- Biotechnology : An Overview (John I. Bruce)
- Strengthening Biotechnology Research in Indonesia (S. Sastrapradja and D.S. Sastrapradja)
- The Current Situation of Parasitic Infections in Indonesia (Sri Oemijati)
- Biotechnology of the Filaria of Indonesia (F. Partono and B. Rukmono)
- Importance Nematode infections in Indonesia (Sri Oemijati)
- Review on Important Helminthic Diseases in Animal in Indonesia (I.G.P. Suwata)
- Review of Research in Veterinary Parasitology in the Research Institute for Veterinary Science (RIVS) since 1983 (S. Partoutomo and Suhardono)
- Filarial Antigents : Targets for Diagnosis, Protection and Pathology (R.M. Maizels et al)
- Cestodes in Man in Indonesia (Sri S. Margono)
- The Application of Biotechnology to the Study of Cetodes (John H. Cross)
- Application of Biotechnology to the Study of Filarial Parasites and their Vectors (A.C. Vickery)
- Antigenaemia as an Indicators of Filarial Endemicity (F. Partono et al)
- A Field Study Using the Polymerase Chain Reaction (PCR) to Screen for Brugia Microfilariae in Hman and Animal Blood (Janet Glover et al)
- Detection of Brugia malayi Infected Mosquitoes with Species Specific DNA Probe pBm 15, in Riau, Indonesia (L. Kurniawan et al)
- Detection of Brugia malayi and Brugia pahangi Parasites by Biotinlabeled DNA Probes (T. Supali et al)
- The Tubulin Gene of D.immitis (Nur Asikin)
- Use of Melocular Genetic Engineering in the Study of Animal Parasites and Their Vectors (Philip T. Loverde)
- Important Trematodes in Man in Indonesia (Pinardi Hadidjaja)
- Current Technological Approaches to the Study of Trematodes (W.P. Carney)
- Molecular Approach to the Study of Trematode Parasites : The Blood Fluke (Philip T. LaVerde)
- The Application of a Numerical Method for Taxonomy from Isoenzymatical Result : Coefficient of Parental Correlation Between Schistosoma Species (Sri Subekti Bendryman Soedjoko)
- The Medically Important Molluscs of Indonesia (Machfudz Djajasasmita)
- Biotechnology in the Study of Molluscs (Christopher J. Bayne)
- Importan Protozoan Parasites in Indonesia (Srisasi Gandahusada)
- Important Protozoan Diseases of Animals in Indonesia ( a review) (Soeprapto Soekardono)
- Biotechnology in the Study Protozoan Parasites (Donald E. Burgess)
- Detection of Plasmodium Falciparum in Blood Using DNA Probe, pPF 14 (Syahrial Harun et al)
- Vector of Malaria and Filariasis in Indonesia (Hoedojo)
- Multiplication of Dengue and Chikungunya Viruses in Aedes Mosquitoes (Soedarto Soekiman)
- The Sporozoite Enzyme-Linked Immunosorbent Assay : Application in Malaria Epidemiology (Michael J. Bangs)
- Seroepidemiologic Correlations in Malaria (Kevin Baird)
- Plaque Assay of Newcastle Disease Virus (B. Sardjono)
- The Murine Cell Surface Antigen PC-1 as a Marker for Plasma Cell (Alida R. Harahap; James W. Gooding)
- Anti-NP Immunoglobulin Gene : Somatic Hypermutation Does not Directly Influence its Specificity Change from Heteroclitic to Homoclitic (Amin Seobandrio W.K.)
- Tailoring Immunotoxin as Anticancer Drug (Santoso Cornain et al)
- Opprtunities in the United States for Training in Biotechnology for Parasitic Disease Investigations (W.P. Carney)
- Soil-Transmitted Helminths in Yogyakarta (Noerhayati Soeripto et al)
- Hybridoma Production Using Immunee Lymphocytes Against Brugia malayi Antigen with Myeloma Cells (Soeyoko; Noerhajati S; Abdul S. Safro)
- Malaria in Central Java (Sugeng Juwono Mardihusodo)
- Isolation of Bacillus sphaericus and Related forms Pathogenic to Culex quinquwfasciatus (J. SItumorang et al)
- Human Parasitic Infections in Bali : A Review (Putu Sutisna)
The Inceidance of parasitic Diseases inLivestock in Bali (N.A. Suratma)
Biotechnology : An Overview
John I. Bruce
Abstract :
Biotechnology as a science includes various aspects of the management and manipulation of biotechnological systems. Recent advances in immunology, molecular biology, cell culture and other associated areas provide an opportunity for scientists to move biology out of the laboratory and into the realms of society. This has many implications which mankind on a whole may not be prepared to cope whit at this time. This new capability has been referred to as ‘Biotechnology’. Biotechnology has also been defined as ‘the integrated use of biochemistry, microbiology, and chemical engineering in order to achieve the capacities of microbes and culture cells’. Genetic engineering which includes gene splicing and recombinant DNA-cloning is an example of a recent offshoot of biotechnology.
Because of the advent of biotechnology, one can now think of the prospect of engineering tomorrows vaccines. In the past, vaccine development has been laborious and in many instances an unrewarding task. After years of effort only a handful of safe, effective vaccines have emerged. In the biotechnology arena, new methodologies and strategies for immunizing humans and domestic animals against infectious diseases are providing new hope for discovering successful vaccines. While most of the effort in the past has focused on viral vaccine development, attention is now being directed towards vaccines for protection against parasitic diseases.
Currently, considerable effort is being made to develop vaccines for malaria, coccidiosis (in fowl), cholera, malaria, schistosomiasis and trypanosomiasis among others.
Strengthening Biotechnology Research in Indonesia
S. Sastrapradja and D.S. Sastrapradja
Abstract :
The wave of biotechnology promises has struck not only the developed countries but the developing countries as well. The scientific community in Indonesia is aware of the opportunities and is eager to take an active part in this particular endeavour. Meanwhile resources are required to welcoming the biotechnology era. The need of trained manpower, appropriate infrastructure and equipment, operational and maintenance costs requires serious consideration if a unit or a laboratory is expected to be functional in biotechnology. There is a good opportunity of applying biotechnology in the field of agriculture and industry considering the availability of biological resources in Indonesia. This paper outlines what have been done so far, the difficulties encountered and the efforts made to strengthening biotechnology research in Indonesia.
The Current Situation of Parasitic Infections in Indonesia
Sri Oemijati
Abstract :
Parasitic infections are highly prevalent in Indonesia, especially in rural areas, subuds and slums of big cities. Twenty two species of protozoa and 32 species of helmiths have been reported infecting man in Indonesia. Among the 16 species of intestinal protozoa, nine are constantly found in stool surveys, but only Entamoeba histolytica and Giardia lamblia are real pathogens. Among the blood and tissue protozoa, the most important are the malaria parasites. The most frequently encountered and widely distributed species are Plasmodium falciparum, and P. vivax. P. malariae is at present more difficult to find, while P. ovale has been reported only from Flores, Timor and Irian Jaya. The non human parasites so far has not been diagnosed in human. Among the 80 species of Anopheline mosquitoes in Indonesia, 16 have been reconfirmed as vectors. Among the other tissue protozoa, Trichomonas vaginalis is frequently found in the Gynaecological clinic, while Toxoplasma gondii is found only in special studies.
