The Effects of Cranberry Juice on the(take out, so it flows better) Urinary Tract Infection Causing Bacteria Known as Escherichia coli.

Shirin Moftakhar and Assal Parsa

Department of Biological Sciences

Saddleback College

Mission Viejo CA 92692

Previous clinical research has found that the consumption of cranberry products can inhibit urinary tract infection by preventing the adhesion of Escherichia coli to uroepithelial cells. The two main reasons(compounds) theorized to contribute to the inhibition of UTI are the(take out) fructose and an unknown polymeric compound found in cranberries. Investigators are interested in testing the effects of cranberry juice on the urinary tract infection causing bacteria, E. coli. Two experimental groups were tested,; one was the cranberry juice and the other fructose solution. Thirty samples of nutrient broth were inoculated with E. coli and spread on thirty Petri dishes of nutrient agar. They were divided in three groups of ten with each group containing chads dipped in cranberry juice for one group, fructose for the other, and deionized water for the last. After they were incubated for 72 hours at 37°C, the zones of inhibitions were measured. <An> ANOVA( single factor or what?) yielded a change(?, do u mean a significant difference between initial amt & final amt of E. coli) and a Bonferroni correction test was ran(run). The results of the <Bonferroni> post hoc test indicated that cranberry juice and fructose repectively (SP, respectively) had a significant effect on the inhibition of E. coli (p=0.0167).

Introduction

According to National Institutes of Health (NIH), the second most common type of infection affecting women, the elderly, and infants are(is) urinary tract infections (UTIs) meaning(studies have shown that) one in three will have a UTI at least once in her(their) lifetime. The presence of greater(more) than 100,000 units/ml of bacteria in the urine exceeding(exceeds) a(the) threshold value for significance ,and> can result in(lead to) urinary tract infection (UTI). Infection arises from bacterial growth within the usually sterile urinary tract (Guay, 2009). There are two types of UTIs: Lower UTI and Upper UTI. Lower UTIs involve only(only involve) the bladder, whereas upper UTIs involve both the bladder and the kidneys (pyelonephritis) (Jepson, 2004). Although the standard treatment for UTI is antibiotic therapy, rising clinical failure rates of trimethoprim-sulfamethoxazole due to bacterial resistance has led physicians in some areas of the country to consider prescribing fluoroquinolones as first-line treatment (Howell, 2007). Because of these resistance building abilities, Pharmaceutical companies are constantly working on the development of new antibiotics. In the meantime, alternative therapies for prevention of UTI should be taken into consideration to slow down the rate of antibiotic resistance development.

Cranberry juice (Vaccinium macrocarpon) has been used widely(widely used) for many years to prevent and treat urinary tract infections. There is positive clinical evidence that the consumption of cranberry juice can decrease the number of symptomatic UTI’s based on two randomized double-blind studies on women over a 12-month period (Avorn et al.>(,year? 1994) and Kontiokari (,year? 2001)). However, there is no prior evidence <showing> that cranberry <juice> can <be used to> treat UTI once an infection is present. Cranberry juice and cranberry juice cocktail have been known to inhibit the adherence of bacteria such as E. coli to host tissue (Zafriri, 1989). Another study also suggests that benefits of cranberry were due to its acidity (Liu, 2008). Cranberries contain two compounds which inhibit adherence – fructose and a polymeric compound of unknown nature (Jepson, 2004). Although many juices contain fructose, only cranberries and blueberries contain the <unknown> polymeric compound. Cranberry juice will create an acidic state by lowering the pH; this acidic environment will kill and inhibit the growth of bacteria. The purpose of this study is to determine if cranberry juice, with its combination of fructose and the unknown polymeric compound, or <just> fructose <alone> do(take out, will) inhibit the growth of UTI by testing it on E. coli growth.

