Rinse Samples Research Articles

Accumulation of microbial hazards and assessment of food hygiene associated with broiler chicken processing at open air food markets in Maputo, Mozambique

The burden of foodborne disease due to the consumption of animal-sourced foods is substantial in low- and middle-income countries (LMICs). Open air markets, while providing fresh and affordable foods, often have unhygienic practices that may contribute to contamination during the slaughter and processing of chicken meat. This study examines whether and how the common practice of rinse water (stored water used for rinsing broiler carcasses during processing) reuse leads to accumulation of pathogens, with potential cross contamination of chicken meat.To assess the accumulation of Campylobacter jejuni/coli, Salmonella spp., and the indicator of fecal contamination, Escherichia coli, in rinse water used during the slaughtering process at open air food markets in Maputo, Mozambique. We conducted a time-series study at three open air food markets. In a first experiment, we collected paired rinse water (N = 70), water used for chicken processing, and broiler chicken carcass (N = 60) samples from 10 vendors at 75-min intervals starting prior to any processing activity. In a second experiment, we collected 100, 50 mL rinse water samples, immediately before and after processing, from 10 vendors. Chicken processing activity and associated hygiene practices were captured through direct observation. Vendors processed 24 chickens per day, on average. In the first experiment, C. jejuni/coli and E. coli were detected in 30 % and 80 % of rinse water samples, respectively, prior to processing (baseline), and no Salmonella was detected. After the first carcass rinse, C. jejuni/coli and E. coli were detected in 100 % of samples, and Salmonella spp. was detected in 42 % of rinse water samples and 48 % of carcass samples. C. jejuni/coli showed an average 0.1 log10 copies (95 % CI 0.0, 0.2) increase in rinse water and carcass samples every 75 min. In the second experiment, no C. jejuni/coli or Salmonella spp. were detected in baseline rinse water samples, and E. coli were detected in 78 % of baseline rinse water samples. After processing the first carcass, C. jejuni/coli were detected in 100 % of remaining samples, Salmonella spp. were detected in 28 % of pre-final rinse and 36 % of post-final rinse samples, and E. coli were detected in 81 % of pre-final rinse and 100 % of post-final rinse samples. Our results reveal that consumers are at a high risk of purchasing chicken meat contaminated with human enteropathogens. Once contaminated, rinse water stays contaminated throughout the day. Low-cost and feasible interventions implemented at the carcass wash step are needed to reduce microbial hazards on chicken meat before purchase.

Association of Fusobacterium nucleatum Levels by ddPCR in Oral Rinse Samples With Periodontal Disease in Oral Squamous Cell Carcinoma Patients and in Controls.

The role of microbiome, particularly Fusobacterium nucleatum (Fn), in periodontal disease and oral squamous cell carcinoma (OSCC) has been recently explored. This study aimed to evaluate the Fn presence and its levels in oral rinse samples from Brazilian OSCC patients and healthy individuals and its association with sociodemographic, clinical, and oral health features. In this case-control study, 80 participants were included, 31 OSCC patients and 49 individuals without a cancer history. Clinical data were collected, and an oral exam was done on a subset of the cohort. Fn levels were evaluated by droplet digital PCR (ddPCR) in oral rinse samples and were categorized as Fn-high or Fn-low based on the median number of copies per reaction. OSCC patients showed higher levels of Fn (68%, p = 0.03) than controls, and all OSCC cases were diagnosed with periodontal disease (100%, p = 1.0). In the univariate analysis, Fn-high level was more frequently present in OSCC cases compared to controls (p = 0.01). It was also observed that Fn-high level OSCC cases were significantly associated with self-reported non-white ethnicity (71.4%, p = 0.01) and had more infiltrative lesions (57.1%, p = 0.02) than Fn-low OSCC cases. Fn-high levels in oral rinse samples, were significantly more prevalent among OSCC than in controls. In OSCC patients, Fn-high levels were associated with non-white ethnicity and lesions with infiltrative clinical aspects. Among OSCC cases, all had periodontal disease.

Factors affecting Kaposi’s sarcoma-associated herpesvirus transmission in rural Ugandan households, a longitudinal study

