From 5e581831c5049afe3d4bd78c0942eb39c98794cd Mon Sep 17 00:00:00 2001 From: Naomi Wilson Date: Thu, 18 Jul 2024 14:17:50 -0500 Subject: [PATCH 1/2] Update README to switch resistant/susceptible --- README.Rmd | 22 +- README.md | 593 ++++++++++--------- tools/readme/weiss_healthy6-1.png | Bin 11860 -> 64096 bytes tools/readme/weiss_healthy6_prediction-1.png | Bin 13034 -> 73733 bytes tools/readme/weiss_vancomycin_index-1.png | Bin 6655 -> 23604 bytes 5 files changed, 308 insertions(+), 307 deletions(-) diff --git a/README.Rmd b/README.Rmd index 318f5c2..df66980 100644 --- a/README.Rmd +++ b/README.Rmd @@ -90,13 +90,13 @@ result the lineage. Our goal will be to use the lineage information to predict which bacteria in each sample would be susceptible or resistant to various antibiotics. For a given antibiotic, we construct an *antibiotic-specific* Mircrobiome Response -Index by taking the log-ratio of the abundance for resistant organisms over -that of susceptible organisms. +Index by taking the log-ratio of the abundance for susceptible organisms over +that of resistant organisms. If the bacterial community is dominated by susceptible organisms, the index -will be negative. Conversely, the index is positive if susceptible organisms +will be positive Conversely, the index is negative if susceptible organisms constitute a minority. If an antibiotic has the predicted effect on a -bacterial community, the index should increase after the antibiotic is +bacterial community, the index should decrease after the antibiotic is introduced. Let's compute the vancomycin response index for each sample in the study. @@ -113,8 +113,8 @@ weiss2021_vanc %>% labs(x = "Study window", y = "Vancomycin-specific index") ``` -For healthy children, the medain value of the index is about 0.2, whereas it is -roughly 0.85 across the samples from children with sepsis. Let's look further +For healthy children, the medain value of the index is about -0.2, whereas it is +roughly -0.85 across the samples from children with sepsis. Let's look further into how the index was calculated, and check out which bacteria were labeled as susceptible or resistant to vancomycin. @@ -215,19 +215,19 @@ healthy6_data %>% Most taxa in the sample are annotated as susceptible to vancomycin, including the most abundant taxon, *Ruminococcaceae*. One taxon, RF39, is not annotated. Only a few taxa are annotated as resistant to vancomycin, thus it's not -surprising that the vancomycin index for the sample is negative. +surprising that the vancomycin index for the sample is posgative. ```{r} healthy6_data %>% summarise(vanc = mirix_vancomycin(proportion, lineage)) ``` How would we expect the proportions to change if the index increased to a -positive value, say 0.5? We can use `predict_abundance()` to run the +negative value, say -0.5? We can use `predict_abundance()` to run the calculation. ```{r weiss_healthy6_prediction} healthy6_data %>% - mutate(predicted = predict_abundance(0.5, proportion, susceptibility)) %>% + mutate(predicted = predict_abundance(-0.5, proportion, susceptibility)) %>% rename(observed = proportion) %>% pivot_longer( c(observed, predicted), names_to = "method", values_to = "abundance") %>% @@ -244,11 +244,11 @@ For susceptible taxa, the abundances have decreased. For the taxon that's not annotated, RF39 (near the middle), the abundance has not changed at all. To finish, let's re-calculate the vancomycin index for our predicted abundances, -so we can verify that it has the expected value of 0.5. +so we can verify that it has the expected value of -0.5. ```{r} healthy6_data %>% - mutate(predicted = predict_abundance(0.5, proportion, susceptibility)) %>% + mutate(predicted = predict_abundance(-0.5, proportion, susceptibility)) %>% summarise(vanc = mirix_vancomycin(predicted, lineage)) ``` diff --git a/README.md b/README.md index 0c62245..815896e 100644 --- a/README.md +++ b/README.md @@ -1,296 +1,297 @@ - - - -# mirix - - - -[![R-CMD-check](https://github.com/PennChopMicrobiomeProgram/mirix/actions/workflows/R-CMD-check.yaml/badge.svg)](https://github.com/PennChopMicrobiomeProgram/mirix/actions/workflows/R-CMD-check.yaml) -[![codecov](https://codecov.io/gh/PennChopMicrobiomeProgram/mirix/graph/badge.svg?token=7DBBaNoGDc)](https://codecov.io/gh/PennChopMicrobiomeProgram/mirix) - - -The