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Methods and Materials

Nutrient Agar for culturing Escherichia coli was prepared on 4 November 2009 in the biology lab at Saddleback College, Mission Viejo, CA. 15 ml(mL’s) of the nutrient agar was plated in 30 Petri dishes using the aseptic technique. On 5 November 2009, 0.25 ml(mL’s) of E. coli was lawn spread onto all 30 dishes. 20 ml of fructose with a 0.60 M concentration was prepared. 20 ml of Kirkland Signature cranberry juice cocktail and deionized water were poured into two 100 ml beakers. One hundred and eighty chads were made from Wattmans filter paper and autoclaved for 2 hours. Two chads dipped in DI water were placed on each of the first 10 Petri dishes as the control group which makes n=20. Two chads dipped in a previously prepared fructose solution were placed on the each of the 10 petri dishes in the second group. On each of the 10 Petri dishes in the last group, the chads were dipped in cranberry juice cocktail. All 30 Petri dishes were placed in the incubator for 72 hours at 37°C. On 11 November 2009, the Petri dishes were inspected and the zone of inhibition for each dish was measured in centimeters. All data were transferred to Microsoft Excel 2007 where further statistical manipulations(calculations) were preformed. Since a change(?, significant diff.)was found when <an> ANOVA( single factor or what?) <Test> was run; a Bonferroni correction test was done to compare the results within the groups.

Results

The DI water <Petri dishes> had E. coli growth on top of the chads therefore the zone of inhibition is(was) zero, while the fructose and cranberry juice had no(zero) growth on the chads with a ring of no(absent of) E. coli around it. Then (There) was a significant difference between all three groups(p-value’s?). The greatest difference <in inhibition zones> was between the DI water <with zero cm> and the cranberry juice <with a mean of 1.265cm>, while the least(smallest) <difference> was between the fructose<with a mean of 1cm> and cranberry juice <with a mean of 1.265cm>.

Figure 1. The Cranberry Juice had the highest adhesion rate with a mean value of 1.265 cm. The Fructose also had some(look’s like a fairly close amt compared to the cranberry, this is where the error bars will help show that they r actually significantly diff.) adhesion with a mean of 1 cm. The DI water had an adhesions rate of 0(zero). The p-value found (for what test?) for the groups was 0.0167. (where are the SEM error bar’s on the cranberry and Fructose columns?)

Discussion

Based on our results, it can be concluded that cranberry juice does inhibit E. coli growth thus inhibiting/preventing urinary tract infection (UTI). The mean difference between the control group and the experimental groups was at least 1 cm. While cranberry juice and fructose did have signs of inhibition, the chads in the deionized water were completely covered in E. coli. There was more of a( a greater) difference between cranberry juice and the DI water then the fructose and the DI water. This suggests that although cranberry juice <combination> was the main prohibitor( had the greatest inhibitory effect), the fructose in the cranberry juice( alone) has inhibitory factors as well. Zafiri et al. (1989) found that the fructose in cranberry juice and cocktails inhibits the adherence of type 1 fimbriae of E. coli to the urinary tract epithelial cells. He also found that there are two types of inhibitors in cranberry juice, a nondialyzable and a dialyzable one. It was brought to his attention that the main inhibitor was the dialyzable one. The finding that both cranberry juice cocktail and 5% fructose inhibited yeast agglutination by purified type 1 fimbriae proves that the fimbriae are the target of inhibitory action. The P fimbriated E.coli, was also inhibited by the cocktail. The reason they were only inhibited by the cranberry juice, was that the inhibitor was nondialyzable, suggesting its high molecular weight.

In Ahuja’s study it was seen that the growth of E. coli in cranberry rich agar inhibited the attachment of the p receptor specific beads to the epithelial tissue. Until the chemical nature of the inhibitor is discovered, it is not possible to speculate on the methods by which it binds to P fimbriated E. coli surfaces. Therefore the possibility that the possibility that(repeat) the nondialyzable constituents may inhibit adherence mediated by adhesions other than P fimbriae cannot be ignored. As stated in the introduction, it was suggested that cranberry juice cocktail may primarily act as a preventive agent in urinary tract infection. So the possibility that the fructose is abrosbed (absorbed) in the alimentary tract should be considered. Since cranberry juice has fructose levels higher than the required amount necessary for type 1 fimbriated E. coli inhibition, it is conceivable that the inhibitory levels of sugar are determined in the colon where most of the E. coli is present. A study by Guay showed that antibiotic resistant P fimbriated E. coli lost the ability to adhere to bladder cell receptors in humans who had consumed 240 ml of cranberry juice cocktail. This suggests that consuming cranberry juice on a daily basis can not only prevent UTI, but also slow the pace of antibiotic resistance development by reducing the need for antibiotics. This strategy can be very helpful considering the increasing antibiotic resistance rates due o(to) over-use of antibiotics.

