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[A clinical and biological study of parasitic and fungal diarrhea in immunosuppressed patients in an urban and suburban area of Yaoundé].

We studied 66 cases of intestinal mycosis and parasitosis in patients infected with the human immunodeficiency virus with chronic diarrhea. All subjects were from the Yaounde urban area and were followed between February and December 1996. They were recruited from 3 hospitals in the center of Yaounde and were aged between 2 and 52 years. There was weight loss in 80.3% and severe dehydration in 72.3% of cases. Feces consisted mostly of watery stools similar to those of cholera patients (50% of cases) and loose stools (43.9% of cases). Parasitic agents were detected in 31.8% and fungal agents in 80.5% of cases. The opportunistic Protozoans detected included Cryptosporidium parvum (15.8%), microsporidia (8.8%) and Isospora belli (3.5%). Six cases of helminthiasis were also identified. Candida albicans was the most common opportunistic mycosis agent (39.1%). Other fungal species detected included Geotrichum candidum, Candida pseudotropicalis and Trichosporon sp. but all of these were less common.

AIDS-Related Opportunistic Infections↗

[HIV infection and isosporiasis. Presentation of a case].

A HIV seropositive patient with chronic diarrheic syndrome, in which oocysts of Isospora belli were detected in feces by the modified technique of Ziehl-Neelsen was studied. Other coproparasitological techniques were applied and cultures were applied made with negative results. Any other infectious origin was excluded from the picture. The patient was treated with 2 tablets of Co-trimoxazole (trimethoprin 80 mg, sulphamethoxazole 400 mg) every 6 hours for ten days, with referral of the symptoms.

AIDS-Related Opportunistic Infections↗

Two new coccidian parasites from the grand anglehead lizard, Gonocephalus grandis from Peninsular Malaysia.

During 3 collecting expeditions between October 1996 and December 1996, fecal samples were obtained from 43 adult Gonocephalus grandis from Tanah Rata and the Cameron Highlands in Peninsular Malaysia. Two species of coccidia (Isospora gonocephali n. sp. [9/43, 23%] and Eimeria cameronensis n. sp. [3/43, 7%]) were discovered. Sporulated oocysts of I. gonocephali are subspherical to ovoidal, 22.3 x 18.7 (19-25 x 17-23) microm with a bilayered wall composed of a thin inner wall and a striated outer wall with a pitted surface; oocyst residuum absent; 1 polar granule present; sporocysts are almond-shaped, 13.5 x 9.2 (12-15 x 8.5-10) microm, Stieda body broad, domelike, substieda body fanlike, sporocyst residuum consisting of coarse, nonuniform granules in an amorphous cluster; sporozoites sausage-shaped with 1 large terminal, refractile body and lay randomly in the sporocyst. Sporulated oocysts of E. cameronensis are bilayered, smooth-walled, ellipsoidal, 26.5 x 12.4 (25-28 x 12-13) microm; with 1, small, polar granule composed of 2-3 splinter-like structures fused together; oocyst residuum absent; sporocysts ovoidal, almost rectangular-shaped 8.8 x 6.6 (8-9 x 5-7) microm, with no Stieda or substieda bodies, containing scattered residuum and 2 sausage-shaped sporozoites with 1 terminal, ovoidal refractile body. No individual lizard was host to both coccidian species.

Animals↗

Protozoal infections in the acquired immunodeficiency syndrome.

