Clinical utility of miRNA-1, miRNA-29g and miRNA-133s plasma levels in prostate cancer patients with high-intensity training after androgen-deprivation therapy.

The randomized trials showed that the addition of training resistance program to androgen-deprivation therapy (ADT) had many beneficial effects for prostate cancer (PC) patients (significant protective effect on the volume of muscle mass) and the studies have revealed a panel of miRNAs, which are deregulate in PC and may serve as promising biomarkers of PC risk. The primary aim of our present study was to investigate the effect of exercise training to changes in body composition (muscle strength) and the secondary endpoint was to investigate the impact of an exercise training program on plasma levels of selected myogenic microRNAs (miRNAs) (miRNA-1, miRNA-29b, and miRNA-133) in PC patients undergoing the ADT. Effect of ADT and exercise intervention showed significant increase (experimental group vs. control group) the changes in body composition, free testosterone levels, IL-6 and plasma levels of myogenic miRNAs and significant reduced insulin serum levels. In conclusion, resistance training with ADT in the treatment of PC significantly changed the physical and metabolic function and the plasma levels of specific myogenic miRNAs. Our data support with the other publicized results.

Reliability and methodological issues of power assessment during chest presses on unstable surface with different weights.

AIM: The study compares the reliability of peak power (Ppeak) and mean power in acceleration (Pmean acc) and entire concentric phase (Pmean total) of chest presses on the bench and unstable Swiss ball with different weights. METHODS: A group of 32 fit men performed over 2 testing sessions 3 trials of barbell chest presses on the bench and Swiss ball, without and with countermovement, with weights of 40, 60 and 80% 1RM. RESULTS: High values of correlation coefficients (above .80) and no significant differences between trials signify stability of measurement under both stable and unstable conditions. When chest presses were performed on the bench, ICC and SEM% values were in range .97 to .98 and 7.6 to 7.7%, respectively for Pmean total, .96 to .98 and 9.1 to 9.6%, respectively for Pmean acc, and .94 to .97 and 9.2 to 10.0%, respectively for Ppeak. Their values during chest presses on a Swiss ball ranged from .93 to .96 and 8.4 to 9.1%, respectively for Pmean total, from .87 to .90 and 11.7 to 12.2%, respectively for Pmean acc, and from .79 to .82 and 12.1 to 13.4%, respectively for Ppeak at weights of 40 and 60% 1RM, and from .70 to .76 and 17.6 to 19.8%, respectively at weight of 80% 1RM. CONCLUSION: Measurement of peak and mean power during unstable chest presses provides reliable data comparable to those obtained during bench presses under all conditions tested. However, peak values of power measured during unstable chest presses with weights ≥80% 1RM should be interpreted with caution.

Utilization of elastic energy during weight exercises differs under stable and unstable conditions.

AIM: The study compares the power outputs in concentric phase of chest presses and squats performed without and with countermovement on stable and unstable support surface, respectively. METHODS: A group of 16 physical education students performed randomly in four different days 3 repetitions of: 1) barbell chest presses on the bench and Swiss ball, respectively, and 2) barbell squats on stable support base and Bosu ball, respectively. Exercises were performed without and with countermovement (CM) using maximal effort in concentric phase of lifting. Initial weight of 20 kg increased by 10 kg or 5 kg (at higher loads) up to at least 85% of previously established 1RM under stable conditions. A PC based system FiTRO Dyne Premium was used to monitor force and velocity and to calculate power. As a parameter of the capability to use elastic energy was considered the difference in mean power in concentric phase of weight exercises (ΔP) performed with and without CM. RESULTS: There were higher power outputs in concentric phase of CM chest presses as compared to those performed from quiet position on the chest under both stable and unstable conditions. This enhancement of power due to CM was rather modest at lower weights and become more pronounced with increasing weights reaching a maximum at 57.1% 1RM on stable and at 47.6% 1RM on unstable support surface. Lifting heavier weights not only failed to increase the enhancing effect but led to its decline. Similar trend was observed during squats with maximal enhancement of power in concentric phase of lifting at about 80% 1RM under both conditions. The ΔP was significantly lower during chest presses on Swiss ball than on the bench, however, only at higher weights lifted (≥60% 1RM). On the other hand, the ΔP during squats performed on Bosu ball and on stable support base did not differ significantly across all weights lifted. CONCLUSION: The ability to utilize elastic energy during CM chest presses is more profoundly compromised under unstable than stable conditions, namely at higher weights lifted. On the other hand, there is similar enhancement of power in concentric phase of CM squats on stable and unstable support surface regardless of weights lifted. Besides the type of exercise, this may be ascribed to different degree of instability of devices used (Swiss ball vs. Bosu ball).