BackgroundWe report the impact of HIV infection within a household on oral Kaposi’s sarcoma-associated herpesvirus (KSHV) shedding.MethodsWe enrolled 469 individuals from 90 households. Mouthwash rinse samples collected at three monthly visits were analyzed for KSHV DNA using quantitative polymerase chain reaction (qPCR). Generalized linear mixed effects logistic models were applied to analyze factors associated with KSHV ever shedding, and among shedders, always versus intermittent shedding. Linear mixed effects models were applied to models of KSHV viral loads. Intraclass correlation coefficients (ICCs) were calculated to assess the contribution of household-level factors to variations in shedding probabilities. Hotspot analyses of geospatial feature clusters were calculated using Getis-Ord Gi* statistic and visualized using inverse distance weighted interpolation.ResultsAnalyses included 340 KSHV seropositive individuals, aged 3 + years, with qPCR results from 89 households. Forty households had 1 + persons living with HIV (PLWH), while 49 had none. Among participants, 149(44%) were KSHV ever shedders. Of 140 who shed KSHV at two or more visits, 34(24%) were always shedders. Increasing number of KSHV seropositive household members was significantly associated with ever shedding [Odds ratio(OR) (95% Confidence Interval(95%CI)):1.14(1.03,1.26);p = 0.013]. Among KSHV shedders, a statistically significant age-related trend was identified with 10–19 years being more likely to be always shedders (type III test p = 0.039) and to have higher viral loads (type III test p = 0.027). In addition, higher viral loads were significantly associated with increasing number of household members [coefficient(95%CI):0.06(0.01,0.12);p = 0.042], increasing number of KSHV seropositive members [coefficient(95%CI):0.08(0.01,0.15);p = 0.021], and living in households with 1 + PLWH [coefficient(95%CI):0.51(0.04,0.98);p = 0.033]. Always shedders exhibited higher viral loads than intermittent shedders [coefficient(95%CI):1.62(1.19,2.05);p < 0.001], and viral loads increased with the number of visits where KSHV DNA was detected in saliva (type III test p < 0.001). Household-level factors attributed for 19% of the variability in KSHV shedding (ICC:0.191;p = 0.010). Geospatial analysis indicated overlapping hotspots of households with more KSHV seropositive individuals and KSHV shedders, distinct from areas where PLWH were clustered.DiscussionKSHV oral shedding is influenced by multiple factors at the individual, household, and regional levels. To mitigate ongoing KSHV transmission a comprehensive understanding of factors contributing to oral KSHV reactivation and transmission within households is needed.

Oral Yeasts Carriage in Prediabetic Smokers and Nonsmokers

Habitual smoking and prediabetes are independent risk factors for increased oral yeasts carriage (OYC); however, no studies have compared OYC amongst cigarette smokers and nonsmokers with and without prediabetes. The aim was to fill this research gap. Ninety-two participants were included and categorised into 4 groups: group 1, prediabetic (haemoglobin A1c [HbA1c] levels, 5.7%-6.4%) cigarette smokers; group 2, cigarette smokers without prediabetes; group 3, prediabetic nonsmokers; and group 4, nonsmokers without prediabetes. Patient demographics and HbA1c levels were recorded. Data on duration of smoking habit (pack-years) and family history of smoking were collected. Information on daily toothbrushing and flossing and most recent visit to a dentist/dental hygienist was gathered. Clinical and radiographic periodontal examination was performed and unstimulated whole salivary flow rate (UWSFR) was determined. OYC was assessed using the oral rinse sampling method. Power analysis was done, and group comparisons were performed. Logistic regression analysis was performed and P values <5% reflected statistical significance. Respectively, 23, 24, 22, and 23 individuals with comparable mean ages were included in groups 1, 2, 3, and 4. In groups 1 and 2, participants had a smoking history of (mean ± SD) 24.7 ± 3.2 and 10.6 ± 2.5 pack-years. Plaque index, clinical attachment loss, and probing depth were higher in groups 1 (P < .05), 2 (P < .05), and 3 (P < .05) than in group 4. Number of missing teeth was significantly higher in group 1 compared with groups 2 (P < .05), 3 (P < .05), and 4 (P < .05). There was no difference in UWSFR amongst the groups. OYC was greater in group 1 than in groups 2 (P < .05), 3 (P < .05), and 4 (P < .05). OYC was greater in groups 2 (P < .05) and 3 (P < .05) than in group 4. In prediabetic cigarette smokers, OYC appears to be influenced by hyperglycaemia, whilst in nondiabetic smokers, the severity of periodontal inflammation appears to be the determining factor in OYC.

Isolation and Identification Of Candida Albicans In Smokers With Epithelial Dysplasia And Squamous Cell Carcinoma: A Histopathological Correlation

Background:Greater adaptability of Candida to the host niche makes it the most common opportunistic pathogen. Its role as a commensal or pathogen in development of premalignancies and its progression to malignancies is of considerable debate. Present study was undertaken to determine the role of Candida albicans(CA)in progressionof oral cancer by isolation and identification by mycological methods in smokers without oral lesions and histopathologically diagnosed cases of epithelial dysplasia (ED) and oral squamous cell carcinoma (OSCC). Methodology:The study comprised of 120 cases that included study group of 40 smokers with oral lesions, 40 smokers without oral lesions and 40 non-smokers as control group. Clinical examination was followed by oral rinse sample collection and biopsy for histopathological confirmation. Sedimented oral rinse was inoculated on Sabouraud’s dextrose agar (SDA). Isolated Candida colony after identification with Gram stain was subjected to germ tube and chlamydospore formation test for confirmation of CA. The data was statistically analysed using chi square test. Results:Colonisation of CA was 52.5% in the study group and 5% in the control group, was found to be statistically significant (p&lt;0.001).Association of CA in smokers with varying histopathologic grades of oral lesions showed no evidence of a correlation. However, positive correlation was found in OSCC (n=19, 95%) and showed statistical significance (p&lt;0.001). Conclusion:The present study showed an increase in colonization of CA in malignant lesions suggesting that CA in conjunction with tobacco usage may play a role in oral carcinogenesis.