Several protozoa have emerged as the major opportunistic infections and cause of death in patients with acquired immunodeficiency syndrome (AIDS). Pneumocystis carinii pneumonia is the leading cause of death in AIDS patients. Electron microscopy (EM) usually shows numerous trophozoites and cysts of Pneumocystis filling up the entire alveolar space, while only cysts are seen under the light microscope. The focal thickening of cyst wall of Pneumocystis, as demonstrated by EM and manifested as a "parentheses" shaped structure with silver stain, serves as a diagnostic marker for Pneumocystis. Freeze-fracture EM has demonstrated the intimate contact between Pneumocystis trophozoites and the type I pneumocytes, which may contribute to the alveolar-capillary block, leading to severe respiratory distress. However, EM is seldom needed for the diagnosis of this infection. Toxoplasma encephalitis, which is an unusual clinical manifestation in cases of toxoplasmosis reported previously, has become a common complication and one of the major causes of death in patients with AIDS. Because subclinical infection by Toxoplasma is common, serologic tests usually offer no definite answers as to whether the infection is acute or chronic, active or past. The small size and its non-specificity in both morphology and tissue affinity make light microscopic diagnosis of toxoplasmosis difficult. Only immunologic staining, such as immunoperoxidase and immunofluorescence, can help to achieve a definite positive identification of the organism. When special antibodies or facility for such staining is not available, EM is the final resort for identifying Toxoplasma by showing the apical complex with the characteristic sausage-shaped rhoptries. Cryptosporidiosis, practically unknown before the AIDS outbreak, has become one of the most common intestinal protozoa in both immunocompromised and immunocompetent patients. The protracted and sometimes fatal course of cryptosporidiosis in immunocompromised patients can be explained by the presence of autoinfective oocysts (thin-walled oocysts), as detected by EM, and by recycling of first-generation schizonts observed experimentally. While diagnosis of cryptosporidiosis can be made by detection of oocysts in stools in most cases, EM is still the last resort for a definitive identification of Cryptosporidium species. While the incidence of isosporiasis is still low, it has been found more frequently in patients with AIDS than in the general population. The parasite, Isospora belli, being a coccidian as is the Cryptosporidium species, is similar to the latter in its life cycle and clinical manifestations.(ABSTRACT TRUNCATED AT 400 WORDS)

Acquired Immunodeficiency Syndrome↗

Parasitophorous vacuole: morphofunctional diversity in different coccidian genera (a short insight into the problem).

We present a short insight into the problem of parasitophorous vacuole (PV) formation as a most peculiar kind of cell vacuolization occurring in the course of intracellular development of coccidian pathogens of the genera Eimeria, Isospora, Toxoplasma, Sarcocystis, Cryptosporidium, Epieimeria, and Karyolysus. The review focuses on the morpho-functional diversity of PVs in these parasites. By the present time, the PVs containing different parasite genera and species have been examined to different extent. The membrane of the PV (PVM) obviously derives from the host cell plasmalemma. But soon after parasite penetration, the morphofunctional organization and biochemical composition of the PVM drastically changes: its proteins are selectively excluded and those of the parasite are incorporated. As the result, the PV becomes not fusigenic for lysosomes or any other vacuoles or vesicles, because host cell surface markers necessary for membrane fusion are eliminated from the PVM during parasite invasion. The pattern of the PVs is parasite specific and demonstrates a broad diversity within the same genera and species and even at different stages of the endogenous development. The PV is far from being an indifferent membrane vesicle containing the parasite. Instead, it represents a dynamic system that reflects the innermost events of host-parasite relationships, thus promoting the accomplishing of the parasite life cycle, which, in its turn, is a necessary prerequisite of the parasite eventual survival as a species.

Animals↗

Ribosomal RNA sequences for the specific detection of Toxoplasma gondii by hybridization assay.

We describe the development of five oligonucleotide probes which target the small sub-unit rRNA of Toxoplasma gondii. These probes bind to different regions within the rRNA and all were effective in producing a detectable signal with as little as 10 ng of total cellular RNA. One probe in particular, TGB, was able to detect as little as 1 ng of RNA and was the most specific. A sixth probe, previously developed for hybridization with Sarcocystis spp., was evaluated for its specificity and was shown to be useful as a multi-species coccidial probe. Four of the five T. gondii-specific probes did not cross-hybridize with RNA from nine related species of coccidia, Escherichia coli, cattle, people, cat or dog. One probe cross-hybridized strongly with RNA of Eimeria ahsata and Isospora suis, but not to RNA of any other coccidia species tested nor bovine, human, feline or canine RNA. The potential of this assay for use in veterinary and human medicine as well as food safety programmes is discussed.

Animals↗

Pneumocystis carinii: taxing taxonomy.

