Spot urine collection: A valid alternative to total urine collection for metabolomic studies in dairy cattle.

Urine is a highly suitable biological matrix for metabolomics studies. Total collection for 24-h periods is the gold standard as it ensures the presence of all metabolites excreted throughout the day. However, in animal studies, it presents limitations related to animal welfare and also due to alterations of the metabolome originating from the use of acid for preventing microbial growth or microbial contamination. In this study, we investigated whether spot urine collection is a practical alternative to total collection for metabolomic studies in lactating cows. For this purpose, we collected urine samples from 4 lactating Holstein cows fed 4 diets in a 4 × 4 Latin square design. Urine was collected for 24 h using a collecting device (i.e., total collection) or collected once per day 4 h after the morning feeding (i.e., spot urine collection). Dietary treatments differed by the amount of nitrogen content (high vs. low) and by the nature of the energy (starch vs. fiber). Urine metabolome was analyzed by 2 untargeted complementary methods, nuclear magnetic resonance and hydrophilic-interaction liquid chromatography (HILIC) coupled to a time-of-flight mass spectrometer, and by 1 targeted method, HILIC-tandem mass spectrometry. Although sampling technique had an effect on the abundance of metabolites detected, spot urine samples were equally capable of showing differences in urine metabolome than samples from total collection. When considering nitrogen levels in the diet, the robustness and precision for discriminating high- and low-nitrogen diets was equally achieved with both sampling techniques. A total of 22 discriminant metabolites associated with the N level of diets were identified from untargeted HILIC coupled to a time-of-flight mass spectrometer (n = 9) and nuclear magnetic resonance (n = 11), and 2 from targeted HILIC-tandem mass spectrometry. Alternatively, starch or fiber in the diet induced less changes in the metabolome that were not clearly discriminated independently of the sampling technique. We concluded that spot urine collection can successfully reveal differences in the urine metabolome elicited by dietary N levels and be used as a substitute of total urinary 24-h collection for metabolomic studies.

Effect of storage time on the urine protein: creatinine ratio in alkaline ovine urine.

The urine protein:creatinine (UPC) ratio is considered the reference method to assess proteinuria. Its diagnostic value in ovine medicine needs further elucidation. In population monitoring and/or for research purposes, it is convenient to collect many samples simultaneously and store them for later analysis. However, analyte stability data are required to ensure reliable results. We used 15 of 90 urine samples collected from sheep to assess the effect of storage time on the UPC ratio. After centrifugation, the supernatant of each sample was divided into 6 aliquots. Urine protein and creatinine concentrations were determined immediately in one aliquot using the pyrogallol red and a modified Jaffè method, respectively. The other aliquots were stored at -18°C. Based on the absence of active sediment, alkaline urine pH, and UPC ratio ≥0.2, we included 15 samples in our study. The UPC ratio was determined in the stored aliquots 2, 7, 14, 21, and 60 d after collection. The data were analyzed with univariate ANOVA. No significant difference was observed in the urinary concentrations of protein, creatinine, and the UPC ratio (0.8 ± 0.84 in conventional units and 0.09 ± 0.095 in SI units) among different times (p > 0.05). The UPC ratio remained stable for 2 mo in ovine urine samples stored at -18°C.

Estimating purine derivatives and nitrogen compound excretion using total urine collection or spot urine samples in grazing heifers.

