Maria Sindhura John, J Biosci Tech,Vol 6(1),2015,620-626 ISSN: 0976-0172 Journal of Bioscience And Technology www.jbstonline.com Prevalence and antibiotic susceptibility pattern of bacterial pathogens causing urinary tract infections in humans at a tertiary care hospital in AP N.Premanatham1, Maria Sindhura John*2, P.Muni Lakshmi3 and P.Sreenivasulu Reddy4 1. Associate Professor, 2. Tutor, 3. Assistant Professor 4. H.O.D Dept. of Microbiology Narayana Medical College, Nellore, A.P. *Corresponding author, E.Mail: sindhura .john @gmail.com. ABSTRACT: BACKGROUND: Urinary tract infections are most common infection caused by bacterial pathogens seen mostly in developing countries. It is one of the common clinical conditions in the patients presenting to the hospitals. A causative agent varies from place to place. OBJECTIVE: Studies are done to know the most common pathogen of urinary tract infections and its antibiotic susceptibility patterns, which is helpful to the clinicians. MATERIAL AND METHODS: A total of 200 mid-stream urine samples collected from the suspected UTI patients were tested microbiologically on CLED agar and antimicrobial susceptibility test were performed for the isolated pathogens using Kirby-Bauer disk diffusion method. RESULTS: The rate of culture positivity in females was 91.6% and in males was 40.3%. E-coli was the most frequently isolated urinary pathogen (42.8%), followed by Klebsiella(16.8%) and Acinetobacter (12.9%).E-coli was highly sensitive to Nitrofurantoin(72.7%) and followed by Gentamycin, Imipenam, Nalidixic acid. and it was highly resistant to Ampicillin(3%). Klebsiella was highly sensitive to Impinem (92.3%) and it was highly resistant to Ampicillin. CONCLUSION: Higher prevalence of UTI was seen in females. Gram negative organisms were the most commonly isolated in UTI than gram positive organisms. Among which E.coli is most commonly isolated orgaism showing high sensitive to nitrofurantoin followed by Klebsiella and Acinitobacter. 1. INTODUCTION A urinary tract infection, or UTI, is an infection of the urinary tract. The infection can occur at different points in the urinary tract. An infection in the bladder is also called cystitis or a bladder infection. An infection of one or both kidneys is called pyelonephritis or a kidney infection. The tubes that take urine from each kidney to the bladder are only rarely the site of infection. An infection of the tube that empties urine from the bladder to the outside is called urethritis. Urinary tract infections are the most common infections in clinical practice. (1) Urinary tract infection (UTI) is one of the most important causes of morbidity in the general population, and is the second most common cause of hospital visits. (2) KEYWORDS: Urinary tract infections, Bacterial pathogens, Midstream, Nitrofurantoin, CLED Uncomplicated UTIs typically occur in the healthy adult non-pregnant woman, while complicated UTIs (cUTIs) may occur in all sexes and age groups and are frequently associated with either structural or functional urinary tract abnormalities. Examples include foreign bodies such as calculi (stones), indwelling catheters or other drainage devices, obstruction, immunosuppression, renal failure, renal transplantation and pregnancy.(3) UTIs is described as a bacteriuria with urinary symptoms (4). The lower UTI infection is characterized by symptoms such as dysuria, frequency, urgency, and suprapubic tenderness.(5). UTI is more common in females than in males as female urethra structurally found less effective for preventing the bacterial entry (6). It may be due to the proximity of the genital 620 Maria Sindhura John, J Biosci Tech,Vol 6(1),2015,620-626 tract and urethra (7) and adherence of urothelial mucosa to the mucopolysaccharide lining (8). The other main factors which make females more prone to UTI are pregnancy and sexual activity (9). In pregnancy, the physiological increase in plasma volume and decrease in urine concentration develop glycosuria in up to 70% women which ultimately leads to bacterial growth in urine (10). Also in the