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Fine structure of the oocyst walls of Isospora serini and Isospora canaria and excystation of Isospora serini from the canary, Serinus canarius L.

Oocysts of Isospora serini and Isospora canaria, from the canary Serinus canarius, were broken, added to a cell suspension, fixed in Karnovsky's fluid, and studied in the electron microscope. The oocyst wall of each species had an electron-lucent inner layer, a more osmiophilic middle layer and an outer layer of electron-lucent (I. serini) or electron-dense material interspersed with some electron-lucent material (I. canaria). A few, relatively large lipid-like bodies were present in the outer or middle layer of the oocyst wall of I. canaria. As many as 9 membranes were present in the oocyst wall of I. canaria and 3 in that of I. serini. When exposed to a trypsin-sodium taurocholate fluid, sporozoites of I. serini excysted from 5-month-old sporocysts in vitro, but not from sporocysts stored for more than 6 months. No excystation occurred in 15-month-old I. canaria sporocysts. Similarities and differences in excystation between I. serini and other Isospora, Eimeria, and Sarcocystis species are discussed.

Animals

Excystation of Isospora arctopitheci Rodhain, 1933 with notes on a similiar process in Isospora begemina (Stiles, 1891) Lühe, 1906.

The in vitro excystation process of sporozoites of Isospora arctophitheci Rodhain, 1933 from the titi marmoset Saguinus geoffroyi and of Isospora bigemina (Stiles, 1891) Lühe, 1906 from the bobcat, Lynx rufus is presented. Sporocysts of both species lack a Stieda body and when exposed to a trypsin-sodium taurocholate (pH 7.4) excysting fluid the walls of both collapse in a similar fashion, along apparently predetermined lines. Similarities and differences on excystation between. I. arctopitheci, I. bigemina, and other Isospora, Eimeria, and Sarcocytis species are summarized. Such studies show that 2 distinct patterns of sporozoite excystation have been described to date, and both appear to be related to the structure of the sporocyst.

Animals

Exogenous stages of Isospora serini (Aragão)and Isospora canaria sp. n. in the canary (Serinus canarius Linnaeus).

Exogenous stages of Isospora serini (Aragao) and Isospora canaria sp. n. from the canary (Serinus canarius Linnaeus) are described. Oocytes of I. serini are spheroid and average 19.2 times 20.1 mum, while those of I. canaria are larger, more ellipsoid, and average 21.8 times 24.6 mum. No oocyst residuum is present and the oocyst walls of both species are colorless, transparent, and single layered. Sporocysts average 9.4 times 15.2 mum for I. serini and 11.5 times 18.1 mum for I. canaria. The I. canaria sporocyst has a substiedal body, but none was found in I. serini sporocysts. Both species have a spherical sporocyst residuum; this was obscured in the I. serini sporocyst by scattered granules. Living sporozoites of I. canaria average 3.6 times 16.9 mum and have 1 to 3 refractile globules; those of I. serini have 2 globules and average 2.8 times 12.6 mum. A disseminated infection of the mononuclear phagocytes results from administration of I. serini while I. canaria oocysts give rise to a typical coccidian infection restricted to the intestinal epithelium. Asexual stages of I. serini in macrophages are indistinguishable from parasites previously called avian Toxoplasma, Atoxoplasma, and Lankesterella.

Animals

Isospora mcquistioni and Isospora bioccai (Apicomplexa, Eimeriidae): two new coccidian parasites from Carduelis sinica (Passeriformes, Fringillidae).