The study aimed to evaluate the excretion of purine derivatives (PDs) and nitrogen compounds (NCs) and their ratios with creatinine in supplemented Zebu heifers kept on pastures by comparing total urine collection and spot sampling. Five Nelore heifers (400 ± 15 kg) were used in a 5 × 5 Latin square design. The treatments were the amount of concentrate (220 g of crude protein/kg dry matter) offered (0, 3, 6, 9 and 12 g/kg BW). In each period, the total urine collection was performed continuously for 3 days (subsampled at intervals of 4 h, 00:00-04:00 h, 04:00-08:00 h, 08:00-12:00 h, 12:00-16:00 h, 16:00-20:00 h and 20:00-24:00 h). The spot urine samplings were performed (in each period) for 24 h (0, 4, 8, 12, 16 and 20 h). Creatinine, total urinary nitrogen (UN), urea nitrogen (UreaN), allantoin and uric acid were analysed. Creatinine excretion was 23.01 ± 0.19 mg/kg BW and was not affected by collection day, treatment or their interactions (p > 0.05). Treatments affected (p < 0.05) PD excretions, however did not affect the ratio PD:creatinine (p > 0.05). Treatments and collection time affected (p < 0.05) NC excretion, whereas the UN:creatinine and UreaN:creatinine ratios were not affected (p > 0.05). Creatinine excretion and the PD:creatinine ratios in the urine samples estimated by the total or spot sampling were not different (p > 0.05). However, sampling method affected (p < 0.05) the UN:creatinine and UreaN:creatinine (p < 0.05) ratios. Creatinine can adequately estimate urinary excretion in grazing heifers, and a single spot urine sample at any time of the day can be used to estimate PD excretion in grazing heifers. But two spot urine samples are needed for proper NC excretion estimations in grazing heifers' urine.

Noninvasive sampling method for urinalysis and urine protein profile in captive giraffes.

Urinalysis could be helpful to investigate the health status of giraffes held in captivity using noninvasive methods to avoid animal handling or anesthesia. We collected 52 voided urine samples from 20 giraffes of different ages, sexes, and subspecies from the ground. To evaluate potential interference by soil contaminants, a pilot study was performed using 20 urine samples obtained from 10 cows. All bovine and 29 giraffe samples were subjected to routine urinalysis including urine specific gravity (USG). All samples were analyzed for urine total protein (uTP), urine creatinine (uCrea) concentration, and urine protein-to-urine creatinine ratio (UPC). Urinary proteins were separated by SDS-PAGE electrophoresis. No significant differences were determined between free-catch and urine sampled from the ground in cows. Giraffe urine was pale-yellow, with alkaline pH (>8.0) and a mean USG of 1.035 ± 0.013. The uTP, uCrea, and UPC expressed as median (range) were 0.20 (0.08-0.47) g/L, 2.36 (0.62-5.2) g/L, and 0.08 (0.05-0.15), respectively. SDS-PAGE allowed the separation of protein bands with different molecular masses, including putative uromodulin at 90 kD, putative albumin at 64 kD, and putative immunoglobulin heavy and light chains at 49 kD and 25 kD, respectively. Urine collection from the ground appears to be a reliable technique for urinalysis and urine electrophoresis in giraffes.

Urine cortisol-creatinine and protein-creatinine ratios in urine samples from healthy dogs collected at home and in hospital.

BACKGROUND: Recently, urine protein:creatinine ratios (UPC) were shown to be lower in urine samples from dogs collected at home (AH) as compared to those collected in hospital (IH). Stress-inducing procedures and travel to the hospital have been hypothesized to cause prerenal proteinuria. OBJECTIVES: Evaluate patient stress using urine cortisol:creatinine ratios (UCCr) and correlate UCCr to UPC in urine samples obtained AH and IH. ANIMALS: Thirty-six healthy, client-owned dogs. METHODS: Prospective, non-masked study. Two voided urine samples were obtained (AH and IH). Complete urinalysis as well as UPC and UCCr were performed. Clients graded their dogs' stress level AH, in transport, and IH. RESULTS: The UCCr was significantly higher in IH samples than in AH samples (P < .0001), but UPC was not significantly different between AH and IH urine samples (P = .14). In all samples and in both collection settings, UCCr was not significantly correlated with UPC. Travel time and time IH were not correlated with change in UCCr or UPC. In 8 dogs with borderline or overt proteinuria, no significant difference was found in UPC between settings, but UCCr was significantly higher in IH samples. CONCLUSIONS AND CLINICAL IMPORTANCE: The UPC was not higher when measured in urine samples collected IH compared to AH. Dogs had higher UCCr IH, but UCCr was not associated with UPC. Stress, as estimated by UCCr, did not affect proteinuria. Further evidence is needed to support the claim that stress may result in proteinuria in healthy dogs.

