Three new fleas from Sulawesi, Indonesia (Siphonaptera: Pygiopsyllidae & Ceratophyllidae)

Gryphopsylla maxomydis n. sp. (Pygiopsyllidae), Medwayella rubrisciurae n. sp. (Pygiopsyllidae) and Macrostylophora theresae n. sp. (Ceratophyllidae) are described from endemic rodents in Sulawesi. Gryphopsylla maxomydis was collected from the murids Maxomys musschenbroekii and Paruromys dominator in Central Sulawesi (Sulawesi Tengah). However, M. musschenbroekii appears to be the true host of this flea because it has spiny pelage and G. maxomydis shows morphological adaptations for parasitizing spiny hosts including a remarkable "beak-like" structure on the head. This adatation is similar to a beak-like structure on the head of Gryphopsyllo hopkinsi (Traub) which parasitizes the spiny murid Maxomys whiteheadi in Borneo (Sabah). Medwayella rubrisciurae was collected from the large tree squirrel Rubrisciurus rubriventer in Central Sulawesi and this represents the first report of this flea genus in Sulawesi. Macrostylophora theresce was recorded from the murids Bunomys fratrorum, P. dominator and Rattus xanthurus in North Sulawesi (Sulawesi Utara); most other members of this flea genus parasitize squirrels in the Oriental and Palaearctic zoogeographical regions.     

Two new species of Stivalius (Siphonaptera--Pygiopsyllidae--Pygiopsyllinae) from Indonesia

The fleas, S. meridionalis n. sp. from Timor and S. franciscae n. sp. from Sulawesi, Indonesia are described and illustrated. S. meridionalis was collected in Central Timor (Timor Tengah) at elevations between 930 m and 1,520 m, mainly from the murine rodent Rattus tanezumi, although one specimen was recovered from a house shrew, Suncus murinus. S. franciscae was collected in Central Sulawesi (Sulawesi Tengah) at known elevations between 950 m and 2,255 m from six species of native endemic murine rodents, especially Rattus hoffmanni, and from two species of commensal murine rodents. One specimen of S. franciscae was also collected from the native endemic murine Paruromys dominator in North Sulawesi (Sulawesi Utara) at 1,780 m. These two new flea species are compared with the previously described taxa included in the genus Stivalius sensu stricto.     

Gymnomeropsylla n. gen. (Siphonaptera: Pygiopsyllidae) from Sulawesi,Indonesia, with the description of two new species

Compared to related genera, this new flea genus is characterized by the absence, or presence of very few, bristles on the external surface of femur I and especially by the morphology of the apex of sternite IX in the male, which is hyaline and lacks spiniform bristles. The two new species, G. bunomydis and G. margaretamydis, are distinguished from each other by the structure of the genitalia, and the presence of numerous erect bristles on the thorax and abdominal tergites of the latter species. Both of these new species parasitize murine rodents that are endemic to Sulawesi; G. bunomydis was collected mainly from Bunomys chrysocomus and G. margaretamydis only from Margaretamys parvus.     

Morphological and biochemical correlates in the characterization of Sarcocystis spp

Isoenzyme electrophoretic techniques were applied to the characterization of seven Sarcocystis spp. that had been identified by conventional morphological studies. Cystozoites were harvested from macroscopic cysts from sheep, cattle, and mice and from microscopic cysts from sheep, cattle, and goats. Soluble cystozoite extracts were subjected to cellulose acetate gel electrophoresis and characterized at 15 of the 39 enzyme loci examined. Genetic relationships among isolates were examined by simple phenetic clustering. Two different morphological types of macroscopic cysts from sheep, identified as S. gigantea (syn. S. ovifelis) and S. medusiformis, consistently differed at 40% of the loci examined. Such genetic divergence confirms their separate morphotypic classification. Both differed from microscopic cyst isolates from sheep at 87% of the loci examined; however, two different morphotypes of microscopic cysts were found in the sheep sampled (thick-walled and thin-walled cysts). Until sufficient numbers of each type can be isolated and examined separately, both were regarded as belonging to the species S. tenella (syn. S. ovicanis). Macroscopic and microscopic cysts from cattle consistently differed at 80% of the loci thereby supporting their separate classification as S. hirsuta (syn. S. bovifelis) and S. cruzi (syn. S. bovicanis), respectively. Isolates from goats (microscopic cysts identified as S. capracanis) differed from S. tenella and S. cruzi at 20% and 47% of the loci, respectively. All macroscopic cyst isolates from the various host animal species (including S. muris from mice) differed from each other at nearly all loci. Isoenzyme electrophoretic techniques therefore provided genetic evidence supporting the classification of these various Sarcocystis spp. by their morphological characteristics.     

