User Manual Systems
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1. Each time the button is pushed the level is increased by one and one more light emitting diode on the LED bar will illuminate If the level is changed this level is set until the system is switched off on or a new level is selected The level can be selected from level one up to level ten indicated by the LED on the indicator panel Table 5 Levels of quality assurance Test Test Test Time of catalyst test Jarleak test Gasinput test Jarleak test Gasinput test Catalyst test 180Sec incl Extended leaktest Jarleak test Gasinput test Catalyst test 75Sec incl Extended leaktest Jarleak test Gasinput test Catalyst test 105Sec incl Extended leaktest Jarleak test Gasinput test Catalyst test 135Sec incl Extended leaktest Jarleak test Gasinput test Catalyst test 165Sec incl Extended leaktest Jarleak test Gasinput test Catalyst test 195Sec incl Extended leaktest Jarleak test Gasinput test Catalyst test 225Sec incl Extended leaktest Jarleak test Gasinput test Catalyst test 255Sec incl Extended leaktest Jarleak test Gasinput test Catalyst test 285Sec incl Extended leaktest We always advise level 2 The level of the quality assurance test is displayed on the indicator panel see Figure 3 Under certain circumstances users might prefer to do a more or a less severe catalyst test In case of new and reactivated catalysts the test may pass quicker than the selected test level thu2. Are you Sure Settings are being saved O Lo D 9 rt 9 v 3 9 o o Mart Microbiology BV Page 20 17 9 2004 Copyright Mart User Manual Run a recipe With the Anoxomat systems there are 2 standard recipes buttons one for the anaerobic atmosphere and the other for a micro aerophilic atmosphere in the jar When one of these buttons is pushed in the operational mode the actions described in the standard recipe are performed A user defined recipe is activated with the DO button from the operational mode Summarising Anaerobic Cycle standard anaerobic recipe Microaerophilic Cycle standard micro aerobic recipe Do default user defined recipe Selection of a user defined recipe Recipe selection is done in the programming mode on menu level Select recipe In the next table an example with the detailed actions to select recipe 17 is shown Table 9 Select recipe 17 a Program mode Until select recipe Enter new menu level EE Until Mod is on display EE Select menu item Mod 0 Until Mod 1 is on display B Select menu item Mod 1 on display Until Mod 17 is on display Select menu item Mod 17 on display e Confirm Esc Operational mode Pushing the Do button in the operational mode will execute recipe 17 Caution Recipe 17 becomes default and remains default until a different recipe is selected Programming a recipe Procedure In
3. panel In contrast to the other activities described in this chapter this is an operational mode action This means that you do not tip the Esc touch switch to enter the programming mode Table 7 Change the intensity of the display Operation increase display intensity Multiple t decrease display intensity The intensity of the display increases decreases step by step with every push on the corresponding button Changing the pumptime jar To check whether a jar is connected to the respective jar tube the Anoxomat will pump on each tube a few seconds thereby sensing the presence or absence of a jar on the tube Normally this pumptime jar is set on 0 8 seconds When using very big or thirdparty jars these 0 8 seconds may not be enough to sense the presence of the jar This may also be the case when the pump gets old The customer can easily check and change this feature Table 8 Change the pumptime jar Programming mode Till menu Select menu Till Program settings Select Program settings Till Pump time jar se Select Pump time jar se Till Mod Select item Mod Till first digit Select first digit Till second digit Select second digit Till third digit Select third digit Till Back Select Back Till Back Select Back Till Program system Select Program system Till Save Select item Save On the display
4. Airtight i Jar1 2or3 ii Indication if the anaerobic jar is airtight or t 1 Jar tube s up to 3 Jars aca O hi a He a Inside diameter 6 mm ae i airtight red not airtight 2 Connection vacuum pump Vacuum A a Inside diameter 8 mm i 1 2 T is righ b Length max 2 m ii Indication K pa e right 3 Connection gas cylinder up to 3 gasses Gas JL AE Ala t red to much or not enoug a Inside diameter 8 mm c y y i Jar1 2or3 b Length max 2 m H EA RES RE i Indication if catalyst activity is sufficient 4 Fuse 160mA slow version 220V ii r de Fuse 315mA slow version 110V a L su sufficient red not sufficient 5 Power supply vacuum pump PUMP i Jart 20r3 6 Power supply 220 110 Volt POWER ii green passed red failure 7 2x fuses inside 2 x 6 3 A 8 On Off switch Installation 1 Connect the Anoxomat to a socket 2 Plug the power cord of the vacuum pump in the back of the Anoxomat 3 Push on the tubes between a the Anoxomat and the vacuum pump Mart Microbiology BV Page 9 17 9 2004 Copyright Mart User Manual Programs G a n n Pa O rosa O Quality assurance Q Anaerobic Cycle O Microaerophilic Cycle 1 Reset a Reset program and errors b Standard settings are kept Quality Assurance a To engage press 1 to10 times before the anaerobic or micro aerophilic program is started Jar LED s light up accordingly b
