Preface xix About the Author xxiii Section 1 Philosophy and Approach to Diagnostic Parasitology 1 Neglected Tropical Diseases 2 Why Perform Diagnostic Parasitology Testing? 3 Travel 3 Population Movements 3 Control Issues 4 Climate Change 4 Epidemiologic Considerations 4 Compromised Patients; Potential Sex Bias Regarding Infection Susceptibility; Aging 4 Approach to Therapy 5 Who Should Perform Diagnostic Parasitology Testing? 5 Laboratory Personnel 5 Nonlaboratory Personnel 5 Where Should Diagnostic Parasitology Testing Be Performed? 6 Inpatient Setting 6 Outpatient or Referral Setting 6 Decentralized Testing 6 Physician Office Laboratories 6 Over-the-Counter (Home Care) Testing 7 Field Sites 7 What Factors Should Precipitate Testing? 7 Travel and Residence History 7 Immune Status of the Patient 7 Clinical Symptoms 7 Documented Previous Infection 7 Contact with Infected Individuals 8 Potential Outbreak Testing 8 Occupational Testing 8 Therapeutic Failure 8 What Testing Should Be Performed? 8 Routine Tests 8 Special Testing and Reference Laboratories 9 Specialized Referral Test Options--DPDx and Other Sites 9 Other (Nonmicrobiological) Testing 10 What Factors Should Be Considered in Development of Test Menus? 10 Physical Plant 10 Client Base 10 Customer Requirements and Perceived Levels of Service 10 Personnel Availability and Level of Expertise 10 Equipment 10 Budget 11 Risk Management Issues Associated with Stat Testing 11 Primary Amebic Meningoencephalitis 11 Granulomatous Amebic Encephalitis and Amebic Keratitis 14 Request for Blood Films 15 Automated Instrumentation 15 Patient Information 16 Conventional Microscopy 16 Table 1.1 Common features of the neglected tropical diseases 18 Section 2 Parasite Classification and Relevant Body Sites 19 Protozoa (Intestinal) 20 Amebae, Stramenopiles 20 Flagellates 22 Ciliates 22 Apicomplexa (Including Coccidia) 23 Microsporidia (Now Classified with the Fungi) 23 Protozoa (Other Body Sites) 24 Amebae 24 Flagellates 24 Apicomplexa (Including Coccidia) 24 Microsporidia (Now Classified with the Fungi) 25 Protozoa (Blood and Tissue) 26 Apicomplexa (Including Sporozoa) 26 Flagellates 26 Nematodes (Intestinal) 29 Nematodes (Tissue) 29 Nematodes (Blood and Tissue) 30 Cestodes (Intestinal) 31 Cestodes (Tissue) 32 Trematodes (Intestinal) 33 Trematodes (Liver and Lungs) 33 Trematodes (Blood) 35 Pentastomids 35 Acanthocephala 36 Table 2.1 Classification of human parasites 37 Table 2.2 Cosmopolitan distribution of common parasitic infections 39 Table 2.3 Body sites and possible parasites recovered 40 Section 3 Collection Options 43 Safety 44 Collection of Fresh Stool Specimens 44 Collection Method 44 Number of Specimens To Be Collected 45 Collection Times 47 Posttherapy Collection 47 Specimen Type, Stability, and Need for Preservation 47 Preservation of Stool Specimens 48 Overview of Preservatives 48 Formalin 49 Sodium Acetate-Acetic Acid-Formalin (SAF) 50 Schaudinn''s Fluid 51 Schaudinn''s Fluid containing PVA (Mercury Base) 52 Schaudinn''s Fluid containing PVA (Copper Base, Zinc Base) 52 Single-Vial Collection Systems (Other than SAF) 53 Universal Fixative (Total-Fix) 54 Quality Control for Preservatives 54 Procedure Notes for Use of Preservatives (Stool Fixative Collection Vials) 55 Procedure Limitations for Use of Preservatives (Stool Fixative Collection Vials) 55 Collection of Blood 56 Collection and Processing 56 Stat Test Requests and Risk Management Issues 57 Collection of Specimens from Other Body Sites 57 Table 3.1 Fecal specimens for parasites: options for collection and processing 58 Table 3.2 Approaches to stool parasitology: test ordering 61 Table 3.3 Preservatives and procedures commonly used in diagnostic parasitology (stool specimens) 62 Table 3.

