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[Light and electron microscopic studies on cysts of Sarcocystis fusiformis in the muscles of calves infected experimentally with oocytes and sporocysts of the large form of Isospora bigemina from dogs. 2. The fine structure of cyst stages].

In several experiments calves were infected with sporocytes of the large form of Isospora bigemina from dogs thus producing "thin-walled" cysts of Sarcocystis fusiformis in muscles. Following the growth of the cyst the development of the cyst stages (metrocytes, merozoites) was studied by electron microscopy. Cyst formation began about 34 days p.i. from a parasitophorus vacuole containing exclusively ovoid metrocytes. One the 62nd day p.i. mainly metrocytes and a few banana-shaped stages were present in the cysts. These banana-shaped stages were called merozoites, because we consider the process of cyst formation as an extraintestinal schizogony. From the 76th day p.i. mainly merozoites, i.e. infectious stages, were observed. The metrocytes were surrounded directly by the amorphous ground substance of the cyst's interior, whereas the merozoites were closely packed within chamber-like hollows of the ground substance. The metrocytes are globular cells, about 7 X 5 mum. These cells possess as the metrocytes of other species a typical three-layered pellicle with deep micropores, a conoid, polar ring with 22 anchored subpellicular microtubules, very few rhoptries and micronemes, a golgi complex anterior to the large nucleus. The nucleus has a spherical nucleolus consisting of granular and fibrillar zones. Chromosomal structures were seen in two different stages: large dense plaques (condensed stage), and as small dense granules of 300-400 A diameter, arranged spherically within the karyoplasm (extended stage). There is no significant difference in fine structure between the metrocytes of I. hominis-induced cysts and those from cysts after infections with the large form of I. bigemina from dog.

Animals↗

[Light and electron microscope studies on cysts of Sarcocystis fusiformis in the muscles of calves infected experimentally with oocysts and sporocysts of the large form of Isospora bigemina from cats (author's transl)].

In several experiments young calves were infected with oocyss and sporocysts of the large form of isospora bigemina from cats, which has been fed by raw beef containing Sarcocystis fusiformis cysts. On he 98th and 160th day p.i. the calves were killed and the development of S. fusiformis cysts in muscle cells was sutided by lign and electron microscopy. The cyst was always situated within a muscle fiber which was never surrounded by fibrillar layers of host origin (= no secondary cyst wall). The cyst was limited by a singel unit membrane, which was thickened at numerous places of the interior by osmiophilic material. This complex is called primary wall (Primärhülle), reaching a thickness of up to 250 A. This primary wall was regulary folded, forming palisade-like protrusions...

Animals↗

Isospora sp from ducks. Infectivity for the goose, four anatids and the domestic fowl.

A coccidium was isolated from mule duck farms, the sporulated oocysts of which most closely resemble that of Isospora anseris, a goose coccidium. By experimental infection, these oocysts were proved to be able to develop in the goose and in several duck species, but unable to develop in the chicken. No pathogenicity was noted in any species.

Animals↗

Ultrastructure of the sporocyst wall during excystation of Isospora endocallimici.

Sporocysts of Isospora endocallimici, a parasite of marmosets, were exposed to minimal essentials medium (MEM) or a trypsin-bile salt solution (TBS) and then fixed and prepared for transmission electron microscopy. Excystation occurred in TBS but not MEM. The sporocyst wall has 2 layers, a thin outer layer (15 to 110 nm thick) and a thick inner layer (65 to 180 nm thick), which is composed of 4 separate curved plates. The outer layer consists of 1 to 3 membranes interspersed with lipid droplets. In the inner layer, a thin layer of material connects the peripheral margins of 2 apposing plates. Immediately beneath this layer, a thin strip of material is interposed between the 2 apposing plates. Ultrastructural changes preparatory to excystation occur primarily in the inner layer of the sporocyst wall. The TBS acts upon the site of apposition between 2 plates causing the interposed strip to swell and separate from the margin of each plate which leads to collapse of the sporocyst. As the sporocyst collapses, the margins of each curved plate curl inward toward the center of the sporocyst.

