Evaluation of three herbal compounds used for the management of lower urinary tract disease in healthy cats: a pilot study.

OBJECTIVES: Lower urinary tract disease (LUTD) occurs commonly in cats, and idiopathic cystitis (FIC) and urolithiasis account for >80% of cases in cats <10 years of age. Although several strategies have been recommended, a common recommendation is to induce dilute urine resulting in more frequent urination and to dilute calculogenic constituents. In addition to conventional therapy using modified diets, traditional Chinese and Western herbs have been recommended, although only one - choreito - has published data available. We evaluated three commonly used herbal treatments recommended for use in cats with LUTD: San Ren Tang, Wei Ling Tang and Alisma. We hypothesized that these three Chinese herbal preparations would induce increased urine volume, decreased urine saturation for calcium oxalate and struvite, and differences in mineral and electrolyte excretions in healthy cats. METHODS: Six healthy spayed female adult cats were evaluated in a placebo-controlled, randomized, crossover design study. Cats were randomized to one of four treatments, including placebo, San Ren Tang, Wei Ling Tang or Alisma. Treatment was for 2 weeks each with a 1 week washout period between treatments. At the end of each treatment period, a 24 h urine sample was collected using modified litter boxes. RESULTS: Body weights were not different between treatments. No differences were found in 24 h urinary analyte excretions, urine volume, urine pH or urinary saturation for calcium oxalate or struvite between treatments. CONCLUSIONS AND RELEVANCE: The results of this study do not support the hypothesis; however, evaluation of longer-term and different dosage studies in cats with LUTD is warranted.

Effect of magnesium dose on amount of pharmaceuticals in struvite recovered from urine.

Phosphorus (P) recovery was carried out through struvite precipitation from urines. Human urine, however, contains not only high nutrients for plants, such as P and nitrogen, but also pharmaceuticals and hormones. In this work, effects of magnesium (Mg) dose (in terms of Mg:P ratio) on P recovery efficiency and pharmaceutical amounts contained in struvite were investigated. Batch-scale experiments of synthetic and human urines revealed that struvite precipitation formed more X-shaped crystals with an increased molar ratio of Mg:P, while the amount of pharmaceuticals (tetracycline, demeclocycline, and oxytetracycline) in struvite decreased with an increased molar ratio of Mg:P. The lowest pharmaceutical amounts in struvite were found at the Mg:P ratio of 2:1 from both samples. Moreover, the maximum P recovery efficiency, quantity and purity of struvite were found in the range of 1.21 to 2:1. It indicated that the molar ratio of Mg:P has a significant impact on struvite precipitation in terms of pharmaceutical amounts in struvite; morphology, quantity and purity of struvite; and P recovery.

Marked struvite crystalluria and its association with lower urinary tract signs in a cat with feline idiopathic cystitis.

CASE REPORT: We describe a case of a large amount of mineralised material, presumed to be struvite crystals, within the urinary bladder of a cat with feline idiopathic cystitis. The presence of this material coincided with episodes of lower urinary tract signs in this cat over a 2-year period. CLINICAL SIGNIFICANCE: Although struvite crystalluria is widely considered to be clinically insignificant, this generalisation may not be true for all cats with lower urinary tract disease. Imaging of the urinary tract is recommended in all cats with lower urinary tract signs.

Effects of two calculolytic diets on parameters of feline mineral metabolism.

