A LABORATORY GUIDE TO PARASITOLOGY CTHE FIFTH EDITIONI ZHENG XIAOYING WU ZHONGDAO Department of parasitology Zhongshan Medical collage Sun Yat-Sen University July, 2002
A LABORATORY GUIDE TO PARASITOLOGY (THE FIFTH EDITION) ZHENG XIAOYING WU ZHONGDAO Department of Parasitology Zhongshan Medical collage , Sun Yat-Sen University July, 2002
CONTENTS Care of the microscope Clonorchis sinensis Paragonimus flukes Fasciolopsis buski Schistosoma japonicum Tar Ascaris lumbricoides Trichuris trichiura Ancylostoma duodenal Necator americanus Enterobius vermicularis Wuchereria bancrofti brugia malayi Trichinella spirali Examination of alimentary helminths Entamoeba histolytica. E coli other Amoebae Giardia lam blia Trichomonas vaginalis Leishmania donovani Plasmodia Opportunistic pathogenic protozoan Mosquitoes y Sandfly, fleas, lice other blood sucking insects
1 CONTENTS Care of the microscope…………………………………….…………………1 Clonorchis sinensis…………………………………….……………….…….2 Paragonimus flukes…………………………………….……………….……5 Fasciolopsis buski…………………………………….……………….….…..7 Schistosoma japonicum…………………………………….………………...8 Tapeworm…………………………………….……………….………….…14 Ascaris lumbricoides & Trichuris trichiura ………………………………..17 Ancylostoma duodenal & Necator americanus……………………………..20 Enterobius vermicularis……………………………………………….…….22 Wuchereria bancrofti & Brugia malayi……………………………………..23 Trichinella spiralis…………………………………….……………….……25 Examination of alimentary helminths…………………………………….…26 Entamoeba histolytica, E.coli & other Amoebae…………………………...27 Giardia lamblia…………………………………….……………….……….30 Trichomonas vaginalis…………………………………….………………...31 Leishmania donovani…………………………………….……………….…32 Plasmodia…………………………………….…………………….…….…35 Opportunistic pathogenic protozoan………………………………….……..40 Mosquitoes…………………………………….……………….…….……...43 Fly…………………………………….…………………….…….…….…...45 Sandfly, fleas, lice & other blood sucking insects…………………………..46
Care of the microscope Dos 1. Do take special care to protect the microscope from dust in hot dry periods 2. Do take special care to protect the microscope lenses and prisms from fungal growth in hot humid periods 3. Do clean the immersion oil from the immersion objective each time, use lens tissue dampened with ethanol 4. Do clean the oculars with lens tissue 5. Do use the microscope retaining screw fitted at the base of the microscope box to prevent damage to the instrument while in transit Dont 1. Don't use the tissue used for the oil immersion objective to clean the oculars 2. Don't dismantle or try to clean parts of the microscope that are difficult to reach 3. Don't leave the lens parts empty; use the appropriate cover to cover the empty port 4. Don't exchange lenses from microscopes of different manufacture
2 Care of the microscope Do's 1. Do take special care to protect the microscope from dust in hot dry periods. 2. Do take special care to protect the microscope lenses and prisms from fungal growth in hot humid periods. 3. Do clean the immersion oil from the immersion objective each time; use lens tissue dampened with ethanol. 4. Do clean the oculars with lens tissue. 5. Do use the microscope retaining screw fitted at the base of the microscope box to prevent damage to the instrument while in transit. Don't 1. Don't use the tissue used for the oil immersion objective to clean the oculars. 2. Don't dismantle or try to clean parts of the microscope that are difficult to reach. 3. Don't leave the lens parts empty; use the appropriate cover to cover the empty port. 4. Don't exchange lenses from microscopes of different manufacture
CLONORCHIS SINENSIA Objectives and requirements 1. To study the morphology and the life history of C sinensis as an example of general feature of trematodes 2. To study the infection route, infection mode, parasitized site and the pathogenesis of C. sinensis infection 3. To study the characteristics of ovum of C. sinensis and the pathogenic diagnosis of C. sinensis infection 4. To learn the basic skill of drawing eggs Observation and experiment 1. Adult worm (1)Observe the preserved specimen(Demonstration) Note the size, color, shape of the organism. Spindle-shaped, transparent (2)Observe the general feature of C. sinensis in stained specimen( Manipulation First with naked eye, then study under microscope, note and identify following A. Adherent organs: oral and ventral suckers, compare their size. B. Digestive system: mouth, pharynx, esophagus and ceca C. Excretory system: excretory bladder and pore D. Reproductive sys tem: hermaphroditism Male: observe the number, size, shape and position of testes Female: observe ovary, seminal receptacle, vitellarium, Mehlis' gland and uterus What is meant by hermaphroditism? 2. Larva and intermediate hosts (Demonstration) (1) See live sporocyst, redia and cercaria(or stained sporocyst to replace live one if (2)See the stained cercaria Note the eye-spots and characteristics of tail lipoidal shape, 138 u mX115 u m, with two suckers(oral sucker an entral sucker) and excretory bladder containing black granules (4)See the first intermediate hosts(Parafossarulus, Alocinma, Bithynia snails), and second intermediate hosts, fresh water fish and crayfish. What are their names? 3. Ovum(Manipulation) (1)Study the ova. Note the shape, color, shell, operculum resting on a rim which takes
