Willaertia magna De Jonckheere et al. 拉丁名
(ATCC® 50035™) 統(tǒng)一編號
Strain Designations 別名 Z503
Biosafety Level 生物安全等級 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Isolation 分離基物 Bovine feces,
France Product 提供形式 Format frozen
Storage Conditions 生長條件
Frozen Cultures 冷凍物: -70°C for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures 凍干物: 2-8°C
Cultures: See Protocols section for handling information
Type Strain 模式菌株 yes
Comments 注釋 Isoenzyme electrophoresis Occurrence in natural waters in Mexico Review
Medium 培養(yǎng)基 ATCC® Medium 1034: Modified PYNFH medium (Available from ATCC as ATCC cat. no. 327-X)
Growth Conditions 生長條件
Temperature 培養(yǎng)溫度: 35°C
Culture System: Axenic
Cryopreservation
Harvest and Preservation
1，To achieve the best results set up cultures with several different inocula (e.g. 0.25 mL, 0.5 mL, 1.0 mL). Harvest cell cultures and pool when the lowest inoculum is at or near peak density.
2， the cell concentration exceeds the required level do not centrifuge, but adjust the concentration to between 2 x 106 and 2 x 107cells/mL with fresh medium. If the concentration is too low, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield the desired concentration.
3，While cells are centrifuging prepare a 15% (v/v) solution of sterile DMSO as follows: Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath. Allow the DMSO to solidify. Add the required volume of refrigerated medium. Dissolve the DMSO by inverting the tube several times. *NOTE: If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.
4，Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be between 106 and 107 cells/mL and 7.5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no less than 15 min and no longer than 60 min.
5，Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
6，Place the vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion. At -40°C plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus. Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1°C/min.)
7，The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer.
8，To establish a culture from the frozen state place an ampule in a water bath set at 35°C (2-3 min). Immerse the vial just sufficient to cover the frozen material. Do not agitate the vial.
9，Immediately after thawing, aseptically remove the contents of the ampule and inoculate into 10 mL of fresh ATCC medium 1034 in a T-25 tissue culture flask. Incubate at 35°C.
Name of Depositor 寄存人 JF De Jonckheere
Chain of Custody 來源國家 ATCC <-- JF De Jonckheere <-- M. Pussard
References 參考文獻 Pussard M, Pons R. Etude des pores kystiques de Naegleria (vahlkampfiidae - amoebida). Protistologica 15: 163-175, 1979. Environ. Res. 59: 223-226, 1992.
De Jonckheere DG, et al. Willaertia magna gen. nov., sp. nov. (Vahlkampfiidae), a thermophilic amoeba found in different habitats. Protistologica 20: 5-13, 1984.
John DTOpportunistically pathogenic free-living amebaeIn: John DTParasitic protozoa2nd ed.3San DiegoAcademic Presspp. 143-246, 1993
De Jonckheere JF. Variation of electrophoretic karyotypes among Naegleria spp.. Parasitol. Res. 76: 55-62, 1989. PubMed: 2622896
Rivera F, et al. Pathogenic amoebae in natural thermal waters of three resorts of Hidalgo, Mexico. Environ. Res. 50: 289-295, 1989. PubMed: 2583075
Rivera F, et al. Contaminacion del liquido cefolorraquideo de un infante con sindrome de Arnold-Chiari tipo II, hidrocefalia y mielomeningocele, por Naegleria lovaniensis. Rev. Enferm. Infecc. Pediatr. 2: 91-94, 1989.