Animals↗

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

A 53-year-old man was hospitalized for the development of watery diarrhea associated with decreased appetite and progressive weight loss and was found to have leukocytosis. The white blood cell count was 14,600/microliters with 66 percent abnormal lymphocytes. Serum anti-HTLV-I was positive and monoclonal insertion of HTLV-I provirus into the atypical cell genome was confirmed with the southern blotting hybridization technique. A diagnosis of ATL was made. Examination of fresh stool specimens revealed Isospora belli (I. belli) oocysts. Initial treatment for I. belli consisted of oral trimethoprim 160mg and sulfamethoxazole 400mg given twice daily for nine days. Diarrhea ceased within 2 days of the start of treatment, but I. belli oocysts were again detected after 20 days. Trimethoprim 160mg and sulfamethoxazole 400mg were reinstituted four times daily for 10 days, and then twice daily for 21 days. The clinical response was again dramatic, with rapid clearance of oocysts from the stool. There has been no recurrence of diarrhea. The patient's leukemia was refractory to chemotherapy; the white blood cell count continued to rise, pneumonia developed, and the patient died. I. belli is a previously unrecognized opportunistic pathogen that must be considered in the clinical setting of chronic diarrhea in patients with ATL.

Animals↗

Prevalence of Cryptosporidium sp. and Isospora belli among AIDS patients attending Santos Reference Center for AIDS, São Paulo, Brazil.

From March 1991 to February 1992, 276 stool samples from 131 patients with the acquired immunodeficiency syndrome (AIDS) (average, 2.1 samples per patient) and stool samples from 81 presumed immunocompetent individuals were studied for the prevalence of Cryptosporidium sp. and Isospora belli. The techniques employed were a modified formol-ether concentration, carbol (phenol) auramine staining, and a modified Kinyoun acid-fast method. The prevalence of both coccidia among AIDS patients was 25.9% (34 of 131). Cryptosporidium sp. oocysts were demonstrated in 25 (19.1%) AIDS patients and I. belli in 13 (9.9%). Coinfection for coccidia of both species was found in 4 (3%) AIDS subjects. Among the presumed immunocompetent individuals, coccidia were not demonstrated in their 81 stool specimens. This study, like others, confirms the worldwide importance of these coccidia among AIDS patients and the necessity of suitable techniques for demonstration of oocysts in stool samples.

AIDS-Related Opportunistic Infections↗

Isospora ashmoonensis sp. nov. from blackcap warbler, Sylvia atricapilla in Egypt.

Microscopic examination of faecal samples collected from naturally infected blackcap warbler, Sylvia atricapilla revealed the presence of a new Isospora species; I. ashmoonensis. Oocysts were subspherical to oval, measuring 24.0 (21.0-25.0) microns in length and 20.0 (19.0-20.5) microns in width. A micropyle and an oocyst residuum were absent whereas one polar granule was seen. Ellipsoidal sporocysts contained a stieda body and measured 17.0 (16.0-18.5) x 9.2 (9.0-10.0) microns. Sporulation took place within 50 hrs. at room temperature. Developmental stages of the parasite in epithelial cells of the posterior third of the ileum were also described.

Animals↗

[Prevalence of Cryptosporidium sp and Isospora belli in patients with acquired immunodeficiency syndrome (AIDS) in Dakar (Senegal].

72 patients with AIDS, having diarrhoea and admitted in the Unit of Infectious diseases, Fann Hospital from 1989 to 1991 were investigated for Cryptosporidium sp. and Isospora belli isolation by the modified Ziehl-Neelsen technique, after stolls' concentration by Ritchie technique. The prevalence were 13.9% for Cryptosporidium sp, and 15.3% for I. belli. In two patients (2.8%), these 2 coccidiae were associated. Cryptosporidium sp. and I. belli were the single parasites found respectively in 6 (8.3% and 7 (9.7%) of the patients. In the other faecal samples with a positive result for Cryptosporidium sp. or I. belli, they were associated with A. lumbricoides, G. intestialis, E. histolytica or S. stecoralis. According to these results, Cryptosporidium sp. and I. belli were more frequently observed in AIDS patients from Dakar than those from Zaĭre, Congo and Côte d'Ivoire.

Acquired Immunodeficiency Syndrome↗

Cryptosporidium, isospora and strongyloides in AIDS.