The effect of 6-week combined agility-balance training on neuromuscular performance in basketball players.

AIM: The study evaluates the effect of 6-week combined agility-balance training on neuromuscular performance in basketball players. METHODS: Subjects divided into experimental (EG, n = 17) and control group (CG, n = 17) underwent a combined agility-balance training (in duration of 30 min) for a period of 6 weeks (4-5 sessions/week). Both groups performed reaction tasks similar to game-like situations, however EG on wobble boards and CG on stable surface. Prior to and after the training parameters of agility, balance, speed of step initiation, strength differentiation accuracy, and explosive power of lower limbs were evaluated. Postural stability was assessed under both static and dynamic conditions (wobble board) with eyes open and eyes closed, respectively. The velocity of the centre of pressure (COP) was registered at 100 Hz by means of posturography system FiTRO Sway check based on dynamometric platform. Using FiTRO Reaction check simple and multi-choice reaction times were measured. The same system was applied to evaluate the agility performance including reaction and movement task. Speed of step initiation was measured using FiTRO Dyne Premium. Jumping abilities were evaluated by means of FiTRO Jumper (10-seconds maximal jumps, Countermovement jump, Squat jump, Drop jump). Using the same system, the subject´s ability to match 50 % of their maximal height of the jump was evaluated. RESULTS: Results showed that a combined agility-balance training improved dynamic balance not only under visual control but also in eyes closed conditions. Training also increased run-out speed that likely contributed to better agility performance, reduced ground contact time during drop jump, and improved the ability to differentiate the force of muscle contraction during repeated jumps. However, such training has been found to be insufficient to improve both simple and multi-choice reaction time, and jumping performance. On the other hand, control group failed to show any significant improvement in examined abilities except for enhancement of jumping performance (Pact, Δ CMJ & SJ). CONCLUSION: It may be concluded that balance exercises performed simultaneously with reaction tasks represent an effective means for improvement of neuromuscular performance in elite athletes.

Effect of head-down bed rest on the neuroendocrine response to orthostatic stress in physically fit men.

The role of neuroendocrine responsiveness in the development of orthostatic intolerance after bed rest was studied in physically fit subjects. Head-down bed-rest (HDBR, -6 degrees, 4 days) was performed in 15 men after 6 weeks of aerobic training. The standing test was performed before, after training and on day 4 of the HDBR. Orthostatic intolerance was observed in one subject before and after training. The blood pressure response after training was enhanced (mean BP increments 18+/-2 vs. 13+/- 2 mm Hg, p<0.05, means +/- S.E.M.), although noradrenaline response was diminished (1.38+/-0.18 vs. 2.76+/-0.25 mol.l(-1), p<0.01). Orthostatic intolerance after HDBR was observed in 10 subjects, the BP response was blunted, and noradrenaline as well as plasma renin activity (PRA) responses were augmented (NA 3.10+/-0.33 mol.l(-1), p<0.001; PRA 2.98+/-1.12 vs. 0.85+/-0.15 ng.ml(-1), p<0.05). Plasma noradrenaline, adrenaline and aldosterone responses in orthostatic intolerant subjects were similar to the tolerant group. We conclude that six weeks of training attenuated the sympathetic response to standing and had no effect on the orthostatic tolerance. In orthostatic intolerance the BP response induced by subsequent HDBR was absent despite an enhanced sympathetic response.

Effect of simulated microgravity on endocrine response to insulin-induced hypoglycemia in physically fit men.