The detailed taxonomy of Pneumocystis carinii has not been decided. Most experts agree that it is either a fungus or a protozoan. In favor of a fungal designation are the recent reports that 16s and 18s rRNA sequences of P. carinii have greater homology to Saccharomyces cerevisiae and Neurospora crassa than to selected protozoa. Other studies show that sequences of the 18s and 26s rRNA, microtubular ultrastructure, membrane fusion potential and DNA content per cell resemble protozoa more than fungi. Therapeutic responsiveness to drugs effective against protozoa, namely pentamidine (Trypanosoma, Leishmania, sp), trimethoprim-sulfamethoxazole (Isospora sp), pyrimethamine-sulfadiazine (Toxoplasma, Plasmodium sp) and nonresponsiveness to antifungal drugs such as amphotericin B and ketoconazole indicate characteristics similar to protozoa. Taxonomic placement is based upon multifactional determinants, not a single attribute. Data are not yet available to make such a determination for P. carinii. Furthermore, detailed rules on qualifying criteria for both protozoan and fungal classifications are lacking, especially in the utilization of methods in molecular genetics for this purpose.

Animals↗

AIDS-related cholangiopancreatographic changes.

The cholangiographic and pancreatographic appearances of the acquired immunodeficiency syndrome (AIDS) associated cholangitis were evaluated in 26 patients. Twenty-four patients were diagnosed by retrograde cholangiography or endoscopic cholangiopancreatography (ERC or ERCP). One patient was diagnosed by T-tube cholangiography and another patient by transhepatic cholangiography. The radiographic findings ranged from intrahepatic ductal abnormalities with or without involvement of the extrahepatic biliary tree (eight patients) to irregularities and strictures involving the ampulla of Vater or the intrapancreatic portion of the common bile duct (CBD) with proximal dilatation (18 patients). Significant strictures involving the juxta-ampullary pancreatic duct were identified in six of 12 patients. Twenty-one of the 26 patients had associated infections which included: Cryptosporidium (CS), Mycobacterium avium intracellulare (MAI), cytomegalovirus (CMV), Microsporidium (MSP), and Isospora (ISP). Three patients were operated upon for acute acalculous cholecystitis. In each instance, organisms were identified in both the bile duct and the inflamed gallbladder.

AIDS-Related Opportunistic Infections↗

[Serological studies on Sarcocystis in man and rats (author's transl)].

During a study of sarcosporidiosis in man, rats too were fed on meat infected with Sarcocystis tenella and A. fusiformis. Further investigations were made on the antigen relationship between Sarcocystis and Toxoplasma using 50 mice infected with Toxoplasma. The results are as follows: 1. 18-39 days after eating raw minced meat (beef) containing Sarcocystis fusiformis, 5 (=41,7%) out of 12 people excreted sporulated sporocysts of the species Isospora hominis in the faeces. 2. Excretion of sporocysts lasted for 9-179 days without reinfection. None of the above mentioned people showed any clinical symptoms. 3. There was no manifestation of immunity as the excretion of sporocysts which had reduced in number intensified again after repeated administration of similar raw minced meat. 4. Oral administration of Sarcocystis as antigen leads to the production of antibodies which could be detected with the indirect immunofluorescent test (IIFT). These antibodies are common among adults but could be detected in children after 6 months of age. 5. Sarcocystis antibodies produced in rats after ingestion of fresh raw S. tenella-infected meat could be detected with the help of the IIFT in their sera for several months. 6. The indirect immunofluorescent test is reliable in detecting Sarcocystis infection. 7. No cross-reaction between Sarcocystis and Toxoplasma was detected in either the human sera or in the experimentally Sarcocystis-infected rats and Toxoplasma-infected mice. 8. Parasites lose their capability to function as antigen when subjected to heat or cold because rats fed on frozen or cooked S. tenella-infected mutton did not produce antibodies compared to those fed on fresh raw infected mutton. 9. Sera of rats fed on different types of sausages reacted negative with Sarcocystis antigen.

Adult↗

Light and electron microscopic observations of the life cycle of Sarcocystis orientalis sp. n. in the rat (Rattus norvegicus) and the Malaysian reticulated python (Python reticulatus).