Does blood contamination of urine compromise interpretation of the urine protein to creatinine ratio in dogs?

AIMS: To determine the effect of contamination of urine with 0-5% blood, varying in haematocrit and protein concentrations, on the urine protein to creatinine ratio (UPC) in dogs, and to determine whether the colour of urine can be used to aid interpretation of UPC results. METHODS: Urine samples were collected by free catch from 18 dogs, all of which had UPC <0.2. Venous blood samples were also collected from each dog, and the blood from each dog was added to its own urine to produce serial concentrations of 0.125-5% blood. The colour of each urine sample was recorded by two observers scoring them as either yellow, peach, orange, orange/red or red. Protein and creatinine concentrations were determined, and dipstick analysis and sediment examination was carried out on each sample. Based on colour and dipstick analysis, samples were categorised as either having microscopic, macroscopic or gross haematuria. A linear mixed model was used to examine the effect of blood contamination on UPC. RESULTS: The uncontaminated urine of all 18 dogs had a UPC <0.2. Adding blood to the urine samples resulted in an increase in UPC at all contamination concentrations compared to the non-contaminated urine (p<0.001). None of the 54 samples with microscopic haematuria had UPC >0.5. For 108 samples with macroscopic haematuria the UPC was >0.5 in 21 samples (19.4 (95% CI=13.1-27.9)%), and for 54 samples with gross haematuria 39 (72 (CI=59.1-82.4)%) had a UPC >0.5. No samples had a UPC >2.0 unless the blood contamination was 5% and only 3/18 (17%) samples at this blood contamination concentration had a UPC >2.0. CONCLUSIONS AND CLINICAL RELEVANCE: This study showed that while blood contamination of ≥0.125% does increase the UPC, if the urine remains yellow (microscopic haematuria), then there is negligible chance that a UPC >0.5 will be solely due to the added blood. In that scenario, attributing the proteinuria present to the haematuria in the sample would be inappropriate. However blood contamination that results in discolouration of the urine sample from yellow (indicating macroscopic or gross haematuria) could increase the UPC above the abnormal range and would need to be considered as a differential for the proteinuria. Thus knowledge of urine colour, even if limited to simple colour scores (yellow, discoloured, red) could be utilised to aid interpretation of the UPC in samples with haematuria.

Evaluation of an ELISA for metanephrines in feline urine.

Catecholamines can be used to evaluate neuroendocrine tumors, stress, and potentially pain, but catecholamines degrade rapidly. Their metabolites normetanephrine (NME) and metanephrine (ME) have better stability in urine. In cats, urine sampling in a home environment would be beneficial to reduce effects of clinical stress and simplify sampling. We evaluated a human urine ELISA for analysis of NME and ME in feline urine, and investigated the effects of acidification, cat tray pellets, and storage time at room temperature up to 8.5 h. In 26 feline urine samples, mean NME concentration was 192 ± 80 ng/mL, mean intra- and inter-assay CV was 6.5% and 4.2%, respectively, and spike recovery was 98-101%, but dilutional recovery was unsatisfactory. For ME, mean intra- and inter-assay CV was 10.2% and 4.1%, respectively. Mean urine ME concentration was 32.1 ± 18.3 ng/mL, close to the kit's lowest standard, and spike recovery was 65-90%; the ELISA could not be validated for ME. The stability study, performed for NME on 12 urine samples, did not identify differences between acidified and non-acidified samples, cat tray pellets, or storage time, and no interaction effects. The ME ELISA was not suitable for feline urine; performance of the NME ELISA was acceptable, except for dilution recovery. For analysis of NME, feline urine can be sampled at home using cat tray pellets and stored at room temperature up to 8.5 h without acidification.

Idiopathic Dermal Necrosis in Black-tailed Prairie Dogs (Cynomys ludovicianus).

Because black-tailed prairie dogs (Cynomys ludovicianus) are used as a model for research on gallstones and bacterial infections, performing urinary evaluations can provide invaluable data. This case report involves 5 prairie dogs that developed moist necrotic skin lesions after urine collection by cystocentesis. The information presented here serves as a resource regarding a potential adverse event that may develop after cystocentesis in black-tailed prairie dogs.