Is Short-Read 16S rRNA Sequencing of Oral Microbiome Sampling a Suitable Diagnostic Tool for Head and Neck Cancer?

The oral microbiome, studied by sampling the saliva or by oral rinse, has been long thought to have diagnostic capacity for head and neck cancers (HNC). However, previous reports on the HNC oral microbiome provide inconsistent results. The aim of this study is to consolidate these datasets and determine the oral microbial composition between HNC patients to healthy and premalignant individuals. We analyzed 16 published head and neck cancer (HNC) short-read 16S rRNA sequencing datasets, specifically targeting the V3V4, V4 and V4V5 regions. These datasets included saliva and oral rinse samples from donors with HNC, as well as from healthy and premalignant donors. Differences in diversities and microbial abundance were determined. HNC saliva displayed lower alpha diversity than healthy donors. In contrast, the opposite trend was observed for oral rinse samples. Beta diversity scores were largely similar across different patient types. Similar oral phyla were detected for all samples, but proportions were largely dependent on sample type (i.e., saliva or oral rinse) and primer set utilized for 16S rRNA sequencing. Neisseria, Leptotrichia and Megasphaera were elevated in healthy saliva, while Mycoplasma was elevated in HNC saliva. Oral rinse and saliva displayed similar enrichment for Fusobacterium, while Veillonella, Alloprevotella, and Campylobacter showed conflicting results. The sparse partial least squares discriminant analysis model performed effectively in discriminating HNC from healthy or premalignant patients using V3V4 saliva (AUC = 0.888) and V3V4 oral rinse (AUC = 0.928), while poor discriminative capacity was observed for V4 saliva (AUC = 0.688). In conclusion, our meta-analysis highlighted the limitations of 16S rRNA sequencing, particularly due to variations across study batches, primer sets (i.e., V3V4, V4), and sample types. Hence, caution should be exercised when interpreting 16S rRNA sequencing results across studies, especially when different primer sets and sample types are used.

Human Papilloma Virus (HPV) Detection in Oral Rinse vs. Oral Sponge: A Preliminary Accuracy Report in Oral Cancer Patients

Background/Objectives: Human Papillomavirus (HPV) is a significant etiological factor in the development of oropharyngeal carcinogenesis. The detection of HPV in oral squamous cell carcinoma (OSCC) could be also crucial for diagnosis, prognosis, and treatment planning. This study compares the efficacy and accuracy of two non-invasive sampling methods, oral rinse, and oral sponge, in detecting HPV DNA in patients with OSCC. Methods: Twenty-six patients with histologically confirmed OSCCs were recruited (M/F = 15/11; mean age 68.6). From each patient, two self-collected oral specimens, in the form of an oral rinse and a salivary sponge (i.e., LolliSponge), were collected, and subsequently processed, utilizing INNO-LiPA HPV Genotyping Extra II for HPV DNA detection; Results: Oral sponge detection showed high specificity (100%), sensitivity (85.7%), and accuracy (96.2%) compared to the oral rinse sampling, also demonstrating an area AUC for its diagnostic performance significantly greater than 0.5 (0.93 vs. 0.5, p &lt; 0.0001). Conclusions: This study supports that oral sponge sampling offers valuable non-invasive alternatives for HPV detection in patients with OSCC, with the potentiality to facilitate saliva sampling in patients that may exhibit functional deficit due to OSCC. Further research is recommended to validate these findings in larger cohorts and to explore the integration of these methods into routine clinical practice for the management of HPV-related OSCC.

Evaluation of Methods for Identifying Poultry Wing Rinses With Salmonella Concentrations Greater Than or Equal to 10 CFU/mL

In the United States, the Proposed Regulatory Framework to Reduce Salmonella Illnesses Attributable to Poultry published by the Food Safety and Inspection Service (FSIS) has highlighted the need for simple, rapid methods that identify poultry wing rinse samples harboring Salmonella concentrations ≥10 CFU/mL. One of eight cold-stressed and nutrient-starved Salmonella strains was inoculated into post-chill two-joint poultry wing rinses (48 turkey and 72 chicken) at levels from 0.22 to 3.79 log CFU/mL, and then measured by 3-tube Most Probable Number (MPN), BioMerieux GENE-UP QUANT, Hygiena BAX SalQuant, and novel threshold methods. The MPN lower limit of quantification (LLQ) for Salmonella was −0.96 log CFU/mL. MPN overestimated the inoculated Salmonella level by 0.05 ± 0.35 log CFU/mL. The GENE-UP QUANT Salmonella method (LLQ = 1.00 log CFU/mL) underestimated the inoculated Salmonella level by 0.05 ± 0.51 log CFU/mL. The BAX SalQuant method (LLQ = 0.00 log CFU/mL) underestimated the inoculated Salmonella level by 1.21 ± 0.78 log CFU/mL. Threshold test methods with Poisson probabilities of 0.95 (PiLOT-95), 0.86 (PiLOT-86), 0.63 (PiLOT-63), and 0.50 (PiLOT-50) were developed to identify poultry wing rinses harboring Salmonella levels ≥10 CFU. MPN was 93.1%, accurate for determining if Salmonella levels in poultry wing rinses were ≥10 CFU/mL, but MPN costs and time requirements can be prohibitive for most laboratories. GENE-UP quantification was 86.1% accurate, but the GENE-UP method requires equipment and technical expertise that some food safety laboratories may not possess. BAX quantification had the lowest accuracy; 58.4%. PiLOT threshold test accuracies ranged from 83.2% for PiLOT-50 to 93.1% for PiLOT-86. The PiLOT threshold tests are simple and can be adapted to identify many environmental or food samples containing Salmonella exceeding any user-defined concentration threshold.

