• Title/Summary/Keyword: Two-Color Method

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Trial for Drug Susceptibility Testing of Mycobacterium tuberculosis with Live and Dead Cell Differentiation (세포 염색 방법을 이용한 결핵균 감수성 검사법)

  • Ryu, Sung-Weon;Kim, Hyun-Ho;Bang, Mun-Nam;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Kang, Seongman;Bai, Gill-Han
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.3
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    • pp.261-268
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    • 2004
  • Background : The resurgence of tuberculosis and outbreaks of multidrug resistant (MDR) tuberculosis have increased the emphasis for the development of new susceptibility testing of the Mycobacterium tuberculosis for the effective treatment and control of the disease. Conventional drug susceptibility testings, such as those using egg-based or agar-based media have some limits, such as the time required and difficulties in determining critical inhibitory concentrations, but these are still being used in many diagnostic laboratories because of no better lternatives, considering cost and accuracy. To overcome these limits, a rapid and simple method for new susceptibility testing, using live and dead assays, was applied for a bacterial cell viability assay to distinguish dead from live bacterial cells based on two-color fluorescence. Materials and Methods Strains : Forty strains were used in this study, 20 susceptible to all antituberculosis drugs and the other 20 resistant to the four first line antituberculosis drugs isoniazid, rifampicin, streptomycin and ethambutol. Antibiotics : The four antibiotics were dissolved in 7H9 broth to make the following solutions: $0.1{\mu}g\;isoniazid(INH)/m{\ell}$, $0.4{\mu}g\;rifampicin(RMP)/m{\ell}$, $4.0{\mu}g\;streptomycin(SM)/m{\ell}$ and $4.0{\mu}g\;ethambutol(EMB)/m{\ell}$. Results : Live and dead Mycobacterium tuberculosis cells fluoresced green and red with the acridin (Syto 9) and propidium treatments, respectively. These results are very well accorded with conventional drug susceptibility testing by proportional method on Lowensen-Jensen media (L-J) containing 4 drugs (INH, RMP, EMB and SM), showing a 93.7 % accordance rate in susceptible strains and 95% in resistant strains. Conclusion : The results of the drug susceptibility testing using the live and dead bacterial cell assay showed high accordance rates compared with the conventional proportion method on L-J. This finding suggests that the live and dead bacterial cell assay can be used as an alternative to conventional drug susceptibility testing for M. tuberculosis strains.

Effects of quality grade, trimming, and packaging method on shelf life of king oyster mushrooms (큰느타리의 품질 등급, 손질 및 포장 방법에 따른 유통 수명)

  • Choi, Ji-Weon;Lee, Ji Hyun;Oh, In-Ho;Lim, Sooyeon;Im, Ji-Hoon;Yang, Hae Jo;Choi, Hyunjin;Shin, Sheob;Hong, Yoon Pyo
    • Journal of Mushroom
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    • v.19 no.3
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    • pp.234-245
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    • 2021
  • To extend the shelf life of king oyster mushrooms for export, we investigated the impacts of mushroom quality grade, fruiting body trimming, and packaging method (tray container sealed packaging vs string-tied OPP bag packaging). Quality is divided into two grades: 1st grade, which is mushrooms adapted by lowering the cultivation temperature to 9~11℃, and 2nd grade, mushrooms held at 13~15℃ prior to harvest. Using selected 1st and 2nd grade mushrooms, 3 treatments were carried out to assess effects of trimming and packaging method. Test groups included 1) trimming plus string-tied OPP bag packaging (Cut & OPP), 2) no trimming plus string-tied OPP bag packaging (Uncut & OPP), and 3) trimming plus tray container sealing packaging (Cut & Tray). Gas composition inside the packaging, changes in quality factors, and sensory evaluation for fresh quality were performed over 42 days of 0℃ storage. Overall freshness was best maintained in the following order: Cut & Tray > Cut & OPP > Uncut & OPP for both 1st and 2nd grade mushrooms. The shelf-life of 1st grade mushrooms was about 30 days for Cut & Tray, 28 days for Cut & OPP, and 21 days for Uncut & OPP. The shelf-life of 2nd grade mushrooms was about 22 days for Cut & Tray, 17 ays for Cut & OPP, and 14 days for Uncut & OPP. Factors affecting fresh mushroom quality included browning of cap and stalk, and mushroom decay index. Browning of the lower part of the stalk, with related color change as noted in a* and b* values were the main factors indicating quality deterioration of king oyster mushrooms.