5. be used 0 no gas 1 anaerobic gas mixture 2 CO for Anoxomat models WS8080 and WS9000 3 Nz only for WS9000 Until which pressure has to be evacuate to setpoint or evacuated or replaced replace to setpoint xxxx digits xxxx in mbar pressure With the parameters named above we have to construct the recipe In total 40 recipes can be programmed by the user The maximum number of action lines for one recipe is 25 Mart Microbiology BV Page 22 17 9 2004 Copyright Mart User Manual Construction of a recipe table Table 11 Example of a recipe table 200 to 1000 Initially the atmospheric air in the jar has to be analysed in order to determine the exact gas composition created by a certain recipe At first the atmosphere in the jar is the same as the atmosphere in the laboratory environment It is assumed that it is composed of 80 Nz and 20 Os The requested low oxygen concentration recipe can be made in one cycle by evacuating the air reduced oxygen tension and then replace it with the anaerobic gas mixture Table 12 Running a low oxygen tension recipe Mod Action Mod Gas Mod Setpoint 1 Low set point descriptions evacuate 0 vacuum leak test replace with No replace with CO 0 end of recipe The setpoints have to be given in mbar of the atmospheric pressure 0 0 3 Intermediate set point 2 High set point 1000 0 Calculation of the setpoints The oxygen concentrati
6. not prereduced contain oxygen which will come in the jar atmosphere during incubation In the literature the following statements about the catalyst are made e the pellets can be inactivated by excessive moisture and H2S therefore they should be reactivated after each incubation by heating the sachet of pellets to 160 C in a drying oven for 1 1 2 to 2 hours e Reactivated catalysts should be stored in a dry area until used Compared to gas generating sachets the Anoxomat has e alower moisture production in the jars e less heat production in the catalyst The catalyst should be renewed regularly because irreversible degradation of catalytic activity takes place due to chemical binding with S ions The end of life period depends on the type of bacterium cultivated or more accurate the H2S production Therefore it cannot be indicated exactly how many incubation cycles can be executed with one sachet Programming panel With the programming panel Figure 4 all variable settings of the Anoxomat system can be changed ut activity Figure 4 Programming panel WS8000 WS8080 and WS9000 With the programming panel a high flexibility of the Anoxomat is achieved Please find hereafter the menu structure used for the Anoxomat Mart Microbiology BV Page 17 17 9 2004 Copyright Mart User Manual Figure 5 Programming tree Menu Program sensor Reset sensor Are you sure Do not do Filter borde
7. 1 MART Microbiology BV User Manual Systems Copyright Mart User Manual Mart Microbiology BV Page 2 17 9 2004 Copyright Mart User Manual Table of Contents TABLE OF CONTEN Ts is sisi i lt s s ses d yeda n dea c s s de e se s de esa ye e e ca des soti Seve a se w s ti s Ve dseo de sese s la 3 USER MANUAL ANOXOMAT i44 452 lt 5345452s 5es saze se e so sae beb eeh ek ke si d oa dewe ek e s d ka 34c d 6 s S ce dede da 3 ses 0 5 INTRODUCTION G yxy54 2212 051325 xad Y vel u te ESAS 5 CONFIGURATION OF THE ANOXOMAT SYSTI lt s scss cssssssesssc sese se sese sesesesesose sense sen re k n U HKH WE E 6 ANOXOMAT SYSTEM BADE docs 6 Ei re 1 gt AnUoxomd SyStem WS8080 ii2 n wa ken n ye nar a RAE 05 nei a RO HEY Ray ENA he a t yan na 6 JANO IN AA O Va KES CEES 0O S ASA EE O VONO SE HAS RE RON W R 7 ANOXOMAT CHARACTERISTICS ia cid 7 Papel CHAT ACIEVISTICS o ok A OA A a e a ado e do e tela ee LEI OS 7 ANOXOMAT TECHNICAL SPECIFICATIONS ococococononononononcnononononononnnnononononononononnnnnnnnnononnnnnnnnnnnnnnnnnnnnnnnononanonananananena 7 Table 2 Electrical speCifiC Gti OM di 7 REDUCING VALV E ras 8 Table 3 Technical requirements of the reducing valve U L L L c cono crono nora nonn nono nnnn cnn naar cann k
8. 4 54 54 55 c4 sei sis es sa d e sa a eke cide eo o i eo d e d ze s e d qe 3 sk U0 e e da e b e Ze s e s s ck dede e2 eke 17 Figure 4 Programming pane WS8000 WS8080 and WS9000 uE ake ek aye axek ek kk e keke 17 Figure 3 PrOgramming lts dsd Hey eg ee kk HU as aa Le nnd oa dkan a e veka eh aWak can e Bi Hek ED MUA k E 18 CHANGING THE NUMBER OF EVACUATION REPLACEMENT CYCLES keke erkeke keke keke keke k k KA KA KA KA KA KA KA KA KA KA KA KA KA 19 Table 6 experiment change the cycles of the anaerobic recipe E L L L LLcL Eek cnn cnn 19 CHANGING THE DISPLAY INTENSITY lt 2o 5 oeae ye y pa EKEN k sir SR Ve A SEN V We we ke pe bee ND ON bU H 9U eu bek Ey V na ewe be e keke ey ee we ley bes v ya 20 Table 7 Change the intensity of the display LL LEE Eek axek nono n cnn crnn Kek kk kk 20 CHANGING THE PUMPTIME TAR cuece vede e sey key wee eq ey k y dde 20 Mart Microbiology BV Page 3 17 9 2004 Copyright Mart User Manual Table 8 Change the pumplime Ji iii 20 RUN A RECIPES td N R rrr dd da rota Ei 21 SELECTION OF A USER DEFINED REGIPE 5551 5355 15 coved scsceveve aly VE Dila a la y l aant 21 Table 9 Select recipe dic hk al l kai a Wak Dav A 21 PROGRAMMING A RECIPE 5 5ii 4si s l k k a3250
9. 5 5l k ke k692 44 ssaa 403 s P l a P l Wa G V E GE PEAR ESS k LAS 0k 5454055550445548 4 21 PROCEDURE 2 a E A RN 21 DEFINITION OF THE GAS COMPOSITION coccccconononononononononononononononononnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnonononononononanonanenenenen 22 RECIPE PARAMETER Siete s 22 Table DO PATGMETECIS tesi tido 22 CONSTRUCTION ORA RECIPE TABLE sdrotoconacodad cidcid dll eee W sae 6u c ba b eyb 6 niba yi 23 Table II Exampleof atecipetable ss iil a y ek eyle n ener x e ere bin bela 0saba Sek k Weye n i Oa d y kn 23 Table 12 Running a low oxygen tension recipe L L L LELE use cono nn neon neon neon nena nc nn nena cria Ku KA 23 CALCUEATION OF THE SETPOINTS vostra ideal dat ara datada 23 PROGRAM DESCRIPTION AND PROGRAMMING oocococonononononononononononononnnnonononnnnnnnnnononnnonnnononnnnnnnnnnnonononononanannnnnanananan 24 Table 13 Programming steps for recipe 17 with 5 0 10 CO and 85 Ng u eee 24 CORRECTION MODE DODO Kop ae aeaa ae asa ae Ee 24 Table 14 Correction mode viccccccccccccssssccccccccsssesceccccccuusseceecccecsuusseccecccessusuceseccceusaussecccessessauseccscessuuaaenseceeeees 24 PROGRAMMING A RECIPE uuee erkeke keke keke keke ke kek keke k ka ka ku ke ku ku ku ku ku ku ka ku ku ku kk ku k kk ka ka k kk k kk keke ke keke Ke Ke KK KK KK KK KK KK KAK A KA KA 25 Table 157 Programming recipe iii huyin