3 CLONORCHIS SINENSIS Objectives and Requirements 1. To study the morphology and the life history of C. sinensis as an example of general feature of trematodes. 2. To study the infection route, infection mode, parasitized site and the pathogenesis of C. sinensis infection. 3. To study the characteristics of ovum of C. sinensis and the pathogenic diagnosis of C. sinensis infection. 4. To learn the basic skill of drawing eggs. Observation and Experiment 1. Adult worm (1) Observe the preserved specimen (Demonstration). Note the size, color, shape of the organism. Spindle-shaped, transparent. (2) Observe the general feature of C. sinensis in stained specimen (Manipulation). First with naked eye, then study under microscope, note and identify following structures. A. Adherent organs: oral and ventral suckers, compare their size. B. Digestive system: mouth, pharynx, esophagus and ceca. C. Excretory system: excretory bladder and pore. D. Reproductive system: hermaphroditism. Male: observe the number, size, shape and position of testes. Female: observe ovary, seminal receptacle, vitellarium, Mehlis’ gland and uterus. What is meant by hermaphroditism? 2. Larva and intermediate hosts (Demonstration) (1) See live sporocyst, redia and cercaria (or stained sporocyst to replace live one if necessary). (2) See the stained cercaria. Note the eye-spots and characteristics of tail. (3) Live metacercaria. Note its ellipsoidal shape, 138μm×115μm, with two suckers (oral sucker and ventral sucker) and excretory bladder containing black granules. (4) See the first intermediate hosts (Parafossarulus, Alocinma, Bithynia snails), and second intermediate hosts, fresh water fish and crayfish. What are their names? 3. Ovum (Manipulation) (1) Study the ova. Note the shape, color, shell, operculum resting on a rim which takes
the shape of distinct shoulders, a small protuberance at abopercular end,an asymmetrical miracidium inside the egg. One of the smallest helminth ova yellow-brown, thicker eggshell Material examined: feces and drawn liquid from duodenum Examining method: see examination of alimentary helminths (2)See the scanning electron-microscopic(SEM) photograph of ovum(Demonstration) 4. Pathology (1)See specimen, note the parasites in billiary passages(Demonstration) (2)See adult worms inhabiting in billiary passages of a reservoir host(Demonstration) Exercise I. Draw egg of C. sinensis in detail 2. Label in full an adult worm of C. sinensis 3. Draw a life cycle draft of C. sinensis, including final host, intermediate host; adult parasitized site, main injured organs; excretory route of ovum, pathogenic diagnostic methods the first and se intermediate hosts and in which how the larva development; the infective infection mode and infection route Thinking 1. Why one can obtain parasitic disease after eaten uncooked fish? 2. Record the results of pressing fish flesh and the dissection of the experimental animal Reference I. The fertilization process and egg formation In male, sperm produced in a pair of testes are collected in the seminal vesicle through the vas efferens and vas deferens and are ejected through the ejaculatory duct, around which there is a prostate gland covered by a cirrus sac. In female, a short oviduct comes out of the ovary and joins with the seminal receptacle and Laurers canal opening on the dorsal side, and further proceeds to the ootype with surrounding Mehlis'gland. Then, it becomes the uterus, which extends with many contortions to the common genital pore. Small ducts from the vitelline glands, gradually gather to become the right and left vitelline ducts and then one common vitelline duct, which opens at front of the ootype An egg cell from ovary and a sperm from testis are united, additional yolk cells provided by the vitelline gland join, and eggshell formation is started in the ootype by the effect of Mehlis' gland and completed during passage through the uterus. In the egg one miracidium has already been formed 2. Examine for encysted metacercaria in fish flesh
4 the shape of distinct shoulders, a small protuberance at abopercular end, an asymmetrical miracidium inside the egg. One of the smallest helminth ova, yellow-brown, thicker eggshell. Material examined: feces and drawn liquid from duodenum. Examining method: see examination of alimentary helminths. (2) See the scanning electron-microscopic (SEM) photograph of ovum (Demonstration). 4. Pathology (1) See specimen, note the parasites in billiary passages (Demonstration). (2) See adult worms inhabiting in billiary passages of a reservoir host (Demonstration). Exercise 1. Draw egg of C. sinensis in detail. 2. Label in full an adult worm of C. sinensis. 3. Draw a life cycle draft of C. sinensis, including final host, intermediate host; adult parasitized site, main injured organs; excretory route of ovum, pathogenic diagnostic methods; the first and second intermediate hosts and in which how the larva development; the infective stage, infection mode and infection route. Thinking 1. Why one can obtain parasitic disease after eaten uncooked fish? 2. Record the results of pressing fish flesh and the dissection of the experimental animal. Reference 1. The fertilization process and egg formation In male, sperm produced in a pair of testes are collected in the seminal vesicle through the vas efferens and vas deferens and are ejected through the ejaculatory duct, around which there is a prostate gland covered by a cirrus sac. In female, a short oviduct comes out of the ovary and joins with the seminal receptacle and Laurer’s canal opening on the dorsal side, and further proceeds to the ootype with surrounding Mehlis’ gland. Then, it becomes the uterus, which extends with many contortions to the common genital pore. Small ducts from the vitelline glands, gradually gather to become the right and left vitelline ducts and then one common vitelline duct, which opens at front of the ootype. An egg cell from ovary and a sperm from testis are united, additional yolk cells provided by the vitelline gland join, and eggshell formation is started in the ootype by the effect of Mehlis’ gland and completed during passage through the uterus. In the egg one miracidium has already been formed. 2. Examine for encysted metacercaria in fish flesh