The following two species are described from Carduelis sinica (Greenfinch) from Italy. The oocysts of Isospora mcquistioni n. sp. were 26.0 x 22.6 (24.0-28.5 x 20.0-24.2) microns and ovoid with a smooth bilayered wall. Neither micropyle nor oocyst residuum were observed. One polar granule was found. Sporocysts were oval, 18.1 x 11.4 (16.0-19.8 x 11.0-12.0) microns, and with a symmetrical Stieda complex. The residuum was compact and spherical. Isospora bioccai n. sp. oocysts were spherical to subspherical and 24.0 x 23.6 (22.0-26.0 x 21.0-25.8) microns. The oocyst wall was smooth and bilayered. A micropyle and oocyst residuum were absent; 4 to 10 elongate polar granules were present. Sporocysts were 19.5 x 11.6 (18.0-20.0 x 10.0-12.4) microns, ellipsoidal, and with a symmetrical Stieda complex. The sporocyst residuum was diffuse and composed of a few granules.

Animals

Isospora belli infection. A report of two cases in patients with AIDS.

OBJECTIVE: To present the findings of Isospora belli infection in two patients with the acquired immunodeficiency syndrome (AIDS). CLINICAL FEATURES: One patient was part Aboriginal, the other an immigrant recently arrived from Peru. Both men were infected with the human immunodeficiency virus (HIV). They presented with watery diarrhoea, one with severe weight loss. Isospora belli oocysts were detected in multiple faecal specimens from both patients. One patient had a concomitant infection with Cryptosporidium sp. INTERVENTION AND OUTCOME: There was a rapid response to treatment with cotrimoxazole (960 mg, four times a day for 10 days). A complete response was obtained in the patient with concurrent cryptosporidiosis. The diarrhoea recurred after cessation of treatment, necessitating maintenance therapy. CONCLUSION: Although Isospora belli is an uncommon cause of gastroenteritis in Australia, it is increasingly recognised as a cause of diarrhoea in travellers, immigrants from endemic areas and immunocompromised people. It has also been reported in the Aboriginal community; this article reports the first case of AIDS-related isosporiasis in an Aborigine. It is likely that both patients acquired Isospora belli before they become HIV seropositive.

Acquired Immunodeficiency Syndrome

[Comparative review of the developmental biology of the genera Sarcocystis, Frenkelia, Isospora, Cystoisospora, Hammondia, Toxoplasma and Besnoitia (author's transl)].

A review is given of the advances in our knowledge of the developmental biology of the so-called cyst-forming coccidia in the years from 1974 to 1978. Until 1970 only 6 Isospora species were known to occur in cats, dogs and men. After the discovery of the coccidian nature of the genera Toxoplasma, Sarcocystis, Besnoitia and Frenkelia, and after the discovery of the new genus Hammondia the number of known species rose to over 30. In addition it could be shown that also birds of prey, owls and reptiles serve as final hosts for several Sarcocystis and Frenkelia species. The coccidia with isosporoid oocysts can be classified into two major groups: Species with gamogony and sporogony in the final host (Sarcocystis, Frenkelia) and species with schizogony and gamogony in the final host and sporogony on the ground (Isospora, Cystoisospora, Hammondia, Toxoplasma, Besnoitia). The subdivision of the first group into the genera Sarcocystis and Frenkelia based on the localization of their cysts in the musculature and in the brain, respectively, cannot be upheld in the future. Their classification into organisms with small cystozoites of about 7 microm with birds or reptiles as final hosts (Sarcocystis and Frenkelia species of rodents) and those with large cystozoites of about 15 microm and mammals as final hosts (Sarcocystis spp. of domestic animals and rodents) would be more significative. The second group can be subdivided into monoxenous species (Isospora), species with an optional intermediate host in which no or only slight multiplication occurs (Cystoisospora) and in genera with a multiplication in two phases in the intermediate host (Hammondia, Toxoplasma, Besnoitia). The nomenclature of single species is very controversial. As an example the controversial apprehension of the taxonomy of the Sarcocystis species of cattle is discussed. An application has been submitted to the International Commission for the Zoological Nomenclature to delcare a number of names as nomina dubia and to introduce unambiguous names for those organisms for which type specimens are available.

Animals

Life cycle of Isospora rivolta (Grassi, 1879) in cats and mice.

