• Parasites, Hosts and Diseases
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• 제52권3호
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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.

• 한국미생물·생명공학회지
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• 제32권2호
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• pp.158-165
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• 2004

Pilot Plant 에서 도시하수가 처리되는 경우와 전 처리된 분뇨가 하수에 섞여 처리되는 두 가지 종류의 처리장하수를 대상으로 식종과 무식종의 경우로 나누어 활성슬러지의 정착 단계까지 조사하였다. 원생동물의 천이과정은 편모충류, 육질충류, 섬모충류 순이며, 섬모충류 내에서는 자유 유영형이 먼저 출현하고 포복형과 고착형이 차츰 증가하여 활성슬러지 원생동물상을 구성한다. 정상적으로 가동되는 처리장 포기조의 원생동물분포는 시운전초기단계의 원생동물 분포와 다르며, 시운전 안정화 단계에서의 원생동물 분포와 유사하였다. 포기조의 원생동물은 유입수에서 유래하는 것으로 판단되며, 포기조에 유입된 후 환경조건에 맞는 종이 증식하여 활성슬러지를 구성한다. 슬러지식종은 활성슬러지 원생동물 군집구조 형성에 영향을 미치지 못했으나 활성슬러지의 안정화기간을 다소 단축시키는 효과가 있었다. 처리장별로 활성슬러지 원생동물의 종류는 비슷하나 개체수는 달라 유입수의 성상에 영향을 받는 것으로 판단된다. Pilot plant 가동 후반기에는 섬모충류 비율이 70%전후, 개체수도 3,000∼5,000 개체/$m\ell$이며, 유출수의 BOD 농도도 안정적으로 나타나 정상적으로 가동된다고 판단된다.

• Parasites, Hosts and Diseases
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• 제38권3호
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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.

• 생태와환경
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• 제39권3호통권117호
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• pp.378-389
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• 2006

본 연구는 2005년 4월부터 12월까지 팔당댐과 경안천에서 박테리아와 원생생물 생물량과 개체크기의 계절적 변화와 이에 영향을 미치는 요인들에 대하여 분석하였다. 조사기간 동안 박테리아의 밀도는 팔당댐과 경안천에서 각각 $2.8{\sim}64.6\;{\times}\;10^6$, $6.1{\sim}43.3\;{\times}\;10^6\;cells\;{\cdot}\;mL^{-1}$로서 두 지점에서 비슷한 분포를 보였고, 수온이 높은 여름동안에도 증가하지 않았다. 원생생물의 총생물량은 팔당호 ($12{\sim}73\;{\mu}gC\;{\cdot}\;L^{-1}$)에 비해 경안천 ($26{\sim}169\;{\mu}gC\;{\cdot}\;L^{-1}$)에서 훨씬 높았으며, 평균 세포크기도 경안천에서 크게 나타났다. 원생생물의 총생물량 중에서 HNAN이 전반적으로 가장 큰 비중을 차지하였으나, 팔당댐에서 4, 10, 11월에 경안천에서 6월에는 섬모충류의 비중이 가장 컸다. PNAN은 상대적으로 매우 낮은 생물량을 보였으나 봄과 가을동안에는 높게 나타났다. HNAN은 두 지점에서 모두 3{\sim}7\;{\mu}m$ 크기의 작은 개체들이 우점하였다. HNAN 평균 세포의 크기는 영양염의 농도가 훨씬 높았던 경안천에서 더 크게 나타났다. 섬모충의 세포당 평균 크기는 여름 동안에 작았다. 팔당댐에서 HNAN 생물량은 Chl-a 농도와 상관성이 있었으며, 또한 섬모충류의 생물량과 관련이 높게 나타나 HNAN의 증가가 섬모충류의 높은 생물량으로 연결된 것으로 판단되었다. 경안천에서 PNAN과 HNAN은 Chl-a 농도와 높은 상관성을 보였고, PNAN의 생물량이 증가할수록 HNAN의 생물량도 증가하였다. 섬모충류의 생물량은 다른 생물보다는 영양염 (TN, TP)과 입자성 물질(SS)과 높은 상관성을 보였다. 또한 두 지점 모두에서 원생생물의 생물량은 박테리아 생물량과 밀접한 관계를 보였고, 섬모충류는 박테리아 뿐만 아니라 편모류의 생물량과도 밀접한 관계가 있는 것으로 나타났다. 팔당호 생태계에서 본 연구의 결과가 보여준 microbial loop 구성요인들의 높은 생물량과 이들 간의 밀접한 상관관계는 팔당호 생태계 플랑크톤 먹이망 내에서 microbial loop를 통한 먹이망 기능이 중요함을 시사한다.

• Parasites, Hosts and Diseases
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• 제54권1호
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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.

• Parasites, Hosts and Diseases
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• 제47권3호
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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.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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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.

• 한국동물위생학회지
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• 제19권3호
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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).

• BMB Reports
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• 제31권3호
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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.

• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2003년도 추계학술대회초록집
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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)