Among the 13 species of intestinal nematodes, five are highly prevalent namely : Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale, Trichuris trichiura and Oxyuris vermicularis, while Strongyloides stercoralis is getting more difficult to find. Filariasis is widely distributed and is still highly endemic in certain areas. Both urban and rural Wuchereria bancrofti are prevalent, but B. malayi is causing more public health problems in rural areas. Both the human and the zoonotic type are prevalent B. timori so far has been described only from the south eastern part of Indonesia. The filarial worms have different vectors and are therefore different in epidemiology and distribution. Non human filarial worms have not been reported infecting man in Indonesia. Among the 12 species of Trematodes, only Schistosomiasis japonicum is endemic in Central Sulawesi, and recently an endemic area of Fasciolopsis buski was discovered in a restricted area in South Kalimantan. Echinostoma lindoense which was highly endemic in lake Lindu area has disappeared completely. Among the 8 species of Cestodes, Taenia saginata and T. solium with its cysticercus are found more often. While endemic in a few areas only, cases been reported from many places. Infections with other Cestodes have been reported occasionally.
On the whole, parasitic infections in Indonesia have been decreasing. Some parasites which were prevalent in the order days, have dissappeared, and human infections with animal parasites have been reported occasionally because of the difficult diagnosis.
Biotechnology of the Filaria of Indonesia
F. Partono and B. Rukmono
Abstract :
More than 90 million people were currently infected with lymphatic filariasis and two-third of them lived in China, India and Indonesia. Filariasis is endemic throughout thr intire Indonesian Archipelago. More than 20 millioan people lived in endemic areas and 3-4 ,illion people were estimated to have the infection. Control measueres have reduced the prevalence of infrctions in some areas, but the desease remained a public health problem in many outer islands of Indonesia. Recent development in monoclonal atibodies and recombinant technology of DNA have made it possible to apply this new tools in the studies of filariasis, and three groups in Indonesia are currently using this technology. The studies with the Imperial College of London will be presented by Dr. Rick Maizels. Collaboration with the new England Biolabs and Smith College involve the use of a stage and species-specific monoclonal antibody against the infective larvae of Brugia malayi, a double blind comparison of conventional methods and DNA probes for the diagnosis of brugian filariasis, and phylogentic studies of the brugian parasities. The ELISA using the monoclonal antibody has been adapted for field use in Jakarta. It is simple to use, does not cross-react with the infective stage of Brugia pahangi, but does so with the infective use of the non-symphatric Brugia timori. The reagent is usefull to acurately monitor the progress of control programs in endemic areas of brugian filariasis. The oligonucleotide DNA probes for B. malayi and B. pahangi were both qualitatively comparable to the conventional methods for the diagnosis of brugian parasites in cats and man. Sequencing data of the repeated DNA sequences of various brugian parasites indicated their homologies and divergences. The anthropophilic strain of B. malayi and B. timori showed similarity in their biological characteristics and repeated DNA sequences and they are phylogenically probably closely related. One isolate of B. malayi from Tanjung Pinang showed closer homology to the repeated DNA sequences of B. pahangi than to that B. malayi. Studies on the repeated DNA sequences of different isolated of brugian parasites are essential before DNA probes can be widely used in field studies.
Importance Nematode infections in Indonesia
Sri Oemijati
Abstract :
At least 13 species of intestinal nematodes and 4 species of blood and tissue nematodes have been reported infecting man in Indonesia. Five species of intestinal nematodes are very common and highly prevalent, especially in the rural areas and slums of the big cities. Those species are Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale, Trichuris trichiura and Oxyuris vermicularis, while Strongyloides strecoralis is disappearing. The prevalence of the soil transmitted helmints differs from place to place, depending on many factors such as the type of soil, human hehaviour etc. Three species of lynph dwelling filarial worms are known to be endemic, the urban Wuchereria bancrofti is low endemic in Jakarta and a few other cities along the north coast of Java, with Culex incriminated as vector, high endemicity is found in Irian Jaya, where Anopheline mosquitoes act as vectors. Brugia malayi is widely distributed and is still highly endemic in many areas. The zoonotic type is mainly endemic in swampy areas, and has many species of Mansonia mosquitoes as vectors. B. timori so far has been found only in the south eastern part of the archipelago and has Anopheles barbirostris as vector.
Human infections with animal parasites have been diagnosed properly anly when adult stages were found either in autopsies or removed tissues. Cases of infectious with A. caninum, A. braziliense, A. ceylanicum, Trichostrongylus colubriformis, T. axei and Oesophagostomum apiostomum have been described from autopsies, while infections with Gnathostoma spiningerum have been reported from removed tissues. Infections with the larval stages such as VLM, eosinophylic meningitis, occult filariasis and other could only be suspected, since the diagnosis was extremely difficult and based on the finding and identification of the parasite. Many cases of creeping eruption which might be caused by the larval stages of A. caninum and A. braziliense and Strongyloides stercoralis were diagnosed only clinically. Also diagnosed clinically were occult filariasis which was caused by animal filarial worms, and eosinophylic meningitis caused by Angiostrongylus cantonensis.
While Toxocara canis and T. cati are prevalent in dogs and cats, Anisakis in marine fish, no cases of Visceral larva migrans and anisakiasis have been reported in man in Indonesia.
Review on Important Helminthic Diseases in Animal in Indonesia
I.G.P. Suwata
Abstract :
Helminthic diseases are widely spread throughout the world. In Indonesia, the cases in animals are primarily associated with the condition of the field, although the intensity of the infestations are also affected by various factors inside the body of the host.
In general, the tropical and humid conditions in Indonesia, optimally support the development and spreading of the parasites, so that the prevalence of the infestations are usually high except in the very dry areas.
In Indonesia, important helminthic diseases found in livestock are mostly caused by nematodes and trematodes, and there is a lack of information regarding cestode infestations, except infestation by immature stages of the worm such as cysticercosis in ruminants and swine. On the other hand, dogs and cats are usually infested by cestodes and nematodes.
Here, the negative influence of helminthic infestation on live stock is mostly shown by failure of growth, decrease of body weight and body resistance, demage of organs infested by the parasites, but it is not rare that the disease cause death of the infested animals such as haemonchiasis in sheep, ascariasis in young swine and calves, etc.
The integrated system of farming combined with periodic anthelminthic treatments were favourable in the effort of controlling the disease.
Review of Research in Veterinary Parasitology in the Research Institute for Veterinary Science (RIVS) since 1983
S. Partoutomo and Suhardono
Abstract :
Studies on gastrointestinal parasites and the epidemiology and pathogenesis of blood protozoa in ruminants have been undertaken during the last few years. An abattoir survey for a period of one year showed that most large ruminants slaughtered in the Bogor abattoir were infected with nwmatodes, about half with trematodes and few with blood parasities. Worm infections were also common in small ruminants but none were infected with blood protozoa. Some field studies on Fasciola infection in large ruminants showed that these animals were susceptible to the disease. The seasonal prevalence of fascioliasis is not yet fully determined; management systems and the local geography play an important role. Most anthelminthics were effective against worm infection in large and small ruminants. A new flukicide (triclabendazole) is highly effective against both immature and mature liver flukes. Studies on the epidemiology and pathogenesis of trypanosomiasis in cattle and buffalo have been conducted since 1982. The result indicate that T. evansi is prevalent in five regions in Java; the prevalence rates are higher in Sahiwal and Belmont Red than Ongole cattle or buffalo. ELISA values increase with increasing age of the animals. Buffaloes are more susceptible than cattle. Naganol is the drug of choice for control of trypanosomiasis in Indonesai.
