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 Table of Contents  
ORIGINAL ARTICLE
Year : 2020  |  Volume : 4  |  Issue : 1  |  Page : 65-68

To study the isolation and identification of fungi from oral cancer after radiotherapy


1 Department of Microbiology, MGS University, Bikaner, Rajasthan, India
2 Department of Emergency Medicine, S P Medical College, Bikaner, Rajasthan, India
3 Department of Medicine, S P Medical College, Bikaner, Rajasthan, India

Date of Submission26-Dec-2019
Date of Acceptance05-Jan-2020
Date of Web Publication17-Mar-2020

Correspondence Address:
Miss. Vibha Khatri
Department of Microbiology, MGS University, B-3 Shastri Nagar, Bikaner - 334 001, Rajasthan
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/bbrj.bbrj_166_19

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  Abstract 


Background: Cancer, known medically as a malignant neoplasm, is a broad group of diseases involving unregulated cell growth. In oral cavity infection, the oral microflora may be subsequently replaced by potentially pathogenic microorganisms such as candela species (from 72% to 92%). Hence, because of a weakened line of defense in oral cancer patients, the present prospective cohort study was carried out with the aim of isolation and identification of fungal colonization from oral cavity from radiotherapy. Radiotherapy and both radio-chemotheray treated patients. Methods: The proposed study was carried out on clinical samples in the Department of Microbiology in S. P. Medical College, Bikaner, Rajasthan. Isolates from clinical samples were collected from Acharya Tulsi Regional Cancer Hospital, Bikaner, Rajasthan. Samples of the lesion from the tongue and mouth were obtained with a sterile cotton swab. The sample was cultured on Sabouraud's dextrose agar and typical yeast colonies were determined after 72 h. After, staining isolates were subjected to biochemical identification. Results: A total of fifty isolates were taken for the study. Out of these fifty isolates, 45 (90%) were male, whereas 5 (10%) were female. In this study, about 90% of oral and pharyngeal cancer in men and around 10% in women can be estimated. Out of fifty, the highest isolates obtained were Candida albicans (15, 30%), followed by Candida glabrata (11, 22%) and Candida tropicalis (8, 16%), and the lowest number was of Candida krusei (6, 12%). In this purposed study, out of fifty patients, C. albicans can be isolated in 15 patients that is a higher value. Conclusion: C. albicans is the most commonly implicated organism in the mouth after radiotherapy and radio chemotherapy. It is clear that C. albicans is the most commonly found fungi to cause secondary infection.

Keywords: Cavity, flora, fungi, oral, radiotherapy


How to cite this article:
Khatri V, Kumar H, Singh VB, Meghwanshi GK. To study the isolation and identification of fungi from oral cancer after radiotherapy. Biomed Biotechnol Res J 2020;4:65-8

How to cite this URL:
Khatri V, Kumar H, Singh VB, Meghwanshi GK. To study the isolation and identification of fungi from oral cancer after radiotherapy. Biomed Biotechnol Res J [serial online] 2020 [cited 2020 Mar 29];4:65-8. Available from: http://www.bmbtrj.org/text.asp?2020/4/1/65/280877




  Introduction Top


Cancer, known medically as a malignant neoplasm, is a broad group of diseases involving unregulated cell growth. There is no definition that describes all the cancers. They are large family of diseases which form a subset of neoplasm, which show some features that suggest of malignancy. Oral cancer, subtypes of head-and-neck cancer, is any cancerous tissue growth located in the oral cavity.[1]

In oral cavity infection, the oral microflora may be subsequently replaced by potentially pathogenic microorganisms such as candela species (from 72% to 92%); Candida carriage was reported commonly in cancer patients, with Candida albicans being the predominant species in patients who undergo radiotherapy for head and neck.[2],[3],[4],[5],[6]

Hence, because of a weakened line of defense in oral cancer patients, the present prospective cohort study was carried out with the aim of isolation and identification of fungal colonization from oral cavity from radiotherapy. Radiotherapy and both radio-chemotheray treated patients.

Certain fungal organisms, notably C. albicans, are commensally inhabitants of the oral cavity in a large proportion of individuals. Under normal conditions, these fungal organisms of the normal oral flora do not cause disease. However, changes in the oral and/or systemic environment can result in an overgrowth of these fungal spp., leading to clinical oral fungal infection. These changes include:[7]

  1. Immunosuppression – induced by drugs or disease
  2. Imbalance in the oral flora – secondary to antibiotic therapy
  3. Hyposalivation – induce by drugs, disease, or radiation therapy
  4. Local tissue damage – mucositis secondary to chemotherapy and/or radiation therapy.


