We investigated the optimal culture conditions for Cryptosporidium muris in a human stomach adenocarcinoma (AGS) cell line by determining the effects of medium pH and of selected supplements on the development of C. muris. The optimum pH of the culture medium required for the development of C. muris was determined to be 6.6. The number of parasites significantly increased during cultivation for 72 hr (p < 0.05) at this level. On the other hand, numbers decreased linearly after 24 hr of incubation at pH 7.5. When cultured in different concentrations of serum, C. muris in media containing 5% FBS induced 4-7 times more parasites than in 1% or 10% serum. Of the six medium supplements examined, only 1 mM pyruvate enhanced the number of C. muris in vitro. Transmission electron microscopic observation showed the developmental stages of C. muris in the cytoplasm of the cells, not in an extracytoplasmic location. The growth of C. muris in AGS cells provides a means of investigating its biological characteristics and of testing its response to therapeutic agents. However, a more optimized culture system is needed for the recovery of oocysts on a large scale in vitro.
Mites in the genus Chorioptes cause a mild form of skin disease in both domestic and wild ruminants. In July 2006, dermatitis characterized by alopecia, marked lichenification, accumulation of crust, and fissuring was recognized in 14 out of 200 Holstein dairy cattle raised in the cattle farm of the National Institute of Animal Science in Cheonan, Republic of Korea. Skin lesions were distributed mainly over the tail base, and sacral and perineal regions. Microscopic examinations of skin scraping samples from severely affected areas revealed numerous mites of all developmental stages. Morphologically, pedicels of the mites were short and unjointed. The tarsal suckers occurred on the pedicels of all the legs in the male worm and on the first, second, and fourth pair of legs in the adult female worm. A single long seta at the tarsus of legs III and the length of legs II being about twice as long as legs IV in adult male mites were observed. Arising anterior to the inner-most spatulate seta was a short seta with an average of $26.4{\pm}5.8{\mu}m$ in length. Also, the length of setae #4 on the opisthosomal lobes was relatively short. Based on these observations, the mites were identified as Choriptes texanus. Although the chorioptic mange may not influence the mortality rate in the affected farm, reports indicate that a decline in milk production can be observed. This is the first report of chorioptic infestation in a cattle farm from Korea.
Bang, Sookie S.;Anderson, Cynthia M.;Bergmann, David J.;Sieverding, Heidi L.;Flanegan, Amy L.;Braaten, Amanda S.;Masteller, Amanda R.
Proceedings of the Microbiological Society of Korea Conference
/
2008.05a
/
pp.118-118
/
2008
The phylogenetic diversity of microbial communities in calcareous mats from Spearfish Creek, a freshwater stream located in the Black Hills of South Dakota, was examined using PCR-based 16S rDNA sequence analysis. In this study, two types of calcareous mats were compared: mature mats formed on the natural substrate of rock surfaces and developing mats on an artificial substrate of glass slides. Among 63 unique isolates from a clone library of 16S rRNA genes from mature mat samples, there were 8 phyla of Bacteria represented. The predominant phylum was Proteobacteria (48%), with the $\beta$ subclass being the largest group. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes from slide samples collected at intervals for four months showed considerable diversity of the microbial community from the earliest stages of community development. Amplicons isolated from DGGE gels and sequenced indicated that community succession has occurred without increasing microbial diversity. However, light microscopic analysis revealed a significant increase in microbial cell density throughout the community development. Scanning electron microscopy of mat samples provides evidence that diatoms are also important members of calcareous mat communities.
Premitotic, mitotic and postmitotic nuclei in the dikaryotic somatic hyphae of Pleurotus ostreatus, the oyster mushroom fungus were ultrastucturally examined using chemical fixation and freeze-substitution process, and the behaviors of astral microtubules associated with these nuclei were closely analyzed. Electron microscopic examinations revealed that astral microtubules are significantly abundant when the nuclei are in the stage of migration and at the stage of migration, the separation of spindle pole body occurs. Such an abundancy of astral microtubules in premitotic migrating nuclei is well contrasted with mitotic and postmitotic nuclei with much fewer astral microtubules and it should be noted that neither of these latter classes of nuclei exhibits the separation of the spindle pole body. It is remarkable that the postmitotic nuclei that are believed to migrate actively are associated with the astral microtubules that are less in numbers and length. During all the stages of nuclear division, astral microtubules are invariably radiating from the spindle pole bodies and nucleolus remains within the nuclear envelope of dividing nuclei throughout the division. The functions of astral microtubules developed during the nuclear division as well as the nuclear migration and separation of the spindle pole body were closely examined.