goal of mirix is to calculate the Microbiome Response Index (MiRIx) -for a given bacterial community. Often this will be used to predict the -community’s susceptibility to an antibiotic. - -## Installation - -You can install the development version of mirix with `devtools`: - -``` r -#install.packages("devtools") -devtools::install_github("PennChopMicrobiomeProgram/mirix") -``` - -## Calculating antibiotic index values - -Here, we’ll load the `mirix` package, demonstrate some basic tasks, and -give some pointers for its use. Although we could accomplish all our -tasks in base R, the `mirix` package works very nicely with functions -from the tidyverse. We’ll import those functions now. - -``` r -library(tidyverse) -``` - -Next, we’ll load the `mirix` library. - -``` r -library(mirix) -``` - -This package comes with a built-in data set, `weiss2021_data`, from a -study of the gut microbiota in children with sepsis (PMID 33786436). The -children in this study were exposed to a range of antibiotics, and fecal -samples were collected across a series of time windows after diagnosis -(`study_window` A-E). Bacterial communities in the fecal samples were -profiled by sequencing the 16S rRNA gene, which serves as a fingerprint -to identify bacteria in each sample. The study included samples from a -set of healthy children in a similar age group for comparison. Here is a -summary of the number of samples in each time window: - -``` r -weiss2021_data %>% - distinct(sample_id, study_group, study_window) %>% - count(study_group, study_window) %>% - knitr::kable() -``` - -| study_group | study_window | n | -|:------------|:-------------|----:| -| Healthy | NA | 44 | -| Sepsis | A | 17 | -| Sepsis | B | 9 | -| Sepsis | C | 7 | -| Sepsis | D | 2 | -| Sepsis | E | 8 | - -for each sample collected, the data frame contains the relative -abundance of all bacteria with a proportion of more than 0.001. Here is -one of the sepsis samples, from subject 19019 in time window A. In this -sample, the sequencing results indicated the following types of -bacteria: - -``` r -weiss2021_data %>% - filter(sample_id %in% "Sepsis.19019.A") %>% - select(lineage, proportion) %>% - knitr::kable() -``` - -| lineage | proportion | -|:---------------------------------------------------------------------------------------------------------------------------------|-----------:| -| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Lactobacillales; f\_\_Enterococcaceae; g\_\_Enterococcus | 0.5491550 | -| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Lactobacillales; f\_\_Lactobacillaceae | 0.3418542 | -| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Pseudomonadales; f\_\_Pseudomonadaceae; g\_\_Pseudomonas | 0.0825587 | -| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Enterobacteriales; f\_\_Enterobacteriaceae | 0.0094255 | -| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Xanthomonadales; f\_\_Xanthomonadaceae; g\_\_Stenotrophomonas | 0.0069635 | -| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Bacillales; f\_\_Staphylococcaceae; g\_\_Staphylococcus | 0.0051231 | -| k\_\_Bacteria; p\_\_Actinobacteria; c\_\_Actinobacteria; o\_\_Actinomycetales; f\_\_Propionibacteriaceae | 0.0027708 | - -For each type of bacteria, the taxonomic assignment includes all the -taxon names from the kingdom on down to the most specific taxon that -could be determined from the sequence, in most cases the genus. -Following the [NCBI taxonomy -browser](https://www.ncbi.nlm.nih.gov/taxonomy), we call this result the -lineage. - -Our goal will be to use the lineage information to predict which -bacteria in each sample would be susceptible or resistant to various -antibiotics. For a given antibiotic, we construct an -*antibiotic-specific* Mircrobiome Response Index by taking the log-ratio -of the abundance for resistant organisms over that of susceptible -organisms. - -If the bacterial community is dominated by susceptible organisms, the -index will be negative. Conversely, the