10. Anaerobic yes Yes yes The gas connections of the Anoxomat are non specific i e it does not matter what kind of gasses are connected to the system In the Mart Microbiology B V manual gas supply 1 is always the anaerobic gas mixture gas supply 2 is CO gas and gas supply 3 is N gas 2 Anaerobic gas mixture 80 to 90 N 5 to 10 CO 5 to 10 Ho The Mart Anoxomat quality assurance system consists of Jar leak test Gas input and pressure test Catalyst activity test Extended Jar leak test Anoxomat technical specifications Table 2 Electrical specification Value Value 20 230 V see rear plate 110 120 V see rear plate 50 to 60 Hz 50 to 60 Hz 160 mA slow 315 mA slow Fuse 2 3 6 3 A slow 6 3 A slow Mart Microbiology BV Page 7 17 9 2004 Copyright Mart User Manual Reducing valve Table 3 Technical requirements of the reducing valve 0 4 bar NEN 3268 S 3 8 left turning helix anaerobic gas G 3 8 right turning helix CO Ques 3 8 right turning helix N 2 3kg Gas The standard gas mixture used for anaerobic bacteria cultivation as well as for micro aerophilic bacteria cultivation is made of Table 4 Mixgas composition Ne Nitrogen y 80 to 85 anaerobic basic gas 5 to10 growth stimulation Ha Hidrogen 5 to10 catalysis Installation For details of installation please consult the instruction card hereafter which is also del
11. BV Page 25 17 9 2004 Copyright Mart User Manual Mart Jar system Jar sizes Table 16 Specifications of Mart anaerobic jars AJ9023 AJ9026 AJ9027 AJ9028 240 240 240 240 320 320 320 320 123 140 190 230 154 170 220 260 2 9Lt 3 8Lt 6 9Lt 10 3Lt 1 1 2 3 EEEN Z _ XEM 1 1 2 3 Jar Material Jar and Lid material Transparent PMMA Poly Methyl Meth Acrylic or Cast Acrylic The extremely solid durable construction guarantees a long life The Mart jarlid clamp construction meets the highest quality requirements mechanical as well as microbiological The Mart Jars are never to be cleaned with alcohol and not to be cleaned under temperatures of more than 80 C Please view under Jar cleaner and disinfectant Jar spare parts All jar parts are supplied as spare parts Jar Lid Rubber O ring Catalyst clip s Stainless steel jar clamp with white coating Snap shut coupling for Anoxomat Jar cleaner and disinfectant The Mart Jars are never to be cleaned with alcohol or to be put in the autoclave Mart Microbiology B V advises Halamid Jar cleaner and disinfectant Available in 2 5 500 litres or 5 kg 1 000 litres buckets Based on latent chlorine and oxygen Effective against bacteria fungi and viruses Solution 0 5 Mart Microbiology BV Page 26 17 9 2004 Copyright Mart User Manual Jar maintenance To be certain that the jars are maintained correctly and regularly a small box with nece
12. Following tests are performed c Level 1 bottom LED lights up Jar vacuum leak test gas input test d Level 2 Jar vacuum leak test gas input test catalyst test Standard level 180 sec e Level 3 Jar vacuum leak test gas input test catalyst test 75 sec f Every higher level 30 sec longer catalyst test Anaerobic cycle a Choose the required Quality Assurance level view point 2 b 3 Evacuations and refills to atmospheric pressure c With Catalyst the oxygen level will be 0 Microaerophilic cycle a Choose the required Quality Assurance level view point 2 b 1 Evacuation and refill to atmospheric pressure c Oxygen level 6 Programming facilities ut activity The Anoxomat models WS8000 WS8080 and WS9000 are provided with a programming panel and a display to allow the user to program up to 40 recipes 1 2 Display a Atmospheric pressure b System pressure during operation Buttons a Esc i Switch from operation to programming mode b Ty i step up or down for menu level ii not all levels appear by stepping down c Do i Confirm selection ii Run recipe Trouble shooting 1 No start up a No electricity b Fuses are broken c System error 2 A red LED lights up at Reset a Remaining vacuum in the jar i On digital display closed not at WS80 b Vacuum pump not switched on or connected c Vacuum pump defect d Gas cylinder closed e Jar cap
13. IOLOGY B V product specifications and capabilities However MART MICROBIOLOGY B V cannot guarantee the accuracy of printed materials and disclaims liability for changes errors or omissions Mart Microbiology BV Page 29 17 9 2004
14. Mart User Manual Guarantee LIMITED GUARANTEE ON MART MICROBIOLOGY B V PRODUCTS MART MICROBIOLOGY B V guarantees the original consumer purchaser that this MART MICROBIOLOGY B V product shall be free from defects in materials and workmanship for a period of one 1 year after the original date of purchase At its option MART MICROBIOLOGY B V will repair or replace at no charge any MART MICROBIOLOGY B V product which proves to be defective within such warranty period This limited warranty shall not apply if the MART MICROBIOLOGY B V product has been damaged by unreasonable use accident negligence service or modification by anyone other than a MART MICROBIOLOGY B V Factory Authorized Service Centre or by any other cause related to defective materials or workmanship The original consumer purchaser must return the guarantee card as a condition precedent to warranty coverage and performance To receive warranty service a defective product must be received at a MART MICROBIOLOGY B V Factory Authorized Service Centre no later than one 1 week after the end of the warranty period Purchasers must prepay all delivery costs or shipping charges to return any defective MART MICROBIOLOGY B V product under this warrant policy If you ship any such product we suggest you to pack it securely using the original packing materials if possible and insure it for value as MART MICROBIOLOGY B V assumes no liability for loss or damage