A Study on the Effect of Spirulina-containing Cosmetics Using Micro-Needle (미세다륜침을 이용한 스피루리나 함유 화장품의 효과 연구)

  • Jung, SuJin;Lee, Hyunjin;Li, ShunHua
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.6
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    • pp.269-276
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    • 2017
  • In this study, a test was conducted to examine the effects of micro-needle therapy (MTS) and cosmetics containing spirulina on improving the skin tone, skin hydration, hyper-pigmentation, and fine lines of middle-aged women in their 40s and 50s and to propose an effective skin care method. The subjects were divided into a control group of eight people who used serum and cream mask that does not contain MTS and spirulina, and an experimental group who used serum and cream containing MTS and spirulina. The test was conducted with six 30-minute sessions for one week. The change in skin was measured over three times in the week of the test, after six weeks of the test, and two weeks after the test had finished. The result showed that after six weeks of the test, in the experimental group, the skin tone (color brightness, saturation brightness, skin brightness) and moisture showed a significant increase (p<0.05), while the number of small hyper-pigmentation and large hyper-pigmentation, as well as the length of crow's feet, melanin, erythema showed a significant reduction (p<0.05). No significant differences in fine lines under the eyes and length of crow's feet were observed. Two weeks after the test was finished, the experimental group showed a significant decrease in the length of crow's feet and melanin (p<0.05), and a significant increase in skin brightness and moisture (p<0.05). Even after the test was completed, the skin brightness, fine lines, melanin, and moisture improved continuously. In conclusion, skin care using MTS and cosmetics containing spirulina was more effective on improving the skin tone, hydration, hyper-pigmentation, and fine lines, than skin care without using them.

Effects of Packaging Materials and Storage Temperature on the Shelf-life of Korean Grinded Yam (포장재료와 보관온도에 따른 갈은 마의 저장수명 및 품질변화에 관한 연구)

  • Ko, Eui Suk;Won, Jong Hwa;Jin, Hyung Seok;Song, Kihyeon;Jeon, Kyu Bae;Kim, Jai Neung
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.20 no.3
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    • pp.103-111
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    • 2014
  • This study analyzed effects of packaging materials and storage temperature on the shelf-life of Korean grinded yam. For experimental measurement, experimental groups were used $PET(12{\mu}m)/Al(9{\mu}m)/LLDPE(60{\mu}m)$ as opaque barrier packaging material and $PET(12{\mu}m)/PE(20{\mu}m)/VM-PET(12{\mu}m)/PE(25{\mu}m$) and barrier $NY(15{\mu}m)/LLDPE(60{\mu}m$) as transparent barrier packaging material. Control group were used normal $NY(15{\mu}m)/LLDPE(65{\mu}m)$. The grinded yam stored at refrigeration temperature ($5^{\circ}C$) and room temperature ($20^{\circ}C$C) for 8 days. Changes in color, pH, viscosity, microorganisms and sensory characteristics were measured. In the grinded yam L and ${\Delta}E$ value were not changed rapidly at refrigeration temperature. However L and ${\Delta}E$ were rapidly changed in all of packaging materials at room temperature. In two temperature condition, pH and viscosity were decreased during storage but pH was increased at $5^{\circ}C$ after 4 days. PET/Al/LLDPE and PET/PE/VM-PET/PE inhibited the growth of microorganisms effectively. PET/AL/LLDPE which is a high barrier packaging material showed the best in two temperature condition.

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Study on Hospital Environmental Causes Affected the Mother′s Comfort After Her Child Birth (산욕부 안위에 영향을 미치는 병원환경 요인에 관한 연구)