nonpregnant state the uterus is situated over the bladder whereas in the pregnant state the enlarged uterus affects the urinary tract (11). Sexual activity in females also increases the risk of urethra contamination as the bacteria could be pushed into the urethra during sexual intercourse as well as bacteria being massaged up the urethra into the bladder during child birth (12, 13). Using a diaphragm also causes UTI as it pushes against the urethra and makes the urethra unable to empty the bladder completely and the small concentration of urine left in the bladder leads to the growth of bacteria which ultimately causes UTI (14). UTI is frequently encountered in patients with diabetes and in those with structural and neurological abnormalities, which interfere with urinary flow. The prevalence in both out and hospital patients with UTI is increasing and can vary according to geographical and regional location. (15) The aim of the study is to determine the prevalence of UTI in male and female patients as well as the effect of gender and age on its prevalence. The UTI-causing microorganisms, their distribution among different ages and genders, also be determined. The quantitative criterion appropriate for the microbiological identification of significant bacteriuria is generally considered to be at ISSN: 0976-0172 Journal of Bioscience And Technology www.jbstonline.com least 108 cfu/L. In some specific groups it is less: for men ≥106 cfu/L; and for women with symptoms of UTI it is ≥105 cfu/L. (16) The most common pathogenic organisms of UTI are Escherichia coli, Staphylococcus saprophyticus and less common organisms are Proteus sp., Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterococci and Candida albicans.(17).Although a broad range of pathogens can cause UTI, Escherichia coli remains the most common; however, even this organism is becoming resistant to the agents that are normally prescribed. Drugs commonly recommended for simple UTIs are Sulfamethoxazole-trimethoprim (Bactrim, Septra, others) Amoxicillin (Amoxil, Augmentin, others) Nitrofurantoin (Furadantin, Macrodantin, others) Ampicillin, Ciprofloxacin (Cipro), Levofloxacin (Levaquin) Usually, symptoms clear up within a few days of treatment. But you may need to continue antibiotics for a week or more. Take the entire course of antibiotics prescribed by your doctor to ensure that the infection is completely gone. 2. MATERIAL AND METHODS This prospective study was conducted over a period of six months. During this period a total of 200 urine samples were collected in sterile containers from suspected urinary tract infected cases from different clinical departments in Narayana Superspeciality Hospital, Nellore, A.P. Work is carried out in Department of Microbiology, Narayana Medical College, Nellore, AP. Mid-stream urine samples from the suspected UTI patients referred by physicians are taken. The urine samples were collected into labelled 20ml calibrated sterile bottles distributed to 621 Maria Sindhura John, J Biosci Tech,Vol 6(1),2015,620-626 the patients by the attending physicians suspected to have UTIs. In each container, boric acid (0.2 mg) was added to prevent the growth of bacteria in the urine. All patients were instructed on how to collect the urine samples aseptically and taken to the laboratory immediately for culture. The collected urine samples were inoculated on CLED (cysteine lactose electrolyte deficient agar) agar plates and cultured media were incubated aerobically at 37°C. The urine culture plates were examined for pure growth determined by morphologically same type of colonies and colony counts for determination of significant and insignificant growth. A growth of ≥105 colony forming units/ml was considered as significant bacteriuria. Identification of bacterial pathogens was made on the basis of Gram reactions, morphology, biochemical characteristics and cultural characteristics. Gram staining was performed to differentiate the Gram positive and Gram negative organisms. Antimicrobial sensitivity of the confirmed micro-organisms was done by disc diffusion method on Muler Hinton agar. Antibiotic susceptibility tests and interpretations were carried out for bacterial isolates by the KirbyBauer technique. The antibiotics tested were Nitrofurantoin, Amikacin, Cotrimoxazole, Gentamycin, Ciprofloxacin, Cefoxitim, Nalidixic acid, Norfloxacin, Ampicillin and Impinem for gram negative organisms For gram positive organisms drugs tested were penicillin, amoxicillin, Doxycyclin, vancomycin, oxacillin, linezolid, cefoxitin. 