index is positive if susceptible -organisms constitute a minority. If an antibiotic has the predicted -effect on a bacterial community, the index should increase after the -antibiotic is introduced. - -Let’s compute the vancomycin response index for each sample in the -study. - -``` r -weiss2021_vanc <- weiss2021_data %>% - group_by(sample_id, study_group, study_window) %>% - summarise(vanc = mirix_vancomycin(proportion, lineage), .groups = "drop") - -weiss2021_vanc %>% - ggplot(aes(x=study_window, y = vanc)) + - geom_boxplot() + - facet_grid(~ study_group, scales = "free_x", space = "free_x") + - labs(x = "Study window", y = "Vancomycin-specific index") -``` - -![](tools/readme/weiss_vancomycin_index-1.png) - -For healthy children, the medain value of the index is about 0.2, -whereas it is roughly 0.85 across the samples from children with sepsis. -Let’s look further into how the index was calculated, and check out -which bacteria were labeled as susceptible or resistant to vancomycin. - -## Susceptible and resistant bacteria - -The `mirix` library offers some lower-level functions that show more -details about how the index was calculated. For each antibiotic with an -index function, there is an accompanying function to determine the -susceptibility for each lineage. Let’s list out the susceptibility for -the lineages in the sample from subject 19019: - -``` r -weiss2021_data %>% - filter(sample_id %in% "Sepsis.19019.A") %>% - mutate(susceptibility = antibiotic_susceptibility_vancomycin(lineage)) %>% - select(lineage, susceptibility) %>% - knitr::kable() -``` - -| lineage | susceptibility | -|:---------------------------------------------------------------------------------------------------------------------------------|:---------------| -| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Lactobacillales; f\_\_Enterococcaceae; g\_\_Enterococcus | susceptible | -| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Lactobacillales; f\_\_Lactobacillaceae | susceptible | -| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Pseudomonadales; f\_\_Pseudomonadaceae; g\_\_Pseudomonas | resistant | -| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Enterobacteriales; f\_\_Enterobacteriaceae | resistant | -| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Xanthomonadales; f\_\_Xanthomonadaceae; g\_\_Stenotrophomonas | resistant | -| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Bacillales; f\_\_Staphylococcaceae; g\_\_Staphylococcus | susceptible | -| k\_\_Bacteria; p\_\_Actinobacteria; c\_\_Actinobacteria; o\_\_Actinomycetales; f\_\_Propionibacteriaceae | susceptible | - -Looking down the list of taxa, we can see that three of the seven are -from the *Firmicutes* phylum, in which the organisms are generally -characterized by a Gram-positive cell wall structure. Vancomycin’s -mechanism of action is to disrupt construction of the cell wall in -Gram-positive organisms, so we would generally predict that organisms in -this phylum would be susceptible to the antibiotic. Likewise, the -*Actinobacteria* are Gram-positive, so the final lineage is predicted to -be susceptible. The three remaining lineages are Gram-negative organisms -from the *Proteobacteria*, and are predicted to be resistant. - -Now, if you’ve read the Wikipedia article on vancomycin, you might know -that most species of *Lactobacillus* are naturally resistant to the -drug, although they are Gram-positive bacteria in the *Firmicutes* -phylum. Here, we don’t have an assignment to the *Lactobacillus* genus, -but instead to the *Lactobacillaceae* family, so the prediction stands -as susceptible. - -## Databases for bacterial phenotypes and antibiotic susceptibility - -The `mirix` package comes with two built-in databases. The -`taxon_phenotypes` database contains information on Gram-stain and -aerobic status for over 900 bacterial taxa. The taxa were selected to -cover many taxa encountered in the human microbiome. It is here where we -note that the *Firmicutes* are generally Gram-positive. - -``` r -whatbacteria::taxon_phenotypes %>% - filter(taxon %in% "Firmicutes") -#> taxon rank aerobic_status