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Differential identification of Ascogregarina species (Apicomplexa: Lecudinidae) in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) by polymerase chain reaction.

We report 2 polymerase chain reaction (PCR)-based methods for distinguishing morphologically similar gregarine species based on amplification of variable regions of the internal transcribed spacer region of ribosomal DNA. The gregarines we investigated were Ascogregarina barretti (Vavra), A. culicis (Ross), and A. taiwanensis (Lien and Levine), parasites of the mosquitoes Ochlerotatus triseriatus (Say), Aedes aegypti (Linnaeus), and Ae. albopictus (Skuse), respectively. These 3 important vector mosquitoes often utilize the same container habitats, where larval development and infection by the parasite occurs, leaving ample opportunity for cross-species gregarine infection. Because previous studies have shown that the parasites A. culicis and A. taiwanensis variably affect fitness in both normal and abnormal mosquito hosts, distinguishing parasite infection and species is important. The task is complicated by the fact that these 2 parasite species are virtually identical in morphology, whereas A. barretti is morphologically distinct. Of the 2 PCR-based assays reported here, the first provides a rapid, sensitive, and straight-forward means of general ascogregarine detection based on a single PCR amplification. The second method provides a means of differentiation between A. culicis and A. taiwanensis based on a species-specific PCR assay. Together, these assays allow whole mosquitoes to be tested for the presence of Ascogregarina species as well as identification of both A. culicis and A. taiwanensis singly or in dual infections.

Aedes↗

Observations on associated histopathology with Aggregata octopiana infection (Protista: Apicomplexa) in Octopus vulgaris.

Gamogony and sporogony of Aggregata octopiana were commonly observed during histological examination of the digestive tract of wild Octopus vulgaris from Ria de Vigo (NW Spain). A. octopiana infected noncuticularized caecum and intestine, and cuticularized oesophagus and crop. Infection was also observed in the gills and in covering mesenterium, mainly of the digestive gland and gonad. Histological and ultrastructural lesions associated with A. octopiana included host cell hypertrophy with nuclear displacement, inflammation, phagocytosis, ulceration and destruction of organ architecture. The possible existence of a malabsorption syndrome in the host is deduced.

Animals↗

Malabsorption syndrome observed in the common octopus Octopus vulgaris infected with Aggregata octopiana (Protista: Apicomplexa).

Octopus vulgaris infected with Aggregata octopiana were collected from an open-water culture system in the Ría of Aldán (NW Spain). Digestive tract infection values were determined with the use of a Neubauer chamber by counting the number of A. octopiana sporocysts. After determining enzyme activity values by the colorimetric Api-Zym system Biomerieux, one representative enzyme of glycosidases, peptid hydrolases and phosphoric hydrolases showing high activity was spectrophotometrically analysed. The enzymes were maltase and leucine-aminopeptidase (LAP) involved in the absorption process, and acid phosphatase, a lysosomic enzyme, respectively. Enzymatic activity of maltase and LAP decreased significantly, with increased sporocyst counts. However, acid phosphatase activity increased with severity of infection, indicating the presence of degradative enzymes from phagocytic cells in the infected area. A detrimental effect on gastrointestinal function may result from a decrease or malfunction of absorption enzymes. The results suggest a malabsorption syndrome resulting from parasitic infection.

Acid Phosphatase↗

Hepatic Calyptospora sp. (Apicomplexa) infection in a wild-born, aquarium-held clutch of juvenile arapaima Arapaima gigas (Osteoglossidae).