The endogenous development of Isospora rivolta (Grassi) was studied in cats fed oocysts, and was compared with the endogenous cycle after feeding them mice infected with I. rivolta. For the mouse-induced cycle, 14 newborn cats were killed 12 to 240 h after having been fed mesenteric lymph nodes and spleens ofmice. Asexual and sexual development occurred throughout the small intestine, in epithelial cells of the villi and glands of Lieberkühn. The number of asexual generations was not determined with certainty, but there were at least 3 structurally different meronts. Type I meronts appeared at 12-48 h postinoculation (HPT). They were 8.5(6-13) x 5.1(3-6) micrometer, contained 2-8 merozoites, and divide by binary division or endodyogeny. Type II meronts were multinucleate merozoite-shaped meronts within a single parasitophorous vacuole. They were found at 48-172 HPI and measured 12.6(9-18) x 9.8(9-13) micrometer. Individual multinucleate merozoite-shaped meronts were 7-13 x 3-5 micrometer in sections and contained 2-30 slender (5.5 x 1.0 micrometer) merozoites. Type III meronts occurred at 72-192 HPI and gamonts at 72-96 HPI. Mature microgamonts measured 11.3(9-15) x 8.0(6-9) micrometer in sections and up to 21.5 x 14 micrometer in smears, and contained up to 70 microgametes. Macrogamonts measured 13.3(11-18) x 9.0(5-13) micrometer in sections and 18 x 16 micrometer in smears, and contained up to 70 microgametes. Macrogamonts measured 13.3(11-18) x 9.0(5-13) micrometer. Sporulation was completed within 24 h at 22-26 C. For the study of the oocyst-induced cycle in cats, 18 newborn cats were killed between 6 and 192 HPI. The endogenous development was essentially similar to the mouse-induced cycle, but merogony and gametogony occurred 12-48 h later than in the latter cycle. Isospora rivolta was pathogenic for newborn but not for weaned cats. Newborn cats fed 10(6) sporocysts or infected mice usually developed diarrhea 3-4 days after inoculation. Microscopically, desquamation of the tips of the villi and cryptitis were seen in the ilium and cecum in association with meronts and gamonts. For the study of the development of I. rivolta in mice, mice were killed from day 1 to 23 months after having been fed 10(5)-10(6) sporocysts, and their tissues were examined for the parasites microscopically, and by feeding to cats. The following conclusions were drawn. (A) Isospora rivolta most freqeuntly invaded the mesenteric lymph nodes ofmice and remained there for 23 months at least. Ii also invaded the spleen, liver, and skeletal muscles of mice. This species could not be passed from mouse to mouse. Sporozoites increased in size from approximately 6.8 x 4.9 micrometer on day 1 to approximately 13.4 x 6.9 micrometer on day 31 postinoculation. Division was not seen. Prepatent period was 4-7 days and patent periods ranged from 2 to several weeks.

Animals

[Isospora belli infection in a patient with adult T-cell leukemia].

An adult T cell leukemia (ATL) accompanied with Isospora belli infection was described. A 65-year-old male was admitted to our hospital because of a two month history of watery diarrhea. On admission, physical examination showed slight pallor but no detectable superficial lymphadenopathies. Hepatosplenomegaly was not observed. Laboratory examination revealed a leukocyte count 5,500/microliters with 10% abnormal lymphoid cells. A majority of the abnormal lymphoid cells expressed both CD 4 and CD 8 antigens. The patient was diagnosed as chronic ATL, since anti-HTLV-1 antibody in his serum and monoclonal integration of HTLV-1 proviral DNA in his peripheral mononuclear cells were detected. Isospora belli was found in his feces thereafter, and trimethoprim/sulfamethoxazole was effective for diarrhea. In Japan, there have been only 9 reported cases of lymphoproliferative disorders (including five ATL patients) accompanied with Isospora belli infection. From the descriptions in those reports, these 9 cases might all be ATL patients.

Aged

Life cycle of Isospora burrowsi n sp (Protozoa: Eimeriidae) from the dog Canis familiaris.