15. a ayibe y anya lay ance fash V n sik b Weney iii 25 MART JARSYSTEM RA eek n k v d ke n de e dU de da k n s kd ke toon e S d deda ue k ke k V d ke Wa e ke d did d s n k 26 JAR SIZES NN 26 Table 16 Specifications of Mart anaerobic j TS L L L L LEE asa c noc kk conc yek nn nr nn ka kK ka ku KK 26 TARMA ERA cd 26 TAR SPAREPAR Tdi 26 JAR CLEANER AND DDISINFECTAN Tot tddi 26 JAR MAINTENANCE oie 27 REFEN CI Sy 44535c 0345532 E reya A aa a V W a la eva yan V dO Ev w WE RA d NEWE W en d 30 u re u W E V E RE w y B b 28 GUARANTEE bes 3533 14 4ye E 00580 k S kab R EKE W E K S WA D RE N ORE RO E O ME Q20 00 0 SE O VV9 K 00095 y s n EK 29 Mart Microbiology BV Page 4 17 9 2004 Copyright Mart User Manual User Manual Anoxomat Introduction The MART Anoxomat system is an automated system which creates an anaerobic micro aerophilic or capnophilic atmosphere in an anaerobic jar just by a push on the button and in about one minute time Evacuating the jar and filling it again with an oxygen free gas mixture according to the method of Macintosh and Fildes create these atmospheres The MART Anoxomat system controls the process and performs a quality assurance test to ensure that all conditions for optimum bacterial growth are realized This manual is meant for us
16. acity more than 8 dm 3 Jar leak Airtight a Jar not airtight b Lid not airtight c Jar coupling not airtight d O ring not sealing off e Stainless steel jar clamp not turned tight 4 A red led lights up at Gas input a Gas pressure to high or to low i Check reducing valve on gas cylinder b Gas cylinder closed i Check reducing valve on gas cylinder c Leak in gas tube or at connections d No gas connected 5 A red LED lights up at Catalyst activity a No Catalyst used with QA level 2 and up b Catalyst used with Microaerophilic cycle c Bad quality of catalyst i reactivate catalyst ii replace catalyst 6 Atmospheric pressure on display lt 800 mbar or gt 1200 mbar a not with WS80 no display b Display reads pressure failure 7 No green led at Passed a Program error occurred b System error occurred User Tips 1 Anoxomat a Always perform Jar leak test Quality Assurance b Reactivate catalyst regularly i 0 5 1 hours at 160 C 100 C c Take care that no dust comes into the unused Jar tube s 2 Mart Anaerobic Jar a Grease the O ring regularly b Grease the coupling 4 times per year with teflon spray c Do not over tighten stainless steel jar clamp i TIGHT TIGHT d Do not disinfect the Mart Anaerobic Jars with alcohol i Use disinfectant provided by Mart Microbiology B V 3 General a Do not switch the Anoxomat on and off within a few seconds b Install the gas cylinder s a
17. ccording to local standards and regulations For further information contact your local dealer Mart Microbiology BV Page 10 17 9 2004 Copyright Mart User Manual Test the equipment The gas supply is opened and by means of the reducing valve the operational pressure should be reduced to 1 8 bar Caution Too much deviation from the required operational pressure may cause malfunctioning or even damage the system After the electrical cables the vacuum gas tubes the frequency 50 60 Hz and voltage 220 230V resp 110V marked at the back are checked the system can be switched on with the switch next to the power plug at the backside The Anoxomat self test program performs a test on the functioning of the individual Anoxomat system components which appear to the user as follows The vacuum pump inlet valve and the jar valves are opened and closed clicking of the valves The pump is tested The pump will run a few seconds The light emitting diodes LED on the indication panel illuminate System is calibrating to the actual atmospheric pressure An acoustic signal indicates that the Anoxomat is in the operational mode and the display only WS8000 WS8080 and WS9000 shows the actual atmospheric pressure and the Anoxomat type number The Anoxomat self test program takes about 15 seconds Caution Do not switch off the system during the Anoxomat self test program Principle of operation In this paragraph t
18. e micro aerophilic recipe Instead of evacuating 70 of the jar volume in the anaerobic evacuation phase 80 of the jar volume is evacuated The standard anaerobic recipe repeats this evacuation and replacement cycle 3 times as is described in several scientific publications P Initially there is approximately 20 oxygen in the atmosphere in the jar the same percentage as normally in the air After every evacuation cycle only 20 of the original volume in the jar is left There is no O in the replaced gas mixture so after the three replacement cycles there is 0 2 X 20 0 16 O in the jar The catalyst takes this very small percentage of oxygen remaining in the jar away which should be included in the jar with the anaerobic cycle Quality assurance The integrated Quality Assurance program assures a desired jar atmosphere in advance In bacterial cultivation the key is that the cultivation environment is guaranteed and reproducible To assure this the following primary aspects have to be certain e airtight jars e appropriate gas supply e sufficient catalyst activity With the standard micro aerophilic and anaerobic program and with programmed recipes including a quality test the quality assurance test is selected by pushing the Quality assurance button on the operator panel The quality assurance is selected before one of the standard recipes is started Mart Microbiology BV Page 13 17 9 2004 Copyright Mart User Manual
19. e volume is evacuated and the light emitting diodes extinguish one by one The light emitting diode row indicator is a measure for the volume evacuated from the jar i e if only one half of the light emitting diodes illuminate 50 of the jar volume is evacuated In the replacement phase the light emitting diodes indicate that the anaerobic gas is injected Quality assurance light emitting diodes The quality assurance light emitting diodes show the operator whether all conditions for successful bacterial cultivation are fulfilled or not The processing unit performs a range of tests to guarantee these conditions The green light emitting diode Passed Fig 3 will illuminate when all quality assurance tests are passed successfully When one of the conditions failed to pass one of the tests the Anoxomat rejects the anaerobic jar and a red light emitting diode will illuminate to indicate the failure cause The selected recipe will not continue with this jar but will proceed with the other jar s If the other jars are processed correctly they can be disconnected and incubated The erroneous jar can be processed again after the Reset button on the operator panel is pushed In this way the user is obliged to find the cause of failure Accidentally continuation after failure is not possible Mart Microbiology BV Page 15 17 9 2004 Copyright Mart User Manual The quality assurance tests are split into three groups e jar leak test e
