Social-demographic and related background information of the study subjects.

<p>Social-demographic and related background information of the study subjects.</p

Dendrogram of genetic relationship between 15 <i>C</i>. <i>albicans</i> isolates from the S-ECC children saliva sample, based on the seven housekeeping loci.

<p>*The dendrogram was constructed by using the unweighed pair-group method. The linkage distance is indicated at the bottom.</p

Carriage of <i>C</i>. <i>albicans</i> and <i>S</i>. <i>mutans</i> and caries score in the S-ECC children and their mothers.

<p>Carriage of <i>C</i>. <i>albicans</i> and <i>S</i>. <i>mutans</i> and caries score in the S-ECC children and their mothers.</p

<i>Candida albicans</i> Carriage in Children with Severe Early Childhood Caries (S-ECC) and Maternal Relatedness

<div><p>Introduction</p><p><i>Candida albicans</i> has been detected together with <i>Streptococcus mutans</i> in high numbers in plaque-biofilm from children with early childhood caries (ECC). The goal of this study was to examine the <i>C</i>. <i>albicans</i> carriage in children with severe early childhood caries (S-ECC) and the maternal relatedness.</p><p>Methods</p><p>Subjects in this pilot cross-sectional study were recruited based on a convenient sample. DMFT(S)/<i>dmft(s)</i> caries and plaque scores were assessed during a comprehensive oral exam. Social-demographic and related background information was collected through a questionnaire. Saliva and plaque sample from all children and mother subjects were collected. <i>C</i>. <i>albicans</i> were isolated by BBL^™ CHROMagar^™ and also identified using germ tube test. <i>S</i>. <i>mutans</i> was isolated using Mitis Salivarius with Bacitracin selective medium and identified by colony morphology. Genetic relatedness was examined using restriction endonuclease analysis of the <i>C</i>. <i>albicans</i> genome using BssHII (REAG-B). Multilocus sequence typing was used to examine the clustering information of isolated <i>C</i>. <i>albicans</i>. Spot assay was performed to examine the <i>C</i>. <i>albicans</i> Caspofungin susceptibility between S-ECC children and their mothers. All statistical analyses (power analysis for sample size, Spearman’s correlation coefficient and multiple regression analyses) were implemented with SAS 9.4</p><p>Results</p><p>A total of 18 S-ECC child-mother pairs and 17 caries free child-mother pairs were enrolled in the study. Results indicated high <i>C</i>. <i>albicans</i> carriage rate in the oral cavity (saliva and plaque) of both S-ECC children and their mothers (>80%). Spearman’s correlation coefficient also indicated a significant correlation between salivary and plaque <i>C</i>. <i>albicans</i> and <i>S</i>. <i>mutans</i> carriage (p<0.01) and caries severity (p<0.05). The levels of <i>C</i>. <i>albicans</i> in the prepared saliva and plaque sample (1ml resuspension) of S-ECC children were 1.3 ± 4.5 x10⁴ cfu/ml and 1.2 ± 3.5 x10⁴ cfu/ml (~3-log higher vs. caries-free children). Among 18 child-mother pairs, >60% of them demonstrated identical <i>C</i>. <i>albicans</i> REAG-B pattern. <i>C</i>. <i>albicans</i> isolated from >65% of child-mother pairs demonstrated similar susceptibility to caspofungin in spot assay, while no caspofungin resistant strains were seen when compared with <i>C</i>. <i>albicans</i> wild-type strain SC5314. Interestingly, the regression analysis showed that factors such as antibiotic usage, birth weight, inhaler use, brushing frequency, and daycare attendance had no significant effect on the oral carriage of <i>C</i>. <i>albicans</i> in the S-ECC children.</p><p>Conclusions</p><p>Our results reveal that both the child with S-ECC and the mother were highly infected with <i>C</i>. <i>albicans</i>, while most of the strains were genetically related, suggesting that the mother might be a source for <i>C</i>. <i>albicans</i> acquisition in the oral cavity of children affected by the disease.</p></div

Examples of susceptibility of the clinical isolated <i>C</i>. <i>albicans</i> from S-ECC children and their mothers to antifungal agents casopfungin compared to <i>C</i>. <i>albicans</i> wild strain SC5314.

<p>From left to right, 10⁴, 10⁵, and 10⁶ cells were spotted on each plate and incubated at 37°C for up to 5 days. Clinical isolated <i>C</i>. <i>albicans</i> demonstrated less growth and smaller colonies than SC5314 on the YPD medium with caspofungin. However, no significant growth difference was seen between the <i>C</i>. <i>albicans</i> from S-ECC children and their mothers on the YPD medium with caspofungin.</p

Representative PFGE patterns and genetic relatedness of <i>C</i>. <i>albicans</i> obtained by restriction endonuclease analysis of genomic DNA using BssHII (REAG-B) among family members.

<p>(A) Representative PFGE patterns and genetic relatedness of <i>C</i>. <i>albicans</i> within different child-mother pairs; (B) Relatedness of <i>C</i>. <i>albicans</i> among S-ECC child-mother pairs. * Relatedness analysis was performed by BioNumerics software (Applied Maths, Austin, TX). The <i>C</i>. <i>albicans</i> isolated from the child-1 and mother-1 were genetically identical, the <i>C</i>. <i>albicans</i> isolated from the child-5 and mother-5 were genetically different.</p

Spearman’s rank correlation between <i>C</i>. <i>albicans</i>, <i>S</i>. <i>mutans and caries score among S-ECC children</i>.

<p>Spearman’s rank correlation between <i>C</i>. <i>albicans</i>, <i>S</i>. <i>mutans and caries score among S-ECC children</i>.</p