Elemental Analysis of Urinary Calculi

Published: 2021-09-13 08:30:09
essay essay

Category: Learning, Zoology

Type of paper: Essay

This essay has been submitted by a student. This is not an example of the work written by our professional essay writers.

Hey! We can write a custom essay for you.

All possible types of assignments. Written by academics

The identification of the urinary calculi composition is essential as it provides information that could be useful for practitioners to find out the underlying cause of stone formation and to decide whether to treat the patients therapeutically or surgically. A study of the chemical composition of renal stones is important for understanding their etiology as well, permitting diagnosis and a proper management of the disease and the prevention of its recurrence.
Each canine stone sample was analysed for elemental content. Elemental analysis of stone sample can be perfomed by using X- ray diffraction (White et al., 1961; Leusmann et al., 1990), microradiographic techniques (Clark, 1976), transmission electron microscopy (TEM) (KENNOKI et al., 1977; Crawford, 1984), scanning electron microscopy (SEM) (Irving et al., 1986; Khan and Hackett, 1986; Delatte et al.,1987), energy dispersive X-ray (EDX) (Delatte et al., 1987; Reid et al., 1994; Marickar et al., 2009), Computed tomography (Hillman et al., 1984; Perlmutter et al., 2002), Infrared Spectrometry (Diaz-Espineira et al., 1997; Hawkins et al., 2009; Basiri et al.,2012), Fourier transform infrared (FTIR) spectroscopy (Marickar et al., 2009; Liu et al., 2011), Optical illuminated microscopy (Marickar et al., 2009), sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDSPAGE) (Liu et al., 2011), liquid chromatography/mass spectrometry/mass spectrometry (LC–MS/MS) (Liu et al., 2011). At present it is accepted that no single method provides total information on the structure and composition of the stone, and at least two different methods have to be combined for accurate study of calculi. Using combination of SEM and EDAX, the structure of stone crystal and amount of calcium, phosphorus, oxygen and other minerals present in the stone sample could be well understood (Marickar et al., 2009).
Animal screened
Twenty stone samples were obtained from veterinary clinical complex with physical information of clinical case (age, sex, breed and on outset of disease and reoccurrence) (Table 4.1). Radiographic and ultrasonographic examination and light microscopy ( for urine crystal visualization) were used for purpose of diagnosis of clinical case. Treatment protocol was surgical removal of urinary calculi in all twenty cases.
Diagnosis of cases under this study was done on the basis of history, physical examination, radiographic, ultrasonographic and laboratory finding (light microscopy of urine crystals).
A detail information was obtained from owner regarding patient with particular reference to any change in urinary habits i.e frequency of urination, 24 hour urinary output, any straining during urination, anuria, dysuria, haematuria, pain while urination, type of diet given and change in frequency of feed and water intake, Duration of illness whether accompanied by vomition.
Clinical sign exhibited by animal were change in urinary habit i.e frequency of urination, straining during urination, anuria, dysuria, haematuria, pain while urination and any sign of uraemia such as oliguria, constipation, vomiting, depression, loss of appetite, diarrhea and congestion of mucous membrane. In cases of partial or complete urinary obstruction of the urethra signs of bladder distention, abdominal pain, paradoxical incontinence, stranguria, and postrenal azotemia (anorexia, vomiting, depression) observed. Similar clinical finding observed by Langston et al ., (2008)
Anatomical site of stone lodgement was ascertained on preoperative diagnostic imaging with radiography and ultrasonography. Clinical signs exhibited by animals and distended abdomen with fluid thrills associated with radiography of abdomen and pelvic. In some cases ultrasonography were helped to reach diagnosis. Light microscopy was performed in few cases in which urine crystals were obtained.
Radiographic finding
Plain radiography of all Twenty clinical cases (male or female) were recorded in order to visualize caculi in urinary bladder or urethra. Radiography of caudoventral abdomen was carried out in dorsal and lateral recumbency which validate diagnostic method used by Shravanji, (2007). Plain lateral radiograph showing two large size urinary calculi lodged inside urinary bladder (Fig 4.1)
Ultrasonographic finding
Dogs suspected for urinary calculi were subjected to ultrasonographic examination. Ultrasonography carried out in dorsal or lateral recumbency with 3.5 MHz and 5 MHz microconvex transducer. Presence of cystic calculi was evident as hyperechoic focal echogenicity which formed a acoustic shadowing in the distal portion of bladder (Fig 4.2.2). Amount of acoustic shadowing varied with composition, density, compactness and number of calculi lodged in urinary bladder. Similar observation have been reported by Shravanji, (2007).
Light microscopy of urinary crystals
Urine samples from all twenty affected cases was obtained in clinical complex and light microscopic examination was done to visualize urinary crystal out of which nine patients showed crystals in urine (plate). Four types of crystals were obtained with different morphology i.e Calcium oxalate (colourless octahedrons), triple phosphate (rectangular coffin lid like), tyrosin (colourless brownish needles) and calcium carbonate (colourless spheres) (Fig 4.2.3). These findings also validate observation of Daudon et al ., (1993).
