Characterization and functional prediction of the dental plaque microbiome in patients with alveolar clefts.

Introduction: Alveolar cleft (AC) is a common congenital defect in people with cleft lip and palate (CLP). Alveolar bone grafting (ABG) is typically performed during adolescence, resulting in the fissure remaining in the mouth for a longer length of time. Patients with AC have a greater rate of oral diseases such as dental caries than the normal population, and the precise characteristics of the bacterial alterations caused by AC are unknown. Methods: We recruited a total of 87 subjects and collected dental plaque samples from AC adolescents (AAP), post-operative ABG adolescents (PAP), healthy control adolescents (CAP), AC young adults (AYP), post-operative ABG young adults (PYP), and healthy control young adults (CYP). The sequencing of 16S rRNA genes was performed. Results: The microbial composition of plaque from alveolar cleft patients differed significantly from age-matched healthy controls. Linear discriminant analysis effect size (LEfSe) analysis revealed that AAP was enriched for Neisseria, Haemophilus, Fusobacterium, Rhodococcus, Aggregatibacter, Gemella, and Porphyromonas, whereas AYP was enriched for Capnocytophaga, Rhodococcus, and Actinomyces-f0332. There were phenotypic differences in facultatively anaerobic, Gram-negative, Gram-positive, and oxidative stress tolerance between the AYP group with longer alveolar cleft and the healthy control group according to Bugbase phenotypic predictions. Alveolar bone grafting did not alter the functional phenotype of alveolar cleft patients but reduced the number of differential genera between alveolar cleft patients and healthy controls at both ages. Conclusions: Our study systematically characterized the supragingival plaque microbiota of alveolar cleft patients, post-alveolar bone grafting patients, and matched healthy controls in two ages to gain a better understanding of plaque ecology and microbiology associated with alveolar clefts.

Investigating oral microbiome profiles in patients with cleft lip and palate compared with the healthy control.

BACKGROUND: Patients with cleft lip and palate (CLP) have an oronasal communication differed from the closed state in healthy individuals, leading to a unique oral microbiome. This study aimed to determine if variances in the oral microbiota persist among CLP patients who have received treatments for the closure of these fistulas compared to the microbiota of healthy individuals. METHODS: Saliva samples were collected from a cohort comprising 28 CLP patients (CLP group) and 30 healthy controls (HC group). Utilizing 16S rRNA sequencing on the Illumina NovaSeq platform, we conducted a comprehensive analysis of the diversity and composition of the oral microbiota. RESULTS: The analysis of the microbiota in the saliva samples revealed a total of 23 microbial phyla, 38 classes, 111 orders, 184 families, 327 genera and 612 species. The alpha diversity with microbial abundance and evenness indicated the significant difference between the CLP and HC groups. Principal coordinate analysis (PCoA) and the ADONIS test further supported the presence of distinct microorganisms between the two groups. The CLP group displayed elevated abundances of Neisseria, Haemophilus, Porphyromonas, and Granulicatella, as indicated by LefSe analysis. Conversely, Rothia, Veillonella, and Pauljensenia exhibited significant reductions in abundance in the CLP group. The results of the PICRUSt analysis indicated significant differences in the relative abundance of 25 KEGG pathways within the CLP group. Through Spearman correlation analysis, strong associations between Rothia, Veillonella, and Pauljensenia and 25 functional pathways linked to CLP were identified. CONCLUSION: Findings of this study offer a thorough comprehension of the microbiome profiles of CLP patients after the restoration of oronasal structure and are anticipated to present innovative concepts for the treatment of CLP.

Candida species biotypes in the oral cavity of infants and children with orofacial clefts under surgical rehabilitation.