Exogenous and endogenous stages of Isospora burrowsi n sp from the dog are described. Oocysts were spherical or ellipisoidal and 17 to 22 by 16 to 19 micrometer (mean 20.3 by 17.3 micrometer). Micropyle, oocyst residuum, and polar granule were not present. Sporocysts were 12 to 16 by 8 to 11 micrometer (mean 14.4 by 9.7 micrometer). Stieda body was absent, but a sporocyst residuum was present. Endogenous stages were in the caudal three-fifths of the small intestine and in the cecum. Mature 1st-generation schizonts (11 to 18 by 9 to 18 micrometer) were present on the 4th day of infection. One day later, mature 2nd-generation schizonts (18 to 35 by 17 to 22 micrometer) were present. Immature gamonts were found 5 days after infection, and unsporulated oocysts were also present later that day. The prepatent period was 6 days, and the mean patent period was 11 days. The oocysts of I burrowsi differ markedly in size from all other Isospora species from dogs, except Isospora ohioensis; however, the endogenous stages of the 2 species differ.

Animals

Isospora neorivolta SP. N. from the domestic dog.

The endogenous development of canine Isospora rivolta (Mahrt, J Protozool 14: 754--759, 1967) was compared with the development of I. ohioensis (Dubey, Parasitology, 1978, In press) in intestines of dogs. A new name, Isospora neorivolta, is proposed for the canine I. rivolta because of developmental differences from I. ohioensis. The major difference between these 2 coccidia is their site of development. Isospora neorivolta develops predominantly in the lamina propria of the posterior half of the small intestine, whereas I. ohioensis develops only in the epithelium and infection occurs throughout the small intestine. Additional information on the development of I. neorivolta in dogs is given.

Animals

Isospora gallicolumbae sp. n. from Beccari's ground dove (Gallicolumba beccarii Salvadori) in Papua New Guinea.

Isospora gallicolumbae sp. n. is described from Beccari's ground dove, Gallicolumba beccarii Salvadori, in Papua New Guinea. The ellipsoidal oocysts average 16.0 times 20.0 micron. The oocyst wall is light green, single-layered (approximately 0.8 micron), and becomes distorted within 10 min in sugar solution. Micropyle and oocyst residuum are absent; one polar granule is present. Sporocysts are ovoid, 8.0 times 12.0 micron, with prominent conical Stieda body. Sporocyst residuum is a spherical mass (approximately 5.0 micron) of lighter and darker granules. This is the first species of Isospora reported from a host in the avian order Columbiformes.

Animals

Isospora corvi Ray, Shivnani, Oommen and Bhaskaran, 1952 from the common house crow (Corvus splendens Vieillot) of Selangor, peninsular Malaysia.

Faecal samples of 56 common house crows (Corvus splendens Vieillot) were collected from the Petaling Jaya and Kelang districts of Selangor, peninsular Malaysia, and examined for coccidia. Intestinal tracts of 8 of the above crows wee histologically examined under light microscopy to determine the site of coccidial infection and the endogenous stages present. Fifty three (94.6%) crows had coccidial oocysts morphologically conforming to only one species of Isospora in their faeces at the time they were examined. The sporulated oocysts were found to be Isospora corviae (Ray et al. 1952) which has been emended to I. corvi. These oocysts are redescribed in greater detail. Corvus splendens is a new host record for I. corvi. Coccidial infection was observed in all the intestinal tracts and generally confined to the anterior two thirds of the intestine. The parasites occurred within intestinal epithelial cells, located usually above the host cell nucleus. Developmental stages of both the asexual and sexual phases were found in the epithelium, and are deemed to be the endogenous stages of I. corvi on the basis of the oocysts recovered from the same crows used for histological study. These stages are described here for the first time. The prevalence of I. corvi, its relationship with the host C. splendens, and its probable transmission from C. macrorhynchus are discussed.

