ca. Apparent absorption and retention showed a cubic effect (P less than .05). No clinical signs of toxicity were observed in any of the lambs during the 20-d metabolism trial. Vegetative flatpea is potentially valuable as a forage for feeding ruminants.

Cutaneous horn of penis.

Cutaneous horns of the penis are hyperkeratotic lesions of the skin which arise secondary to chronic irritation. Treatment is local excision with removal of a broad base. Examination of the base is important to rule out carcinoma as the underlying cause of the lesion. A case report is presented and the literature reviewed.

Intracellular Localization of the Neurotoxin 2,4-Diaminobutyric Acid in Lathyrus sylvestris L. Leaf Tissue.

The intracellular distribution of the neurotoxin 2,4-diaminobutyric acid (DABA) in mature leaves of the perennial legume Lathyrus sylvestris L. var ;Lathco' (flatpea) was determined using subcellular fractions from mesophyll protoplasts. Chloroplasts contained about 15% of the cellular DABA. At least 75% of the DABA was vacuolar, based on the assumptions that each protoplast contained a single vacuole and that acid phosphatase occurred exclusively in the vacuole. DABA was not detectable in peroxisomal and mitochondrial fractions. Because the vacuole is not a major site of amino acid synthesis, this distribution implicates synthesis of DABA within chloroplasts with subsequent transport to and storage within the vacuoles of the mesophyll cells.

Production and Use of Monoclonal Antibodies for Identification of Strains of Rhizobium trifolii.

We produced a monoclonal antibody against Rhizobium trifolii 162x95. This antibody in cell culture supernatant was used in an indirect enzyme-linked immunosorbent assay to differentiate strain 162x95 from naturalized strains in the Appalachian region. Nodules crushed in 0.1 to 0.2 ml of phosphate-buffered saline and used to charge enzyme-linked immunosorbent assay plates gave strong absorbance readings. Heat-inactivated and noninactivated portions of 162x95 cultures were strongly reactive, indicating that the antigen is probably a carbohydrate. Of 10 strains from California, where 162x95 was isolated, 6 strongly cross-reacted with the antibody. The cellular protein patterns in a sodium dodecyl sulfate-polyacrylamide gradient gel of cross-reactive strains were essentially identical. A Western blot analysis indicated that the antibody was against a 19.8-kilodalton band. The Western blot analysis also revealed that the polyvalent antiserum contained other strongly reacting antibodies with molecular weights of approximately 20,000, indicating the possibility that other monoclonal antibodies to detect strain 162x95 may be produced. However, the available antibody has been shown to be useful for short-term experiments. Based upon protein profiles and immunological reactions, there are 4 or 5 California strains rather than 10.

2,8-dihydroxyadenine urolithiasis: review of the literature and report of a case in the United States.

Urolithiasis resulting from inherited metabolic derangement is rare. Only 13 cases of 2,8-dihydroxyadenine stones resulting from a deficiency of the enzyme adenine phosphoribosyl transferase have been reported since 1974. Of these cases 9 have been in children with the homozygous trait. To date, 3 homozygous and 1 heterozygous adults with urolithiasis have been reported. This disease has not been associated with any other clinical or biochemical abnormalities. Treatment includes low purine diet and allopurinol. We herein report a case of complete adenine phosphoribosyl transferase deficiency associated with 2,8-dihydroxyadenine urolithiasis in the United States, bringing the total to 14 in the literature.

Nosocomial Pseudomonas aeruginosa urinary tract infections.

Two separate outbreaks of Pseudomonas aeruginosa urinary tract infections (UTIs) were associated with cystoscopy or transurethral prostate resection. The first outbreak was identified after routine bacteremia surveillance demonstrated four cases of P aeruginosa septicemia in a three-month period. A six-month retrospective review of the microbiology records identified 14 cases of P aeruginosa UTI associated with urologic surgery instrumentation. The outbreak terminated after the implementation of two major control measures: (1) replacement of hexachlorophene solution with an iodophor solution for preparing patients and cleaning instruments before disinfection, and (2) weekly gas sterilization of cystoscopy instruments. The second outbreak, consisting of 11 cases of P aeruginosa UTI after transurethral resection of the prostate gland, occurred in a 187-bed community hospital. All available patient isolates were serotype 011, and culture of a rubber adaptor attached to the resectoscope also yielded growth of that serotype. The outbreak promptly terminated when the rubber adaptor was sterilized between cases.

Intracellular Localization of Enzymes of Carbon Metabolism in Mesembryanthemum crystallinum Exhibiting C(3) Photosynthetic Characteristics or Performing Crassulacean Acid Metabolism.