Filarial Antigents : Targets for Diagnosis, Protection and Pathology
R.M. Maizels; M.E. Selkirk; G.E. Lim; W.F. Gregory; M. Yazdanbakhsh; F. Partono
Abstract :
A range of surface, secreted and somatic antigens from filarial parasites have been studies in order to analyse the response of human infected with these pathogens, and to develop reliable diagnostic and prophylactic agents. Diagnostic procedures, which are urgently required for targetting chemotherapy, are being developed by two techniques. Firstly, detection of host antibody is carried out using selected, specific parasite antigens in the form of recombinant peptides from a filarial DNA library. Secondly, mesurement of parasite by a monoclonal antibody ‘antigen-capture’ assay. In addition, a longer-term objective of our collaborative study is to isolate molecules which may stimulate the immune system to mount a protective immune respone against filarial parasites. A major focus has been a parasite surface glycoprotein known to be closely conserved between adult worms of Brugia malayi, B. timori and Wuchereria bancrofti. This antigen has been cloned from cDNA library, and its primary sequence established; in addition to being a constant feature of the adult surface, it is expressed by developing larvae and represents an attractive target for vaccine production. Finally, one of the most intriguing questions in filariasis relates to the genesis of pathological reactions. Although this is a difficult problem, we are now beginning to compare the immune response of individuals of differing clininal status to certain defined parasite antigens, in an attempt to correlate disease development with particular categories of immune response in infected patients. In this way there is hope to advance the basic understanding of filarial disease, while providing practical means for controlling filariasis at the individual and community levels.
Cestodes in Man in Indonesia
Sri S. Margono
Abstract :
Cestodes are found endemic in certain areas of Indonesia. The most common cestodes found are Taenia saginata and Taenia solium. Eggs of Taenia are found in stool samples during surveys in Irian Jaya (2-9 %), Nusa Tenggara Timur (7 %), Bali and resettlement areas of people from Bali (0,4-3,3 %). Interviews, with questions concerning expelled segments, recovered a prevalence of 9,5% in inhabitants of the island of Samosir (North Sumatera) and 2 % in the people of 6 villages in Abiansemal (Bali). Cases are also reported from Jakarta, Hymenolepis nana (0,2-1 %) and Hymenolepis diminuta (0,4 %) are rarely found in surveys. A case of hyperinfection with H. nana has been reported in Jakarta in 1968. Occasionally there are reports of infections with Dipylidium caninum, Raillietina madagascariensis, Bartiella studeri and sparganum.
Report on treatment of taeniasis in Indonesia mentioned the use of atabrine, mebendazole, bithionol and praziquantel with different results.
The Application of Biotechnology to the Study of Cetodes
John H. Cross
Abstract :
Cestodes to tapewowms are found in vertebrate hosts worldwide. There are a great many species, but few have received much attention in biotechnologic research. Those that have been studied in any detail have been those of importance to veterinary and human medicine. The application of biotechnology to the study of taeniids has been gaining momentum in recent years. Research has been done to improve the diagnosis of larval taeniid infections, especially cysticercosis. There have been improvements in serologic testing using refined and purified antigents readily available from one species to detect antibodies of another. (Taenia hydatigena antigens are used to detect Cysticercus bovis and Cysticercus granulosus) has been shown to be effective in neurocysticercosis and hydatid diseases.
Studies with monoclonal antibodies have also been found to be of interst. Anti-oncospheral monoclonal antibodies have been developed to distinguish eggs of E. granulosis from other taeniid eggs. In another study, monoclonal antibodies from ancospheres of T. saginata conferred protection against oral infection with T. saginata eggs in calves. Other investigation reported vaccines against Cysticercus fasciolaris by a T. taeniaeformis antigen expressed in Escherichia coli.
Stadies on DNA have been gaining momentum. DNA-based techniques have been used to detect inter and intraspecific variations in Echinococcus and to characterize isolates of E. granulosus. DNA probes in Southern blot analysis have been used to discriminate taeniid species.
Taenia saginata in the Far East has become an enigma. Although the parasite is morphologically T. saginata, the definitive host for the parasite is not clear. Studies have shown the pig to be a possible intermediate host with larval development similar to T. solium, but in pig liver rather than muscle. In recent comparative studies with T. saginata-like worms from Taiwan and classical T. saginata, differences have been detected in DNA hybridization patterns.
Application of Biotechnology to the Study of Filarial Parasites and their Vectors
A.C. Vickery
Abstract :
Over 200 species of filarial parasites have been described, although the life cycle and nature of their obligate intermediate arthropod vectors have been identified for only about a quarter of them. Traditional methods of studying phylogenetic relationship between closely related parasite species have utilized morphologic, biochemical and biologic characteristics, usually of the microfilarial stage. Identification of component vectors from among complexes of sibling species, has employed similar techniques, despite the fact that differences between geographical isolates may reflect environmental rather than genetically controlled factors. Studies of the prevalence and transmission of animal, human and zoonotic filarids, so important for vector identification and control, has lead to the examination of filarial parasites at the genetic level. Genomic DNA libraries are being contructed and screened for clones which are species specific. From this work, DNA probes which can accurately enumerate larval stages in vector squash preparations, and monoclonal antibodies specific for defined filarial antigens, are being prepared. The nucleotide sequences of rRNA are also being defined. The application of these technologies to the study of filarial parasites and their vectors, promises to not only allow the construction of accurate phylogetetic trees, but also to provide the data necessary for the identification and control of the vectors of filarial pathogens of animal and man.
Antigenaemia as an Indicators of Filarial Endemicity
F. Partono; R. Maizels; Purnomo; E. Sartono.
Abstract :
This a report of 1-years evaluation of chemotherapeutic intervention in an area of Indonesia endemic for lymphatic filariasis. Control measures were initiated in 1977 by parasite control, informal health education, and community participation at the village level, well in accord with the WHO-concept of health for all. Diethylcarbamazine (DEC) was mass distributed in 1977 and 1988, and selectivelly distributed in 1978, 1979, 1981, and 1982 to those who were micro-filaraemic to DEC treatments, those with a history of adenolymphangitis over the previous one year period, and to all new comers. In addition, each villager with acute symptoms of adenolymphangitis was immediately treated with a single course of 300 mg DEC for 10 days. No intervention measures were taken between 1982 to 1988, and no attempt was taken to control the vector or to restrict movement between controlled and uncontrolled areas during the whole studies. With these measures, the microfilaria (mf) rate decreased from 30% to 0%, the adenolymphangitis rate from 46% to 11%, and the elephantiasis rate from 35% to 3%. The abatement of acute and chronic filarial symptoms over the study period and the disappearance of microfilararemia in the community are pointing towards the possibility of aredicating the partasite from the community. To test this hypothesis, serum samples were rested for circulating filarial antigen by a two-site antigen capture assay employing anti-phosphorylcholine monoclonal antibodies. There was a shsrp fall in circulating antigenaemia, demonstrating that infection has either been eliminated from nearly all villages, or that intensity of infection is now undetectably low. We feel that antigenaemia can be used as an indicator of filarial endemicity.
A Field Study Using the Polymerase Chain Reaction (PCR) to Screen for Brugia Microfilariae in Hman and Animal Blood
Janet Glover; Steven A. Williams; Susanne Szabo; David Landry; Larry A. Mc.Reynolds; Taniawati Supali; Felix Partono.
Abstract :
Blood samples from 43 humans and 14 cats positive with Brugia microfilariae were analyazed in a field study in Tanjung Pinang, Indonesia. The Study used the polymerase chain reaction (PCR) to compare the sensitivity of radioactive and biotinylated species-specific oligonuleotide probes. The cloning cahir acterization of the Hha I repeat DNA family found in filarial parasites of the genus Brugia, and the development of species-specific probes for B. malayi and B. pahangi based on these repeats has been described elsewhere (PNAS USA 83: 797-801). Mol. Biochem Parasitol. 28: 163-170). The use of radioisotopes for labeling DNA probes is both expensive and inconvement. To replece these probes, biotinylated DNA probes have been designed for non-radiactive detection of B. malayi and B. pahangi. These oligonucleotide probes have long tails of biotinylated uridine residues added to their 5' end. As little as 100 pg of Brugia DNA can be detected on dot blot with these probes. Detection of the probes is based on an avidin-alkaline phosphatese colorimetric assay. In order to distinguish between infected from uninfected individuals, it is necessary to detect the amount of DNA in one microfilaria (about 60 pg).