Factors affecting Kaposi's sarcoma-associated herpesvirus transmission in rural Ugandan households, a longitudinal study.

We report the impact of HIV infection within a household on oral Kaposi's sarcoma-associated herpesvirus (KSHV) shedding. We enrolled 469 individuals from 90 households. Mouthwash rinse samples collected at three monthly visits, were analyzed for KSHV DNA using quantitative polymerase chain reaction (qPCR). Generalized linear mixed effects logistic models were applied to analyze factors associated with KSHV ever shedding, and among shedders, always versus intermittent shedding. Linear mixed effects models were applied to models of KSHV viral loads. Intraclass correlation coefficients (ICCs) were calculated to assess the contribution of household-level factors to variations in shedding probabilities. Hotspot analyses of geospatial feature clusters were calculated using Getis-Ord Gi* statistic and visualized using inverse distance weighted interpolation. Analyses included 340 KSHV seropositive individuals, aged 3 + years, with qPCR results from 89 households. Forty households had 1 + persons living with HIV (PLWH), while 49 had none. Among participants, 149(44%) were KSHV ever shedders. Of 140 who shed KSHV at two or more visits, 34(24%) were always shedders. Increasing number of KSHV seropositive household members was significantly associated with ever shedding [Odds ratio(OR) (95% Confidence Interval(95%CI)):1.14(1.03,1.26);p = 0.013]. Among KSHV shedders, a statistically significant age-related trend was identified with 10-19 years being more likely to be always shedders (type III test p = 0.039) and to have higher viral loads (type III test p = 0.027). In addition, higher viral loads were significantly associated with increasing number of household members [coefficient(95%CI):0.06(0.01,0.12);p = 0.042], increasing number of KSHV seropositive members [coefficient(95%CI):0.08(0.01,0.15);p = 0.021], and living in households with 1 + PLWH [coefficient(95%CI):0.51(0.04,0.98);p = 0.033]. Always shedders exhibited higher viral loads than intermittent shedders [coefficient(95%CI):1.62(1.19,2.05);p < 0.001], and viral loads increased with the number of visits where KSHV DNA was detected in saliva (type III test p < 0.001). Household-level factors attributed for 19% of the variability in KSHV shedding (ICC:0.191;p = 0.010). Geospatial analysis indicated overlapping hotspots of households with more KSHV seropositive individuals and KSHV shedders, distinct from areas where PLWH were clustered. KSHV oral shedding is influenced by multiple factors at the individual, household, and regional levels. To mitigate ongoing KSHV transmission a comprehensive understanding of factors contributing to oral KSHV reactivation and transmission within households is needed.

Abstract P382: The protective role of elevated oral microbial levels in adult hypertension: NHANES 2009-2012

Background and Objective: Accumulating evidence has linked periodontal disease with hypertension, yet the relationship between the diversity of oral microbiota and blood pressure remains unclear. Our objective was to investigate the association between characteristics of oral microbiome diversity and hypertension in the general population. Methods: The data were extracted from the 2009-2012 National Health and Nutrition Examination Survey (NHANES), where we included participants aged 20 years or older who had indicators of hypertension. Participants with conditions such as pregnancy, periodontal disease, cardiovascular diseases (CVD), chronic obstructive pulmonary disease (COPD), diabetes, and cancer were excluded from the analysis. Oral microbiome α-diversity in adults was analyzed through oral rinse samples, encompassing measures such as observed amplicon sequence variants (ASVs), Faith’s Phylogenetic Diversity, the Shannon-Wiener Index, and the Simpson Index. Differences in oral microbiome α-diversity among various hypertension groups were described using weighted mean values. The association between oral microbiome α-diversity and hypertension was analyzed using multivariable logistic regression models. Results: Between 2009 and 2012, among 3,228 participants (weighted mean [SE] age, 36.54 [0.64] years; 1,408 males [weighted 43.62%]), there were 601 cases of hypertension (increasing to 771 when adopting the new diagnostic criteria for hypertension). Significant differences were observed in the levels of oral microbiota α-diversity among different hypertension groups. Multivariate logistic regression analyses revealed an independent association between levels of oral microbiome α-diversity and the risk of hypertension, after controlling for confounding factors. For each one-standard deviation (SD) increase in ASVs and Faith's Phylogenetic Diversity, the adjusted odds ratios (aORs) were 0.85 (95% confidence interval [CI]: 0.75-0.95) and 0.85 (95% CI: 0.75-0.95), respectively. This association persisted across varying diagnostic criteria for hypertension. Conclusions: Our findings indicate diverse oral microbiomes among hypertension groups, where greater α-diversity inversely correlates with hypertension risk, independent of diagnostic criteria. This implicates oral microbiota dysbiosis, potentially from environmental factors, as an additional hypertension risk factor, emphasizing the oral microbiome's potential in hypertension development.

