{"medium":{"id":"78c","name":"CHOPPED MEAT MEDIUM FOR FRETIBACTERIUM SP.","complex_medium":"yes","min_pH":7,"max_pH":7,"source":"DSMZ","link":"https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium78c.pdf"},"solutions":[{"id":133,"name":"Main sol. 78c","volume":853,"recipe":[{"recipe No.":1,"compound":"Ground beef","compound_id":128,"attribute":"fat free","amount":500,"unit":"g","g_l":586.166,"optional":"no"},{"recipe No.":2,"compound":"NaOH","compound_id":256,"attribute":"1 N","amount":25,"unit":"ml","optional":"no"},{"recipe No.":3,"compound":"Casein peptone","compound_id":21,"attribute":"BD BACTO","amount":30,"unit":"g","g_l":35.17,"optional":"no"},{"recipe No.":4,"compound":"Yeast extract","compound_id":16,"amount":5,"unit":"g","g_l":5.86166,"optional":"no"},{"recipe No.":5,"compound":"K2HPO4","compound_id":10,"amount":5,"unit":"g","g_l":5.86166,"mmol_l":33.6537,"optional":"no"},{"recipe No.":6,"compound":"Sodium resazurin","compound_id":1852,"attribute":"0.1% w/v","amount":0.5,"unit":"ml","g_l":0.000586166,"mmol_l":0.00233374,"optional":"no"},{"recipe No.":7,"compound":"L-Cysteine HCl x H2O","compound_id":770,"amount":0.5,"unit":"g","g_l":0.586167,"mmol_l":3.33739,"optional":"no"},{"recipe No.":8,"compound":"Na-fumarate","compound_id":143,"amount":6,"unit":"g","g_l":7.034,"mmol_l":43.9526,"optional":"no"},{"recipe No.":9,"compound":"Na-formate","compound_id":144,"amount":6,"unit":"g","g_l":7.034,"mmol_l":103.43,"optional":"no"},{"recipe No.":10,"solution":"Clarified rumen fluid","solution_id":2618,"condition":"clarified","amount":50,"unit":"ml","optional":"no"},{"recipe No.":11,"compound":"Horse serum","compound_id":146,"condition":"","attribute":"inactivated, INVITROGEN","amount":50,"unit":"ml","optional":"no"},{"recipe No.":12,"solution":"Fatty acid mixture","solution_id":209,"amount":20,"unit":"ml","optional":"no"},{"recipe No.":13,"solution":"Wolin's vitamin solution (10x)","solution_id":5980,"amount":1,"unit":"ml","optional":"no"},{"recipe No.":14,"solution":"Haemin solution","solution_id":124,"attribute":"0.05% w/v","amount":5,"unit":"ml","optional":"no"},{"recipe No.":15,"solution":"Vitamin K1 solution","solution_id":125,"attribute":"0.05% w/v","amount":2,"unit":"ml","optional":"no"},{"recipe No.":16,"compound":"DL-Dithiothreitol","compound_id":151,"attribute":"DTT","amount":0.3,"unit":"g","g_l":0.3517,"mmol_l":2.27933,"optional":"no"},{"recipe No.":17,"compound":"Distilled water","compound_id":4,"amount":700,"unit":"ml","optional":"no"}],"steps":[{"step":"Use lean beef or horse meat. Remove fat and connective tissue before grinding. Mix meat, water and NaOH, then boil for 15 min with stirring. Cool to room temperature, skim fat off surface, and filter, retaining both meat particles and filtrate. To the filtrate add Casitone, yeast extract, hydrogen phosphate and resazurin, then add water to a final volume of 700 ml. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then add cysteine and adjust pH to 7.0. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes containing meat particles (use ca. 7 ml liquid medium for 1 - 2 g meat particles), then autoclave at 121\u00b0C for 20 min. After autoclaving add fumarate, formate, rumen fluid, horse serum, fatty acids, vitamins, haemin, vitamin K1 and DTT rom sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and sterilized by filtration."}],"equipment":{"1296":"Autoclave","2098":"Hungate tubes","3118":"Filter","3961":"Gassing station"},"main solution":"yes"},{"id":2618,"name":"Clarified rumen fluid","volume":1000,"steps":[{"step":"Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121\u00b0C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20\u00b0C."