Ultrasonographic localization and mineral profile of urinary calculi in young and adult bovine

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1 Indian J. Anim. Res., 51 (2) 2017 : Print ISSN: / Online ISSN: AGRICULTURAL RESEARCH COMMUNICATION CENTRE Ultrasonographic localization and mineral profile of urinary calculi in young and adult bovine Prachi E. Taksande*, N.S. Saini, J. Mohindroo, Tarunbir Singh and Ashwani Kumar Department of Veterinary Surgery and Radiology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana , Punjab, India. Received: Accepted: DOI: /ijar.8422 ABSTRACT The study was conducted on 13 male bovine animals suffering from obstructive urolithiasis to evaluate the diagnostic approach, surgical outcome and mineral composition of uroliths. The animals were divided in two groups, Group I (n=8) of young animals and Group II (n=5) of adult animals. B-mode ultrasonography was done for locating the uroliths in penile urethra. All the animals were subjected to urethrotomy followed by ultrasonography at the suspected site of urolith lodgement. Ammonium chloride as urinary acidifier was 50 mg/kg body weight orally for 60 days was found beneficial for preventing recurrence of uroliths. It was observed that urethra heals normally in three weeks if left unsutured and the majority of uroliths were present at tip of penis in Group I and in sigmoid flexure in Group II animals. Staphylococcus species were the most prevalent bacteria causing bovine urinary tract infections associated with urolithiasis in both groups. It was concluded that ultrasonography was efficient to localize uroliths in urethra in both groups of animals. On Fourier Transform Infrared Spectroscopy estimation, calcium carbonate, carbonate apatite in Group I whereas calcium carbonate in majority of Group II animals, were the most common types of minerals causing formation of uroliths. Key words: Bovine, Calcium carbonate, Ultrasonography, Urethrotomy, Urolithiasis. INTRODUCTION Urolithiasis is a common cause of lower urinary tract obstruction in male bovine (Gazi et al. 2015). It is one of the major disease conditions causing heavy economic loss to the farmers. The buffalo calves suffer more frequently from urolithiasis as compared to cow calves and adult bovine animals (Makhdoomi and Mohsin 2013). Uroliths may result from super-saturation, precipitation and crystallization of various minerals viz. magnesium ammonium phosphate hexahydrate, calcium carbonates etc (Griffith 1979). Cited literature reports various predisposing factors for urolithiasis such as nutrition, ph of urine (Samal et al. 2011), metabolic, hereditary, endocrine and urinary tract infection (Weaver and Pillinger 1975, Griffith 1979). Obstructive urolithiasis in bovine is an emergency surgical condition (Khan et al. 2011). Tentative diagnosis of obstructive urolithiasis can be made from history, clinical signs, per rectal examination, urinalysis, radiography, ultrasonography and haemato-biochemical examination (Parrah et al. 2011). Although urolith may be present anywhere in the urethra but distal sigmoid flexure has been reported as the most common site for urolith obstruction in bovine (Gasthuys et al. 1993). Early and accurate localization of urolith is important for deciding exact site of incision for urethrotomy. Radiography can determine the location and predict the type of urolith based on radio densities and other physical characteristics in small animals (Fromsa et al. 2011). However, it is not possible to *Corresponding author s gracy_8284@yahoo.com. radiograph the urethral region in adult bovine animals because of heavy musculature and anatomy of the sigmoid flexure (Gazi et al. 2014). Recently ultrasonography has been reported as a useful tool in cases of obstructive urolithiasis. It helps to locate the stone or concretions in urethra and to assess the status of urinary bladder i.e. distention or rupture (Magda 2006). Scanty literature is available on the ultrasonographic diagnosis of site of urolith obstruction and comparative mineral composition of the calculi in young and adult bovine. Knowledge of exact composition of calculi is helpful for deciding a strategy to prevent its recurrence and will prove helpful in future regarding selection of balanced ration for the live stock which is at risk of development of obstructive urolithiasis. Therefore, the present study was undertaken with the objectives to evaluate the diagnostic approach and surgical outcome in bovine urolithiasis and to determine the mineral composition of bovine uroliths in young and adult bovine patients suffering from obstructive urolithiasis. MATERIALS AND METHODS The present study was conducted on 13 male bovine animals suffering from obstructive urolithiasis. The animals were divided into two age groups; Group I (n=8) comprised of young calves with the mean age 10.68±2.99 months and Group II (n=5) comprised of adult animals with the mean age 61.20±10.80 months. History and clinical findings were recorded. For urinalysis and bacterial isolation, urine was