OBJECTIVES: To evaluate the influence of two feline calculolytic diets on selected parameters of mineral metabolism. MATERIALS AND METHODS: Two dry commercial diets designed for struvite urolith dissolution were evaluated in 14 cats. The study was designed as a two-sequence, four-period crossover protocol with a baseline period, two 60-day "run-in" periods in which calculolytic diets (Diet 1 and Diet 2) were fed and one 30-day "wash-out" period. Data are expressed as median (range). RESULTS: Feeding the calculolytic diets for two months did not alter plasma concentrations of calcium, phosphorus, magnesium and parathyroid hormone. A significant (P < 0.05 in each case) decline in calcitriol was observed after administering both diets from 236.4 (122.4-429.6) to 170.4 (108.0-394.3) pmol/L (Diet 1) and from 278.4 (153.6-492.0) to 177.1 (87.6-392.4) pmol/L (Diet 2). Cats fed Diet 1 showed a significant increase in urine calcium concentration (from 0.3 (0.2-0.5) to 0.4 (0.3-0.7) mmol/L). Magnesium concentration in urine was significantly increased with both diets, from 1.4 (0.1-1.7) to 1.5 (1.3-2.4) mmol/L (Diet 1) and from 1.1 (0.4-1.9) to 2.0 (0.1-3.1) mmol/L (Diet 2). CLINICAL SIGNIFICANCE: Both diets resulted in an increased urinary concentration of magnesium, through different mechanisms: urine acidification (Diet 1) and increased sodium load (Diet 2).

Life cycle comparison of centralized wastewater treatment and urine source separation with struvite precipitation: Focus on urine nutrient management.

Alternative approaches to wastewater management including urine source separation have the potential to simultaneously improve multiple aspects of wastewater treatment, including reduced use of potable water for waste conveyance and improved contaminant removal, especially nutrients. In order to pursue such radical changes, system-level evaluations of urine source separation in community contexts are required. The focus of this life cycle assessment (LCA) is managing nutrients from urine produced in a residential setting with urine source separation and struvite precipitation, as compared with a centralized wastewater treatment approach. The life cycle impacts evaluated in this study pertain to construction of the urine source separation system and operation of drinking water treatment, decentralized urine treatment, and centralized wastewater treatment. System boundaries include fertilizer offsets resulting from the production of urine based struvite fertilizer. As calculated by the Tool for the Reduction and Assessment of Chemical and Other Environmental Impacts (TRACI), urine source separation with MgO addition for subsequent struvite precipitation with high P recovery (Scenario B) has the smallest environmental cost relative to existing centralized wastewater treatment (Scenario A) and urine source separation with MgO and Na3PO4 addition for subsequent struvite precipitation with concurrent high P and N recovery (Scenario C). Preliminary economic evaluations show that the three urine management scenarios are relatively equal on a monetary basis (<13% difference). The impacts of each urine management scenario are most sensitive to the assumed urine composition, the selected urine storage time, and the assumed electricity required to treat influent urine and toilet water used to convey urine at the centralized wastewater treatment plant. The importance of full nutrient recovery from urine in combination with the substantial chemical inputs required for N recovery via struvite precipitation indicate the need for alternative methods of N recovery.

Chronic trimethyltin chloride exposure and the development of kidney stones in rats.

We recently reported that occupational exposure to trimethyltin (TMT) is a risk factor for developing kidney stones. To further examine the association between TMT exposure and the formation of kidney stones, we conducted a 180-day animal study and exposed the randomly grouped Sprague-Dawley (SD) rats to TMT in the drinking water at doses of 0, 8.2, 32.8 and 131.3 µg kg(-1) day(-1). Transient behavioral changes were observed in the high-dose group during the first 2 weeks of exposure. TMT exposure led to a significant dose-dependent inhibition of renal H(+)/K(+)-ATPase and an increase in urinary pH. In comparison to no kidney stones being identified in the control and the lowest dose group, 1 rat in the 32.8 µg kg(-1) day(-1) dose group and 3 out of 9 rats in the 131.3 µg kg(-1) day(-1) dose group were found to have stones in the kidney/urinary tract. Pathological analysis showed that more wide spread calcium disposition was observed in kidneys of rats with TMT exposure compared with the rats in the control group. However, X-ray diffraction (XRD) analysis found that the kidney stones were mainly composed of struvite with the formula: NH4MgPO4 6H2O, while calcium-containing components were also detected. Together, this study further demonstrates through animal studies that chronic exposure to a relatively low level of TMT induces nephrotoxicity and increases the risk for developing kidney stones.

