Effect of climatic factors on urinary cortisol and peripheral blood leukocytes in lambs grazing on a semi-natural grassland in the Hokuriku District of Japan.

The effects of climatic factors on stress and immune functions of grazing lambs in summer and autumn in the Hokuriku District of Japan were evaluated by determining urinary cortisol (U-COR) levels and peripheral blood leukocyte populations and comparing those with lambs kept indoors. Two groups of five lambs, consisting of those grazed on a semi-natural grassland (GRL) and those housed indoors in a domestic animal shelter (INL), were maintained at from July to October. The temperature-humidity index at each location was indicative of heat stress during summer; however, the U-COR elevation was not observed in both groups. The elevation was observed in GRL in autumn and was higher than INL in October. Climatic conditions in autumn were characterized by high humidity and a sudden drop in temperature. U-COR was positively correlated with the relative humidity. The GRL was exposed to low-nutrient conditions for a relatively long time. The CD4+ /CD8+ T cell ratio in GRL decreased in October. Subsequently, the total leucocyte, including granulocytes, monocytes, and lymphocytes, sharply increased. The responses indicated an immune deficiency caused by immunosuppression because of a low nutrition caused by grazing and high-stress conditions in autumn.

The Blood-Cerebrospinal Fluid Barrier Is Selective for Red Cabbage Anthocyanins and Their Metabolites.

The study aim was to determine whether strongly bioactive hydrophilic red cabbage anthocyanins possess the ability to cross the blood-cerebrospinal fluid barrier (blood-CSF barrier) and whether there is a selectivity of this barrier toward these compounds. To fulfill objectives, red cabbage preparation, containing nonacylated and acylated anthocyanins, was administered to 16 sheep with implanted cannulas into the brain third ventricle, and next, within 10 h, blood, urine, and the cerebrospinal fluid (CSF) were collected and analyzed with HPLC-MS/MS. Though, in blood plasma and urine after red cabbage intake, both, acylated and nonacylated anthocyanins and their metabolites occurred, but only nonacylated derivatives were present in the CSF, and their changes in the profile and concentration in the CSF resulted from the fluctuation of these pigments' concentration and profile in blood, their different abilities to permeate via the blood-CSF barrier, and their transformations in this barrier. Results indicate that the blood-CSF barrier is selective for red cabbage anthocyanins.

Comparison between the urine dipstick and the pH-meter to assess urine pH in sheep and dogs.

BACKGROUND: Urine pH is an integral part of a complete urinalysis, and is commonly measured in veterinary practice using semiquantitative reagent strips. OBJECTIVE: The aim of this study was to compare the urine pH of dogs and sheep, using visual interpretation of dipstick reactions, and using a pH-meter as the reference method. Agreement between the 2 methods was also assessed. An additional objective was to compare the urine pH before and after centrifugation. METHODS: A total of 50 voided urine samples from sheep and 52 from dogs were collected into sterile containers. For pH measurements, 2 methods were used, a pH-meter and urine dipstick reagent pads. Measurements were performed using urine samples before (whole urine) and after centrifugation (urine supernatant). For comparison of the 2 methods, Passing and Bablok regression analysis and Bland-Altman plots were used. RESULTS: The equation created to assess agreement between the 2 methods in dogs showed a constant bias at -0.14 and a positive proportional bias at 0.98. From a clinical standpoint, total bias was below and above the maximum acceptable bias in sheep and dogs, respectively. Clinically acceptable bias was also found using centrifuged urine samples in sheep, but the urine pH values before and after centrifugation were nearly identical in dogs. CONCLUSION: Urine dipstick reagent pads and pH-meters can be used interchangeably to determine urine pH in sheep without needing centrifugation. In contrast, pH-meters provide more accurate pH measurements than urine dipstick pads in canine urine, which is not improved by centrifugation.

Technical note: Evaluation of urinary purine derivatives in comparison with duodenal purines for estimating rumen microbial protein supply in sheep.

A data set of individual observations was compiled from digestibility trials to examine the relationship between the duodenal purine bases (PB) flow and urinary purine derivatives (PD) excretion and the validity of different equations for estimating rumen microbial N (Nm) supply based on urinary PD in comparison with estimates based on duodenal PB. Trials (8 trials, = 185) were conducted with male sheep fitted with a duodenal T-type cannula, housed in metabolic cages, and fed forage alone or with supplements. The amount of PD excreted in urine was linearly related to the amount of PB flowing to the duodenum ( < 0.05). The intercept of the linear regression was 0.180 mmol/(d·kg), representing the endogenous excretion of PD, and the slope was lower than 1 ( < 0.05), indicating that only 0.43% of the PB in the duodenum was excreted as PD in urine. The Nm supply estimated by either approach was linearly related ( < 0.05) to the digestible OM intake. However, the Nm supply estimated through either of 3 published PD-based equations probably underestimated the Nm supply in sheep.