Adaptation to microgravity is associated with alteration in some endocrine functions. In the present longitudinal study, the counterregulatory hormonal response to insulin-induced hypoglycemia (ITT, 0.1 IU/kg short acting insulin i. v.) was evaluated under simulated microgravity conditions in 15 physically fit subjects. ITT was performed at the beginning of the investigation, and again after completion of 6 weeks of endurance training and after a subsequent period of 4 days of head-down bed rest at a backward tilt of 6 degrees from the horizontal. Endurance training showed a significant increase in maximal aerobic capacity in previously well-trained subjects (increase by 12 %), as well as on attenuation of counterregulatory response of epinephrine to hypoglycemia. After 4 days of bed rest, basal concentrations of plasma norepinephrine was diminished (p < 0.002) and plasma renin activity was enhanced (p < 0.02). After bed rest, decreased responses of the two catecholamines (norepinephrine, p < 0.001; epinephrine, p < 0.001), growth hormone (p < 0.001), and cortisol (p < 0.05) were observed. Response of plasma renin activity after bed rest was increased (p < 0.01). This longitudinal study indicated that 4 days of bed rest in endurance-trained subjects induced increased response of PRA to hypoglycemia and attenuation of other counterregulatory neuroendocrine responses.

Effects of dietary cholesterol restriction in a feline model of Niemann-Pick type C disease.

A feline model of Niemann-Pick disease type C (NPC) was employed to evaluate the effect of dietary cholesterol restriction on progression of disease. Two NPC-affected treated cats were fed a cholesterol-restricted diet beginning at 8 weeks of age; the cats remained on the diet for 150 and 270 days respectively. The study goal was to lower the amount of low density lipoprotein (LDL) available to cells, hypothetically reducing subsequent lysosomal accumulation of unesterified cholesterol and other lipids. Neurological progression of disease was not altered and dietary cholesterol restriction did not significantly decrease storage in NPC-affected treated cats. One NPC-affected treated cat had decreased serum alkaline phosphatase activity (ALP) and decreased serum cholesterol concentration. Liver lipid concentrations of unesterified cholesterol, cholesterol ester and phospholipids in NPC-affected treated cats were similar to those seen in NPC-affected untreated cats. Ganglioside concentrations in the NPC-affected treated cats and NPC-affected untreated cats were similar. Histological findings in liver sections from NPC-affected treated cats showed a diffuse uniform microvacuolar pattern within hepatocytes and Kupffer cells, in contrast to a heterogeneous macro/microvacuolar pattern and prominent nodular fibrosis in NPC-affected untreated cats. Similar differences in vacuolar patterns were seen in splenic macrophages. Although some hepatic parameters were modified, dietary cholesterol restriction did not appear to alter disease progression in NPC-affected kittens.

Serum zinc, chromium, and iron concentrations in dogs with lymphoma and osteosarcoma.

We compared serum concentrations of zinc, chromium, and iron in dogs with cancer to those of normal dogs. Dogs with lymphoma (n = 50) and osteosarcoma (n = 52) were evaluated. Dogs with lymphoma had significantly lower (P = .0028) mean serum zinc concentrations (mean +/- SD; 1.0 +/- 0.3 mg/L) when compared to normal dogs (1.2 +/- 0.4 mg/L). Dogs with osteosarcoma also had lower mean serum zinc concentrations (1.1 +/- 0.4 mg/L), but this difference was not significant (P = .075). Serum chromium concentrations were significantly lower in dogs with lymphoma (2.6 +/- 2.6 microg/L, P = .0007) and osteosarcoma (2.4 +/- 3.1 microg/L, P = .0001) compared to normal dogs (4.7 +/- 2.8 microg/L). Serum iron concentrations and total iron-binding capacity were significantly lower in dogs with lymphoma (110.8 +/- 56.7 microg/dL, P < .0001, and 236.6 +/- 45.6 microg/dL, P < .0001, respectively) and osteosarcoma (99.6 +/- 49.3 microg/dL, P < .0001, and 245.0 +/- 43.8 microg/dL, P = .0011, respectively) when compared to normal dogs (175.1 +/- 56.7 microg/dL and 277.1 +/- 47.4 microg/dL). Mean ferritin concentration was significantly higher in dogs with lymphoma (1291.7 +/- 63.0 microg/L) than in normal dogs (805.8 +/- 291.1 microg/L, P < .0001) and dogs with osteosarcoma (826.5 +/- 309.2 microg/L, P < .0001). Further investigation is needed to explore the clinical significance of these mineral abnormalities in dogs with cancer.

