Bacterial isolates associated with urolithiasis in Thi-Qar province
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2021-08-14
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Abstract
Urolithiasis is a condition in which hard deposits of minerals and salts build inside the urinary system and can affect people of all ages all over the world. There are many factors related to this disorder including, age, gender, geographical area, environment and the life style. Stones in the urinary system are now considered one of the most serious issues that people face. There are few and limited number of studies related to urinary stones and bacterial isolates in Iraq , therefor this study aimed to isolate and identify bacterial isolates from these stones, as well as to assess antibiotic susceptibility, bacterial virulence factors, and urinary stone chemical analyses.
After surgery, stones were collected from 50 patients from different parts of the urinary system, including the kidneys, ureters, and urinary bladder. From October 2020 to April 2021, these samples were obtained from the urology units of Al-Hussein and Al-Habobi teaching hospitals as well as AL-Rahman private hospital in Al-Nasiriya city/Iraq. The age of patients ranges from 11 to 99 years old. The stones are crushed in a mortar and split into two parts: one for bacteriological study and the other for chemical study. The stone samples are inoculated on Nutrient broth and grown on blood and MacConky agar as part of the bacteriological investigation. This research includes isolating bacteria on traditional manual culture medium, which is done using old-school clinical microbiology lab culture procedures that are still considered the most essential method for identifying most bacteria.
The 76% of collected stones are from men while women account 24% of these stones. This result found that 67% of stone samples have bacterial isolates, whereas, the rest does not have bacterial isolates. The majority of total bacterial isolates which is related to urinary tract stones was Gram positive bacteria,
including Staphylococcus caprae (21.4), Micrococcus spp (21.4), Staphylococcus epidermidis (14.3), Staphylococcus xylosus (14.3), Staphylococcus aureus (7.14), Staphylococcus warneri (7.14), Staphylococcus capitis(7.14) and Staphylococcus auricularis (7.14). In addition, this study illustrates Gram negative bacteria from stones such as Ochrobactrum anthropi (10%) , E. coli 45%, Burkholderia cepecia (5%), Elizabethkingia meningosepticum (20%), Klebsiella pneumonia (5%), Pantoea septic (5%) , Chromobacterium violecium (5%) and Pseudomonas aeruginosa.
The antibiotics used are Amoxicillin/Clavulanic acid, Amoxicillin, ciprofloxacin,Vancomycin,Amikacin,Imipenem,Gentamicin, and Ceftriaxone. Detection and identification of bacterial isolates is based on each of biochemical tests, using API 20 test, measurement of sensitivity test of bacteria using susceptibility test to detect the therapy that target the specific pathogenic bacteria.
Gram-positive bacteria like Staphylococcus caprae, Staphylococcus epidermidis, and Staphylococcus xylosus were found to be sensitive to all antibiotics such as Imipenem, Amoxicillin, Amoxicillin/Clavulanic acid, Vancomycin, Gentamicin, and Ciprofloxacin 75 %, and resistant to Amikacin and Ceftriaxone 25 %. Staphylococcus warneri, Staphylococcus capitis, and Staphylococcus aureus were also shown to be completely resistant to all antibiotics. The Micrococcus is susceptible to 75 % of antibiotics, including imipenem, amoxicillin, amikacin, vancomycin, gentamicin, and Ciprofloxacin, and resistant to 25 % of antibiotics, including amoxicillin/clavulanic acid and Ceftriaxone.
Staphylococcus auricularis is responsive to Imipenem and Amoxicillin in 37.5 % of cases, but resistant to Amoxicillin/Clavulanic, Amikacin, Vancomycin, Gentamicin, and Ceftriaxone in 62.5 % of cases. Gram-negative bacteria, such as
Burkholderia cepecia, are completely resistant to all antibiotics, but Pantoea septic has a 100% susceptibility to all antibiotics. E.coli , Chromobacterium violecium and Pseudomonas.auruginosa was 25 % sensitive to Imipenem and Ciprofloxacin and 75 % resist to the other antibiotic . Ochrobacterium anthropi has a 25% resistance to Imipenem and Ceftriaxone and a 75% resistance to the other antibiotic; Chrysebacterium meningosepticum and Klebsiella pneumonia have a 12.5 % resistance to Imipenem and an 87.5 % resistance to the other antibiotic. These bacterial isolates are detected by using conventional PCR technique current rapid methods for detecting microbial pathogens in clinical specimens.
Virulence genes for Staphylococcus caprae are also considered by Atlc which the result was negative, while virulence genes for E. coli diagnosis are also considered by Fim , the result was positive . Finally, Pseudomonas aeruginosa virulence genes PAL1 and PAL2 were identified, as well as PS1 and PS2 for Pseudomonas fluorescens which the result was positive. The second part of the stones was investigated by using infrared spectroscopy examining the component of the stones. The findings revealed that uric acid prevalence increased by 76% and calcium oxalate prevalence increased by 24%, respectively.