3. RESULTS The study was done from June 2014 to December 2014 at the Department of Microbiology, Narayana Medical College, Nellore, AP, and India. The name, age, sex and address of the patients was also recorded. ISSN: 0976-0172 Journal of Bioscience And Technology www.jbstonline.com Pathogenic bacteria were isolated in 200 samples with a prevalence rate of 77%.The prevalence in females was 91.6% and the prevalence rate in males was 40.3%, theprevalence pattern in males and females were shown.(Table-1) A total of 200 samples were collected in the study period.among which 143 (91.6%) were from females and 57(40.3%) samples were from males.131 samples are positive among females and 23 samples are positive among males. Highest no.of samples are collected from females. (Table-1) Based on age distribution maximum no.of samples collected from 21-40 yrs age were 101 samples (50.5%). UTI was most commonly seen in the age group. Minimum no.of samples collected from age group >80 were 9 samples(4.5%).below age group 20 yrs the samples collected were 47 (23.5%).(Table-2) Among the pathogenic bacteria isolated, Ecoli was the most frequently isolated urinary pathogen (42.8%), followed by Klebsiella (16.8%), Acinetobacter (12.9%) and candida (11%). Pseudomonas and Proteus was isolated as1.9%, 0.6% respectively(Table- 3) Age and gender wise distribution of pathogens causing urinary tract infection is clearly shown. Among the age group 21-40 yrs 87 (86.1%) samples were shown positive. Among them highest positivity is shown by females, 80samples than males 7samples. Least no.of samples were collected from >80 yrs,3 samples. Among them females showed positivity of 2 samples, while men showed 1 sample. (Table-4) E.coli was highly sensitive to Nitrofurantoin. and it was highly resistant to Ampicillin. 622 ISSN: 0976-0172 Journal of Bioscience And Technology www.jbstonline.com Maria Sindhura John, J Biosci Tech,Vol 6(1),2015,620-626 TABLES 1. SEX WISE DISTRIBUTION S.NO SEX 1 2 MALE FEMALE TOTAL NO.OF SAMPLES PROCESSED 57 143 200 NO.OF POSITIVE SAMPLES 23 131 154 PERCENTAGE % 40.3% 91.6% 77% 2. AGE DISTRIBUTION AGE TOTAL SAMPLES COLLECTED <20 21-40 41-60 61-80 >80 TOTAL 47 101 27 16 9 200 PERCENTAGE % 23.5% 50.5% 13.5% 8% 4.5% 100% 3. URINARY PATHOGENS ISOLATED PATHOGENS ISOLATED E.coli Klebsiella Acinitobacter Candida Citrobacter CONS Staphylococcus Pseudomonas Proteus TOTAL NO. OF PATHOGENS 66 26 20 17 10 6 5 3 1 PERCENTAGE% 42.8% 16.8% 12.9% 11% 6.4% 3.8% 3.2% 1.9% 0.6% 4. AGE AND GENDER WISE DISTRIBUTION OF URINARY TRACT INFECTION AGE <20 21-40 41-60 61-80 >80 TOTAL NO.OF POSITIVE SAMPLES MALE FEMALE 9 28 7 80 5 13 1 8 1 2 23 131 TOTAL PERCENTAGE% 37/47 87/101 18/27 9/16 3/9 154/200 78.7% 86.1% 66.6% 56.2% 33.3% 77% 623 ISSN: 0976-0172 Journal of Bioscience And Technology www.jbstonline.com 5. ANTIBIOTIC SUSCEPTIBILITY PATTERN OF GRAM NEGATIVE ISOLATES Maria Sindhura John, J Biosci Tech,Vol 6(1),2015,620-626 DRUGS Nitrofurantoin Amikacin Cotrimoxazole Gentamycin Ciprofloxacin Ceftriaxime Nalidixic acid Ampicillin Imipenam Amoxyclav E.COLI 48/66 (72.7%) 20/66 (30.3%) 10/66 (15.1%) 32/66 (48.4%) 15/66 (22.7%) 12/66 (18.1%) 40/66 (60.6%) 2/66 (3%) 42/66 (63.6%) 6/66 (9%) KLEBSIELLA 23/26 (88.4%) 16/26 (61.5%) 8/26 (30.7%) 6/26 (23%) 3/26 (11.5%) 2/26 (7.6%) 20/26 (76.9%) 0/26 (Nil) 24/26 (92.3%) 6/26 (23%) ACINITOBACTER 8/20 (40%) 12/20 (60%) 6/20 (30%) 10/20 (50%) 4/20 (20%) 3/20 (15%) 13/20 (65%) 0/20 (Nil) 14/20 (70%) 8/20 (40%) 6. ANTIBIOTIC SUSCEPTIBILITY PATTERN OF GRAM POSITIVE ISOLATES DRUGS Penicillin Amoxicillin Doxycyclin Cefoxitin Oxacillin Vancomycin Linezolid ENTEROCOCCI 