20. ers of the MART Anoxomat system models WS80 WS8000 WS8080 and WS9000 We are happy to inform the user about the general characteristics and the specifications of the system the installation and the principle of operation are also described The principles of programming the WS8000 WS8080 and WS9000 are described in the paragraph about the programming panel Mart Microbiology BV PO Box 165 7130 AD Lichtenvoorde The Netherlands Tel 31 544 396688 Fax 31 544 396611 E mail info anoxomat com Internet http www anoxomat com Mart Microbiology BV Page 5 17 9 2004 Copyright Mart User Manual Configuration of the Anoxomat system Anoxomat system Figure 1 Anoxomat system WS8080 Anoxomat WS Model Mart Jars Vacuum pump Gases Reducing valves 1 8 bar oP ON Depending on the model 1 up to 3 gas cylinders may be used and 1 upto 3 jars may be used Mart Microbiology BV Page 6 17 9 2004 Copyright Mart User Manual Specifications Anoxomat characteristics Mart Microbiology B V produces 4 different systems for specific use All models incorporate the complete quality assurance programs and the standard basic Micro aerophilic and Anaerobic recipe Table 1 Characteristics EEE o WS8000 WS8080 WS9000 1 3 2 1 CO CO z ar No Yes Yes Yes Yes 1 8 bar 1 8 bar 1 8 bar 1 8 bar Micro aerophilic Micro aerophilic Micro aerophilic Micro aerophilic Anaerobic Anaerobic Anaerobic
21. ertain level e one item is always Back leading to the previous level item e selection item chapter which is always on the display of the programming panel e by pushing the DO button an item is selected e with Back a level menu is selected for other items on a lower level e atthe lowest level DO assigns a parameter Changing the number of evacuation replacement cycles The programming features can be demonstrated by for example changing the number of evacuation replacement cycles of the standard anaerobic program Table 6 experiment change the cycles of the anaerobic recipe ERS Program mode Until item Anaerobic Cycle fo Display Displays numbers of cycles selected Until item mod In the next steps the number of anaerobic cycles is modified until the required number of cycles is displayed on screen number of cycles is selected and saved TESK Operational mode Once the operational mode is entered again the standard anaerobic program will perform the new selected number of evacuation and replacement cycles This example is useful to experiment with the Anoxomat programming display However we advise to leave the standard anaerobic recipe unchanged and to program a recipe as described in further on Mart Microbiology BV Page 19 17 9 2004 Copyright Mart User Manual Changing the display intensity The intensity of the characters on the liquid crystal display can be changed via the programming
22. gas input test e positive or negative catalyst test In next paragraphs the quality assurance tests will be discussed in more detail Airtight There are diverse types of leaks in jars e Very fine cracks thin like a hair and hardly visual e A fixed particle on the seal between jar and lid e Incase of intensive improper usage A jar lid might not be fully smooth or the jar ring could be damaged dry or cracked e Due to improper maintenance of the coupling on the jarlid the coupling may not be airtight when the jar tube is disconnected Some leaks only appear in case of overpressure and others only in case of underpressure All kinds of leaks even the very small ones influence the reproducibility of bacterial cultivation and are unacceptable The Anoxomat has a high precision pressure sensor and with a special algorithm implemented in the processing unit s software a wide range of leak types will be detected When the standard anaerobic or standard micro aerophilic recipe is selected no leak test is executed However it is advised to run the quality assurance test regularly to test the jars extensively Many publishers emphasize the importance of the total absence of Oxy en in relation to survival and growth of anaerobic bacteria as is for example quoted in a reference a publication The rapid achievement and maintenance of a low oxygen tension or the absence of oxygen is an essential requirement for the successful cultivat
23. he principle of operation of the Anoxomat will be explained The main system components of the Anoxomat are User interface Processing unit Valve and pressure system Electronics to control the valve system and vacuum pump Electronics to control the display and sensor signals At the user interface three different areas can be distinguished which will be described successively Operator panel e Indicator panel e Programming panel not on WS80 Mart Microbiology BV Page 11 17 9 2004 Copyright Mart User Manual Operator Panel The user who uses the Anoxomat in daily laboratory routine connects one or more jars and selects the required program on the operator panel to start one of the standard recipes After the program is completed program ready signal sounds the jars can be disconnected and stored into an incubator Reset O Quality assurance O s Anaerobic Cycle Microaerophilic Cycle Figure 2 Operator Panel A yellow light emitting diode on the right side of the push button illuminates if the user pushes the button on the operator panel Another possibility is to start a user defined recipe by pushing the DO button on the programming panel The actual selected recipe is executed How to change the actual recipe is described in the paragraph Selection of a recipe in this manual Activation of the recipe with the DO button instead of the standard anaerobic or micro aerophilic button is done