Phosphorus recovery: minimization of amount of pharmaceuticals and improvement of purity in struvite recovered from hydrolysed urine.

Struvite (MgNH4PO4·6H2O) is normally used as a fertilizer in agriculture, where struvite crystallization from hydrolysed human urine is a simple and reliable method for phosphorus (P) recovery. Human urine, however, contains high amount of pharmaceuticals, which may cause health risk for applications. This research investigates the possibility of decreasing the amount of pharmaceuticals (tetracycline, demeclocycline and oxytetracycline) in struvite crystals recovered from synthetic and human urines by focusing on storage time, and of increasing the quality of struvite production. Urines were stored for different times up to 15 days prior to recovery of phosphorus by two steps, spontaneous precipitation and struvite crystallization. The morphology of spontaneous precipitates and struvite crystals was observed. Spontaneous precipitation removed around 17-24% of phosphate from synthetic and human urines, while pharmaceuticals were removed with a quite high amount at a short storage time (5 days) and this amount decreased with increasing the storage time (10 and 15 days). Urines with>70% remaining phosphates were re-used for struvite crystallization by adding extra magnesium. It was found that maximum P-recovery efficiency could be achieved from struvite crystallization at 5-day storage time, 70% and 68% of remaining P in the separated supernatant from synthetic and human urines, respectively, whereas less than 1% pharmaceuticals remained in the struvite crystals from both samples. This indicates that the procedure in this work is a good method for phosphorus recovery, in which high struvite purity (>99%) is obtained with low amount of pharmaceuticals.

[New diagnostic tool for individual monitoring of urolithiasis risk and other metabolic diseases]. / Neues Harn-Diagnostikverfahren zur individuellen Überwachung des Harnsteinrisikos und anderer Stoffwechselerkrankungen.

Urinary composition is the result of the interplay of all metabolic processes, including all metabolites and toxins produced. Any change in urine composition influences urinary supersaturation, the major thermodynamic driving force of crystal formation. Urolithiasis is a wide-spread disease with a prevalence rate of 4% to 10%. Formation risk of calcium-oxalate, by far the most common mineral in uroliths, is quantified by the Bonn-Risk-Index (BRI). The BRI measures induced crystal growth within native urine and shows superior diagnostic sensitivity and specificity compared to other urolithiasis risk indices. The concept of BRI quick test presented in this work allowes even untrained persons to easily determine the urolithiasis risk. Many diseases lack a simple, easily accessible and cost effective diagnostic approach to monitor their course and treatment success. Since BRI takes into account every constituent of native urine, it can be used to monitor a wide range of metabolic diseases.

In vitro studies on the role of glycosaminoglycans in crystallization intensity during infectious urinary stones formation.

Proteus mirabilis cause urinary tract infections which are recurrent and can lead to formation of urinary calculi. Both bacterial and the host factors are involved in the development of urolithiasis. To determine the impact of glycosaminoglycans (GAGs) in the formation of P. mirabilis-induced urinary stones, we investigated the in vitro crystallization, aggregation and adhesion of crystals in the presence of GAGs naturally appearing in urine. Crystallization experiments were performed in synthetic urine infected with P. mirabilis in the presence of: hyaluronic acid (HA), heparan sulfate (HS), chondroitin sulfate A, B and C (ChSA, ChSB, ChSC). The intensity of crystallization and aggregation were established by counting particles and phase-contrast microscopy. To analyze the adhesion of crystals, we used normal urothelium and (45)Ca isotope-labeled crystals. In the presence of ChSC, both the size of the crystals formed and their number were higher compared with the control. GAGs increased crystals adhesion to the cells, but only for ChSA this effect was significant. Chondroitin sulfates, which accelerate the first stages of infection-induced stones formation, may play an important role in the pathogenesis of infectious urolithiasis.

Efficacy of two commercially available, low-magnesium, urine-acidifying dry foods for the dissolution of struvite uroliths in cats.