All dog patients were getting clinically required surgical treatment as used by Snyder et al ., (2005). No grouping in surgical treatment is followed. Choice of operative technique was governed by level of obstruction in lower urinary tract and physiological state of patient.
Patient was starved and water witheld for 12 hour before the operation. Operation site was shaved and scrubbed properly by using solution savlone and betadine.
Atropine sulphate (dose rate- 0.04 mg/kg body weight) was used intramuscularly as preanaesthetic medicant. Animal was anaesthesized by using Thiopentone sodium (5%) given I/V at the dose rate of 22 mg/kg body weight until desired depth of anaesthesia was obtained. Antibiotic was given before operation.
Retrograde urohydrorepulsion was performed as a part of preoperative treatment to push urethral calculi in the urinary bladder till cystotomy performed for removal of stone from urinary bladder. Before performing urohydrorepulsion bladder was evacuated by cystocyntesis in order to avoid bladder rupture. a flexible urinary catheter of appropriate size was inserted into lumen of urethra via external urethral orifice and advanced to site of obstruction as described by Osborne and Polzin, (1986).
A flexible urinary catheter (preferable a radio-marked infant-feeding tube) of appropriate size was inserted into lumen of urethra via external urethral orifice and advanced to site of obstruction. In small breed body weight upto 10kg, catheter size 6-7 was used, for medium sized breed body weight upto 15 kg, catheter size 7-8 was used and for large breeds body weight upto or more then 20 kg, catheter size 9-10 was used. Before insertion of catheter sterile lignocain gel was smeared on the surface of the catheter. Catheter is inserted by digital pressure. A 50 ml syringe loaded with normal saline was attached to hub of catheter and saline was flushed with optimum force into catheter. During entire procedure aseptic precaution was taken and catheter was kept intact till the day of cystotomy. Similar method was used by Osborne and Polzin, (1986).
Surgical methods used were cystotomy and/or urethrotomy, scrotal urethrostomy along with orchiectomy, tube cystostomy etc. according to postion of stone and condition of patient (Fig 4.3). cystotomy appeared to be a safe and effective surgical procedure for removal of lower urinary tract uroliths in most dogs, with only few, minor postoperative complications (Grant et al., 2010). Surgical approaches to cystotomy were – either by paramedian incision made lateral to prepuce (based on technique described by Weaver, 1970 ) or by ventral midline approach. Prescrotal Urethrotomy was performed based on the technique described by Stone (1990).
The intra-operative and post-operative anaesthetic and surgical events was recorded but for statistical analysis those events was considered as routine as this will remain beyond the scope of the objective of the study.
Post-operative Management includes administration of dextrose (5%) / Ringer’s lactate at 10-30 ml/kg body weight, intravenously, depending upon requirements for at least three days and postoperative antibiotics, for five days. The surgical wound was protected with antiseptic dressing until their complete healing in all cases. In all cystostomy cases, an indwelling Foley catheter of appropriate size was placed into the bladder for 7 days after operation, in order to facilitate the healing of the surgical wound of bladder and/or urethra. Skin sutures were removed on 10th postoperative day. Similar protocol was used by Shravanji, (2007)
Elemental analysis of urolith
Gross appearance
All urolith samples were cleaned manually with successive mild jets of NSS and distilled water. Gross appearance of all 20 calculi was observed for surface texture, luster, colour, shape, consistency of calculi (Fig 4.4.1)
All twenty urolith sample was of various diameters varying from 1 mm to 10 mm. Surface of stone sample was either smooth or rough. Colour of stones was white, yellow, brown, black. Shapes of urolith was different from round (Case No. 2830, 5215, 6443), elongated (Case No. 3127), botryoid (Case No. 1733, 2947, 2373), irregular (Case No. 3636, 5186, 13666, 6447, 5313, 5589, 6384) , triangular (Case No. 3114), branched (Case No. 1737) and papilliated. Similar findings were observed by Osborne et al., (1990) and Kale et al., (2011).
Scanning electron microscopic (SEM) analysis and Energy dispersive X- ray analysis (EDAX)
The SEM is especially useful for convenient inspection of urinary calculi. It produces three dimension- appearing images of micro structural features, and is a powerful tool for observing surface topography. Surface topography of the urolith samples was investigated with combination of scanning electron micrographs (SEM) and Energy dispersive X- ray analysis (EDAX) which require electron beam higher than 10 kv. Microscope used to obtain SEM images upto 30 kv acceleration beam with electron generated from tungsten source. Similar combination for stone analysis was used by Diaz-Espineira et al., (1995)
The SEM photographs were taken in different magnifications 80x to 500x. these photographs confirmed that the stone morphology are not a regular shape and size, it means that stone can have form different morphology and size in different samples. The microscopic analysis showed several concrements’ surfaces: almost smooth, hilly, with inclusions of crystals with keen edges, with roundish porous structure, united in conglomerates.