  • 변수자
    • Journal of Korean Academy of Nursing
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    • v.8 no.1
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    • pp.1-15
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    • 1978
  • The Purpose of this study is to examine closely the causes influenced upon the comfort and recovery of the woman delivered of a child in the hospital at the same time to understand environmental status of hospitals in order to promote mother's health recovery, and to improve hospital environment by emphasizing the meaning of environment and health before the medical staff and hospital administrative authority. In the method of servery of the research, 165 post paestum patients have been randomly selected who were accommodated and delivered their babies at OB(obstetric ) & GY (Gynecologic) unit the 7 general hospitals for the period of 6 December 1976 through 17 December 1976. As for the survey, it has been used of Questionnaire where we have 65 items in the respect of personal environment in the hospital such as trusting nurse, ability, reliability, kindness and etiquette of nurse and tile character of nurse the relationship with patients the other respect of physical environment included 9f temperature, moisture. air-ventilation lightening noise, cleanness. facilities, and the third realm being of mother's hearth ground to have the following conclusion 1. The feature of the collected personnel they are from OB or GY sects of from OB unit of the other 5 hospitals except the two general hospitals of the college or school Otherwise the rate of the patients to nurses would be 9 : 1. As for the nurses'ground it would be appeared of 20-25 years of age as the 76%. either 3 year course or 4 year course in the education would be each 50% and less than 2 year experience case would record as of 60 %. In the respect of hospital physical environmental status, there we have two hospitals without any thermometers, on the other han4 nowhere there's hygrometer, otherwise, the lightening is normal or over than normal As for the structure of noise protection the corridors're, generally speaking worse than rooms, nerver hueless, there's no ventilating system in the hospitals. The rooms'repainted in white and yellow, light green white, or green color. The patient's clothing were in green pink blue, light green or in white co for. There're not anything special in both decoration and equipments. Most of them used tall beds except in one hospital 2. To the extent of perception of patient's hatch 9round and hospital environment it is presented that they perceived nurse's ability in highest in total human variable, though perceived kindness or etiquette in the lowest otherwise, comparatively high in total average. 3. In the respect of physical environment it is highest perceived of lightening terms, otherwise, lowest perceived of air ventilation and total average became lowest than the one of the original record 4. To ages, in the respect of hatch ground rather old aged mother than the younger one has perceived that nurse would be trusting, in good service character, able, at the same time, liable, Otherwise, in physical environment regardless of age, they perceived lightening in high and remarkably lower in ventilation As a result of the examination of the difference in hospital environment to each age it is appeared of statistical difference at 5% level of ability in the personal environment otherwise little difference as for physical environment 5. In the respect of perceiving level to educational standard it is highly perceived of personal environment for higher ranking group rather than lower group in the educational standard. In case of physical environment it is highly perceived for lower level group rather than higher level group in educational background. The variables which have statistical significance at 5% level are from trusting kindness, etiquette and total kindness, etiquette and total all significance at 5% level are from trusting, kindness, etiquette and total human environment variable in personal environment, otherwise, there's little difference in the physical environment. 6. The perceiving level due to times of admission and accommodation at the hospital would be cleared out as gradual higher perception both physical and personal environment in the hospital. At 5% significant level of the ventilation condition in physical environmental variable it is presented of meaningful difference otherwise, there we have little difference both in Personal variable and other one. 7. In accordance with living standard, the perception degree of personal environment in tee hospital would be inclined to increase to higher living standard on the other hand, in case of the physical environment, the perception level world increase to lower living standard At 5 % level, the trustuariable and total scores in the personal eicuironmectal variable there appeared a meaningful/ significant difference otherwise, there presented little difference both in physical environmental and other variable to the living standard 8. Pertaining to family unit, the mother of an independent family unit perceived highly in all respect of the personal and the physical environment in the hospital rather than the woman of succeeding family unit. At 5 % level there appeared a difference in the respect of kindness and etiquette both in personal environment variable, on the other hand, there hardly marked a difference between other variable and physical environmental one. 9. The degree of perception to comforting level has little connection with a statistical difference the age, educational level hospital admitting times, living standard or family unit. 10. The most effective variable to mother's comforting level will be nurse's ability, reliability, trusting manner, and total physical environment variable in order.

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Application of the CRISPR/Cas System for Point-of-care Diagnosis of Cattle Disease (현장에서 가축질병을 진단하기 위한 CRISPR/Cas 시스템의 활용)