OBJECTIVE: To compare the efficacy and safety of using 2 commercially available, low-magnesium, urine-acidifying dry foods to dissolve sterile struvite uroliths in cats. DESIGN: Prospective, multicenter, randomized clinical trial . SAMPLE: 37 cats with presumed struvite uroliths. PROCEDURES: Cats were randomly assigned to be fed 1 of 2 low-magnesium, urine-acidifying dry foods (food A or B). For each cat, physical examination, urinalysis, and abdominal radiography were performed weekly to assess treatment response. RESULTS: 32 cats had complete urolith dissolution. Mean ± SD times for a 50% reduction in urolith size (0.69 ± 0.1 weeks) and complete urolith dissolution (13.0 ± 2.6 days) were significantly shorter for cats fed food A, compared with those (1.75 ± 0.27 weeks and 27.0 ± 2.6 days, respectively) for cats fed food B. At study termination, mean ± SD urine pH (6.083 ± 0.105) for cats fed food A was lower than that (6.431 ± 0.109) for cats fed food B. In 5 cats, uroliths did not dissolve and were subsequently determined to be composed of 100% ammonium urate (n = 4) or 100% calcium oxalate (1). Adverse events associated with diet were not observed in any of the cats. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that dietary dissolution is safe and effective for eradication of sterile struvite uroliths in cats. Cats fed food A had faster urolith dissolution than did cats fed food B. Lack of a reduction in urolith size at 2 weeks after diet initiation was indicative of misdiagnosis or noncompliance.

Influence of acidifying or alkalinizing diets on bone mineral density and urine relative supersaturation with calcium oxalate and struvite in healthy cats.

OBJECTIVE: To evaluate the influence of acidifying or alkalinizing diets on bone mineral density and urine relative supersaturation (URSS) with calcium oxalate and struvite in healthy cats. ANIMALS: 6 castrated male and 6 spayed female cats. PROCEDURES: 3 groups of 4 cats each were fed diets for 12 months that differed only in acidifying or alkalinizing properties (alkalinizing, neutral, and acidifying). Body composition was estimated by use of dual energy x-ray absorptiometry, and 48-hour urine samples were collected for URSS determination. RESULTS: Urine pH differed significantly among diet groups, with the lowest urine pH values in the acidifying diet group and the highest values in the alkalinizing diet group. Differences were not observed in other variables except urinary ammonia excretion, which was significantly higher in the neutral diet group. Calcium oxalate URSS was highest in the acidifying diet group and lowest in the alkalinizing diet group; struvite URSS was not different among groups. Diet was not significantly associated with bone mineral content or density. CONCLUSIONS AND CLINICAL RELEVANCE: Urinary undersaturation with calcium oxalate was achieved by inducing alkaluria. Feeding an alkalinizing diet was not associated with URSS with struvite. Bone mineral density and calcium content were not adversely affected by diet; therefore, release of calcium from bone caused by feeding an acidifying diet may not occur in healthy cats.

Urinary turbidity as a marker of crystallization: is spectrophotometric assessment useful?

PURPOSE: We propose a simple and inexpensive in vitro crystallization assay of measuring turbidity by spectrophotometry in synthetic urine. We validated our method by investigating the effect of potassium (K) citrate on the crystallization of calcium oxalate monohydrate (CaOx), calcium phosphate, and magnesium ammonium phosphate using synthetic urine. METHODS: The crystallization of CaOx was studied using turbidimetric measurements of solution produced by mixing calcium chloride and sodium oxalate at 37 °C, pH 5.7. The turbidity of the crystal suspension was measured immediately with double-beam spectrophotometer as the absorbance of light at 660 nm wavelength. The rates of crystal formation and aggregation were obtained by measuring optical density (OD) over 30 min. The obtained results were compared to CaOx crystal concentration with and without citrate assessed by optical microscopy. RESULTS: The sensitivity of spectrophotometry in measuring turbidity was confirmed by the linear correlation between the crystal concentration and OD readings at 660 nm seen on the standard curve. Under similar experimental conditions, the results were comparable to the ones obtained by optical microscopy. The OD readings over 30 min revealed an instant decrease in the number of crystals, with maximum aggregation noted at 18 min. Addition of K-citrate at 1.25 mg/ml led to initial less crystal formation (OD = 0.236 nm vs. OD = 0.527 nm), with a maximum aggregation reached at 18 min. Overall, citrate addition decreased nucleation with a small change in the aggregation (OD = 0.316 vs. OD = 0.359). CONCLUSION: Spectrophotometric measurement of urinary turbidity is feasible and sensitive in assessing the potential clinical usefulness of different medications in inhibiting crystallization in urine.