From Manaus, Brazil, 12 juvenile arapaima Arapaima gigas were imported to the United States and sent to 2 public aquaria, 1 private hobbyist, and 1 retailer. All 12 fish became ill within 4 to 6 wk of arrival, with signs of anorexia, lethargy, depigmentation, and ascites, and subsequently died despite antibiotic and anthelminthic therapy. Gross necropsies of 7 fish revealed serosanguinous coelomic effusion in all 7 fish, and branchial monogeneosis in 3 of 6 fish. The monogeneans from 1 fish were identified as Dawestrema cycloancistrium (Ancyrocephalinae). Histologic examination of 7 fish showed a variety of lesions, principally in the liver, gills, brain and gastro-intestinal tract. Numerous coccidian oocysts replaced 15 to 33% of the liver parenchyma in 6 of 7 fish examined. Light and transmission electron microscopy revealed that each oocyst contained 4 pyriform sporocysts bearing numerous sporopodia on their tapering, posterior end; approximately 25 to 30% of the length of the sporocyst was adorned. Each sporocyst was covered by a thin, membranous veil, contained 2 sporozoites, and stained brilliant pink with the Ziehl-Neelsen acid-fast method. This morphology is consistent with that of Calyptospora sp. (Lack of fresh material precluded determination to species.) This is the first report of Calyptospora sp. in arapaima. The Calyptospora sp. infection probably contributed to the morbidity and mortality of the captive arapaima.

Animals↗

Hemosporid (Apicomplexa, Hematozoea, Hemosporida) community structure off pattern in wintering wild turkeys.

The hemosporid community of 76 wild turkeys (Meleagris gallopavo silvestris) from South Carolina (USA) was examined using thin blood smears collected during January and February 1994. High prevalences and low abundances of hemosporids characterized this community. Leucocytozoon smithi and Haemoproteus meleagridis occurred in 100% and 54% of the turkeys, respectively; a Plasmodium sp. was found in one bird. Prevalence of H. meleagridis was significantly higher in juvenile turkeys than adults, but prevalences did not differ significantly among four trap sites or by host sex. Mean (+/- SE) intensities of L. smithi, H. meleagridis, and Plasmodium sp. were 3.4 +/- 0.4, 1.8 +/- 0.3, and 3.0 per 10,000 erythrocytes, respectively. Abundances of L. smithi, H. meleagridis, and Plasmodium sp. were 3.4 +/- 0.9 +/- 0.2, and < 0.1 +/- < 0.1 per 10,000 erythrocytes, respectively. Juvenile turkeys had higher rank abundance values of L. smithi than adults, whereas no differences were found among trap sites or between sexes. No differences in rank abundances of H. meleagridis were found among trap sites, host age, or host sex variables. Collectively, both common hemosporid species varied by host age, reflecting higher abundances in juvenile turkeys. patterns of hemosporid prevalence appeared similar to patterns found in subtropical regions. Based on our data, we recommend using prevalence and abundance data to analyze the structure and pattern of hemosporid communities at the component community level.

Analysis of Variance↗

Amoebogregarina Nigra n. gen., n. comb. (Apicomplexa: Gregarinidae) from adult Melanoplus differentialis (Orthoptera: Acrididae) in southeastern Nebraska.

Amoebogregarina nigra n. gen., nov. comb. (Eugregarinida: Gregarinidae) is described from trophozoites, gamonts, associations, gametocysts, and oocysts collected from adult Melanoplus differentialis (Orthoptera: Acrididae) in Nemaha County, Nebraska. Host alimentary canals were examined for eugregarine parasites. Gregarines encountered were fixed as permanent specimens or subjected to a series of morphometric measurements. Morphometric analysis indicated the presence of Gregarina nigra, a poorly described taxon reported from a variety of Nearetic grasshopper species, Examination of G. nigra revealed a metamorphic epimerite assimilated by the protomerite on maturity. This epimerite-protomerite complex is unique within Gregarinidae, prompting creation of the genus Amoebogregarina. Amoebogregarina nigra is the type species in new combination. Gregarina indianensis is recognized as a junior synonym of A. nigra.

Animals↗

Neospora caninum (Apicomplexa) in a stillborn goat.

