CONTENTS NEWS WORLD DIAGNOSTICS DIAGNOSTICS CLOSE UP STAY HEALTHY KNOWING MORE

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2 CONTENTS NEWS Another company acquisition by Cormay Chemistry days Our Participation in Clinical Lab Expo 2011 in Atlanta WORLD DIAGNOSTICS Critical Evaluation of Faecal Parasite Concentrators 6 10 DIAGNOSTICS CLOSE UP Evaluation of Parasep Faecal Parasite Concentrator A Comparison of Traditional Methods and Parasep STAY HEALTHY Unwanted Souvenirs KNOWING MORE A Concise Atlas of Parasites Publisher: PZ Cormay SA ul. Wiosenna Łomianki tel.: faks: office@cormay.pl Editing: Chief Editor Monika Dziachan PZ Cormay SA Editing and proofreading Agape Preparation and production: Agape. Consultants and editors ul. Lazurowa 183 lok. 3, Warszawa tel./faks: biuro@agape.com.pl, The editor reserves the right to shorten and edit published materials. 2 Whole truth in one drop CORMAY International Bulletin

3 EDITORIAL Children s Infections T hey set up their own family like humans. Paresites like family life not in the mountains or at the seaside but in our bodies. Our children are most prone to parasitic infections, as they do not always obey the hygiene rules. They will pet a passing dog or cat, play in the sand and swim in a pool or a lake, unaware of the risk. They eat fruits from trees and vegetables from vegetable beds. That is why they are constantly at risk of infections by giardia lamblia, toxoplasma gondii, pinworm, toxocara or roundworm. The statistics are alarming. Over 50% of kindergarten children are infected with parasites. The test is non-invasive, and the detailed result is available in a few hours Although cysts can be detected by a simple non-invasive stool test, treatment of parasitic infections is still difficult. The test explains the reasons of abdominal discomfort such as constant pain, nausea and general feeling of illness. Stool analysis also helps detect digestive disorders and malabsorption symptoms. We must remember that even if one family member is infected, the whole family must undergo the tests. What is required to obtain a reliable result? A microscope, an experienced diagnostician, suitable transport tubes with a spatula and a built-in filter. In this bulletin you will find a comparison of faecal parasite concentrators. Thanks to Parasep system, the test will be easy and pleasant. The kit prevents exposure to the tested material and an effective filtering system helps obtain a high-quality specimen. The statistics are alarming. Over 50% of kindergarden children are infected with parasites. The test is non-invasive, and the detailed result is ready in a few hours. Treatment is easy and effective; however the complications of untreated parasitic infections can be troublesome. Why does it happen then? You are welcome to read Our Laboratory and to test the Parasep system yourself. Beata Stefañczyk Product Manager Parasitology Why it is worth to have a stool test? Nr 2 (22), summer 2011 Whole truth in one drop 3

4 NEWS Another Company Acquisition by Cormay An interview with Tomasz Tuora, Chairman of the Board, PZ Cormay SA. What is the driving force behind the company s rapid expansion? Tomasz Tuora: There are several factors, mainly the investors financial support and EU funding. We also met the right people at the right time the scientists whose new ideas we decided to support. Let s start from the beginning the investors, whose support made it possible to acquire the Irish company called Innovation Enterprise. T. T.: The acquisition is very recent, it was finalised on 19th July. We became interested in the Innovation Enterprise, because it owns numerous test production technologies which we did not possess over 220 tests performed by various methods in total. They are also very experienced when it comes to point-of-care tests. One of the deciding factors was the fact that we would be able to broaden our portfolio. Furthermore, we are now able to geographically diversify our sales. Innovation Enterprise owns a company in China and accomplishes 40% of sales in China and India. It is a crucial factor, because by 2018 China may become our largest market. Thanks to the acquisition we will also begin to supply the largest companies. One of Innovation Enterprise s clients is Beckman. Furthermore, such diversified technology is driven by a highly talented scientific research team. It is especially important when it comes to commercialisation of a global revolutionary diagnostic test technology. The scientists are another important factor. T. T.: Together we work on technologies and a blood analyser which may cause global changes in the field of medical diagnostics. The product will be ready in The prototype will be available at the beginning of next year. Right now we are at the industrial studies stage. Afterwards we need to perform commercialisation and industrialisation. The new Cormay product will serve as biochemical, immunological and molecular blood analyser. It will perform three tests per second which translates into up to 10 thousand tests per hour. Analysers currently available on the market perform up to 2 thousand tests per hour. One drop of blood will be enough to perform 100 tests. Another novelty will be the analyser s compact size. All that thanks to Polish scientists? T. T.: I am afraid to think that their drawers are still full of projects awaiting implementation, which may never happen due to lack of funds, for example. Let s be optimistic, though. Thanks to companies like ours the scientists can put their ideas into practice the more the better. TOMASZ TUORA FACTS A 34-year-old graduate of Economics Department of the University of Warsaw and University of Miami. In PZ CORMAY SA for 10 years. Since 2006 Tuora has been holding the position of the Chairman of the Board. He initiated and carried through the company debut on the Warsaw Stock Exchange as well as acquisitions of Swiss company Orphee SA. and Irish Innovation Enterprise. Enterprise owns a company in China and accomplishes 40% of sales in China and India. It is a crucial factor, because by 2018 China may become our largest market novative Economy, such as elaboration of own analysers and reagents. These funds accelerated execution of our plans, which seemed long- -range at the time. Now everything becomes real. With such rapid growth we must invest in development of our staff. A training programme prepared for our employees aims at development and improvement of their managerial, communication and sales skills. This project is also supported by the EU. All our actions are the result of implementation of our development- -oriented strategy, broadening our offer and improving our standard of service. We are very proud that, being a Polish company, we develop according to global standards. Another factor that you mentioned was EU funding. T. T.: We are currently implementing five projects co-funded by European Regional Development Fund within Operational Programme In- Thank you. Interviewed by Monika Dziachan 4 Whole truth in one drop CORMAY International Bulletin