Patients with orofacial clefts present various risk factors for oral infectious diseases, resulting from anatomical and physiological changes and those resulting from rehabilitating therapeutic interventions. The incidence of Candida species in groups of babies and children with orofacial clefts, during pre- and post-operative periods and until return to first consultation, and the profiles for antifungal sensitivity and virulence in vitro were investigated. Oral samples were collected at different times over the surgical procedures and post-surgical clinical consultation and seeded in chromogenic culture media CHROMagar Candida®. Candida biotypes were identified by accessing species-specific genomic DNA sequences by PCR techniques and electrophoretic procedures. Antifungal susceptibility testing was performed by the method of microdilution in broth using the antifungals amphotericin B (AP), nystatin (NYS) and fluconazole (FLC). SAP and PL exoenzyme activities were determined by classical microbiological methods. Some orofacial clefts occurred preferentially in male or female. Low incidence (39.1%) of oral colonization by Candida species (C. albicans, C. krusei, C. tropicalis and Candida spp.) was reported in patient admission to surgical ward, with no correlation to orofacial cleft types or surgical history. Significant reduction in frequencies of Candida and changes of species, over sampling periods, showed dynamic patterns of oral colonization: elimination, maintenance or neocolonization of the biotypes. These biotypes showed sensitivity to AP (100%), partial resistance to FLC (<10%) and variable MICs for NYS (0.125-4 µg/mL), in addition to strong exoenzyme activities, especially for SAP. Clinical and therapeutic conducts for surgical rehabilitation, anatomical and physiological characteristics of patients with orofacial clefts, and cultural behavior and regionalism of the patient population served could influence the frequencies and dynamics of oral colonization by Candida species. The data showed Candida biotypes resistant to FLC and sensitive (AP) or clinically compatible (NYS) to polyenes, especially C. albicans, in the oral cavity of patients predisposed to oral colonization and candidiases, contributing to clinical conducts in possible antifungal therapies. These biotypes were considered potentially virulent and able to partially modulate their virulence factors, especially SAP, under the conditions favored by host.

A Comparative Study of Oral Microbiota in Infants with Complete Cleft Lip and Palate or Cleft Soft Palate.

Few reports have been published on the early microbiota in infants with various types of cleft palate. We assessed the formation of the oral microbiota in infants with complete cleft lip and palate (CLP n = 30) or cleft soft palate (CSP n = 25) in the neonatal period (T1 time) and again in the gum pad stage (T2 time). Culture swabs from the tongue, palate, and/or cleft margin at T1 and T2 were taken. We analysed the prevalence of the given bacterial species (the percentage) and the proportions in which the palate and tongue were colonised by each microorganism. At T1, Streptococcus mitis (S. mitis) were the most frequently detected in subjects with CLP or CSP (63% and 60%, resp.). A significantly higher frequency of methicillin-sensitive Staphylococcus aureus (S. aureus MSSA) was observed in CLP compared to the CSP group. At T2, significantly higher percentages of S. mitis, S. aureus MSSA, Staphylococcus epidermidis, and members of the Enterobacteriaceae family were noted in CLP infants compared to the CSP. S. mitis and Streptococcus sanguinis appeared with the greatest frequency on the tongue, whereas Streptococcus salivarius was predominant on the palate. The development of the microbiota in CLP subjects was characterised by a significant increase in the prevalence of pathogenic bacteria.

Decolonisation of meticillin-resistant Staphylococcus aureus (MRSA) carriage in adopted children with cleft lip and palate.

This study aimed to determine the percentage success and to investigate influencing factors of meticillin-resistant Staphylococcus aureus (MRSA) decolonisation treatment in children with cleft lip and/or palate (CLP) who are adopted to The Netherlands. This was a historic cohort study in nine Dutch hospitals with a CLP treatment centre of children who were adopted from abroad in 2005-2012 who had CLP and MRSA carriage upon arrival in The Netherlands. A total of 55 adopted children with CLP and MRSA carriage were eligible for the study. Most children were adopted from China and had cheilognathopalatoschisis. Fourteen children were not treated for MRSA carriage, of whom six became MRSA-negative spontaneously. Forty-one children received decolonisation treatment (either topical treatment and disinfectant body wash or these combined with oral antibiotics). Overall, eighteen children [44%; 95% confidence interval (CI) 29-59%] became MRSA-negative after treatment. Treatment success was higher (56%; 95% CI 33-77%) in the group of children treated according to the Dutch guideline for treatment of MRSA carriage (odds ratio=6.1, 95% CI 4.4-26.4; p=0.017). In conclusion, MRSA decolonisation treatment in adopted children with CLP was successful in 44% of cases and the success percentage was higher in the group of children treated in accordance with the national guideline for treatment of MRSA carriage. However, given the percentage of children who turned MRSA-negative without treatment, waiting for spontaneous clearance of MRSA carriage can be advised after careful consideration of the benefits and risks of decolonisation treatment.

