Ecological competition in the oral mycobiome of Hispanic adults living in Puerto Rico associates with periodontitis.

Background: Fungi are a major component of the human microbiome that only recently received attention. The imbalance of indigenous fungal communities and environmental fungi present in the oral cavity may have a role in oral dysbiosis, which could exacerbate oral inflammatory diseases. Methods: We performed a cross-sectional study and recruited 88 participants aged 21 to 49 from sexually transmitted infection clinics in Puerto Rico. A full-mouth periodontal examination following the NHANES protocol defined periodontal severity (CDC/AAP). ITS2 (fungal) genes were amplified and sequenced for mycobiota characterization of yeast and environmental fungi. Environmental outdoor spore levels were measured daily by the American Academy of Allergy Asthma and Immunology San Juan station and defined by quartiles as spore scores. Results: Our data indicate polymicrobial colonization of yeast and environmental fungi in the oral cavity. Dominant taxa associated with periodontal disease included Saccharomyces cerevisiae, Rigidoporus vinctus, and Aspergillus penicilloides, while Candida albicans were found to be ubiquitous. Fungal aerosols were found to impact the oral cavity biofilm, likely due to competition and neutralization by inhaled outdoor and indoor fungal spores. Conclusion: To our knowledge, this is the first report showcasing the ecological competition of measured outdoor environmental fungi with the human oral mycobiota.

Prenatal exposure to Hurricane Maria is associated with an altered infant nasal microbiome.

Background: Prenatal adverse exposures have been associated with increased risks of development of respiratory diseases in children. The infant nasal microbiome is an important mechanism and indicator. Objective: Our aim was to characterize and compare the nasal microbiome of infants who were in utero and exposed to Hurricane Maria in Puerto Rico during 2017 with that of infants who were conceived at least 5 months after the hurricane as controls. Methods: We recruited 63 vaginally born infants, 29 of whom were in the exposure group and 34 of whom were in the control group. Nasal swab samples were collected and analyzed by using 16S ribosomal RNA gene sequencing at the community and taxon levels, respectively. Results: Infants in the exposure group were more likely to harbor a Staphylococcus-Streptococcus-dominant microbial community in their nose. The richness and diversity of the microbiome was significantly higher in the exposure group than in the control group. In the exposure group, the bacterial genera Rhodocista, Azospirillum, Massilia, Herbaspirillum, Aquabacterium, and Pseudomonas were enriched, whereas Corynebacterium and Ralstonia were depleted. Food insecurity due to Hurricane Maria was associated with an increase in Pseudomonas in the infant nasal microbiome. Conclusion: Infants who were exposed to Hurricane Maria during gestation had an altered nasal microbiome, with a higher prevalence of environmental bacteria. More research is needed to evaluate the long-term impacts of extreme weather events occurring in the prenatal stage on a child's nasal microbiome and respiratory health.

Hurricane María drives increased indoor proliferation of filamentous fungi in San Juan, Puerto Rico: a two-year culture-based approach.

Extensive flooding caused by Hurricane María in Puerto Rico (PR) created favorable conditions for indoor growth of filamentous fungi. These conditions represent a public health concern as contamination by environmental fungi is associated with a higher prevalence of inflammatory respiratory conditions. This work compares culturable fungal spore communities present in homes that sustained water damage after Hurricane María to those present in dry, non-flooded homes. We collected air samples from 50 houses in a neighborhood in San Juan, PR, 12 and 22 months after Hurricane María. Self-reported data was used to classify the homes as flooded, water-damage or dry non-flooded. Fungi abundances, composition and diversity were analyzed by culturing on two media. Our results showed no significant differences in indoor fungal concentrations (CFU/m3) one year after the Hurricane in both culture media studied (MEA and G25N). During the second sampling period fungal levels were 2.7 times higher in previously flooded homes (Median = 758) when compared to dry homes (Median = 283), (p-value < 0.005). Fungal profiles showed enrichment of Aspergillus species inside flooded homes compared to outdoor samples during the first sampling period (FDR-adjusted p-value = 0.05). In contrast, 22 months after the storm, indoor fungal composition consisted primarily of non-sporulated fungi, most likely basidiospores, which are characteristic of the outdoor air in PR. Together, this data highlights that homes that suffered water damage not only have higher indoor proliferation of filamentous fungi, but their indoor fungal populations change over time following the Hurricane. Ultimately, after nearly two years, indoor and outdoor fungal communities converged in this sample of naturally ventilated homes.

Comparison of Atmospheric Fungal Spore Concentrations between Two Main Cities in the Caribbean Basin.

