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Description of the Microsporidian Parasite, Heterosporis sutherlandae n. sp., Infecting Fish in the Great Lakes Region, USA.

Phelps NB, Mor SK, Armién AG, Pelican KM, Goyal SM - PLoS ONE (2015)

Bottom Line: A unique species of Heterosporis was identified, demonstrating less than 96% sequence identity to other published Heterosporis sp. on the basis of partial rRNA gene sequence analysis.Heterosporis sutherlandae n. sp. (formerly Heterosporis sp.) was identified in yellow perch (Perca flavescens), northern pike (Esox lucius) and walleye (Sander vitreus) from inland lakes in Minnesota and Wisconsin.Previous research suggests this species may be even more widespread in the Great Lakes region and should be reexamined using molecular techniques to better understand the distribution of this novel species.

View Article: PubMed Central - PubMed

Affiliation: Veterinary Diagnostic Laboratory, College of Veterinary Medicine, University of Minnesota, 1333 Gortner Avenue, Saint Paul, Minnesota, 55108, United States of America; Veterinary Population Medicine Department, College of Veterinary Medicine, University of Minnesota, 1365 Gortner Avenue, Saint Paul, Minnesota, 55108, United States of America.

ABSTRACT
Heterosporosis is an increasingly important microsporidian disease worldwide, impacting wild and farmed raised fishes in both marine and freshwater environments. A previously undescribed species (Heterosporis sp.), with widespread distribution in the Great Lakes region, was the subject of this study. Three angler-caught fish were submitted to the Minnesota Veterinary Diagnostic Laboratory from 2009-2010 with lesions caused by intracellular proliferation of parasitic spores, resulting in destruction and eventual widespread necrosis of the host skeletal muscles. Mature ovoid (5.8 x 3.5 μm) spores of a microsporidian parasite, consistent with the genus Heterosporis, were observed by light and electron microscopy. Molecular identification was performed using primer walking to obtain a near-complete rRNA gene sequence (~3,600 bp). A unique species of Heterosporis was identified, demonstrating less than 96% sequence identity to other published Heterosporis sp. on the basis of partial rRNA gene sequence analysis. Heterosporis sutherlandae n. sp. (formerly Heterosporis sp.) was identified in yellow perch (Perca flavescens), northern pike (Esox lucius) and walleye (Sander vitreus) from inland lakes in Minnesota and Wisconsin. Previous research suggests this species may be even more widespread in the Great Lakes region and should be reexamined using molecular techniques to better understand the distribution of this novel species.

No MeSH data available.


Related in: MedlinePlus

Heterosporis sutherlandae n. sp. from yellow perch and walleye.A) Sporophorocyst containing several sporophorous vesicles. The sporophorocyst is lined by a thick wall (double headed arrow). The inner surface of the sporophorocyst is indicated by arrows. The edge of the wall of one sporophorous vesicle is highlighted by arrowhead. Spore is marked by an asterisk. Scale bar is 2 μm. B) Magnification of the sporophorocyst wall (double headed arrow) in a muscle fiber (mf); note the thick external layer, which is formed of microtubules and filaments (tf) and an inner electron dense membrane (arrow). The most internal smooth electron dense layer is a sporophorous vesicle, which is formed of an electron dense amorphous material (white arrowhead). A sporoblast (sb) is near the wall of the sporophorous vesicle. Scale bar is 1 μm. C) Spore wall (sw) of 122–137 nm comprised of an electron dense exospore (ex) measuring 20.78–33.98 nm and an electron lucent endospore (en) measuring 101.76–103.36 nm. Scale bar is 200 nm. D) Longitudinal section of H. sutherlandae displaying a spore wall (wide arrow), posterior vacuole (Pv), coiled filament (white arrowhead), polar filament (white arrow), nucleus (N), and anterior polaroplast. Scale bar is 0.5 μm. Inset: Anchoring disk of a spore. Scale bar is 200 nm. E) Transverse section of the anterior pole of H. sutherlandae displaying a spore wall (wide arrow), polar filament (white arrow), and anterior (Pa) and posterior polaroplast (Pp). Scale bar is 200 nm. F) Longitudinal section of coiled filament of H. sutherlandae. Scale bar is 200 nm. G) Transverse section of polar filaments of H. sutherlandae showing a concentrically multilayer structure. Spore wall (sw). Scale bar is 200 nm.
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pone.0132027.g004: Heterosporis sutherlandae n. sp. from yellow perch and walleye.A) Sporophorocyst containing several sporophorous vesicles. The sporophorocyst is lined by a thick wall (double headed arrow). The inner surface of the sporophorocyst is indicated by arrows. The edge of the wall of one sporophorous vesicle is highlighted by arrowhead. Spore is marked by an asterisk. Scale bar is 2 μm. B) Magnification of the sporophorocyst wall (double headed arrow) in a muscle fiber (mf); note the thick external layer, which is formed of microtubules and filaments (tf) and an inner electron dense membrane (arrow). The most internal smooth electron dense layer is a sporophorous vesicle, which is formed of an electron dense amorphous material (white arrowhead). A sporoblast (sb) is near the wall of the sporophorous vesicle. Scale bar is 1 μm. C) Spore wall (sw) of 122–137 nm comprised of an electron dense exospore (ex) measuring 20.78–33.98 nm and an electron lucent endospore (en) measuring 101.76–103.36 nm. Scale bar is 200 nm. D) Longitudinal section of H. sutherlandae displaying a spore wall (wide arrow), posterior vacuole (Pv), coiled filament (white arrowhead), polar filament (white arrow), nucleus (N), and anterior polaroplast. Scale bar is 0.5 μm. Inset: Anchoring disk of a spore. Scale bar is 200 nm. E) Transverse section of the anterior pole of H. sutherlandae displaying a spore wall (wide arrow), polar filament (white arrow), and anterior (Pa) and posterior polaroplast (Pp). Scale bar is 200 nm. F) Longitudinal section of coiled filament of H. sutherlandae. Scale bar is 200 nm. G) Transverse section of polar filaments of H. sutherlandae showing a concentrically multilayer structure. Spore wall (sw). Scale bar is 200 nm.