Detection of residues in urine and tissues of sheep treated with trace levels of dietary ractopamine HCl.

Qualitative assays are sometimes used as the sole basis for detecting drug residues in live animals or in animal products. Such assays have become increasingly sensitive as detection technologies have improved, yet the limitations of such assays to discriminate purposeful and accidental drug exposures remain poorly defined. A study was conducted to determine the ability of a ractopamine lateral flow assay to accurately detect incurred ractopamine residues in contaminated feeds and in sheep fed trace quantities of ractopamine HCl. False positive and negative samples were determined using a quantitative liquid chromatography-tandem mass spectrometric (LC-MS/MS) method. Ractopamine HCl was fed to sheep at 0 (Zero), 1 (Low), 10 (Med), or 100 (High) µg/kg of diet ( = 4 per level, 0.5 kg of feed/d) for 7 consecutive d and urine was collected daily about ∼16 h post exposure. On-site lateral flow assays were able to reliably (0% false negatives) detect 20 µg of ractopamine HCl per kg of feed. Urine from treated sheep tested positive for ractopamine residues by lateral flow assay in 7.4 (Zero), 0 (Low), 82 (Med), and 86% (High) of the urine samples from each group. Parent ractopamine was below the assay limit of quantification (LOQ, 0.7 ng/mL) in all urine samples using LC-MS/MS. After hydrolysis of ractopamine conjugates, total ractopamine (parent + hydrolyzed metabolites) in urine of Low animals was always less than the LOQ, but in 7 of 28 samples were above the limit of detection (LOD, 0.22 ng/mL). In contrast, urine in Med animals contained 1.08 to 9.13 ng/mL of total ractopamine, while urine of High animals contained 4.85-32.82 ng/mL of total ractopamine. Ractopamine is rapidly eliminated; nevertheless, > 80% of urine samples from sheep exposed to 5 µg/d (M) of ractopamine HCl had detectable residues by the screening assay and a 100% of samples had measurable ractopamine using LC-MS/MS methods. Tissue residues of ractopamine were not detected in any of the sheep. The sensitivity with which the rapid, qualitative assay detected ractopamine was sufficient to reveal trace ractopamine exposures; these data suggest that the use of qualitative tests to indicate purposeful treatment of animals (i.e., for doping or growth enhancement), in the absence of collaborating quantitative data, is inappropriate.

Changes in the metabolic profile of pregnant ewes to an acute feed restriction in late gestation.

AIM: To detect early changes in the metabolic profile of pregnant ewes subject to acute feed restriction at 130 days of gestation, and to establish indicators of risk for ovine pregnancy toxaemia (OPT) for diagnostic purposes. METHODS: Twenty Corriedale ewes with known mating dates, carrying a single fetus, were used. Ewes were maintained on meadow grasslands and at 130 days of gestation were randomly divided in two groups of 10 ewes. The control group had ad libitum access to pasture. Ewes in the restricted group were subjected to an acute feed restriction for a maximum of 144 hours (6 days), with free access to water. From the start (0 hours) until the end of feed restriction, blood samples were collected from all ewes to monitor concentrations of cortisol, non-esterified fatty acids (NEFA), ß-hydroxybutyrate (BOHB) daily, and glucose in plasma every 6 hours; urinary pH was also measured. Every 6 hours the food restricted ewes were observed to detect clinical signs of OPT e.g. apathy, grinding teeth, empty chewing movements, head leaning against the wall, tachypnea and not drinking water. RESULTS: In food-restricted ewes, concentrations of glucose decreased and differed from control ewes from 54 to 90 hours (p<0.001), and 96 to 102 hours (p<0.05). Concentrations of BOHB, cortisol and NEFA increased following feed restriction and differed from control ewes after 48 to 144 hours (p<0.01). Eight of the 10 restricted ewes showed clinical signs of OPT after 102-132 hours. Mean concentrations of glucose, BOHB and cortisol differed between control and restricted ewes prior to the onset of clinical signs of OPT, after 48-96 hours of feed restriction (p<0.01). Mean gestational length, and time from birth to placental expulsion was not affected by the feed restriction. CONCLUSIONS: Our results suggest that concentrations of glucose, BOHB and cortisol in plasma may provide a precocious diagnosis of subclinical OPT, using values of 1.59 (SD 0.24) mmol/L, 2.26 (SD 1.03) mmol/L and 15.09 (SD 7.75) nmol/L, respectively. The identification of a potentially harmful metabolic imbalance could lead to the improvement of treatment success.