}],"Sol. No.":6,"main solution":"no"},{"id":209,"name":"Fatty acid mixture","volume":1000,"recipe":[{"recipe No.":1,"compound":"Isobutyric acid","compound_id":222,"amount":23,"unit":"ml","optional":"no"},{"recipe No.":2,"compound":"DL-2-Methylbutyric acid","compound_id":223,"amount":27,"unit":"ml","optional":"no"},{"recipe No.":3,"compound":"Valeric acid","compound_id":224,"amount":27,"unit":"ml","optional":"no"},{"recipe No.":4,"compound":"Isovaleric acid","compound_id":225,"amount":27,"unit":"ml","optional":"no"},{"recipe No.":5,"compound":"Distilled water","compound_id":4,"amount":896,"unit":"ml","optional":"no"}],"steps":[{"step":"Adjust pH to 7.5 with concentrated NaOH."}],"Sol. No.":7,"main solution":"no"},{"id":124,"name":"Haemin solution","volume":100,"recipe":[{"recipe No.":1,"compound":"Haemin","compound_id":149,"amount":50,"unit":"mg","g_l":0.5,"mmol_l":0.766989,"optional":"no"},{"recipe No.":2,"compound":"NaOH","compound_id":256,"attribute":"1 N","amount":1,"unit":"ml","optional":"no"},{"recipe No.":3,"compound":"Distilled water","compound_id":4,"amount":100,"unit":"ml","optional":"no"}],"steps":[{"step":"Dissolve 50 mg haemin in 1 ml 1 N NaOH; make up to 100 ml with distilled water and filter sterilize. Store refrigerated."}],"equipment":{"1875":"Filter"},"Sol. No.":9,"main solution":"no"},{"id":125,"name":"Vitamin K1 solution","volume":20,"recipe":[{"recipe No.":1,"compound":"Vitamin K1","compound_id":150,"amount":0.1,"unit":"ml","optional":"no"},{"recipe No.":2,"compound":"Ethanol","compound_id":201,"attribute":"95 %","amount":20,"unit":"ml","g_l":950,"mmol_l":20620.8,"optional":"no"}],"steps":[{"step":"Dissolve 0.1 ml of vitamin K1 in 20 ml 95% ethanol and filter sterilize. Store refrigerated in a brown bottle."}],"equipment":{"277":"Filter"},"Sol. No.":20,"main solution":"no"},{"id":5980,"name":"Wolin's vitamin solution (10x)","volume":1000,"recipe":[{"recipe No.":1,"compound":"Biotin","compound_id":46,"amount":20,"unit":"mg","g_l":0.02,"mmol_l":0.0818632,"optional":"no"},{"recipe No.":2,"compound":"Folic acid","compound_id":138,"amount":20,"unit":"mg","g_l":0.02,"mmol_l":0.0453104,"optional":"no"},{"recipe No.":3,"compound":"Pyridoxine hydrochloride","compound_id":519,"amount":100,"unit":"mg","g_l":0.1,"mmol_l":0.486287,"optional":"no"},{"recipe No.":4,"compound":"Thiamine HCl","compound_id":1207,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.148236,"optional":"no"},{"recipe No.":5,"compound":"Riboflavin","compound_id":136,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.132837,"optional":"no"},{"recipe No.":6,"compound":"Nicotinic acid","compound_id":139,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.406141,"optional":"no"},{"recipe No.":7,"compound":"Calcium D-(+)-pantothenate","compound_id":1512,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.209846,"optional":"no"},{"recipe No.":8,"compound":"Vitamin B12","compound_id":18,"amount":1,"unit":"mg","g_l":0.001,"mmol_l":0.00073779,"optional":"no"},{"recipe No.":9,"compound":"p-Aminobenzoic acid","compound_id":47,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.364591,"optional":"no"},{"recipe No.":10,"compound":"(DL)-alpha-Lipoic acid","compound_id":252,"amount":50,"unit":"mg","g_l":0.05,"mmol_l":0.24233,"optional":"no"},{"recipe No.":11,"compound":"Distilled water","compound_id":4,"amount":1000,"unit":"ml","optional":"no"}],"main solution":"no"}]}