5 Chemistry Days We will make your idea possible was the slogan that promoted the stand of PZ Cormay SA during Chemistry Days in Warsaw on 2 nd 3 rd June. NEWS The goal of our participation in this event was to begin the co-operation with faculty and graduates from various chemistry departments. The event was successful, we made several useful contacts which may influence the development of new technologies in medicine says Dr Renata Filipek, Research and Development Department Specialist, PZ Cormay SA is international year of chemistry, and also a Marie Curie year in Poland. It was named three years ago by the United Nations upon the request of UNESCO and International Union of Pure and Applied Chemistry in order to draw the public attention to chemistry s contribution to human development. Without chemistry we would not exist. Therefore, a good knowledge of chemistry phenomena is crucial and it determines the quality of our lives and contributes to the development of medicine adds Renata Filipek. For two days not only did we hold numerous meetings with students and graduates but also had a pleasure to meet Professor Maciej ylicz, MD, PhD, the President of Foundation for Polish Science and exchange opinions on current cancer treatment methods with him. Celebrations of International Year of Chemistry 2011 are under honourable patronage of Poland s president, Mr Bronis³aw Komorowski. For two days we held meetings with students and graduates It was a fruitful event and we established a number of important contacts Our Participation in MEDICA 2011 in Düsseldorf Novelties, novelties, novelties these words summarise Cormay Group s participation in international Medica trade fair in Düsseldorf. Three times larger stand, presentation of a biochemistry analyser prototype, and launch of the new logo are only some of them. The prototype of own biochemistry analyser EQISSE with throughput of over 400 tests per hour attracted the most interest. Official launch of the new analyser is said to take place in 18 months. End users were especially impressed with an unlimited number of sample positions, extra rotor for QC samples, calibrators and STAT samples and a unique solution for liquid waste condensation. Analyser construction specialists were impressed with simplicity of service and maintenance says Wojciech Przybecki, Project Manager. We would like to cordially invite you to the future launch of the new analyser. As a supplement of our portfolio we also presented a new semi-automatic biochemistry analyser Mutli, which still attracts a lot of interest on emerging markets. During the four-day fair we met the majority of our distributors from all over the world. Most of them were surprised by our rapid development, some of them admitted that this was what they expected says Pawe³ Mirosz, Export Manager PZ CORMAY S.A. There are also new possibilities of cooperation with leading companies in our industry. We are very pleased with confidence which is placed in us he adds. This year for the first time the companies associated in Cormay Capital Group presented their products with a new collective logo. During the fairs we were looking for suitable investors for our products The conference was also attended by a group of scientists working on commercialisation of an innovative invention Nr 2 (22), summer 2011 Whole truth in one drop 5

6 WORLD DIAGNOSTICS A Critical Evaluation of Commercial Faecal Parasite Concentrators Six commercially available faecal parasite concentrator devices were critically evaluated. All products were compared to the conventional Ridley-Allen Sieve method under identical conditions. The evaluation was primarily based on packaging, clear and concise instructions, ease of use and the quality of the separation of the sample during the filtration. The concept behind most of these commercial products centres on ease of use, with the aim of preventing operator exposure. Filter-unit leakage was observed once filtration had commenced and also when the device was disconnected in three of the devices. The sealing mechanism of the Parasep range prevented any leakage. In three of the products tested there was blockage of the filter resulting in poor filtration and separation. The Parasep devices did not become blocked at any time. Allison Baxendine BSc., MS.c Applied Parasitology & Medical Entomology The role of this evaluation was to take a non-biasedview of the various commercially available faecal parasite concentrators, including the new Maxi Parasep non-centrifugal concentrator from DiaSys Europe ltd. Six commercially available concentrator devices were critically evaluated on the bases of packaging, protocol, physical parameters, ease of use and thequality of separation. The samples were tested for recovery with stool samples seeded with Helminth Ova (Ascaris lumbricoides). The Six commercial products that were evaluated were; FPC Faecal Parasite Concentrator and the FPC Jumbo Faecal Parasite Concentrator (Evergreen Scientific, USA), the Para-Pak Macro-Con Stool Concentration System (Meridian Diagnostics, Inc, USA), Mini Parasep (DiaSys Europe) Parasep (DiaSys Europe) and the Maxi Parasep Concentrator (DiaSys Europe). METHOD The testing procedure was carried out as instructed by each of the manufacturer s on appropriate sample material following the procedure through the filtration process and stopping before the centrifugation step. All products were compared to the Recommended Ridley-Allen (Ritchie) Sieve method. For each of the 6 commercial products 12 devices were tested under identical conditions ensuring that no externalconditions influenced the testing procedure. 6 Whole truth in one drop CORMAY International Bulletin

