Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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Lucknow
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Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
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Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
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Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

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Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


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Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
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Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2022 | Month : August | Volume : 16 | Issue : 8 | Page : ZC22 - ZC26 Full Version

Evaluation of Antifungal Effectiveness of a Novel Probiotic Toothpaste: An In-vitro Study


Published: August 1, 2022 | DOI: https://doi.org/10.7860/JCDR/2022/55875.16752
Pavita Kataria, Harpreet Singh

1. Senior Lecturer, Department of Periodontics, Baba Jaswant Singh Dental College, Ludhiana, Punjab, India. 2. Dean, Department of Endodontics, Baba Jaswant Singh Research Institute, Patiala, Punjab, India.

Correspondence Address :
Dr. Pavita Kataria,
B-24/623, Street No. 4, Harcharan Nagar, Ludhiana, Punjab, India.
E-mail: pavita_kataria@yahoo.com

Abstract

Introduction: There is increasing public interest in probiotic healthcare products since their discovery by Élie Metchnikoff in the early 20th century. This trend is not only visible in supermarkets and dental practices, but also in the scientific world. An increasing number of clinical trials and laboratory investigations are being conducted to validate the claims made about these products with regards to their antifungal properties.

Aim: To evaluate the antifungal properties of commercially available probiotic, anticaries toothpaste, Purexa, against Candida albicans in-vitro.

Materials and Methods: An in-vitro study was carried out for a period of three months from December 2021 to February 2022 in the Department of Microbiology, Baba Jaswant Singh Dental College and Research Institute, Patiala, Punjab, India. Two wells were prepared in each blood agar plate containing lawn cultures of Candida albicans. The plates were incubated at 37°C for 24 hours. Purexa toothpaste was poured into one well as test material. The second well was filled with distilled water to act as control. A total of 10 sample plates were made. Antifungal activity was assessed by measuring the Zone of Inhibition (ZOI) in millimeters (mm) using a digital vernier caliper after 24, 48 and 72 hours, and then after one week. Increasing width of zones of inhibition indicated definite antifungal activity of Purexa toothpaste. A p-value <0.05 was considered to be statistically significant.

Results: All the sample plates showed definite zones of inhibition at 24 hours, 48 hours, 72 hours and one week intervals around the wells containing the test material, which indicated antifungal efficacy of Purexa probiotic toothpaste. No inhibition zones were seen around the wells containing distilled water. The mean width, if ZOI was the highest at day 7 (28.71 mm), followed by day 2 (26.66 mm), day 3 (26.36 mm) and least on day 1 (25.25 mm).

Conclusion: The preliminary data obtained from the present study indicates the prospective use of Purexa probiotic toothpaste in cases, where antifungal activity of toothpaste is desirable.

Keywords

Candida albicans, Dentrifices, Purexa

The oral cavity provides a habitat to a large number of microbial species which co-exist with one another as normal microbiota (1). Candida, one of the most commonly found opportunistic oral pathogens has more than 20 species (2). Candida albicans (C. albicans) is most commonly associated with oral candidal infection, in both medically compromised as well as otherwise healthy individuals (2),(3). Different pathological conditions like acute pseudomembranous thrush to more chronic forms which may persist for a longer period despite treatment can be caused by C. albicans (1). The risk of candidal infection is increased markedly due to various factors such as poor oral hygiene, diabetes mellitus, prolonged use of antibiotics, and use of dental prostheses, immunosuppression, high carbohydrates diet, or heavy smoking (4),(5).

Many antifungal medicines in the market today have several side-effects and they can also lead to development of drug resistance, causing profound effects on human health (6). These factors have led to the need to develop newer antifungal drugs with lesser side-effects. World Health Organisation (WHO) in 2001 defined probiotics as live microorganisms which when administered in adequate amounts confer health benefits on the host (7). There is increasing public interest in probiotic healthcare products since their discovery by Elie Metchnikoff in the early 20th century (8). Probiotics are known to perform several beneficial actions in the gastrointestinal tract. In relation to prevention of oral diseases, they help maintain and restore the natural saprophytic microflora against pathogen invasion, which plays the main role in development of major oral diseases (9). A large number of clinical trials are being conducted to validate the claims made about various probiotic healthcare products with regards to claims of their antimicrobial properties (10),(11),(12),(13).

Toothpastes are the most common vehicles for drug delivery in oral cavity (5). They have been used since almost 4000 years (3). Various types of abrasives were used to clean teeth till the mid 19th century. In middle-ages, the Arabs used fine sand and pumice as tooth cleaning agents. The first toothpaste was invented in 1950, by Dr. Washington Wentworth Sheffield, a dental surgeon and chemist (14). Modern dentifrices contain abrasive agents, thickening agents, humectants, flavouring agents, and antimicrobial agents. They have been formulated to contain chemotherapeutic agents to improve oral health, and to produce inhibitory action on plaque formation and on colonisation of bacteria and Candida (3),(15),(16),(17),(18). With newer toothpastes being introduced to the market regularly, there is a need for research to determine their antimicrobial and antifungal activities as well. Purexa probiotic toothpaste is novel toothpaste with probiotic activity, recently launched in Indian market. The effective ingredients of the toothpaste are bacillus coagulum, sodium monofluorophosphate, xylitol, calcium carbonate, hydrated silica, sodium citrate, and sorbitol (19). Till date, no research is available to validate the antifungal activity of the toothpaste. Thus, the present study was conducted to determine the antifungal activity of a newly introduced commercially available probiotic, anticaries toothpaste, Purexa; against C. albicans isolates using agar well-diffusion method.