Oral specimens as a tool for accurate metagenomic analysis: A pilot study

ObjectivesAcute oral mucosal damage, as well as other inflammatory processes seem to be related to dysbiosis of the oral microbiome. The need to study changes in the oral microbiome led us to hypothesize what type of sample would provide the most representative picture of the entire human oral microbiome. Materials and methodsAn observational, and cross-sectional study was carried out. Six healthy adult participants provided 3 different sample types each, that included saliva, oral rinse and mucosal biopsy tissue. We performed 16S rRNA sequencing of the V3-V4 region of the 18 samples using Illumina MiSeq technology. ResultsParticipants were 27 ± 6,3 years old. Bacterial alpha diversity was higher in oral rinse samples compared to whole unstimulated saliva and oral mucosa tissue (p = 0,005). However, saliva specimens showed a 56 % relative abundance of identified species followed by a 30 % in oral rinse and only 1 % in tissue samples. ConclusionsThis study found differences on oral microbiome composition for each type of sample. Oral rinse should be chosen when higher alpha diversity is needed, whereas whole unstimulated saliva should be more appropriate for larger amount of bacterial DNA. Clinical relevanceThe results obtained demonstrate the importance of a correct choice of the optimal type of oral sample for microbiome studies due to the differences found in its composition.

Axitinib plus avelumab for recurrent/metastatic adenoid cystic carcinoma (R/M ACC): Biomarker analysis of the phase II trial.

6104 Background: In the phase II axitinib (VEGFR inhibitor) plus avelumab (PD-L1 inhibitor) trial in R/M ACC, response rate was 18% with a median progression-free survival (PFS) of 7.3 months, with subsequent inclusion of the combination in NCCN guidelines as a possible treatment for ACC (category 2B). We sought to identify potential biomarkers predictive of axitinib plus avelumab benefit in R/M ACC. Methods: The phase II trial included 28 R/M ACC patients (pts). Pre-treatment tumors were analyzed by whole exome sequencing (n=24), using Twist Human Core Exome V2 kit, and transcriptome profiling (n=26), using HTG Transcriptome Panel (19,616 probes). For 17 pts, imaging mass cytometry (IMC) was performed with 35 metal-tagged markers and analyzed with Visiopharm software. Microbiome composition was assessed in tumor (n=20), oral rinse (n=20) and fecal (n=19) samples via 16s rRNA gene sequencing. All results were assessed for association with PFS on therapy with a Cox proportional hazards model, maintaining ACC subtype (ACC-I vs. ACC-II) as a covariate. Results: Only 1 (6%) of 17 assessed tumors were PD-L1 positive. By WES, tumor mutational burden (TMB) was overall low (median 1.4 mut/Mb, range 0.7 – 18.7), but higher TMB was associated with worse PFS. Four mutational signatures significantly correlated with worse PFS, including SBS86, associated with previous chemotherapy exposure. Immune deconvolution of RNA-seq data revealed that a higher neutrophil-to-lymphocyte ratio and more T follicular helper cells associated with worse PFS, while presence of cytotoxic cells, T effector memory cells, and mast cells associated with longer PFS. Notably, a 167-gene predictive signature was developed and validated in the phase II ipilimumab plus nivolumab salivary gland cancer trial. This 167-gene signature was not prognostic in ACC pts who did not receive immunotherapy. IMC single-cell immune mapping revealed that pts with longer PFS had increased presence of CD8 T cells, CD73+ macrophages in tumor, a higher stromal CD8 T cell:Regulatory T cell ratio, increased stromal SIGLEC15 positive cells, and a higher tumor-to-stroma ratio of fibroblasts. Conversely, Ki67-positive T cells, cancer stem cells, and M2 macrophages were associated with worse PFS. Through microbiome analysis, 13 bacterial genera in tumor samples, 25 in stool, and 8 in oral rinse were significantly associated with therapy outcomes. Of note, in stool microbiome, presence of Akkermansia spp. and Bifidobacterium spp. were significantly correlated with improved PFS, with the latter association also seen in oral rinse. Conclusions: Correlative analysis of the phase II axitinib plus avelumab trial revealed potential biomarkers predictive of combination clinical benefit in ACC, including a gene-expression signature. These findings may guide patient stratification for combinatorial therapy.

