Intestinal protozoa and epithelial cell kinetics, structure and function

Intestinal protozoa are not only common enteric pathogens in the tropics but also the high incidence of infection among immunocompromised patients in northern countries has evoked an increased interest in these parasites. Although enteric protozoa are a major cause of diarrhea and malabsorption in humans and other animals, the pathophysiology of gut disturbances caused by them remains poorly understood. Clinical signs related to enteric protozoan disease commonly involve malabsorption, diarrhea, weight loss or retarded weight gain and anorexua. Since these infections are most prevalent and most severe in the young, this may translate into considerable illness among children and significant loss to the agricultural economy where domestic animals are prone to infection. In this review we describe the effects of intestinal protozoan diseases on the structure, kinetics and function of absorptive intestinal cells and other epithelial cells, and correlate morphological injury with physiological alterations in the parasitized gut. Some of the interactions between immune responses and pathophysiology will be discussed, but in-depth discussion of intestinal immunity has recently been undertaken by other authors.     

Parasitism in Kansas in the 1800s: a glimpse to the past through the analysis of grave sediments from Meadowlark cemetery

During the excavations of the XIX century Meadowlark cemetery (Manhattan, Kansas, US), samples of sediments were taken from around five skeletons, and analyzed to detect intestinal parasites. No helminth eggs were found, but immunological ELISA tests for Entamoeba histolytica were positive in three samples. The immunological techniques have been successfully used in paleoparasitology to detect protozoan infections. Amoebiasis could have been a severe disease in the past, especially where poor sanitary conditions prevailed, and there is evidence that this cemetery may have been used in a situation where poor sanitary conditions may have prevailed. The presence of this protozoan in US during the late XIX century gives information on the health of the population and provides additional data on the parasite's evolution since its appearance in the New World.     

Intestinal Protozoa of the Baboon Papio doguera Pucheran, 1856

SYNOPSIS. Broad scope investigations have allowed a study of the intestinal parasitic protozoa of the Kenya baboon Papio doguera in its natural habitat as well as an opportunity to follow the intestinal protozoan populations in these primates held in captivity. Samplings indicate that the amoebae found in the baboon are essentially the same as those which commonly occur in man. Balantidium coli is frequent in wild baboons but is self-limiting in animals after a few weeks in captivity. The flagellates Chilomastix mesnili and Giardia lamblia were detected in captive but not in wild baboons; the presence of the latter constitutes a new record for Papio doguera.     

Cyclospora cayetanensis: a review of an emerging parasitic coccidian

Cyclospora cayetanensis is a sporulating parasitic protozoan that infects the upper small intestinal tract. It has been identified as both a food and waterborne pathogen endemic in many developing countries. It is an important agent of Traveller's Diarrohea in developed countries and was responsible for numerous foodborne outbreaks in the United States and Canada in the late 1990s. Like Cryptosporidium, infection has been associated with a variety of sequelae such as Guillain-Barré syndrome, reactive arthritis syndrome (formally Reiter syndrome) and acalculous cholecystitis.There has been much debate as to where to place C. cayetanensis taxonomically due to its homology with Eimeria species. To date, the only genomic DNA sequences available are the ribosomal DNA of C. cayetanensis and three other species; within these a high degree of homology has been observed. This homology and the lack of sequence data from other Cyclospora species have hindered identification methods.     

Short hairpin RNA-mediated knockdown of protein expression in Entamoeba histolytica

Background  

Entamoeba histolytica is an intestinal protozoan parasite of humans. The genome has been sequenced, but the study of individual gene products has been hampered by the lack of the ability to generate gene knockouts. We chose to test the use of RNA interference to knock down gene expression in Entamoeba histolytica.     

Giardia disrupts the arrangement of tight, adherens and desmosomal junction proteins of intestinal cells

Giardia duodenalis is a parasitic protozoan that causes diarrhea and other symptoms which together constitute a disease known as giardiasis. Although the disease has been well defined, the mechanisms involving the establishment of the infection have not yet been fully elucidated. In this study, we show that after 24h of interaction between parasites and intestinal Caco-2 cells, there was an alteration of the paracellular permeability, as observed by an approximate 42% of reduction in the transepithelial electrical resistance and permeation to ruthenium red, which was concomitant with ultrastructural changes. Nevertheless, epithelium viability was not affected. We also demonstrate that there was no change in expression of junctional proteins (tight and adherens) but that the distribution of these proteins in Caco-2 cells after parasite adhesion was significantly altered, as observed via laser scanning confocal microscopy 3D reconstruction. The present work shows that adhesion of Giardia duodenalis trophozoites to intestinal cells in vitro induces disturbances of the tight, adherens and desmosomal junctions.     