gram_stain doi -#> 1 Firmicutes Phylum Gram-positive 10.1099/00207713-28-1-1 -``` - -The second database `taxon_susceptibility`, contains information about -bacterial susceptibility to specific antibiotics. It is here, for -example, where we note that *Lactobacillus* species are typically -resistant to vancomycin. - -``` r -whatbacteria::taxon_susceptibility %>% - filter(taxon %in% "Lactobacillus") -#> taxon rank antibiotic value doi -#> 1 Lactobacillus Genus vancomycin resistant 10.1128/AEM.01738-18 -``` - -If you need to add or modify information in these databases, you can -copy the data frames to new variables, make the changes you’d like, and -pass the new databases directly to the `mirix_vancomycin()` or -`antibiotic_susceptibility_vancomycin()` functions. - -## Predicting taxon abundances for a given value of the index - -In addition to calculating the antibiotics index for a given sample, we -can predict what the abundances in a sample might look like at a given -value of the index. Our approach is to re-balance the total abundances -of resistant and susceptible bacteria in the sample, while preserving -the relative abundances within the resistant taxa and within the -susceptible taxa. Our method will also preserve the total abundance of -each sample, in case the total abundance is not equal to 1. For taxa -that are not annotated as either resistant or susceptible, we don’t -change the abundance at all. - -We’ll use a healthy control sample, `Healthy.6`, to demonstrate. First, -we’ll calculate the susceptibility to vancomycin for each lineage. - -``` r -healthy6_data <- weiss2021_data %>% - filter(sample_id %in% "Healthy.6") %>% - mutate(taxon = word(lineage, -1)) %>% - mutate(taxon = fct_reorder(taxon, proportion)) %>% - mutate(susceptibility = antibiotic_susceptibility_vancomycin(lineage)) -``` - -Above, we also made a shorter label for the taxa and sorted the values -based on proportion in the sample, to aid in plotting. Here is a chart -of the taxon abundances and their susceptibility. - -``` r -healthy6_data %>% - ggplot(aes(x = proportion, y = taxon, shape = susceptibility)) + - geom_point() + - scale_x_log10() + - scale_shape_manual(values = c(1, 19), na.value = 3) -``` - -![](tools/readme/weiss_healthy6-1.png) - -Most taxa in the sample are annotated as susceptible to vancomycin, -including the most abundant taxon, *Ruminococcaceae*. One taxon, RF39, -is not annotated. Only a few taxa are annotated as resistant to -vancomycin, thus it’s not surprising that the vancomycin index for the -sample is negative. - -``` r -healthy6_data %>% - summarise(vanc = mirix_vancomycin(proportion, lineage)) -#> # A tibble: 1 × 1 -#> vanc -#> -#> 1 0.207 -``` - -How would we expect the proportions to change if the index increased to -a positive value, say 0.5? We can use `predict_abundance()` to run the -calculation. - -``` r -healthy6_data %>% - mutate(predicted = predict_abundance(0.5, proportion, susceptibility)) %>% - rename(observed = proportion) %>% - pivot_longer( - c(observed, predicted), names_to = "method", values_to = "abundance") %>% - ggplot(aes(x = abundance, y = taxon, color = method, shape = susceptibility)) + - geom_point() + - scale_x_log10() + - scale_shape_manual(values = c(1, 19), na.value = 3) + - scale_color_brewer(palette = "Paired") -``` - -![](tools/readme/weiss_healthy6_prediction-1.png) - -Here, we can see that the abundances have increased for the resistant -taxa, such as *Bacteroides* (near the top) and *Enterobacteriaceae* (at -the bottom). For susceptible taxa, the abundances have decreased. For -the taxon that’s not annotated, RF39 (near the middle), the abundance -has not changed at all. - -To finish, let’s re-calculate the vancomycin index for our predicted -abundances, so we can verify that it has the expected value of 0.5. - -``` r -healthy6_data %>% - mutate(predicted = predict_abundance(0.5, proportion, susceptibility)) %>% - summarise(vanc = mirix_vancomycin(predicted, lineage)) -#> # A tibble: 1 × 1 -#> vanc -#> -#> 1 0.50 -``` + + + +# mirix + + + +[![R-CMD-check](https://github.com/PennChopMicrobiomeProgram/mirix/actions/workflows/R-CMD-check.yaml/badge.svg)](https://github.com/PennChopMicrobiomeProgram/mirix/actions/workflows/R-CMD-check.yaml) +[![codecov](https://codecov.io/gh/PennChopMicrobiomeProgram/mirix/graph/badge.svg?token=7DBBaNoGDc)](https://codecov.io/gh/PennChopMicrobiomeProgram/mirix) + + +The goal of mirix is to calculate the Microbiome Response Index (MiRIx) +for a given bacterial community. Often this will be used to predict the +community’s susceptibility to an antibiotic. + +## Installation + +You can install the development version of mirix with `devtools`: + +``` r +#install.packages("devtools") +devtools::install_github("PennChopMicrobiomeProgram/mirix") +``` + +## Calculating antibiotic index values + +Here, we’ll load the `mirix` package, demonstrate some basic tasks, and +give some pointers for its use. Although we could accomplish all our +tasks in base R, the `mirix` package works very nicely with functions +from the tidyverse. We’ll import those functions now. + +``` r +library(tidyverse) +``` + +Next, we’ll load the `mirix` library. + +``` r +library(mirix) +``` + +This package comes with a built-in data set, `weiss2021_data`, from a +study of the gut microbiota in children with sepsis (PMID 33786436). The +children in this study were exposed to a range of antibiotics, and fecal +samples were collected across a series of time windows after diagnosis +(`study_window` A-E). Bacterial communities in the fecal samples were +profiled by sequencing the 16S rRNA gene, which serves as a fingerprint +to identify bacteria in each sample. The study included samples from a +set of healthy children in a similar age group for comparison. Here is a +summary of the number of samples in each time window: + +``` r +weiss2021_data %>% + distinct(sample_id, study_group, study_window) %>% + count(study_group, study_window) %>% + knitr::kable() +``` + +| study_group | study_window | n | +|:------------|:-------------|----:| +| Healthy | NA | 44 | +| Sepsis | A | 17 | +| Sepsis | B | 9 | +| Sepsis | C | 7 | +| Sepsis | D | 2 | +| Sepsis | E | 8 | + +for each sample collected, the data frame contains the relative +abundance of all bacteria with a proportion of more than 0.001. Here is +one of the sepsis samples, from subject 19019 in time window A. In this +sample, the sequencing results indicated the following types of +bacteria: + +``` r +weiss2021_data %>% + filter(sample_id %in% "Sepsis.19019.A") %>% + select(lineage, proportion) %>% + knitr::kable() +``` + +| lineage | proportion | +|:---------------------------------------------------------------------------------------------------------------------------------|-----------:| +| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Lactobacillales; f\_\_Enterococcaceae; g\_\_Enterococcus | 0.5491550 | +| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Lactobacillales; f\_\_Lactobacillaceae | 0.3418542 | +| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Pseudomonadales; f\_\_Pseudomonadaceae; g\_\_Pseudomonas | 0.0825587 | +| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Enterobacteriales; f\_\_Enterobacteriaceae | 0.0094255 | +| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Xanthomonadales; f\_\_Xanthomonadaceae; g\_\_Stenotrophomonas | 0.0069635 | +| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Bacillales; f\_\_Staphylococcaceae; g\_\_Staphylococcus | 0.0051231 | +| k\_\_Bacteria; p\_\_Actinobacteria; c\_\_Actinobacteria; o\_\_Actinomycetales; f\_\_Propionibacteriaceae | 0.0027708 | + +For each type of bacteria, the taxonomic assignment includes all the +taxon names from the kingdom on down to the most specific taxon that +could be determined from the sequence, in most cases the genus. +Following the [NCBI taxonomy +browser](https://www.ncbi.nlm.nih.gov/taxonomy), we call this result the +lineage. + +Our goal will be to use the lineage information to predict which +bacteria in each sample would be susceptible or resistant to various +antibiotics. For a given antibiotic, we construct an +*antibiotic-specific* Mircrobiome Response Index by taking the log-ratio +of the abundance for susceptible organisms over that of resistant +organisms. + +If