Isolation and Identification of Microbial Agents Causing Urinary Tract Infection (UTI) in Pregnant Women
Received: 03-Jun-2022 / Manuscript No. jbtbm-22-66512 / Editor assigned: 06-Jun-2022 / PreQC No. jbtbm-22-66512(PQ) / Reviewed: 20-Jun-2022 / QC No. jbtbm-22-66512 / Revised: 22-Jun-2022 / Manuscript No. jbtbm-22-66512(R) / Accepted Date: 26-Jun-2022 / Published Date: 27-Jun-2022 DOI: 10.4172/2155-952X.1000278
Abstract
Urinary tract infection (UTI) is linked to a high rate of morbidity and long-term consequences in children, including renal scarring, hypertension, and chronic renal failure. UTI manifests with vague symptoms in paediatric patients, making the diagnosis more difficult. As a result, the high prevalence of untreated and inadequately managed UTI in children is a source of clinical and societal concern.
Aims of that project are Isolation & Identification of bacterial flora in Urine samples from pregnant & non pregnant women both in study & Control group. Its drugs sensitivity testing and compare our data with other studies. This study was conducted in Helix Biogenesis Pvt. Ltd. Noida & samples were collected from Different Private Hospital & Nursing Home, Noida. In this study 15 cases were selected (10 cases Pregnant women in study group & 05 cases non-pregnant women in control group) of Urine sample from different age groups.
Keywords: UTI; Women; Isolation; Identification; Microbes
Keywords
UTI; Women; Isolation; Identification; Microbes
Introduction
Urinary tract infections (UTIs) are one of the most common issues that family physicians face. UTIs during pregnancy are one of the most frequent health issues globally, particularly in underdeveloped nations (Abrutyn E et al., 1998) [1]. Urinary tract infections (UTI), which are caused by the presence and proliferation of germs in the urinary system, are one of the most frequent bacterial illnesses in humans and in pregnancy; they can affect the lower urinary tract or the bladder. UTI is reported in 20% of pregnant women and is the most prevalent reason for admission to obstetrical wards [2].
In an asymptomatic patient, UTI is defined as the presence of at least 100,000 organisms per millilitre of urine, or more than 100 organisms/ml. of urine with associated pyuria (>5 WBCs/HPF) in a symptomatic patient. A positive culture for auro pathogen should support a diagnosis of UTI, especially in healthy individuals [3]. Many physiological, anatomical, and interpersonal variables all play a role in this issue throughout maternity [4]. Urinary tract infection during pregnancy is a major cause of maternal and perinatal morbidity. Urinary tract infection during pregnancy is linked to abortion, small birth size, maternal anaemia, hypertension, premature labour, phlebitis, thrombosis, and persistent pyelonephritis [5].
Gram-negative enteric bacilli are the most common pathogens that cause UTI in children. The most prevalent etiological agents are Escherichia coli and Klebsiella. However, in recent years, Enterococci, yeasts, and Staphylococcus aureus have emerged as significant agents, and many of them are resistant to various antibiotics 3, 4. Because urine includes urea, prolonged infection with urea splitting organisms such as Proteus and Klebsiella species may develop to urinary calculi in patients1. As a result, early detection of UTI is required to prevent illness and death, as well as the treatment load on patients. This study is being conducted in response to the aforementioned issue.
Material and Method
Hanging drop preparation: Firstly take a clean groove glass slide and a clean cover slip after that with the help of sterile inoculating loop take a full of liquid culture and put it in the centre of the cover slip. Then carefully lift the slide along with drop hanging from the under surface of the cover slip. Examine the preparation under the microscope. The first focus in 10X then turn to high power objective and observe the motility of the bacteria at the edge.
Inoculation of the urine samples: Inoculation of the Urine sample on different medium including MacConkey’s Agar, Nutrient Agar Medium, Savoured Dextrose Agar Medium. After inoculation culture plate kept in incubator at 37ºC for 24-72 hrs. After incubation time were completed and observe the colonies morphology (Figures 1-4), respectively. Gram’s staining, Motility and performed the biochemical test for final identification. After that perform the Sensitivity test.
Biochemical identifications:
Indole production In 5 ml of peptone water 1 ml of the inoculum was inoculated and incubated at 37ºC for 24 hours. After incubation 5 drops of Kovac’s reagent was added to the surface.
Result - reddening of medium (Figure 5)
Methyl Red test 1 ml of inoculums were inoculated into 5 ml of Glucose Phosphate broth and incubated at 37°C for 48 hours. 5 drops of Methyl red indicator were applied to the surface after incubation. To disseminate the methyl red, the tube was rolled between the palms of the hands.
Result – bright red medium (Figure 6)
Voges-Proskauer test In 5ml of Glucose Phosphate broth, 1 ml of inoculums was inoculated and incubated at 37ºC for 48 hours. After incubation 0.6 ml of alpha-naphthol was added and shacked. Then 0.2 ml of 40% potassium hydroxide was added, mixed gently and left 10-15 minutes for colour development.
Result – pink colour slowly develops in the medium (Figure 7).