Hydrophobic Sand Versus Metabolic Cages: A Comparison of Urine Collection Methods for Rats (Rattus norvegicus).

A common method for urine collection from rats requires the use of a metabolic cage, thus exposing animals to extended periods of isolation in an unfamiliar cage with a wire-mesh floor. A new method involving hydrophobic sand, a material more similar to bedding, has become available recently but has not been extensively compared with metabolic cages in regard to collection efficiency or stress. Using a within-subjects crossover design, we examined differences in stress markers, urinary markers, and urine volume of clinically healthy male Sprague-Dawley rats during 2-, 4-, and 6-h collection sessions in hydrophobic sand and metabolic cages. Stress response markers of weight loss, fecal pellet output, or corticosterone did not differ between hydrophobic sand and metabolic cages, and observed behavior suggested that sand may be less stressful than metabolic cages. All clinically relevant urinary markers examined were normal, with no differences between collection methods. Total urine volume collected was greater from the metabolic cage than sand in 3 of the 5 sessions, but the volume collected during the shortest session (2 h) did not differ between methods and accounted for 62% of the total volume collected during the longest session (6 h). Our results suggest that hydrophobic sand is a refinement of urine collection methods for rats that decreases isolation time, risk of injury, and stress and maintains the integrity of urine samples.

Effects of storage conditions on results for quantitative and qualitative evaluation of proteins in canine urine.

OBJECTIVE To investigate effects of storage conditions on the canine urine protein-to-creatinine ratio (UPC) and on SDS-agarose gel electrophoresis (AGE) of urinary proteins. SAMPLE Urine specimens from 20 proteinuric (UPC > 0.5) and 20 nonproteinuric (UPC ≤ 0.2) dogs. PROCEDURES UPC and SDS-AGE were performed on urine specimens stored at room temperature (20°C) and 4°C for up to 5 days and at -20° and -80°C for up to 360 days; some specimens were subjected to 3 freeze-thaw cycles. Results were compared with those obtained for fresh urine specimens. RESULTS UPC was not affected by storage at room temperature or by freezing. A decrease in UPC was observed for specimens from nonproteinuric dogs after 5 days at 4°C (10%) and from both groups after 90 days at -20° and -80°C (≤ 20% and ≤ 15%, respectively). The SDS-AGE profiles revealed no visual changes regardless of duration of storage for specimens stored at room temperature, 4°C, and -80°C, except for 1 profile after 360 days at -80°C. Repeated freeze-thaw cycles did not affect SDS-AGE profiles. Appearance or strengthening of high-molecular-weight bands that could alter interpretation was evident in SDS-AGE profiles after storage at -20°C for ≥ 15 days (31/40 dogs). CONCLUSIONS AND CLINICAL RELEVANCE Storage of urine at -20° or -80°C for up to 1 year influenced the UPC without affecting clinical interpretation. Storage of urine specimens at -20°C impaired visual analysis of SDS-AGE. When SDS-AGE cannot be performed on fresh or recently refrigerated urine specimens, storage at -80°C is recommended.

Avaliação dos perfis minerais séricos, urinários e sedimentares de ovinos recebendo dieta calculogênica / Evaluation of mineral profile in serum, urine and sediment of sheep receiving calculogenic diet

A urolitíase obstrutiva em pequenos ruminantes é uma doença metabólica de etiologia multifatorial com distribuição mundial. A elevação da concentração urinária de solutos, minerais ionizados (cristaloides) que formam cristais insolúveis é citada por alguns autores como o fator mais importante. Assim, o conhecimento do perfil mineral dos animais submetidos a dietas calculogênicas e a composição química dos urólitos tornam-se ferramentas eficazes na prevenção da doença. Neste estudo, foram utilizados 14 ovinos hígidos, machos (não castrados), da raça Santa Inês, com idade aproximada de 90 dias, distribuídos em dois grupos (G1 - sem vitamina C e G2 - com vitamina C) e alimentados com dieta calculogênica. A análise dos perfis minerais, séricos e urinários revelou completo desbalanceamento na relação entre concentrações de cálcio, fósforo e magnésio, havendo elevação expressiva do fósforo e do magnésio e diminuição substancial do cálcio. Com isso, a análise bioquímica dos urólitos demonstrou que o cálcio esteve presente em 50% das amostras analisadas.(AU)