Mentions: On toluidine blue stained plastic embedded sections, Heterosporis sp. was observed in the muscle tissue of the walleye and yellow perch. Spores were found free in the interstitium admixed in cell debris, within muscle fibers inside of sporophorocysts (Fig 2C) and within phagocytic vacuoles of macrophages (Fig 3A and 3B). Large numbers of spores within macrophages were in different stages of degradation (Fig 3A and 3B). A few spores were present in the subsarcolema (Fig 3A). Sporophorocysts containing all developmental stages within sporophorous vesicles were observed in both walleye and yellow perch tissues (Fig 4A). However, very few sporophorous vesicles containing meronts stage were observed. The sporophorocystic wall derived presumably from the host cell was compounded with one thick layer of different electron densities. The outermost interface was composed of packed intermediary filaments, microtubules and cisternae from the endoplasmic reticulum (Fig 4A and 4B). Lining the innermost surface of the sporophorocystic wall was an electron dense smooth layer (Fig 4B). A sporophorocyst contained a few to large numbers of sporophorous vesicles, which had an average of 4–8 maturating spores with a maximum of 13 (Table 2). These were embedded in an electron opaque matrix.


Description of the Microsporidian Parasite, Heterosporis sutherlandae n. sp., Infecting Fish in the Great Lakes Region, USA.

Phelps NB, Mor SK, Armién AG, Pelican KM, Goyal SM - PLoS ONE (2015)

Heterosporis sutherlandae n. sp. from yellow perch and walleye.A) Sporophorocyst containing several sporophorous vesicles. The sporophorocyst is lined by a thick wall (double headed arrow). The inner surface of the sporophorocyst is indicated by arrows. The edge of the wall of one sporophorous vesicle is highlighted by arrowhead. Spore is marked by an asterisk. Scale bar is 2 μm. B) Magnification of the sporophorocyst wall (double headed arrow) in a muscle fiber (mf); note the thick external layer, which is formed of microtubules and filaments (tf) and an inner electron dense membrane (arrow). The most internal smooth electron dense layer is a sporophorous vesicle, which is formed of an electron dense amorphous material (white arrowhead). A sporoblast (sb) is near the wall of the sporophorous vesicle. Scale bar is 1 μm. C) Spore wall (sw) of 122–137 nm comprised of an electron dense exospore (ex) measuring 20.78–33.98 nm and an electron lucent endospore (en) measuring 101.76–103.36 nm. Scale bar is 200 nm. D) Longitudinal section of H. sutherlandae displaying a spore wall (wide arrow), posterior vacuole (Pv), coiled filament (white arrowhead), polar filament (white arrow), nucleus (N), and anterior polaroplast. Scale bar is 0.5 μm. Inset: Anchoring disk of a spore. Scale bar is 200 nm. E) Transverse section of the anterior pole of H. sutherlandae displaying a spore wall (wide arrow), polar filament (white arrow), and anterior (Pa) and posterior polaroplast (Pp). Scale bar is 200 nm. F) Longitudinal section of coiled filament of H. sutherlandae. Scale bar is 200 nm. G) Transverse section of polar filaments of H. sutherlandae showing a concentrically multilayer structure. Spore wall (sw). Scale bar is 200 nm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4526549&req=5