Prevalence and Risk Factors for Intestinal Protozoan Infections with Cryptosporidium,Giardia, Blastocystis and Dientamoeba among Schoolchildren in Tripoli,Lebanon

BackgroundIntestinal protozoan infections are confirmed as major causes of diarrhea, particularly in children, and represent a significant, but often neglected, threat to public health. No recent data were available in Lebanon concerning the molecular epidemiology of protozoan infections in children, a vulnerable population at high risk of infection.ConclusionsThis is the first study performed in Lebanon reporting the prevalence and the clinical and molecular epidemiological data associated with intestinal protozoan infections among schoolchildren in Tripoli. A high prevalence of protozoan parasites was found, with Blastocystis spp. being the most predominant protozoans. Although only 50% of children reported digestive symptoms, asymptomatic infection was observed, and these children may act as unidentified carriers. This survey provides necessary information for designing prevention and control strategies to reduce the burden of these protozoan infections, especially in children.     

Genomes and genome projects of protozoan parasites

Protozoan parasites are causing some of the most devastating diseases world-wide. It has now been recognised that a major effort is needed to be able to control or eliminate these diseases. Genome projects for the most important protozoan parasites have been initiated in the hope that the read-out of these projects will help to understand the biology of the parasites and identify new targets for urgently needed drugs. Here, I will review the current status of protozoan parasite genome projects, present findings obtained as a result of the availability of genomic data and discuss the potential impact of genome information on disease control.     

Oral dosing of neonatal mice with sucrose reduces infection with Cryptosporidium parvum

Cryptosporidium parvum is a significant cause of diarrheal disease in humans and economically important livestock species. There is no effective treatment available for this protozoan parasite. Mechanisms of intestinal colonization by C. parvum are not well understood, but it has been suggested that the parasite may utilize a lectin-like receptor. We used an infant mouse model to test whether high sugar concentrations in the intestine would affect in vivo colonization with C. parvum. We found that a single oral dose of sucrose, administered to mice at the time of, or 24 hr before, challenge with C. parvum significantly reduced infection. Significant reduction of infection was also seen in mice given isomaltose. Histologic examination of intestinal sections of mice treated with sucrose or isomaltose, but not other sugars, showed marked vacuolation of the small intestinal epithelium 1 day after treatment. Three days after treatment, tissue appeared normal. Thus, sucrose and, to a lesser extent, isomaltose reduced in vivo colonization with C. parvum and altered epithelial cell morphology in intestines of mice.     

The costs of dominance: testosterone, cortisol and intestinal parasites in wild male chimpanzees

Background

Male members of primate species that form multi-male groups typically invest considerable effort into attaining and maintaining high dominance rank. Aggressive behaviors are frequently employed to acquire and maintain dominance status, and testosterone has been considered the quintessential physiological moderator of such behaviors. Testosterone can alter both neurological and musculoskeletal functions that may potentiate pre-existing patterns of aggression. However, elevated testosterone levels impose several costs, including increased metabolic rates and immunosuppression. Cortisol also limits immune and reproductive functions.

Methods

To improve understanding of the relationships between dominance rank, hormones and infection status in nonhuman primates, we collected and analyzed 67 fecal samples from 22 wild adult male chimpanzees (Pan troglodytes schweinfurthii) at Ngogo, Kibale National Park, Uganda. Samples were analyzed for cortisol and testosterone levels as well as intestinal parasite prevalence and richness. 1,700 hours of observation data were used to determine dominance rank of each animal. We hypothesized that dominance rank would be directly associated with fecal testosterone and cortisol levels and intestinal parasite burden.

Results

Fecal testosterone (but not cortisol) levels were directly associated with dominance rank, and both testosterone and cortisol were directly associated with intestinal parasite richness (number of unique species recovered). Dominance rank was directly associated with helminth (but not protozoan) parasite richness, so that high ranking animals had higher testosterone levels and greater helminth burden.

Conclusions

One preliminary interpretation is that the antagonist pleiotropic effects of androgens and glucocorticoids place a cost on attaining and maintaining high dominance rank in this species. Because of the costs associated with elevated steroid levels, dominance status may be an honest signal of survivorship against helminth parasites.     