Urolithiasis in small ruminants is a metabolic disease of multifactorial etiology with worldwide distribution. Increased urinary concentration of solutes, ionized minerals (crystalloid) that form insoluble crystals is cited by some authors as the most important factor. Thus, knowledge of mineral profile of the animals fed calculogenic diets and chemical composition of uroliths becomes an effective tool in preventing the disease. In this study, we used 14 healthy, male, non-neutered sheep, of the Santa Ines breed, aged approximately 90 days, divided into two groups (G1-without vitamin C and G2-with vitamin C) fed calculogenic diet. Analysis of mineral profiles in serum and urine revealed complete imbalance in the relationship between concentrations of calcium, phosphorus and magnesium, with significant increase of phosphorus and magnesium and substantial reduction of calcium. Thus, biochemical analysis of uroliths showed that calcium was present in 50% of samples.(AU)

Collection of untainted urinary specimens from the bladder of an anesthetized rabbit.

It is difficult to collect untainted urine specimens over short intervals of time during renal studies with rabbits. This is because both the ureters and the bladder of this species are relatively friable and minor manipulation can easily cause intraluminal bleeding. We have developed and refined an effective technique and protocol for placing an indwelling urinary bladder catheter into an anesthetized rabbit. The procedure is easy to perform and completely effective and reliable, allowing high-quality urinary specimens to be collected at intervals of 15-20 min over a period of 3-4 hours during a study of acute metabolic acidosis.

Effects of processing delay, temperature, and transport tube type on results of quantitative bacterial culture of canine urine.

OBJECTIVE: To determine the impact of processing delay, temperature, and transport tube type on results of quantitative bacterial culture (QBC) of canine urine. DESIGN: Diagnostic test evaluation. SAMPLE: 60 mL of pooled urine from 4 dogs, divided into six 10-mL aliquots. PROCEDURES: Urine aliquots were spiked with bacteria from 1 of 6 independent Escherichia coli cultures to achieve a target bacterial concentration of 10(5) CFUs/mL. One milliliter from each aliquot was transferred into 5 silicone-coated clot tubes (SCTs) and 5 urine transport tubes (UTTs). Samples were stored at 4°C (39°F) and 25°C (77°F) for 0, 8, and 24 hours, and then standard QBCs were performed. RESULTS: Median bacterial concentration for urine samples stored in a UTT for 24 hours at 4°C was lower than that for samples stored in an SCT under the same conditions. Conversely, a substantial decrease in median bacterial concentration was identified for samples stored for 24 hours in an SCT at 25°C, compared with the median concentration for samples stored in a UTT under the same conditions. Median bacterial concentration in samples stored in an SCT at 25°C for 24 hours (275 CFUs/mL) was less than the cutoff typically used to define clinically important bacteriuria by use of urine samples obtained via cystocentesis (ie, > 1,000 CFUs/mL). CONCLUSIONS AND CLINICAL RELEVANCE: Canine urine samples submitted for immediate QBC should be transported in plain sterile tubes such as SCTs. When prolonged (24-hour) storage at room temperature is anticipated, urine samples should be transported in UTTs.

Pilot Study: Colostomy and Urine Collection Protocol for Investigating Potential Inciting Causes of Hen Diuresis Syndrome.