Determination of struvite crystallization mechanisms in urine using turbidity measurement.

Sanitation improvement in developing countries could be achieved through wastewater treatment processes. Nowadays alternative concepts such as urine separate collection are being developed. These processes would be an efficient way to reduce pollution of wastewater while recovering nutrients, especially phosphorus, which are lost in current wastewater treatment methods. The precipitation of struvite (MgNH(4)PO(4)∙6H(2)O) from urine is an efficient process yielding more than 98% phosphorus recovery with very high reaction rates. The work presented here aims to determine the kinetics and mechanisms of struvite precipitation in order to supply data for the design of efficient urine treatment processes. A methodology coupling the resolution of the population balance equation to turbidity measurement was developed, and batch experiments with synthetic and real urine were performed. The main mechanisms of struvite crystallization were identified as crystal growth and nucleation. A satisfactory approximation of the volumetric crystal size distribution was obtained. The study has shown the low influence on the crystallization process of natural organic matter contained in real urine. It has also highlighted the impact of operational parameters. Mixing conditions can create segregation and attrition which influence the nucleation rate, resulting in a change in crystals number, size, and thus final crystal size distribution (CSD). Moreover urine storage conditions can impact urea hydrolysis and lead to spontaneous struvite precipitation in the stock solution also influencing the final CSD. A few limits of the applied methodology and of the proposed modelling, due to these phenomena and to the turbidity measurement, are also discussed.

X-ray microdiffraction and urine: a new analysis method of crystalluria.

The qualitative and quantitative analyses of crystalluria have clinical significance in the diagnosis and prognosis of urolithiasis. The aim of this paper is to provide a new accurate methodology to get qualitative and quantitative data on urine particulate in patients with renal stone disease.The procedure involves a urine collection, the separation of the solid residual by centrifugation, and its analysis by X-ray diffraction, utilizing a micro-diffractometer in order to analyze very low amounts of residual. The spectrum obtained was converted into 2 θ -I profiles and quantitatively refined by Rietveld method. The proposed methodology has the advantage to accurately quantify all crystalline phases and the amorphous component of the urine; anyway urine samples have to be centrifuged and analysed as soon as possible, because the quantitative results obtained by the X-ray microdiffraction showed that after some days and at room temperature urine increased significantly both amorphous and crystalline phases.

Risk factors and clinical presentation of cats with feline idiopathic cystitis.

Feline idiopathic cystitis (FIC) is the most common cause of feline lower urinary tract disease (FLUTD). This retrospective, case-controlled study evaluated possible risk factors associated with FIC and compared different clinical presentations in 64 cats with FIC. Several risk factors known to be involved in FLUTD were identified as playing a role in FIC. Of the stressful situations considered, most did not occur with increased frequency in cats with FIC compared to controls, except for a house move. The presence of pyuria, haematuria and an increased urine protein:creatinine ratio were significantly higher in obstructed males compared with non-obstructed males. An obstruction was significantly more likely in cats with struvite crystalluria compared with cats without struvite crystalluria. These findings suggest that urethral plugs might be an important cause or contributing factor of obstruction in FIC. Episodes of FIC seem to occur mainly in susceptible cats in combination with a deficient environment.

Effect of dietary water intake on urinary output, specific gravity and relative supersaturation for calcium oxalate and struvite in the cat.