4 Advantages of thin and thick blood films 64 Table 3.5 Advantages and disadvantages of buffy coat films 64 Table 3.6 Potential problems of using EDTA anticoagulant for the preparation of thin and thick blood films 65 Table 3.7 Body sites and possible parasites recovered (trophozoites, cysts, oocysts, spores, adults, larvae, eggs, amastigotes, and trypomastigotes) 66 Section 4 Specimen Test Options: Routine Diagnostic Methods and Body Sites 69 Ova and Parasite Examination of Stool Specimens 70 Other Diagnostic Methods for Stool Specimens 72 Culture of Larval-Stage Nematodes 72 Estimation of Worm Burdens through Egg Counts 73 Hatching Test for Schistosome Eggs 73 Screening Stool Samples for Recovery of a Tapeworm Scolex 74 Testing of Other Intestinal Tract Specimens 74 Examination for Pinworm 74 Sigmoidoscopy Material 75 Duodenal Drainage Material 76 Duodenal Capsule Technique (Entero-Test) 76 Urogenital Tract Specimens 77 Sputum 77 Aspirates 78 Biopsy Specimens 78 Blood 79 Thin Blood Films 79 Thick Blood Films 80 Blood Staining Methods 80 Buffy Coat Films 80 QBC Microhematocrit Centrifugation Method 80 Knott Concentration 81 Membrane Filtration Technique 81 Culture Methods 81 Animal Inoculation and Xenodiagnosis 82 Antibody and Antigen Detection 82 Antibody Detection 82 Antigen Detection, Nucleic Acid-Based Tests, and Molecular Panels 84 Intradermal Tests 84 UV Autofluorescence 84 Table 4.1 Body sites, procedures and specimens, recommended methods and relevant parasites, and comments 86 Table 4.2 Serologic, antigen, and probe tests used in the diagnosis of parasitic infections 94 Section 5 Specific Test Procedures and Algorithms 95 Microscopy 96 CALIBRATION OF THE MICROSCOPE 96 Ova and Parasite Examination 99 DIRECT WET FECAL SMEAR 99 CONCENTRATION (Sedimentation and Flotation) 104 SEDIMENTATION CONCENTRATION (Formalin-Ethyl Acetate) 105 SEDIMENTATION CONCENTRATION USING THE UNIVERSAL FIXATIVE (Total-Fix) 109 FLOTATION CONCENTRATION (Zinc Sulfate) 110 PERMANENT STAINED SMEAR 114 Stains Used in the Permanent Stained Smear 116 TRICHROME STAIN (Wheatley''s Method) 116 IRON HEMATOXYLIN STAIN (Spencer-Monroe Method) 122 IRON HEMATOXYLIN STAIN (Tompkins-Miller Method) 127 MODIFIED IRON HEMATOXYLIN STAIN (Incorporating the Carbol Fuchsin Step) 128 CHLORAZOL BLACK E STAIN 131 Specialized Stains for Coccidia and Microsporidia 132 KINYOUN''S ACID-FAST STAIN (Cold Method) 132 MODIFIED ZIEHL-NEELSEN ACID-FAST STAIN (Hot Method) 136 CARBOL FUCHSIN NEGATIVE STAIN FOR CRYPTOSPORIDIUM (W. L. Current) 139 RAPID SAFRANIN METHOD FOR CRYPTOSPORIDIUM (D.

Baxby) 139 RAPID SAFRANIN METHOD FOR CYCLOSPORA , USING A MICROWAVE OVEN (Govinda Visvesvara) 139 AURAMINE O STAIN FOR APLICOMPLEXA (INCLUDING COCCIDIA) (Thomas Hänscheid) 140 MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Weber, Green Counterstain) 144 MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Ryan, Blue Counterstain) 146 MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Evelyn Kokoskin, Hot Method) 149 Fecal Immunoassays for Intestinal Protozoa 150 ENTAMOEBA HISTOLYTICA 150 CRYPTOSPORIDIUM SPP. 151 GIARDIA LAMBLIA 151 KITS UNDER DEVELOPMENT 151 COMMENTS ON THE PERFORMANCE OF FECAL IMMUNOASSAYS 151 ENZYME IMMUNOASSAYS (Antigen Detection, No Centrifugation Recommended) 152 FLUORESCENCE (Visual Identification of the Organisms, Centrifugation Recommended) 152 LATERAL-FLOW CARTRIDGES (Antigen Detection, No Centrifugation Recommended) 153 Larval Nematode Culture 153 HARADA-MORI FILTER PAPER STRIP CULTURE 153 BAERMANN CONCENTRATION 155 AGAR PLATE CULTURE FOR STRONGYLOIDES STERCORALIS 158 Other Methods for Gastrointestinal Tract Specimens 161 EXAMINATION FOR PINWORM (Cellulose Tape Preparations) 161 SIGMOIDOSCOPY SPECIMENS (Direct Wet Smear) 163 SIGMOIDOSCOPY SPECIMENS (Permanent Stained Smear) 166 DUODENAL ASPIRATES 168 Methods for Urogenital Tract Specimens 171 RECEIPT OF DRY SMEARS 171 DIRECT SALINE MOUNT 171 PERMANENT STAINED SMEAR 174 URINE CONCENTRATION (Centrifugation) 176 URINE CONCENTRATION (Nuclepore Membrane Filter) 179 Preparation of Blood Films 181 THIN BLOOD FILMS 182 THICK BLOOD FILMS 184 COMBINATION THICK-THIN BLOOD FILMS 186 RISK MANAGEMENT ISSUES ASSOCIATED WITH BLOOD FILMS 187 USE OF A REFERENCE LABORATORY FOR PARASITE BLOOD DIAGNOSTIC TESTING 188 BLOOD FILM REPORTING WITH ADDITIONAL REPORT COMMENTS 189 BUFFY COAT BLOOD FILMS 189 Blood Stains 190 STAIN OPTIONS 190 GIEMSA STAIN 190 Blood Concentration 194 BUFFY COAT CONCENTRATION 194 KNOTT CONCENTRATION 195 MEMBRANE FILTRATION CONCENTRATION 197 Algorithm 5.1 Procedure for processing fresh stool for the O&P examination 201 Algorithm 5.2 Procedure for processing liquid specimens for the O&P examination 202 Algori.