Cryo-archiving of Batrachochytrium dendrobatidis and other chytridiomycetes

Batrachochytrium dendrobatidis is a major pathogen of frogs worldwide. It has been associated with catastrophic declines of frog populations including those in pristine habitats in Queensland, Australia. To facilitate genetic and disease studies of this fungus and related species, it is essential to have a reliable long-term storage method to maintain genetic integrity of isolates. We have adapted well-established techniques used for the long-term storage of tissue-culture cell lines to the preservation of B. dendrobatidis and other chytridiomycetes. This simple method has allowed us to recover these fungi from storage at -80 degrees C and in liquid nitrogen over an extended period. With this technique it is now possible to preserve saprobic and parasitic isolates from a variety of environmental and disease situations for comparative genetic and biological studies.     

Description of the oöcysts of theree new species ofEimeria (Apicomplexa: Eimeriidae) from geckoes (Auria: Gekkonidae)

Three new species ofEimeria are described from the faeces of geckoes. Oöcysts ofEimeria vittati n. sp. fromGekko vittatus are elongate-ellipsoid 34.3 × 16.9 (32.5–36.5 × 16.5–17.5) μm; shape index (mean length/mean width) 2.03. Its sporocysts are ovoid 11.0 × 6.5 (10–12.5 × 5–7.5) μ; shape index 1.7. Oöcysts ofE. simonkingi n. sp. were found inG. vitttus and two subspecies ofPhelsuma lineata, and are spherical to subspherical 20.5 × 19.4 (19.5–22 × 17.5–21) μm; shape index 1.06. Its sporocysts are ellipsoidal 9.9 × 5.9 (8–11.5 × 5.5–7) shape index 1.7. Oöcysts ofE. tokayae n. sp. fromG. gecko were spherical or subspherical, 18.3 × 18.2 (17–21 × 13–20.5) μm; shape index 1.01. Its sporocysts are ellipsoidal 9.2 × 5.9 (8–11.5 × 5.5–7) All of the new species ofEimeria from gekkonid lizards described since 1986 are listed in a reference table, and details of their oöcyst and sporocyst measurements and their morphology are given.     

Bridging Taxonomic and Disciplinary Divides in Infectious Disease

Pathogens traverse disciplinary and taxonomic boundaries, yet infectious disease research occurs in many separate disciplines including plant pathology, veterinary and human medicine, and ecological and evolutionary sciences. These disciplines have different traditions, goals, and terminology, creating gaps in communication. Bridging these disciplinary and taxonomic gaps promises novel insights and important synergistic advances in control of infectious disease. An approach integrated across the plant-animal divide would advance our understanding of disease by quantifying critical processes including transmission, community interactions, pathogen evolution, and complexity at multiple spatial and temporal scales. These advances require more substantial investment in basic disease research.     

Secretion of multilamellar whorls by Eimeria tenella zoites.

Multilamellar whorls were demonstrated by transmission electron microscopy to be associated with sporozoites and all generations of merozoites of Eimeria tenella, in chicken cecal tissue fixed without tannic acid or ruthenium red at room temperature. Whorls were found within the parasitophorous vacuoles of recently invaded cells at all stages of development, suggesting a role in the formation of the host parasite interface. Whorls were also associated with intraluminal third-generation merozoites prior to host cell invasion and appeared to be secreted directly through the pellicle. Membranous sheaths, shown by serial sectioning to be derived from intracellular whorl material, were observed enveloping some intraluminal merozoites. In many third-generation merozoites, whorl material was located within discrete novel organelles (here termed lamellosomes) located in the apical region. These densely staining spherical organelles were morphologically distinct from micronemes and rhoptries and were one-third the size of dense granules. These findings confirm that whorls are nonartifactual secretions whose lamellar organization is lost during normal fixation on ice without tannic acid. It is hypothesized that whorls secreted prior to invasion are involved in protection of the motile zoite, immune evasion, or some aspect of gliding motility.     

Identification of GBV-D,a Novel GB-like Flavivirus from Old World Frugivorous Bats (Pteropus giganteus) in Bangladesh

Bats are reservoirs for a wide range of zoonotic agents including lyssa-, henipah-, SARS-like corona-, Marburg-, Ebola-, and astroviruses. In an effort to survey for the presence of other infectious agents, known and unknown, we screened sera from 16 Pteropus giganteus bats from Faridpur, Bangladesh, using high-throughput pyrosequencing. Sequence analyses indicated the presence of a previously undescribed virus that has approximately 50% identity at the amino acid level to GB virus A and C (GBV-A and -C). Viral nucleic acid was present in 5 of 98 sera (5%) from a single colony of free-ranging bats. Infection was not associated with evidence of hepatitis or hepatic dysfunction. Phylogenetic analysis indicates that this first GBV-like flavivirus reported in bats constitutes a distinct species within the Flaviviridae family and is ancestral to the GBV-A and -C virus clades.