7 WORLD DIAGNOSTICS FPC FAECAL PARASITE CONCENTRATOR, EVERGREEN SCIENTIFIC, INC, USA The packaging of the product is simple and neat, containing everything that is required to carry out the procedure from start to finish. Each case contains material for 100 tests. The instructions are well presented, explaining ingreat detail the background to the product, methodology and specifications. It is professionallooking, but on reading further the procedure becomes very complicated, confusing the process that is required for fresh and preserved stool samples, giving additional information that is noteasy to follow. In total the procedure contains 16 steps. Once passed the additional procedure information forthe fresh stool, steps 1 10 are clear and easy to carry out. It is difficult to transfer the sample from the faecal transport tube into the flat bottomed tube containing the formalin as the neck of the tube is very small and the sample very easily attaches to themouth and sides of the tube. At steps 7 & 8 there is sample leakage when attached together; it leaks from both sides of the strainer unit when inverted. Once the device is inverted there is immediate blockage of the filter, allowing very little sample to filter through, even if tapped lightly. Out of the 12 devices tested ¾ of the sample was left unfiltered in 8 of the devices. The other 4 devices filtered most of the sample over a period of seconds. The filter became blocked very quickly and required a great deal of gently tapping to dislodge the blockage to allow enough of the sample to filter through for a satisfactory end sample. If there were no vent-straw very little sample would filter through due to a complete blockage of the sieve with no chance of an equilibrium of pressure being reached on either side of the filter matrix. At steps 7 & 8 there is sample leakage when attached together; it leaks from both sides of the strainer unit when inverted. Step 9 requires unscrewing of the FPC strainer with the flat bottomed tube still attached from the centrifuge tube, this is not possible without the sample leaking all over your hands. This occurred on 11 out of the 12 devices that were tested. From Step 1 10 the test took approximately minutes to complete. General observations: Using a 15ml flat bottomed tube to add the sample, 9 ml of formalin, 3 ml of ethyl acetate and 8 drops of Triton X-100 there is very little space left in the tube. This may have exacerbated the amount of leakage that was observed simply due to too an excess of material in the tubes. There is no regularity in the filtration rate through the strainer unit, once the filter became blocked there is very little chance of unblocking it to allow for a satisfactory amount of the sample to be filtered and separated. Even if left to allow the pressure to become equilibrated there is no further filtration of the sample. The formation of a layer on the filter matrix also causes secondary filtration, and hence a reduction in the number of parasites passing through the filter. This is reflected in a comparatively low recovery rate.. MAXI PARASEP CONCENTRATOR: The ability to fit most transfer vials directly onto the filter means that this device is excellent for the processing of remote or field samples FPC JUMBO FAECAL PARASITE CONCENTRATOR, EVERGREEN SCIENTIFIC, INC, USA Packaging of the product: very simple but neat containing everything that is required to carry out the procedure from start to finish. Each case contained materials for 120 tests. The instructions are well presented, explaining in great detail the background to the product, methodology and specifications. It is very professional looking, on reading further the procedure becomes very complicated, confusing the processes that are required for fresh and preserved stool samples. When using preserved samples it does not indicate how much of the sample should be used. The instructions for fresh samples are also not clear. For fresh liquid stool there are an excess of 30 ml vials, used for the preserving steps. If this device and procedure was designed to cut down on the operator exposure to hazardous substances the initial procedure seemed to increase exposure. Is it really necessary to leave fresh stool sample in formalin for 30 minutes to allow for preservation of the organisms? There seems to be no explanation as to the time scale of this statement. The procedure only details how much fresh stool sample should be used in the process (2 spoonfuls for solid stool and 3 spoonfuls of fresh liquid stools). If the lab is sending out collection vials to enable the stools to return to labs already preserved there must be instruction as to how much stool is used as it cannot be expected that every patient will fill them correctly. There is no instruction explaining what happens with the vials that come into the labs already filled with the faecal sample and correctly preserved. Steps 1 4 are easy to follow but again as explained for Product 1 there is no clear indication from what point one measures where to pull the vent-straw from. Steps 6 & 7 are clear. During Step 8, after the initial few millimeters of the sample filtered through, the strainer unit became blocked. The instructions in step 8 suggest that if there is blockage lightly tap the centrifuge tube, but this did to make any difference only by tapping the transport vial did any of the sample filter through. After approximately seconds of light tapping and holding the tubes at a slight angle did the sample completely filter through. Blockage of the jumbo strainer was observed in each of the 12 devices tested, but with light tapping the sample was filtered through satisfactorily. Step 9, unscrewing the jumbo strainer with the transport vial attached from the centrifuge tube, there is leakage of the sample from both the jumbo strainer and the centrifuge tube (9 out 12 devices leaked). From steps 1 10 the test time took approximately minutes. General observations were that it was well presented with all of the necessary items provided; though there maybe no requirement to supply applicator sticks, biohazard bags and dispensing spoons, nor the extra transport vials. Using the larger device was easier to handle in terms of adding the sample to the transport vial and there was notably less leakage during filtration and taking the device apart after filtration had taken place. If you are sending the transport vials out to the patients do you send the vials which are part of the package or do you use ones that are already available in the lab and then transfer the sample into the correct transport vials? If they are sent out already pre-filled then the instructions should be aimed at those, omitting the need to add the formalin, ethyl acetate and triton X-100. This did cause some confusion as the general aim of these devices should be to reduce time and operator exposure to the samples by simplifying the procedure. For a simple procedure there are too many steps to before and after centrifugation. It is a prolonged simple test. Nr 2 (22), summer 2011 Whole truth in one drop 7

8 WORLD DIAGNOSTICS PARA-PAK MACRO-CON STOOL CONCENTRATION SYSTEM, MERIDIAN DIAGNOSTICS, INC, USA Steps were carried out as instructed by the manufacturer (the following steps required the use of a centrifuge), enabling a sample to be filtered and separated. The size of the pack does suggest that storage could be a problem. Once the package is opened there is no rack supplied, you cannot have the whole pack sitting on a bench as it is too large. The instructions seem to hold too much information in a small space. It explains the procedure for fresh stool specimens after it described the whole of the specimen processing section. The device is designed principally for stools that come into the lab already preserved, which does cut down on operator time and exposure to the sample. Section A (Specimen collection) does not indicate how much sample is being filtered, it only refers to the Para-Pak package insert. It would be useful to know how much sample is to be filtered. Initial impression of the sealing system between specimen vial and the filtration unit is that it was not a tight fit and there would be excessive leakage. There are too many confusing steps between the attachment of the specimen vial to the filtration unit and the actual filtration of the sample. During the filtration of the sample there is considerable blockage that requires a relatively large amount of forceful tapping of the centrifuge tube on a table top to enable enough of the sample to be filtered through. Eight out of the twelve devices tested only allowed ¾ of the sample to be filtered through. Step 5, requiring you to loosen the filtration unit from the centrifuge tube, simply confuses the process and there is a great deal of material which leaks out, obviously due to the pressure which built up during the filtration. It is also very difficult to only loosen it slightly without taking the whole device apart. There was a certain amount of confusion as to why the device had to be refilled with formalin and ethyl acetate before centrifugation took place. Should the sample that came into the lab already preserved not have been in enough liquid to enable the whole process to be completed without re-suspending the sample. General observations are that the advantage with this device is the reduced sample preparation, due to the sample coming into the lab already preserved in the Para-Pak specimen vial. This simplified the whole process, it is just that once the sample has come into the lab the procedure becomes convoluted. By the time that you had taken the specimen vial and vigorously shaken it for 60 seconds attached the filtration unit to the specimen vial and inverted the device (so that the centrifuge unit is pointed end downwards), the sample had become stuck to the base of the specimen vial again that only the liquid filtered through. You had to find a way of dislodging the sample before you filtered it but after the whole device has been put together. (Steps 1 3). The test procedure from steps 1 8 took approximately 6.5 minutes. MINI PARASEP CONCENTRATOR, DIASYS EUROPE The Mini Parasep is neatly packaged with all requirements for 50 tests. The box requires little storage space. The instructions are schematically represented by four concise and easy to follow diagrams. The protocol allows for a variety of fixatives and solvents to be employed. The integrated nature of the product ensures there are no problems with sample transfer. A three dimensional sieve prevents occlusion and hence all material passes easily through under gravity or centrifugal pressure. Hermetic sealing prevents the leakage of any material. On completion of centrifugation a torque differential allows the base of the device to be removed, and all rejected material contained within the upper unit to be discarded without operator exposure. The supernatant is easily extracted from the base of the device, and the small pore size and high filter area gives a high clarity and parasite load sediment. Other units in the range include reagent prepared devices, allowing operator timesavings. The optional 220 micron secondary filter gives a high clarity sediment and may remove the need for solvent (ethyl acetate/ether). In general this device is simple and quick to use, whilst increasing lab safety by preventing operator exposure. The whole test requires minutes operator time. MIDI PARASEP CONCENTRATOR, DIASYS EUROPE The Parasep is neatly packaged with all requirements for 50 tests. The box requires little storage space. The instructions are schematically represented by four concise and easy to follow diagrams. The protocol allows for a variety of fixatives and solventsto be employed. The integrated nature of the product means that there are no problems with sample transfer. A three dimensional sieve prevents occlusion and hence all material passes easily through under gravity or centrifugal pressure. Hermetic sealing prevents the leakage of any material. Upon completion of centrifugation a torque differential allows the base of the device to beremoved, while all rejected material is contained within the upper unit to be discarded withoutoperator exposure. The supernatant is easily extracted from the base of the device, and the small pore size and high filter area gives a high clarity and a high parasite load sediment. The quantity of sediment is large enough to prepare multiple slides. Other units in the range include reagent prepared devices, allowing operator time savings. The optional 220 micron secondary filter gives a high clarity sediment and may remove the need for solvent (ethyl acetate/ether). In general this device is simple and quick to use, whilst increasing lab safety. The whole test takes minutes (excluding centrifuge time). 8 Whole truth in one drop CORMAY International Bulletin