BACKGROUND: Diarrhoea is a common presentation in patients with AIDS. It occurs due to a number of parasites which are seldom seen in immunocompetent hosts. METHODS: Between January 1993 and July 1994, faecal specimens from 77 patients with AIDS presenting with diarrhoea of more than a month's duration were examined by direct wet mount microscopy of saline and iodine preparations and by the modified Ziehl-Neelsen stain. RESULTS: Cryptosporidium, Isospora and Strongyloides stercoralis alone or in combination were present in 29 of the 77 patients. CONCLUSION: The presence of these parasites highlights their pathogenic potential for immunocompromised patients with AIDS. Diarrhoea due to Isosporais responsive to treatment, hence their identification has therapeutic implications as well.

AIDS-Related Opportunistic Infections↗

Light and electron microscopic studies of Isospora chalchidis (Amoudi, 1989) infecting Chalcides ocellatus in Egypt.

Oocysts of Isospora chalchidis (Amoudi, 1989) were described from the faeces of the skink Chalcides ocellatus in Egypt. Non-sporulated oocysts were spheroidal, measuring 20 (18.5-21) microns in diameter and contained granulated zygotes. Sporulated oocysts had the same dimensions of the nonsporulated ones and each contained two sporocysts. Sporocysts were ovoid with stieda and sporocyst residual bodies. Sporulation time was 50 hours at room temperature. Merogony and gamogony occurred in the intestinal mucosa. Electron microscopic investigations showed that meronts, merozoites, gamonts and gametes developed in a narrow parasitophorous vacuole within the host-cell nucleus. Nuclei of meronts were surrounded by rough endoplasmic reticulum. Merozoites showed the main characteristics of motile stages of Apicomplexa. Macrogametes contained a large nucleus, two types of wall-forming bodies and a large amount of lipid inclusions. Nuclei of microgamonts were peripherally arranged and lacked nucleoli. Microgametes were flagellated.

Animals↗

[The epizootiology and pathogenic significance of infections with Isospora suis in ten piglet production operations in Nordrhein-Westfalen].

From July 1993 to June 1994 2036 fecal samples from 196 litters from 10 pig farms were collected. Three or four piglets from each litter and the respective sows were examined 3 times at 2, 3 and 4 weeks after farrowing. I. suis was detected in all 10 farms. The rate of infection of the litters was 62.2%. 71.6% of all litters with diarrheic problems were I. suis positive. The percentage of I. suis in the feces of piglets was 17.4% at the age of 2 weeks, 41.3% at the age of 3 weeks and 39.1% at the age of 4 weeks. It was also shown that I. suis occurs on any farm with any type of management system and at any time of the year. Fecal samples of sows were always Isospora negative. It is inferred that I. suis is a more important causative agent of porcine neonatal diarrhoea than considered so far. Therefore, further studies on epizootiological and therapeutical aspects and sanitation measures are necessary.

Animals↗

Gut Coccidia--Isospora, Cryptosporidium, Cyclospora and Sarcocystis.

The gut Coccidia are members of a large, varied, and exclusively intracellular group of protozoan parasites, four species of which (Isospora, Cryptosporidium, Cyclospora, and Sarcocystis) are human pathogens. The first three, but particularly Cryptosporidium parvum, have moved from medical curiosities to major problems with the coming of the acquired immunodeficiency syndrome (AIDS) epidemic, but are now known to also cause disease in the immunocompetent patient. They are easy to acquire and difficult to remove from the environment and, in the case of cryptosporidiosis, impossible to treat properly. Further research into many aspects of the biology of these organisms is urgently needed.

Animals↗

Isospora tiaris n.sp. (Apicomplexa: Eimeriidae) from the sooty grassquit (Tiaris fuliginosa), a passeriform bird of South America.

Isospora tiaris n. sp. is described from the feces of an adult bird, a sooty grassquit (Tiaris fuliginosa) from Venezuela. The oocysts are subspherical, 27.1 x 23.8 (24.7-30.0 x 21.2-26.5) microns with a bilayered, smooth, colorless wall. A micropyle and oocyst residuum are absent; an ellipsoidal polar granule is usually present. Sporocysts are ovoidal, 14.7 x 10.8 (12.4-16.8 x 8.8-12.4) microns with prominent Stieda and substieda bodies and a residuum composed of small uniform granules.

Animals↗

A new Isospora sp. from Carduelis tristis (Aves: Fringillidae) from Ontario, Canada.