Effects of straw treatment and nitrogen supplementation on digestibility, intake and physiological responses of water intake as well as urine and faecal characteristics.

This study investigates the effects of feeding diet based on untreated (UT) or ensiled alkali-treated (ET) barley straw with either urea or casein supplementation, on feed intake, digestibility, ruminal pH, water intake and faecal and urinary characteristics. Four sheep fitted with ruminal cannulas were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Barley straw was treated by the dry (spraying) method in pH adjusted of hydrogen peroxide (pH 11.5), ensiled for 6 weeks and included at 65% of the diet dry matter (DM). The results showed that straw pH reduced from 11.58 to 8.60 after 6 weeks of ensilage. The ET diet increased average DM digestibility and intake by 19% and 43% respectively. Total water intake was similar across treatments, while the water/DM intake ratio was 23% higher with the UT diet than with the ET one. Ruminal (6.73 vs. 6.84) and faecal (8.67 vs. 9.05) pH decreased but urinary pH (6.14 vs. 8.13) increased as a result of feeding animals on the ET diet compared with the UT diet. Compared with the UT diet, the ET one decreased faecal fibre (12%), moisture (32%) and water holding capacity, while it increased faecal ash (10%) and density (20%). The volume of urine excreted by the sheep fed with the ET diet increased by 67%, but their urine specific gravity (SG) decreased. No significant effects were observed for the dietary N supplementation and interactions between straw type × N supplementation with regard to any of the measured characteristics except for DM intake, which reduced due to the casein supplementation in the ET diet. These results indicate that the alkali treatment and ensilage of barley straw increased digestibility, intake, faecal consistency and urinary pH and dilution but decreased straw alkalinity as well as ruminal and faecal pH.

Effects of sheep and mouse urine on the growth pattern of Leishmania major promastigotes.

The protozoan parasites of the genus Leishmania are the causative agents of different clinical diseases. Fetal calf serum (FCS) is the main part and the most expensive ingredient of the Leishmania culture media. Here, the efficacies of different concentrations (1%, 2.5%, 5%, and 10%) of the filtered and autoclaved sheep and mouse urine were evaluated as a growth stimulator in Leishmania culture procedure. The results indicated that culture media enriched with the filtered sheep and mouse urine supported the growth of the parasites and can be used for cultivation of Leishmania parasites. In conclusion, this study has demonstrated an alternative low-cost medium that could be used in cultivation process of Leishmania major promastigotes.

Regulation of intramembranous absorption and amniotic fluid volume by constituents in fetal sheep urine.

Our objective was to test the hypothesis that fetal urine contains a substance(s) that regulates amniotic fluid volume by altering the rate of intramembranous absorption of amniotic fluid. In late gestation ovine fetuses, amniotic fluid volumes, urine, and lung liquid production rates, swallowed volumes and intramembranous volume and solute absorption rates were measured over 2-day periods under control conditions and when urine was removed and continuously replaced at an equal rate with exogenous fluid. Intramembranous volume absorption rate decreased by 40% when urine was replaced with lactated Ringer solution or lactated Ringer solution diluted 50% with water. Amniotic fluid volume doubled under both conditions. Analysis of the intramembranous sodium and chloride fluxes suggests that the active but not passive component of intramembranous volume absorption was altered by urine replacement, whereas both active and passive components of solute fluxes were altered. We conclude that fetal urine contains an unidentified substance(s) that stimulates active intramembranous transport of amniotic fluid across the amnion into the underlying fetal vasculature and thereby functions as a regulator of amniotic fluid volume.

Excretion of loline alkaloids in urine and faeces of sheep dosed with meadow fescue (Festuca pratensis) seed containing high concentrations of loline alkaloids.