Investigating Oral Microbiome Profiles in Children with Cleft Lip and Palate for Prognosis of Alveolar Bone Grafting.

In this study, we sought to investigate the oral microbiota structure of children with cleft lip and palate (CLP) and explore the pre-operative oral bacterial composition related to the prognosis of alveolar bone grafting. In total, 28 patients (19 boys, 9 girls) with CLP who were scheduled to undergo alveolar bone grafting for the first time were recruited. According to the clinical examination of operative sites at the third month after the operation, the individuals were divided into a non-inflammation group (n = 15) and an inflammation group (n = 13). In all, 56 unstimulated saliva samples were collected before and after the operation. The v3-v4 hypervariable regions of the 16S rRNA gene were sequenced using an Illumina MiSeq sequencing platform. Based on the beta diversity of the operational taxonomic units (OTUs) in the inflammation and non-inflammation samples, the microbial variation in the oral cavity differed significantly between the two groups before and after the operation (P < 0.05). Analysis of the relative abundances of pre-operative OTUs revealed 26 OTUs with a relative abundance higher than 0.01%, reflecting a significant difference of the relative abundance between groups (P < 0.05). According to a principal component analysis of the pre-operative samples, the inflammation-related OTUs included Tannerella sp., Porphyromonas sp., Gemella sp., Moraxella sp., Prevotella nigrescens, and Prevotella intermedia, most of which were enriched in the inflammation group and showed a significant positive correlation. A cross-validated random forest model based on the 26 different OTUs before the operation was able to fit the post-operative status of grafted sites and yielded a good classification result. The sensitivity and specificity of this classified model were 76.9% and 86.7%, respectively. These findings show that the oral microbiota profile before alveolar bone grafting may be related to the risk of post-operative inflammation at grafted sites.

Prevalence and bacteriology of bacteremia associated with cleft lip and palate surgery.

The aim of the study was to determine the prevalence and bacteriology of bacteremia associated with cleft lip and palate (CLP) surgery. Three venous blood samples were obtained from 90 eligible subjects who presented for CLP surgery: before surgical incision, 1 minute after placement of the last suture, and 15 minutes thereafter. The samples were injected into an Oxoid Signal blood culture and transported to the laboratory for gram-positive/negative and aerobic/anaerobic bacteria analysis. Prevalence of bacteremia associated with cleft surgery was 38.1%. Prevalence rates of bacteremia in cleft lip surgery, cleft palate surgery, and alveoloplasty were 40.9%, 33.3%, and 50%, respectively. There was no significant difference in prevalence rate of positive blood culture in cleft lip surgery, cleft palate surgery, and alveoloplasty (P = 0.69). Positive blood culture was detected most frequently (47%) 1 minute after placement of the last suture. Of the 23 subjects who had positive blood culture at 1 minute, bacteremia persisted in 8 (35%) of them after 15 minutes. The most common bacteria isolated were coagulase-negative staphylococcus, Acinetobacter lwoffii, and coagulase-positive Staphylococcus aureus. Sex and age of the subjects, duration of surgery, blood loss, and type of cleft surgery were not significantly associated with positive blood culture. Bacteremia associated with CLP surgery is polymicrobial and persisted for at least 15 minutes after surgery in 35% of cases. This may reinforce the need for prophylactic antibiotics to protect at-risk patients from developing focal infection of the heart by oral flora.

A novel delivery system of probiotic drop and its effect on dental caries risk factors in cleft lip/palate children.