The polymerase chain reaction (PCR) is a procedure in which a small amount of of DNA can be amplified up to 1 million-fod. A part of each sample in this study was PCR amplified and compared with unamplified portion using both the radioactive and biotinylated DNA probe.
The PCR amplified samples were accurately identified by both the radioactive and biotinylated B. malayi and B. pahangi probes. Even samples with as few as two microfilariae per 100ul of blood were easily detetcted. The samples that were not PCR amplified were accurately identified after only long exposure (greater than one week) to the radioactive probes. The biotinylated probes, were not sensitive enough for accurate identification of the non-PCR amplified samples. The polymerase chain reaction is, therefore, a promising new tool for enhancing the sensitivity of parasite detection assays based on DNA probes. This will be especially important in designing assay based on non-radioative DNA probes.
Detection of Brugia malayi Infected Mosquitoes with Species Specific DNA Probe pBm 15, in Riau, Indonesia
L. Kurniawan; Sjahrial Harun; Basundari Sri Utami; B.K.L. Sim; W.F. Piessens; S. Atmosoedjono
Abstract :
Species specific DNA probe (pBm 15) was used in a field area where 2 filarial infections coexist : B. malayi in man and B. pahangi in cats. In our laboratory in Jakarta, this DNA probe proved to be sensitive enough to detect 500 ng DNA. One to two infective larvae of B. malayi could be detected with ease. This DNA probe did not react with infective larvae of wuchereria bancrofti, B. pahangi, and Dirofilaria spp.
Non specific binding caused by undefined mosquito components was overcome with proteinase K and chitinase treatment. This additional step, made it possible for whole body mosquitoes to be squashed directly onto nitrocellulose paper
A comparative study of experimental infections of laboratory bred mosquitoes infected with B. malayi, showed no difference in infection rate between the group examined by dissection or by DNA probing.
Mosquitoes which are vectors in Riau were colected and fed on microfilaremic patients of Riau. The set of mosquitoes were tested in parallel with mosquitoes infected with B. pahangi form cats. All fed mosquitoes were tested after 10-12 days. Only mosquitoes infected with B. malayi in the assay.
This study shows a success in appliying the DNA probe technique in Jakarta. Further application in the field should be encouraged, with some modification of the DNA probing technique, for cheaper and easier implementation.
Detection of Brugia malayi and Brugia pahangi Parasites by Biotinlabeled DNA Probes
T. Supali; S.A. Williams; C.B. Poole; J. Glover; L.A. McReynold; F. Partono.
Abstract :
Morphologically, the larvae of Brugian parasites are difficult to differentiate by conventional mmethods. Recently, radiocative labeled DNA probes2,3 have been developed to distinguish the larvae of these parasites. However, these probes have a short shelf-life and are hazardous to the users. Two oligonucleotide DNA probes have been tested, one is specific for B. malayi and the other specific for B. pahangi. They were each labeld with Biotin in three different ways by using : a one-tailed 30mer biotinylated uridine residues, a two-tailed 30mer biotinylated uridine-thymidine residues. The dot blot assays were tested at various tempereture (300C-800C) using different concentration of parasite DNAs (12,8 ng-0,1 ng). Our preliminary results indicated that the sensitivity and specificity of the biotinylated DNA probes, with a two-tailed 45mer biotinylated residues, were highly acceptable for field use.
The Tubulin Gene of D.immitis
Nur Asikin.
Abstract :
A cDNA clone of D.immitis in the expression vector/gtll gave positive reaction when screened with a specific antisera raised against sea urchintubulin. The clone contained a 342 base pair insertion; the DNA sequence is about 75% homoglous to tubulin sequences of other organisms, whereas the deduced amino acid sequences is about 90% homologous.The Southern blot analysis inicates a unique restriction map for the DNA surrounding the tubulin clone sugessting the presence of only one-tubulin gene in D.immitis.
Use of Melocular Genetic Engineering in the Study of Animal Parasites and Their Vectors
Philip T. Loverde.
Abstract :
Molecular genetics coupled with advences in immunology and parasite culture has become a powerful tool to study animal parasites and their vectors. Recombinant DNA techniques allow one to identify individual genes of DNA probes, amplify the nucleic acid of interest, and use this material to study : the role of the gene product in the biology of the organism; the evolution of parasites and their hosts; heterogeneity between species and within species; toxonomy and develop refined toxonomic tolls; the immunology and biochemistry of host-parasite interactions; identification if specific cells or tissues that produce gene products; cytogenetics and localization of genes on chromosome in the study animal parasites and their vectors will be presented.
Important Trematodes in Man in Indonesia
Pinardi Hadidjaja
Abstract :
Trematode infections are considered to be a non public health problem in Indonesia, with the exception of the two species namely Schistosoma japonicum and Fasciolopsis buski which are still assumed to be the most important trematodes of Indonesia. Other trematode infections reported occasionally were caused by : Echinostoma spp, Haplorchis yokogawai, Haplorchis taichui, Paralecithodendrium molenkampi, Phaneropsolus bonnei and Plachiorchis javensis.
Schistosoma japonicum infection has been known to be endemic among the inhabitants of Lake Lindu since 1937. Surveys carried out since that time revealed prevalence rates which varied from 8-55%. Further studies proved that the species at Lindu were similar to the species found in Asia. Resurvey initiated in 1971 gave almost a similar prevalence rate followed by the discovery of the molluscan intermediate host namely Oncomelania hupensis lindoensis.
In 1972 a new focus of schistosomiasis was discovered at Napu Valley. Other survey did not reveal any other endemic foci. The result of a biological study carried out since 1976 reconfirmed the diagnosis of the species. Clinical study showed taht the significant signs and symptoms of schistosomiasis found among the Lindu inhabitants were : dermatitis, diarrhea, dysentry, abdominal pain, nausea and vomiting, loss of appetite, weakness, distention of the abdomen, melaena, hepatomegaly and splenomegaly.
Control of schistosomiasis was initiated in 1981, using praziquantel for mass teratment. Twelve cycles of tretament at Lake Lindu area and 10 cycles at Napu Valley resulted in a reduction of overall prevalence rate from 15,80% at Lindu area in 1981 to 1,14% in 1987 whereas in Napu Valley the reduction was from 35,8% in 1982 to 1,00% in 1987.
Fasciolopsis buski infection was first reported in 1982 from the Regency of Babirik, Hulu Sungai Utara in South Kalimantan Province. The following survey in that area in 1986 revealed a prevalence rate of 27,0%. Clinical examination showed that some of the complaints were : diarrhea, poor appetite, mild abdominal colic or burning sensation, vomiting and fever. Physical examination revealed emaciation, anemia, distented abdomen, ascites, and jaundice. Another survey in four other adjacent villages showed prevalence rates that varied from 0,00%-68,3%. The epidemiological survey carried out recentlu did not reveal the intermidiate hosts except only two metacercariae which could not be identified.
Current Technological Approaches to the Study of Trematodes
W.P. Carney
Abstract :
Advances in biology sciences over the past decade have provided additional technology for dealing with trematode parasitoses. Although trematode disease of man and animals in Indonesia are assentially the same as they were ten years ago, scientific methods available to deal with them have improved significantly. One can now exploit the innate ability of cells to replicate and produce biological products upon demend, manipulate the genetic make up of an organism, biologically or synthetically manufacture peptides and rationally develop drugs that target iidiosyncrasies of parasites at the cellular and molecular levels. Further, one can now manage and analyze massive amounts of biological dtat using desk top computers. These new biological techniques and the computing ability to interpret the data generated provide parasitologists in Indonesia and elsewhere with the ability to document the economic and public health impact of trematode parasitoses and to develop new strategies and reagents for diagnosing, treating, preventing and controlling the diseases they cause. In addition, biotechnology offes university scientists and their students with additional oppertunities to investigate basic and esoteric aspects of host-parasite interrelationship that are such an intriguing aspect of biology.