Material and Methods

This in-vitro study was conducted in Department of Microbiology at the Baba Jaswant Singh Dental College and Research Institute, Patiala, Punjab, India, over a total period of three months (December 2021-February 2022). Ethical clearance for the same was obtained from Ethical Committee of Baba Jaswant Singh Research Institute (BJSRI/21/140). Purexa Probiotic toothpaste was procured and used as a test material for the present study (Table/Fig 1). Total 10 sample plates were made. Wells were prepared in blood agar plates containing lawn cultures of C. albicans. The plates were incubated at 37°C for 24 hours. Undiluted Purexa toothpaste was poured into one well as test material. The second well in each plate was filled with distilled water to act as control. Antifungal activity was assessed by measuring the ZOI in millimeters (mm) using a digital vernier caliper after 24, 48 and 72 hours, and then after one week.

Reviving the Microbial Species

Freeze-dried microbial cultures were acquired from “Microbial Type Culture Collection and Gene Bank (MTCC), Chandigarh”. Cultures of Candida albicans (MTCC-3018) were obtained in the form of freeze dried ampoules. Ampoules were cut with a diamond cutter and broken by placing them in cotton gauze. Culture of 0.5 mL was extracted from culture bottles (BACT/ALERT FA Plus Culture bottles, bioMerieux, USA) and injected into the ampoules and agitated for thorough mixing of bacterial cultures. After a resting period of 15 minutes, the microbe and culture solution was injected back into culture bottles and incubated for 48-72 hours (20). Once the incubation period was over, pure microbial growth was obtained by streaking the culture from incubated bottles onto blood agar plates (Himedia) and incubated for 24 hours. Microbial colonies were verified by gross examination of colonies and observing under microscope.

Agar Diffusion Test

Agar plates were freshly made containing Himedia Blood Agar. Fungal suspension was made by adding colonies to 0.5 mL saline using a sterile inoculating loop and adjusting the turbidity to 0.5 McFarland standards (20). A sterile swab was dipped in this suspension and used to make the lawn culture of fungi. Then, two wells, each in 4 mm depth and 5 mm diameter, were punched on the agar surface with equal distance from each other. One well was filled with 0.1 mg Purexa toothpaste and the second well was filled with distilled water, to act as control (Table/Fig 2). Agar plates were incubated at 37°C for 48 hours and the diameter of the ZOI was measured and recorded after 24, 48 and 72 hours, and then after one week.

Statistical Analysis

One way analysis of variance (F-test) was used to compare the mean value of antifungal activity using a digital vernier caliper observed at four different days i.e. at day 1, day 2, and day 3 and after one week. The Bonferroni post-hoc test was also applied for multiple inter group comparisons. Statistical Package for the Social Sciences (SPSS) version 21.0 was used for calculations. A p-value <0.05 was considered to be statistically significant.

Results

Definite ZOI was observed around the wells containing the test material (Purexa probiotic toothpaste, while no ZOI was observed around the wells containing the control material. (Table/Fig 3),(Table/Fig 4),(Table/Fig 5),(Table/Fig 6),(Table/Fig 7). The zones of inhibition observed at 24 hours, 48 hours, 72 hours and one week intervals are depicted in (Table/Fig 8).

The mean was observed to vary with time duration. The mean value came out to the highest at day 7 (28.71 mm), followed by day 2 (26.66 mm), day 3 (26.36 mm) and least on day 1 (25.25 mm). When the mean values were compared to each other using F-test, it was observed that the antifungal activity differed significantly from each other at day 1, day 2, day 3 and one week, respectively (p<0.001) (Table/Fig 9).

Intragroup multiple comparisons-Bonferroni post-hoc test: Further, the multiple comparisons between the days i.e. the groups each other has revealed that the mean width of ZOI of day 1 was statistically significant with day 2, day 3 and day 7 (p<0.001), whereas, ZOI of day 2 was significant with day 1 and day 7, while it was non-significant with day 3. Like-wise, the mean width of ZOI of day 3 was significant with day 1 and day 7, while it was non significant with day 2. The mean width of ZOI of day 7 was significant with day 1, day 2 and day 3 (Table/Fig 10).