AIM: To determine the effect of oral dosing of sheep with loline alkaloids on their excretion in urine and faeces, and to monitor for any toxic effects. METHODS: In Experiment 1, six 9-month-old ewe lambs were given a single oral dose of loline alkaloids (52 mg/kg bodyweight (BW); acute exposure) as a suspension of ground meadow fescue (Festuca pratensis) seed in water. In Experiment 2, on six consecutive days, six ewe lambs were given three doses of loline (68 mg/kg BW/day; chronic exposure). Blood was collected at variable intervals up to 72 h in Experiment 1, and up to 8 days in Experiment 2, for haematology and measurement of alkaline phosphatase, aspartate aminotransaminase, creatine kinase and γ-glutamyl transferase in plasma. Urine and faecal samples were collected at similar times for measurement of creatinine in urine and loline alkaloid analysis. A post mortem with histopathology was carried out on two animals at the end of each experiment. RESULTS: The loline alkaloids, N-acetyl norloline, N-formyl loline, N-acetyl loline, N-methyl loline and loline base were detected in urine within 15 minutes after the single dosing. N-formyl loline and loline base were the predominant metabolites in urine in both experiments. The total quantity of lolines excreted in both urine and faeces was 10% and 4% of the amount dosed in Experiments 1 and 2, respectively. In both experiments, the clinical chemistry parameters in blood and urine were within normal ranges. Post-mortem and histopathological examination did not show any abnormalities. CONCLUSIONS: This is the first report of loline alkaloid profiles in both urine and faeces of sheep. The appearance of loline alkaloids and the loline base in urine within 15 minutes suggests rapid uptake, metabolism and excretion. Loline alkaloids were non-toxic to sheep at the concentrations they are exposed to under New Zealand grazing conditions. The low recovery of loline alkaloids in urine and faeces in the absence of toxicity signs suggests lolines are extensively metabolised; probably to forms other than N-formyl loline, N-methyl loline, N-acetyl loline, N-acetyl norloline, and loline base in the digestive tract of sheep prior to absorption, and/or in the liver or other tissues following absorption.

Laboratory diagnostics in the urine of young and pregnant ewes.

OBJECTIVE: The purpose of this study was to identify values for net acid base excretion (NABE) which are significant indicators of the acid-base equilibrium in pregnant and young ewes and to show its relationship with other parameters (base, acid, ammonium [NH4], base-acid quotient, sodium [Na], potassium [K], calcium [Ca]) in ovine urine. In contrast to dairy cows, data are rare on these parameters in ewes. MATERIAL AND METHODS: A total of 99 animals were used in the study, consisting of 56 young (average of 5.6±1.1 months) and 43 pregnant ewes (average of 35.2±18.8 months). Measurement of fractional NABE in urine samples was carried out according to the method reported by Kutas. The pH value of the urine was measured with a laboratory pH meter. Na, K and total Ca were measured with a flame photometer. RESULTS: For all values except Na significant differences occurred between urine samples of pregnant ewes and young ewes (p<0.001). Base, acid, NH4, NABE, K and Ca values were significantly higher in the urine of the youngs than in pregnant ewes. In young ewes, a strong correlation was found between NABE and base values while a weak correlation could be observed between pH and base values. In pregnant ewes, strong NABE-base, NABE-K, K-acid and K-base correlations were found as well as weak NH4-base, NH4-NABE and NH4-K correlations. There was a strongly positive correlation between NABE and NH4 in pregnant ewes, while a weak negative correlation between those values was observed in young ewes. CONCLUSION: For the first time, we established values for NABE and certain other parameters in urine of pregnant ewes and young ewes. It was shown that the acid-base balance in pregnant ewes and young ewes can be evaluated by measuring NABE and certain trace elements in urine like in cattle.

Haemodynamics of lambs grazing perennial ryegrass (Lolium perenne L.) either infected with AR6 novel, wild-type endophyte, or endophyte-free.