It has been reported that daily fluid intake influences urinary dilution, and consequently the risk of urolithiasis in human subjects and dogs. The aim of the present study was to investigate the role of dietary moisture on urinary parameters in healthy adult cats by comparing nutritionally standardised diets, varying only in moisture content. A total of six cats were fed a complete dry food (6.3 % moisture) hydrated to 25.4, 53.2 and 73.3 % moisture for 3 weeks in a randomised block cross-over design. Urinary specific gravity (SG), urine volume, water drunk and total fluid intake were measured daily; relative supersaturation (RSS) for calcium oxalate (CaOx) and struvite was calculated using the SUPERSAT computer program. Cats fed the 73.3 % moisture diet produced urine with a significantly lower SG (P < 0.001) compared with diets containing 53.2 % moisture or lower. Mean RSS for CaOx was approaching the undersaturated zone (1.14 (sem 0.21); P = 0.001) for cats fed the diet with 73.3 % moisture and significantly lower than the 6.3 % moisture diet (CaOx RSS 2.29 (sem 0.21)). The effect of diet on struvite RSS was less clear, with no significant difference between treatment groups. Total fluid intake was significantly increased (P < 0.001) in the 73.3 % moisture diet (144.7 (SEM 5.2) ml, or 30 ml/kg body weight per d) compared with the 6.3 % (103.4 (SEM 5.3) ml), 25.4 % (98.6 (SEM 5.3) ml) and 53.3 % (104.7 (SEM 5.3) ml) moisture diets, despite voluntary water intake decreasing as dietary moisture intake increased. Cats fed the 73.3 % moisture diet had a higher total daily fluid intake resulting in a more dilute urine with a lower risk of CaOx when compared with the lower-moisture diets.

Lithogenic activity and clinical relevance of lipids extracted from urines and stones of nephrolithiasis patients.

We investigated contents and classes of urinary and stone matrix lipids, and evaluated their clinical relevance in nephrolithiasis patients. Lithogenic role of major lipid classes was explored. Urine (24 h) and stone samples were collected from 47 patients with nephrolithiasis. Control urines were obtained from 29 healthy subjects. Urinary 8-hydroxy-deoxyguanosine (8-OHdG), malondialdehyde (MDA), N-acetyl-ß-glucosaminidase (NAG) activity and total proteins were measured. Total lipids were extracted from centrifuged urines (10,000 rpm, 30 min) and stones by chloroform/methanol method. Major classes of lipids were identified using multi-one-dimensional thin-layer chromatography (MOD-TLC). Influence of each lipid class purified from stone matrices on stone formation was evaluated using crystallization and crystal aggregation assays. Urinary NAG activity and 8-OHdG were significantly elevated in nephrolithiasis patients. Total lipids in centrifuged urines of the patients were not significantly different from that of controls. In nephrolithiasis, urinary excretion of total lipids was linearly correlated to urinary MDA, 8-OHdG, NAG activity and total proteins. Lipid contents in stone matrices varied among stone types. Uric acid stone contained lower amount of total lipids than calcium oxalate and magnesium ammonium phosphate stones. MOD-TLC lipid chromatograms of healthy urines, nephrolithiasis urines and stone matrices were obviously different. Triacylglyceride was abundant in urines, but scarcely found in stone matrices. Stone matrices were rich in glycolipids and high-polar lipids (phospholipids/gangliosides). Partially purified glycolipids significantly induced crystal aggregation while cholesterol was a significant inducer of both crystal formation and agglomeration. In conclusion, total lipids in centrifuged urines did not differ between nephrolithiasis and healthy subjects. Our finding suggests that the significant sources of lipids in patients' urine may be large lipids-containing particles, which are removed in centrifuged urines. However, urinary lipid excretion in nephrolithiasis patients was associated with the extent of oxidative stress and renal tubular injury. Triacylglyceride was abundant in urines, but rarely incorporated into stones. Glycolipids were principal lipid constituents in stone matrices and functioned as crystal aggregator. Cholesterol purified from stone matrices bared crystal nucleating and aggregating activities.