  • Lee, Wonhee;Lee, Yoonseok
    • Journal of Life Science
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    • v.30 no.3
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    • pp.313-319
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    • 2020
  • Recently, cattle epidemic diseases are caused by a pathogen such as a virus or bacterium. Such diseases can spread through various pathways, such as feed intake, respiration, and contact between livestock. Diagnosis based on the ELISA (Enzyme-linked immunosorbent assay) and PCR (Polymerase chain reaction) methods has limitations because these traditional diagnostic methods are time consuming assays that require multiple steps and dedicated equipment. In this review, we propose the use of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) Cas system based on DNA and RNA levels for early point-of-care diagnosis in cattle. In the CRISPR/Cas system, Cas effectors are classified into two classes and six subtypes. The Cas effectors included in class 2 are typically Cas9 in type II, Cas12 in type V (Cas12a and Cas12b) and Cas13 in type VI (Cas13a and Cas13b). The CRISPR/Cas system uses reporter molecules that are attached to the ssDNA strands. When the Cas enzyme cuts the ssDNA, these reporters either fluoresce or change color, indicating the presence of a specific disease marker. There are several steps in the development of a CRISPR/Cas system. The first is to select the Cas enzyme depending on DNA or RNA from pathogens (viruses or bacteria). Based on that, the next step is to integrate the optimal amplification, transducing method, and signal reporter. The CRISPR/Cas system is a powerful diagnostic tool using a gene-editing method, which is faster, better, and cheaper than traditional methods. This system could be used for early diagnosis of epidemic cattle diseases and help to control their spread.

Detection of Helicobacter pylori by Pre-embedding Immunoelectron Microscopy - Comparison with Immunoblotting Method (Pre-embedding Immunoelectron Microscopy를 이용한 Helicobacter pylori의 발견 - Immunoblotting법과 비교 -)

  • Jung, Hae-Seoung;Kim, Eun-Jung;Kim, Eun-A;Park, Ji-Hoe;Jun, Jin-Su;Seo, Ji-Hyoun;Lim, Jae-Young;Choi, Myoung-Bum;Woo, Hyang-Ok;Youn, Hee-Shang;Ko, Gyung-Hyuck;Baik, Seung-Chul;Lee, Woo-Kon;Cho, Myung-Je;Rhee, Kwang-Ho
    • Clinical and Experimental Pediatrics
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    • v.45 no.7
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    • pp.862-874
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    • 2002
  • Purpose : We tried to evaluate whether the detection rate of Helicobacter pylori in gastric biopsy specimens could be improved by using pre-embedding immunoelectron microscopy. Methods : A total of 119 children who complained of upper gastrointestinal symptoms were endoscoped at the Gyeongsang National University Hospital from July, 1996 to July, 1999. Five biopsy specimens(three for urease test, one for hematoxylin-eosin(H & E) staining, and one for preembedding immunoelectron microscopy) were obtained from each antrum and body. Immunoblotting analysis were also performed. Results : Among the 119 patients, H. pylori were found in 116 patients(97.5%) by the immunoelectron microscopy. Among three patients who were found H. pylori negative in immunoelectron microscopy, two patients showed H. pylori in H & E stained slides and one patient was urease test positive(color change within six hours). Urease tests were positive in 107 patients(89.9 %). The positive rate of immunoblotting tests was 81.5%. However, only 13 patients(10.9%) showed H. pylori on the H & E stained antrum or body tissue. Conclusion : In this study, we found H. pylori histopathologically in most of the pediatric patients who complained of upper gastrointestinal symptoms. This study showed that pre-embedding immunoelectron microscopic examinations can be used as a gold standard in the diagnosis of childhood H. pylori infection. However, this method also has limited capacity to detect widely scattered H. pylori compared to the other histopathologic diagnostic methods.

Effects of pH and Redox Conditon on Silica Sorption in Submerged soils (담수조건(湛水條件)에서 토양산도(土壤酸度)와 산화환원(酸化還元) 전위(電位)가 토양(土壤)의 규산흡착(珪酸吸着)에 미치는 영향(影響))

  • Lee, Sang-Eun;Neue, Heins Ulitz
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.2
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    • pp.111-126
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    • 1992
  • Silica sorption isotherm belonged to the C-type with weak L-type characteristics according to the classification system of adsorption isotherm. Silica sorption isothem fitted well to the Freundlich and Tempkin equation but not to the Langmuir equation. The color interference probably due to $Fe^{2+}$ during spectrometric silca determination by Molybdenum-blue method affected the sorption isotherm in reduced soils or low pH. Four parameters such as the intercept of Freundlich equation, the slope of Tempkin equation, the "Silica reactivity", and the "C-type slope", where the last two parameters were termed in the current study, were examined to assess treatment effects on silica sorption. Among them the "C-type slope" was found out to be the best parameter. The C-type isotherms showed the same high correlation coefficient as Freundlich and Tempkin equation when regressed to the sorption isothem. Plotting the C-type slope on a logarithmic scale vs. the pH showed high linearity. Using the "C-type slope" as a perameter, the pH and soil type affected the silica sorption while the effect of redox condtion was not significant. All Fe and Al extracted by the various reagents, and OM were highly correlated to silica sorption. Among them $Fe_d$ was identified as the highest influencing soil property. Since there is no equivalent reliable method to discriminate the forms of the soil Al-oxides their likely importance remains unclear.