24. incurred during shipment EXCEPT FOR THE EXPRESS WARRANTY STATED ABOVE MART MICROBIOLOGY B V MAKES NO OTHER WARRANTIES WHETHER EXPRESS OR IMPLIED WITH RESPECT TO THIS MART MICROBIOLOGY B V PRODUCT ALL IMPLIED WARRANTIES INCLUDING THOSE OF MERCHANT ABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE HEREBY LIMITED TO THE DURATION OF THE EXPRESS WARRANTY STATED ABOVE The limited warranty stated above constitutes the sole and exclusive remedy for the original consumer purchaser with respect to any defective MART MICROBIOLOGY B V products and is in lieu of all other obligations or liabilities of MART MICROBIOLOGY B V In no event shall MART MICROBIOLOGY B V be responsible for any costs of procurement of substitute goods loss of profits or any incidental consequential and or special damages of any kind resulting from breach of any applicable express or implied warranty breach of any obligation arising from breach of warranty or otherwise with respect to the manufacture and sale of any MART MICROBIOLOGY B V product whether or not MART MICROBIOLOGY B V has been advised of the possibility of such loss or damage OUT OF WARRANTY REPAIR If your MART MICROBIOLOGY B V product requires service other than under warranty please contact MART MICROBIOLOGY B V for available repair information Every reasonable effort has been made to ensure that MART product manuals and promotional materials accurately describe MART MICROB
25. ion of anaerobics For the WS8000 and WS8080 the test is applied successively for each connected jar and for the WS80 and WS9000 for one single jar If one or more of the jars has a leak the matching light emitting diode illuminates on the indication panel Gas input In the second part of the quality assurance test the availability of the replacement gas is tested A red LED will show if the pressure supplied by the reducing valve of the cylinder to the Anoxomat input is lower or higher than 1 8 Bar WS8000 LEDS 1 2 and 3 will respond to the respective jar tubes WS8080 LED 1 responds to gas connection no 1 and LED 2 responds to gas connection no 2 only in case the 2 gasses are actually used With standard recipes anaerobic and microaerophilic LED 2 will not show any response WS9000 LED 1 responds to gas connection no 1 LED 2 responds to gas connection no 2 and LED 3 responds to gas connection no 3 only in case the 3 gasses are actually used With standard recipes anaerobic and micro aerophilic LED 2 and 3 will not show any response Mart Microbiology BV Page 16 17 9 2004 Copyright Mart User Manual Catalyst activity To create an anaerobic atmosphere there have to be palladium coated aluminium pellets in a steel gauge sachet in the clip of the jar lid to serve as a catalyst The MART Catalyst is a cold catalyst The function of the catalyst is to stimulate the reaction of Hz with O to HO Media if they are
26. ivered separately with the machine This card describes the installation of the MART Anoxomat system in detail Installing the Anoxomat system is only a matter of minutes when all components are available Mart Microbiology BV Page 8 17 9 2004 Copyright Mart User Manual MART ANOXOMAT SYSTEM INSTRUCTION CARD Anoxomat system b the Anoxomat and the gas cylinder Fasten the tubes with the tube clamps 4 Open the gas supply on the gas cylinder 5 Set the gas pressure with the reducing valve on 1 8 bar precisely 6 Switch on the Anoxomat 7 After about 10 15 seconds the red and yellow LED s on the Anoxomat switch out Start up time 8 The Anoxomat is ready for use 9 Connect one or more jars with the jars tubes a Pull the outer ring of the valve on the jar down and push in the black jar tube b To disconnect hold the jar tube and pull down the outer ring of the jar valve Front Panel Anoxomat WS Model Mart Jars Vacuum pump Gases _Reducing valves 1 8 bar Depending on the model 1 upto 3 gas cylinders may be used and 1 upto 3 jars may be used nap QUALITY ASSURANCE 1 2 3 os Airtight os Gas input Connections Catalyst activity Passed 1 Jar a LED s monitoring system pressure i Every green LED is 100 mbar b Quality assurance level i LED 1 is level one etc 2 Quality assurance 1 2 3 i Jar indication ii Gas supply indication a
27. k K ex nos 8 ON 8 Table 4 gt MIXZAS COMPOSTION sissi ideation ta su aa bean aia a lanas HhHhHFFHNHRaRaXaXa aRaXRXKXreeeem 8 INIA 5 5k43 33 4 aie 4 k b e d da 2 sorsero ke d d Qas 5 3 ke d a 5 2 ike d d 8 a ie d d iosas ke d s d k a5344 4E e ss 8 TEST THE EQUTPMEN TT g4 x xia Sak Seke d discs 11 PRINCIPLE OF OPERATION aci 11 OPERATOR PANEL AAA O 12 Figure 2 Operator Panel iii di irte Hide la G rey lan tse 12 O NN 12 STANDARD MICRO AEROPHILIC RECIPE eke keke keke keke keke ke keke kek keke kek ke ka kek KK KK K KE K KA KA KA KE KA KA KA KA KA 12 CALCULATION REST PERCENTAGE OXYGEN IN STANDARD MICRO AEROPHILIC RECIPE oooooonccnccccoonnonanoncnnononnnnos 13 STANDARD ANAEROBIC RECIPES a S R i ce yV vw atc ieee abbas 13 QUALITY ASSURANCE Anas 13 Table 5 Levels of quality ASSUVANCE cccccecessceenseenseceeneeessecesceecsseceeaeecsseceeceecsaecseceecsaecsneecsaeceeneecsaeceeeeeeaeens 14 INIA NANO 15 Figured Indicator panela anida 15 LIGHT EMITTING DIODE BAR JAR oooncccnnncncncnnnnonononononononononononononononononononononononononononononononononononononononinenenenenenoss 15 QUALITY ASSURANCE LIGHT EMITTING DIODES uu ekere keke e k keke ke ke kek keke ke k K Ke Kake KA KA KA ka ke k kk k Kr kek 15 ARTIGAT anan dde 16 GAS INPUT di 16 CATALYS TACTO Y A dd a a a a area t a 17 PROGRAMMING PANEL