A light and electron microscopic study of Sarcocystis orientalis sp. n. was made. The life cycle of this parasite is in two hosts. Gametogony is in the intestinal epithelial cells of a predator, Python reticulatus. Isospora-like oocysts developed. Sporocysts average 9.1 by 7.7 mum. Rats (Rattus norvegicus) were infected with sporocysts and asexual stages developed. Ten days after infection large zoites (average 7.85 by 2.48 mum) were observed free in peripheral blood and within white blood cells. Small schizonts producing merozoites 2-3 mum long were seen in lung tissue. Tissue cysts developed in skeletal muscle and produced numerous cystozoites (average 5.53 by 1.38 mum). Fine structure was similar to previously described Sarcocystis spp.

Animals↗

Sarcocystinae: nomina dubia and available names.

Examination of the original descriptions of the species of Sarcocystis in cattle, sheep, and swine, and of isosporid oocysts shed sporulated by dogs, cats, man, and other carnivores, has shown that it is not possible in most instances to identify unambiguously recently recognized taxa. The original descriptions are insufficient, and because no type specimens exist, could apply to two or more of the presently recognized taxa. We consider the following nomina dubia: Sarcocystis hirsuta S. miescheriana S. tenella S. cruzi S. bertrami Isospora bigemina (S. bigemina) I. hominis (S. hominis) I. buteonis (Frenkelia buteonis) Because the former type species, Sarcocystis miescheriana, is an indeterminate nomen dubium, we are proposing S. muris as the new type species. Historically, it was the first species described clearly and unambiguously even in the light of present knowledge, and the stages of its life cycle are probably completely known; it was the second species to be named. Old and recent descriptions are reviewed, and definitions are proposed for the following taxa: S. bovifelis S. bovicanis S. bovihominis S. ovifelis S. ovicanis S. muris (type species) S. suihominis S. suicanis S. equicanis Frenkelia microti F. glareoli for which neotypes will be prepared and deposited with designated institutions and curators. A new subfamily, Cystoisosporinae, is created.

Animals↗

Sexually transmitted diseases of the colon, rectum, and anus. The challenge of the nineties.

During the past two decades, an explosive growth in both the prevalence and types of sexually transmitted diseases has occurred. Up to 55 percent of homosexual men with anorectal complaints have gonorrhea; 80 percent of the patients with syphilis are homosexuals. Chlamydia is found in 15 percent of asymptomatic homosexual men, and up to one third of homosexuals have active anorectal herpes simplex virus. In addition, a host of parasites, bacterial, viral, and protozoan are all rampant in the homosexual population. Furthermore, the global epidemic of AIDS has produced a plethora of colorectal manifestations. Acute cytomegalovirus ileocolitis is the most common indication for emergency abdominal surgery in the homosexual AIDS population. Along with cryptosporidia and isospora, the patient may present to the colorectal surgeon with bloody diarrhea and weight loss before the diagnosis of human immunodeficiency virus (HIV) disease. Other patients may present with colorectal Kaposi's sarcoma or anorectal lymphoma, and consequently will be found to have seropositivity for HIV. However, in addition to these protean manifestations, one third of patients with AIDS consult the colorectal surgeon with either condylomata acuminata, anorectal sepsis, or proctitis before the diagnosis of HIV disease. Although aggressive anorectal surgery is associated with reasonable surgical results in some asymptomatic HIV positive patients, the same procedures in AIDS (symptomatic HIV positive) patients will often be met with disastrous results. It is incumbent upon the surgeon, therefore, to recognize the manifestations of HIV disease and diagnose these conditions accordingly.

Acquired Immunodeficiency Syndrome↗

Prevalence of intestinal pathogens in HIV patients with diarrhea: implications for treatment.

