Candida albicans and Staphylococcus sp identify the presence of invasive and deep invasive squamous cell carcinoma induced by 4-nitroquiloline oxide
Authors: Guerra Caroline Camizotti , Neto José Nunes Carneiro , Ribeiro Daniel Araki , Tanomaru Juliane Maria Guerreiro , de Andrade Cleverton Roberto 
Published: August 2022

Abstract
Purpose: Oral squamous cell carcinoma (OSCC) sum 90 to 95 percent of oral cancers and the incidence described men aged 40 to 60 years old. The greater risk factor is the use of tobacco and alcohol, and they are described in independent or combined action. The gold standard method for OSCC diagnosis remain biopsy and histopathological examination. Vital staining, light-based detection systems, auto fluorescence, cytological techniques, molecular analysis, imaging diagnostic, onco-chips were described to obtain early diagnosis. The aim of this study was verifying the correlation of pool of microorganisms, a pool of bacteria, Streptococcus sp, Enterococcus faecalis, and Candida albicans and the presence of OSCC in 4-NQO cancer-induced rats.Methods: The rats’ saliva was collected one day before the sacrifice of animals. The tongue was gently scraped with a swab while the mouse was immobilized. An Eppendorf vial containing 1 mL of PBS was vortex for 2 minutes and a decimal serial dilution was realized to be plated in selective culture media for streptococcus sp (agar mitis salivarius, staphylococcus (hypertonic egg yolg agar), and for enterococcus faecalis and Candida albicans (saboroud dextrose). After microorganisms’ growth, the number of colony-forming units (CFU/mL) was identified and a log10 transformation was realized to perform statistical analysis. Results: Analysis of the histological findings was performed, and the results were described as induced and healthy. The non-statistical difference was verified in Candida albicans, Staphylococcus sp, and Streptococcus sp in 4-NQO induced animals. Correlation analysis was performed using acanthosis, papillary hyperplasia, dysplasia (mild, moderate, and multiple), papilloma, squamous cell carcinoma (SCC [in situ, microinvasive, invasive, and deep invasive]), inflammatory infiltrate (intensity [low, moderate and intense] and type [acute, mixed and chronic]), SCC inflammatory infiltrate, presence of superficial epithelial microorganism and presence of connective microorganism, the pool of microorganisms, the pool of bacteria, data on Streptococcus sp, Enterococcus faecalis and Candida albicans. A positive correlation between the pool of microorganisms with the pool of bacteria (0.98), Candida albicans (0.81), Enterococcus faecalis (0.69), Staphylococcus sp (0.67), invasive-SCC (0.87), inflammatory intensity (0.82) was identified. The bacterium pool shows positive correlation between invasive-SCC (0.78), inflammatory intensity (0.74), Enterococcus faecalis (0.74), Staphylococcus sp (0.71), Candida albicans (0.69). The Streptococcus sp shows a positive correlation between Staphylococcus sp (0.93). Conclusion:The Candida albicans show a positive correlation between invasive-SCC (0.78) and, inflammatory intensity (0.69). The Staphylococcus sp shows a positive correlation between invasive-SCC (0.70). The pool of microorganisms was correlated to SCC superficial microorganisms (1.00), an acute type of inflammatory infiltrate(1.00), the pool of bacteria (0.79), Candida albicans (0.72), and Enterococcus faecalis (0.79). In the same way, the pool of bacteria showed a correlation with Enterococcus faecalis (0.78). The dysplasia showed a correlation to moderate dysplasia (0.83). The moderate dysplasia showed a statistical correlation with Staphylococcus sp.