Discussion

The present in-vitro study was performed to evaluate antifungal activity of newly introduced probiotic toothpaste, Purexa. Results indicate definite antifungal activity of Purexa probiotic toothpaste, as was evident from zones of inhibition observed during the given time period. The incidence of Candida albicans isolated from the oral cavity has been reported to be 45% in neonates, 45%-65% of healthy children, 30%-45% of healthy adults, 50%-65% of people who wear removable dentures, 65%-88% in those living in acute and long term care facilities, 90% of patients with acute leukemia undergoing chemotherapy, and 95% of patients with HIV (21),(22),(23),(24),(25),(26),(27),(28). Plaque control, a procedure that involves removal of microbial plaque biofilms and its accumulation on various hard surfaces and gingival surfaces in oral cavity, is an important step in preventing tooth decay and periodontal diseases (29). Most of the research work related to determination of antimicrobial activity of toothpastes focus on their potential to inhibit bacterial growth. There is insufficient data concerning their antifungal activity (3),(30).

A number of toothpastes containing probiotics that may be beneficial in improving oral health have been developed recently. Studies done by Chandhru TP et al., Amizic IP et al., and Adwan G et al., concluded that probiotic toothpastes can contribute in prevention of periodontal diseases (20),(31),(32). The present study was designed to evaluate antifungal activity of new commercially introduced probiotic, anticaries, Purexa. The effective ingredients of the toothpaste are 350 million CFU/gm helpful bacteria bacillus coagulum (0.02%), sodium monofluorophosphate (0.8%), xylitol (2%), calcium carbonate (42.4%), hydrated silica (3%), sodium citrate (0.25%), and sorbitol (28%) (Table/Fig 11) (19). While bacillus coagulum is known to inhibit the growth of cavity causing S. mutans (33), various clinical studies indicate that fluorides, xylitol, and probiotics demonstrate antifungal effects (1),(34),(35). Probiotics have always been considered an excellent solution for gastrointestinal issues. Their use in oral healthcare products has been investigated in recent years and is steadily gaining popularity. A large number of these investigations are conducted on dairy products that contain Probiotic strains: Lactobacillus rhamnosus, Lactobacillus router, and Bifidobacterium spp. were the most regularly used strains (33). There was a need for a probiotic strain vehicle that was stable, had a long shelf life, and could be made available and used on a regular basis by a greater segment of society (36). Purexa probiotic toothpaste, which is a therapeutic innovation, can be beneficial against C. albicans and subsequently against various dental diseases by restoration of healthy microflora. Results of the present study may provide invaluable information for dental professionals. They can recommend a dentifrice that has good inhibition properties against C. albicans for a patient, who is prone to oral candidal infections.

Limitation(s)

This testing method was more of a screening test, and may not have been able to detect the effects of chemical agents that did not diffuse through agar matrix. The findings of the present study cannot be used for evaluating its clinical effectiveness. This is because it was an in-vitro experiment and inside the oral cavity the toothpaste is likely to be diluted by saliva, to an extent that its antifungal properties may be buffered or lost due to dilution. Also, it is well understood that there is a state of balance in the oral microbiota of a healthy individual. If this balance is disturbed, the opportunistic microbes can proliferate and lead to initiation of disease processes. Therefore, a dentifrice that shows the largest inhibition zone against C. albicans may not necessarily be better than that giving a smaller inhibition zone.

Conclusion

Probiotic toothpastes are viable microbes that are known for their various health benefits when administered in adequate quantities. Probiotic toothpaste is a relatively new concept that can be useful in prevention of periodontal diseases. The present study was aimed at evaluating antifungal activity of Purexa probiotic toothpaste, which contains fluorides, xylitol, and probiotics. The paste showed substantial antifungal activity in the laboratory analysis and may be beneficial to individuals prone to oral candidiasis. However, more research needs to be done to better understand its effectiveness in the oral cavity, and whether it can be used as an alternative to conventional toothpastes in case of oral candida infection.

Acknowledgement

The authors would like to thank Dr. Aditya Khichy and Dr. Vishwas Narang for their clinical assistance; and The Baba Jaswant Singh Trust.

References

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Furlletti VF, de Cássia Mardegan R, Obando-Pereda GA, Aníbal PC, Duarte MCT, Gonçalves RB, et al. Susceptibility of Candida spp. Oral isolates for azolic antifungals and amphotericin B. Braz J Oral Sci. 2008;7(25):1543-49.
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Patel M, Shackleton JT, Coogan MM. Effect of antifungal treatment on the prevalence of yeasts in HIV-infected subjects. J Med Microbiol. 2006;55(Pt 9):1279-84. [crossref]
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Nayak A, Nayak RN, Bhat K. Antifungal activity of a toothpaste containing Ganoderma lucidum against Candida albicans- An in vitro study. J Int Oral Health. 2010;2(2):51-57. [PubMed]
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DOI and Others

DOI: 10.7860/JCDR/2022/55875.16752

Date of Submission: Feb 28, 2022
Date of Peer Review: Apr 05, 2022
Date of Acceptance: May 21, 2022
Date of Publishing: Aug 01, 2022

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? NA
• For any images presented appropriate consent has been obtained from the subjects. NA

PLAGIARISM CHECKING METHODS:
• Plagiarism X-checker: Mar 04, 2022
• Manual Googling: Apr 05, 2022
• iThenticate Software: May 20, 2022 (19%)

ETYMOLOGY: Author Origin

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