AIM: To compare vasoconstriction of the auricular artery and characteristics of blood flow in the carotid arteries between lambs grazing perennial ryegrass that was either infected with the AR6 novel endophyte (AR6), wild-type endophyte or endophyte-free. METHODS: Sixteen Coopworth lambs, mean 23.7 (SD 1.8) kg, were randomly assigned to graze three, 0.10-ha pastures of perennial ryegrass (cultivar Extreme) located in Lincoln, New Zealand, that were infected with either the AR6 novel endophyte (n=5) or wild-type endophyte (n=6) or were endophyte-free (n=5), for 18 days until 16 March 2009. Lambs on AR6 pasture were then switched to endophyte-free pasture, and those on endophyte-free were switched to AR6 pasture, for 18 days. Lambs continued grazing the wild-type ryegrass during both phases of the study. Colour Doppler ultrasonography was used to monitor cross-sectional area of lumina in the auricular and carotid arteries as measurements of vasoconstriction, and to measure pulsatility indices, heart rate, systolic and diastolic velocities, and mean velocity in the carotid artery. Urine was sampled once during each phase, to measure the concentration of urinary alkaloids. A heat challenge (32°C) was imposed on the last day of the experiment, to determine treatment effects on rectal temperature and respiration rate. RESULTS: Vasoconstriction was detected in the auricular arteries of lambs grazing AR6 and wild-type pastures. Areas of lumina declined linearly over 9 days in lambs that were switched from endophyte-free to AR6 pastures (p<0.05), and areas increased linearly in lambs that were switched from AR6 to endophyte-free pasture (p<0.001). Resistance to blood flow in the carotid arteries decreased linearly in the 9 days after lambs were switched from AR6 to endophyte-free pasture(p<0.05), and tended to increase after lambs on endophyte-free pasture were switched to AR6 (p<0.10). The concentrations of urinary alkaloids decreased after switching lambs from AR6 to endophyte-free pasture, and increased after switching from endophyte-free to AR6 pasture (p<0.05). The concentrations of urinary alkaloids of lambs on wild-type pasture were similar between the conditioning and experimental phases. There were no treatment effects on rectal temperature and respiration rate during the heat challenge. CONCLUSIONS: These results indicated that management approaches are needed to overcome potential vulnerabilities to heat stress for sheep grazing AR6 perennial ryegrass. Furthermore, following grazing such pasture, lambs will need to graze endophyte-free ryegrass for >18 days, to completely clear ergot alkaloids from their vasculature, assuming that complete clearance can be achieved.

Prenatal glucocorticoid exposure in the sheep alters renal development in utero: implications for adult renal function and blood pressure control.

Treatment of the pregnant ewe with glucocorticoids early in pregnancy results in offspring with hypertension. This study examined whether glucocorticoids can reduce nephron formation or alter gene expression for sodium channels in the late gestation fetus. Sodium channel expression was also examined in 2-mo-old lambs, while arterial pressure and renal function was examined in adult female offspring before and during 6 wk of increased dietary salt intake. Pregnant ewes were treated with saline (SAL), dexamethasone (DEX; 0.48 mg/h) or cortisol (CORT; 5 mg/h) over days 26-28 of gestation (term = 150 days). At 140 days of gestation, glomerular number in CORT and DEX animals was 40 and 25% less, respectively, compared with SAL controls. Real-time PCR showed greater gene expression for the epithelial sodium channel (α-, ß-, γ-subunits) and Na(+)-K(+)-ATPase (α-, ß-, γ-subunits) in both the DEX and CORT group fetal kidneys compared with the SAL group with some of these changes persisting in 2-mo-old female offspring. In adulthood, sheep treated with dexamethasone or cortisol in utero had elevated arterial pressure and an apparent increase in single nephron glomerular filtration rate, but global renal hemodynamics and excretory function were normal and arterial pressure was not salt sensitive. Our findings show that the nephron-deficit in sheep exposed to glucocorticoids in utero is acquired before birth, so it is a potential cause, rather than a consequence, of their elevated arterial pressure in adulthood. Upregulation of sodium channels in these animals could provide a mechanistic link to sustained increases in arterial pressure in cortisol- and dexamethasone-exposed sheep, since it would be expected to promote salt and water retention during the postnatal period.

Quantification of melamine absorption, distribution to tissues, and excretion by sheep.