Ultrastructure of the cysts of Sarcocystis grueneri from cardiac muscle of reindeer (Rangifer tarandus tarandus)

Cysts of Sarcocystis grueneri from cardiac muscle of reindeer (Rangifer tarandus) in Norway were examined by transmission electron microscopy. The limiting unit membrane of the cyst proper formed regularly spaced invaginations into the cyst at numerous sites coinciding with interruptions in the underlying osmiophilic layer. The primary cyst wall formed numerous strip-like, sinuous protrusions, which were 30-40 nm thick, 150-300 nm wide and up to 4.5 microns long, and were running in parallel with the surface of the cyst. Generally the protrusions were arranged in several closely spaced layers compressed against the cyst. The nature and arrangement of the protrusions render them undetectable by light microscopy. Cyst ground substance divided the interior of the cyst into compartments containing typical sarcosporidian metrocytes and cystozoites. The cysts of S. grueneri from reindeer were ultrastructurally similar to cysts reported from red deer, roe deer and moose by other workers. The possibility that these cervids are hosts for a common Sarcocystis species is discussed.     

Morphological and Biochemical Correlates in the Characterization of Sarcocystis spp.1

ABSTRACT. Isoenzyme electrophoretic techniques were applied to the characterization of seven Sarcocystis spp. that had been identified by conventional morphological studies. Cystozoites were harvested from macroscopic cysts from sheep, cattle, and mice and from microscopic cysts from sheep, cattle, and goats. Soluble cystozoite extracts were subjected to cellulose acetate gel electrophoresis and characterized at 15 of the 39 enzyme loci examined. Genetic relationships among isolates were examined by simple phenetic clustering. Two different morphological types of macroscopic cysts from sheep, identified as S. gigantea (syn. S. ovifelis) and S. medusiformis, consistently differed at 40% of the loci examined. Such genetic divergence confirms their separate morphotypic classification. Both differed from microscopic cyst isolates from sheep at 87% of the loci examined; however, two different morphotypes of microscopic cysts were found in the sheep sampled (thick-walled and thin-walled cysts). Until sufficient numbers of each type can be isolated and examined separately, both were regarded as belonging to the species S. tenella (syn. S. ovicanis). Macroscopic and microscopic cysts from cattle consistently differed at 80% of the loci thereby supporting their separate classification as S. hirsuta (syn. S. bovifelis) and S. cruzi (syn. S. bovicanis), respectively. Isolates from goats (microscopic cysts identified as S. capracanis) differed from S. tenella and S. cruzi at 20% and 47% of the loci, respectively. All macroscopic cyst isolates from the various host animal species (including S. muris from mice) differed from each other at nearly all loci. Isoenzyme electrophoretic techniques therefore provided genetic evidence supporting the classification of these various Sarcocystis spp. by their morphological characteristics.     

Ultrastructure of the cyst wall of Sarcocystis spp. from some rodents in Malaysia.

The cyst wall of Sarcocystis cysts from the skeletal muscles and subcutaneous tissues of 4 species of rats and 1 species of bandicoot in Malaysia was examined under the electron microscope. Three types of sarcosysts with morphologically distinct cyst walls were found in these rodents. These morphologically distinct types of sarcocysts occurred as single or mixed infections in their various rodent hosts, suggesting that these rodents are important, though non-specific, intermediate hosts in the life cycle of at least 3 species of Sarcocystis. The final hosts of these Sarcocystis species are unknown.     