Molecular Approach to the Study of Trematode Parasites : The Blood Fluke
Philip T. LaVerde
Abstract :
One important aspect of reproductive development in trematode parasites in the formation of a hardened aggshell which allows the zygote to develop into a iracidium in a hostile environment. The miracidium then can transfer the germline from the vertebrate host to sanil intermediate host. Schistosome parasites, unlike other trematodes, have separate sexes and female reproductive development is known to depend on the presence of a male parasite. These facts make the blood flukes ideal material to study the mechanisms that underlie female reproductive development and eggshell formation. We reasoned that the morphological and biochemical differences between the male and female must be reflected at the molecular level in the differential expression of sexually regulated genes. Radioactive single stranded cDNA was first transcribed from female RNA; and then sequences common to both male and female were removed by hybridization to an excess of male RNA. This probe was used to screen a cDNA library made from mRNA of adult worm paris. One hybridizing clone, pSMf 61-46, was shown to correspond to a 0,9kilobase mRNA that is present only in mature female worms and is not detectable in female schistosomes from single-sex infections, in male worm or in eggs. Thus expression of the gene was female \-specific.During normal bisexual infection this mRNA is first detected 28 days after infection (the time of worm pairing) and increases to a high level at 35 days postinfection, coinciding with egg production. Thus the temporal expression of the gene was dependent on paining with male worm. The nucleotide sequence of the shows and open reading frame that encodes q 16 kDA polypeptide thhat show strong homology with eggshell proteins on insects. A second female-specific cDNA clone, F-4, represents a 1,6 kilobase mRNA whose expression is also corellated with worm pairing and subsequent egg production, encodes a different putative eggshell component of 44 kDA. The amino acid composition of the 16 kDA and 44 kDA polypeptiders show a strong correlation with the actual animo acid composition of the schistosome eggshell. Thus these two polypeptides appear to the major components of the schistosome eggshell. Analysis of the genomic arrangement of the eggshell genes show that p16 is represented by 5 gene copies and p48 is represented by 2-5 copies. The eggshell genes are expressed in the vetelline cell as recently demonstrated by in hybridization and immunocytochemical localization. The eggshell genes are being expressed in bacteria. The gene products will be used to study the biochemistry of eggshell formation.
The Application of a Numerical Method for Taxonomy from Isoenzymatical Result : Coefficient of Parental Correlation Between Schistosoma Species
Sri Subekti Bendryman Soedjoko
Abstract :
Using enzyme characters by starch gel electrophoresis, we have applied the method of Numericla Taxonomy to the Schistosomes isolate/stocks.
Seven isoenzymes (Phosphoglucomutase = PGM; Phospho glucose isomerase = PGI; Hexokinase = HK; Mannose phosphate isomerase = MPI; Alcaline phosphatase = ALP; Malic enzyme = ME and Malate deshygenase = MDH) of 15 isolats/stocks are examined.
Twenty six electromorphs, corresponding to equivalent number of isoenzymes were used as Operational Toxonomy Unit (OTU) and compared pairwise, using these indice and it forms the basis for the toxonomic scheme elaborated. The final relationships are exhibited in the egglamerative dendogram, constructed using complete linkage.
The sparation into phenons is confirmed by correspondece analysis. It is concluded that the original lines fall into four groups correspondings to the complexes Schistosoma mansoni, S. bovis, S. curasonni and S. haematobium. The phenons are recognised by Numerical Taxonomy, can be equated with the taxa of traditional systematics.
The Medically Important Molluscs of Indonesia
Machfudz Djajasasmita.
Abstract :
At present in Indonesia 32 species of non-marine mollucsc which belong to 15 families have been known to be potentially of medicalor veterinary importance, since they are suspected to be capable in transmitting human and animal diseases. The brackish water family Potamididae comprises of 1 species; whereas the freshwater snails are : Viviparidae (3 sp), Ampullariidae (1 sp), Pomatiopsidae (1 sp), Thiaridae (7 sp), Lymnaeidae (1 sp), Planorbidae (5 sp); freshwater bivalve are : Corbiculidae (4 sp); land sanils are : Subulinidae (2 sp), Achatinidae (1 sp) and Bradybaenidae (1 sp); land slug : Veronicelidae (2 sp), Philomycidae (1 sp) and Limacidae (1 sp), all are common species which can be found in the vicinity of human habitation (ponds, rice-field, ditches, gardens etc).
The parasitological studies on these molluscs are rather limited, only 9 species have been studies and confirmed to be the intermediate host of the blood fluke Schitosoma japonica in Central Sulawesi and Flores have been found to be harbouring the larvae of the nematode Angiostrongylus cantonensis, the causative agent of eosinophilic meningoencephalitis; Bellamya rudepelis, Gyraulus sarasinorum and Carbicula lindoensis were recorded as the intermediate host of the intestinal fluke Echinostoma lindoensis in Central Sulawesi; Lymnaea rubiginosa plays an important role in the life-cycle of the cattle liver fluke Fasciola gigantica and F. hepatica, which may reduce the national meat production; and Digoniostoma truncatum from Bali has been recorded naturally infected with radiae and cercariae of Paramphistoma sp., the causative agent of the of the fatal paramphistomiasis of cattle. Gyraulus convexiusculus is considered to be the most likely first intermidiate host of Fasciolopsis buski infection in human in South Kalimantan. AMong these moluscan species only Oncomelania h. lindoensis and A, fulica are the most studies species (life-cycle, transmission dynamics, control and ecological aspects), whereas the work on the other species are mostly emphasized on the taxonomic aspect.
Biotechnology in the Study of Molluscs
Christopher J. Bayne
Abstract :
The optimization of conditions and media for the in vitro culture of molluscan cells have yielded advences such that primary cultures can now be maintained for weeks of months. With one exception, molluscan cells have proven refractory to the establishment of continous cell lines. Knowledge derived from these studies has helped investigators maintain and grow intramolluscan parasites in vitro.
Recent advences in cell fusion, cell cloning and transfection of oncogenes have yet to be applied in molluscan cell culture. Once established, future cell lines will permit a great variety of studies including the control of gene expression, post-transcriptional processing including glycosylation, and the production in quantity of molluscan factors such as lectins, hormoned, immunogenic macromolecules and others of interest. In addition these lines will also facilatate screening for molluscan viruses and other cellualr pathogens, and the propagation of these putative screening for molluscan viruses and other cellular pathogens, and the propagation o f these putative agents of biological control. Such cell lines would also permit the study of molecular mechanism of drug and hormone actions, and the rational design of drugs.
Molecular genetic protocals have been restrited so far to studies of evolution and systematics within molluscan taxa. Both RNA and DNA probes are being developed from ribosomal, mitochondrial, and nuclear compartments. Both driect sequencing and restriction length polymorphisms detected using endonucleases are yielding useful data.
Importan Protozoan Parasites in Indonesia
Srisasi Gandahusada
Abstract :
The most important protozoan parasites in Indonesia are the malaria parasites, Toxoplasma gondii and Entamoeba histolytica.
After the second world war the rseidual insecticides and effective antimalarial drugs were used in the control of malaria. After development of resistance among mosquitoes to insecticides, the Malaria Control Programme was switches over to the Malaria Eradication Programme. Malaria incidence dropped heavily. However, due to the quick development of vector resistance and financial limitations, malaria came back and so did the Malaria Control Programme. P. falciparum and P. vivax are tho most common species in Indonesia. Important vectors are An. sundaicus, An. aconitus, An. maculatus, An. hyrcanus group An. balabcensis, An. farauti ect, An. sundaicus, and An. aconitus have developed resistance to DDT and Dieldrin in Java.
