• Title/Summary/Keyword: protozoan

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DNA Extraction from Protozoan Oocysts/Cysts in Feces for Diagnostic PCR

  • Hawash, Yousry
    • Parasites, Hosts and Diseases
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    • v.52 no.3
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    • pp.263-271
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    • 2014
  • 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 ($50-100{\mu}l$) 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, ${\approx}2$ 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.

Growth and Succession of Protozoa Population in the Activated Sludge Plant. (하수처리장의 원생동물 천이)

  • 이찬형;문경숙;진익렬
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.158-165
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    • 2004
  • This study concerns the succession process of protozoa populations in two activated sludge pilot plants. Protozoa populations with similar structures developed and succeeded each other in two plants. It was observed that initial sludge inoculum did not influence the protozoan composition but shorten the period of the plant stabilization. It seemed the protozoan in aeration tank was come from the influent. The composition of the influent sewage influenced the structure of protozoa in the pilot plants. The flagellates was very abundant during the initial starting phase. These rapidly decreased after growth peak. Then sarcodina, ciliates were increased in succession. In ciliates, free-swimming forms appeared at first and crawling forms and stalked forms were followed. After 36 days from the start, the sludge stabilization phase was characterized by a populations of ciliates dominated by stalked and crawling type and low value of effluent BOD was achieved.

A small-scale survey of intestinal parasite infections among children and adolescents in Legaspi city, the Philippines

  • Lee, Kyu-Jae;Ahn, Yung-Kyum;Yong, Tai-Soon
    • Parasites, Hosts and Diseases
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    • v.38 no.3
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    • pp.183-185
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    • 2000
  • To determine the status of infection caused by intestinal parasites among children and adolescents living in Legaspi city, the Philippines, we performed a small survey by fecal examination for helminth ova and protozoan cysts with formalin-ether concentration method. Of the 64 examinees, the infection rate was 78.1%. The infection rates of primary school children, preschool children and adolescents were 95.5%, 64.7% and 87.5%, respectively. The infection rate in urban areas was 56%, and 92.3% in rural areas. The infection rates were 51% with Trichuris trichiura, 40% with Ascaris lumbricoides, 23.4% with hookworm, 15.6% with Iodamoeba butschlii, 14.1% with Endolinax nana, 9.4% with Entamoeba coli and 7.8% with Giardia lamblia. There were 33 cases with multiple infection (51.6%). Mixed infection with more than 3 parasites was observed in 15 cases, all of them being children and adolescents living in rural areas. By this survey, it was conjectured that helminthic infection is prevalent among children and adolescents in Legaspi, Philippines. To determine the status of infection caused by intestinal parasites among children and adolescents living in Legaspi city, the Philippines, we performed a small survey by fecal examination for helminth ova and protozoan cysts with formalin-ether concentration method. Of the 64 examinees, the infection rate was 78.1%. The infection rates of primary school children, preschool children and adolescents were 95.5%, 64.7% and 87.5%, respectively. The infection rate in urban areas was 56%, and 92.3% in rural areas. The infection rates were 51% with Trichuris trichiura, 40% with Ascaris lumbricoides, 23.4% with hookworm, 15.6% with Iodamoeba butschlii, 14.1% with Endolinax nana, 9.4% with Entamoeba coli and 7.8% with Giardia lamblia. There were 33 cases with multiple infection (51.6%). Mixed infection with more than 3 parasites was observed in 15 cases, all of them being children and adolescents living in rural areas. By this survey, it was conjectured that helminthic infection is prevalent among children and adolescents in Legaspi, Philippines.