The flea genus Sigmactenus (Siphonaptera: Leptopsyllidae): three new taxa from Sulawesi, updated identification key, and distribution map for all known species and subspecies

One new species and two new subspecies of fleas are described. These are S. sulawesiensis n. sp. from North and Central Sulawesi, S. alticola pilosus n. ssp. from Central Sulawesi, and S. alticola crassinavis n. ssp. from North Sulawesi. All three of these new taxa are ectoparasites of native, endemic murine rodents. Two of the new taxa, S. sulawesiensis and S. alticola crassinavis, coexist on the same mountain, Gunung Moajat, in North Sulawesi. The related S. alticola alticola, which becomes the nominate subspecies, parasitises the murine rodent Maxomys alticola in northern Borneo (Sabah) and it is hypothesized that Sigmactenus first colonized Sulawesi as an ectoparasite of ancestral Maxomys, or perhaps Rattus, as these murines dispersed from southeast Asia to Sulawesi; 15 endemic murine rodent species belonging to these two genera are known to currently inhabit Sulawesi. An identification key and distribution map are included for all known species and subspecies of Sigmactenus. In addition to the three new taxa and S. a. alticola, these include: S. celebensis from South Sulawesi, S. timorensis from Timor, S. toxopeusi from New Guinea, and S. werneri from the Philippines (Mindanao and Negros).     

First report of fossilized cysts produced by the benthic Bysmatrum subsalsum (Dinophyceae) from a shallow Mexican lagoon in the Gulf of Mexico

Cysts belonging to the benthic dinoflagellate Bysmatrum subsalsum were recovered from palynologically treated sediments collected in the Alvarado Lagoon (southwestern Gulf of Mexico). The cysts are proximate, reflecting the features of the parent thecal stage, and their autofluorescence implies a dinosporin composition similar to the cyst walls of phototrophic species. This finding is important for our understanding of B. subsalsum life cycle transitions and ecology. Encystment may play an important role in the bloom dynamics of this species as it can enable the formation of a sediment cyst bank that allows reinoculation of the water column when conditions become favorable. This is the first report of a fossilized cyst produced by a benthic dinoflagellate recovered from sub‐recent sediments.     

Morphological studies on the muscle cysts of Sarcocystis dirumpens (Hoare 1933) H?fner and Matuschka 1984 in several host species revealing endopolygeny in metrocytes

Muscle cysts from rodents (Mastomys natalensis, Mus musculus, Rattus norvegicus, Meriones unguiculatus, Phodopus sungorus) experimentally infected with Sarcocystis dirumpens were examined by light and electron microscopy. The corrugated primary wall showed a pattern of densely arranged invaginations surrounded by minute bleb-like evaginations. True protrusions were absent. The length of the blebs and the thickness of the primary wall varied insignificantly but not remarkable host-dependent morphological differences could be noticed between the cyst wall structures 60-418 days p.i. In the mature parts of the cysts merozoites and metrocytes showed typical apicomplexan features of fine structure and mode of multiplication (endodyogeny). In the tips of the cysts large metrocytes simultaneously formed more than two daughter cells (at least up to 12) by endopolygeny.     

A comparison of Giardia microti and Spironucleus muris cysts in the vole: an immunocytochemical, light, and electron microscopic study

We have shown that cysts of the genus Spironucleus share many common morphological features with Giardia cysts including: 2-4 nuclei, flagellar axonemes, a distinct cyst wall, and they even display the same immunostaining as Giardia cysts when labeled with antibodies specific for Giardia cyst wall. A direct comparison of Spironucleus muris and Giardia microti cysts have revealed that cysts of S. muris are significantly smaller than cysts of G. miroti. At the ultrastructural level, the cyst walls are similar in fibrillar appearance, but the width of the S. muris cyst wall is significantly less than that of G. microti. The cysts of S. muris also differ from G. microti in that they contain a striated rootlet fiber, flagellar sheath, and numerous glycogen rosettes. Characteristic features of Giardia include the adhesive disc and median body. Although the cysts of Spironucleus and Giardia are similar in appearance, these unique morphological features can be used to distinguish between the 2 protozoa and should be employed in the detection of Giardia cysts in water samples.     

New species of the fleas Farhangia and Nestivalius,from endemic rodents in Sulawesi,Indonesia

The fleas (Siphonaptera: Pygiopsyllidae) Farhangia quattuordecimdentata sp. n. and Farhangia sedecimdentata sp. n. are described from pygmy squirrels (Prosciurillus spp.), and Nestivalius sulawesiensis sp. n. from murine rodents, all from Sulawesi, Indonesia. Both new species of Farhangia were collected in Central Sulawesi (Sulawesi Tengah); F. quattuordecimdentata sp. n. was recovered mainly from P. murinus, whereas F. sedecimdentata sp. n. was recovered mainly from P. leucomus. These new species are compared with the two previously described species of Farhangia: F. celebensis (Ewing) from P. murinus in North Sulawesi (Sulawesi Utara) and F. sciuri (Ewing) from the tree squirrel Callosciurus prevosti in Sabah (Borneo). Nestivalius sulawesiensis sp. n. was collected from six species of endemic murine rodents in both North and Central Sulawesi. It is compared to the morphologically similar N. pomerantzi (Traub) from Mindanao, which parasitizes murine hosts that are endemic to the Philippines.     