OBJECTIVE: Fungal spores are ubiquitous in the atmosphere worldwide, but their distribution is not homogeneous at different locations. Most studies have compared airborne fungal spores ecology in temperate zones, but less is known about the tropics. METHODS: This study compared, through statistical analysis of archived datasets, the predominant fungal groups, patterns and meteorological variables affecting airborne fungal spore concentrations between two major cities in the Caribbean (Havana and San Juan) during the year 2015. RESULTS: In Havana, the predominant fungal group was Cladosporium while in San Juan were basidiospores. Our data provide evidence of differences and similarities in the monthly distribution of airborne spores in Havana and San Juan, but Cladosporium, ascospores y basidiospores had comparable hourly patterns in both cities and were affected by the same meteorological variables. CONCLUSION: Our study provides additional evidence to help design allergy interventions.

Characterizing environmental asthma triggers and healthcare use patterns in Puerto Rico.

Objective: Asthma carries a high burden of disease for residents of Puerto Rico. We conducted this study to better understand asthma-related healthcare use and to examine potential asthma triggers.Methods: We characterized asthma-related healthcare use in 2013 by demographics, region, and date using outpatient, hospital, and emergency department (ED) insurance claims with a primary diagnostic ICD-9-CM code of 493.XX. We examined environmental asthma triggers, including outdoor allergens (i.e., mold and pollen), particulate pollution, and influenza-like illness. Analyses included descriptive statistics and Poisson time-series regression.Results: During 2013, there were 550,655 medical asthma claims reported to the Puerto Rico Healthcare Utilization database, representing 148 asthma claims/1,000 persons; 71% of asthma claims were outpatient visits, 19% were hospitalizations, and 10% were ED visits. Females (63%), children aged ≤9 years (77% among children), and adults aged ≥45 years (80% among adults) had the majority of asthma claims. Among health regions, Caguas had the highest asthma claim-rate at 142/1,000 persons (overall health region claim-rate = 108). Environmental exposures varied across the year and demonstrated seasonal patterns. Metro health region regression models showed positive associations between increases in mold and particulate matter <10 microns in diameter (PM10) and outpatient asthma claims.Conclusions: This study provides information about patterns of asthma-related healthcare use across Puerto Rico. Increases in mold and PM10 were associated with increases in asthma claims. Targeting educational interventions on exposure awareness and reduction techniques, especially to persons with higher asthma-related healthcare use, can support asthma control activities in public health and clinical settings.

Identification of Immunoglobulin E-Binding Proteins of the Xerophilic Fungus Aspergillus penicillioides Crude Mycelial Mat Extract and Serological Reactivity Assessment in Subjects with Different Allergen Reactivity Profiles.

BACKGROUND: Aspergillus penicillioides is a very common indoor xerophilic fungus and potential causative agent of respiratory conditions. Although people are constantly exposed to A. penicillioides, no proteins with allergenic potential have been described. Therefore, we aim to confirm allergic sensitization to A. penicillioides through reactivity in serological assays and detect immunoglobulin E (IgE)-binding proteins. METHODS: In an indirect ELISA, we compared the serological reactivity to A. penicillioides between subjects with specific IgE (sIgE) (group 1, n = 54) and no sIgE reactivity (group 2, n = 15) against commercial allergens. Correlations and principal component analysis were performed to identify associations between reactivity to commercial allergens and A. penicillioides. IgE-binding proteins in A. penicillioides were visualized using Western blotting (WB) in group 1. The IgE-binding proteins with the highest reactivity were analyzed by mass spectrometry and confirmed by transcript matching. RESULTS: There was no statistical significance (p = 0.1656) between the study groups in serological reactivity. Correlations between reactivity to A. penicillioides, dog epithelia, Aspergillus fumigatus, and Penicillium chrysogenum were observed. WB experiments showed 6 IgE-binding proteins with molecular weights ranging from 45 to 145 kDa. Proteins of 108, 83, and 56 kDa showed higher reactivity. Mass spectrometry analysis of these 3 proteins led to the putative identification of NADP-specific glutamate dehydrogenase and catalase B. This was confirmed with transcriptome analysis. CONCLUSIONS: These results provide evidence of the presence of potential allergenic components in A. penicillioides. Further analysis of the putatively identified proteins should reveal their allergenic potential.

Serological Reactivity and Identification of Immunoglobulin E-Binding Polypeptides of Ganoderma applanatum Crude Spore Cytoplasmic Extract in Puerto Rican Subjects.

BACKGROUND: The allergenic potential of Ganoderma applanatum basidiospores has been demonstrated previously in Puerto Rico. However, basidiomycete allergens are not available for inclusion in allergy diagnostic panels. Therefore, we sought to confirm allergic sensitization to G. applanatum crude spore cytoplasmic extract through reactivity in serological assays and detection of immunoglobulin E (IgE)-binding polypeptides. METHODS: Via an indirect ELISA, serological reactivity was compared between groups of individuals with different allergic profiles. Group 1 (n = 51) consisted of individuals with sIgE to the allergens included in the diagnostic panels; group 2 (n = 14) comprised individuals with no sIgE to the allergens tested; and group 3 (n = 22) included individuals with no allergic history. To visualize IgE-binding polypeptides, group 1 sera were examined via Western blotting (WB). Polypeptide bands with the highest reactivity were analyzed by mass spectrometry (MS) for putative identification. RESULTS: The serological reactivity of group 1 was significantly higher than that of group 3 in an indirect ELISA (p = 0.03). Sixty-five percent of group 1 individuals showed reactivity to polypeptide bands in WB. Bands of 81 and 56 kDa had the highest reactivity proportions among the reactive sera, followed by a 45-kDa band. MS analysis of these 3 polypeptides suggests that they are basidiomycete-derived enzymes with aconitate hydratase, catalase, and enolase functions. CONCLUSIONS: G. applanatum spores have allergenic components recognized by Puerto Rican individuals, which could eventually be considered as markers in cases of fungal allergy and be included in diagnostic allergen panels in Puerto Rico and tropical regions.