Tissue cysts of Neospora caninum were found in sections of brain from a stillborn pygmy goat. The tissue cysts had 1-2-microns-thick cyst walls and stained with anti-N. caninum serum in an immunohistochemical test. Glial nodules, mononuclear perivascular cuffing, and foci of inflammation were associated with N. caninum tissue cysts throughout the brain. This report indicates that N. caninum can be transmitted transplacentally in goats.

Animals↗

Prevalence of Hemolivia mauritanica (Apicomplexa: Adeleina: Haemogregarinidae) in natural populations of tortoises of the genus Testudo in the East Mediterranean region.

Hemolivia mauritanica (Sergent et Sergent, 1904) infections were found in 14% (n= 14) of Testudo graeca Linnaeus, 1758 tortoises in Bulgaria and in 92% (n = 26) of those in Turkey. Hemolivia mauritanica-like gametocytes were found in 81% (n = 47) of wild Testudo marginata Schoepff, 1792 tortoises in Greece. Parasitaemia intensity (the percentage of infected red blood cells found in approximately 104 cells) was 0.03-22.4% in T. graeca and 0.06-12.27% in T. marginata. On the other hand, all blood samples from 40 Bulgarian, 38 Greek, and 18 Croatian specimens of Testudo hermanni Gmelin, 1789 were negative.

Animals↗

Haemogregarine parasites (Apicomplexa: Hepatozoidae) in Caiman crocodilus yacare (Crocodilia: Alligatoridae) from Pantanal, Corumbá, MS, Brazil.

Haemogregarines were recorded in caimans Caiman crocodilus yacare from Pantanal. This study was carried out in seasonal ponds at the Miranda- Abobral subregion of Pantanal, State of Mato Grosso do Sul, western Brazil, from 1998 to 1999. Smears from 28 caimans were examined and 20 (71.4%) presented infection by a haemogregarine. Infections were observed in 11 males and 9 females. Morphological and morphometric observations suggest that the parasite forms found in this work are Hepatozoon caimani.

Alligators and Crocodiles↗

Sarcocystis neotomafelis sp. n (Protozoa; Apicomplexa) from the woodrat Neotoma micropus in Mexico.

The developmental cycle of a new species of Sarcocystis (S. neotomafelis) was investigated. Cysts obtained from the muscle fibers of the woodrats were given orally to 19 laboratory newborn kittens, 5 pups and 5 crotalids. Infection developed only in newborn kittens. Macrogametes, microgamonts and immature oocysts were observed principally in the jejunum. Immature oocysts were detected at 6-21 post infection days. Based on morphological observations made by light and electron microscopy, and repeating transmission experiments, S. neotomafelis is described as a new species. Also the prevalence of cysts and the statistical frequency of sarcocystosis according to location and hosts's sex are given. This is the first species described in Mexico, and also the first report for Neotoma.

Animals↗

[Clinical findings observed in rabbits, meat type, infected experimentally by Cystoisospora felis (Wenyon, 1923) Frenkel, 1977 (Apicomplexa: Cystoisosporinae)].

Twenty four rabbits type meat, cross-breed of White New Zealand and California, of both sexes, with age of 58 days old, and average weight of 1,586 g were used. Than, they were divided in three groups of eight rabbits each. The first group was inoculated orally, in two serial days, with 10(6) sporulated oocysts of Cystoisospora felis per animal, second group was considered per feed, and thirty group was left as control. Clinical signs as body weight, feed consumption, rectal temperature, and breathing frequency were taken daily. Carcasses weights were taking at 2, 4, 6, 8, 9, 16, 22 and 29 days after infection (DAI) when they were posted. In the first DAI, anorexia, slow movements, low percent of food consumption (85.76%), and of alimentary conversion were observed. Difference of 3.52% in the food consumption was observed between control and infected animals. The inoculated rabbits had high temperatures in the first DAI in comparison to controls and per fed animals (p<0.01) at 2nd and at 4th DAI. In relationship to the weight gain in the 1st week the carcasses of the infected rabbits had minimal difference in comparison to control animals. As conclusion, cystoisosporosis, beside clinical signs, can be interfered in the development of the rabbit.