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3D Facial Animation with Head Motion Estimation and Facial Expression Cloning (얼굴 모션 추정과 표정 복제에 의한 3차원 얼굴 애니메이션)

  • Kwon, Oh-Ryun;Chun, Jun-Chul
    • The KIPS Transactions:PartB
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    • v.14B no.4
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    • pp.311-320
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    • 2007
  • This paper presents vision-based 3D facial expression animation technique and system which provide the robust 3D head pose estimation and real-time facial expression control. Many researches of 3D face animation have been done for the facial expression control itself rather than focusing on 3D head motion tracking. However, the head motion tracking is one of critical issues to be solved for developing realistic facial animation. In this research, we developed an integrated animation system that includes 3D head motion tracking and facial expression control at the same time. The proposed system consists of three major phases: face detection, 3D head motion tracking, and facial expression control. For face detection, with the non-parametric HT skin color model and template matching, we can detect the facial region efficiently from video frame. For 3D head motion tracking, we exploit the cylindrical head model that is projected to the initial head motion template. Given an initial reference template of the face image and the corresponding head motion, the cylindrical head model is created and the foil head motion is traced based on the optical flow method. For the facial expression cloning we utilize the feature-based method, The major facial feature points are detected by the geometry of information of the face with template matching and traced by optical flow. Since the locations of varying feature points are composed of head motion and facial expression information, the animation parameters which describe the variation of the facial features are acquired from geometrically transformed frontal head pose image. Finally, the facial expression cloning is done by two fitting process. The control points of the 3D model are varied applying the animation parameters to the face model, and the non-feature points around the control points are changed by use of Radial Basis Function(RBF). From the experiment, we can prove that the developed vision-based animation system can create realistic facial animation with robust head pose estimation and facial variation from input video image.

Isolation and Morphological Identification of Fresh Water Green Algae from Organic Farming Habitats in Korea (유기농업 생태계로부터 담수 녹조류 분리 및 형태적 동정)

  • Kim, Min-Jeong;Shim, Chang-Ki;Kim, Yong-Ki;Hong, Sung-Jun;Park, Jong-Ho;Han, Eun-Jung;Jee, Hyeong-Jin;Yun, Jong-Chul;Kim, Suk-Chul
    • Korean Journal of Organic Agriculture
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    • v.22 no.4
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    • pp.743-760
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    • 2014
  • This study aimed to isolate and identify freshwater algae from the organic agricultural ecosystems and investigate its biological characteristics to study the possibility of utilizing a biomass freshwater algae in organic farming. In the survey area, average water temperature was $12.4{\sim}28.2^{\circ}C$ and the pH ranges were from 6.1 to 8.5. The solid culture method is more suitable than liquid culture method for isolation of freshwater algae with lower contamination level and higher isolation frequency. A total of 115 strains were isolated from six freshwater algae habitats in nine regions in Korea. BGMM (BG11 Modified Medium) amended with NaNO3 and $KNO_3$ as a nitrogen, and $Na_2CO_3$ as carbon source was designed to isolate and culture freshwater algae. Absorbance of freshwater algae culture has increased dramatically to four days and decreased after eight days after inoculation. CHK008 of the seven isolates showed the highest absorbance in seven days after culturing in BGMM. The optimal pH of BGMM for culturing freshwater algae was pH 6-7. As light intensity increased, growth of freshwater algae increased. Among the five kinds of carbon sources, glucose and galactose promoted good growth of freshwater algae in BGMM. The colony color of purified 16 green algae isolates showed a separation of green, dark and light green, and of them, eleven algae strains showed a strong fluorescent light under fluorescence microscopy. Cell size of the green algae showed a wide range of variation depending on the species. General morphology of the green algae strains was spherical. Chlamydomonas sp. was elliptical, and Chlorella sorokiniana was ellipsoidal and cylindrical. All strains of the green algae except for Chlamydomonas sp. did not have flagella. One isolate of Chlamydomonas sp. and five isolates of C. sorokiniana secreted mucus. Sixteen isolates of 16 green algae were identified as two family and six species, Chlorella vulgalis, C. sorokiniana, C. pyrenoidosa, C. kessleri, C. emersonii, and Chlamydomonas sp. based on their morphological characteristics.