These images (Fig. shown different morphology of urinary stone i.e elongated, dumb bell, laminar bundle (Case No. 2373), pyramidal ((Case No. 2947), spherical (Case No. 1733, 5313), rhombohedral (Case No. 5589), irregular (Case No. 4769, 6443, 3127, 1463, 6384, 5125, 5215, 3259) crystal shapes. Similar morphological difference was observed by (Escolar and Diaz-Espineira et al., 1995; Bellanato, 1999; Samira et al., 2010; Kale et al., 2011 )..
EDAX is a technique of X- ray spectroscopy that is based on the collection and energy dispersion of characteristic X- rays. The X- ray spectrum consists of a series of peaks representative of the type and relative amount of each element in the sample. The number of counts in each peak is converted into elemental weight concentration either by comparison with standards or by simple calculation. With modern detectors and electronics most EDAX systems can detect X- rays from all the elements in the periodic table above beryllium (atomic number, Z = 4), if present in sufficient quantity. This property is helpful in analyzing mineral composition of urinary calculi. Similar technique for urinary stone analysis was used by Insight Knowledge, (2011).
These finding (Fig. shows the observed energy dispersive X-ray analysis (EDAX) results of the stone samples. A significant atomic and weight percentage of different element was observed in the measurement. EDAX result shows that these stone samples contain total 17 elements with different atomic and weight percentage can form different chemical compositions (minerals). Many chemicals compositions (minerals) contain H. However the lower atomic number (z ≤ 5) elements are not observed by EDAX technique, hydrogen falls in this category therefore hydrogen is not observed by EDAX.
Statistical analysis
The elemental analysis of urinary calculi of 20 dogs of different breeds, ages and sexes revealed that composition of calculi varied with each other with respect to level of presence or absence of different elements. However, (Chart. 4.5.1) clearly showed a very high percentage of “O” with an average of around 55% in each calculus while the other elements viz P, C, Ca and Mg were almost ranged between 8-15% in majority of calculi. Besides this, very low proportion of N, K, Cs, Na, Al, S, Cl and Fe was also recorded. The quantity of element like Ni, Ti and Zn was almost negligible. Interestingly, significant negative association was observed between Ca and P (r=-0.635**), Mg (r=-0.644**) and (r=0-.580**). Negative association was also recorded between P and C (r=-0.767**), Mg and C (r=-0.712**), O and K (r=-0.486*). On the other hand, significant positive correlations was observed between P and Mg (r=0.922**), P and K (r=0.657**), Mg and K (r=0.602**), Na and Si (r=0.494*), Na and S (r=0.605**), Na and Cl (r=0.448*), Si and S (r=0.882**) as well as Al and Fe (r=00.998**) (Table 4.5.1)
The correlation studies between different elements as urolith composition are not sufficient for clinical strategies for the prevention of urinary calculi in dogs. Hence, an attempt has been made for hierarchical clustering of different dog patients based on elemental groupings of urinary calculi. The idea of using mathematical methods in detection and definition of new disease entities is based on the following concepts suggested by Vogt and Nagel (1992).
A given data set for patients characterized by a disease entity considered until now as uniform is structured into different subgroups.
These subgroups correspond to different etiologies or forms of pathogenesis; or,
These subgroups differ from each other with respect to their therapeutic responsiveness or their prognosis.
Cluster analysis methods are appropriate to detect subgroups of entities. In cluster analysis a set of objects are grouped in such a way that objects in the same cluster are more similar to each other than to those in other clusters. The basic principle is the distance between the intra-clusters always lesser than inter-clusters. This can well be explained by dendogram (Chart. 4.5.2) having different clades where greater height of branch points indicating greater differences between the leaves. The horizontal axis of the dendrogram represents the distance or dissimilarity between clusters. The vertical axis represents the objects and clusters. So all 20 calculi can be grouped into clusters based on their compositional similarity (Chart. 4.5.3).
In the present findings the proximity matrix using squared Euclidean distance (Table 4.5.2) showed a vivid picture on closeness of different cases (dog patients) with each other where lesser the distance between the cases indicated closer the relation resulted in formation of different clusters. Again in agglomeration schedule (Table 4.5.3), the coefficient values between the clusters revealed that dissimilarity increased with the increase of the said values. Finally, in the dendogram different dog patients broadly classified into two groups where dog patients viz 3636, 3259, 2373, 5589, 5125, 5215, 4769, 6384, 1463,1366, 6443, 3114 and 1737 clustered in same groups while the patients of code 2947, 5186, 2830, 6447(2), 3127, 5313 and 1733 were in other groups. The formation of different clusters was based on elemental composition of urinary calculi of different dog patients. Based on similarity in composition of the urinary calculi the dog patients were further fell into different subgroups. These results indicated that formation of similar type of calculi in different dog patients might be due to their food habit, breeds, ages or sexes. These types of information are very useful in diagnosis for the treatment of such diseases.

Warning! This essay is not original. Get 100% unique essay within 45 seconds!


We can write your paper just for 11.99$

i want to copy...

This essay has been submitted by a student and contain not unique content

People also read