Isospora gryphoni n. sp. is described from oocysts found in the feces of the American goldfinch, Carduelis tristis, from Guelph, Ontario, Canada. The oocysts are spherical to subspherical, with a double-layered, smooth, colorless oocyst wall, 29.2 x 30.7 microm (25-33 x 28-34; n = 30) with 2-4 rice-grain-shaped polar bodies; no micropyle or residuum. Sporocysts are ovoid, 22.2 x 13.4 microm (15-25 x 12-14.5; n = 30) with a small Stieda body, indistinct substiedal body, and prominent sporocyst residuum. Sporozoites are vermiform, each with a large refractile body at the posterior end. Forty-nine of 52 (94%) wild-caught adult C. tristis and 1 juvenile bird (minimum 12 days old) were excreting oocysts of I. gryphoni. None of 36 nestlings (maximum 10 days old) was excreting oocysts. Fecal encrustations from 7 of 10 used nests were found to contain oocysts of I. gryphoni.

Animals↗

Complete development of the porcine coccidium Isospora suis Biester, 1934 in cell cultures.

Development from inoculated sporozoites to unsporulated oocysts of Isospora suis Biester, 1934 is described in a swine testicular (ST) cell line. Sporozoites penetrated ST cells within 1 hr postinoculation (PI). Development was initially by endodyogeny to produce binucleate type I meronts and type I merozoites. Division by endodyogeny continued during the 13-day observation period and type I merozoites were the developmental stages most abundant at observation periods >3 days PI. Mutinucleate type II meronts and type II merozoites were first observed 7 days PI. Gamonts and oocysts were present 12 days PI. Oocysts did not sporulate in vitro. The ultrastructural features of stages were similar to those that occur in the pig host.

Animals↗

DNA circles with cruciforms from Isospora (Toxoplasma) gondii.

We have isolated a closed circular duplex DNA fraction from the unicellular parasite Isospora (Toxoplasma) gondii and examined the purified DNA by electron microscopy. A major part of this circular DNA consists of 12-micron circles containing a cruciform with 0.5-micron tails. We also found 23-micron circles with the properties expected of head-to-tail dimers of the 12-micron circles. Some of these dimers have two cruciforms with 0.4-micron tails, some have one cruciform with 0.8-micron tails. When ethidium bromide was diffused into the DNA solution, circles with tails were replaced by twisted circles without tails. Direct mixing of the DNA with high ethidium bromide concentrations (5 micrograms/ml) gave rise to highly twisted circles with tails. This proves that the tailed circles are covalently continuous and indicates that ethidium bromide blocks branch migration. The 0.5-micron tails are part of a 1.7-micron palindrome, which was visualized by spreading denatured DNA under snap-back conditions. We argue that the cruciform is not present in vivo and that the 12-micron circles may represent the mitochondrial DNA of Toxoplasma.

DNA, Bacterial↗

Experimental Isospora suis infections in miniature swine.

Clinical responses to experimental Isospora suis infections were compared in Sinclair miniature pigs and cross-bred conventional pigs. Pre-patent periods, fecal consistencies, oocyst excretion dynamics, trends in surviving pig weights and lesions were similar in infected miniature and conventional pigs. The results indicate that the susceptibility of miniature pigs to I. suis is similar to that of conventional pigs. These findings should encourage their use as models for the study of neonatal coccidiosis.

Animals↗

Isospora (Toxoplasma) gondii: induction of sexuality.

The role of the stomach and small intestine of cats and of the parasite's rate of multiplication on induction of sexuality in Isospora (Toxoplasma) gondii was studied by injecting cats either with free cystozoites or proliferative parasites directly into the intestinal lumen, after laparotomy. Oocyst production was similar in cats infected orally with cysts or cystozoites and in cats infected by inoculation into the duodenal lumen with free cystozoites, obtained by either mechanical rupture of by in vitro pepsin/HCl digestion of the cyst wall. When free cystozoites were injected into the lumen of the posterior part of the ileum, cats became seropositive but oocyst excretion during the first 4 weeks after infection was very low or absent; nevertheless a solid immunity to oral challenge was acquired in the former case. If no oocysts at all were produced after primary infection, the intestine remained susceptible to challenge, in spite of previous seroconversion. This lack of protective immunity in the presence of serum antibodies was observed in all cats primarily infected by direct injection of proliferative parasites into the duodenal lumen. It is concluded that previous gametogony rather than previous infection and seroconversion provokes local intestinal immunity against the development of sexual stages after oral challenge; the switch to gametogony of cystozoites is not triggered off by the low pH of the stomach but is probably related to their reduced rate of multiplication.

Animals↗