Association of Periodontitis and Various Genotypes of Human Papillomavirus in Oral Rinse Specimens.

Objectives To evaluate the relationship between the presence of human papillomavirus (HPV) genotypes in oral rinse samples and periodontitis. Materials and methods This cross-sectional study utilized data from the National Health and Nutrition Examination Survey (NHANES) conducted in 2013-2014. The primary outcome was the periodontal status, categorized as either no periodontitis or mild periodontitis (combined) versus moderate to severe periodontitis (combined). The primary variable of interest was the presence of different HPV types in oral rinse specimens. Several confounders were selected based on previous evidence that demonstrated a potential association between HPV infectivity and periodontal disease. Results The final sample included 3103 participants. HPV genotypes 6, 35, 39, 55, 59, 71, 72, and 73 showed a statistically significant association with at least one of the periodontal statuses (p-value < 0.05). The presence of any HPV genotype was highly significantly associated with periodontal status, with a p-value of <0.001. Multivariable logistic regression analysis revealed statistically significant associations of HPV 6 and HPV 16 with periodontitis, adjusted for age, gender, diabetes, smoking, race, federal poverty level, last dental visit, and education level. Conclusion Our study highlights a potential link between periodontitis and specific HPV genotypes, such as HPV 6 and HPV 16, in oral rinse specimens. This suggests a complex interplay between periodontal disease and oral HPV infections, underscoring the need for further research to address public health concerns and inform preventive and treatment strategies. Clinical relevance Identifying a link between periodontitis and specific HPV genotypes, such as HPV 6 and HPV 16, in oral rinse samples could prompt early screening and tailored treatment approaches. This underscores the importance of oral health promotion and targeted interventions to address both conditions and improve overall patient outcomes.

Epigenome-wide association study of lung cancer among never smokers in two prospective cohorts in Shanghai, China

BackgroundThe aetiology of lung cancer among individuals who never smoked remains elusive, despite 15% of lung cancer cases in men and 53% in women worldwide being unrelated to smoking. Epigenetic...

Applying the modified UV-activated TOP assay to complex matrices impacted by aqueous film-forming foams

Per- and polyfluoroalkyl substances (PFAS) are a large chemical family, and numerous chemical species can co-exist in environmental samples, especially those impacted by aqueous film-forming foams (AFFFs). Given the limited availability of chemical standards, capturing the total amount of PFAS is challenging. Thus, the total oxidizable precursor (TOP) assay has been developed to estimate the total amount of PFAS via the oxidative conversion of precursors into perfluorocarboxylic acids (PFCAs). This study aims to enhance the robustness of the TOP assay by replacing heat activation with UV activation. We evaluated the molar yields of known precursors in water in the presence of varying levels of Suwannee River natural organic matter (SRNOM) and in two soils. The impact of UV activation was also evaluated in two soils spiked with three well-characterized AFFFs, six AFFF-impacted field soils, and nine rinse samples of AFFF-impacted stainless-steel pipe. In the presence of 100 mg/L SNROM, 6:2 fluorotelomer sulfonate (FTS), 8:2 FTS, and N-ethyl perfluorooctane sulfonamidoacetic acid (N-EtFOSAA) in deionized water had good molar recovery as PFCAs (average of 102 ± 9.8 %); at 500 mg/L SNROM, the recovery significantly dropped to an average of 51 ± 19 %. In two soils (with 4 % and 8.8 % organic matter) with individual precursor spikes, the average molar recovery was 101 ± 9.4 %, except N-EtFOSAA, which had a reduced recovery in the soil with 8.8 % organic matter (OM). UV-activated assays outperformed heat-activated ones, especially in AFFF-impacted soils and pipe extract samples, with an average of 1.4–1.5× higher PFCA recovery. In almost all test samples, UV activation resulted in a notable shift towards longer PFCA chain lengths, particularly for samples with high OM content. The study confirmed the advantages of UV activation, including a significantly shortened exposure time (1 h vs. 6 h) and reduced matrix effects from OM due to the dual functions of UV in activating persulfate and photodegrading OM.

Targeted Next-Generation Sequencing Assay for Direct Detection and Serotyping of Salmonella from Enrichment

In this study, an automated, targeted next-generation sequencing (tNGS) assay to detect and serotype Salmonella from sample enrichments was evaluated. The assay generates millions of reads to detect multiple Salmonella-specific genes and serotype-specific alleles, detecting all Salmonella spp. tested to date, and serotyping 62 common Salmonella serotypes. Accuracy was tested on 291 pure reference cultures (251 Salmonella, 40 non-Salmonella), 21 artificially contaminated poultry carcass rinse samples, and 363 naturally contaminated poultry environmental samples. Among the 291 pure reference cultures, the automated tNGS assay resulted in 100% detection accuracy, 100% serotyping accuracy for the claimed serotypes, and 0% false positives. The limit of detection was estimated at 5 × 104 CFU/mL by testing enumerated cultures of strains representative of six serotypes. In cocontamination studies with mixtures of two serotypes (Enteritidis, Typhimurium, Kentucky, Infantis, and Newport) at a 1:1 ratio, tNGS detected both serotypes with 100% accuracy. The assay demonstrated 100% accuracy in artificially contaminated poultry carcass rinse sample enrichments. Targeted NGS was highly effective in detecting Salmonella in samples collected from poultry production facilities. Results demonstrated that tNGS could detect Salmonella and provide accurate serotyping information consistent with conventional serology. These findings highlight the reliable and efficient performance of a fully automated tNGS Salmonella assay in detecting and identifying Salmonella strains in complex matrices, reducing the time to results from 4 to 5 days required by the traditional isolation and serotyping to 10–12 h for tNGS after primary enrichment.