Literature Cited

Ahuja S, Kaack B, Roberts J: Loss of fimbrial adhesion with the addition of Vaccinum macrocarpon to the growth medium of Pfimbriated Escherichia coli. J Urol 1998;159:559–562.

Avorn, J., Monane, M., Gurwitz, J. H., Glynn, R. J. et al.,

Reduction of bacteriuria and pyuria after ingestion of cranberry juice, JAMA 1994, 271, 751–754.

Guay, David R.P. (2009). Cranberry and Urinary Tract Infections. Drugs, 69 (7). 775-807.

Gupta, K., Hooton, T. M., Stamm, W. E., Increasing antimicrobial resistance and the management of uncomplicated community-acquired urinary tract infections, Ann. Intern. Med. 2001, 135, 41–50.

Howell, Amy B. (2007). Bioactive compounds in cranberries and their role in prevention of urinary tract infections. Molecular Nutrition & Food Research, 51. 732-737.

Jepson, RG., Mihaljevic, L., and Craig, J. (2004). Cranberries for preventing urinary tract infections. The Cochrane Library, 1. 1-19.

Kontiokari, T., Sundqvist, K., Nuutinen, M., Pokka, T. et al., Randomised trial of cranberry-lingonberry juice and Lactobacillus GG drink for the prevention of urinary tract infections in women, Brit. Med. J. 2001, 322, 1571 –1573.

Liu, Yatao, Gallardo-Moreno, Amparo M., Pinzon-Arango, Paola A., Reynolds, Yorke, Rodriguez, Guadalupe, Camesano, Terri A. (2008). Cranberry changes the physicochemical surface properties of E. coli and adhesion with uroepithelial cells. Colloids and Surfaces B: Biointerfaces, 65. 35-42.

Zafriri, Dina, Ofek, Itzhak, Adar, Rivka, Pocino, Marisol, and Sharon, Nathan (1989). Inhibitory Activity of Cranberry Juice on Adherence of Type 1 and Type P Fimbriated Escherichia coli to Eucaryotic Cells. Antimicrobial Agents and Chemotherapy, 33. 92-98.


Review Form

Department of Biological Sciences

Saddleback College, Mission Viejo, CA 92692

Author (s):_Shirin Moftakhar and Assal Parsa _

Title:_The Effects of Cranberry Juice on the Urinary Tract Infection Causing Bacteria Known as Escherichia coli.

Summary

Summarize the paper succinctly and dispassionately. Do not criticize here, just show that you understood the paper.

The researchers tested to see if there was an inhibitory effect on the growth to E. coli by introducing chads soaked in 3 different solutions: cranberry cocktail, fructose with a 0.60 M concentration, and Deionized water. They incubated the 30 Petri dishes samples, 10 in each group for 72 hours, and then measured the zones of inhibition around the different chads and compared the averages for each group using one of the ANOVA test’s and also ran a Bonferroni correction test and found there to be a significant difference between the groups.

General Comments

Generally explain the paper’s strengths and weaknesses and whether they are serious, or important to our current state of knowledge.

The paper had a lot of minor grammar mistakes and flow hang ups, which most would have probably been caught if it was read over a few times, but overall it was good has a good concise results section and a little adjusting to make the results a bit more clear where the differences were but the mat and methods were very though.

Technical Criticism

Review technical issues, organization and clarity. Provide a table of typographical errors, grammatical errors, and minor textual problems. It's not the reviewer's job to copy Edit the paper, mark the manuscript.

This paper was a rough draft