Background
Corynebacterium pseudotuberculosis, a Gram-positive, facultative intracellular pathogen, is the etiologic agent of the disease known as caseous lymphadenitis (CL). CL mainly affects small ruminants, such as goats and sheep; it also causes infections in humans, though rarely. This species is distributed worldwide, but it has the most serious economic impact in Oceania, Africa and South America. Although C. pseudotuberculosis causes major health and productivity problems for livestock, little is known about the molecular basis of its pathogenicity.Methodology and FindingsWe characterized two C. pseudotuberculosis genomes (Cp1002, isolated from goats; and CpC231, isolated from sheep). Analysis of the predicted genomes showed high similarity in genomic architecture, gene content and genetic order. When C. pseudotuberculosis was compared with other Corynebacterium species, it became evident that this pathogenic species has lost numerous genes, resulting in one of the smallest genomes in the genus. Other differences that could be part of the adaptation to pathogenicity include a lower GC content, of about 52%, and a reduced gene repertoire. The C. pseudotuberculosis genome also includes seven putative pathogenicity islands, which contain several classical virulence factors, including genes for fimbrial subunits, adhesion factors, iron uptake and secreted toxins. Additionally, all of the virulence factors in the islands have characteristics that indicate horizontal transfer.ConclusionsThese particular genome characteristics of C. pseudotuberculosis, as well as its acquired virulence factors in pathogenicity islands, provide evidence of its lifestyle and of the pathogenicity pathways used by this pathogen in the infection process. All genomes cited in this study are available in the NCBI Genbank database (http://www.ncbi.nlm.nih.gov/genbank/) under accession numbers CP001809 and CP001829.
Biomphalaria glabrata snails were experimentally infected with Angiostrongylus vasorum first-stage larvae and divided into four groups of 30 snails. To assess the shedding of third-stage larvae (L3), the snails were maintained under different stimuli: group 1 60 W light bulb for 24 h, group 2 37 degrees C water bath for 24 h, group 3 room temperature (23-25 degrees C) for 24 h, Group 4 room temperature (23-25 degrees C) for up to 15 days. After 24 h, a total of 512 A. vasorum L3, alive and active, were released by snails from group 1, while 2,446 L3 were released from group 2 and five L3 from group 3. After 15 days, snails from group 4 released a total of 44 L3. To evaluate the infectivity of A. vasorum L3, two mongrel dogs were successfully infected with L3 released by snails from groups 1 and 2, confirming that the infection of dogs with A. vasorum L3 was possible, independently of ingestion of the mollusk intermediate host. The results shown in these experiments suggest that angiostrongylosis could be directly transmitted to the definitive hosts, with implications for the parasite's life cycle.
An improved method to obtain a large number of axenic larvae of Angiostrongylus vasorum from fecal samples was developed in the present study. The procedure here in reported consisted of obtaining larvae using a modified Baermann technique, followed by an additional filtration step. This isolation technique recovered almost 90% of the living larvae in a clean preparation. Isolated larvae were submitted to decontamination treatments with either sodium hypochlorite or antibiotic cocktail solutions. The axenic status, as confirmed by oral inoculation of decontaminated larvae into germ-free mice, was only achieved using larvae treated with 0.5% sodium hypochlorite solution for 10 min. The isolation and decontamination treatment did not affect larval viability. Treated larvae remained viable and infective to the invertebrate host.
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