PCR detection of intestinal protozoa is often restrained by a poor DNA recovery or by inhibitors present in feces. The need for an extraction protocol that can overcome these obstacles is therefore clear. QIAamp(R) DNA Stool Mini Kit (Qiagen) was evaluated for its ability to recover DNA from oocysts/cysts directly from feces. Twenty-five Giardia-positive, 15 Cryptosporidium-positive, 15 Entamoeba histolytica-positive, and 45 protozoa-free samples were processed as control by microscopy and immunoassay tests. DNA extracts were amplified using 3 sets of published primers. Following the manufacturer's protocol, the kit showed sensitivity and specificity of 100% towards Giardia and Entamoeba. However, for Cryptosporidium, the sensitivity and specificity were 60% (9/15) and 100%, respectively. A series of optimization experiments involving various steps of the kit's protocol were conducted using Cryptosporidium-positive samples. The best DNA recoveries were gained by raising the lysis temperature to the boiling point for 10 min and the incubation time of the InhibitEX tablet to 5 min. Also, using a pre-cooled ethanol for nucleic acid precipitation and small elution volume (50-100 microl) were valuable. The sensitivity of the amended protocol to Cryptosporidium was raised to 100%. Cryptosporidium DNA was successfully amplified by either the first or the second primer set. When applied on parasite-free feces spiked with variable oocysts/cysts counts, approximately 2 oocysts/cysts were theoretically enough for detection by PCR. To conclude, the Qiagen kit with the amended protocol was proved to be suitable for protozoan DNA extraction directly from feces and support PCR diagnosis.
DNA, Protozoan/*isolation & purification ; Feces/*parasitology ; Humans ; Molecular Diagnostic Techniques/*methods ; Polymerase Chain Reaction/*methods ; Protozoan Infections/*diagnosis ; Sensitivity and Specificity ; Specimen Handling/*methods ; Spores, Protozoan/*genetics

Bronchopneumonia ; diagnosis ; parasitology ; Female ; Humans ; Lung Diseases, Parasitic ; diagnosis ; parasitology ; Male ; Protozoan Infections ; diagnosis ; parasitology

Human infections with Lophomonas blattarum are rare. However, the majority of the infections occurred in China, 94.4% (136 cases) of all cases in the world. This infection is difficult to differentiate from other pulmonary infections with similar symptoms. Here we reported a case of L. blattarum infection confirmed by bronchoalveolar lavage fluid smear on the microscopic observations. The patient was a 21-year-old female college student. The previous case which occurred in Chongqing was 20 years ago. We briefly reviewed on this infection reported in the world during the recent 20 years. The epidemiological characteristics, possible diagnostic basis, and treatment of this disease is discussed in order to provide a better understanding of recognition, diagnosis, and treatment of L. blattarum infection.
Female ; Humans ; Lung Diseases, Parasitic/diagnosis/*parasitology ; Parabasalidea/*isolation & purification ; Protozoan Infections/*parasitology ; Young Adult

Cell Movement ; Cilia ; physiology ; Epithelial Cells ; physiology ; ultrastructure ; Humans ; Hypermastigia ; ultrastructure ; Protozoan Infections ; diagnosis

Bronchopneumonia ; diagnosis ; parasitology ; Diagnosis, Differential ; Diagnostic Errors ; Humans ; Lung Diseases, Parasitic ; diagnosis ; parasitology ; Protozoan Infections ; diagnosis ; parasitology

Histomonas meleagridis is a facultative anaerobic parasite, which can cause a common poultry disease known as histomoniasis. The species and age of the birds impacts on the susceptibility, with turkey being the most susceptible species. Chickens are less susceptible to H. meleagridis than turkeys and usually serve as reservoir hosts. Here, the diagnosis of an outbreak of histomoniasis in backyard Sanhuang chickens is described. The primary diagnosis was made based on clinical symptoms, general changes at necropsy, histopathology, and the isolation and cultivation of parasites. The pathogen was further confirmed by cloning, PCR identification, and animal inoculation tests. A strain of H. meleagridis, named HM-JSYZ-C, with a higher pathogenicity level in chickens was obtained. The study lays a foundation for further investigations into H. meleagridis and histomoniasis in chickens.
Animals ; Birds ; Chickens ; Clone Cells ; Cloning, Organism ; Diagnosis ; Parasites ; Polymerase Chain Reaction ; Poultry Diseases ; Protozoan Infections ; Turkey ; Turkeys ; Virulence

In Korea, patterns of parasitic infections have notably changed during the past few decades. The soil-transmitted helminthiases and water-borne protozoan infections, which had been prevalent, became negligible, while parasitic zoonosis including pet-associated infections, food-borne helminthiases, and imported tropical endemic diseases have increasingly been detected. People who travel abroad and those who have immigrated from other countries might suffer from endemic tropical diseases. Except for a few entities, which invoked acute febrile illness (malaria) and diarrhea (giardiasis and amoebiasis), most helminthic and protozoan infections did not provoke acute symptoms. Those infections progress slowly, but can sometimes result in fatal clinical consequences. Diverse tropical endemic diseases are prevalent in several continents/countries according to different natural environments (climate and humidity), socioeconomic status, and traditional cultural background. Those diseases might be acquired through different routes of infection. Travelers who have returned to Korea from overseas and immigrants should undergo a careful differential diagnosis. Information on countries and duration of travel/residence, food habits, underlying medical history, prophylactics received, exposure to harmful environments (insect bites, contaminated food or water), and swimming in freshwater is valuable. This article briefly overviews the epidemiology, pathophysiology, clinical manifestations, diagnosis, and specific chemotherapeutics of the tropical endemic diseases that are important in Korea.
Diagnosis ; Diagnosis, Differential ; Diarrhea ; Emigrants and Immigrants ; Endemic Diseases ; Epidemiology ; Food Habits ; Fresh Water ; Helminthiasis ; Helminths ; Humans ; Korea ; Parasitic Diseases* ; Protozoan Infections ; Social Class ; Swimming