28. on in the air is initially 20 and should be reduced to 5 a factor 4 times as low in the evacuation cycle 5 Low set point 20 x 1000 250 There has to be 10 CO in the gas composition During a range of 100 mbar CO has to be injected a4 00 CO dosing range 10 100 High set point 1000 Intermediate set point High set point dosing range CO 1000 100 900 Mart Microbiology BV Page 23 17 9 2004 Copyright Mart User Manual For accuracy reasons it is better to have the long ranges in the deepest vacuum points The explanation is straightforward Injecting gas means physically that a valve is opened and is closed again as soon as a certain pressure setpoint is reached If there is a deep vacuum in the jar gas is injected into the jar with a high speed The lower the vacuum in the jar the easier the gas flow can be better controlled and the more accurate the end result will be Program description and programming Similar to the previous program activities the recipe table has to be programmed Table 13 Programming steps for recipe 17 with 5 Oz 10 CO and 85 N2 Program level Item input lt Program recipe gt lt recipenumber gt lt 17 gt lt Program recipe gt lt actionnumber gt lt 1 gt lt Program recipe gt lt mod Action gt lt 1 gt lt Program recipe gt lt mod gas gt lt 0 gt lt Program recipe gt lt mod Setpoint gt lt 250 gt lt Program recipe gt lt action number gt l
29. r Displa EGEE E x E Display format Display Back Display Back Calibrate ext Are you sure a E El 2 o Sl F3 BIS cle ojo E Fi Reference val 2 Back Reference val 1 Mod Back Reference jar Offset BES airy Back Program system Anoxomat type al UC el Load r Do not do Program settings Catalyst margin Displa m D f E Leaktest margin Display Back Highest airpress Display Use arrow up only here L______ r Highacc Jar sel Lowest airpress Displ oy gt E z E E Lowacc jar sel zjm FIR K Displa Di Waittime jar sel play Back Display Correction mode Select recipe gt Anaerobic cycle Back Mart Microbiology BV Page 18 17 9 2004 Copyright Mart User Manual The Anoxomat systems with a programming panel have two modes the operational mode and the program mode With the Esc button you can switch from the program mode to the operational mode In the previous paragraphs the Anoxomat was assumed to be working in the operational mode Terms used for the program mode are menu e list with items to select from e pushing the T button displays the previous item circular and the 4 the next item level e each menu contains items on a c
30. s saving time In case of older but reactivated catalysts a longer testing period may be necessary It is necessary to order new catalysts In user defined programmed recipes the quality assurance functions can also be included In this case the desired quality assurance level has to be activated as well on the operator panel before starting the user defined recipe For including quality assurance functions in a user defined recipe see paragraph programming recipes Mart Microbiology BV Page 14 17 9 2004 Copyright Mart User Manual Indicator Panel E QUALITY ASSURANCE 1 2 cs Airtight os Gas input of Catalyst activity 2 a Passed O m Figure 3 Indicator panel Green light emitting diodes illuminate when all conditions are satisfied In alarm situations the red light emitting diodes illuminate When all quality assurance tests are passed the light emitting diode Passed will illuminate Light emitting diode bar JAR The light emitting diode bar JAR has 2 main functions e Indicates the stage of the evacuation and replacement cycle e Indicates the level of quality assurance All light emitting diodes in the vertical row under the word JAR will illuminate when either the standard micro aerophilic recipe or the standard anaerobic recipe is executed When all light emitting diodes are on the pressure in the jar s is atmospheric In the evacuation phase th
31. ssities to maintain all jars working with the Anoxomat original Mart jars as well as modified jars of other brands is available This kit includes 10 replacement snap shut couplings these cannot be cleaned only be greased with Teflon spray tools cleanser and lubricant complete with instructions Also available is a refill set consisting of only the used materials of the toolkit Mart Microbiology BV Page 27 17 9 2004 Copyright Mart User Manual Refences Fildes P Macintosh J An improved form of anaerobic jar Br J Exp Pathol 1921 2 153 Brazier J S and Smith S A 1989 Evaluation of the Anoxomat a new technique for anaerobic and microaerophilic clinical bacteriology J Clin Pathol 42 640 644 W Klietmann K N sner J Focht S Heinrich A New concept for the cultivation of anaerobic bacteria Koneman EW e a Diagnostic Microbiology 3rd edition 1988 1989 Ch 11 396 J B Lippincott Company Philadelphia Sutter V L Citron D M Edelstein M A C Finegold S M Wadsworth Anaerobic Bacteriology Manual 4th Edn Summanen P Baron E J Citron D M Strong C A Wexler H M and Finegold S M 1993 Wadsworth Anaerobic Bacteriology Manual 5th Edn Star Pulishing Company Co Belmont CA Imhof A and Heinzer 1996 Continuous monitoring of oxygen concentrations in several systems for cultivation of anaerobic bacteria Clin Microbiol 34 1646 1648 Mart Microbiology BV Page 28 17 9 2004 Copyright
32. t 2 gt lt Program recipe gt lt mod Action gt lt 3 gt lt Program recipe gt lt mod gas gt lt 0 gt lt Program recipe gt lt mod Setpoint gt lt 0 gt lt Program recipe gt lt actionnumber gt lt 3 gt lt Program recipe gt lt mod Action gt lt 2 gt lt Program recipe gt lt mod gas gt lt 1 gt lt Program recipe gt lt mod Setpoint gt lt 900 gt lt Program recipe gt lt action number gt lt 4 gt lt Program recipe gt lt mod Action gt lt 2 gt lt Program recipe gt lt mod gas gt lt 1 gt lt Program recipe gt lt mod Setpoint gt lt 1000 gt lt Program recipe gt lt action number gt lt 5 gt lt Program recipe gt lt mod Action gt lt 0 gt lt Program recipe gt lt mod gas gt lt 0 gt lt Program recipe gt lt mod Setpoint gt lt 0 gt Esc Operational mode ca Correction Mode Before a newly programmed recipe can be used it has to be executed with the Anoxomat system in the correction mode The Anoxomat system calculates the settings for the new defined program Caution When a new programmed recipe is not executed in the correction mode the expected composition will differ from the real one The programming steps are Table 14 Correction mode hd program mode lt correction mode gt lt on gt nad operational mode start recipe 17 in correction mode After the recipe is executed in the correction mode the correction mode returns a