Roh, Young-Bok;Chung, Kyung-A;Chung, Ji-Sook;Kim, Jung-Sam;Kim, Il
The Korean Journal of Zoology
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v.38
no.3
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pp.388-394
/
1995
We investigated ultrastuctural change of small intestinal mucosal epithelial cell, columnar cell and mucous cell, of hibernating ground squirrel during activating and hibernating stages. In active columnar cells, many mitochondria and rough endoplasmic reticulum were observed. In hibernating columnar cells, more free nhosome than rough endoplasmic reticulum were observed. In active mucous cells, large and many mucosal granules, mitochondria and rough endoplasmic reticulum were observed. Mucosal granules have been secreted excellently. In hibernating mucous cells, small and little mucosal granules and many free ribosome were observed.
Park, Yong-Eui;Yang, Deok-Chun;Park, Kwang-Tae;Soh, Woong-Young;Hiroshi Sano
Proceedings of the Botanical Society of Korea Conference
/
1999.07a
/
pp.85-89
/
1999
Somatic embryogendesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology. This paper describes the direct somatic embryogenesis from zygotic embryos of Panax ginseng is reversely related to normal axis growth of zygotic embryos by the experiment of various chemical treatments. Under the normal growth condition, the apical tips of embryo axis produced an agar-diffusible substance, which suppressed somatic embryo development from cotyledons. Although the cells of zygotic embryos were released from the restraint of embryo axis, various factors were still involved for somatic embryo development. Electron microscopic observation revealed that the ultrastructure of cells of cotyledon epidermis markedly changed before initiation of embryonic cell division, probably indicating reprogramming events into the cells embryogenically determined state. Polar accumulation of endogenous auxin or cell-cell isolation by plasmolysis pre-treatment is the strong inducer for the somatic embryo development. The cells for the process of somatic embryogenesis might be determined by the physiological conditions fo explants and medium compositions. Direct somatic embryos from cotyledons fo ginseng were originated eithrer from single or multiple cells. The different cellular origin of somatic embryos was originated either from single or multiple cell. The different cellular origin of somatic embryos was depended on various developmental stages of cotyledons. Immature meristematic cotyledons produced multiple cell-derived somatic embryos, which developed into multiple embryos. While fully mature cotyledons produced single cell-derived single embryos with independent state. Plasmolysis pretreatment of cotyledons strongly enhanced single cell-derived somatic embryogenesis. Single embryos were converted into normal plantlets with shoot and roots, while multiple embryos were converted into only multiple shoots. GA3 or a chilling treatment was prerequisite for germination and plant conversion. Low concentration of ammonium ion in medium was necessary for balanced growth of root and shoot of plantlets. Therefore, using above procedures, successful plant regeneration of ginseng was accomplished through direct single embryogenesis, which makes it possible to produce genetically transformed ginseng efficently.
The purpose of this study was to compare effects of the bioceramics on healing processes of the alveolar bone defects in dogs. Five adult dogs aged 1 to 2 years were used in this study. Experimental alveolar bone defects were created surgically with a #1/2 round bur at the furcation area of the buccal surface of the mandibular 3rd, 4th premolars and 1st molar. Fifteen experimental alveolar bone defects were devided into three groups according to the type of graft materials. The groups were as follows : 1) flap operation with dense hydroxyapatite( DHA group ) 2) flap operation with porous hydroxyapatite( PHA group ) 3) flap operation with natural coral ( NC group ) At 1, 2, 4, 6, and 12 weeks, dogs were serially sacrificed and specimens were prepared with Hematoxylin-Eosin stain and Mallory stain for light microscopic evaluation. The results of this study were as follows : 1. In every group, inflammatory cell infiltrations were seen at 1st weeks due to surgical trauma, however inflammatory response owing to graft materials were not seen. 2. In every group, the appearance of connective tissue around graft materials was loosely formed at the initial stages, however the connective tissue was densely formed at 2 weeks. 3. The presence of osteocytes were observed at 2 weeks in the natural coral group, however the osteocytes were appeared at 6weeks in the dense hydroxyapatite group. 4. A new bone was formed from the base and walls of the defect and gradually expanded toward the graft materials. 5. A resorption of the natural coral occurred irregularly at the periphery of the material, therefore the size and shape of the natural coral were reduced at 6 weeks. 6. At 12 weeks, the porous hydroxyapatite and natural coral were surrounded by newly formed bone most completely, however dense hydroxyapatite was surrounded by newly formed bone in part.