9 WORLD DIAGNOSTICS MAXI PARASEP CONCENTRATOR, DIASYS EUROPE Our initial impression of the packaging was very simple since only the filtration thimbles were provided. This was due to the fact that the product was designed to incorporate all centrifuge tubes and transport vials. The instructions were very clear. They clearly described the alternative steps that require you to carry out the procedure if the samples arrived into the laboratory already preserved. When using fresh faeces there were 4 easy to follow steps before the centrifugation procedure commenced. If using already preserved faeces there were 2 steps to the filtration process. You simply attach the transport vial to the filter thimble, vortex and invert the device to allow the filtration to commence. Once filtration had commenced there was no blockage of the filter and therefore no requirement to tap or shake the device. When using the smaller 15 ml centrifuge tube the filtration process took between 13 and 15 seconds, whilst using the larger 50 ml centrifuge tube the process took 2 3 seconds. There was complete filtration and no blockage of the filter or leakage from the device. There was approximately 0.5 to 1 ml of fluid left in the transport vial in 3 out of the 12 of the devices tested simply due to the fact that this lay beneath the level of the filter on the filter platform. In general most 30 ml transport tubes fit onto the filter thimble allowing attachment of the preserved sample vials straight away onto the filter thimble. This reduces operator exposure and preparation time. There was no leakage at any time though it did require a relatively large amount of force to ensure a tight fit was achieved between the filtration thimble and the vials. The ability to fit most transfer vials directly onto the filter means that this device is excellent for the processing of remote or field samples. The fact that there is the option of using the larger 50 ml centrifuge tube is excellent if you require to concentrate a larger faecal sample allowing you to use the sample in more than one application. The whole test procedure took approx. 2 3 minutes. Fig. 5. Comparative methods Feature Sieve Method (Ritchie/Ridley Allen) Maxi Parasep Midi Parasep Mini Parasep Size of Mesh (μm) No. Holes Filter Area (mm 2 ) Empirical Recovery Number of Steps Total Test Time (mins) Process Emulsification Centrifugation Reagents (ml) Formalin/ethyl acetate or ether Cleaning Utilised Approx 500 mm 2 utilised >90% 9 10 Open Separate tube Separate tube 7/3 By hand % 4 3 Closed Same tube Same tube 6/2 Disposable ,2% 4 3 Closed Same tube Same tube 6/2 Disposable % 4 3 Closed Same tube Same tube 6/2 Disposable Fig. 5. Comparative methods Feature Sieve Method (Ritchie/Ridley Allen) Evergreen Meridian Macrocon Biosepar Size of Mesh (μm) No. Holes Filter Area (mm 2 ) Empirical Recovery Number of Steps Total Test Time (mins) Process Emulsification Centrifugation Reagents (ml) Formalin/ethyl acetate or ether Cleaning Utilised Approx 500 mm 2 utilised >90% 9 10 Open Separate tube Separate tube 7/3 By hand 600 Standard 600 Jumbo 104 Standard 360 Jumbo 110 Standard 480 Jumbo 82% Standard 87% Jumbo 16 8 Opened to transfer suspension Separate tube Separate tube 9/3 Standard 12/4 Jumbo Disposable % 12 7 Opened to transfer Separate tube Separate tube 12/4 Disposable % 9 6 Open during emulsification Same tube Same tube 3,5/1,2 Disposable Nr 2 (22), summer 2011 Whole truth in one drop 9

