25 Emerging infectious diseases are a growing threat to biodiversity worldwide. Outbreaks of the 26 infectious disease chytridiomycosis, caused by the fungal pathogen Batrachochytrium 27 dendrobatidis (Bd), have caused the decline and extinction of numerous amphibian species. In 28 Costa Rica, a major decline event occurred in 1987, more than two decades before this pathogen 29 was discovered. The loss of many species in Costa Rica is assumed to be due to Bd-epizootics, 30 but there are few studies that provide data from amphibians in the time leading up to the 31 proposed epizootics. In this study, we provide new data on Bd infection rates of amphibians 32 collected throughout Costa Rica, in the decades prior to the epizootics. We used a quantitative 33 PCR assay to test for Bd infection in 1016 specimens collected throughout Costa Rica. We found 34 Bd-infected hosts collected as early as 1964, and a infection prevalence average per decade of 35 just 4%. The infection prevalence remained relatively low and geographically constrained until 36 the 1980s when epizootics are hypothesized to have occurred. After that time, infection 37 prevalence increased three-fold and Bd-infected hosts we collected throughout the entire country.38 Our results, suggest that Bd may either have invaded Costa Rica earlier than previously known, 39 and spread more slowly than previously reported, or that an endemic lineage of the pathogen 40 may exists. To help visualize areas where future studies should take place, we provide a Bd 41 habitat suitability model trained with local data. Studies that provide information on genetic 42 lineages of Bd are needed to determine whether an endemic lineage of Bd or the Global 43 Panzootic Lineage (identified from mass die off sites globally) was present in Costa Rica and 44 responsible for the epizootics that caused amphibian communities to collapse. 45 46 Introduction 3 47 Amphibians are experiencing a global extinction event [1,2]. Though many factors contribute to 48 population declines, the emergence of the fungal pathogen Batrachochytrium dendrobatidis (Bd) 49 is one of the most important [3]. The disease chytridiomycosis, caused by the fungal pathogen 50 Batrachochytrium dendrobatidis (hereafter Bd), was first described in 1999 and has since been 51 found all over the world [3][4][5]. Interestingly, Bd is composed of many genetic lineages that vary 52 in virulence and affect host species differently. The panzootic disease is attributed to Bd-GPL, a 53 Global Panzootic Lineage of Bd associated with Bd epizootics and host population collapse [6].54 Other lineages of Bd have been shown to be less virulent and have been identified in areas 55 lacking epizootics [7]. Bd infects the skin of the amphibian and causes hyperkeratosis, the 56 thickening of skin which disrupts the amphibian's osmotic balance; leading to death by cardiac 57 arrest in highly infected individuals [8,9].
58The dynamics of Bd and its hosts, including pathogen invasion and the host-pathogen 59 interactions that follow, ar...