Commensal bacterial sharing does not predict host social associations in kangaroos.

Social network analysis has been postulated as a tool to study potential pathogen transmission in wildlife but is resource-intensive to quantify. Networks based on bacterial genotypes have been proposed as a cost-effective method for estimating social or transmission network based on the assumption that individuals in close contact will share commensal bacteria. However, the use of network analysis to study wild populations requires critical evaluation of the assumptions and parameters these models are founded on. We test (a) whether networks of commensal bacterial sharing are related to hosts' social associations and hence could act as a proxy for estimating transmission networks, (b) how the parameters chosen to define host associations and delineate bacterial genotypes impact inference and (c) whether these relationships change across time. We use stochastic simulations to evaluate how uncertainty in parameter choice affects network structure. We focused on a well-studied population of eastern grey kangaroos (Macropus giganteus), from Sundown National Park, Australia. Using natural markings, each individual was identified and its associations with other kangaroos recorded through direct field observations over 2 years to construct social networks. Faecal samples were collected, Escherichia coli was cultured and genotyped using BOX-PCR, and bacterial networks were constructed. Two individuals were connected in the bacterial network if they shared at least one E. coli genotype. We determined the capacity of bacterial networks to predict the observed social network structure in each year. We found little support for a relationship between social association and dyadic commensal bacterial similarity. Thresholds to determine host associations and similarity cut-off values used to define E. coli genotypes had important ramifications for inferring links between individuals. In fact, we found that inferences can show opposite patterns based on the chosen thresholds. Moreover, no similarity in overall bacterial network structure was detected between years. Although empirical disease transmission data are often unavailable in wildlife populations, both bacterial networks and social networks have limitations in representing the mode of transmission of a pathogen. Our results suggest that caution is needed when designing such studies and interpreting results.