Inhibition of canine and feline alcohol dehydrogenase activity by fomepizole.

OBJECTIVE: To determine and compare substrate specificity and kinetic rate constants of feline and canine alcohol dehydrogenase (ADH) with ethanol (EtOH) and ethylene glycol (EG) as substrates in vitro, with and without fomepizole. SAMPLE POPULATION: Livers from 3 dogs and 3 cats. PROCEDURE: Canine and feline ADH activity, in cytosolic fractions of homogenized liver, was determined by use of various concentrations of nicotinamide adenine dinucleotide (NAD), EtOH, or EG as substrates. Initial reaction velocities were calculated, and kinetic inhibition rate constants (Ki) for fomepizole were determined. RESULTS: Substrate specificity of canine and feline ADH for EtOH or EG was not significantly different. A 2-fold difference was detected in the maximal velocity of canine, compared with feline, ADH, using either substrate. Fomepizole Ki in feline hepatic homogenates was significantly greater than Ki in canine hepatic homogenates when either EtOH or EG was used as substrate (10- and 30-fold, respectively). A 6-fold increase in the concentration of fomepizole was required to achieve ADH inhibition, with feline homogenates equivalent to those of canine homogenates. CONCLUSIONS AND CLINICAL RELEVANCE: Feline ADH has lower enzymatic capacity for turnover or is less concentrated in liver than canine ADH with regard to EtOH and EG catalysis. Canine ADH was more effectively inhibited by fomepizole than feline ADH. Results suggest that higher dosages of fomepizole may be more effective to treat cats with EG intoxication than dosages reported to treat dogs.

Effects of dietary cysteine on blood sulfur amino acid, glutathione, and malondialdehyde concentrations in cats.

OBJECTIVE: To determine effects of dietary cysteine on blood sulfur amino acids (SAA), reduced glutathione (GSH), oxidized glutathione (GSSG), and malondialdehyde (MDA) concentrations in cats. ANIMALS: 12 healthy adult cats. PROCEDURE: Cats were fed diets with a nominal (0.50 g/100 g dry matter [DM]), moderate (1.00 g/100 g DM), or high (1.50 g/100 g DM) cysteine content in a 3 X 3 Latin square design with blocks of 8 weeks' duration. Venous blood samples were collected after each diet had been fed for 4 and 8 weeks, and a CBC and serum biochemical analyses were performed; poikilocyte, reticulocyte, and Heinz body counts were determined; and MDA, GSH, GSSG, and SAA concentrations were measured. RESULTS: Blood cysteine and MDA concentrations were not significantly affected by dietary cysteine content. Blood methionine, homocysteine, and GSSG concentrations were significantly increased when cats consumed the high cysteine content diet but not when they consumed the moderate cysteine content diet, compared with concentrations obtained when cats consumed the nominal cysteine content diet. Blood GSH concentrations were significantly increased when cats consumed the moderate or high cysteine content diet. CONCLUSIONS: Increased dietary cysteine content promotes higher blood methionine, homocysteine, GSH, and GSSG concentrations in healthy cats. CLINICAL RELEVANCE: Supplemental dietary cysteine may be indicated to promote glutathione synthesis and ameliorate adverse effects of oxidative damage induced by disease or drugs.

Association of excess sulfur intake and an increase in hydrogen sulfide concentrations in the ruminal gas cap of recently weaned beef calves with polioencephalomalacia.

During a 2-week period, 16 of 150 recently weaned calves developed signs of polioencephalomalacia (PEM). One calf was examined and treated at our veterinary teaching hospital and a necropsy was performed on a calf that died. During the peak of the outbreak, clinicians visited the ranch. Ruminal hydrogen sulfide (H2S) and blood thiamine concentrations were measured in 10 clinically normal penmates of PEM-affected calves. Ruminal H2S concentrations were high (> 0.695 mg/L [> 500 ppm]) in all cattle (mean, 12.19 mg/L [8,770 ppm]). All blood thiamine values were within the reference range. Within 12 hours after measurement of blood thiamine concentrations, 2 of the calves from which samples were obtained developed clinical signs of PEM. Dietary analysis revealed an estimated sulfur intake of 0.9% per calf on a dry-matter basis. Hay contributed most of this sulfur. In the investigation reported here, an outbreak of PEM was associated with high ruminal H2S concentrations and excess sulfur intake without evidence of thiamine deficiency.

