Journal of Biotechnology & Biomaterials
Open Access

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.

Figure 1: Growth of Bacteria on MacConkey agar.

Figure 2: Growth of Bacteria on Nutrient agar.

Figure 3: Growths of Klebsiella sps. on MacConkey agar.

Figure 4: Growths of Candida sps. on Sabouraud Dextrose agar medium.

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)

Figure 5: Result of Indole production.

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)

Figure 6: Result of Methyl red test.

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).

Figure 7: Voges Proskauer test.

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).

Figure 8: Oxidase test.

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)

Figure 9: Catalase test.

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)

Figure 10: Urease test.

After final identification of the bacteria drugs sensitivity pattern using the following drugs (Table 3).

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).

Figure 11: Disc diffusion method result.

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)

Table 1: Shows 10 cases of pregnant and 05 cases were normal women.

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 &

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%).

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.

1. Abrutyn E, Boscia A, Kaye D (1988) The treatment of asymptomatic bacteriuria in the elderly. J Am Geriatr Soc 36: 473-475.

                Google Scholar, Crossref

2. Al-Sibai M, Saha A, Rasheed P (1989) Socio-biological correlates of bacteriuria in Saudi pregnant women. Public Health 103: 113-121.

Indexed at, Google Scholar, Crossref

3. Amiri F, Rooshan M, Ahmady M, Soliamani M (2009) Hygiene practices and sexual activityassociated with urinary tract infection in pregnant women. East Mediterr Health J 15: 105-108.

Indexed at, Google Scholar

4. Anick B, Fabiano S, Ema F, Sylvie P (2011) Urinary Tract Infections during Pregnancy. InTech 7: 113-132.

Indexed at, Google Scholar, Crossref

5. 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.

Indexed at, Google Scholar    

6. Bauer A, Kriby W, Turck M (1966) Antibiotic’s susceptibility testing by standardized single disc method. Am J Clin Pathol 45: 493-496.

Indexed at, Google Scholar, Crossref

7. Tamalli M, Bioprabhu S, Alghazal MA (2013) Urinary tract infection during pregnancy at Al-khoms, Libya. Int J Med Med Sci 3: 455-459.

Indexed at, Google Scholar