OBJECTIVE: The aim of the present study was to investigate the effect of the probiotic bacterium Lactobacillus reuteri on the levels of salivary mutans streptococci and lactobacilli in children with cleft lip/palate who used the novel drop containing L. reuteri. MATERIAL AND METHODS: The study group consisted of 19 operated cleft lip/palate children aged 4 to 12 years. The study had a double-blind, randomized crossover design, and the experimental period consisted of four consecutive time periods. During periods 2 and 4, consisting of 25 days each, parents were instructed that their children should consume 5 drops per day (0.15 to 0.20 g) of probiotic or placebo drops produced by the same manufacturer. The probiotic drop, BioGaia Reuteri drops, contained L. reuteri DSM 17938 and L. reuteri ATCC PTA 5289 (≥1 × 10(8) CFU/5 drops). The counts of salivary mutans streptococci and lactobacilli were evaluated using the CRT tests. The data were processed with NCSS 2007 software using chi-square and McNemar tests. RESULTS: There was no statistically significant (p > .05) reduction of salivary mutans streptococci and lactobacilli after 25 days of consumption of both drops. CONCLUSIONS: The novel drop containing L. reuteri may not reduce the levels of salivary mutans streptococci and lactobacilli in cleft lip/palate children.

The value of microbiological screening in cleft lip and palate surgery.

OBJECTIVE: This study was performed to investigate whether nasal and oropharyngeal microbiological swabs taken prior to cleft lip and palate surgery correlated with the oronasal flora at the time of surgery and whether specific culture results affected surgical outcome. METHODS: Prospective audit set in two designated U.K. cleft centers each with a single surgeon. Nasal and oropharyngeal microbiological swabs were taken within 2 weeks prior to surgery and again on the operating table. Adverse outcome measures included postoperative pyrexia, wound dehiscence, or fistula formation. RESULTS: One hundred forty-four cases were recruited over 12 months. Nasal swabs cultured organisms significantly more often than oropharyngeal swabs (p < .0001). No significant difference was detected in the number of cases with a positive microbiology culture preoperatively compared with perioperative sampling (48% and 50%). The specific organisms cultured from preoperative swabs were the same as those cultured at surgery in only half of cases. Preoperative microbiology swabs were poorly predictive of the oronasal flora at surgery. Antibiotic treatment of patients with positive preoperative microbiology did not significantly reduce the incidence of bacterial colonization or significantly alter clinical outcome. CONCLUSION: Preoperative microbiological investigation is not helpful in predicting the nasal and oropharyngeal flora at the time of surgery. Further, culture results did not correlate with postoperative outcome, regardless of whether pre- or perioperative antibiotic therapy was instigated. This evidence suggests that microbiology screening swabs are an unnecessary investigation.

Oral candidal colonization in cleft patients as a function of age, gender, surgery, type of cleft, and oral health.

PURPOSE: To assess the colonization rate of oral Candida species and the influence of age, gender, oral health status, number of surgeries, and type of cleft. PATIENTS AND METHODS: A prospective study of 60 patients with cleft and 60 control subjects was carried out at the Cleft Centre at King Abdullah University Hospital and the Maxillofacial Unit at Jordan University of Science and Technology between October 2007 and June 2008. Oral health was assessed using the Gingival, Plaque, and Decayed, Missing, and Filled (DMFT/dmft) indexes using World Health Organization criteria. A culture swab was obtained from the tongue and buccal and palatal mucosae. Candida albicans and other Candida species were identified using the germ tube test and the automated biochemical test panel VITEK. RESULTS: The colonization rate of Candida in patients with cleft (63.3%) was significantly higher than in healthy control subjects (18.3%). The colonization rate of Candida and the distribution of C albicans varied with age but were not significantly associated with gender in patients with cleft and healthy controls. The candidal colonization rate was highest in patients with cleft who had at least 3 surgeries (78.2%) and in patients with bilateral clefts (77.7%). Patients with cleft had a significantly poorer health status than healthy controls; however, this was not influenced by the type of the cleft or the number of surgeries. CONCLUSION: Patients with cleft had a significantly higher rate of oral candidal colonization compared with control subjects, which varied with age, type of cleft, and the number of surgical interventions. Oral health status was significantly poorer in patients with cleft.