Effects of weight gain and loss on metabolic rate, glucose tolerance, and serum lipids in domestic cats.

Weight gain is a common problem in domestic cats, but little is known about its metabolic effects. The purpose of this study was to determine the effects of diet-induced weight gain and subsequent weight loss on metabolic rate, body composition, and glucose tolerance. Gain of approximately 20 per cent body weight (divided approximately equally between fat and fat-free mass) over three months resulted in insulin resistance in females, indicated by increases in basal insulin concentration (68.2+/-7.9 to 119+/-16.5 pmol litre(-1), P<0.05), insulin peak response to glucose (241.1+/-31.6 to 315.0+/-23.0 pmol litre(-1), P<0.05), and deltaI/deltaG (14.2+/-2.6 to 18.1+/-1.3 pmol mmol(-1), P<0.05) compared with pre-gain values. The same numerical trend was noted in male cats, however, changes were not significant (P>0.05). Alterations in serum lipids included significant (P<0.05) elevations in triglyceride concentrations in male cats and decreased beta-lipoprotein concentrations in both genders. Weight loss over three months normalised basal insulin, insulin response to glucose, and serum triglyceride concentrations, and resulted in significant (P<0.05) decreases in serum concentrations of beta- and prebeta-lipoproteins, cholesterol, and triiodothyronine. Diet-induced weight gain of three months' duration, followed by three months' maintenance of increased body weight did not affect fasting or resting metabolic rate. Development and severity of impaired glucose tolerance, insulin resistance, and other changes may be affected by duration and possibly severity of weight gain.

Mechanisms of echinocytosis induced by Crotalus atrox venom.

Transient echinocytosis has been reported in association with snake envenomation in humans and dogs. An in vitro model of echinocytosis induced by venom of crotalus atrox (western diamondback rattlesnake) was established to characterize erythrocyte morphologic changes and to investigate potential mechanisms of echinocytic transformation. Erythrocyte morphologic changes produced after the addition of venom to canine, feline, equine, and human blood were characterized by dose-dependent echinocytosis. Type III echinocytosis were consistently induced in vitro at a dose comparable to in vivo envenomation; higher venom doses produced spheroechinocytic and spherocytic transformations. The changes could not be induced in vitro in the presence of ethylenediaminetraacetic acid but were observed in heparinized and citrated blood samples, suggesting the participation of calcium or a metalloprotein in echinocytic change. These findings suggest that phospholipase A2 (PLA2), a calcium-dependent enzyme in snake venom, may be responsible for echinocytic transformation via the production of lysolecithin, a known echinocytic agent. Purified PLA2 from C. atrox venom induced dose-dependent echinocytic change in vitro in canine blood. Other potential mechanisms of echinocytic change evaluated in canine blood included erythrocyte cation loss and erythrocyte ATP depletion. In canine blood mixed with venom, erythrocyte sodium and potassium concentrations were consistently less than those of controls, likely as a result of membrane alteration produced by the actions of PLA2. There was no difference in blood ATP concentrations from dogs with snakebite when compared with normal controls; however, the power of this comparison was low. Echinocytosis induced by rattlesnake venom is related to the degree of venom exposure and may correlate clinically with the amount of venom absorbed. Echinocytic transformation in vitro is induced by PLA2 present in venom.

Effects of neutering on bodyweight, metabolic rate and glucose tolerance of domestic cats.

