Lyticase Facilitates Mycobiome Resolution Without Disrupting Microbiome Fidelity in Primates

Pierre, Joseph F.; Phillips, Gregory J; Chandra, Lawrance C.; Rendina, Danielle N.; Thomas-Gosain, Neena; Lubach, Gabriele R.; Lyte, Mark; Coe, Christopher L.; Gosain, Ankush

doi:10.1016/j.jss.2021.06.023

Cited by 2 publications

(3 citation statements)

References 15 publications

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“…For mouse samples, we collected the whole left lung and terminal ileum [19]. Microbial DNA was extracted, amplified, and sequenced as described [20, 21]. Negative and positive controls were collected and passed through the DNA extraction and sequencing steps.…”

Section: Methodsmentioning

confidence: 99%

“…We used ZymoBIOMICS whole organism and DNA microbial community standards (Zymo Research) as well as sterile-filtered PBS to create pairs of positive and negative controls appropriate for each step of the sample collection and isolation procedure. Microbial DNA was extracted using the ZymoBIOMICS DNA Miniprep Kit (Zymo Research) with the addition of a lyticase (200 Units per 1 mL sample volume) predigestion with 10-minute bead bashing on a Fisher Vortex Genie 2 (Thermo Fisher Scientific) at RT, then 37 C with gentle agitation for 20 min, as previously validated to facilitate extraction of fungal DNA [16]. Extracted DNA was sequenced using the MiSeq platform (Illumina) at Novogene (human samples) or Argonne National Laboratory (mouse samples) using 16S V3-4 (bacterial) rRNA with primers 515F-806R and the internal transcribed spacer (ITS) 2 (fungal) rRNA gene primers.…”

Section: Methods Detailsmentioning

confidence: 99%

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The fungal intestinal microbiota predict the development of bronchopulmonary dysplasia in very low birthweight newborns

Willis

Silverberg

Martin

et al. 2023

Preprint

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Rationale: Bronchopulmonary dysplasia (BPD) is the most common morbidity affecting very preterm infants. Gut microbial communities contribute to multiple lung diseases, and alterations of the gut microbiome may be a factor in BPD pathogenesis. Objective: To determine if features of the multikingdom gut microbiome predict the development of BPD in very low birthweight newborns. Methods: We performed a prospective, observational cohort study comparing the multikingdom fecal microbiota of 147 very low birthweight infants with BPD or post-prematurity respiratory disease (PPRD) by sequencing bacterial 16S and fungal ITS2 ribosomal RNA. To address the potential causative relationship between gut dysbiosis and BPD, we used fecal microbiota transplant in an antibiotic-pseudohumanized mouse model. Comparisons were made using RNA sequencing, quantitative confocal microscopy, lung morphometry, and oscillometry. Measurements and Main Results: We analyzed 100 fecal microbiome samples collected during the second week of life. Infants who later developed BPD showed an obvious fungal dysbiosis as compared to infants with PPRD (P = 0.0209). Instead of fungal communities dominated by Candida and Saccharomyces, the microbiota of infants who developed BPD were characterized by the abundance of Aureobasidium and a greater diversity of rarer fungi in less interconnected community architectures. On successful colonization, the gut microbiota from infants with BPD augmented lung injury in the offspring of recipient animals. We identified alterations in the murine lung and intestinal microbiomes and transcriptional alterations associated with augmented lung injury. Conclusions: The gut fungal microbiome of infants who will develop BPD is dysbiotic and may contribute to disease pathogenesis.

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Section: Methodsmentioning

confidence: 99%

Section: Methods Detailsmentioning

confidence: 99%

The fungal intestinal microbiota predict the development of bronchopulmonary dysplasia in very low birthweight newborns

Willis

Silverberg

Martin

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Third, previous reports from humans state fungi abundances in the gut are relatively low compared to bacteria, hence possibly environmental contaminants will be most of the amplified and sequenced material (Auchtung et al, 2018;Nilsson et al, 2019). Perhaps, including a lyticase treatment to degrade the fungal cell wall during DNA extraction can increase the amount of material without altering the bacterial community (Pierre et al, 2021).…”

Section: The Unexplored Gut Fungi From Redfronted Lemursmentioning

confidence: 98%

Social relationships: key to gut microbiome composition in wild redfronted lemurs?

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Lyticase Facilitates Mycobiome Resolution Without Disrupting Microbiome Fidelity in Primates

Cited by 2 publications

References 15 publications

The fungal intestinal microbiota predict the development of bronchopulmonary dysplasia in very low birthweight newborns

The fungal intestinal microbiota predict the development of bronchopulmonary dysplasia in very low birthweight newborns

Social relationships: key to gut microbiome composition in wild redfronted lemurs?

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