10 WORLD DIAGNOSTICS Packaging, clear and concise instructions, ease of use and the quality of the separation of the sample during the filtration are most likely what a laboratory would base their decisions upon on when considering a variety of commercial products. CONCLUSIONS It was decided to base this study in terms of four criteria when testing each of the products. The evaluation was primarily based on packaging, clear and concise instructions, ease of use and the quality of the separation of the sample during the filtration. These four objectives are most likely what a laboratory would base their decisions upon on when considering a variety of commercial products. On the whole the four concentrators were well organized with well presented instructions and all the appropriate material to carry out the procedure efficiently. The FPC Faecal Parasite Concentrator and the FPC Jumbo Faecal Parasite Concentrator (Evergreen Scientific, USA) contained a large number of unnecessary products that could normally be found in an average laboratory. The consideration within a well equipped laboratory would concern needless expense for kit components. It was decided that the Para-Pak Macro- -Con Stool Concentration System (Meridian Diagnostics, Inc, A USA) was very bulky and left little space to store the devices once sitting on the bench before and during usage. This situation could be extremely problematic when dealing with large numbers of samples, or in bustling testing environments. The Maxi Parasep Faecal Parasite Concentrator (DiaSys Europe, UK) looked very basic in the fact that only the single use separation filter thimbles are supplied, but they are essentially all that is required for rapid concentration of intestinal parasites. The filter thimble can be attached onto most 30 ml transport tubes and the samples collected in generic 15 ml and 50 ml centrifuge tubes. The lack of excess packaging and simple design made this product very attractive. Mini Parasep and Parasep are both packaged in small, easy to store boxes requiring little room in the laboratory. They both function in a similar manner to the Maxi-Parasep but are supplied with all components necessary for faecal concentration. Each of the product instructions was designed with the idea of being clear and concise but Parasep probe can be used with FE-5 Workstation unfortunately was generally found to be confusing. The Para-Pak Macro-Con Stool Concentration System was found to be particularly confounding. Both the FPC Faecal Parasite Concentrator and the FPC Jumbo Faecal Parasite Concentrators had very well presented instructions with good clear pictures demonstrating each of the steps. The Parasep range instructions are clear, concise and simple to follow in comparison to the other three products. They were designed to simplify the whole process into 4 quick and easy steps whilst all of the other products had many more steps. The main idea behind most of these commercial products centred on ease of use with an attempt to prevent operator exposure. Whilst this is a good idea, the first three products all leaked once the filter units were attached to the transport vials. Filter unit leakage was observed once filtration had commenced and also when the device was disconnected. These products should not leak due to the fact that all of the separate units that are used are specially designed for the product. Any leakage at all may compromise the operator and the concentration procedure itself. The sealing mechanism of the Parasep range prevents any leakage. Even though the Maxi-Parasep can suit both 15 and 50 ml tubes there was no leakage present. The Mini and Midi-Parasep units are supplied with integrated collection vials with a torque differential to ensure complete sealing and ease of removal following faecal concentration. In three of the products tested there was much blockage of the filter resulting in poor filtration and separation. Without the fundamental ability to efficiently filter and concentrate the faecal sample the following units did not fulfill expectations (FPC and the FPC Jumbo Faecal Parasite Concentrator and the Para-Pak Macro-Con Stool Concentration System). The Parasep range devices were seen not to become blocked at any time which was attributed to the unique filtration system. The two stage filtration matrix rejected large particles without occluding the pores. 10 Whole truth in one drop CORMAY International Bulletin

11 DIAGNOSTICS CLOSE UP Parasep, a commercial kit for faecal concentration, minimises the pieces of apparatus used and is a less hazardous procedure of comparable efficiency to the standard method. Evaluation of Parasep Faecal Parasite Concentrator M. Kettelhut, A. Moody, H. Edwards and P. L. Chiodini Hospital for Tropical Deseases, Londyn The use of a concentration method by clinical laboratories is essential to increase the sensitivity of finding ova, cysts and larvae in faecal specimens as they may be to scanty to be seen by direct microscopy. The Ridley-Allen modified formol ether sedimentation technique is the method of choice for routine use by most clinical laboratories. This procedure utilises filtration of a faecal suspension followed by solvent extraction and centrifugation. It requires several pieces of apparatus and can present potential COSHH problems. Parasep, a commercial kit for faecal concentration, developed by the company Dia- Sys Europe Limited (formaly Intersep Ltd), is an enclosed, single use disposable system which minimises the pieces of apparatus used and is a less hazardous procedure of comparable efficiency to the standard method. A comparison between the open and enclosed systems using ether or ethyl acetate with Triton-X as the solvent extraction gave a similar recovery of parasites and negligible interference in the amount of debris present in the deposit. There was, however, a significant increase in the recovery of certain parasites namely Taenia species and Hymenolepis nana, and in some specimens stored in 10% formalin at 4 C containing ova of Ascaris lumbricoides and Toxocara canis, when ethyl acetate was used. INTRODUCTION The microscopic examination of faeces is essential for the recognition and identification of intestinal parasites. Direct microscopy, although useful for the observation of motile protozoan trophozoites and the examination of cellular exudate, is not recommended solely for the routine examination of faeces with suspected parasitic infections. In order to maximise the numbers of organisms detected, a concentration method is essential to increase the possibility of recovering ova, cysts and larvae which may be too scanty to detect by direct microscopy alone. The Ridley-Allen (RA) modified formol ether technique is the procedure recommended for clinical laboratories for the routine diagnosis of parasitic infections and is the technique used routinely by the Department of Clinical Parasitology, Hospital for Tropical Diseases (HTD), London. This method utilises ether or ethyl acetate as an extractor of fat and debris from faeces after filtration and leaves the parasites in a sediment at the bottom of the tube after centrifugation. The advantages of this method are that it will recover most ova, cysts and larvae and retains their morphology thus facilitating identification. The method can also be used on samples which have been preserved in formalin, sodium acetic acid formalin (SFA) and polyvinyl alcohol (PVA), but has the disadvantage of destroying trophozoite stages and distorting cellular exudate, liquid faeces do not concentrate well thus it is ne- A concentration method is essential to increase the possibility of recovering ova, cysts and larvae which may be too scanty to detect by direct microscopy alone cessary in these cases to examine the stool by direct microscopy. This study compares the conventional open method for modified Ridley-Allen concentration technique used at the Hospital for Tropical Diseases; with Parasep, a commercial kit developed by DiaSys Europe Limited (formaly Intersep Ltd), Unit 5, The Sapphire Centre, Fishponds Road, Wokingham, Berkshire, RG41 2QL, England, which is an enclosed, single use, disposable system for the modified RA sedimentation technique. MATERIALS AND METHODS The Parasep Faecal parasite concentrator employs the principal of the Ridley-Allen formol-ether sedimentation technique in an enclosed system. It consists of a mixing chamber in which the faeces is mixed with the 10% formalin. The ether or ethyl acetate is added (1 drop of Triton-X is added to the mixture when ethyl acetate is used as it helps to break up the faecal matter) and Parasep is immediately sealed by screwing the filter/thimble sedimentation cone onto the mixing chamber. The seal is an air/liquid seal which prevents the release of biohazardous material. There is also a safety lock to ensure that the mixing chamber and filter thimble are removed together for safe disposal. Nr 2 (22), summer 2011 Whole truth in one drop 11