2 collected after catheterization during urethrotomy and abdominocentesis in case of bladder rupture. For haematological (Hb, TLC, TEC and DLC) and biochemical (BUN, Creatinine, Calcium, Phosphorus, Magnesium, Potassium, Sodium and Chloride) estimation, blood was collected from jugular vein. All the animals were subjected to B-mode ultrasonography using 5.0MHz (Curvi-linear) and 7.0 MHz (Linear) transducers for the examination of urinary bladder and urethra, respectively. For sedation Inj. Midazolam (Mezolam 0.02 mg/kg body weight (Neon Laboratories Ltd, Mumbai, India) was used. The site of urolith obstruction was identified on the basis of ultrasound examination of penile urethra. Local infiltration anesthesia at the site of suspected obstruction was achieved with 2% lignocaine. Urethrotomy was performed in lateral recumbancy and urinary catheter was placed. Urethral incision was left unsutured and surgical wound was closed using horizontal mattress suture pattern. The catheter was fixed at the prepuce using single interrupted suture. Post operative treatment included, analgesic and antinflammatory Inj. Meloxicam (Melonex 0.2 mg/kg body weight (Intas Pharmaceuticals Ltd, India) intramuscular for 3 days and antibiotic Inj. Ampicillin + Dicloxacillin (AC-Vet 10 mg/kg body weight (Intas Pharmaceuticals Ltd, India) intramuscular for 5 days and ammonium 50 mg/kg body weight as urine acidifier orally for 60 days. Urethrotomy followed by cystorrhaphy was done in one case, where urinary bladder was ruptured and Foley s catheter was placed inside the bladder after cystorrhaphy for 10 days. The skin sutures were removed on 12 th day and urinary catheter was removed after 3 weeks post operatively. The stones retrieved during urethrotomy were subjected to mineral analysis by Fourier Transform Infrared Spectroscopy (FTIR) technique to determine the exact mineral composition of the uroliths. RESULTS AND DISCUSSION The affected animals showed mild to moderate dehydration, anorexia and depression. History of urine obstruction varied from 2 to 10 days. Mild to moderate abdominal distention was recorded in majority of the animals (n=12). Severe abdominal distention was seen in only one case where the urinary bladder was found ruptured which could have resulted due to persistent urethral obstruction resulting in uroperitoneum (Gasthuys et al. 1993). In the present study, majority of obstructive urolithiasis was recorded in the winter season and early Table 1: Site of urethrotomy and location of stone in urethra Volume 51 Issue 2 (2017) 297 summer, between the months of December to May. This might be related to decrease in water intake during winter season and thereby causing super saturation of urinary solutes leading to lower urinary tract obstruction (Parrah et al. 2010). In Group I, all the animals were uncastrated buffalo calves with a mean age of 10.68±2.99 months. In Group II, three animals were uncastrated buffalo bulls and two were cattle bullocks all with a mean 61.20±10.80 months. Presence of a single or multiple calculi and concretions was confirmed on ultrasonographic examination in all animals (Fig 7-10). In Group I, uroliths were detected at the level of sigmoid flexure (n=2) or at tip of penis/ pre-scrotal urethra (n=6) (Table 1). Whereas, in Group II animals, uroliths were found lodged at sigmoid flexure region (n=3) or at the tip of penis/ pre-scrotal urethra (n=2) (Table 1, Fig 1-6). This was in accordance with findings of Saharan et al.2013 for animals of group I whereas it was in contrast to earlier reports where the most common site of urolith obstruction in adults (group II) has been reported to be sigmoid flexure (Gasthuys et al. 1993). In small ruminants or calves urethral process or tip of penis is reported as the common site of obstruction (Sharma et al. 2009). Postoperatively One animal of Group II removed the urinary catheter after three days of surgery and a swelling developed over ventral abdominal region due to subcutaneous accumulation of urine through the urethrotomy incision site which was managed by replacing the urinary catheter. On physical examination the colour of urine was light amber to yellow with an alkaline ph within range of in both groups of animals. The microscopic examination of urine revealed few deposits in two animals of Group I and in one animal of Group II. Presence of pus cells was recorded in seven animals of Group I and in three animals of Group II. Urinary casts were seen in two animals of Group I and in one animal of Group II. The urine sediment did not reveal presence of crystals in any of the animals of either group. The urine culture revealed presence of Staphylococcus species in 5 animals of Group I and in 3 animals of Group II. According to Kushwaha et al. 2012, Staphylococcus species was commonly isolated bacteria from the urine samples of buffalo calves. Staphylococcus species is the most prevalent bacteria in small animals that have been reported to cause urinary tract infections and is Group Urethrotomy Location of stone No. of animals Group I Pre-scrotal Tip of penis/ Pre-scrotal urethra 6 Post-scrotal Level of sigmoid flexure 2 Group II Pre-scrotal Tip of penis/ Pre-scrotal urethra 2 Post-scrotal Sigmoid flexure 3