Eight Döhne Merino rams were used to quantify apparent absorption, distribution to tissues, and excretion of dietary melamine in sheep. Two batches of concentrate pellets were made; one (CON) contained corn gluten meal with no detectable melamine and the other (MEL) contained corn gluten meal that was previously found to be highly contaminated with melamine at 15,117 mg/kg. The MEL pellets contained 1,149 mg/kg of melamine. During a 10-d adaptation period, all the animals received a forage-based diet supplemented with 600 g/d of the CON pellets. This was followed by an 8-d collection period during which 6 of the animals received MEL pellets and 2 received CON pellets. Melamine intake of sheep that received MEL pellets was 0.69 g/d. Blood samples were taken before first ingestion of MEL pellets on d 1 and again on d 3, 6, and 8 of the collection period for melamine and serum creatinine analyses. Feces and urine were collected quantitatively over the 8 d for proximate and melamine analyses. All the animals were slaughtered at the end of the trial, and samples of the LM, liver, kidneys, and abdominal fat were taken for melamine analysis. Data of the 2 sheep that received CON pellets for the duration of the trial confirmed that no melamine was detected in any of the samples, and no statistical analyses were performed on these data. The apparent digestibility or efficiency of absorption of ingested melamine was 76.7%. Melamine was detected in the urine, blood, muscle (LM), and fat tissue of all the sheep that received MEL pellets. Serum melamine concentrations reached 5.4 mg/kg on d 8 of the collection period, and the meat (LM) contained 9.6 mg/kg of melamine. Calculations on the partitioning of ingested melamine suggested that urine is the major excretion route accounting for 53.2%, whereas feces accounted for 23.3% of ingested melamine. Approximately 3.5% of the ingested melamine was detected in muscle. It was concluded that ingested melamine is highly absorbable from the small intestine and that a pathway exists for the distribution of dietary melamine to meat.

Effect of dietary cation-anion difference on urinary pH, feedlot performance, nitrogen mass balance, and manure pH in open feedlot pens.

Six experiments were conducted to evaluate dietary cation-anion difference (DCAD) in concentrate diets on urinary pH, feedlot performance, and N mass balance. In Exp. 1, 15 wether lambs (33.5 ± 3.0 kg) in five 3 × 3 Latin squares were fed a basal diet of 82.5% dry-rolled corn (DRC), 7.5% alfalfa hay, 5% molasses, and 5% supplement with different proportions of anionic and cationic salts. The DCAD was -45, -24, -16, -8, 0, +8, +16, +24, +32, and +40 mEq per 100 g of DM with the control basal diet (DCAD = +8) included in each square. Urinary pH increased (cubic, P < 0.01) as DCAD increased and DMI increased linearly (P < 0.01) with increasing DCAD. In Exp. 2 and 3, 8 Holstein steers (312 ± 24 kg) were used in 2 consecutive 4 × 4 Latin squares. Steers were fed either the same basal diet as Exp. 1 or a basal diet with 20% wet distillers grains (WDGS) replacing DRC. In Exp. 2, DCAD was adjusted to -2, -12, and -22 mEq per 100 g of DM from the basal diet (DCAD = +8) and DCAD was adjusted in Exp. 3 to -12, -22, and -32 mEq per 100 g of DM from the basal WDGS diet (DCAD = -2). Urinary pH decreased linearly as DCAD decreased (P < 0.01) in both experiments, whereas DMI decreased linearly in Exp. 2 (P = 0.02) but not Exp. 3 (P = 0.96). In Exp. 4, 6 crossbred steers (373 ± 37 kg) were used in a 2-period crossover design. Steers were fed the same basal diet as Exp. 3 with DCAD of -16 (NEG) and +20 (POS) mEq per 100 g of DM. Urinary pH and DMI (P < 0.05) were less for cattle fed the NEG diet compared with POS. In 2 experiments, steers (n = 96 each) were fed NEG or POS as calves (260 ± 22 kg of BW) for 196 d from November to May (Exp. 5) or as yearlings (339 ± 32 kg of BW) for 145 d from June to October (Exp. 6). Final BW, DMI, ADG, and HCW were not different (P > 0.11) among treatments in either experiment. Efficiency of BW gain was increased (P = 0.05) for steers fed NEG compared with POS in Exp. 5 but was not different (P = 0.11) in Exp. 6. Amount of N intake, retention, excretion, and manure N (kg/steer) were not different (P > 0.11) among treatments in either experiment. Manure pH (soil, feces, and urine) was decreased (P < 0.01) in pens fed NEG compared with POS in both experiments. Amount of N lost (kg/steer) was not different (P = 0.44) in Exp. 5, but tended (P = 0.09) to be 10.6% greater for POS compared with NEG in Exp. 6. Urinary pH was decreased by reducing DCAD, but this had minimal effect on N losses in open feedlot pens in these experiments.

Response of fungal, bacterial and ureolytic communities to synthetic sheep urine deposition in a grassland soil.

