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

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

Blastocystis sp. is a common zoonotic intestinal protozoa which has been classified into 17 subtypes (STs). A cross-sectional study was conducted to determine the prevalence and subtype distribution of Blastocystis in villagers living on the Thai-Myanmar border, where the risk of parasitic infection is high. A total of 207 stool samples were collected and DNA was extracted. PCR and sequencing using primers targeting small-subunit ribosomal RNA (SSU rRNA) gene were performed. The prevalence of Blastocystis infection was 37.2% (77/207). ST3 (19.8%; 41/207) was the predominant subtype, followed by ST1 (11.6%; 24/207), ST2 (5.3%; 11/207), and ST4 (0.5%; 1/207). A phylogenetic tree was reconstructed using the maximum likelihood (ML) method based on the Hasegawa-Kishino-Yano + G + I model. The percentage of bootstrapped trees in which the associated taxa clustered together was relatively high. Some sequences of Blastocystis positive samples (TK18, 39, 46, 71, and 90) were closely related to animals (pig and cattle) indicating zoonotic risks. Therefore, proper health education in parasitic prevention for the villagers should be promoted to improve their personal hygiene. Further longitudinal studies are required to monitor the prevalence of parasitic infections after providing health education and to investigate Blastocystis ST in animals living in these villages.
Adult ; Aged ; Animals ; Blastocystis/*classification/immunology/*isolation & purification ; Blastocystis Infections/*parasitology ; Cluster Analysis ; Cross-Sectional Studies ; DNA, Protozoan/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Female ; Humans ; Male ; Middle Aged ; Myanmar ; Phylogeny ; RNA, Ribosomal, 18S/genetics ; Rural Population ; Sequence Analysis, DNA ; Seroepidemiologic Studies ; *Serogroup ; Thailand ; Young Adult

Microsporidia are eukaryotic organisms that cause zoonosis and are major opportunistic pathogens in HIV-positive patients. However, there is increasing evidence that these organisms can also cause gastrointestinal and ocular infections in immunocompetent individuals. In Korea, there have been no reports on human infections with microsporidia to date. In the present study, we used real-time PCR and nucleotide sequencing to detect Encephalitozoon intestinalis infection in seven of 139 human diarrheal stool specimens (5%) and Encephalitozoon hellem in three of 34 farm soil samples (8.8%). Genotype analysis of the E. hellem isolates based on the internal transcribed spacer 1 and polar tube protein genes showed that all isolates were genotype 1B. To our knowledge, this is the first report on human E. intestinalis infection in Korea and the first report revealing farm soil samples as a source of E. hellem infection. Because microsporidia are an important public health issue, further large-scale epidemiological studies are warranted.
AIDS-Related Opportunistic Infections/parasitology ; Adolescent ; Adult ; Aged ; Agriculture ; Base Sequence ; Child ; Child, Preschool ; DNA, Intergenic/genetics ; DNA, Protozoan/genetics ; Encephalitozoon/*genetics/*isolation & purification ; Encephalitozoonosis/*epidemiology ; Feces/*parasitology ; Female ; Fungal Proteins/genetics ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Molecular Typing ; Real-Time Polymerase Chain Reaction ; Republic of Korea/epidemiology ; Sequence Alignment ; Sequence Analysis, DNA ; Soil/*parasitology ; Young Adult

No abstract available.
Female ; Humans ; Lung Diseases, Parasitic/*parasitology ; Parabasalidea/*isolation & purification ; Protozoan Infections/*parasitology

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

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

Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.
Animals ; DNA, Protozoan/genetics ; DNA, Ribosomal Spacer/genetics ; Humans ; Indonesia/epidemiology ; *Mammals ; Protozoan Infections/epidemiology/*parasitology ; RNA, Protozoan/genetics ; RNA, Ribosomal, 18S/genetics ; Species Specificity ; Trichomonadida/*classification/*genetics/isolation & purification

This study aimed to measure the levels of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin 1 (IL-1), interleukin 6 (IL-6), and nitrite/nitrate (NOx) in serum of dogs experimentally infected with Rangelia vitalii. Twelve female mongrel dogs were divided into 2 groups; group A (uninfected controls) composed by healthy dogs (n=5) and group B consisting of dogs inoculated with R. vitalii (n=7). Animals were monitored by blood smear examinations, which showed intraerythrocytic forms of the parasite on day 5 post-infection (PI). Blood samples were collected through the jugular vein on days 0, 10, and 20 PI to determine the serum levels of IFN-gamma, TNF-alpha, IL-1, IL-6, and NOx. Cytokines were assessed by ELISA quantitative sandwich technique, and NOx was measured by the modified Griess method. Cytokine levels (IFN-gamma, TNF-alpha, IL-1, and IL-6) were increased (P<0.01) in serum of infected animals. Serum levels of NOx were also increased on days 10 PI (P<0.01) and 20 PI (P<0.05) in infected animals. Therefore, the infection with R. vitalii causes an increase in proinflammatory cytokines and nitric oxide content. These alterations may be associated with host immune protection against the parasite.
Animals ; Chemistry Techniques, Analytical ; Cytokines/*blood ; Disease Models, Animal ; Dogs ; Enzyme-Linked Immunosorbent Assay ; Female ; Nitric Oxide/*blood ; Piroplasmida/*immunology ; Protozoan Infections/*immunology/parasitology/pathology ; Serum/chemistry

After bathing at a hot spring resort, a 75-year-old man presented to the emergency department because of seizure-like attack with loss of conscious. This is the first case of primary amebic meningoencephalitis (PAM) caused by Naegleria fowleri in Taiwan. PAM was diagnosed based on detection of actively motile trophozoites in cerebrospinal fluid using a wet-mount smear and the Liu's stain. The amoebae were further confirmed by PCR and gene sequencing. In spite of administering amphotericin B treatment, the patient died 25 days later.
Aged ; Amebiasis/*diagnosis/parasitology/*pathology ; Central Nervous System Protozoal Infections/*diagnosis/parasitology/*pathology ; Cerebrospinal Fluid/parasitology ; DNA, Protozoan/chemistry/genetics ; Fatal Outcome ; Humans ; Male ; Microscopy ; Naegleria fowleri/classification/genetics/*isolation & purification ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Taiwan