Mesembryanthemum crystallinum, a halophilic, inducible Crassulacean acid metabolism (CAM) species, was grown at NaCl concentrations of 20 and 400 millimolar in the rooting medium. Plants from the low salinity treatment showed exclusively C(3)-photosynthetic net CO(2) fixation, whereas plants exposed to the high salinity level exhibited net CO(2) dark fixation involving CAM. Mesophyll protoplasts, isolated from both tissues, were gently ruptured, and the intracellular localization of enzymes was studied following differential centrifugation and Percoll density gradient centrifugation of protoplast extracts. Both centrifugation techniques resulted in the separation of intact chloroplasts, with up to 90% yield, from other organelles and the nonparticulate fraction of cells. Enzymes were identified by determination of activity and by sodium dodecyl sulfate gel electrophoresis of enzyme protein.Experiments established the extraorganellar (cytoplasmic) location of phosphoenolpyruvate carboxylase, enolase, phosphoglyceromutase, and NADP-malic enzyme; the mitochondrial location of NAD-malic enzyme; and the chloroplastic location of pyruvate, Pi dikinase. NAD-glyceraldehyde-3-phosphate dehydrogenase, phosphohexose isomerase, and phosphoglycerate kinase were associated with both cytoplasm and chloroplasts. NADP-dependent malate dehydrogenase activity was found in both the chloroplastic and extrachloroplastic fractions; the activity in the chloroplast showed an optimum at pH 8.0 and was dependent upon preincubation of enzyme with dithiothreitol. The extrachloroplastic activity showed an optimum at pH 6.5 and was independent of pretreatment with dithiothreitol. Protoplast extracts of M. crystallinum performing CAM exhibited higher activities (expressed per mg chlorophyll per min) of phosphoenolpyruvate carboxylase, pyruvate, Pi dikinase, NADP-malic enzyme, NAD-malic enzyme, NADP-malate dehydrogenase, enolase, phosphoglyceromutase, NAD-glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and phosphohexose isomerase than protoplast extracts from M. crystallinum not exhibiting CAM. The increase in total activity of the latter three enzymes following exposure of plants to 400 millimolar NaCl and the development of CAM was due to specific increases in the levels of activity in the cytoplasm.

Activity and quantity of ribulose bisphosphate carboxylase-and phosphoenolpyruvate carboxylase-protein in two Crassulacean acid metabolism plants in relation to leaf age, nitrogen nutrition, and point in time during a day/night cycle.

Activity of ribulose 1,5-bisphosphate (RuBP) carboxylase in leaf extracts of the constitutive Crassulacean acid metabolism (CAM) plant Kalanchoe pinnata (Lam.) Pers. decreased with increasing leaf age, whereas the activity of phosphoenolpyruvate (PEP) carboxylase increased. Changes in enzyme activities were associated with changes in the amount of enzyme proteins as determined by immunochemical analysis, sucrose density gradient centrifugation, and SDS gel electrophoresis of leaf extracts. Young developing leaves of plants which received high amounts of NO 3 (-) during growth contained about 30% of the total soluble protein in the form of RuBP carboxylase; this value declined to about 17% in mature leaves. The level of PEP carboxylase in young leaves of plants at high NO 3 (-) was an estimated 1% of the total soluble protein and increased to approximately 10% in mature leaves, which showed maximum capacity for dark CO2 fixation. The growth of plants at low levels of NO 3 (-) decreased the content of soluble protein per unit leaf area as well as the extractable activity and the percentage contribution of both RUBP carboxylase and PEP carboxylase to total soluble leaf protein. There was no definite change in the ratio of RuBP carboxylase to PEP carboxylase activity with a varying supply of NO 3 (-) during growth. It has been suggested (e.g., Planta 144, 143-151, 1978) that a rhythmic pattern of synthesis and degradation of PEP carboxylase protein is involved in the regulation of ß-carboxylation during a day/night cycle in CAM. No such changes in the quantity of PEP carboxylase protein were observed in the leaves of Kalanchoe pinnata (Lam.) Pers. or in the leaves of the inducible CAM plant Mesembryanthemum crystallinum L.

Responses of superoxide dismutase and glutathione reductase activities in cotton leaf tissue exposed to an atmosphere enriched in oxygen.

Responses of superoxide dismutase (EC 1.15.1.1) and glutathione reductase (EC 1.6.4.2) activities were evaluated in leaf tissue from intact cotton plants (Cotton Branch 1697) which were exposed to 75% O(2), 350 microliters per liter CO(2) for 48 hours. Soluble protein was extracted from O(2)-treated and control tissue, and enzyme levels were determined. Superoxide dismutase activity in cotton leaf tissue was high (26 units per milligram protein) under normal conditions of 21% O(2), saturating light, and limiting CO(2), and neither qualitative nor quantitative differences in the cyanide-sensitive or -insensitive forms of the enzyme occurred in response to hyperoxic conditions. Glutathione reductase activity, however, was 2- to 3-fold higher in extracts from tissue exposed to 75% O(2). No increase in activity was observed for the peroxisomal enzymes, glycolate oxidase (EC 1.1.3.1) and catalase (EC 1.11.1.6). Results are consistent with an integrated pathway involving superoxide dismutase and glutathione reductase for protection of sensitive leaf components against detrimental effects of intermediate reduction products of O(2).