A Light Microscopic Comparison of The Cysts of Four Species of Sarcocystis Infecting The Domestic Reindeer (Rangifer Tarandus) in Northern Norway

Fresh preparations of micro-isolated sarcocysts from skeletal and cardiac muscle of 12 reindeer were examined by light microscopy. On the basis of cyst structure and cyst wall structure 4 Sarcocystis spp. could be differentiated. New names have been proposed for 2 previously unnamed Sarcocystis spp. of reindeer, and S. grueneri has been redefined. S. rangiferi n. sp. had macroscopic cysts in skeletal muscle measuring 2106×403 µm. The cyst wall protrusions were finger-like and measured 13.2×6.7 µm. The cysts were surrounded by a layer of fibrillar material. S. tarandi n. sp. had micro- to macroscopic cysts primarily in skeletal muscle, but a few cysts were found in the heart of one animal. In skeletal muscle the cysts measured 999×75µm; in the heart the cysts were shorter and wider. The cyst wall protrusions were fingerlike and measured 9.2×2.2 µm. S. grueneri had micro- to macroscopic cysts in cardiac muscle measuring 581×137 µm. The cyst wall was thin and relatively smooth with no visible protrusions. Sarcocystis sp. had micro- to macroscopic, slender cysts in skeletal muscle measuring 916×64 µm. The cyst wall had tightly packed, short, knob-like protrusions. The cysts of this species were previously classified as cysts of S. grueneri.     

Ultrastructure of the cyst wall of sarcocysts of the roe deer (Capreolus capreolus L.) (author's transl)]

The ultrastructure of the cyst wall of two types of sarcocysts from roe deer is described. In the thin-walled cyst (wall thickness 0.18-00.26 micrometer), the primary cyst wall forms long, finger-shaped protrusions distant from one another and running in parallel with the surface of the cyst (Figs. 1a--d, FP). No fibrils are observable in the protrusions. The primary cyst wall between them forms numerous bubble-like invaginations (Fig. 1c, arrows). In the thick-walled cysts (wall thickness 4.9--7.49 micrometer), the primary cyst wall forms massive, palisade-like protrusions lying close one to another (Figs. 2a, c). There are numerous fibrillar and tubular structures in these protrusions (Fig. 2d), and the primary cyst wall occasionally forms shallow invaginations at the base of some protrusions (Fig. 2b). The unit membrane onthe surface of some protrusions is slightly undulated and covered with a layer of short and thick bars on the outside. The sarcocysts found in roe deer are compared with those from cattle and sheep.     

Effects of frozen storage on the structure of sarcocysts in pig muscle and implications in taxonomic studies

The morphology of the cyst wall of Sarcocystis has unique characteristics that can be used in species identification. To find a suitable way to preserve Sarcocystis cyst samples for species identification, by light microscopy and electron microscopy, we recorded the morphological changes in the cysts of Sarcocystis suihominis and Sarcocystis miescheriana from pig muscle, induced by storage at -20 degrees C. Comparisons were made between fresh cysts and those subjected to frozen storage for periods of 3 days, 20 days and 30 days. Results: cyst wall of the two Sarcocystis species appeared unaffected by storage. There was no obvious change in the length, nor in the width of the protrusions after storage (P>0.05), but the structure of the bradyzoite in the sarcocyst was in many cases disintegrated at -20 degrees C in 20 days for S. miescheriana and 30 days for S. suihominis. To our knowledge this is the first report that Sarcocystis cyst in muscle can be stored at -20 degrees C before and remain suitable for ultrastructural morphological study. Consequently, this paper proposes freezing as a convenient storage method for samples used in taxonomic studies of Sarcocystis.     