pone.0132027.g004: Heterosporis sutherlandae n. sp. from yellow perch and walleye.A) Sporophorocyst containing several sporophorous vesicles. The sporophorocyst is lined by a thick wall (double headed arrow). The inner surface of the sporophorocyst is indicated by arrows. The edge of the wall of one sporophorous vesicle is highlighted by arrowhead. Spore is marked by an asterisk. Scale bar is 2 μm. B) Magnification of the sporophorocyst wall (double headed arrow) in a muscle fiber (mf); note the thick external layer, which is formed of microtubules and filaments (tf) and an inner electron dense membrane (arrow). The most internal smooth electron dense layer is a sporophorous vesicle, which is formed of an electron dense amorphous material (white arrowhead). A sporoblast (sb) is near the wall of the sporophorous vesicle. Scale bar is 1 μm. C) Spore wall (sw) of 122–137 nm comprised of an electron dense exospore (ex) measuring 20.78–33.98 nm and an electron lucent endospore (en) measuring 101.76–103.36 nm. Scale bar is 200 nm. D) Longitudinal section of H. sutherlandae displaying a spore wall (wide arrow), posterior vacuole (Pv), coiled filament (white arrowhead), polar filament (white arrow), nucleus (N), and anterior polaroplast. Scale bar is 0.5 μm. Inset: Anchoring disk of a spore. Scale bar is 200 nm. E) Transverse section of the anterior pole of H. sutherlandae displaying a spore wall (wide arrow), polar filament (white arrow), and anterior (Pa) and posterior polaroplast (Pp). Scale bar is 200 nm. F) Longitudinal section of coiled filament of H. sutherlandae. Scale bar is 200 nm. G) Transverse section of polar filaments of H. sutherlandae showing a concentrically multilayer structure. Spore wall (sw). Scale bar is 200 nm.
Mentions: On toluidine blue stained plastic embedded sections, Heterosporis sp. was observed in the muscle tissue of the walleye and yellow perch. Spores were found free in the interstitium admixed in cell debris, within muscle fibers inside of sporophorocysts (Fig 2C) and within phagocytic vacuoles of macrophages (Fig 3A and 3B). Large numbers of spores within macrophages were in different stages of degradation (Fig 3A and 3B). A few spores were present in the subsarcolema (Fig 3A). Sporophorocysts containing all developmental stages within sporophorous vesicles were observed in both walleye and yellow perch tissues (Fig 4A). However, very few sporophorous vesicles containing meronts stage were observed. The sporophorocystic wall derived presumably from the host cell was compounded with one thick layer of different electron densities. The outermost interface was composed of packed intermediary filaments, microtubules and cisternae from the endoplasmic reticulum (Fig 4A and 4B). Lining the innermost surface of the sporophorocystic wall was an electron dense smooth layer (Fig 4B). A sporophorocyst contained a few to large numbers of sporophorous vesicles, which had an average of 4–8 maturating spores with a maximum of 13 (Table 2). These were embedded in an electron opaque matrix.

Bottom Line: A unique species of Heterosporis was identified, demonstrating less than 96% sequence identity to other published Heterosporis sp. on the basis of partial rRNA gene sequence analysis.Heterosporis sutherlandae n. sp. (formerly Heterosporis sp.) was identified in yellow perch (Perca flavescens), northern pike (Esox lucius) and walleye (Sander vitreus) from inland lakes in Minnesota and Wisconsin.Previous research suggests this species may be even more widespread in the Great Lakes region and should be reexamined using molecular techniques to better understand the distribution of this novel species.

View Article: PubMed Central - PubMed

Affiliation: Veterinary Diagnostic Laboratory, College of Veterinary Medicine, University of Minnesota, 1333 Gortner Avenue, Saint Paul, Minnesota, 55108, United States of America; Veterinary Population Medicine Department, College of Veterinary Medicine, University of Minnesota, 1365 Gortner Avenue, Saint Paul, Minnesota, 55108, United States of America.

ABSTRACT
Heterosporosis is an increasingly important microsporidian disease worldwide, impacting wild and farmed raised fishes in both marine and freshwater environments. A previously undescribed species (Heterosporis sp.), with widespread distribution in the Great Lakes region, was the subject of this study. Three angler-caught fish were submitted to the Minnesota Veterinary Diagnostic Laboratory from 2009-2010 with lesions caused by intracellular proliferation of parasitic spores, resulting in destruction and eventual widespread necrosis of the host skeletal muscles. Mature ovoid (5.8 x 3.5 μm) spores of a microsporidian parasite, consistent with the genus Heterosporis, were observed by light and electron microscopy. Molecular identification was performed using primer walking to obtain a near-complete rRNA gene sequence (~3,600 bp). A unique species of Heterosporis was identified, demonstrating less than 96% sequence identity to other published Heterosporis sp. on the basis of partial rRNA gene sequence analysis. Heterosporis sutherlandae n. sp. (formerly Heterosporis sp.) was identified in yellow perch (Perca flavescens), northern pike (Esox lucius) and walleye (Sander vitreus) from inland lakes in Minnesota and Wisconsin. Previous research suggests this species may be even more widespread in the Great Lakes region and should be reexamined using molecular techniques to better understand the distribution of this novel species.

No MeSH data available.


Related in: MedlinePlus