the bacterial community is dominated by susceptible organisms, the +index will be positive Conversely, the index is negative if susceptible +organisms constitute a minority. If an antibiotic has the predicted +effect on a bacterial community, the index should decrease after the +antibiotic is introduced. + +Let’s compute the vancomycin response index for each sample in the +study. + +``` r +weiss2021_vanc <- weiss2021_data %>% + group_by(sample_id, study_group, study_window) %>% + summarise(vanc = mirix_vancomycin(proportion, lineage), .groups = "drop") + +weiss2021_vanc %>% + ggplot(aes(x=study_window, y = vanc)) + + geom_boxplot() + + facet_grid(~ study_group, scales = "free_x", space = "free_x") + + labs(x = "Study window", y = "Vancomycin-specific index") +``` + +![](tools/readme/weiss_vancomycin_index-1.png) + +For healthy children, the medain value of the index is about -0.2, +whereas it is roughly -0.85 across the samples from children with +sepsis. Let’s look further into how the index was calculated, and check +out which bacteria were labeled as susceptible or resistant to +vancomycin. + +## Susceptible and resistant bacteria + +The `mirix` library offers some lower-level functions that show more +details about how the index was calculated. For each antibiotic with an +index function, there is an accompanying function to determine the +susceptibility for each lineage. Let’s list out the susceptibility for +the lineages in the sample from subject 19019: + +``` r +weiss2021_data %>% + filter(sample_id %in% "Sepsis.19019.A") %>% + mutate(susceptibility = antibiotic_susceptibility_vancomycin(lineage)) %>% + select(lineage, susceptibility) %>% + knitr::kable() +``` + +| lineage | susceptibility | +|:---------------------------------------------------------------------------------------------------------------------------------|:---------------| +| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Lactobacillales; f\_\_Enterococcaceae; g\_\_Enterococcus | susceptible | +| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Lactobacillales; f\_\_Lactobacillaceae | susceptible | +| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Pseudomonadales; f\_\_Pseudomonadaceae; g\_\_Pseudomonas | resistant | +| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Enterobacteriales; f\_\_Enterobacteriaceae | resistant | +| k\_\_Bacteria; p\_\_Proteobacteria; c\_\_Gammaproteobacteria; o\_\_Xanthomonadales; f\_\_Xanthomonadaceae; g\_\_Stenotrophomonas | resistant | +| k\_\_Bacteria; p\_\_Firmicutes; c\_\_Bacilli; o\_\_Bacillales; f\_\_Staphylococcaceae; g\_\_Staphylococcus | susceptible | +| k\_\_Bacteria; p\_\_Actinobacteria; c\_\_Actinobacteria; o\_\_Actinomycetales; f\_\_Propionibacteriaceae | susceptible | + +Looking down the list of taxa, we can see that three of the seven are +from the *Firmicutes* phylum, in which the organisms are generally +characterized by a Gram-positive cell wall structure. Vancomycin’s +mechanism of action is to disrupt construction of the cell wall in +Gram-positive organisms, so we would generally predict that organisms in +this phylum would be susceptible to the antibiotic. Likewise, the +*Actinobacteria* are Gram-positive, so the final lineage is predicted to +be susceptible. The three remaining lineages are Gram-negative organisms +from the *Proteobacteria*, and are predicted to be resistant. + +Now, if you’ve read the Wikipedia article on vancomycin, you might know +that most species of *Lactobacillus* are naturally resistant to the +drug, although they are Gram-positive bacteria in the *Firmicutes* +phylum. Here, we don’t have an assignment to the *Lactobacillus* genus, +but instead to the *Lactobacillaceae* family, so the prediction stands +as susceptible. + +## Databases for bacterial phenotypes and antibiotic susceptibility + +The `mirix` package comes with two built-in databases. The +`taxon_phenotypes` database contains information on Gram-stain and +aerobic status for over 900 bacterial taxa. The taxa were selected to +cover many taxa encountered in the human microbiome. It is here where we +note that the *Firmicutes* are generally Gram-positive. + +``` r +whatbacteria::taxon_phenotypes %>% + filter(taxon %in% "Firmicutes") +#> taxon rank aerobic_status gram_stain doi +#> 1 Firmicutes Phylum Gram-positive 10.1099/00207713-28-1-1 +``` + +The second database `taxon_susceptibility`, contains information about +bacterial susceptibility to specific antibiotics. It is here, for +example, where we note that *Lactobacillus* species are typically +resistant to vancomycin. + +``` r +whatbacteria::taxon_susceptibility %>% + filter(taxon %in% "Lactobacillus") +#> taxon rank antibiotic value doi +#> 1 Lactobacillus Genus vancomycin resistant 10.1128/AEM.01738-18 +``` + +If you need to add or modify information in these databases, you can +copy the data frames to new variables, make the changes you’d like, and +pass the new databases directly to the `mirix_vancomycin()` or +`antibiotic_susceptibility_vancomycin()` functions. + +## Predicting taxon abundances for a given value of the index + +In addition to calculating the antibiotics index for a given sample, we +can predict what the abundances in a sample might look like at a given +value of the index. Our approach is to re-balance the total abundances +of resistant and susceptible bacteria in the sample, while preserving +the relative abundances within the resistant taxa and within the +susceptible taxa. Our method will also preserve the total abundance of +each sample, in case the total abundance is not equal to 1. For taxa +that are not annotated as either resistant or susceptible, we don’t +change the abundance at all. + +We’ll use a healthy control sample, `Healthy.6`, to demonstrate. First, +we’ll calculate the susceptibility to vancomycin for each lineage. + +``` r +healthy6_data <- weiss2021_data %>% + filter(sample_id %in% "Healthy.6") %>% + mutate(taxon = word(lineage, -1)) %>% + mutate(taxon = fct_reorder(taxon, proportion)) %>% + mutate(susceptibility = antibiotic_susceptibility_vancomycin(lineage)) +``` + +Above, we also made a shorter label for the taxa and sorted the values +based on proportion in the sample, to aid in plotting. Here is a chart +of the taxon abundances and their susceptibility. + +``` r +healthy6_data %>% + ggplot(aes(x = proportion, y = taxon, shape = susceptibility)) + + geom_point() + + scale_x_log10() + + scale_shape_manual(values = c(1, 19), na.value = 3) +``` + +![](tools/readme/weiss_healthy6-1.png) + +Most taxa in the sample are annotated as susceptible to vancomycin, +including the most abundant taxon, *Ruminococcaceae*. One taxon, RF39, +is not annotated. Only a few taxa are annotated as resistant to +vancomycin, thus it’s not surprising that the vancomycin index for the +sample is posgative. + +``` r +healthy6_data %>% + summarise(vanc = mirix_vancomycin(proportion, lineage)) +#> # A tibble: 1 × 1 +#> vanc +#> +#> 1 0.207 +``` + +How would we expect the proportions to change if the index increased to +a negative value, say -0.5? We can use `predict_abundance()` to run the +calculation. + +``` r +healthy6_data %>% + mutate(predicted = predict_abundance(-0.5, proportion, susceptibility)) %>% + rename(observed = proportion) %>% + pivot_longer( + c(observed, predicted), names_to = "method", values_to = "abundance") %>% + ggplot(aes(x = abundance, y = taxon, color = method, shape = susceptibility)) + + geom_point() + + scale_x_log10() + + scale_shape_manual(values = c(1, 19), na.value = 3) + + scale_color_brewer(palette = "Paired") +``` + +![](tools/readme/weiss_healthy6_prediction-1.png) + +Here, we can see that the abundances have increased for the resistant +taxa, such as *Bacteroides* (near the top) and *Enterobacteriaceae* (at +the bottom). For susceptible taxa, the abundances have decreased. For +the taxon that’s not annotated, RF39 (near the middle), the abundance +has not changed at all. + +To finish, let’s re-calculate the vancomycin index for our predicted +abundances, so we can verify that it has the expected value of -0.5. + +``` r +healthy6_data %>% + mutate(predicted = predict_abundance(-0.5, proportion, susceptibility)) %>% + summarise(vanc = mirix_vancomycin(predicted, lineage)) +#> # A tibble: 1 × 1 +#> vanc +#> +#> 1 -0.5 +``` diff --git a/tools/readme/weiss_healthy6-1.png b/tools/readme/weiss_healthy6-1.png index aa67b006c1d2bfc3a102655c290a27cc4deb8cab..33c86e9e947c2032aeaea9dad268063cbe395f98 100644 GIT binary patch literal 64096 zcmY&gbyO7H-zTL+Qcw_f=`QK+?h+-HkP?uR?nWAzytA=WfB(kFMB2pK#LnDEO~ zY9szLzwb;ct>zeG-&Ki5_3q54nNzc`+~k_m3E_)kw-qkNF5c_3(U4h`?0#g*u2RWH z@}s>NzLL%wB}ChC;Cs+@WdYS4pxG#aOf7wO5Vgyk*C%pK@d+s|Wr9V%=E+yYPJx zIkS0XC-%Kby*b(;1)N@EQ-B@%p-_jBrd!jq-ws8^+#}x@3dWO6u^0TD9oq!