12 DIAGNOSTICS CLOSE UP Patented Filter Thimble (High Density Polyethylene) A two stage filtration matrix. Large particles are rejected without occluding the 425 μm pores. Recovery rate with Parasep is comparable to traditional sieve method, ie: Ridley-Allen. Mesh detail Mixing Chamber (High Clarity Polypropylene) Derbis Trap Rejected particles are trapped to prevent extrusion into the Sedimentation Cone during centrifugation. Integral Spoon Air/Liquid Seal and Safety Lock The seal prevents the release of biohazardous material. The lock ensures the Mixing Chamber and Filter Thimble are removed together for safe disposal. Sedimentation cone (High Clarity Polypropylene) Sediment forms in the base of the cone allowing examination for the presence of helminth eggs or larvae and protozoal cysts or oocysts. Centrifuge Compatibility Designed to fit all 15 ml centrifuge buckets. The standard Ridley-Allen formol-ether concentration use as by HTD follows the method as described as in Reference 1. The mixture is vortexed and Parasep is then inverted to allow the mixture to be filtered through the filter thimble. The filter thimble is made from high density polyethylene and has a 2 stage filtration matrix which means that the large particles are rejected without occluding the 425 μm pores. There is also a debris trap so that rejected particles are trapped to prevent extrusion into the sedimentation cone. (see diagram below). Parasep is then centrifuged at 3000 rpm for 1 minute. The mixing chamber and filter thimble are unscrewed and discarded. Like the conventional Ridley-Allen sedimentation method, there is a ether/ethyl acetate layer, fatty plug, formalin and sediment, The fatty plug is loosened and the supernatant discarded. The deposit is examined for ova, cysts and larvae. 100 faecal samples containing previously diagnosed ova, cysts or larvae were examined by the Ridley Allen sedimentation technique as used by the Hospital for Tropical Diseases and by the Parasep. Ether and ethyl acetate with 1 drop of Triton X, were compared in parallel as the lipid extracting agent on both techniques. The faecal samples were divided into the following categories: faecal samples containing ova, 21 of which contained only 1 species of helminth and 5 contained 2 or 3 species of helminths faecal samples containing protozoan cysts or oocysts; 15 of which contained Group 1: Specimens containing ova and larvae 1a. 21 specimens containing ova and larvae of 1 species of helminth Average number of ova per deposit Open Ridley-Allen concentration method Enclosed Parasep faecal concentrator Helminth species Ether Ethyl acetate Ether Ethyl acetate Ascaris lumbricoides x2 Hookworm species x4 Trichuris trichiura x3 Rhabditiform larvae of Strongyloides stercoralis x Filariform larvae of Strongyloides stercoralis x Toxocara canis x1 Trichostrongylus species x1 Taenia species x2 Hymenolepis nana x b. 5 mixed helminth infections (Table 2) Number of ova per deposit Open Ridley-Allen concentration method Enclosed Parasep faecal concentrator Helminth species Ether Ethyl acetate Ether Ethyl acetate Ascaris lumbricoides and Trichiuris trichiura Hookworm species and Schistosoma mansoni Ascaris lumbricoides, Trichuris trichiura and Hookworm species Ascaris lumbricoides, Trichuris trichiura and Hymenolepis nana Ascaris lumbricoides, Hookworm species and Schistosoma mansoni CORMAY International Bulletin

13 DIAGNOSTICS CLOSE UP only one species of protozoa and 9 contained 2 or more protozoa containing no ova, cysts or larvae. Some faecal samples were preserved in formalin and stored in 1ml aliquots at 4 C. The remainder of the specimens were fresh and unpreserved and examined directly. RESULTS 100 stool specimens were examined in parallel by both Ridley-Allen open method and enclosed Parasep techniques using ether and ethyl acetate. The number of ova and larvae were counted per total deposit and the number of cysts were counted per field using a x40 objective. Where there was more than 1 specimen containing the The Prasep technique has the advantage of being a disposable enclosed system thus minimising the number of pieces of apparatus used. This speeds up the whole procedure, makes it easier to handle and more user friendly same parasite, the average number of parasites present was calculated. A comparable recovery of parasites was noted in both methods. However, a significant increase was noted in the recovery of Cyst enumeration code As it is impractical to count the number of cysts/oocysts per deposit, the following code was used: Group 2: Specimens containing cysts 2a. 17 specimens containing cysts or oocysts of 1 protozoa species (Table 3) Average number of cysts/oocysts per field (x40 objective) Open Ridley-Allen Concentration Method Enclosed Parasep faecal concentrator Protozoa species Ether Ethyl Acetate Ether Ethyl Acetate Endolimax nana x2 Entamoeba histolytica x1 Entamoeba coli x3 Chilomastix mesnili x1 Giardia lamblia x3 Cyclospora cayetanensis x3 Isospora belli x b. Mixed protozoa infections (Table 4) Number of cysts/oo cysts per field (x40 objective) Open Ridley-Allen Concentration Method Enclosed Parasep faecal concentrator Protozoa species Ether Ethyl Acetate Ether Ethyl Acetate Entamoeba coli Endolimax nana Entamoeba coli Entamoeba histolytica Entamoeba histolytica lodamoeba bütschlii Cyclospora cayetanensis Endolimax nana Entamoeba histolytica Endolimax nana Entamoeba hartmanni Chilomastix mesnili Endolimax nana Entamoeba histolytica Entamoeba coli Entamoeba histolytica lodamoeba bütschlii Entamoeba Hartmanni Entamoeba coli >10 cysts per field (x 40 objective) 5 10 cysts per field (x 40 objective) 1 5 cysts per field (x 40 objective) 1 cysts per 2 10 fields (x 40 objective) - - < 1 cyst per 10 fields ova of Taenia species, Hymenolepis nana, Ascaris lumbricoides and Toxocara canis when ethyl acetate with Triton-X as a lipid extraction agent was used compared to ether. The ova of Tania species and Hymenolepis nana are light and on examination, it was noted that they had become trapped in the fatty flug and were discarded. The ova of Toxocara canis and Ascaris lumbricoides, in particular those in table 2, had been stored in formalin for long periods and may have altered their density. A comparable recovery of parasites was noted using both methods. However, there was considerably more deposit using ethyl acetate making the cysts more difficult to see without dilution. GROUP 3: 50 NEGATIVE FAECES 50 faeces were shown to contain no ova, cvsts or larvae by the modified Ridley Allen open technique and Parasep, using both ether and ethyl acetate. DISCUSSION Although a sedimentation concentration technique maximises the recovery of most ova, cysts and lanrvae, the deposit may contain some debris which, when present in excess, could mask the presence of small parasites, especially cysts. A notable difference Nr 2 (22), summer 2011 Whole truth in one drop 13