33. this section the activities are described which have to be done to add a recipe to or replace a recipe in the Anoxomat system database Next list summarizes the actions which should be performed subsequently definition of the gas composition study the recipe parameters determine how the defined gas composition can be made construct recipe table calculation of the setpoints program description and programming Mart Microbiology BV Page 21 17 9 2004 Copyright Mart User Manual select user defined recipe set correction mode on run the recipe in the correction mode repeat last 2 steps two to three times In the next paragraphs the programming of a recipe on a WS9000 system is described Gas 1 is Anaerobic gas mixture not used in this example The two other gasses are separately connected to the WS9000 Definition of the gas composition Consider the establishment of a low oxygen concentration in a jar The gas composition required could be 5 O 10 CO 85 Na Recipe parameters A recipe consists of a sequence of actions each with the attributes Table 10 Parameters Description Input The action number defines when the Actions are numbered from 1 to 25 action is performed in the sequence the lines of the recipe Which action has to be taken 0 no action 1 evacuation 2 replacement 3 vacuumleaktest 4 catalyst test positive anaerobic 5 catalyst test negative micro aerophilic Which gas has to
34. utomatically to the off state This is done to prevent the customer from running a recipe in the correction mode always Mart Microbiology BV Page 24 17 9 2004 Copyright Mart User Manual In certain cases when setpoints lie close to each other running the recipe in the correction mode one or two times more can increase the accuracy Caution Recipe 17 can be run with or without a leaktest This depends on whether the quality assurance is switched on or not Programming a recipe Table 15 Programming a recipe EN PROGRAM MODE until Menu Select Menu until Program Recipe Select Program Recipe until Recipe number on display Recipe until right number on display Recipe number until Action number on display Action until right Action number on display Action number until Mod action on display Mod until right Action i on display Mod action until Mod gas no on display Mod i until right gas selection on display Mod gas no until Mod setpoint on display Mod until right setpoint number on display Mod setpoint until Read action on display current Recipe number Action number Mod action number Gas number setpoint on display Read action With esc exit PROGRAM MODE until Back on display Program recipe until Back Do on display Menu esc exit PROGRAM MODE Mart Microbiology
35. vacuation phase of the standard micro aerophilic recipe of all Anoxomat models 70 of the jar volume is evacuated The Anoxomat achieves this with high accuracy due to the high precision pressure sensor Assuming the atmospheric pressure is 1030 mbar the standard micro aerophilic recipe evacuates until 0 3 1030 309 mbar In the successive replacement phase the jar is filled with an oxygen free gas mixture Initially there is approximately 20 oxygen in the atmosphere in the jar the same percentage as normally in the air At the end of the evacuation cycle only 30 of the original gas volume in the jar is left There is no oxygen in the replacing gas mixture so after the replacement cycle there is 0 3 x 20 6 Oz in the jar With the programmable Anoxomat models recipes for a reduced oxygen concentration atmosphere can be programmed easily as described in the paragraph Programming recipes A similar calculation is required before programming Assume that a reduced oxygen atmosphere with 8 oxygen is required To obtain this atmosphere 8 20 40 of the oxygen must remain in the jar so 60 of the jar volume should be evacuated The necessary low set point for this recipe can be calculated directly and entered into the program 1030 0 4 set point 412mbar Standard anaerobic recipe The standard anaerobic recipe performs three evacuation and replacement cycles which are different from the one evacuation and replacement cycle of th
36. when there are specific requirements for the jar atmosphere In subsequent paragraphs the following operator panel functions will be described briefly e Reset the Anoxomat e Start standard micro aerophilic recipe e Start standard anaerobic recipe e Select the quality assurance level Reset The main purpose of the reset button is to prevent errors Users are made aware of erroneous situations and are forced to take the action required Before another jar can be processed the reset button has to be pushed first A signal sounds when other switches are pushed Standard Micro aerophilic recipe The standard micro aerophilic recipe has one evacuation and replacement cycle which is composed of two phases e evacuation phase e replacement phase The standard micro aerophilic recipe composes optimum breeding results for micro aerophilic organisms such as for instance Campylobacter jejuni Mart Microbiology BV Page 12 17 9 2004 Copyright Mart User Manual For WS80 and WS8000 users it could be of interest that J S Brazier proved good growth conditions for capnophilic bacteria Neisseria Heamophilus en Gardnerella in the jars which were processed with the standard micro aerophilic recipe The users of the WS8080 and WS9000 have the possibility to create micro aerophilic atmospheres in a jar with various O2 CO and N concentrations Calculation rest percentage oxygen in standard micro aerophilic recipe In the first e
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