In 1959 the malaria Eradication Programme was started in Java, Bali and Lampung. In 1965 the API dropped to 0,15 per thousand. From 1966 onwards malaria transmission was on the increase, because spraying activities were slowed down, but dropped again from 1974 onwards by occasional residual house spraying with DDT or Fenitrothion, amalria survailance and treatment of malaria cases, resulting in an API of 0,18 per thousand in 1987. At present malaria is not transmitted in Jakarta and in capitals of the provinces and kabupatens, except in Irian Jaya, Nusa Tenggara Timur and one or two other provinces, but still exists in rural areas. The distribution of chloroquine resistant P. falciparum is patchy. Resistance is at the RI, RII, and RIII levels.
The main problems of malaria control are : the increasing development of resistance of the vector to insecticides, the change of An. aconitus from zoophili to anthropophili and from indoor to outdoor biting, the increasing resistance of P. falciparum to chloroquine, the shortage of skilled manpower and limitation of budget.
In indonesia many newborns with congenital anamolies are found. T. gondii as one of the causes, is widely spread in man and animal. The prevalence of Toxoplasma antibodies in man varies from 2% to 63%, in cats and other animals it can reach up 75%. Confirmed cases of congenital toxoplasmosis are reported. The diagnosis of toxoplasmosis in the Department of Parasitology, University of Indonesia is done with detection of specific IgM and IgG antibodies with ELISA. A test for antigenemia to get a rapid and direct diagnosis of active infection is not yet available. A suitable toxoplasma vaccine to prevent toxoplasmosis would be desirable. E. histolyca infection is endemic throughout the archipelago. The prevalence rates are 18% to 25%. Extraintestinal infection mostly occurs in the liver. Pulmonary amebiasis is occasionally found. Medication with metronidazole has obtained good result. Diagnosis of extraintestinal amebiasis in our laboratory is by immunodiffusion test, which is not capable to differentiate active infection from infection in the past. A more accurate diagnosis would be the use monoclonal antiboides to detect antigens.
Important Protozoan Diseases of Animals in Indonesia ( a review)
Soeprapto Soekardono.
Abstract :
An account on important protozoan diseases mostly with obvious clinical symptoms are empashized and their current status reveiwed.
Those diseases are surra, trichomonosis in cattle, babesiosis, anaplasmosis, theileriosis, leucocytozoonosis in chicken, and coccidiosis.
Toxoplasmosis, histomonosis, chicken malaria, balantidiosis and diseases coused by Giardia, Haemoproteus and Sarcocystis are not reveived because significant problems coused by these parasites considered important economically do not appear in Indonesia.
Biotechnology in the Study Protozoan Parasites
Donald E. Burgess.
Abstract :
Over the past decade extraordinary advances have been made in the study of protozoan parasites. Particular progress has occured in areas such as cultivation of protozoan parasites, immunobiology of protozoan parasitic diseases, the biochemistry of protozoa and molecular genetics of these organisme.
The application of sophisticated culture, monoclonal antibody and recombinant DNA technologies has resulted in elucidation of many of the biochemical and molecular bases of such phenomena as antigenic variation in African trypanosomas, the autoimmune basis of the pathogenesis of Changes' disease, protective immunity in malaria and parasite evasion of host defense mecahnisms. As. a result a new generation of diagnostic procedures have to provide more accurate detection of prozoan infections and thus improved epidemiological information. Vigorous vaccine development effeort are underway which will lead to molecularly defined vaccines taylored to specific applications and will provide new weapons to combat protozoan diseases. Perhaps most importantly the mollecular bases of host-parasite interactions are being setablished and will allow inditificaition of unique biochemical aspects to the biology of protozoa thereby revelaing appropriate targets for development of vaccines, accurate detection procedures and more efficacious chemotherapeutic agents.
Detection of Plasmodium Falciparum in Blood Using DNA Probe, pPF 14
Syahrial Harun; Liliana Kurniawan; Robert H. Barker, Jr; Dyan F. Wirth.
Abstract :
A DNA probe pPF14, which is species specific to P. falciparum have been used to identify malaria species. The blood specimens were preteated before spotting onto nitrocellulose filter to eliminate non specific hybridization of DNA to blood components.
In this study, the DNA probe pPF14 was used dfor detection of P. falciparum in blood speciments spotted directly (a simple preparation of sample) onto nitrocellulose filter. Ninety two blood specimens from individuals with fever living in a P. falciparum endemic area were collected at the local Primary Health centre in Lampung. Blood was colected by finger prick technique using heparinized capillary tubes. Twenty microliter of blood was plotted directly onto nitrocellulose filter. extrcted, treated with proteinase K and tested with DNA probe pPF14. Another 20 microliter of blood was used for thick smear and examined microscopically. Nineteen out of 92 (20,7%) of the blood specimens were found to contain P. falciparum parasites by DNA probing and by thick smear. The densities of parasites ranged from 350 to 28,775 per microliter.
The DNA probing technique was succesfully done for the detection of p. falciparum in the laboratory. However, this technique seems not to be quite sensitive for parasitological survey (detects 40-50 parasites per microliter of blood); while using thick smear examination, 5-10 parasites per microliter of blood could be detected with confidence.
The important issue in this case is that no special treatment before sportting the blood onto nitrocellulose filter was needed to eliminate non-specific hybridization, when protease treatment was used. Collection of specimens would be easily done in the field.
Vector of Malaria and Filariasis in Indonesia
Hoedojo
Abstract :
Malaria at present is still one of the important mosquito-bornediseases in Indonesia. The disease is widespread all over the country and involves nearly all island. Sixteen Anopheles species have been reconfirmed as malaria vectorrs. They were distributed geographically as follows :
Coastal areas and lagoons ------- An. sundaicus and An. subpictus
Cultivated ricefields and swampy areas ------- An. aconitus, An.barbirostris, An. nigerrimus and An. sinensis.
Forest inland areas in shaded temporery
pools, muddy animal wallows and
hoof-prints ------ An. balabacensis, An. bancrofti, An. farauti,
An. koliensis, and An. punctulatus
Swamp forest edge in ditches with vege- ------ An. letifer and An. ludiowae
tation Hilly areas in seepages, streams and
clear moving water ------ An. flavirostris, An. maculatus, An. minimus.
The species (of most general importance if An. sundaicus, which is restricted) by its preference for brackish water and water and is prevalent in coastal areas of Java.
Their types in behaviour of An. sundaicus appear as follows :
- An. sundaicus in South Coast of Java in general.
This species is essentially anthropophilic, exophagic and rests outdoor. It shows susceptible to DDT. - An. sundaicus in Cilacap, Central Java
This mosquito is a pure anthropophilic form. It bites man in houses and outdoors, rests indoors and is known resistant to DDT. - An. sundaicus in Yogyakarta and Purworejo, Central Java.
This mosquito is a strong zoophilic species. It rests and prefers to bite outdoors and shows tolerance to DDT.
Human filariasis in Indonesia is the result of infection by three endemic species, nemmaly, Wuchereria bancrofti, Brugia malayi, and Brugia timori, W. bancrofti infection is found in both urbanand rural areas. Twenty species of mosquitoes are confirmed as filariasis vectors. The urban type bancroftian filariasis iss transmitted by Culex quinquefasciatus, whereas the rural trasnmitted mostly by Anopheles spp., such as An. aconitus and An. punctulatus complex. The periodic species of Mansonia transmit the subperiodic noctural B. malayi, b. timori which is distributed in the Eastern part of Indonesia (east Nusa Tenggara) is transmitted by An. barbirostris.