3 298 INDIAN JOURNAL OF ANIMAL RESEARCH Fig 1: Photograph showing pre-scrotal urethrotomy to remove single urolith at tip of penis Fig 2: Photograph showing post-scrotal urethrotomy to remove urolith at the level of sigmoid flexure Fg 3: Photograph showing post-scrotal urethrotomy to remove urolith removal ½ inch above retractor penile muscle Fig 4: Photograph showing concretions at tip of penis Fig 5: Photograph showing urolith removal at about 3cm from the tip of penis Fig 6: Photograph showing site of ruptured urinary bladder

4 Volume 51 Issue 2 (2017) 299 Fig 7: Ultrasonogram showing two stones at tip of penis (Hyper echoic areas with acoustic shadow) Fig 8: Ultrasonogram showing distended urethra caudal to obstruction site Fig 9: Ultrasonogram showing concretions at tip of penis and distended urethra caudal to site of obstruction an important predisposing factor for initiation, growth and recurrence of struvite crystals (Osborne et al. 1981, Gatoria et al. 2006). However, struvite was only reported in one animal of Group I in present study. This indicated that in large animals the Staphylococcus species may cause urinary tract infection predisposing other type of calculi like calcium carbonate and carbonate apatite. Primarily treatment of obstructive urolithiasis is surgical management which consists of urethrotomy, cystotomy or urethrostomy. Surgical treatment is required in most of cases during the treatment process of obstructive urolithiasis (Jennifer et al. 2008, Kalim et al. 2011). In present study also, urethrotomy was done in all animals at different sites depending upon the location of stone as determined by ultrasonography (Fig 7-10). Unsutured Fig 10: Ultrasonogram showing single urolith 3cm caudal to tip of penis urethra with presence of catheter for 3 weeks prevented formation of urethral stricture and facilitated the healing of urethral wound. Similar findings were also reported by Anjaria (1969). Based on location of urolith, pre-scrotal urethrotomy was done in six animals of Group I and two animals of Group II. Post-scrotalurethrotomy was done in two animals of Group I and in 3 animals of Group II. The present study indicated that there are more chances of urolith obstruction at tip of penisor pre-scrotal urethra in young animals (Group I) and according to previous reports though sigmoid flexure is the common site (Gasthuys et al. 1993) but present study suggests that uroliths can also lodged at tip of penis in adult animals (Table 1). Ammonium chloride given to all animals as urinary acidifier was found beneficial for preventing recurrence of uroliths (Singh et al. 2010) in both groups.