Oxidase test A piece of filter paper was placed on a cleaned microscopic slide. Then added 2-3 drops of oxidase reagent was placed on the filter paper. The isolated colony was smeared with a loop on the filter paper. The presence of a dark purple colour was observed. The reaction positive, if the smear turns purple within 10-30 second.
Result – deep purple colour on the strips or medium (Figure 8).
Catalase test A drop of 3% hydrogen peroxide was placed on to a clean microscopic slide. The isolated colony was smeared with an inoculated loop in to hydrogen-peroxide. Then observe the bubbles.
Result – release of oxygen bubbles (Figure 9)
Urease test The inoculums was inoculated in the Christensen’s Media with help of loop and incubated at 37 degree centigrade for 48 hours. If the red colour comes test is positive.
Result – red pink medium (Figure 10)
After final identification of the bacteria drugs sensitivity pattern using the following drugs (Table 3).
S. No | Name of Organism | Name of the Drugs | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
G | CF | TE | TB | NX | AK | NZ | AX | NT | AP | ||
1 | Staphylococcus sps. | 20mm | 26mm | R | R | 24mm | 28mm | 18mm | 20mm | 26mm | R |
2 | E.coli sps. | 18mm | 20mm | R | R | 20mm | - | - | - | - | - |
3 | Candida sps. | - | - | - | - | - | - | - | - | - | - |
4 | E.coli sps. | R | 20mm | 16mm | 22mm | 24mm | 18mm | 22mm | R | 22mm | 18mm |
5 | Staphylococcus sps. | 20mm | 24mm | 20mm | 22mm | 26mm | 28mm | 22mm | 18mm | 24mm | 16mm |
6 | Klebsiella sps. | R | 18mm | R | 20mm | 26mm | 18mm | R | R | 20mm | R |
7 | Streptococcus sps. | 20mm | 26mm | R | 18mm | 22mm | 28mm | 20mm | R | 24mm | 18mm |
8 | Klebsiella sps. | R | R | R | 20mm | 18mm | 22mm | R | R | 22mm | R |
9 | E.coli sps. | ||||||||||
10 | Stapylococcus sps. | 16mm | 22mm | R | R | 20mm | 26mm | 20mm | 18mm | 26mm | R |
11 | Sterile | - | - | - | - | - | - | - | - | - | - |
12 | Sterile | - | - | - | - | - | - | - | - | - | - |
13 | Staphylococcus sps. | 24mm | 18mm | 22mm | 26mm | 28mm | 20mm | 24mm | 20mm | 20mm | R |
14 | Sterile | - | - | - | - | - | - | - | - | - | - |
15 | Streptococcus sps. | R | 22mm | 26mm | R | 22mm | 26mm | 28mm | 24mm | 20mm | 18mm |
Table 3: Showed percentages of bacterial strains that were resistant to G-Gentamycin (60%), CF-Ciprofloxacin (90%), TE-Tetracycline (40%), TB-Tobramycin (60%), Nx-Norfloxacin (100%), AK-Amikacin (90%), NZ-Ofloxacin (70%), AX-Amoxicllin (50%), NT-Netilmycin (90%), and AP-Ampicillin (40%).
G-Gentamycin, CF-Ciprofloxacin, TE-Tetracycline, TBTobramycin, Nx-Norfloxacin, AK-Amikacin, NZ-Ofloxacin, AXAmoxicllin, NT-Netilmycin, AP-Ampicillin
Drug sensitivity test
Kirby- Bauer disc diffusion method: Nutrient agar plate is taken. The inoculums are prepared by touching 3-5 colonies and inoculated into sterile peptone water which is incubated at 37ºC for 4hrs. The nutrient agar plate is inoculated with the help of sterile cotton swabs. Allow the plate to dry before antibiotics discs are applied. The plates are then incubated at 37ºC for 24hrs and then the zone of inhibition is observed (Figure 11).
This methodology is adopted from Bauer A et al. [6].
After that we also performs AST to check their sensitivity result are shown in table 3.
Observations
The present study is based on the isolation and identification of Microorganisms from Pregnant & Non-Pregnant women in Study group (10 cases) & Control group (05 cases) (Table 1)
S.No | Name of patient | Age | Complains | Date of collection | Bacteria/Fungus | Media | ||
---|---|---|---|---|---|---|---|---|
Macconkey agar | Nutrient agar | Sabouraud agar | ||||||
Pregnant Women (Study Group) | ||||||||
1 | Reema | 22 | Blood in urine | 20-10-2019 | Staphylococcus sps. | + | + | - |
2 | Meena | 39 | Mucus in urine | 22-10-2019 | E.coli sps. | + | + | - |
3 | Ekta | 27 | Pain in urination | 28-10-2019 | Candida sps. | - | - | + |
4 | Devika | 32 | Pain in lower abdomen | 13-11-2019 | E.coli sps. | + | - | - |
5 | Reshma | 36 | Pain in urination | 24-11-2019 | Stapylococcus sps. | + | + | - |
6 | Sheetal | 27 | Pain in lower abdomen | 29-11-2019 | Klebsiella sps. | + | + | - |
7 | Reena | 22 | Burning in urination | 03-12-2019 | Streptococcus sps. | + | + | - |
8 | Deepa | 28 | Burning in urination | 10-12-2019 | Klebsiella sps. | + | + | - |
9 | Chahya | 33 | Blood in urine | 15-12-2019 | E.coli sps. | + | - | - |
10 | Kusum | 37 | Burning in urination | 19-12-2019 | Stapylococcus sps. | + | + | - |
Non-Pregnant Women (Control Group) | ||||||||
11 | Rita | 32 | Non pregnant woman | 19-04-2017 | Sterile | - | - | - |
12 | Prachi | 25 | Non pregnant woman | 20-04-2017 | Sterile | - | - | - |
13 | Suman | 44 | Non pregnant woman | 22-04-2017 | Staphylococcus sps. | + | - | - |
14 | Sunita | 24 | Non pregnant woman | 22-04-2017 | Sterile | - | - | - |
15 | Radha | 40 | Non pregnant woman | 25-04-2017 | Streptococcus sps. | + | - | - |
Table 1: Shows 10 cases of pregnant and 05 cases were normal women.
