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Signifi cant Growth of E. coli in Mac Conkey Agar 

Signifi cant Growth of E. coli in Mac Conkey Agar 

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Abstract Introduction: Antibiotic resistance of urinary tract pathogens has increased globally. Updated knowledge of the antibiotic resistance patterns of uropathogens in the health institutes is important for the selection of an appropriate empirical antimicrobial therapy. The aim of this study was to evaluate the multi drug resistant urinary iso...

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... tract infection (UTI) is defi ned as bac teriuria along with urinary symptoms and is identifi ed by growth of a signifi cant number of organisms of single species in the urine. Urinary tract infections are common medical problems in children and are important cause of morbidity. UTI during childhood varies by age and gender. UTI commonly aff ects boys during the fi rst year of life 1 , but thereafter 3-5% of girls are aff ec ted1, increasing to 10% by the teenage years 2 . Although several diff erent microorganisms can cause UTI, including fungi and viruses, bacteria are the major causative organisms and responsible for more than 95% of UTI cases 3 . Treatment of urinary tract infections is compromised worldwide by the emergence of bacteria that are resistant to multiple antibiotics 4 . Overuse and use of incomplete course of antibiotics as well as empirical antimicrobial therapy has been the major contributing factor in the development of Multi Drug Resistant(MDR) bacteria 5 . Multidrug resistance is defi ned as resistance to two or more diff erent structural classes of antimicrobial agents 6 . There is growing concern regarding the resistance to uropathogens to antibiotics. The clinical impact of drug resistance may be great or insignifi cant, depending on the level of resistance, the site of infection, and the availability of eff ective, nontoxic therapeutic alternatives 7 . This prospective study was conducted to identify UTI due to multidrug resistant uropathogens among children and to evaluate empiric antibiotic therapy. This is the prospective study conducted at Kathmandu Model Hospital on Pediatric patients (1-15 years) attending for the treatment of suspected UTI case. Three hundred and seventy two urine samples were examined from clinically suspected urinary tract infection during the research period of six months (December 2011- May 2012) using culture and sensitivity tests. All the patient or parents of the patient were instructed carefully for collection of morning mid- stream urine specimens. They were given a sterile, dry and clean collection bottles for urine collection. All the urine specimens were processed within 30 minutes of collection. Culture of all urine specimens was done on 5% Blood Agar and Mac Conkey Agar plate utilizing semi- quantitative culture method (Fig 1). Shaking with hand to ensure a uniform suspension of bacteria vigorously mixed the urine specimens. Then, an inoculating loop of standard dimension was used to take up approximately fi xed and known volume (0.001ml) of mixed urine and placed on the center of the plate. The drop was spread in a line and then over the entire surface of the agar plate. After inoculation, the culture plates were incubated in an inverted position at 37 o C for 24 hours. 8 After 24 hours, the numbers of colonies were counted on each plate, which was multiplied by 1000 to calculate the number of organisms per ml in the specimen. Samples showing 10 5 or more organisms per ml of urine were taken as signifi cant. Colony counts less than this was considered as non-signifi cant 8 . Gram’s staining and various biochemical tests identifi ed pure culture of bacterial growth. Diff erent biochemical media used were Triple Sugar Iron Agar, Sulphite Indole motility Agar, Urease agar, Simmons’s Citrate Agar, Methyl Red / Voges Proskauer Test and Oxidation Fermentation medium. Catalase, Coagulase and Oxidase Tests were also performed. The antibiotics used as fi rst line drugs for Gram negative bacteria were Amoxycillin (10 mcg), Cefotaxime (30 mcg), Ciprofl oxacin (5 mcg), Cefi xime (5 mcg), Cotrimoxazole (1.25/23.75 mcg), Norfl oxacin (10 mcg), Nitrofurantoin (300 mcg) and Ofl oxacin (5 mcg) and those used for Gram positive bacteria were; Amoxycillin (10 mcg), Cefotaxime (30 mcg), Ciprofl oxacin (5 mcg), Norfl oxacin (10 mcg), Cotrimoxazole (1.25/23.75 mcg), Nitrofurantoin (300 mcg), Gentamicin (10 mcg) and Ceftriaxone (30 mcg). The antibiotics used as second line drugs were Ceftriaxone (30 mcg), Ceftazidime (30 mcg), Gentamicin (10 mcg), Amikacin (30 mcg), Chloramphenicol (30 mcg), Piperacillin/Tazobactam (100/10 mcg), Cefoperazone/ Sulbactam (50/50 mcg) and Meropenem (10 mcg). All the antibiotics were tested by Kirby Bauer’s Disc Diff usion Technique. The colonies picked up and suspended in Nutrient broth and adjusted turbidity to 0.5 Mc Farland standard. Within 15 minutes, a sterilized cotton swab was dipped into the adjusted suspension. Carpet culture was done by streaking the swab over the entire sterile Mueller Hinton agar plate (Fig 2). The antibiotic impregnated discs were placed on the surface of the agar plate and then incubated at 37 o C for 18 hours 9 . The diff erent inhibition zones were measured and interpreted the results on the basis of zone size compared with standard interpretive table given by manufacturer. The organisms which showed resistant to all fi rst line antibiotics except Nitrofurantoin were tested for second line drugs. Of the 372 urine samples processed, 60 (16.13%) samples showed signifi cant growth where as majority of samples i.e. 284 (76.34%) showed no growth and 28 (7.53%) showed growth of no signifi cance (Table 1). Out of 372 patients, 192 (51.61%) were male, while 180 (48.39%) were female. Of the total samples, 350 (94.09%) were from outdoor patients, 7 (1.88%) were from indoor patients and 15 samples (4.03%) were from emergency Department. The age group of 1-5 years had the maximum requests of 179 (48.12%) for urine culture, while the age group 11-15 years was second with 102 (27.42%) requests. Age group of 6-10 years was the least with 91 (24.46%) request. The age and gender wise distribution of children with UTI is shown in Table 2. UTI was commonly found in young female children of age 1-5 years. MDR isolates were common in young children (46.67%) of the 1-5 years age group. Children of age group 6-10 years showed three (5.0%) and age group 11-15 years showed only two (3.34%) MDR isolates. Of the total MDR isolates, 31.67 occurred in young female children of age 1-5 years. UTI was signifi cantly high in female children ( p =0.004) in comparison to male. There was no signifi cant diff erence seen in MDR infection between male and female ( p =0.8). However, the signifi cant diff erence was seen in MDR infection between age group 1 to 5 and above 5 year’s group ( p =0.0001) Gram negative bacteria were predominant; constituting 56 (93.33%) of the total 60 isolates and 33(55.0%) were MDR. Among Gram negatives, Escherichia coli were the most frequently isolated species with 49 (81.67%). Gram positive organisms constituted only 4 (6.67%) of total isolates, and 2 (3.33%) of them were MDR. Both MDR isolates were Enterococcus faecalis (Table 3). Among the common antibiotics used as fi rst line against gram negative isolates, nitrofurantoin showed a susceptibility of 55/56 (98.21%). Quinolones (Ciprofl oxacin, Norfl oxacin and Ofl oxacin) followed Nitrofurantoin with susceptibility of 38/56 (67.86%). Among the second line antibiotics used, Meropenem was found to be most eff ective drug with susceptibility of 16/17 (94.12%) followed by chloramphenicol and Amikacin with a susceptibility of 15/17 (88.24%). Most of the Gram negative isolates i.e. 45 (80.36%) were resistant to Amoxycillin (Table 4, 5). Among the gram positive isolates, Nitrofurantoin was the most eff ective drug with susceptibility of 4/4 (100%) which was followed by Cefotaxime and Norfl oxacin (Table 6). Out of 60 isolates, 24 (40.0%) isolates were resistant to >3 drugs where as only 3 isolates of Escherichia coli and one isolate of Salmonella Paratyphi were sensitive to all antibiotics used. Among 56 gram negative isolates, 31(51.67%) isolates were MDR whereas of the 4 gram positive isolates, 2 (50.0%) isolates were MDR (Table ...

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... [36] Shrestha et al. reported 66.67% of enterococci as MDR in urine from the neighbouring country Nepal. [37] In concurrence to most other studies from India, linezolid was found to be the most effective drug. [27,28,31] Less resistance was also showed towards nitrofurantoin and these two drugs can be the affectively used for treatment of UTI by MDR enterococci in this region. ...
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Published by CDC of ANJA College, Sivakasi. 102-114. ISBN: 978-93-83191-10-9.