5 300 INDIAN JOURNAL OF ANIMAL RESEARCH The mean hemoglobin, TEC and TLC values of both groups were within normal range (Table 2). An increase in neutrophil count was recorded in both groups. This relative increase in neutrophil may be due to the prolonged infection in the urinary tract. Sharma et al also reported an increased neutrophil count in urolith cases of bovines. The mean BUN and creatinine values of Group I and II were above normal range (Table 2) which may be due to prolonged accumulation of urine and consequent uraemia. The BUN (179.8) was markedly elevated in one animal of Group I, which had bladder rupture along with high Creatinine (6.12) value which may be due to uroperitoneum. One animal of Group II with intact bladder had a high creatinine value (18.2), recovered after surgery. Increased BUN and creatinine in cases of urinary obstruction in bovine was also reported by Sharma et al The magnesium, calcium and sodium levels of the animals of both groups of animals were within normal range (Table 2). The mean phosphorus value of Group I animals was above normal reference range but in animals of Group II, the phosphorus was within the normal reference range (Table 2). The phosphorus value was markedly elevated in two animals of Group I. The hyperphosphatemia in these animals may be due to decreased glomerular filtration rate and imbalance of Ca:P ratio. There is an inverse relation between calcium and phosphorus. The phosphorus level in blood increases if there is a decrease in calcium level in blood (Sharma et al. 2005). Bovine urolithiasis mostly occurred if the normal calcium phosphorus ratio (2:1) is imbalanced which was also observed in present study in which the Ca:P ratio was found to be 1.41:1 in both groups of animals. Vinodhkumar et al also reported that animals suffering from urolithiasis had slightly higher phosphorus and lowered calcium in serum. The potassium levels in all the animals were towards the high range in both groups. This may be attributed to the urine retention as potassium is mainly excreted via kidneys in bovines (Sharma et al. 2005). The chloride level was within normal range in both groups. Table 2: Hematology and bio-chemical profile in young (Group I) and adult (Group II) animals Uroliths are formed due to imbalanced intake of minerals particularly high level of phosphorus and magnesium with low level of calcium and potassium in feed and concentrate rich diet (Parrah et al. 2011). Deprived water intake also plays a major role in bovine urolithiasis. Grossly the uroliths retrieved during urethrotomy were single or multiple stones in 5 cases and sandy concretions in 3 cases in Group I. Majority of the uroliths were white in colour with rough surface and irregular margins with an approximate size of 0.6 cm to 1cm in length and 0.3 cm to 0.6 cm in diameter (Fig 11). In Group II animals, single white to dull brown in colour urolith with rough surface and irregular margins with an approximate size of 0.5 cm to 1.1 cm in lengthand 0.3 cm to 0.5 cm in diameter (Fig 11) were retrieved. The mineral analysis of the uroliths done by FTIR technique retrieved during urethrotomy, revealed presence ofstruvite in one animal of Group I having high serum level of phosphorus. In four animals of each group, calcium carbonate was the found to be the main mineral component of uroliths (Table 3). Carbonate apatite which contains considerable amount of calcium phosphate (Dahllite) was present in three animals of Group I and in one animal of Group II. The most common types of minerals causing urolithiasis found in ruminantsare struvites, calcium oxalate, calcium phosphate, calcium carbonate and silica (Parrah et al. 2010, Samal et al. 2011, Makhdoomi and Mohsin 2013) but in present study calcium carbonate and carbonate apatite were found to be the major components causing bovine urolithiasis (Table 3). Table 3: Mineral analysis of stones in obstructive urolith cases Mineral analysis Group I Group II Struvite + Carbonate apatite Calcium carbonate Carbonate apatite Total Parameter Group I Group II Normal range Hb (g %) 11.42± ± TLC (/cu mm) ± ± ,000 13,000 TEC ( 10 6 /cu mm) 8.13± ± DLC (/cu mm) N 60.25± ± L 38.75± ± M 2.50± ± BUN (mg/dl) 70.35± ± Creatinine (mg/dl) 4.70± ± Magnesium (mg/dl) 2.16± ± Calcium (mg/dl) 8.66± ± Phosphorus(mg/dl) 8.02± ± Sodium (meq/l) ± ± Potassium (meq/l) 6.45± ± Chloride (meq/l) ± ±

6 Volume 51 Issue 2 (2017) 301 Fig 11: Showing gross physical characteristics of uroliths in bovine It was concluded that ultrasonography is efficient to diagnose the obstructive urolithiasis with rupture of urinary bladder in cattle and buffaloes. Ammonium chloride as a urinary acidifier was found beneficial for preventing recurrence of uroliths in young and adult animals. Uroliths are mostly present at tip of penis in young and in the region of sigmoid flexure in adult bovine. Male uncastrated buffalo calves are more prone to obstructive urolithiasis in winter. Urethra heals normally if left unsutured in three weeks interval without any postoperative complications.. Animals suffering from obstructive urolithiasis were having disturbed Ca:P ratio. Staphylococcus species are the most prevalent bacteria causing bovine urinary tract infections associated with urolithiasis Calcium carbonate and carbonate apatite are the most common type of minerals causing formation of uroliths in young and calcium carbonate in majority of adult bovine animals. REFERENCES Anjaria, J. V. (1969). Observations on bovine urethral calculosis. IVJ 46: Fromsa, A., Saini, N. S and Rai, T. S. (2011). Diagnosis, prediction and mineral analysis of uroliths in canines. GV 7: Gasthuys, F., Steenhaut, M., Moor, A. D and Sercu, K. (1993). Surgical treatment of urethral obstruction due to urolithiasis in male cattle: a review of 85 cases. Vet Rec 133: Gatoria, I. S., Saini, N. S., Rai, T. S and Dwiwedi, P. N. (2006). Comparison of three techniques for the diagnosis of urinary tract infections in dogs with urolithiasis. J Sm Anim Prac 47: Gazi, M A., Khan, M A., Makhdoomi, D. M., Siraj, G., Dar, S. H and Mir, A Q Possible role of calcium, phosphorus and magnesium shift in blood, urine and calculi in calves affected by urolithiasis. African Journal of Agricultural Research 10: Gazi, M A., Makhdoomi, D. M., Parrah, J. D., Ganai, A. M., Sheikh, G. N and Mir, S, A Recent advances in surgical management of urolithiasis in sheep and goat. African Journal of Agricultural Research 9: Griffith, D. P. (1979). Urease stones. Urol Res 7: Jennifer, M. E., Meredyth, L. J and Matt, D. M. (2008). Surgery of obstructive urolithiasis in ruminants. Vet Clin North Am: Food Anim Prac 24:

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