in different hospital in Noida region and its drugs sensitivity pattern (Table 3) from isolated bacteria. The observation as follows:-Table-1 Master table
Table-2 Isolation rate of bacterial flora in study & control group
Table- 3 Antibiotic senstivity pattern
Result
In our study Table 1 shows the distribution of study and control group. A total of 10 bacteria were isolated (Table 2) with E. coli &
S.No | Name of Bacteria | Study Group | Control Group | ||
---|---|---|---|---|---|
No. of Cases | Percentage (%) | No. of Cases | Percentage (%) | ||
1 | E.coli sps. | 3 | 30% | 0 | 0% |
2 | Klebsiella sps. | 2 | 20% | 0 | 0% |
3 | Staphylococcus sps. | 3 | 30% | 1 | 5% |
4 | Candida sps. | 1 | 10% | 0 | 0% |
5 | Streptococcus sps. | 1 | 10% | 1 | 5% |
Table 2: Reveals that presence of UTI Pathogen in control group are sterile i.e. 0% while the percentage of bacteria in study group are different that are E.coli (30%), Klebsiella (20%), Staphylococcus sp. (30%), Candida sp. (10%) Streptococcus sp. (10%).
Staphylococcus sps. (30.0%) respectively being the major organism. Next organism of importance is the Klebsiella sp. (20.0%). In control group only (5.0%) were isolate of Staphylococcus albus & Streptococcus respectively. Our finding According to Tamalli M et al. [7].Conclusion
The physiological characteristics that sensitize women to UTI during pregnancy were explored in conjunction with other characteristics such as age, sex acts, past record of UTI, & socio - economic status. As a consequence of the findings of this research, the prenatal care doctor should emphasise personal hygiene instruction to every pregnant females, particularly those from poor socioeconomic backgrounds.
Urine culture must be performed at the initial prenatal appointment, and cultures should be acquired at various trimesters because treated individuals' urine may not remain sterile throughout the pregnancy.
To minimise maternal-fetal problems, pregnant women should be treated with appropriate antibiotic medication based on sensitivity testing when bacteriuria is detected.
The most prevalent urine symptoms in pregnancy, according to this study, were irregular nullifying trend and irritative manifestation. The majority of urinary symptoms were caused by changes in the urinary system caused by pregnancy. UTI risk factors included a history of UTI, sexual activity, being from a lower socioeconomic group, and having several children. UTI in pregnancy is definitely linked to the chance of developing symptomatic pyelonephritis later in pregnancy, and it may also be linked to other maternal and foetal problems. A prenatal care inquiry should include a urine check.
Acknowledgement
We deeply acknowledge Helix Biogenesis Pvt. Ltd. Noida to give us lab to do work and Thanks to All the Co-authors to cooperate and complete the work on time.
Conflict of Interest
No author has any conflict of interest.
Ethics of approval statement
No need to take approval of ethics
Patient consent statement
All patient gives their consent.
Permission to reproduce material from other sources
No data taken from other sources
Clinical trial registration and ethical approval information
None
Author contributions
Sumit Sheoran (SS) designed the experimental work Swati Arora (SA), Nishi Aggarwal (NA), SamsonRaj R (SR) and Lopamudra Ruth (LR) do the lab works and write the final manuscript along with SS.
References
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- Assefa A, Asrat D, Woldeamanuel Y, Hiwot Y, Abdella A, et al. (2008) Bacterial profile and drug susceptibility pattern of urinary tract infection in pregnant women at Tikur Anbessa Specialized Hospital Addis Ababa, Ethiopia. Ethiop Med J 46: 227-235.
- Bauer A, Kriby W, Turck M (1966) Antibiotic’s susceptibility testing by standardized single disc method. Am J Clin Pathol 45: 493-496.
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Citation: Agarwal N, Arora S, Sheoran S, Samsonraj R, Rath L (2022) Isolation and Identification of Microbial Agents Causing Urinary Tract Infection (UTI) in Women. J Biotechnol Biomater, 12: 278. DOI: 10.4172/2155-952X.1000278
Copyright: © 2022 Agarwal N, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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