Comparative Analysis Of Oral Fungal Microbial Diversity With Human Immunodeficiency Virus Infected Patients

Human immunodeficiency virus(HIV)infection resulted in a progressive decrease in the number and function of the CD4~+T lymphocytes,making the body prone to various infections.Oral candidiasis,the most common oral infection and can be detected in the early stages of HIV infection,and was considered as the clinical guidance to find suspicious infections for oral clinicians.It is helpful to understand the occurrence and development of infections via researches on the oral microorganisms of HIV-infected patients.At present,the researches on oral microorganisms of HIV-infected patients are mainly focusing on oral bacteria.There are many studies focusing on oral mycobiome.However,more attention was focused on the oral mycobiome which can be cultured in the laboratory.The role of many non-cultured oral mycobiome in HIV-infected patients is yet not clear.Objective:This study was designed to analyze oral fungal microbial diversity of HIV-uninfected individuals and HIV-infected patients who enrolled again after 6 months antiretroviral therapy(ART)by using Next-Generation Sequencing(NGS),aim to explore the impact of HIV infection and ART treatment on oral fungal microorganisms,and provide a basis for the later researches of oral fungal microorganisms.Methods:30 HIV-infected patients who were diagnosed in recent one year and had not received antiviral treatment and 30 HIV-uninfected individuals were enrolled;All subjects were required to be free of systemic systemic disease and oral mucosal disease,and not used antibiotics within 1 month before sample collection.HIV-infected patients were enrolled again after 6 months antiretroviral therapy.A total of 84 oral saliva samples were collected;DNA was extracted from these samples;NGS was used to analyze the fungal microbial diversity and community structure in saliva.Results:A total of 5,021,621 reads and 5820 ZOTUs(Zero-radius Operational Taxonomic Units)were obtained,aligned with the reference of UNITE database and clustered ZOTUs at the similarity level of 100%,then we revealed 11 phyla,38 classes,86orders,179 families and 306 genera of fungal microorganisms with the platform of Illumina Mi Seq in NGS.Alpha diversity analysis revealed that the species richness and evenness of saliva exhibited much higher level in HIV-infected group compared with HIV-uninfected group and the ART-treated group(p<0.05);At the phylum level,the relative abundance of Basidiomycota,Mortierellomycota and Chytridiomycota are much higher in HIV-infected group compared with HIV-uninfected group and the ART-treated group(p<0.05);While at the genus level,the relative abundance of Candida,Mortierella,Malassezia,Penicillium,Simplicillium etc was higher in the HIV-infected group than HIV-uninfected group and the ART group;Correlation analysis revealed that Mortierella,Malassezia,Simplicillium and Chaetomium have a moderate degree positive correlation with viral load(VL),while Thyrostroma and Archaeorhizomyces have a moderate negative correlation with virus load and a moderate positive correlation with CD4~+T cells.Conclusions:The results demonstrated that Mortierella,Malassezia,Penicillium,Simplicillium and Chaetomium are strongly associated with HIV infection and can be potential biomarkers of HIV infection.In addition,the species richness and evenness of saliva fungal microorganisms in HIV-infected group have a significant difference with HIV-uninfected group and ART-treated groups,indicating that HIV infection and ART treatment could affect the composition of oral fungal microbial communities.Furthermore,the correlative analysis indicated that the oral fungal microorganisms were changeable with the body’s immune status for HIV-infected patients.