2/10 (20%) 4/10 (40%) 8/10 (80%) 4/10 (40%) 3/10 (30%) 8/10 (80%) 10/10 (100%) Klebsiella and Acinitobacter were highly sensitive to Impinem and Amikacin . The sensitivity pattern to other antibiotics is shown in (Table 5). Among gram positive organisms commonly isolated organism is enterococci showing highest sensitivity to linezolid, vancomycin, doxycyclin. (Table6) 4. DISCUSSION Our study findings are contrary to the belief. Therefore it is recommended that larger sample based studies may be taken up in which may throw better light on bacterial pathogens causing UTI and its susceptibility patterns. This prospective study was conducted over a period of six months from June 2014 to December 2014. During this period a total of 200 samples received from various clinical departments were tested for patogenic bacteria. CONS 3/6 (50%) 3/6 (50%) 4/6 (66.6%) 5/6 (83.3%) 4/6 (66.6%) 6/6 (100%) 6/6 (100%) STAPHYLOCOCCI 0/5 (Nil) 2/5 (40%) 3/5 (60%) 4/5 (80%) 3/5 (60%) 4/5 (80%) 5/5(100%) In our study the total no. of samples collected was 200 samples. Prevalence rate of isolation of urinary pathogen in our study was 77%. It is similar to the study by (Das RN etal) isolation rate was 71.6 %.( 18) In our study UTI was more in females when compared to males. This was seen with other studies by Bashir MF etal.(19) and GetenetB. etal (20) Women are more prone to UTIs than men because, in females, the urethra is much shorter and closer to the anus.(21) Several reports have indicated that females are more prone to having UTIs than males (Kolawaleetal, 2009), (22) Womens propensity to develop UTIs has also been explained on the basis of certain behavorial factors, including delays in micturation, sexual activity, the use of diaphragms and spermicides (both of which promote colonization of the periurethral area 624 Maria Sindhura John, J Biosci Tech,Vol 6(1),2015,620-626 with bacteria). Also, the length of the urethea (urethra), the dried environment surrounding the meatus, and the antibacterial properties of prostatic fluid contribute to a lower rate of infection in males. Higher proportions of patients were seen in the age group between 20-40 years followed by < 20 years age group. This was in consistent with a study by Beyene G et al (23) 12 in which 53.5% were in the age group between 19-39 years. Susan AMK (24) who concluded that most uncomplicated urinary tract infections occur in women who are sexually active, with far fewer cases occurring in older women, those who are pregnant, and in men. The incidence of UTI increases in males as the age advances because probably because of prostate enlargement and other related problems of old age. E-coli was the most common isolated organism in our study with percentage of (42.8%). This was in seen in other studies by Gupta et al (25), Moges et al(26) , Sibi et al(27). The second most common isolated pathogen was Klebsiella in our study accounting for 16.8%.This was in agreement by Khameneh etal(28). In our study E-coli was most resistant to Ampicillin and smost sensitive toNitrofurantoin (72.7%). The similar findings were seen in a study by (Bashir MF et al).Among gram positive organisms our study showed susceptibility pattern of enterococci as Linezolid> vancomycin = doxycycline Our study provides a bacteriological profile of pathogens causing UTI and its susceptibility pattern. It is recommended that it should be included in the proper diagnosis of UTI, so that timely and adequate treatment may be given to patient as delay in treatment is associated complicated UTI. ISSN: 0976-0172 Journal of Bioscience And Technology www.jbstonline.com 5. REFERENCES [1] Noor N, Ajaz M., Rasool SA., Pirzada ZA. Urinary tract infections associated with multidrug resistant enteric bacilli, characterization and genetical studies. Pak J Pharm Sci. 2004;17: 115-123. [2] Ronald AR, Pattulo MS. The natural history of urinary infection in adults. Medical clinics of North America 1991; 75:299-312 [3] Lichtenberger P, Hooton TM. Complicated urinary tract infections. 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