Patients infected with the human immunodeficiency virus (HIV) commonly experience diarrhea at some time during their illness. A variety of enteric pathogens are identified in 50-80% of these patients, depending on the intensity of the diagnostic work-up that is done. In addition to the common enteric pathogens, several unusual enteric pathogens are recognized to cause diarrhea especially in HIV patients. These include protozoan parasites such as Cryptosporidia, Isospora belli, Cyclospora cayatenensis and Microsporidium species bacteria such as enteropathogenic Escherichia coli and Mycobacterium avium-intracellulare, fungi including Candida albicans and Histoplasma capsulatum, and viruses such as astroviruses and caliciviruses. Diagnosis of these infections sometimes involves special procedures not readily available every where, and empiric therapy based on knowledge of the likely pathogens has been advocated for developing countries. This article reviews the currently available data on geographic variation of enteric pathogens in HIV patients with diarrhea and outlines a rational strategy for empiric therapy of these patients.

Animals↗

Phylogenetic relationships of Sarcocystis neurona of horses and opossums to other cyst-forming coccidia deduced from SSU rRNA gene sequences.

Phylogenetic analyses based on sequences of the nuclear-encoded small subunit rRNA (ssurRNA) gene were performed to examine the origin, phylogeny, and biogeographic relationships of Sarcocystis neurona isolates from opossums and horses from the State of Michigan, USA, in relation to other cyst-forming coccidia. A total of 31 taxa representing all recognized subfamilies and genera of Sarcocystidae were included in the analyses with clonal isolates of two opossum and two horse S. neurona. Phylogenies obtained by the four tree-building methods were consistent with the classical taxonomy based on morphological criteria. The "isosporid" coccidia Neospora, Toxoplasma, Besnoitia, Isospora lacking stieda bodies, and Hyaloklossia formed a sister group to the Sarcocystis spp. Sarcocystis species were divided into three main lineages; S. neurona isolates were located in the second lineage and clustered with S. mucosa, S. dispersa, S. lacertae, S. rodentifelis, S. muris, and Frenkelia spp. Alignment of S. neurona SSU rRNA gene sequences of Michigan opossum isolates (MIOP5, MIOP20) and a S. neurona Michigan horse isolate (MIH8) showed 100% identity. These Michigan isolates differed in 2/1085 bp (0.2%) from a Kentucky S. neurona horse isolate (SN5). Additionally, S. neurona isolates from horses and opossums were identical based on the ultrastructural features and PCR-RFLP analyses thus forming a phylogenetically indistinct group in these regions. These findings revealed the concordance between the morphological and molecular data and confirmed that S. neurona from opossums and horses originated from the same phylogenetic origin.

Animals↗

Efficacy of various anticoccidials against experimental porcine neonatal isosporosis.

The efficacies of 20 mg/kg body weight (BW) of toltrazuril (Toltra) 2 days post-infection (dpi), 2 mg/kg BW of diclazuril 2 and 3 dpi and 200 mg/kg BW of sulphadimidine 2, 3 and 4 dpi were compared in a model for piglet isosporosis. Weight gain (first 4 weeks of life) and diarrhoea and oocyst excretion from 4 to 11 dpi were evaluated (10-12 piglets/group). Additionally, animals were killed and examined for pathohistological changes of the small intestines 5, 7, 11 and 14 dpi (n = 3 per group and time point) and lengths of the intestinal villi. Diarrhoea (semi-liquid or liquid faeces) was seen from 5 dpi in all groups except Toltra, and the differences in prevalence and intensity of diarrhoea were statistically significant (p < 0.05) between Toltra and the other groups, which were similar (trial 1). Oocyst excretion was greatly reduced in the Toltra group, which was also statistically significant for the mean and median excretion rates and the percentage of excreting piglets between Toltra and the other groups (p < 0.05). Weight gain was highest in Toltra (p < 0.05). Histopathology revealed mostly villous necrosis and atrophy in the small intestines except the duodenum, which peaked at 7 dpi, in all groups except Toltra. Between 5 and 11 dpi, the Toltra group had significantly longer villi than the other groups. Reduced weight gain and diarrhoea caused by Isospora suis was controlled by a single application of Toltra in the pre-patent period, while neither diclazuril nor sulphadimidine improved the clinical picture of isosporosis.

Animals↗

Neospora caninum: is it really different from Hammondia heydorni or is it a strain of Toxoplasma gondii? An opinion.