In grazed pastures, soil pH is raised in urine patches, causing dissolution of organic carbon and increased ammonium and nitrate concentrations, with potential effects on the structure and functioning of soil microbial communities. Here we examined the effects of synthetic sheep urine (SU) in a field study on dominant soil bacterial and fungal communities associated with bulk soil and plant roots (rhizoplane), using culture-independent methods and a new approach to investigate the ureolytic community. A differential response of bacteria and fungal communities to SU treatment was observed. The bacterial community showed a clear shift in composition after SU treatment, which was more pronounced in bulk soil than on the rhizoplane. The fungal community did not respond to SU treatment; instead, it was more affected by the time of sampling. Redundancy analysis of data indicated that the variation in the bacterial community was related to change in soil pH, while fungal community was more responsive to dissolution of organic carbon. Like the universal bacterial community, the ureolytic community was influenced by the SU treatment. However, different taxa within the ureolytic bacterial community responded differentially to the treatment. The ureolytic community comprised of members from a range of phylogenetically different taxa and could be used to measure the effect of environmental perturbations on the functional diversity of natural ecosystems.

Pharmacokinetics and milk distribution characteristics of orbifloxacin following intravenous and intramuscular injection in lactating ewes.

The purpose of the current investigation is to elucidate the pharmacokinetic profiles of orbifloxacin (OBFX) in lactating ewes (n = 6) following intravenous (i.v.) and intramuscular (i.m.) administrations of 2.5 mg/kg W. In a crossover study, frequent blood, milk, and urine samples were drawn for up to 48 h after the end of administration, and were then assayed to determine their respective drug concentrations through microbiological assay using Klebsiella pneumoniae as the test micro-organism. Plasma pharmacokinetic parameters were derived from plasma concentration-time data using a compartmental and noncompartmental analysis, and validated a relatively rapid elimination from the blood compartment, with a slope of the terminal phase of 0.21 +/- 0.02 and 0.19 +/- 0.06 per hour and a half-life of 3.16 +/- 0.43 and 3.84 +/- 0.59 h, for i.v. and i.m. dosing, respectively. OBFX was widely distributed with a volume of distribution V((d(ss))) of 1.31 +/- 0.12 L/kg, as suggested by the low percentage of protein binding (22.5%). The systemic body clearance (Cl(B)) was 0.32 +/- 0.12 L/h x kg. Following i.m. administration, the maximum plasma concentration (C(max)) of 1.53 +/- 0.34 microg/mL was reached at t(max) 1.25 +/- 0.21 h. The drug was completely absorbed after i.m. administration, with a bioavailability of 114.63 +/- 11.39%. The kinetic milk AUC(milk)/AUC(plasma) ratio indicated a wide penetration of orbifloxacin from the bloodstream to the mammary gland. OBFX urine concentrations were higher than the concurrent plasma concentrations, and were detected up to 30 h postinjection by both routes. Taken together, these findings indicate that systemic administration of orbifloxacin could be efficacious against susceptible mammary and urinary pathogens in lactating ewes.

Pharmacokinetics of monepantel and its sulfone metabolite, monepantel sulfone, after intravenous and oral administration in sheep.

The pharmacokinetic properties of the developmental Amino-Acetonitrile Derivative (AAD), monepantel and its sulfone metabolite, monepantel sulfone were investigated in sheep following intravenous (i.v.) and oral administrations. The sulfone metabolite was rapidly formed and predominated over monepantel 4 h after dosing, irrespective of the route of administration. The steady-state volume of distribution, total body clearance and mean residence time of monepantel were 7.4 L/kg, 1.49 L/(kg x h) and 4.9 h, respectively and 31.2 L/kg, 0.28 L/(kg x h) and 111 h, respectively for monepantel sulfone. The overall bioavailability of monepantel was 31%, but it was demonstrated that approximately the same amount of monepantel sulfone was produced whether monepantel was given intravenously or orally (AUC((0-infinity)) oral/AUC((0-infinity)) i.v. of 94% for monepantel sulfone), making oral administration a very efficient route of administration for monepantel in terms of the amount of sulfone metabolite generated. Because monepantel sulfone is the main chemical entity present in sheep blood after monepantel administration and because it is also an active metabolite, its pharmacokinetic properties are of primary importance for the interpretation of future residue and efficacy studies. Overall, these pharmacokinetic data aid in the evaluation of monepantel as an oral anthelmintic in sheep.

