Quantification of Bacterial Loads in Caenorhabditis elegans

Walker, Alyssa C; Bhargava, Rohan; Vaziriyan-Sani, Alfonso S.; Brust, Amanda S; Czyż, Daniel M.

doi:10.21769/bioprotoc.4291

Cited by 5 publications

(11 citation statements)

References 7 publications

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“…The 1.5 mL tubes were shaken twice for 90 sec before serial dilution of the lysates and plating onto standard NGM plates. The CFUs/animal values were calculated as described [92]. Data were plotted using Prism 9 and the statistical significance between food sources was determined by one-way ANOVA followed by Sidak’s multiple comparisons test.…”

Section: Methodsmentioning

confidence: 99%

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Kombucha Tea-associated Microbes Remodel Host Metabolic Pathways to Suppress Lipid Accumulation

DuMez-Kornegay,

Morris,

Dowen

2023

Preprint

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The popularity of the ancient, probiotic-rich beverage Kombucha Tea (KT) has surged in part due to its purported health benefits, which include protection against metabolic diseases; however, these claims have not been rigorously tested and the mechanisms underlying host response to the probiotics in KT are unknown. Here, we establish a reproducible method to maintainC. eleganson a diet exclusively consisting of Kombucha Tea-associated microbes (KTM), which mirrors the microbial community found in the fermenting culture. KT microbes robustly colonize the gut of KTM-fed animals and confer normal development and fecundity. Intriguingly, animals consuming KTMs display a marked reduction in total lipid stores and lipid droplet size. We find that the reduced fat accumulation phenotype is not due to impaired nutrient absorption, but rather it is sustained by a programed metabolic response in the intestine of the host. KTM consumption triggers widespread transcriptional changes within core lipid metabolism pathways, including upregulation of a suite of lysosomal lipase genes that are induced during lipophagy. The elevated lysosomal lipase activity, coupled with a decrease in lipid droplet biogenesis, is partially required for the reduction in host lipid content. We propose that KTM consumption stimulates a fasting- like response in theC. elegansintestine by rewiring transcriptional programs to promote lipid utilization. Our results provide mechanistic insight into how the probiotics in Kombucha Tea reshape host metabolism and how this popular beverage may impact human metabolism.

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Section: Methodsmentioning

confidence: 99%

“…Measurement of the bacterial loads in C. elegans animals after consumption of each diet was performed as previously described [92]. Briefly, ~150 animals were grown from synchronized L1s on each diet to adulthood and ~30 animals were picked to an empty plate for 30 minutes to minimize bacterial transfer from lawn.…”

Section: Gut Colonization Assaymentioning

confidence: 99%

Kombucha Tea-associated Microbes Remodel Host Metabolic Pathways to Suppress Lipid Accumulation

DuMez-Kornegay,

Morris,

Dowen

2023

Preprint

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“…elegans is a well-developed model for studying hostpathogen interactions and addressing several biological issues, such as aging, diabetes, wound mechanism, longevity, multigenerational adaptation, neurological disorders and DNA repairing mechanisms, owing to its small size, short life span, bacterivorus property, easy maintenance, transparency and complete genome sequencing. [14][15][16][17][18] Moreover, it has been established to apply the multi-OMICS approaches 19 for understanding complicated biological processes. The host-pathogen interaction between C. elegans and bacterial pathogens is based on the source of nutrient, aversive learning and host resistance, 20 which determine the longevity, immune response and other molecular regulations of the host.…”

Section: Introductionmentioning

confidence: 99%

Distinct global metabolomic profiles of the model organismCaenorhabditis elegansduring interactions withStaphylococcus aureusandSalmonella entericaSerovar Typhi

2023

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“…In laboratory experiments, it is sometimes desirable to pool multiple samples when the amount of biological material generated per sample is small (Kendziorski et al 2005;Mary-Huard et al 2007). For this reason, batch-based measurements are common in studies of hostmicrobe associations in small organisms such as the nematode Caenorhabditis elegans (Walker et al 2022). In these experiments, "batches" are created by collecting groups of individual hosts (usually 5-50 hosts per batch) into individual volumes for mechanical disruption and quantification, often by plating on solid agar to determine colony forming unit (CFU) counts.…”

Section: Introductionmentioning

confidence: 99%

“…2007). For this reason, batch-based measurements are common in studies of host-microbe associations in small organisms such as the nematode Caenorhabditis elegans (Walker et al . 2022).…”

Section: Introductionmentioning

confidence: 99%

Sample pooling inflates error rates in between-sample comparisons: an empirical investigation of the statistical properties of count-based data

Vega

Taylor

2022

Preprint

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Heterogeneity is ubiquitous across individuals in biological data, and sample batching, a form of biological averaging, inevitably loses information about this heterogeneity. The consequences for inference from biologically averaged data are frequently opaque. Here we investigate a case where biological averaging is common - count-based measurement of bacterial load in individual Caenorhabditis elegans - to empirically determine the consequences of batching. We find that both central measures and measures of variation on individual-based data contain biologically relevant information that is useful for distinguishing between groups, and that batch-based inference readily produces both false positive and false negative results in these comparisons. These results support the use of individual rather than batched samples when possible, illustrate the importance of understanding the distributions of individual-based data in experimental systems, and indicate the need to consider effect size when drawing conclusions from these data.

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Quantification of Bacterial Loads in Caenorhabditis elegans

Cited by 5 publications

References 7 publications

Kombucha Tea-associated Microbes Remodel Host Metabolic Pathways to Suppress Lipid Accumulation

Kombucha Tea-associated Microbes Remodel Host Metabolic Pathways to Suppress Lipid Accumulation

Distinct global metabolomic profiles of the model organismCaenorhabditis elegansduring interactions withStaphylococcus aureusandSalmonella entericaSerovar Typhi

Sample pooling inflates error rates in between-sample comparisons: an empirical investigation of the statistical properties of count-based data

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