The published data concerning Toxoplasma gondii, Hammondia hammondi, H. heydorni and Neospora caninum on one side and between T. gondii on the other were neglected by most authors. As conclusion we are convinced that there are only two valid species: Isospora (Toxoplasma) gondii and Hammondia heydorni. The first includes as a strain H. hammondi and the latter N. caninum. In any case there is absolutely no reason (with respect to general Zoological nomenclature) to create new genera!

Animals↗

Hammondia heydorni-like oocysts shed by a naturally infected dog and Neospora caninum NC-1 cannot be distinguished.

This study describes transmission experiments using Hammondia heydorni-like oocysts isolated in 1996 from a naturally infected dog. The isolate was designated as H. heydorni-Berlin-1996. Examination of sera from infected intermediate hosts showed immunoblot reactions that resembled patterns observed after Neospora caninum NC-1 infection. Furthermore, N. caninum DNA could be demonstrated in tissue samples (e.g. heart, brain) of experimentally infected intermediate hosts and in oocyst preparations from H. heydorni-Berlin-1996. The isolated oocysts did not induce any detectable disease in any of the inoculated adult intermediate hosts (goats, sheep, gerbils, guinea pigs, multimammate rats, BALB/c mice, SCID mice), even upon immunosuppression. Furthermore, neither histological lesions nor parasite stages could be identified in the tissues of all fetuses recovered from two multimammate rats that had been infected prior to pregnancy. An experiment with one dog fed a second time on infected intermediate host tissue indicated that immunity may prevent repeated oocyst shedding in N. caninum-infected dogs. In addition, the study clearly demonstrates that N. caninum can be readily transmitted by dogs that have ingested exclusively skeletal muscles of infected intermediate hosts. Therefore, the study has consequences for the recommendations for farmers to prevent postnatal transmission of N. caninum to cattle. It indicates that feeding of any tissues of potential intermediate hosts (including sheep, goats, rodents) to final hosts may induce the shedding of oocysts in these hosts and thus pose a risk for post-natal infection of cattle. With respect to oocyst morphology and the infectivity of muscle tissues for final hosts, no differences were seen in comparison with observations made in the past on Isospora bigemina/I. heydorni/H. heydorni. Therefore, earlier studies made on I. bigemina/I. heydorni/H. heydorni have to be re-evaluated critically to determine whether they may have included N. caninum or other protozoan parasites that use dogs as final hosts and have an oocyst morphology resembling that of I. bigemina/I. heydorni/H. heydorni.

Animals↗

Effect of antiretroviral therapy on cryptosporidiosis and microsporidiosis in patients infected with human immunodeficiency virus type 1.

To better understand whether potent antiretroviral therapies can modify the natural history of HIV-1-associated microsporidiosis and cryptosporidiosis, the response to antimicrobial treatment of these opportunistic infections was evaluated in patients with or without antiretroviral treatment. Fifty patients with diarrhoea, all positive for Cryptosporidium parvum or Enterocytozoon bieneusi, were included in the study. Retrospective data were collected concerning demographics, clinical and microbiological characteristics of the parasitic infection, antiretroviral therapy and prophylaxis against opportunistic infections. Faecal samples were prepared using the Richie formalin-ethyl acetate method and stained using the modified Ziehl-Neelsen method for detection of Cryptosporidium parvum and Isospora belli, the modified trichrome and calcofluor white technique for detection of Enterocytozoon spp., and iodine for detection of ova, cysts or vegetative forms. Diarrhoea was defined as an abnormal increase in stool liquidity, an abnormal increase in stool frequency and a daily stool weight of more than 250 g for a period of at least 4 days. Patients treated with double antiretroviral therapy or protease inhibitors demonstrated an excellent response and a sustained therapeutic effect after follow-up (range, 5-36 months). The relapse of cryptosporidiosis in two patients who discontinued antiretroviral therapy suggests that the infection might remain in a latent stage. The resolution of the diarrhoea seems to be related to an increased CD4+ cell count rather than to the viral load. In conclusion, these data strongly support the hypothesis that combination antiretroviral therapy is able to greatly modify the course of cryptosporidiosis and microsporidiosis in patients infected with HIV-1.

AIDS-Related Opportunistic Infections↗