Oral candidiasis accounts for the vast majority of oral fungal infections and can have a number of clinical presentation, including [8]

  1. Pseudomembranous candidiasis (thrush): present as white curd-like pseudomembranes, which can be removed with some pressure, leading behind an erythematous mucosa
  2. Chronic hyperplastic candidiasis: presents as a hyperkeratosis white patch, with or without hyperplasia of epithelial tissue, which cannot be removed by scraping
  3. Erythematous candidiasis: presents as erythema, fissuring, and crusting of the commissures (angles) of the lips.


Oral candidiasis is candidiasis that occurs in the mouth. That is, oral candidiasis is a mycosis (yeast) fungal infections of Candida spp. on the mucosal membranes of the mouth.

C. albicans is the most commonly implicated organism in this condition. C. albicans is carried in the mouth of about 0% of the world's population as a normal component of the oral microbiota.[9]

Three main clinical appearances of candidiasis are generally recognized: pseudomembranous, erythematous (atrophic), and hyperplastic.[10] Most often affected individuals display one clear type or another, but sometimes, there can be more than one clinical variant in the same person.[11]


  Methods Top


The proposed study was carried out on clinical samples in the department of microbiology in S. P. Medical College, Bikaner, Rajasthan. Isolates from clinical samples were collected from Acharya Tulsi Regional Cancer Hospital, Bikaner, Rajasthan. Samples of the lesion from the tongue and mouth were obtained with a sterile cotton swab. The sample was cultured on Sabouraud's dextrose agar (SDA), and typical yeast colonies were determined after 72 h [Table 1].
Table 1: Strains on Sabouraud's dextrose agar

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After, staining isolates were subjected to biochemical identification.

Procedure

  • With an inoculation needle, half of the single colony is transferred in 0.5 ml of serum to the test tube
  • Incubate at 37°C for a maximum of 1½ h.


Results

Strains of C. albicans produce germ tubes from their yeast cells constitute a positive test.

Carbohydrate fermentation test [Table 2]
Table 2: Indicator on chromoagar media in Carbohydrate fermentation test

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Media and reagents

  1. Indicator broth medium


  2. Peptone 1.0 g

    Sodium chloride 0.5 g

    Beef extract 0.5 g

    Bromocresol purple 0.10 ml

  3. Carbohydrate solution 0.3 ml 20%


Procedure

  • With a sterile loop, take few isolated colonies from SDA plate in 2.0 ml sterile saline in a test tube
  • Durham's tube is placed in an inverted position in screw-capped test tubes filled with 5–6 ml indicator broth medium
  • Take 0.2 ml of culture suspension in indicator broth medium with sugars
  • Incubate the tubes at 30°C or 37°C for 2–10 days
  • Result: the presence of bubbles in Durham's tube indicates the fermentation of sugars.



  Results Top


The proposed study was carried out on oral swab samples obtained from Cancer Hospital and Research Center, Bikaner, Rajasthan. The samples were processed according to the routine diagnostic guidelines.

In our study, 45 (90%) were male and 5 (10%) were female from the total isolates patients [Table 3] and [Graph 1].
Table 3: Distribution of patients according to sex

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In our study, out of fifty patients, 10 (20%) patients are betel chewing, 39 (78%) patients were smoker (bidi/cigarette), 4 (8%) patients were tobacco chewing, and rest (12, 24%) patients were drinker [Table 4] and [Graph 2].
Table 4: Distribution of patients according to carcinogenic factor

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In our study, out of fifty, the highest isolates obtained were C. albicans (15, 30%), followed by Candida glabrata (11, 22%) and Candida tropicalis (8, 16%), and the lowest number was of C. krusei (6, 12%) [Table 5], [Graph 3] and [Figure 1], [Figure 2], [Figure 3].
Table 5: Distribution of patients according to occurrence of organism in sample

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Figure 1: Growth appear on Sabouraud's dextrose agar medium

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Figure 2: Growth on candid chrome

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Figure 3: Germ tube shown by Candida albicans

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  Discussion Top


According to the International Agency for Research on Cancer (IARC) carcinogenic cause to human including tobacco, alcohol, chewing betel etc.[12]

The IARC classified tobacco smoking as cause of oral cavity, tonsil pharynx, and nasopharynx cancer.[12]

Smoking tobacco causes 70% of oral and pharyngeal cancer in men and around 55% in women in the UK.[13]

In this study, about 90% of oral and pharyngeal cancer in men and around 10% in women can be estimated.