Cytochemical study of the cysts of the sarcosporidian Sarcocystis bovicanis. I. Nucleic acids, polysaccharides, lipids and proteins

A light microscopic study of S. bovicanis cysts and cyst stages has been carried out, in addition to morphological characterization of cysts. At least two types of cyst stages could be distinguished--merozoites and metrocytes. The light microscopic differentiation of the third type--the intermediate cells--from merozoites seems to be rather difficult especially when non-dividing cells are examined. Merozoites (zoites) much varied in size, and besides the usual parasitic cells with the terminal nuclei, cells with the central ones were recognized. Since the classical Feulgen reaction did not give sufficient results when establishing DNA distribution, its modification with a fluorescent agent Auramin O was used. The latter provided excellent results showing numerous chromatin granules in the nucleus, no distinct nucleoli being determined. Gallocyanin--chromalum method and methyl green--pyronin staining for DNA and RNA demonstrated a poor staining of the nucleus contrasting with an intensive coloration of cytoplasmic RNA and associated high level protein synthesis. The PAS reaction revealed numerous polysaccharide granules in the cytoplasm of zoites. On cryostat sections a certain PAS positive layer was distinguished around the cyst in the muscle tissue which did not disappear even after a long term amylase treatment. Even more intensively stained was the pre-cystic muscle after cytochemical test for general protein using amido black and coomassie blue. It does not seem unlikely that some metabolic changes may occur in the host cell harbouring the cyst. Several methods for lipid detection in cyst stages with Fat red, Oil red O and Sudan black B were used with negative results.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fine structure and development of Sarcocystis aucheniae in llamas

Isolated cysts of Sarcocystis aucheniae of the llama (Lama glama) were fed to one dog and one cat. Only the dog excreted sporocysts, measuring 13.1-15.7 (15.0 +/- 0.54) X 9.0-11.3 (10.4 +/- 0.36) micron after 11 days for 21 days. A second cat, which had ingested meat of a llama containing macrocysts of S. aucheniae as well as sarcosporidial cysts visible only under a microscope also did not excrete sporocysts. The cysts of S. aucheniae are surrounded by a folded primary cyst wall forming cauliflower-like protrusions into the muscle fibre. The protrusions contain numerous microfilaments. In addition, the primary cyst wall forms numerous tiny vesicles. The parasitized muscle fibre is located in a large cavity within the normal muscle tissue. The cyst wall of S. aucheniae is similarly structured to that of S. gigantea of the sheep.     

Ultrastructural development of the sarcocyst of Sarcocystis rauschorum (Apicomplexa: Sarcocystidae) in the varying lemming Dicrostonyx richardsoni

The development of the sarcocyst of Sarcocystis rauschorum in its intermediate host was studied. Lemmings were orally administered sporocysts of S. rauschorum obtained from snowy owls (Nyctea scandiaca). Beginning at 9 days postinoculation (DPI) and at various intervals to 84 DPI, skeletal muscle tissue taken from the infected lemmings was examined by electron microscopy. At 9 DPI the sarcocysts contained few metrocytes and the cyst wall was flat. The metrocytes underwent endodyogeny, and within a few days the cyst wall of the rapidly growing sarcocyst developed numerous tubulovesicular invaginations into the electron-dense layer, and the wall had a few irregular infoldings. By 21 DPI, banana-shaped bradyzoites appeared, and by 84 DPI the mature cysts were filled with bradyzoites in groups subdivided by septa and by deep infoldings of the cyst wall. The fine structure of the wall remained simple throughout maturation, with no conspicuous invagination or protrusion. The sarcocyst produced in response to S. rauschorum is unlike those from many species of Sarcocystis, which have complex walls that change markedly as the sarcocysts mature; however, its simple appearance is similar to other species that have rodents as intermediate hosts and raptorial birds as definitive hosts.     

Myositis and death in bobwhites, Colinus virginianus (L.), due to hemorrhagic cysts of a haemosporozoan of undetermined taxonomic status

An epizootic of myositis and death in pen-reared bobwhites occurred at a hunting club in California. The myositis was caused by myriads of elongate protozoan cysts. The cysts were in various stages of development and when mature, contained spherical zoites 1 micron in diameter. Sinuous compartments were present in all cysts. The walls of the compartments were composed of a material similar to the cyst walls. Mature cysts were filled with blood. The histologic and ultrastructural morphology of the parasite revealed the parasite to be a haemosporozoan. The parasite has numerous similarities to both Akiba caulleryi (Mathis and Leger, 1909) and organisms that cause aberrant leucocytozoonosis in other species of birds. Further studies are needed to determine if the parasite is a part of the normal parasite fauna of quail or if it represents a parasite in an aberrant host.