The intestinal parasite infection status of inhabitants in the Roxas city,The Philippines

We carried out a small-scale survey to investigate the status of intestinal protozoa and helminthes infection of inhabitants in Roxas city, Mindoro, the Philippines. Total 301 stool samples were subjected to the formalin-ether concentration method for the detection of helminth ova and protozoan cysts. The overall positive rate was 64.5%, and that of male and female were 56.6% and 72.5%, respectively. The highest infected helminth was Ascaris lumbricoudes (51.2%), followed by Trichuris trichiura (27.6%), hookworm (8.0%) and Enterobius vermicularis (0.3%). The protozoa infection status revealed that Entamoeba coli was the most frequent (15.0%). Iodoamoeba buetschlii and E. histolytica were found but few. The multiple infection more than two parasites was 29.6%, and double infection with A. lumbricoides and T. trichiura was most common. The intestinal helminth infections were highly prevalent in this area, according to this result, and we concluded that anthelminthic drugs should be given to inhabitants, especially to children of 1 to 15 years-old.     

Generation Time and Growth Rate of the Human Intestinal Parasite Blastocystis hominis

ABSTRACT. Blastocystis hominis , an anaerobic intestinal protozoan parasite of man, has a generation time (GT) in axenic culture of 8.5–19.4 h, depending on the strain tested. Average GT of the eight strains was 11.7 h. Zero growth time cell counts of 5.0 × 10⁵/ml to 2.0 × 10⁶/ml rose in 3–5 days to 1 × 10⁷ or 1 × 10⁸ cells/ml. The GT was determined for the 24-h period during which the most rapid growth occurred; about 2% of the B. hominis cells were in division during this time. Division under the culture conditions provided was by binary fission, the usual mode for B. hominis in vitro as well as in vivo. Division times were determined also by direct observation of individual dividing cells in slide cultures. These were usually ca. 40–60 min but sometimes as low as 20 min.     

雄性短尾猴的等级顺位、睾酮水平与肠道寄生虫感染强度的关系

在等级森严的非人灵长类群体中,睾酮对雄性顺位维持有着重要作用,且影响着体内寄生虫种类和荷虫量。本研究以栖息于安徽黄山的雄性短尾猴为研究对象,通过计算成年雄性的等级顺位,测定其粪便中睾酮含量以及寄生虫的荷虫量,探讨雄性等级顺位、睾酮水平与健康间的平衡关系。结果表明：共检测出10种肠道寄生虫,包括4种蠕虫、5种原虫和1种螨虫;其中雄性短尾猴个体间睾酮水平（1.00±0.42） ng/g和肠道寄生虫荷虫量（112.44±83.62）EPG均存在显著差异（Kruskal-Wallis H,P<0.05）;等级顺位与睾酮水平呈正相关（Spearman, N=8,ρ= 0.326, P<0.05）,与肠道原虫类的荷虫量呈负相关（Spearman, N=8,ρ= - 0.345, P<0.05）,与肠道蠕虫类的荷虫量无相关性（Spearman, N=8,ρ= 0.065, P>0.05）;睾酮水平与肠道原虫类寄生虫的荷虫量呈负相关（Spearman, N=8,ρ= -0.546, P<0.05）,与肠道蠕虫类的荷虫量无相关性（Spearman, N=8,ρ= -0.013, P>0.05）。本研究表明高顺位雄性拥有更高的睾酮水平,这不仅对其顺位的维持具有重要作用,对原虫类寄生虫也具有明显的抑制作用。     

Cytokinesis arrest and nuclear fission in low density populations of trichomonad protozoan

Cell growth of anaerobic protozoan Tritrichomonas foetus was analyzed. This protozoan usually proliferates in extremely high density, but protozoan parasites were dispersed uniformly in F-bouillon medium and cell division stopped temporarily. However, nuclear fission continued and giant polynucleated cells formed. Later, cell division resumed and cells returned to normal form. In conditioned medium, cytokinesis of the dispersed parasites did not stop. Results indicated that T. foetus cells secreted an extracellular factor that influenced cytokinesis.     

Entamoeba bubalus, n.sp., from Carabao

SUMMARY. Twelve carabao in the Philippines were found to be lightly infected with intestinal amoebae. Trophic forms (12 microns in diameter) of this protozoan possessed a definite ectoplasm and a homogeneous endoplasm. They were found only in stained preparations. The nucleus was similar to that of the cyst. All unconcentrated fecal smears contained at least a few cysts (8 microns in diameter). In these forms the cytoplasm usually contained a large vacuole and one or more irregular chromatoidal bodies. The nucleus (2.6 microns in diameter) possessed a pronounced, deeply staining, uniform peripheral ring and a large irregular endosome. There was no periendosomal ring. This amoeba is designated as Entamoeba bubalus , n.sp.     