Few controlled studies have been made of the possible mechanisms and physiological consequences of weight gain after cats have been neutered. In this study, six male and six female cats were gonadectomised and compared with five entire male and six entire female cats, before they were neutered and one and three months later. The neutered males gained significantly more weight (mean [SEM] per cent) than the entire males (30.2 [5.2] v 11.8 [2.3]) and the entire females gained 40.0 (7.3) v 16.1 (3.3) per cent, (P < 0.05). The castrated males gained more weight as fat than the sexually intact males (22.0 [3.3] v 8.8 [4.5] per cent, P < 0.05). There was a significant increase (P < 0.05) in daily food intake after neutering. Spayed females underwent a significant decrease in fasting metabolic rate (83.7 [5.5] v 67.2 [2.3] kcal/kg bodyweight0.75/day P < 0.05). Gonadectomy had minimal effects on serum thyroid hormone concentrations, the resting or fasting metabolic rates in males, or on indices of glucose tolerance.

In vivo indicators of pathologic ruminal sulfide production in steers with diet-induced polioencephalomalacia.

Two groups of 3 120-160-kg Holstein steers were fed a diet high in carbohydrate and low in long fiber and either with or without added sodium sulfate. Prior to and during the course of feeding the experimental diet, the concentrations of rumen hydrogen sulfide gas and rumen fluid sulfide were determined by a simple sulfide detector tube method and by sulfide-selective electrode, respectively. Other measurements included rumen fluid pH, blood creatine kinase, and blood sulfhemoglobin. Two of the 3 steers fed the high-sulfate diet developed signs and lesions of polioencephalomalacia. Clinical signs included episodic ataxia and blunted or absent menace reaction. Increased ruminal H2S gas concentrations occurred in all 3 steers consuming the diet with added sulfate. The onset of clinical signs coincided with the onset of elevated H2S concentrations. These increases were 40-60 times the values measured in the steers consuming the diet without added sulfate. In contrast, increases in rumen fluid sulfide concentrations usually rose to 4 times that of control steers. The steers fed an identical diet but without added sulfate exhibited no signs or lesions of polioencephalomalacia and no elevations of sulfide in rumen gas or fluid. All steers had a modest decrease in rumen fluid pH associated with the transition to the concentrate diet. No significant changes were observed in any of the blood measurements of any of the steers. An additional pair of steers was fed the experimental diet with or without added sulfate to compare the ruminal H2S gas concentrations estimated by H2S detector tubes with those estimated by a different method of analysis utilizing charcoal trapping of H2S, conversion to sulfate, and measurement of the sulfate. Both methods yielded comparable estimates of H2S concentration. Overall, these data indicate that changes in rumen gas cap H2S concentrations are larger than changes in rumen fluid sulfide concentration and the estimation of rumen gas cap H2S concentration may be a practical approach to detecting pathologic increases in ruminal H2S gas. This simple, rapid, minimally invasive method should be useful for estimating the H2S content of ruminal gas under field conditions.

Safety and efficacy of 4-methylpyrazole for treatment of suspected or confirmed ethylene glycol intoxication in dogs: 107 cases (1983-1995).

OBJECTIVE: To evaluate safety and efficacy of 4-methylpyrazole (4-MP) treatment in dogs and to determine clinical signs and outcome of, and clinicopathologic abnormalities in, dogs treated in early or late stages of ethylene glycol (EG) intoxication. DESIGN: Retrospective study. ANIMALS: 107 dogs. PROCEDURE: For dogs treated with 4-MP, 1 of 2 dosage regimens was usually used: 20 mg/kg of body weight, IV, initially, 15 mg/kg 17 hours later, and 5 mg/kg 25 and 36 hours after the initial dose, or 20 mg/kg, IV, initially, 15 mg/kg 12 and 24 hours later, and 5 mg/kg 36 hours after the initial dose. RESULTS: Neither adverse clinical signs nor clinicopathologic abnormalities were associated with the administration of 4-MP except in 1 dog, which developed tachypnea, gagging, excess salivation, and trembling after the second dose of 4-MP was given. Ethylene glycol intoxication was confirmed in 37 dogs. Of these, 21 were azotemic or became azotemic within 18 hours after admission, and only 1 of the 21 survived. All 16 dogs that did not become azotemic survived. Median time from EG ingestion to treatment with 4-MP was 5 hours (range, 2 to 8.5 hours) for dogs that were not azotemic at admission and 14.5 hours (range, 8.5 to 38 hours) for dogs that were azotemic at admission. CLINICAL IMPLICATIONS: 4-MP was a safe and effective treatment for EG intoxication when it was given before sufficient quantities of EG had been metabolized to induce renal failure. Dogs treated within 8 hours of EG ingestion had a good prognosis.