Decreased GCF DEL-1 and increased GCF neutrophils with increasing probing pocket depth.

Developmental endothelial locus-1 (DEL-1) plays a role in regulating neutrophil migration within the periodontium. The objective of this study was to evaluate the levels of DEL-1 in saliva and gingival crevicular fluid (GCF), as well as the number of neutrophils in patients with periodontitis. Forty systemically healthy, non-smoking periodontitis patients participated in this study. Clinical periodontal parameters, including the plaque index, probing pocket depth (PPD), clinical attachment level, bleeding on probing, modified sulcular bleeding index, and marginal bone level, were measured. Levels of DEL-1, interleukin (IL)-1β, IL-6, and IL-8 in unstimulated saliva samples, as well as DEL-1 in the GCF of 3 teeth from each participant, were assessed. Neutrophil counts in oral rinse and GCF samples were recorded. Spearman correlation coefficients were used to examine the correlation between protein levels, clinical parameters, and neutrophil quantities. Participants were divided into 2 age groups (those under 50 years and those 50 years or older) in order to investigate potential age-related differences. DEL-1 levels in the GCF showed a negative relationship with PPD (sum). Neutrophils in oral rinse samples were positively correlated with PPD, IL-8, and IL-1β levels. Neutrophils in GCF exhibited a positive correlation with PPD (sum). Salivary DEL-1 levels showed correlations with IL-8 and IL-1β, but not with the clinical parameters of periodontitis. The negative relationship observed between PPD and GCF DEL-1 levels is consistent with the proposed protective role of DEL-1.

Dynamic changes of Th1/Th2/Th17 cytokines and hBD-2/3 in erosive oral lichen planus patients saliva before and after prednisone acetate treatment

ObjectiveThis study aimed to investigate the expression of T helper 1 (Th1)/Th2/Th17- related cytokines and human beta defensins 2 and 3 (hBD-2 and -3) in the saliva of patients with erosive oral lichen planus (EOLP) and to explore their role in the pathogenesis of EOLP and the effects of glucocorticoids on EOLP. MethodsA total of 30 patients with EOLP and 20 age- and sex-matched healthy individuals were included in this study. The patients were treated with prednisone at a dose of 0.4 mg/(kg·d) for 1 week and examined before and after treatment. Unstimulated whole saliva samples were collected to determine the levels of cytokines (interleukin 1 beta [IL-1β], tumour necrosis factor alpha [TNF]-α, interferon gamma [IFN-γ], IL-4, IL-6, IL-10 and IL-17) by cytometric bead array and those of hBD-2 and -3 b y enzyme-linked immunosorbent assay. In addition, oral rinse samples were collected to detect Candida load. ResultsThe levels of salivary IL-1β, IL-6, hBD-2 and hBD-3 were higher and the IFN-γ/IL-4 and IL-1β/IL-6 ratios were lower in patients with EOLP than in healthy individuals. In patients with EOLP, hBD-2 levels were positively correlated with IFN-γ levels and negatively correlated with IL-17 levels, whereas hBD-3 levels were negatively correlated with IL-17 and IL-10 levels. In addition, the prevalence of EOLP was positively correlated with IL-6 levels and negatively correlated with the IFN-γ/IL-4 ratio. The levels of IL-1β, TNF-α, IFN-γ, IL-6, hBD-2 and hBD-3 and the IFN-γ/IL-4 ratio decreased after treatment with prednisone for 1 week. The levels of IL-6, hBD-2 and hBD-3 were significantly higher in EOLP patients than in healthy individuals; while TNF-α levels and the IFN-γ/IL-4 ratio were significantly lower in EOLP patients than in healthy individuals. Furthermore, the oral counts of Candida spp. (colony forming unit [CFU]) were negatively correlated with TNF-α levels. Numerical Rating Scale(NRS) and Sign scores decreased in EOLP patients after treatment. Approximately 80 % of patients were effectively treated. Salivary TNF-α levels were significantly higher in the treatment-ineffective group than in the treatment-effective group before treatment with prednisone, and differences in salivary IL-6 levels before and after treatment were significantly higher in the treatment-effective group than in the treatment-ineffective group. ConclusionsHigh expression of IL-1β, IL-6, hBD-2 and Th1/Th2 imbalance in saliva may be associated with the pathogenesis of EOLP. IFN-γ/IL-4 balance may serve as a protective factor for EOLP. Glucocorticoids significantly alleviate the symptoms of EOLP and inhibit the expression of Th1/Th2 cytokines.