Animals↗

Monocystis apporectodae sp. nov. (Protozoa: Apicomplexa: Eugregarinida), from an Indian earthworm Apporectodea trapezoides Duges.

A survey aimed at exploring the endoparasitic acephaline gregarine diversity in South-western Bengal, detected a new species of the genus Monocystis Stein, 1848, that resides in the seminal vesicles of the earthworm, Apporectodea trapezoides Duges collected in the district of Bankura from alluvial soil. Monocystis apporectodae sp. nov. is a ribbon-like organism with one or more prominent constric-tions especially in some mature forms and measures 178.0-224.0 (203.0+/-5.0) microm x 37.0-58.0 (46.0+/-1.5) microm. The extreme ends are pointed. Its gametocysts are ovoid and measure 108.0-118.0 microm (113.0+/-1.1) x 79.0-89.0 (83.0+/-1.1) microm. Oocysts are navicular in shape. The length of the oocysts ranges from 10.0-14.6 and the width, from 5.5-8.1 microm.

Animals↗

Monocystis metaphirae sp. nov. (Protista: Apicomplexa: Monocystidae) from the earthworm Metaphire houlleti (Perrier).

Biodiversity studies in search of endoparasitic acephaline gregarines revealed a new species of the genus Monocystis Stein, 1848 in the seminal vesicles of the earthworm Metaphire houlleti (Perrier) residing in alluvial soil of the district of North 24 Parganas. The new species is characterized by having bean-shaped gamonts measuring 94.0-151.0 (119.0+/-16.0) microm x 53.0-81.0(66.0+/-8.0) microm. The anterior end of the gamont is always wider than the posterior end. The mucron is always present at the wider end. The occurrence of syzygy (end to end, cauda-frontal) is a very rare feature which has been observed in the life cycle of the new species. The gametocyst is ovoid consisting of two unequal gamonts, measuring 85.0-102.0 microm (93.0+/-6.0). Oocysts are navicular in shape, measuring 6.5-11.0 (9.0+/-1.1) microm x 4.0-7.5 (5.5+/-1.9) microm.

Animals↗

A new species of Monocystis stein, 1848 (Protista: Apicomplexa: Eugregarinida) from the Indian earthworm, Amynthas Hawayanus Rosa, 1891 (Annelida: Oligochaeta).

As a part of an ongoing biodiversity survey of aseptate gregarine fauna of oligochaete hosts of West Bengal, an expedition was carried out in the Darjeeling district of West Bengal and most of the earthworms collected were found to be infested with a species of Monocystis Stein, 1848. The monocystid species was collected from the seminal vesicles of the earthworm and was identified as a new species, Monocystis amynthae sp. nov. The gamont of the new species is characterized by having an elongated body with broad anterior end, separated from the narrow posterior end by a prominent constriction measuring 49.0-77.0 (66.0+/-1.3) microm x 32.0-41.0 (37.0+/-2.8) microm. The gametocysts are oval-shaped, measuring 40.0-65.0 (58.0+/-2.1) microm. The oocysts are navicular, measuring 8.0-12.0 (10.5+/-1.1) microm x 4.0-6.0 (5.5+/-1.1) microm.

Animals↗

Goussia carpelli (Apicomplexa, Eimeriorina) from cyprinid fish: field observations and infection experiments.

Intestinal tissue samples of 9 cyprinid and gobiid fish species from several hatcheries and ponds in South Bohemia were found to harbour Goussia carpelli-like coccidian oocysts. Furthermore, laboratory transmission of Goussia carpelli, obtained from common carp, was attempted to 9 cyprinid fish species, both laboratory reared and obtained from pond environments. Infection experiments, carried out by fecal contamination (direct transmission) and by using tubificid oligochaetes as paratenic hosts resulted in weak infections in 4 out of 15 Carassius auratus used, but it was not possible to infect any of the other fishes used.

Animals↗