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Temporal and Spatial Distribution of Biomass and Cell Size of Bacteria and Protozoa in Lake Paldang and Kyungan Stream (팔당호와 경안천에서 박테리아와 원생생물의 생물량과 세포크기의 시 ${\cdot}$ 공간적 분포)

  • Son, Ju-Youn;Kong, Dong-Soo;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.39 no.3 s.117
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    • pp.378-389
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    • 2006
  • Seasonal changes of biomass and cell size of bacteria and protozoa, and factors affecting their distribution in Lake Paldang and Kyungan Stream were analyzed from April to December, 2005. Bacterial abundance at Paldang Dam and Kyungan Stream was similar, but it did not much increase during hot summer period. Protozoan carbon biomass was much greater at Kyungan Stream compared to Paldang Dam. HNAN generally accounted for the majority of total protozoan biomass, but ciliates made up the highest proportion in April and November at Paldang Dam and June at both sites. PNAN showed low biomass at both sites, but it was high during spring and fall season. Small-sized HNAN ($3{\sim}7\;{\mu}m$) numerically predominated the protozoan community at both sites. Average cell size of HNAN was bigger at Kyungan Stream where nutrients concentration was much higher than Paldang Dam. Average cell size of ciliates varied seasonally; it was relatively small during the summer. HNAN biomass significantly correlated with Chl-a concentration and ciliates biomass at Paldang Dam, indicating that HNAN increase might link to the ciliates increase. At Kyungan Stream, HNAN biomass showed a significant relationship with PNAN biomass, and Chl-a concentration was closely related with both of HNAN and PNAN biomass. Ciliate biomass showed significant relationship with nutrient (TN, TP) and particulate matter (SS) only at Kyungan Stream. At both sites, protozoan biomass was significantly correlated with bacterial biomass, and ciliates were additionally related flagellates. High biomass of microbial components and the close relationships among them suggest that the energy transfer through the microbial loop may important in the plankton food web of Lake Paldang ecosystem.

Influence of 120 kDa Pyruvate:Ferredoxin Oxidoreductase on Pathogenicity of Trichomonas vaginalis

  • Song, Hyun-Ouk
    • Parasites, Hosts and Diseases
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    • v.54 no.1
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    • pp.71-74
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    • 2016
  • Trichomonas vaginalis is a flagellate protozoan parasite and commonly infected the lower genital tract in women and men. Iron is a known nutrient for growth of various pathogens, and also reported to be involved in establishment of trichomoniasis. However, the exact mechanism was not clarified. In this study, the author investigated whether the 120 kDa protein of T. vaginalis may be involved in pathogenicity of trichomonads. Antibodies against 120 kDa protein of T. vaginalis, which was identified as pyruvate:ferredoxin oxidoreductase (PFOR) by peptide analysis of MALDI-TOF-MS, were prepared in rabbits. Pretreatment of T. vaginalis with anti-120 kDa Ab decreased the proliferation and adherence to vaginal epithelial cells (MS74) of T. vaginalis. Subcutaneous tissue abscess in anti-120 kDa Ab-treated T. vaginalis-injected mice was smaller in size than that of untreated T. vaginalis-infected mice. Collectively, the 120 kDa protein expressed by iron may be involved in proliferation, adhesion to host cells, and abscess formation, thereby may influence on the pathogenicity of T. vaginalis.

Trypanosome Glycosylphosphatidylinositol Biosynthesis

  • Hong, Yeon-Chul;Kinoshita, Taroh
    • Parasites, Hosts and Diseases
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    • v.47 no.3
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    • pp.197-204
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    • 2009
  • Trypanosoma brucei, a protozoan parasite, causes sleeping sickness in humans and Nagana disease in domestic animals in central Africa. The trypanosome surface is extensively covered by glycosylphosphatidylinositol (GPI)-anchored proteins known as variant surface glycoproteins and procyclins. GPI anchoring is suggested to be important for trypanosome survival and establishment of infection. Trypanosomes are not only pathogenically important, but also constitute a useful model for elucidating the GPI biosynthesis pathway. This review focuses on the trypanosome GPI biosynthesis pathway. Studies on GPI that will be described indicate the potential for the design of drugs that specifically inhibit trypanosome GPI biosynthesis.