A small-scale survey on the status of intestinal parasite infections in rural villages in Nepal

The status of intestinal parasite infections was investigated in two rural villages (Chitrasar, Jerona) in Chitwan District, Nepal in 1999. Stool examination was performed with a total of 300 specimens from schoolchildren by formalin-ether sedimentation technique. The prevalence rate of intestinal parasite infections in the surveyed areas was 44.0%. The prevalence rate in Jerona was slightly higher than that in Chitrasar. The prevalence rate of intestinal parasite infections in female was slightly higher than that in male without statistically significant difference. Entamoeba coli was the most commonly found protozoan parasite (21.0%) followed by Giardia lamblia (13.7%) and others (5.3%). Hookworm was the most prevalent intestinal helminth (13.0%) followed by Trichuris trichiura (3.0%) and others (5.0%). Forty-three specimens (14.3%) showed mixed infections. It is necessary to implement large-scale treatment with anthelminthics, health education and sanitary improvement for intestinal parasite control in the surveyed areas.     

Heat shock protein 90 from neglected protozoan parasites

Significant advances have been made in our understanding of heat shock protein 90 (Hsp90) in terms of its structure, biochemical characteristics, post-translational modifications, interactomes, regulation and functions. In addition to yeast as a model several new systems have now been examined including flies, worms, plants as well as mammalian cells. This review discusses themes emerging out of studies reported on Hsp90 from infectious disease causing protozoa. A common theme of sensing and responding to host cell microenvironment emerges out of analysis of Hsp90 in Malaria, Trypanosmiasis as well as Leishmaniasis. In addition to their functional roles, the potential of Hsp90 from these infectious disease causing organisms to serve as drug targets and the current status of this drug development endeavor are discussed. Finally, a unique and the only known example of a split Hsp90 gene from another disease causing protozoan Giardia lamblia and its evolutionary significance are discussed. Clearly studies on Hsp90 from protozoan parasites promise to reveal important new paradigms in Hsp90 biology while exploring its potential as an anti-infective drug target. This article is part of a Special Issue entitled: Heat Shock Protein 90 (HSP90).     

Synergic effect of metronidazole and pyrantel pamoate on Giardia lamblia

Giardia lamblia is a pathogenic protozoan presenting as the main characteristic, the trophozoite capacity to adhere in host intestinal epithelium, infecting mammals, including humans. The clinical treatment of this disease is based on metronidazole (Mz) that acts as an alternative electron acceptor, and its reduction promotes DNA impairment. In veterinary treatment, one of the best options is pyrantel pamoate (Pm), which the mode of action has not elucidated yet. Different strategies for Giardia treatment have been explored to avoid side effects to the host. In this context, the efficiency of treatment combining drugs raise as an interesting alternative for protozoan diseases. Here, we evaluated in vitro synergic effect of Mz and Pm on trophozoites and on its adherence to IEC-6 cells. The treatment with Mz or Pm was effective on trophozoites, with IC(50)/24h values of 5.3±0.9 μM and 13.8±1.4 μM, respectively. The treatment of trophozoites with different combinations of Mz and Pm were also evaluated, as showed by fractional inhibitory concentration index (FICI) under 0.5 in all conditions tested, corresponding to the synergic effect. This synergic activity was also observed when the combinations of 5.3 μM Mz+0.4 μM Pm and 13.8 μM Pm+0.1 μM Mz induced a remarkable reduction in % adhesion (85-90% and 52-59%, respectively) and in number of adhered parasites per 100 cells. The low cytotoxicity to the host cells of the combinations, associated to the strong synergic potential of the combination, encourage us to further investigate its effect in in vivo models.     

Present Status of Subclinical Intestinal Malabsorption in the Tropics

A review of the literature on subclinical intestinal disease shows its probable relevance in certain nutritional deficiencies in the indigenous residents of tropical climates. A significant incidence of intestinal disease has been defined among asymptomatic fit-looking adult Nigerians. The results of subsequent investigation into the pathogenesis of this disorder strongly point to hypoalbuminaemia as the important factor capable of producing a biochemically and histologically defined malabsorptive state leading to subnormal nutritional status among members of this community.