@y7nY) zs@V)u(H%Y2QXj5kk-{x5(=&Z>J*Oc3qG(PZ+#jxG#?^fkd{MVf@D{;ikIB@WZ)%t~ z?`^0k+#a6rpd)NsB!R$T%PMuoQ>^+&uGmcT2qOI#O~?}bv#ZC?-gnDciDN&`WjCn z3W{a!uJbyl0G;T!S=R47v#g7W#hr!kbeDH?-S1vK(j(O|QCfnDGdo+#}BJcyfE=>Bp!ZDV;=P?j!ua#e`LQ8}88Fm@A3*J3?i_CTgb_TW( z`wVG|ahH7#%f0fPujuRM3954$V%k5M1Za%|1i!a`Ho+-;K{k@*hnThz% zYxqUjo45wEb8v7XaIzAj8qV-LX()A%C+hDKdVky#85Ii_KFKqq2|`0TIuU3yp`k>oI7m1FPgISdJ-ropSo+Syot*3gN zO&9FmdJfqSo^V?JdgZpBR=#&!j$n&R21WAs#>Pev6G26VCsR>EbuUjH%ozi(|NDlx z2$BY(DC{@>Ihv`9gALE5Rp6eyB}2negF6U-Tu9r zQ_l(~5sQoaai#O-^EOi)1x_lY|6I9>00Mbtr%|rpe~v+sDuTUH%?R2w)c?Axf8VoC ziYRrVOr#t}otlB$DX-gv#?=`4(AKMAQhujB2d}HK3|z}YHi4Zl?Ye1vj&oi&(jPKi zw@Ygo-{-&mksNYtCiR~OaOp3iWG(`6UWBhT>_ks6mzcLKZ(b*L{Y_SDIz)Qy#~4Av zKdI+5@A&Ftg(tXS^EFnpN9JLh+5vx_Hnbc2WxZV<|K)1zJX`m(m1IuKx1kx&8YT3U zo>D}SRO>%=dY-o4dUs_R^x!y*eeg@GLqy0-f$Qm3nax5=h@yl@jpfu}^Z75gN2O~E zcW2z^E0Fu2(bEt1Twp@f)9cBR+aBp+RmnQ)PR- zQyF=kaeoPsHa^Glx)48K3W7BFJ>u)R{VgqA;c+88L;Y!xEa=&?)^uF({(Pg;EN{#G z?LJM7t@U7ceLK-?>&+U=W5{&1Wdi?h)!5ujwdH)v^`6e= zRmmewB_z8*Zy|845fEfyB$@fkjf{-P9b_t~l(?HS)g>uxMu9j?sB50#(i}!*tw(cZ<9{KFHF{is z;C4MRo5~bLJl`zHNM3(jfsrJ5^*d+2wD#8)G^Gx8?tMF^R3OV9+B-3X8PprfYJ`Oq+kI>|n})M_L< zfLOr!i{5pW`Pf_xmHf`fgmLLu8fC5@2brRNVJejo=XTJC)9ILJsQ-;MPNf+eFz_bYA9y&^GlZHe1qe_8bRw-n$s^^ z@t{Z5B$uM1v@DG~6;%UQW+(;98P>VccOr9%gDKoYg6DJgQX+nW-|)TeZ_gVIZnw1d z+E5@I-ygsSD3E$yEJ>-1#65b(Gu(W&RZ^y!BV~PYu(a1jY{M}sYXI>$Q5}mB^W7Ls zvoy#!-WYPiAYgni9{BL<-808=RqLN$-bmrTLTL-PD6n1WvR>&5-y7uLcYv+0a2}_v z@!lTtRnjBbM)#$`B=+v+i{+1xd?Vf_TCR~Heb~{dwVwZ3!#XcPqF%1wj3l5-jK$8P zrRS*Gm%vF$csOIyTAn}1aWX1F8Hp60U@YJ(@1APWgoI;zhVz36+$-BRAx?TPo$roY zMtRUFa@KfY3$9D=wd@q0!+48_o4mD<N zxrVp{pAsL1JhSbZo5a-k1T1KK#S|{Zn**Ji zcbg54+iyE?$b%*cvPwhnX!tk^RCm7&olY6-r+eMGpo9cKkJlp2Ay%}jzl3#JTcJ@# ztMNM4U+Jt7OSH;AMD>C-(ht^1ps$yi%;m8!0dMNdG$xXtMlmVzB zlm}Q-&U76eL*`A+l;{PXO_}~Gu^f6W9a*qG8=vC7qN*^y-wHdwddgl3p*TRi7 z+mhQGue+aw!b&Z2&9KE^BUDU+Y73snK-ROPxjcD7ij`-?)+o<^m*8b3byn>okFm@y z<8A8r>WwqyK>`ku^y;Zss`ho5Mrjfpo!HMyF4;Q(Lc0#GwV*i4YyLDC95FAvtjnj8*#Cf z({Xj>q!0KpA!jE6g!-%q7<8R+m}KD=C}RSi1sCTW=A)EZ(E&H@G5oqRTdL{2>#16zTQ$K>V}yPf71(% zCkS;uvi>6Zs5|Pf2XD3VQ8u=1qnKlUp{NsmLMBM8KdgT!?Pbqg^)a#2Ppp;ILt0XJ zjDXr5g0>U+(E_EE;aZY+!T!;m83QfSyCowcq5Q&{S#t~p}oI-;+0XT z1;gGGuh04aLcEAIc~oDA-69_wC!PX6mDzrupMX@F&i4N>13L}GP)N42O>PR~|Imnf zY!!U>wFbym_`h5XieyI$k29Rw+SwrTAC9HLjAS>8Qe`Ka`pQN|ebwF?cUMs^*C|%ZTeBg#GLldJ;C9Ew! 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ztCTe<+QO}eyjOFNK0XGt2x3RAgKpiY3kLWDq92Z)IPv*MmZ2{BcBw+e&ayWbw`dxL vZi>Bf+>8`_yRth{^&f}#-(=SyeVXAoV2X Date: Thu, 18 Jul 2024 14:22:59 -0500 Subject: [PATCH 2/2] typo --- README.Rmd | 2 +- README.md | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/README.Rmd b/README.Rmd index df66980..5122e16 100644 --- a/README.Rmd +++ b/README.Rmd @@ -113,7 +113,7 @@ weiss2021_vanc %>% labs(x = "Study window", y = "Vancomycin-specific index") ``` -For healthy children, the medain value of the index is about -0.2, whereas it is +For healthy children, the median value of the index is about -0.2, whereas it is roughly -0.85 across the samples from children with sepsis. Let's look further into how the index was calculated, and check out which bacteria were labeled as susceptible or resistant to vancomycin. diff --git a/README.md b/README.md index 815896e..60a414d 100644 --- a/README.md +++ b/README.md @@ -125,7 +125,7 @@ weiss2021_vanc %>% ![](tools/readme/weiss_vancomycin_index-1.png) -For healthy children, the medain value of the index is about -0.2, +For healthy children, the median value of the index is about -0.2, whereas it is roughly -0.85 across the samples from children with sepsis. Let’s look further into how the index was calculated, and check out which bacteria were labeled as susceptible or resistant to