Evaluation of the membrane filtration and elution for detection of Giardia lamblia cysts in water (수중 Giardia lamblia cysts 검출을 위한 멤브레인 여과 및 용출에 대한 연구)

  • Hong, Uk-Seon;Kim, Gyeong-Ju;Lee, Gi-Se
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.422-423
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    • 2000
  • The protozoan parasite Giardia lamblia has been implicated as the causative agents of many outbreaks of waterborne intestinal illness. Accurate evaluation of Giardia lamblia removal in water treatment process requires a reliable method for measuring the concentrations of these pathogens in water. The relative recovery of Giardia cysts was assessed for seeded samples of distilled water. Cysts preparation was done by encystment in vitro. Membrane filtration was evaulated with cellulose acetate, polycarbonate, polypropylene, polyethersulfone, nylon membranes. Elution conditions were varied to improve cyst recovery.

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Histopathological changes of small intestine of cats infected with helminth (연충감염 고양이 소장의 병리조직학적 변화)

  • 양홍지;박태욱;김현철;김태중;박배근
    • Korean Journal of Veterinary Service
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    • v.19 no.3
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    • pp.199-205
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    • 1996
  • Helminth and their ova or protozoan oocysts from fecal samples of cats in Chonju and Daejon were examined from December 1995 to January 1996. Out of the 43 samples, 41(95.35%) were positive and 66(153.49%) were cumulative positive for parasite infection. The identified parasites were summerized as follows; Worms of Toxocara cati(62.79%), Taenia taeniformis(48.88%), Spirometra sp(23.25%) and Diphyllobothrium latum (6.97%), eggs of Clonorchis sinensis(2.32%), Ancylostoma tubaeformae(2.32%) and Capillaria sp(4.65%), oocyst of Isospora (2.32%) sp. In haevy infected intestine, mast cells(38 cells/10 villi) were detected in villi and goblet cells(131 cells/$0.1mm^2$) were severely increased in mucosa. Epithelium of intestine was partially deteched B. The end of villi were necrosed and hyperaemia was widely recognized in lamina propria. T taeniaeformis have not the neck, but have 2 layed hooks(anterior : 19, posterior : 19, total : 38).

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Cloning and Expression of a Serine Proteinase Gene Fragment from Acanthamoeba culbertsoni

  • Park, Ki-Won;Kim, Tong-Soo;Na, Byoung-Kuk;Song, Chul-Yong
    • BMB Reports
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    • v.31 no.3
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    • pp.303-306
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    • 1998
  • Serine proteinase cDNA fragment from protozoan parasite Acanthamoeba culbertsoni was amplified by the reverse transcription-polymerase chain reaction (RTPCR) using degenerate oligonucleotide primers derived from conserved serine proteinase sequences. The amplified DNA fragment was subcloned and sequenced. The sequence analysis and alignment showed significant sequence similarity to other eukaryotic serine proteinases and conservation of the His, Asp, and Ser residues that form the catalytic triad. The cDNA fragment was cloned into the pGEMEX-1 expression vector and expressed in Escherichia coli. A resulting fusion protein of 56 kDa had proteolytic activity. The fusion protein reacted with sera of mice immunized with purified serine proteinase of A. culbertsoni in Western blot. Immune recognition of the fusion protein by mouse antisera suggested that the fusion protein may be valuable as a diagnostic reagent.

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Balantidiasis in Gastric Lymph Node of Barbary Sheep (Ammotragus lervia)

  • Park, Nam-yong;Cho, Ho-seong;A.W.M. Effendy;Park, Jong-woog;Kim, Tae-soon;Shin, Sung-shik
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2003.10a
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    • pp.39-39
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    • 2003
  • Balantidiasis is an infectious disease worldwide which is produced by a protozoan Balantidium coli. This single-celled organism is characterized by their large size ranging from 50 $\mu\textrm{m}$ to more than 500 $\mu\textrm{m}$ [1] which indicated by the presence of cilia on its cell surface. The parasite occurs in the lumen of cecum and colon of swine, humans and nonhuman primates as commensal, but can turn opportunist and invade injury tissues by other diseases [2]. It is difficult to diagnose the disease clinically since they are asymptomatic [3]; and can be complicated with other disease or parasitism. Here we report the incidental findings of Balantidiasis in the lymphatic ducts of gastric lymph node of Barbary sheep (Ammotragus lervia). (omitted)

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