Interaction between Mycobacterium avium subsp. paratuberculosis and environmental protozoa

BackgroundInteractions between Mycobacterium avium subsp. paratuberculosis (Map) and free-living protozoa in water are likely to occur in nature. The potential impact of ingestion of Map by two naturally occurring Acanthamoeba spp. on this pathogen's survival and chlorine resistance was investigated.ResultsBetween 4.6 and 9.1% of spiked populations of three Map strains (NCTC 8578, B2 and ATCC 19698), which had been added at a multiplicity of infection of 10:1, were ingested by Acanthamoeba castellanii CCAP 1501/1B and A. polyphaga CCAP 1501/3B during co-culture for 3 h at 25°C. Map cells were observed to be present within the vacuoles of the amoebae by acid-fast staining. During extended co-culture of Map NCTC 8578 at 25°C for 24 d with both A. castellanii and A. polyphaga Map numbers did not change significantly during the first 7 days of incubation, however a 1–1.5 log10 increase in Map numbers was observed between days 7 and 24 within both Acanthamoeba spp. Ingested Map cells were shown to be more resistant to chlorine inactivation than free Map. Exposure to 2 μg/ml chlorine for 30 min resulted in a log10 reduction of 0.94 in ingested Map but a log10 reduction of 1.73 in free Map (p < 0.001).ConclusionThis study demonstrated that ingestion of Map by and survival and multiplication of Map within Acanthamoeba spp. is possible, and that Map cells ingested by amoebae are more resistant to inactivation by chlorine than free Map cells. These findings have implications with respect to the efficacy of chlorination applied to Map infected surface waters.

[1]  C. J. Clarke The pathology and pathogenesis of paratuberculosis in ruminants and other species. , 1997, Journal of comparative pathology.

[2]  A. E. Greenberg,et al.  Standard methods for the examination of water and wastewater. 14th edition. , 1976 .

[3]  Michael Wagner,et al.  Amoebae as Training Grounds for Intracellular Bacterial Pathogens , 2005, Applied and Environmental Microbiology.

[4]  R. Whittington,et al.  Use of Growth Indices from Radiometric Culture for Quantification of Sheep Strains of Mycobacterium avium subsp. paratuberculosis , 2003, Applied and Environmental Microbiology.

[5]  A Review of the Evidence for a Link between Exposure to Mycobacterium Paratuberculosis ( MAP ) and Crohn ’ s Disease ( CD ) in Humans , 2001 .

[6]  I. Grant,et al.  Bactericidal effect of chlorine on Mycobacterium paratuberculosis in drinking water , 2001, Letters in applied microbiology.

[7]  M. Rohde,et al.  Characterization of the intracellular survival of Mycobacterium avium ssp. paratuberculosis: phagosomal pH and fusogenicity in J774 macrophages compared with other mycobacteria , 2001, Cellular microbiology.

[8]  E. Shotts,et al.  Survival of coliforms and bacterial pathogens within protozoa during chlorination , 1988, Applied and environmental microbiology.

[9]  Richard J. Whittington,et al.  Survival and Dormancy of Mycobacterium aviumsubsp. paratuberculosis in theEnvironment , 2004, Applied and Environmental Microbiology.

[10]  H. V. Van Kruiningen,et al.  Ruminant paratuberculosis (Johne's disease): the current status and future prospects. , 1984, The Cornell veterinarian.

[11]  W. Fries,et al.  Quality of life in inflammatory bowel disease. , 1995, The Italian journal of gastroenterology.

[12]  L. Bermudez,et al.  Mycobacterium avium Grown inAcanthamoeba castellanii Is Protected from the Effects of Antimicrobials , 2000, Antimicrobial Agents and Chemotherapy.

[13]  R. Chiodini,et al.  Paratuberculosis: a potential zoonosis? , 1996, The Veterinary clinics of North America. Food animal practice.

[14]  J. Hermon-Taylor Mycobacterium avium subspecies paratuberculosis: the nature of the problem , 2001 .

[15]  I. Grant,et al.  Mycobacterium avium ssp. paratuberculosis and its potential survival tactics , 2006, Letters in applied microbiology.

[16]  A. E. Greenberg,et al.  Standard methods for the examination of water and wastewater : supplement to the sixteenth edition , 1988 .

[17]  O. Harb,et al.  Invasion of Protozoa by Legionella pneumophila and Its Role in Bacterial Ecology and Pathogenesis , 1998, Applied and Environmental Microbiology.

[18]  S Falkow,et al.  Interaction of Mycobacterium avium with environmental amoebae enhances virulence , 1997, Infection and immunity.

[19]  M. Collins,et al.  A model for analyzing growth kinetics of a slowly growing Mycobacterium sp , 1988, Applied and environmental microbiology.

[20]  N. Sung,et al.  Variation in Resistance of Mycobacterium paratuberculosis to Acid Environments as a Function of Culture Medium , 2003, Applied and Environmental Microbiology.

[21]  R. Whittington,et al.  Survival of Mycobacterium avium subsp. paratuberculosis in Dam Water and Sediment , 2005, Applied and Environmental Microbiology.

[22]  K. Birkness,et al.  Mycobacterium avium Bacilli Grow Saprozoically in Coculture with Acanthamoeba polyphaga and Survive within Cyst Walls , 1998, Applied and Environmental Microbiology.

[23]  Hugh Evans,et al.  Replication and Long-Term Persistence of Bovine and Human Strains of Mycobacterium avium subsp. paratuberculosis within Acanthamoeba polyphaga , 2006, Applied and Environmental Microbiology.