Some filariasis vectors suuch as An. aconitus and the An. punctulatus complex may function both as filariasis vector and malaria vector as well.
An. barbirostris with is confirmed as a vector of malaria in South Sulawesi, a vector of periodic noctumal malayan filariasis in Central Sulawesi and as the only vector of timorian filariasis in Timor and Flores, has to be studies further as it has two types of behaviourisitic appearance, namely :
- An. barbirostris in Java
This mosquito is an anthropozophilic species, feeds indoors and outdoors, and rest outdoors.
None is found to transmit mosquito-borne disease.
- An. barbirostris in Sulawesi and East Nusa Tanggara (outside Java).
This mosquito is a zooanthropophilic form, endo and exophagic, and rests outdoor.
It is confirmed as a vector of malaria, periodic nontural malayan filariasis and the only vector of timorian filariasis.
Multiplication of Dengue and Chikungunya Viruses in Aedes Mosquitoes
Soedarto Soekiman
Abstract :
Colonies of Eades aegypti (Surabaya strain) and Aedes albopictus (Malang strain) were studied to compare their susceptibility to oral infection with dengue type 3 and chikungunya viruses. Growth surves of dengue type 3 and chikungunya viruses. Electron microscopic observation of the salivary glands of A. aegypti and A. albopictus infected with chikungunya virus showed that this organ plays and important role in producing and maintaining high virus titers in these mosquitoes. The result suggest that both Aedes species are potentially important vectors on the transmission of dengue and chikungunya infection in Indonesia.
The Sporozoite Enzyme-Linked Immunosorbent Assay : Application in Malaria Epidemiology
Michael J. Bangs.
Abstract :
Recent biotechnological breakthroughs have led to the development of various methods for detection and identification of human pathogens in their vectors. Monoclonal antibodies produced against malaria sporozoite antigens have permitted the development of several sensitive, specific immunological tests (IFA, IRMA, ELISA). One of these, a two-site enzyme-linked immunoserbent assay (ELISA) has been developed as a useful epidemiological tool in the identification of malaria-infected mosquitoes. This method employs highly species specific monoclonal antibodies that recognize the repetitive immunodominant epitope of thee circumsporozoite (CS) protein. Monoclonal antibodies have been developed for all four species of human malaria. The key feture of the ELISA technique is the use of an enzyme indicaitors for an immunological reaction. The antigen capture or "sandwich" ELISA configuration uses the purified monoclonal boths as the solid phase and, conjgated to enzyme, as a marker for the presence of CS protein in a mosquito homogenate incubated in the wells of a microtitration plate. This technology has showen advantages over methods for epidemiological data colecction.Mosquitoes can cought, dried andstored until a time convenient for examination. The sporozoite rate by Plasmodium species can be identified easily, and combined with the man-biting rate provides the sporozoite rate, an important antomologic estimate of potential infective bite a person could expect over a given period of time. Presently, mosquitoes can be tested individually or pooled up to 20 anophelines. The assay is sensitive enough to detect 1 infected mosquito per pool or as few as 25 sporozoites per 50 ul of mosquito extract. Basic priciples and procedures are covered concerning solid substrate, adsorption to solid substrate, buffers and was solutions, conjugates and enzyme substrates. The advantages and limitations of thie technique in present malaria studies in discussed.
Seroepidemiologic Correlations in Malaria
Kevin Baird
Abstract :
Serologic measurements of humoral immunity have been used to estimate malaria transmission in endemic areas. The usual methods employ enzyme linked immunoserbent assays (ELISA) and immunofluorescent antibody tests a wide variety of malaria antigens. In theory, higher levels of antibody reflect higher levels of exposure to malaria antigen i.e., disease. However, one must cerefully consider variables such as endemicity, species of malaria present, age of the group examined, and the antigent/test array selected.
Withoud doing so, it is easy to draw erroneous conclusions. This presentation provides guidelines to selecting a test and antigen appropriate for deriving given epidemiologic conclusions from serologic surveys. Also, recent work on DNA probes of malaria and serum markers of cell-mediated immunity is context of epidemiologic measures of malaria transmission.
Plaque Assay of Newcastle Disease Virus
B. Sardjono
Abstract :
The newcastle disease virus (NDV) was isolated from a 3 months-old indigenous chicken (buras or kampung chicken) which showed clinical signs of Newcastle disease (ND). For viral isolation a small part of the spleen and lung were inoculated into 10 days-old embryonated chicken eggs. The physical characteristics of the isolate (A/120) were studied. The hemagglutination of chicken red blood cell showed slow elution, thermostability of hemagglutinin at 560 C was 120 minutes. The virus was able to agglutinate horse erythrocytes but not those of sheep. The biological characteristics on mean death time (MDT) of embryonated chicken egg and plaque morphology on chicken embryo fibroblast (CEF) primary cell cultures were studies. The MDT was 56 hours, the isolate was velogenic NDV. Three were three different plaque morphologies on CEF : 2 mm clear plaques, 1 mm clear plaques, and minute clear plaques which were visible only with microscopic examination.
The Murine Cell Surface Antigen PC-1 as a Marker for Plasma Cell
Alida R. Harahap; James W. Gooding
Abstract :
The murine plasma cell antigen PC-1 is a cell surface glycoprotein which is disulfic-bond homodimer of Mr 115 kD. It is expressed in large amounts of neoplastic plasma cell and on the majority of haemolytic plaque-forming cell1. It is also found in non-lymphoid tissues.1,2 In this paper a report of detailed distribution of PC-1 in a variety of normal lymphoid tissues using radioiodinated IR-518 monoclonal anti-PC-1 antibody as a probe is given, followed by autoradiography. Bbecause the number of plasma cells in normal condition is very small, mice infected with Mesocestoides corti have been used. These M. corti infected mice have high levels of IgGI in their serum (Mitchell et al, 1977). Hence, it was expected that the number of plasma cells may also be increased.
Most small lymphocytes are negative of weakly positive. Cells that had morphology of typical plasma cells were mostly positive. The large immunoglobulin-containing cells found in peritoneum of M. corti infected mice were strongly positive for PC-1. These result show that PC-1 antigen is expressed on normal plasma cells, the normal counterpatr of myeloma cells. Therefore it can be used as a plasma surface marker.
Anti-NP Immunoglobulin Gene : Somatic Hypermutation Does not Directly Influence its Specificity Change from Heteroclitic to Homoclitic
Amin Seobandrio W.K.
Abstract :
It is generally accepted that in the maturation of an immune response, the quality of the antibody, as well as the quantity, will improve with time after immunization. In the mouse system, immuno- globulin obtained in the first days after immunization with NP-CGG showed greater specificity to NIP-CGG (Heteroclitic specificity). While the specificity to the immunizing antigen (homoclitic specificity) was recovered few weeks later. Intensive study on its immunoglobulin struucture and the sequences of its mRNA revealed that this change of specificity was due to the swith from kappa to lambda light chain rather than to the occurence of somatic hypermutation in the immunoglobulin gene.
In this paper, we describe the contribution of either somatic hypermuitation or immunoglobulin light chain class switch to the change of specificity from heteroclitic to homoclitic.
Tailoring Immunotoxin as Anticancer Drug
Santoso Cornain; Nur Asikin; Hambali; Nur Huda; Ali Baziad; Enud J. Suryana; Farid Azis.
Abstract :
The conventional tretaments for cancer have been considered unsatisfatory, with limited efficiency in terms of discriminative cancer cell adverse reaction against the normal compartments, a number of immunological approaches had been implemented. Since cancer cell could exhibit tumor specific antigen(s), a highly specific antibody could be used to direct any anticancer drug, biological agent or radioisotope selectively against the cancer cells and does not harm the normal cells. The specific antibody could be raised by immunization with purified tumor specific antigen (s). The biological agent could be obtained as toxin, either derived from bactria e.g. diphtheria toxin or derived from plants e.g. castor ricin, which could destroy and cancer cells after contacts.