Animals

[Utilization of the Kato-Katz-method (thick-smear technique) the diagnosis of Isospora (author's transl)].

Performing 10475 stool examination using the KATO-KATZ, HOFFMAN-PONS-JANER and direct examination (after agitation) methods, the author showed the best results with KATO-KATZ method for the diagnosis of Isospora hominis and Isospora belli. Among the 10475 stools examined 392 (3.74%) were positive for I. hominis and 55 (0.52%) for I. belli with the KATO-KATZ method; with the use of the others methods only 28 (0.26%) were positive for I. hominis and 6 (0.06% for I. belli.

Animals

Extraintestinal stages fo Isospora ohioensis from dogs in mice.

The development of Isospora ohioensis was studied in mice by feeding tissues of mice inoculated with oocysts to coccidia-free dogs and by the examination of mesenteric lymph nodes using light and electron microscopes. Extraintestinal organs of mice became infectious to dogs within 1 day after ingesting oocysts and remained infectious for at least 211 days after inoculation (DAI). Isospora ohioensis sporozoites were found in lymphoreticular cells of mesenteric lymph nodes of mice from 1-374 DAI. Intracellular sporozoites were located in parasitophorous vacuoles. Sporozoites grew from 5--6 to 11--16 micron in length on the 39th DAI but never lost the 2 crytalloid bodies typical for coccidian sporozoites. PAS-positive granules accumulated gradually in intracellular sporozoites with duration of infection in mice. The appearance of parasitophorous vacuoles varied with duration of infection. Beginning with 7 DAI, the vacuole contained a marginal zone of electron-dense material (up to 0.8 micron wide), giving the appearance of a cyst wall or sheath under the light microscope; a true cyst wall was was not found.

Animals

Malignant lymphoma initiated with malabsorption syndrome due to Isospora belli infection and lymphocytosis.

A 47-year-old man had diarrhea in 1965. Four years later, malabsorption syndrome was diagnosed and the patient was found to have mild lymphocytosis. Abdominal lymphoma was suspected, but exploratory laparotomy was normal except for partial villous atrophy of small intestine and slightly enlarged mesenteric lymphnodes which were normal microscopically. In vitro lymphocyte blastformation with phytohemagglutinin was depressed markedly throughout the course and the result predicted the developement of malignancy of the lymphocytic system. Infection of Isospora belli was found thereafter, and sulfamethoxazole was quite effective for diarrhea. In August, 1974, he noticed cervical lymphadenopathy for the first time and it was diagnosed as undifferentiated type of malignant lymphoma. He died in December, 1974. In this case diarrhea was most probably caused by the intestinal infection of Isospora belli without obvious lymphoma. The symptom was swept away by peroral sulfamethoxazole. In this patient coccidiosis was presumably induced and prolonged by suppression of cellular immunity which might have already begun to progress at the onset of diarrhea.

Biopsy

Electron microscopy of stages of Isospora felis of the cat in the mesenteric lymph node of the mouse.

Stages of Isospora felis of the cat in the mesenteric lymph node of the mouse 25 days after oral inoculation with oocysts, have been described at the ultrastructural level. The organisms occurred singly within parasitophorous vacuoles in host cell cytoplasm and were sporozoite-like, having a large crystalloid body up to 5.5 mum in length posterior to the nucleus. The size and appearance of the parasitophorous vacuole varied. Some vacuoles contained numerous, small, electron dense granules about 30 nm in diameter. Because of the aggregation of granules and their arrangement within the parasitophorous vacuole, the impression was sometimes gained by light microscopy that parasites were surrounded by a sheath or cyst wall. However, a cyst wall was not present. In host cells, spherical, membrane-bound bodies with a homogeneous, electron dense core and a maximum diameter of 0.25 mum were filed along the limiting membrane of the parasitophorous vacuole. These extra-intestinal parasites were considered to be waiting stages, with a biological function similar to that of the tissue cyst stage of other general of isosporan coccidia.

Animals