Hen diuresis syndrome has emerged over the past 5 yr as a significant cause of mortality in the U.S. broiler breeder industry. The condition affects hens in production and is characterized by transient muscle weakness in the vent region, transient diuresis, and often urate deposits on the skin below the vent. Affected hens are often seen straining to lay an egg, which suggests oviduct contraction is also impaired. Related hen mortality, often reaching 1% or more a week, is believed to be primarily the result of male aggression of the vent region (Turner et al., "Investigating Causes of Excessive Urate Production in Broiler Breeder Hens Associated with Peritonitis and Cannibalism Mortality," Oral Presentation at The American Association of Avian Pathologists Annual Meeting, p. 139, 2010). The exact association between the cause of mortality and this syndrome is unknown, but it may be the consequence of transient partial to full oviduct prolapse, which predisposes or stimulates cannibalism and aggression. Based on unpublished work done prior to this study (Turner et al., ibid.), the evidence suggests the underlying problem is metabolic. We feel that urine collection and analysis is an essential component to understanding this condition. This study serves as a pilot study for future investigations that attempt to identify the nature and cause of the metabolic disturbance through paired urine and serum collection and analysis. For the purpose of this study, a small sample of 10 affected and 10 unaffected birds was used for sample collection. In order to collect pure urine, the birds were surgically colostomized. Colostomy did prove to be a useful means of collecting urine free of feces, and for the purposes of our study it yielded adequate urine samples for analysis. There were statistically relevant urine values observed. Affected birds had a higher presence of blood in the urine, a lower uric acid excretion rate (mg/hr), higher concentration (mEq/L) of urine Na+, and a lower concentration (mEq/L) of urine K+ than unaffected birds. This pilot study helps to address some of the pitfalls previously associated with colostomy and to determine when collection can begin postoperatively so that we can better understand when and how to begin our sampling in future trials to address the etiology of this condition.

Comparison between Urine Protein: Creatinine Ratios of Samples Obtained from Dogs in Home and Hospital Settings.

BACKGROUND: The urine protein:creatinine ratio (UPC) is used to quantify urine protein excretion and guide recommendations for monitoring and treatment of proteinuria. HYPOTHESIS/OBJECTIVES: Home urine samples will have lower UPCs than hospital samples. The objectives were to compare UPCs of samples collected in each setting and to determine whether environment of sample collection might affect staging, monitoring or treatment recommendations. ANIMALS: Twenty-four client-owned dogs. METHODS: Prospective, nonmasked study. Clients collected a urine sample from their dog at home and a second sample was collected at the hospital. Dogs receiving corticosteroids or angiotensin-converting enzyme inhibitors were excluded, as were those with urine samples of inadequate volume, no protein on dipstick analysis, or active urine sediment. Samples were refrigerated after collection, dipstick and sediment evaluations were completed and each sample was frozen at -80°C within 12 hours. UPCs were performed on frozen samples within 2 months. RESULTS: From 81 paired samples, 57 were excluded. Of the remaining 24, 12/24 (50%) had higher hospital sample UPCs, 9/24 (38%) had identical UPCs, and 3/24 (12%) had lower hospital UPCs. The UPCs of hospital samples were higher than home samples for the total population (P = .005) and the subset with UPC > 0.5 (P = .001). CONCLUSIONS: Setting and related circumstances of urine collection in dogs is associated with UPC differences; results are usually higher in hospital than in home samples. This difference has the potential to affect clinical interpretation.

Urine protein-to-creatinine concentration ratio in samples collected by means of cystocentesis versus manual compression in cats.

Objective-To compare urine protein-to-creatinine concentration (UPC) ratios in samples collected by means of cystocentesis versus manual compression in cats. Design-Evaluation study. Animals-43 client-owned cats requiring urinalysis. Procedures-In all cats, 5 mL of urine from the midstream phase of micturition was collected by means of manual compression and, subsequently, an additional 5 mL of urine was obtained by means of ultrasound-guided cystocentesis. A complete urinalysis was performed on all samples, and UPC ratios were determined. Results-Cats were classified on the basis of the International Renal Interest Society substaging system as being free from proteinuria (UPC ratio, < 0.2; n = 19) or as having borderline proteinuria (UPC ratio, 0.2 to 0.4; 7) or proteinuria (UPC ratio, > 0.4; 17). None of the cats had postrenal proteinuria. A significant linear correlation was identified between UPC ratios in urine samples obtained by means of manual compression and ratios in samples obtained by means of cystocentesis. For all cats, UPC ratios for samples obtained by the 2 collection methods resulted in classification in the same IRIS substage. Conclusions and Clinical Relevance-Results suggested that collection of a urine sample from the midstream phase of micturition by manual compression would be a reliable alternative to cystocentesis for the determination of UPC ratio in cats, provided that postrenal proteinuria was excluded by means of urine sediment analysis. Once postrenal proteinuria was ruled out, the method used to collect urine samples did not appear to influence the quantification of urine protein concentration.