Comparison of four methods for forage nitrate analysis.

Twenty forage samples were collected and selected for variation in nitrate content. Each forage samples was analyzed 4 times by 4 different methods: diphenylamine spot plate, spectrophotometric, nitrate-selective electrode, and high-performance liquid chromatographic. Five feed extracts were spiked with 2 different amounts of nitrate and analyzed by each method. The spectrophotometric and nitrate-selective electrode had similar percent recoveries, which were close to 100%. The nitrate-selective electrode method had the least variation of the 4 methods. The diphenylamine spot plate method had the poorest average recovery, greatest variation, and was the least accurate. The average coefficients of variation for all samples within a method were 15%, 12%, 6.4%, and 16 for the diphenylamine spot plate, spectrophotometric, nitrate-selective electrode, and high-performance liquid chromatographic methods, respectively. The variation in the nitrate-selective electrode method was lower (P < 0.05) than the other methods. The results from this study suggest that the nitrate-selective electrode method is more accurate and precise than the other methods of analysis tested.

Identity and interactions of rumen microbes associated with dietary sulfate-induced polioencephalomalacia in cattle.

To study their role in sulfate reduction, anaerobic bacteria were cultured from rumen fluid samples of cattle fed high-carbohydrate, short-fiber diets with and without added sulfate. The steers fed the diet with added sulfate developed polioencephalomalacia. Microbiological methods included colony type profiles, molybdate sensitivity, presence of desulfoviridin, sulfate reduction rates of pure and mixed cultures, and incubation time effects on sulfate reduction. Colony-type profiles indicated decreased diversity, but no relative change in numbers of sulfate-reducing bacteria in rumen fluid from cattle fed diets with and without added sulfate. Thirteen bacteria] isolates were selected for further study on the basis of colony type, sulfate-reducing activity, and growth in lactate, sulfate, and yeast extract media. Seven of the isolates had Desulfovibrio-like characteristics (ie, they were gram-negative, motile rods that reduced sulfate, were inhibited by molybdate, and contained the pigment desulfoviridin). The remaining 6 isolates were gram-negative, nonmotile rods. Four of these released sulfide from cysteine, and 2 generated only limited amounts of sulfide from sulfate or cysteine. The 7 sulfate reducing isolates generated sulfide in rumen fluid broth medium at greater rates than those observed in fresh rumen fluid. Sulfate reduction Could be sustained in cultures for prolonged incubation times if the gas phase containing hydrogen sulfide was replaced at frequent intervals. Variations in the amount of sulfate reduced by the pure cultures were most pronounced at short incubation times. Sulfate reduction was not inhibited in mixed cultures of sulfate-reducing and nonsulfate-reducing bacteria.

Ruminal microbial alterations associated with sulfide generation in steers with dietary sulfate-induced polioencephalomalacia.

Holstein steers were fed carbohydrate-rich, short-fiber basal diets with and without added sodium sulfate. Steers fed the high-sulfate diet developed the CNS disorder polioencephalomalacia (PEM). The onset of signs of PEM was associated with increased sulfide concentration in the rumen fluid. Over the course of the disease, anaerobic rumen bacteria were enumerated in roll tubes by use of the Hungate method Lo determine the effect of dietary sulfate on sulfate-reducing bacterial numbers. Media used included a general type for total counts and sulfate containing media with and without cysteine to assess sulfate-reducing bacteria. Changes in total and sulfate reducing bacterial numbers attributable to dietary sulfate content were not observed. The capacity to generate hydrogen sulfide from sulfate in fresh rumen fluid in vitro was substantially increased only after steers had been fed the high sulfate diet for 10 to 12 days, which coincided with the onset of signs of PEM. The low capacity for hydrogen sulfide production of rumen fluid taken at earlier times in the feeding period suggests that rumen microorganisms must adapt to higher dietary sulfate content before they are capable of generating potentially toxic concentrations of sulfide.