Mold populations and dust mite allergen concentrations in house dust samples from across Puerto Rico.

Lifetime childhood asthma prevalence (LCAP) percentages in Puerto Rico Health Regions (HR) are substantially higher in northeastern vs. southwestern HR. Higher average relative humidity in the northeast might promote mold and mite exposures and possibly asthma prevalence. To test this hypothesis, mold contamination, Environmental Relative Moldiness Index (ERMI) values were measured in floor dust (n = 26) and dust mite allergen concentrations in bed dust (n = 14). For this analysis, the eight HR were divided into those with LCAP > 30% (n = 3) and < 30% (n = 5). The average ERMI value was significantly greater (Wilcoxon Rank Sum, p < 0.001) in high than in low LCAP HR (14.5 vs. 9.3). The dust mite antigens Der p 1, Der f 1, and Blo t 5 were detected in 90% of bed samples, but the concentrations were not significantly different in high vs. low LCAP HR. Mold exposures might partially explain the differences in LCAP HR in Puerto Rico.

Skin test reactivity of allergic subjects to basidiomycetes' crude extracts in a tropical environment.

Fungal allergies can be detected by the skin prick test with extracts of the organisms, but not all fungi, including the basidiomycetes, are being examined. We determined the level of sensitization to basidiomycetes in allergic subjects and compared their reactivity to commercial extracts commonly used to detect allergies. Crude spore extracts of the basidiomycetes Ganoderma applanatum, Chlorophyllum molybdites, and Pleurotus ostreatus, which are known to release numerous spores, were examined along with commercial extracts on 33 subjects with asthma, allergic or non-allergic rhinitis. Overall, affected subjects showed the highest reactivity to mites (36%), followed by Ganoderma applanatum (30%), grass (27%) Chlorophyllum molybdites (12%) and Pleurotus ostreatus (12%). Allergic rhinitis patients were most reactive to mites (58%), grass (42%), Ganoderma applanatum (25%), Penicillium spp. (25%), and cat (17%). Those with asthma primarily responded to mites (44%), Ganoderma applanatum (44%), grass (33%), and Pleurotus ostreatus (22%). IgE levels correlated with positive basidiomycetes extracts. This finding, coupled with higher reactivity to basidiospores as compared to mitospores, and the similar sensitivities of patients to G. applanatum and mites, suggest that basidiomycetes are important allergen sources in the tropics.

Sensitization to airborne ascospores, basidiospores, and fungal fragments in allergic rhinitis and asthmatic subjects in San Juan, Puerto Rico.

BACKGROUND: Fungal spores are the predominant biological particulate in the atmosphere of Puerto Rico, yet their potential as allergens has not been studied in subjects with respiratory allergies. The purpose of this study was to determine the level of sensitization of subjects with respiratory allergies to these particles. METHODS: Serum samples were drawn from 33 subjects with asthma, allergic rhinitis, or nonallergic rhinitis and 2 controls with different skin prick test reactivity. An MK-3 sampler was used to collect air samples and the reactivity of the sera to fungal particles was detected with a halogen immunoassay. RESULTS: All subjects reacted to at least 1 fungal particle. Thirty-one subjects reacted to ascospores, 29 to basidiospores, 19 to hyphae/fungal fragments, and 12 to mitospores. The median percentage of haloes in allergic rhinitis subjects was 4.82% while asthma or nonallergic rhinitis subjects had values of 1.09 and 0.39%, respectively. Subjects with skin prick tests positive to 3, 2, 1, or no extract had 5.24, 1.09, 1.61, and, 0.57% of haloed particles, respectively. If skin prick tests were positive to basidiomycetes, pollen, animals, or deuteromycetes, the percentages of haloes were 4.72, 4.15, 3.63, and 3.31%, respectively. Of all haloed particles, 46% were unidentified, 25% ascospores, 20% basidiospores, 7% hyphae/fungal fragments, and 2% mitospores. IgE levels and the number of positive skin prick test extracts correlated with the percentage of haloes. CONCLUSION: In tropical environments, sensitization to airborne basidiomycetes, ascomycetes, and fungal fragments seems to be more prevalent than sensitization to mitospores in subjects with active allergies, suggesting a possible role in exacerbations of respiratory allergies.