Positive reinforcement methods to train chimpanzees to cooperate with urine collection.

Positive reinforcement training can be used in many ways to enhance the welfare of captive primates. Training for biologic sample collection is one application of positive reinforcement training. In this study, 35 adult female chimpanzees were trained to cooperate with the collection of urine samples needed to facilitate a research study. A median of 35 training sessions was required for the subjects to reach reliable performance (4 of 5 sequential attempts successful) of the urine collection behavior. Adult age had no effect on the speed of learning as indicated by a rank order correlation. Individual differences in the rate of learning were pronounced but did not vary with the age of the chimpanzees. Approximately 2 y after the initial training, and with continual sample collection taking place twice weekly, we assessed the reliability of their performance and found that the chimpanzees cooperated 100% of the time and that collection of a urine sample required about 5 min. Positive reinforcement training can markedly reduce staff time, particularly for studies such as this that require frequent biologic sample collection over long durations. Similar approaches could be used to train other laboratory primates to cooperate with urine collection procedures. Animal training programs that emphasize positive reinforcement training are an important refinement in the care of laboratory primates.

Non-invasive urine collection in the female southern hairy-nosed wombat (Lasiorhinus latifrons) with the aid of classical conditioning.

We propose that regular urine samples can be used to monitor and characterize the reproductive cycle of the wombat, but this approach has never before been attempted in a marsupial. We conducted a three stage conditioning process for non-invasive urine collection in captive female wombats, which included (1) initial habituation and observation of urination patterns; (2) classical association of a stimulus with urination and (3) urine collection with the classically-conditioned stimulus. Four of the five female wombats selected for this trial were successfully conditioned for urine collection. During stage 2, the animals urinated in response to tactile stimulation 96 times from 208 attempts (46%). In stage 3, urine was successfully collected 399 times from 485 attempts (82%), with the majority of samples being collected in the morning (280/388). Hand-raised females that were previously conditioned for toileting purposes as pouch young responded more rapidly to the stimulus than juvenile females with no prior conditioning. This study is the first description of a successful method of urine collection by classical conditioning in a marsupial.

A modified catheterization procedure to reduce bladder damage when collecting urine samples from Holstein cows.

This study proposed a modified procedure, using a small balloon catheter (SB catheter, 45 ml), for reducing bladder damage in cows. Holstein cows and the following catheters were prepared: smaller balloon catheter (XSB catheter; 30 ml), SB catheter and standard balloon catheter (NB catheter; 70 ml, as the commonly used, standard size). In experiment 1, each cow was catheterized. The occurrence of catheter-associated hematuria (greater than 50 RBC/HPF) was lower in the SB catheter group (0.0%, n=7) than in the NB catheter group (71.4%, n=7; P<0.05). In experiment 2, general veterinary parameters, urine pH, body temperature and blood values in cows were not affected before or after insertion of SB catheters (n=6). The incidence of urinary tract infection (UTI) was 3.0% per catheterized day (n=22). In experiment 3, feeding profiles, daily excretion of urinary nitrogen (P<0.05) and rate from nitrogen intake in urine (P<0.01), were higher with use of the SB catheter (n=13) than with the use of the vulva urine cup (n=18), indicating that using the SB catheter can provide accurate nutritional data. From this study, we concluded that when using an SB catheter, the following results occur; reduction in bladder damage without any veterinary risks and accuracy in regard to feeding parameters, suggesting this modified procedure using an SB catheter is a useful means of daily urine collection.