The purpose of this study was to investigate effects of the natural coral(NC) and the hydroxyapatite/calcium sulfate hemihydrate(HA/CS) on an early stages of wound healing in the rat periodontal fenestration defects. In this experiment, twelve male rats(Mean : 520g in BW) aged 8 to 9 months were used. Experimental periodontal fenestration defects were surgically created with tapered fissure bur at the buccal surface of the left mandibular 1st, 2nd molars. The buccal aspects of molar roots were carefully denuded of their periodontal ligament through a bony window created in the left mandibles of rats under general anesthesia. Each experimental periodontal fenestration defect was grafted with natural coral and HA/CS, randomly. An area without bone graft was assigned for negative control group. At 10,35 days, rats were serially sacrificed via intracardiac perfusion with 2.5% glutaraldehyde and specimens were processed with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows : 1. The defect areas were filled with dense connective tissues at 10 days in control group. But in the test(NC, HA/CS)groups, the connective tissues around graft materials were formed more loosely and the response of inflammation by graft materials itself was not found. 2. The defect areas were filled with new osteoid tissues and new cementum was not formed on the cut root surface at 35 days in the control group. 3. New osteoid tissue formation was more prominent at 35 days in control than test groups. 4. The NC and HA/CS particles were encapsulated by loose connective tissues at 10 days and by dense connective tissues at 35 days, respectively. 5. In the test groups, resorption of graft particles was not found through the experimental time. From the above results, natural coral and hydroxyapatite/calcium sulfate hemihydrate may be biocompatible and osteoconductive and have a weak adverse reaction to the periodontal tissues.
Bacteria are one of the most important causative agents of the pulpal and periapical diseases. Streptococci are one of the most frequently isolated facultative anarerobic bacteria in the infected root canals. Bacterial cell wall components have a direct effect in the pathogenesis of the pulpal and periapical infections. Hyaluronidase produced by bacteria has been implicated in dissemination of the diseases. The purpose of this study was to evaluate the effect of cell wall extract of streptococci on the L929 cells using inverted microscope and the transmission electron microscopy (TEM). Hyaluronidase production of streptococcal strains were investigated to determine the correlation between the severity of cell damage and the activity of enzymes. Bacterial cell wall extracts of S. sanguis, S. mitis and S. uberis isolated from infected root canals and ATCC type strains of S. mutans (ATCC 10449) and E. faecalis (ATCC 19433) were prepared by sonication and confirmed with SDS-PAGE. Silver stain of SDS-PAGE of sonic extract was efficient at $100{\mu}g$/ml concentration of cell wall protein, while Coomasie blue stain was efficient at $100{\mu}g$/ml concentration. Inverted microscope showed that sonic extract-treated L929 cells were round and detached from the substratum while others lost their fibroblastic shapes. Transmission electron microscopic examination revealed that streptococcal extracts induced death of L929 cells. Sonic extracts of streptococci had variable effect on microstructure of L929 cells. significant chromatin condensation was observed in the nucleus of the cells. Disappearance of cell surface microvilli and nuclear fragments with dense chromatin were observed. The cell nucleus had an irregular shape and numerous large vacuoles were seen in the cytoplasm and some breaks of the cell membrane could be seen. Cell organelles were in various stages of destruction and cristae of mitochondria were disoriented or disappeared. Eighteen strains of streptococci did not produce hyaluronidase.
Pathological interrelations of two soil-borne diseases in cucurbits (watermelon, oriental melon, shintosa and cucumber) caused by Fusarium isolates (FI) and the root-knot nematode (RKN), Meloidogyne incognita were characterized by the fusarium disease severity index (DI), RKN gall index (GI) and eggmass index (EI) in inoculation tests using FI and RKN. Virulence of FI as determined by DI at 4 weeks after inoculation was mostly in the higher order of Fusarium proliferatum F6, F5 and Fusarium oxysporum f. sp. melonis or Fusarium oxysporum f. sp. niveum with no significant differential interactions among the cucurbits and RKN co-infection. Significant increases of DI due to RKN coinfection were noticed in watermelon and oriental melon infected with F. proliferatum isolates, suggesting the DI increase due to RKN coinfection may depend upon the virulence of FI relative to aggressiveness of RKN on the cucurbits. For the coinfection of FI and RKN, GI and EI were mostly reduced logarithmically with the increase of DI, largely more in EI than GI, in all cucurbits except for shintosa. Microscopic examination of the root tissues showed histopathological features characteristic to infection types; formation of fungal hyphae and/or spores and plant defense structures (tyloses and mucilage) in variable degrees and formation of giant cells at variable developmental stages and with variable cytoplasmic depletion or degeneration which were visualized in relations with the values of DI, GI and EI. These findings will be helpful to develop control strategies of the soil-borne disease complex based on their pathological characteristics.
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