The proposed study was conducted on fifty patients, in which 78% of patients would be smoker, and 8% of patients would be tobacco chewer.

The IARC classifies alcohol as a cause of oral cavity tonsil and pharynx cancer.[12],[13] According to the IARC, drinking causes around 37% of oral and pharyngeal cancer in the UK.[14]

Among this study, out of fifty patients, 12 were found to be drinker and around 24% of patients are drinker.

A study shows that oral candidiasis is candidiasis that occurs in the mouth. That is, oral candidiasis is a mycosis (yeast/fungal infection) of Candida species on the mucous membrane of the mouth.[9]

C. albicans is the most commonly implicated organism in this condition. C. albicans is carried in the mouth of about 50% of the world's population as a normal component of the oral microbiota.[9]

In this proposed study, out of fifty patients, C. albicans can be isolated in 15 patients that is a higher value. Other species of Candida would be isolated from the patient. By this, it is clear that C. albicans is most commonly found fungi to cause secondary infection.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Mandal A. Available from: http://training seer.cancer.gov/disease/.categorized classification.html. [Last accessed on 2008 Sep 20].  Back to cited text no. 1
    
2.
Raber-Durlacher JE, Barasch A, Person DE, Lalla RV, Schubert MM, Fibbe WE. Oral complication and management consideration in patients treated with high dose chemotherapy. Support Care Ther 2004;1:219-29.  Back to cited text no. 2
    
3.
Leng WK, Dassanayake RS, Yau JY, Jin LJ, Yam WC, Samaranayake LP. Oral colonization, phenotypic, and genotypic profile of Candida species in irradiated, dentate, xerostomic, nasopharyngeal carcinoma survivors. J Clin Microbial 2000;38:2219-26.  Back to cited text no. 3
    
4.
Al-Abeid HN, Abu-Elteen KH, El Karmi AZ, HamadMA. Isolation and characterization of Candida species in Jordanian cancer patients colon prevalence, pathogenic determinants and anti fungal sensitivity. Jpn J Infect Dis 2004;57:279-84.  Back to cited text no. 4
    
5.
Jhan BC, Freire AR. Oral complication of radio therapy in the head and neck. Rev Bras Otorrinolaringol 2006;72:704-8.  Back to cited text no. 5
    
6.
Jhan BC, Franca EC, Oliveira RR, Santos VR, Kowalski LP, Silva AA, et al. Candida oral colonization and infection in Brazilian patients undergoing head and neck radio therapy: A pilot study. Oral Surg Oral Med Oral Pathol Oral Radiol Endodontol 2007;103:355-8.  Back to cited text no. 6
    
7.
Rhodus NL. Treatment of oral candidiasis. Northwest Dent 2012;91:32-3.  Back to cited text no. 7
    
8.
Soames JV, Southam JC, JV. Oral Pathology. 3rd ed. Oxford: Oxford University Press; 1999. p. 147,193-200.  Back to cited text no. 8
    
9.
Coulthard P, Horner K, Sloan P, Theaker E. Master Dentistry: Oral and Maxillofacial Surgery, Radiology, Pathology and Oral Medicine. 2nd ed., Vol. 1. Edinburgh: Churchill Livingstone/Elsevier; 2008. p. 180, 181,194-5.  Back to cited text no. 9
    
10.
Park KK, Brodell RT, Helms SE. Angular cheilitis, part 1: Local etiologies. Cutis 2011;87:289-95.  Back to cited text no. 10
    
11.
Salerno C, Pascale M, Contaldo M, Esposito V, Busciolano M, Milillo L, et al. Candida-associated denture stomatitis. Med Oral Patol Oral Cir Bucal 2011;16:e139-43.  Back to cited text no. 11
    
12.
Boffetta P, Hecht S, Gray N, Gupta P, Straif K. Smokeless tobacco and cancer. Lacet Oncol 2008;9:667-75.  Back to cited text no. 12
    
13.
Parkin DM. Cancers attributable to consumption of alcohol in the UK in 2010. Br J Cancer 2011;105:S14-8.  Back to cited text no. 13
    
14.
Grulich AE, van Leeuwen MT, Falster MO, Vajdic CM. Incidence of cancers in people with HIV. AIDS compared with immunosuppressant transplant receptions a meta analysis. Lancet 2007;9581:59-67.  Back to cited text no. 14
    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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