14 DIAGNOSTICS CLOSE UP was observed, however, between the final deposits when using ether or ethyl acetate, the latter resulting in much thicker deposit resulting in potential obscuring of cysts. The rapidity and safety by which the samples can be processed are factors to take into consideration when deciding on an appropriate concentration technique. The standard open Ridley-Allen sedimentation techique uses several pieces of apparatus (ie. centrifuge tubes, boiling tubes, brass seive and a collection receptacle) which can be rather labour intensive to use and clean. The Prasep technique has the advantage of being a disposable enclosed system thus minimising the number of pieces of apparatus used. This speeds up the whole procedure, makes it easier to handle and more user friendly. The use of 10% formalin as a fixative, reduces the risk of infection from bacteria and viruses. However, exposure to formalin is an irritant and ether is flammable (ethyl acetate being less flammable option), thus it is necessary to carry out this procedure in a spark proof extraction cabinet. Parasep, however, is a totally enclosed process and has an air/liquid seal and safety lock, the seal preventing the release of hazardous material and the lock ensures that the mixing chamber and filter thimble are removed together for safe disposal after centrifugation. It is also a single use device which is discarded after use so minimising the risk of sample contamination. Cost is an important consideration in deciding which concentration technique to employ. In the standard Ridley-Allen sedimentation technique, the initial purchase of brass filter, polycarbonate centrifuge tubes and boiling tubes is expensive but can be cleaned and recycled. Parasep is a single use, disposable device which can appear more expensive but can be equated with the time spent and cost of cleaning the pieces of equipment. On balance, there does not appear to be a cost advantage by either method. In summary, the recovery of parasites by the open method for the Ridley-Allen sedimentation technique and Parasep, a commercial faecal parasite concentrator, is comparable. The former procedure is lower cost but it is labour intensive and has inherent health and safety hazards. Parasep, however, is an enclosed system and a single use, disposable device thus making it less hazardous and more user friendly. We consider that the Parasep used with ethyl acetate offers a safer, more user friendly approach to faecal concentration using the Ridley-Allen formol ether sedimentation method. Parasep is a single use, disposable device which can appear more expensive but can be equated with the time spent and cost of cleaning the pieces of equipment 14 Whole truth in one drop CORMAY International Bulletin

15 DIAGNOSTICS CLOSE UP Cost saving using the Parasep faecal parasite concentrator versus the sieve method and other disposable sieve devices Methods compared are the Formal-ether Sieve method as described by HTD London and Parasep method Does note include time for microscopy or step to step transfer. Sieve Parasep Time (mins) Time (mins) 1. Take 1 g stool and add to tube. (0,15) 1. Take 1 g stool and add to tube. (0,15) 2. Decant 2 ml Formalin into tube. (0,25) 3. Seal tube and emulsify faeces by vortexing or shaking. (0,30) 4. Filter through sieve. (0,20) 5. Wash filter and discard residue. (1,30) 6. Transfer to boiling tube, add 3 ml ether and mix by vortex. (0,45) 7. Centrifuge at 3000 rpm for. (1,00) 8. Loosen plug and discard supernatant. (0,25) 9. Transfer sample to slide. (0,05) 2. Seal tube and emulsify faeces for. (0,30) 3. Insert Parasep filter. (0,20) 4. Centrifuge at 1000 g for. (1,00) 5. Loosen plug and discard supernatant. (0,25) 6. Transfer sample to slide. (0,05) 7. Seal and discard Parasep. 10. Wash all tubes. (1,00) Total: 6,15 Total: 2,35 Nr 2 (22), summer 2011 Whole truth in one drop 15

16 STAY HEALTHY Unwanted souvenirs Entamoeba Coli We travel to other continents and exotic countries. However, the escape from European modern-age diseases does not mean that our health is completely safe. What are the risks of travelling abroad and how can we protect ourselves from tropical diseases? Katarzyna Woźniak The latest developments connected with E. Coli contamination of food products showed that food poisoning spreads with an alarming speed. Bacteria are very resistant to antibiotics and they surprise us with new mechanisms of virulence. Debilitating diarrhoea, kidney involvement and, in consequence, blood cell deterioration took over 30 people s lives in Europe. PHARAOH S REVENGE The scientists emphasise that bacteria mutate at an alarming speed, but often we make it easier for the infection to enter our body. Lack of basic hygiene, especially on holiday, may lead to an especially bothersome disease travellers diarrhoea, also known as pharaoh s revenge. Holiday makers are at the greatest risk, especially in the intertropical zone. As many as 75% of cases are caused by microbes which are ingested. Poisoning is caused by the previously mentioned bacteria such as E. Coli, as well as Shigella, Canpylobacter jejuni, E. histolytica and Salmonella. Diarrhoea can also be caused by viruses or protozoa. The patient suffers from dizziness, vomiting and high temperature. We are protected from viruses by vaccinations but it is down to us to protect ourselves from bacteria, by strictly following personal hygiene rules. You must wash your hands, after you ve been to the toilet and before every meal, do not swim in open waters after strong winds or intensive rain. Boiled or mineral water should be used both for drinking and brushing your teeth. Diarrhoea is dangerous as it may lead to dehydration, especially in high temperature and high humidity environment. NOT ONLY POISONING Diarrhoea can be misleading. It is not always due to bacterial infection. Some cases may be caused by Entamoeba histolytica parasites which lead to a serious condition called amoebiasis. The biggest risk occurs mainly in the 16 Whole truth in one drop CORMAY International Bulletin