OBJECTIVES: To determine the prevalence and risk factors associated with intestinal parasites in the population of San Juan Cosala, Jalisco, Mexico. METHODS: A total of 277 samples from 104 participants were analysed using direct smear, flotation, formaldehyde/ethyl acetate, and modified Kinyoun’s acid-fast stain methods. The Graham method was applied only for samples from children under 12 years of age for the diagnosis of Enterobius vermicularis. RESULTS: The prevalence of parasite infections in the study population was 77.9% including: Entamoeba histolytica/E. dispar/E. moshkovskii/E. bangladeshi (37.5%), Giardia intestinalis (11.5%); commensals: Endolimax nana (44.2%), Entamoeba coli (27.9%), Chilomastix mesnili (6.7%) and Iodamoeba bütschlii, (2.9%); emerging intestinal protozoans: Blastocystis spp. (49%), Cryptosporidium spp. (7.7%) and Cyclospora cayetanensis (2.9%); and helminths: Enterobius vermicularis (18.3%) and Ascaris lumbricoides (5.8%). The results also showed that 58.64% of the studied population presented polyparasitism. A significant association was found between protozoan infections and housewives, and houses that were not built with concrete ceilings, brick walls and cement floors (p < 0.05). CONCLUSION: Polyparasitism was observed in over half the study population. The most prevalent parasite was Blastocystis spp, whilst the prevalence of helminths was less than that of protozoans. The risk factors for infection to intestinal parasites were being a housewife and not having solid brick, cement and concrete materials for house construction.
Adult ; Ascaris lumbricoides ; Blastocystis ; Child ; Cryptosporidium ; Cyclospora ; Diagnosis ; Endolimax ; Entamoeba ; Enterobius ; Giardia lamblia ; Helminths ; Humans ; Methods ; Mexico ; Parasites ; Prevalence ; Protozoan Infections ; Retortamonadidae ; Risk Factors

This study investigated the effect of breast-feeding in protection against protozoan infection in infants with persistent diarrhea. Infants were classified into 2 groups; 161 breast-fed infants and the same number of non-breast-fed infants. Microscopic examinations of stool were done for detection of parasites and measuring the intensity of infection. Moreover, serum levels of IgE and TNF-alpha were measured by ELISA. Cryptosporidium spp., Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Blastocystis sp. were demonstrated in infants with persistent diarrhea. The percentage of protozoan infections was significantly lower in breast-fed infants than that in the non-breast-fed infants. The levels of IgE and TNF-alpha were significantly lower in the breast-fed group than in the non-breast-fed group. There were significant positive associations between the serum levels of IgE and TNF-alpha and the intensity of parasite infection in the breast-fed group. It is suggested that breast-feeding has an attenuating effect on the rate and intensity of parasite infection.
Antigens, Protozoan/analysis/*immunology ; Diarrhea, Infantile/*diagnosis/parasitology ; Entamoeba ; Entamoeba histolytica/*isolation & purification ; Entamoebiasis/*diagnosis/parasitology ; Enzyme-Linked Immunosorbent Assay ; Feces/parasitology ; Female ; Giardia lamblia ; Giardiasis/*diagnosis/parasitology ; Humans ; Infant ; Intestines/parasitology ; Protozoan Infections/*diagnosis/parasitology ; Tumor Necrosis Factor-alpha/metabolism

OBJECTIVETo investigate the infection of Cryptosporidium and its epidemiological characteristics in AIDS patients of Southern China.

METHODSStool samples colleted from AIDS confirmed patients. The samples were detected for oocyst of Cryptosporidium by acid fast bacteria stain and indirect fluorescent antibody stain respectively, CD4 count was detected by Flow Cytometry.

RESULTS212 samples of fresh stool obtained from the AIDS patients who live in Guangdong and Yunnan province. The total infection rate of Cryptosporidium in AIDS patients was 4.25% (9/212), the infectious rate of oocyst in the group of 50- 59-years-old was significantly higher than those in 30-39 (P < 0.01); the infectious rate of oocyst in patients with antiretroviral therapy (ART) was also significantly lower (P = 0.0000); we found the patients coinfected with Cryptosporidium with CD4 count all below 100 cells/microl. However, there were no any difference between the infectious rate to the patient's gender, areas and stool shape.

CONCLUSIONAIDS patients infected by Cryptosporidium are not rare in southern China, and the infectious rate was lower than western country. Patients received ART could decrease the infectious rate of Cryptosporidium, Cryptosporidium always happen in patient whose CD4 count was very low (< 100 cells/microl).

AIDS-Related Opportunistic Infections ; parasitology ; Acquired Immunodeficiency Syndrome ; complications ; parasitology ; Animals ; Antigens, Protozoan ; CD4 Lymphocyte Count ; China ; Cryptosporidiosis ; diagnosis ; etiology ; immunology ; parasitology ; Cryptosporidium ; chemistry ; isolation & purification ; Feces ; parasitology ; Flow Cytometry ; HIV Infections ; parasitology ; Humans ; Oocysts ; Staining and Labeling