Urolitíase em caprinos e ovinos / Urolithiasis in sheep and goats

A urolitíase é uma doença freqüente no semi-árido em ovinos e caprinos alimentados com grãos, principalmente quando a relação Ca:P não é corrigida e são utilizadas grandes quantidades de concentrado. De 28 ovinos com urolitíase atendidos no Hospital Veterinário da UFCG, 19 morreram e 9 receberam alta, dos 15 caprinos 12 morreram e 3 receberam alta. As alterações mais freqüentes foi presença de cálculos na uretra e bexiga, uretrite hemorrágica difusa severa, uretrite purulenta, ruptura de uretra com presença de urina no tecido subcutâneo, cistite hemorrágica necrosante, ruptura de bexiga, hidronefrose, nefrite necrosante difusa, abscesso renal, pielonefrite, hemorragia renal e ruptura renal. A gravidade das lesões observadas justifica a alta letalidade apesar dos tratamentos medicamentosos e cirúrgicos. A medida preventiva mais importante é a correção dos níveis de Ca e P. Recomenda-se, também, a administração de volumosos em boa quantidade e qualidade e, nos casos em que a quantidade de concentrados seja superior a 1,5 por cento do peso vivo, a administração de cloreto de amônia a 1 por cento no concentrado e a administração de cloreto de sódio em concentrações de 0,5 até 4 por cento. Deve ser administrada água de boa qualidade à vontade. É necessário abolir a prática, extremamente utilizada em caprinos e ovinos, de administrar sal mineral em animais alimentados com grãos e subprodutos dos mesmos.

Urolithiasis is a common disease in the Brazilian semiarid in rams and bucks fed with grains, mainly with low C:P ratio. From 28 rams with urolithiasis sent to the Veterinary Hospital of the Federal University of Campina Grande, Paraíba, 19 died and 9 survived, and from the bucks, 13 died and 3 survived At necropsy the main lesions were presence of calculi in the urethra and urinary bladder, diffuse purulent urethritis, hemorrhagic necrotizing urethritis, rupture of the urethra with presence of urine in the subcutaneous tissue, hydronephrosis, urinary bladder rupture, necrotizing diffuse nephritis, pyelonephritis, renal hemorrhage, and renal rupture. The severity of the lesions is responsible for the high case fatality rate despite medical and chirurgic treatment. The most important preventive measure is the correction of the Ca:P ratio to at least 2:1. The administration of good quality roughage in the food is also necessary. In cases in which the amount of concentrate food is higher than 1.5 percent live weight, the addition of 1 percent ammonium chloride and 0.5-4 percent of sodium chloride in the diet could be also necessary. Water had to be offered ad libitum. It is necessary to abolish the use of ad libitum mineral supplementation in sheep and goats fed grains or their byproducts.

Tissue residues and urinary excretion of zilpaterol in sheep treated for 10 days with dietary zilpaterol.

Zilpaterol is a beta-adrenergic growth promoter approved in Mexico and South Africa for use in cattle. Understanding the rates of zilpaterol depletion from tissues and urine is of interest for the development of strategies to detect the off-label use of zilpaterol. Eight sheep were fed 0.15 mg/kg/day dietary zilpaterol hydrochloride (Zilmax) for 10 consecutive days; two sheep each were slaughtered 0, 2, 5, and 9 days after discontinuation of exposure to the zilpaterol-containing diet. Tissue zilpaterol levels rapidly decreased during the withdrawal period. On the basis of LC-MS/MS-ES (external standard) measurements, liver zilpaterol residues in sheep were 29.3, 1.5, 0.13, and 0.10 ng/g after 0, 2, 5, and 9 day withdrawal periods, respectively; kidney residues were 29.6, 1.10, and 0.09 ng/g and below the detection limit; and muscle residues were 13.3, 0.86, 0.12, and 0.08 ng/g at the same respective withdrawal periods. Between-animal variation in urinary zilpaterol concentrations during the feeding period was considerable, although zilpaterol concentrations converged somewhat as steady state was reached. During the first 3 days of the withdrawal period, zilpaterol elimination followed a first-order excretion pattern, having an average elimination half-life of 15.3 +/- 1.8 h. Urinary zilpaterol concentrations during the withdrawal period were determined using ELISA, HPLC-fluorescence, LC-MS/MS-ES (external standard), and LC-MS/MS-IS (internal standard). Comparison of these methods showed a high correlation with each other. With the exception of LC-MS/MS-IS, the regression coefficients of the linear equations with a zero intercept were between 0.90 and 1.25, indicating the near equivalence of the methods. Because of its simplicity, ELISA is a convenient assay for determining zilpaterol levels in urine giving similar results to HPLC-fluorescence and LC-MS/MS-ES without requiring the extensive cleanup of the latter methods.