Understanding When Poisson Log-Normal Models Outperform Penalized Poisson Regression for Microbiome Count Data

Multivariate count models are often justified by their ability to capture latent dependence, but researchers receive little guidance on when this added structure improves on simpler penalized marginal Poisson regression. We study this question using real microbiome data under a unified held-out evaluation framework. For count prediction, we compare PLN and GLMNet(Poisson) on 20 datasets spanning 32 to 18,270 samples and 24 to 257 taxa, using held-out Poisson deviance under leave-one-taxon-out prediction with 3-fold sample cross-validation rather than synthetic or in-sample criteria. For network inference, we compare PLNNetwork and GLMNet(Poisson) neighborhood selection on five publicly available datasets with experimentally validated microbial interaction truth. PLN outperforms GLMNet(Poisson) on most count-prediction datasets, with gains up to 38 percent. The primary predictor of the winner is the sample-to-taxon ratio, with mean absolute correlation as the strongest secondary signal and overdispersion as an additional predictor. PLNNetwork performs best on broad undirected interaction benchmarks, whereas GLMNet(Poisson) is better aligned with local or directional effects. Taken together, these results provide guidance for choosing between latent multivariate count models and penalized Poisson regression in biological count prediction and interaction recovery.

Fused Lasso Improves Accuracy of Co-occurrence Network Inference in Grouped Samples

Co-occurrence network inference algorithms have significantly advanced our understanding of microbiome communities. However, these algorithms typically analyze microbial associations within samples collected from a single environmental niche, often capturing only static snapshots rather than dynamic microbial processes. Previous studies have commonly grouped samples from different environmental niches together without fully considering how microbial communities adapt their associations when faced with varying ecological conditions. Our study addresses this limitation by explicitly investigating both spatial and temporal dynamics of microbial communities. We analyzed publicly available microbiome abundance data across multiple locations and time points, to evaluate algorithm performance in predicting microbial associations using our proposed Same-All Cross-validation (SAC) framework. SAC evaluates algorithms in two distinct scenarios: training and testing within the same environmental niche (Same), and training and testing on combined data from multiple environmental niches (All). To overcome the limitations of conventional algorithms, we propose fuser, an algorithm that, while not entirely new in machine learning, is novel for microbiome community network inference. It retains subsample-specific signals while simultaneously sharing relevant information across environments during training. Unlike standard approaches that infer a single generalized network from combined data, fuser generates distinct, environment-specific predictive networks. Our results demonstrate that fuser achieves comparable predictive performance to existing algorithms such as glmnet when evaluated within homogeneous environments (Same), and notably reduces test error compared to baseline algorithms in cross-environment (All) scenarios.

Cross-Validation for Training and Testing Co-occurrence Network Inference Algorithms

Microorganisms are found in almost every environment, including the soil, water, air, and inside other organisms, like animals and plants. While some microorganisms cause diseases, most of them help in biological processes such as decomposition, fermentation and nutrient cycling. A lot of research has gone into studying microbial communities in various environments and how their interactions and relationships can provide insights into various diseases. Co-occurrence network inference algorithms help us understand the complex associations of micro-organisms, especially bacteria. Existing network inference algorithms employ techniques such as correlation, regularized linear regression, and conditional dependence, which have different hyper-parameters that determine the sparsity of the network. Previous methods for evaluating the quality of the inferred network include using external data, and network consistency across sub-samples, both which have several drawbacks that limit their applicability in real microbiome composition data sets. We propose a novel cross-validation method to evaluate co-occurrence network inference algorithms, and new methods for applying existing algorithms to predict on test data. Our empirical study shows that the proposed method is useful for hyper-parameter selection (training) and comparing the quality of the inferred networks between different algorithms (testing).