17 tropical and subtropical sphere (food and water infected with amoeba cysts). People can become infected by eating contaminated food or via contact with an infected person s faeces. Once the amoeba cysts enter the stomach, they are not be dissolved by stomach acid and proceed into the small intestine and colon. The illness can then develop gradually: the amoeba can nest on the colonic wall and activate when the immune system weakens. Ulcers, anaemia, amoebic liver abscesses can develop even a few months after the holiday. Cholera is usually associated with a XIX disease, but the type caused by Vibrio cholerae is still very dangerous in Africa. Vomiting and diarrhoea are the most common symptoms, and they lead to rapid dehydration. It is, therefore, vital to avoid raw food and contaminated water. MOSQUITO BITES According to World Health Organisation, everyday around 3,000 children in Africa die due to malaria, and every year 3,000,000 people die for the same reason world wide. Malaria is caused by Plasmodium which enters the body with the bite of a female mosquito. The first symptoms may be visible after as long as 14 days, therefore temperature and shivers are often mistaken for a cold. High temperature occurs on average every 4 days, but malaria can lead to anaemia, lung oedema and even kidney function problems. There is no vaccination for malaria and the diagnosis is based only on the test for the presence of parasites in the red blood cells. But not everyone is as attractive to mosquitoes due to the chemical composition of sweat, though it is still recommended to cover up as much of the body as possible, especially during evenings and nights, and near water. Mosquito repellents and screens are also effective. Malaria is not the only illness spread by insects. Leishmaniasis, caused by Leishmania donovani parasites, is even more dangerous, as it causes facial ulcers which are secondarily infected by bacteria. It can lead to a permanent disfiguration of the face by the ulcer scars. STAY HEALTHY FEVER IN THE TROPICS There is no real cure for hemorrhagic fever, but there are vaccinations. Yellow fever, dengue fever and Ebola are the most dangerous. They are caused by viruses and are accompanied by high temperature and gastrointestinal bleeding. Yellow fever occurs in South America and Africa. The symptoms such as loss of appetite, hematemesis and yellow colouring of skin due to liver damage can cause permanent damage to other organs. Dengue fever is spread by female Aedes mosquitoes. The symptoms are bone and joint pain and mottled skin. Vaccinations are available against yellow fever and dengue fever, but in the case of Ebola only symptomatic treatment can be implemented. The infection spreads through droplet transmission and through contact with bodily fluids of an infected person. TRIPS In order to stay healthy it is recommended to gather as much information as possible about the places you are going to visit. If vaccinations are obligatory in a particular country, you need to vaccinate earlier otherwise you may have problems getting into that country. The travel agent should inform you of any possible risks. If you are unsure of what medicines you should take with you, please contact your doctor. Personal hygiene and responsible approach are the key factors that will keep you safe. Malaria hotspots around the world This map serves as reference only and is not the source of detailed information regarding malaria-endemic areas. WHO All rights reserved Areas where the risk of malaria is high Areas where the infection rate has been decreased Nr 2 (22), summer 2011 Whole truth in one drop 17

18 KNOWING MORE A Concise Atlas Diagnostic advice: * E. histolytica ( μm) E. coli ( μm) E. histolytica ( μm) E. coli ( μm) E. histolytica/dispar ( μm) E hartmanni ( 7,5 9 μm) E. coli ( μm) Chilomastix mesnili ( 4,5 6 μm) In E. histolytica nucleus, the cariosome may be located eccentrically, trophozoite may be mistaken for an E. coli trophozoite. In young E. histolytica cysts, glycogen agglomerates similar to those present in lodamoeba cysts can be found. The most common mistake in the Entamoeba coli diagnosis occurs when cariosome is centrally located in the nucleus and peripheral chromatin in the form of similarly sized lumps is evenly distributed on the inside of the nuclear membrane, line in E. histolytica. E. hartmanni referred to as a small variation of Entamoeba coli. Unlike E. histolytica, its trophozoites do not contain erythrocytes. Chilomastix mesnili is sometimes mistaken for E. histolytica or E. hartmanni. The differentiating factor is a cytosome in Ch. mesnili. Diagnostic advice: E. nana ( 6 12 μm) Endolimax nana ( 7 9 μm) Iodamoeba butschlii Giardia lamblia ( μm) In E. nana trophozoite, the cariosome is shaped irregularly and lacks achromatic lumps present in Iodamoeba. Cyst differentiation is easier. A mature E. nana cyst has four nuclei while Iodamoeba has one. E. nana cyst lacks iodophilic vacuole which is typical for Iodamoeba. Gardia lamblia in formed stool there are cysts and in diarrhoea stool there are trophozoites. In fresh material the presence of G. lamblia is characterised by their oval shape and numerous light refracting cellular inclusions. When stained by Lugol s iodine, the cysts sometimes show basal bodies and nuclei. Stained preparation shows a light areola close to the envelope, which gives an impression that it is separate from the cytoplasm. Giardia lamblia ( 6 10 μm) Giardia laublie ( μm) Giardia lamblia ( μm) Cyclospora cayetanensis ( 8 10 μm) Diagnostic advice: Trichuris trichiura Trichuris trichiura Trichuris trichiura Ascaris lumbricoides Detection of Trichuris trichiura (human whipworm) ova usually does not cause diagnostic problems. They can sometimes be mistaken for a Trichuris vulpis (dog whipworm), but their ova are larger and more convex. Diagnostic problems may be caused by untypical structure of Ascaris lumbricoides (human roundworm) ova misshaped, with indistinct protein envelope, with missing protein envelope or unfertilised ova with no protein envelope. Ascaris Ascaris lumbricoide Ascaris unfertile 18 Whole truth in one drop CORMAY International Bulletin

19 KNOWING MORE of Parasites Toxocara cati Toxocara canis Taenia solium ( μm) Taenia saginata ( μm) Diagnostic advice: Toxocara canis (dog roundworm) ova are similar to Ascaris ova, however they are larger and less oval. T. cati ova are round, with small indentations on the surface. T. cati ova are not present in human stool. Taenia saginata (beef tapeworm) ova are similar to T. salium (pork tapeworm) and Echinococcus spp (dog tapeworm). Taenia spp. Fasciola hepatica Fasciola hepatica Fasciola hepatica Diagnostic advice: Diphyllobothrium latum Enterobius vermicularis Strongyloides stercoralis Hymenolepis nana Diphyllobathrium latum (broad tapeworm) ova are oval with indistinct lid on one end and a small nodule on the other. Enterobious vermicularis (pinworms) ova are sometimes mistaken for pollen. Strongyloides stercoralis (threadworm) rhabditiform larvae are isolated from fresh stool samples and filariform larvae are isolated from hard stool and stool stored in warm and suitably humid conditions. Hymenolepis nana ova with filament threads located between oncosphere and the envelope may be mistaken with H. diminuta ova, which are larger and have no filaments. Diagnosis of Schistosoma mansoni ova is usually easy. Schistosoma haematobium ova are similar in size to S. mansoni. Schistosoma mansoni ( μm) Schistosoma haematobium ( μm) Diagnostic advice: Ancylostoma duodenale Ancylostoma duodenale Isospora belli ( μm) Blastocystis hominis Ancylostoma duodenale (hookworm) ova a few hours after being excreted. Cryptosporidium oocysts are different size to Cyclospora oocysts. The detection rate increases after sample concentration. Cryptosporidium * Diagnostic advice has been prepared on the basis of Prof Alicja Buczek s Atlas pasożytów człowieka Nr 2 (22), summer 2011 Whole truth in one drop 19

20 FE-5 Workstati on Reduces biohazard Increases Reproducibility Standardisation Reduces analysis time Removes costs Slides Cover slips Pipettes Better morphology Allows enumeration

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