Oral Inflammation and Human Papilloma Virus Association among Hispanics.

Identifying factors related to persistent human papillomavirus (HPV) infection is essential to reduce the incidence of HPV-related cancers. To evaluate whether gingival/periodontal inflammation is associated with oral HPV infection. This cross-sectional study (n = 740) uses data from the follow-up visit of the San Juan Overweight Adults Longitudinal Study, which recruited overweight/obese adults aged 40-65 from Puerto Rico. Participants completed a dental examination and two interviews (face-to-face/ACASI) and provided oral rinse samples for HPV detection. Oral inflammation was assessed using two definitions: (1) the number of sites with bleeding on probing (BOP), and (2) the number of teeth with probing pocket depths (PPD) ≥ 4 mm and BOP. Multivariate logistic regression was used to assess the association between oral inflammation and oral HPV. Nearly three-quarters (72%) of participants were female, and 68% had 50 years or older. Participants with HPV had a higher mean number of sites with BOP (15.5 vs. 10.1) and teeth with PPD ≥ 4 mm and BOP (8.5 vs. 3.2) than participants without HPV (p < 0.05). After adjusting for sex, age, income, and the number of oral sex partners, the odds of having an oral HPV infection increased by 3% (95% confidence interval (CI): 1.00-1.06) for any additional sites with BOP and 5% (95% CI: 1.02-1.09) for any other teeth with PPD ≥ 4 mm and BOP. We found that oral inflammation was associated with oral HPV infection among adults in Puerto Rico. Future studies need to further investigate the underlying mechanisms.

Abstract C135: The oral microbiome and short chain fatty acids as contributors to oral HPV disparities in Puerto Rican people with HIV

Abstract Human papillomavirus (HPV) is the most common malignancy worldwide, accounting for more than 70% of oropharyngeal cancers. People living with HIV (PWH) are disproportionately affected by HPV infection and are at higher risk of HPV-associated oral complications, dysplasia, and cancer, even with suppressive antiretroviral therapy (ART), suggesting that ART may not fully recover oral HPV-specific immunity. Particularly, Puerto Rico (PR) has a significant burden and disparity for both HIV and HPV infections and related cancers. Dysbiosis of the oral microbiome occurs in PWH, characterized by a decrease in diversity, and a change in the abundance of microbial products important for human health and disease. The relationship of the oral microbiome with HPV infection is poorly understood, particularly in PWH. In this study, we measured oral microbial diversity and products, such as short chain fatty acids (SCFA), in saliva and evaluated them in the context of HPV infection in PWH. We evaluated 48 sexually active PWH (17 HPV+ and 31 HPV-) from a community clinic in San Juan, Puerto Rico. Oral rinse samples were collected and analyzed for HPV infection and genotyped using the DNA ELISA kit HPV SPF10 and RHA kit HPV SPF10-LiPA25 which detects 40 HPV subtypes. We characterized oral prokaryotic communities by sequencing 16S rDNA and quantified the levels of SCFAs (acetate, propionate, iso-butyrate, butyrate, iso-valerate, valerate, and hexanoate) in saliva using GC-MS. Statistical analyses of bacterial diversity and levels of SCFA were assessed in relation to HPV status using R-statistical software. PWH had a median CD4 count of 738 cells/µL [IQR: 649-887]. PWH with HPV had a 73% (11/15) of high-risk HPV genotypes, with HPV-18 being the most prevalent subtype (6/17). While there was no significant difference in alpha diversity (Shannon: p=0.23 and Observed features: p=0.28), bacterial communities were clustered according to HPV status using a Bray-Curtis dissimilarity index. Furthermore, PWH with HPV showed higher levels of butyrate (p=0.07) and significantly lower levels of hexanoate (p=0.05) in saliva. Particularly, PWH with the HPV-18 subtype had lower levels of hexanoate than other HPV subtypes and HPV- PWH (p=0.03). Overall, these findings suggest that the oral microbiome and SCFA are associated with HPV infection and the presence of specific HPV subtypes. While a difference in bacterial diversity might not be driving HPV infection, different bacterial groupings might affect the abundance of specific SCFA, such as butyrate, which can contribute to local inflammation and periodontal tissue damage, providing an ideal environment for HPV infection and persistence. Understanding how the oral microbiota could facilitate HPV infection and other comorbidities in PWH is crucial for the development of cancer prevention and reduce the burden of HPV-associated malignancies in PWH. Furthermore, HPV vaccine-oriented and oral health educational interventions should target this high-risk group. Citation Format: Yabdiel A. Ramos Valerio, Eduardo S. Perez Delgado, Maria M. Sanchez, Ramon F. Gonzalez Garcia, Ana P. Ortiz Martinez, Magaly Martinez Ferrer, Josue Perez Santiago. The oral microbiome and short chain fatty acids as contributors to oral HPV disparities in Puerto Rican people with HIV [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C135.