A hybrid molecule constructed between antibody and toxin has been know as "immunotoxin". The selectivity of the antibody against a given tumor specific antigen could be increased bu using a monoclonal antibody, made by hybridoma technique and immunological engineering. Acoordingly, the efficiency of the destructive or killing effect of the toxin could be eventually increased by puriification technique, biochemical and genetic engineering.
In a preliminary study ricin from castor (Ricinuss communis) have been purified and separated into two protein fractions (RCAI = 12.000 dalton and RCA II = 60.000 dalton). The latter showed toxin property, and was tested in vitro bopth against normal cells and against cancer cells. In the microcytotoxicity assay the ricin showed both the short term and the long term killing effect as measured after 1, 4, 16 and 24 hours. The killing effect against cancer cells was stronger as compared to that against normal cells. The acute or short term effect was observed at lower concentration of ricin (10-6 and 10-12 g/ml) after 1 and 4 hours contacts. The long term effect resulted in 90% and nearly 100% cytotoxicity in higher concentration of ricin. Further development of the immunotoxin are in progress. Various aspects dealing with technical problems and clinical aplications will be discussed.
Opprtunities in the United States for Training in Biotechnology for Parasitic Disease Investigations
W.P. Carney
Abstract :
In the United States there are academic, governmental and industrial training opportunities in biotechnology that are applicable to the study of parasitic disease.
Academic opportunities are the most plentiful. At least 130 universities in the United States have training programs in which current biotechnologies are being used to investigate parasites of economic or public health importance. Further, there are at least 70 centers of biotechnology in the United States. In these centers, many of which are located on university campuses, new biological techniques are being applied to both basic and applied research projects.
The united States government administers more than 20 research programe. These programs are conducted in or supported with resources of the following agencies: the Departments of Agriculture, Health and Human Services, Defence, and Interior, the Agency for International Development, The National Science Foundation, the National Academy of Sciences, and Environmental protection Agency.
Coomercial firms using biotechniological methods for production of biologicals and other reagents, provide and oppotunity for on-the-job training and experience in the application of these new technologies to parasites diseases of major economic or public health importance.
Soil-Transmitted Helminths in Yogyakarta
Noerhayati Soeripto; Dulyachman; Sri Sumarni; Sutarti.
Abstract :
Infections by soil-transmitted helminths in still a public health problem in Indonesia : Ascaris lumbricoides, Trichuris trichiura and the hookworms -mainly Necator americans- were the most encountered.
The rural villages near Yogyakarta in Central Java, Indonesia, werestudied for the endemicity of the worms. The population in the two villages were around 1200 people and regarded the same in respect of low socio-economic level and low educational level, differen occupations were applied in the villages.
In the villages of Kasongan the main occupationa wass morking in home industries making clay handicrafts such as ceramics and potteries, with bare hands. The materials were clay and sand, obtained from the soil surrounding the houses and from the riverbanks nearby. Environmental sanitation was very poor : most of the people had no latrines, defecation occured in the field or in the vicinity of the river. The climate was hot and humid. The village of Godehan was inhabited mostly by farmers, who also raised fish in open fishponds in the gardens or in the ricefields. Most of the houses did not have a latrine and defecation occurred in the fishponds or in the surroundings of the liver.
Faecel examinations were done by Kato's thick smear and modified Harada Mori culture technique. Soil samples were also taken from the several places and examined for A. lumbricoides eggs. The prevalence rates of the soil transmitted helminths in Kasongan village were significantly higher than those in Godehan village.
A. lumbricoides infections were 50,8% and 11,5%; T. trichiura infection were 82,6% and 36,6% and N. americanus infections were 76,5% and 25,3%, in Kasongan and Godehan respectivel. Soil samples were found positive for A. lumbricoides eggs in 28,3 of 538 soil samples in Kasongan and in 2,7% of 183 samples in Godehan.
The result indicated, that perhaps occupation was a factor for the significant differences in endemicity in the two villages.
Hybridoma Production Using Immunee Lymphocytes Against Brugia malayi Antigen with Myeloma Cells
Soeyoko; Noerhajati S; Abdul S. Safro.
Abstract :
Wuchereria bancrofti, Brugia malayi, and Brugia timori are the causative agents of lymphatic filariasis in Indonesia, but in some endemic areas, Brugia malayi is the most commonly found. Diagnosis of filariasis is normally based on clinical and parasitological examinations, but both have limitations. Therefore now the immunological examination plays an important role in the diagnosis of filariasis. The discovery of monoclonal antibodies recently may probably give a firm scientific basis in immunology and add a new dimension to the effort of developing a specific and sensitive immunological test for various stage filarial infection.
In this study, the production of hybridoma cells to develop monoclonal antibodies against B. malayi integrated a number of techniques : preparations of B. malayi surface antigen, immune lymphocyte cells, NSI myeloma cells and macrophage feeder layers, and a fusion of immune lymphocytes with myeloma cells.
The result of this study can be concluded in three points :
Protein analysis of the surface antigen was examined by Biureet and SDS-PAGE. A total of fourteen examniations were conducted by using 400 L3 for each experiment. Three were not detected by Biureet method, but showed five protein fractions by SDS-PAGE. The protein concentration of the surface L3 was varied from 85.0ug/ml to 769.23/ug/ml, with an average of 297.04/ug/ml.
The immunoreactivities of Balb/c mice antibodies to B. malayi L3 surface antigen were tested by ELISA and showed a gradual increase after four times immunization at two weeks interval. The optical density (OD) after four times immunizations was varied from 0.363 to 0.878 each mouse, where as the positive control sera OD was 0.570.
Hybridization using immune lymphocytics against B. malayi L3 surface antigen with myeloma cells yielde 60.41% cells and none of the producted monoclonal antibodies testes of ELISA.
Malaria in Central Java
Sugeng Juwono Mardihusodo.
Abstract :
Like in the Southeast Asian contries, malaria is still a public health problem in Indonesia. The province of Central Java is one of the most dense dense populated areas in Indonesia with an estimated 27 million populations in 1984.
Malaria inflicting human health in Central Java is primaliry caused by P. vivax, P. falsiparum and very rarely by P. malarie. Malaria cases are estimed by slide positivity rates (SPR), annual parasites incident (API) and annual Mood examination rates (ABER).
Small epidemics occured in several villages and the reasons for it were two fold, namely the occurence of DDT resistance by the main vector An. aconitus, An. sundaicus and An. maculatus and the occurance of chlorooquin resistance by P. falsiparum. The control programme made adaptations by using Fenitrithion in highly endemic areas and adminsitration of Fansidar in chloroquin resistant cases of P. falsiparum. The figures of SPR and API for 1979 until 1985 were constantly above 1.00, ranging from 1,18-4,19 for SPR and 1,37 until 6,00 for API. Fortunately the figures were coming down from 1986 until 1988.
In the 1988 was reported 0,67 for SPR and 0,63 for SPR and 0,63 for API. Constant survailance and lterness in endemic "pockets" is obligatory to prevent uprising.
Isolation of Bacillus sphaericus and Related forms Pathogenic to Culex quinquwfasciatus
J. SItumorang; S. Yuwono; A. Romas; M.M.
Abstract :
Five hundred and forty nine isoaltes of Bacillus spp. Sporeforming bacteria were obtained from soil, water, and mosquito larval samples collected in various lacolities of Yogyakarta Special Territory, Central Java and East Java. Four isolates were toxie to Culex quinquefasciatus; the belong to the species of Bacillus sphaericus (ascession number 23A and 51C), Bacillus cereus (ascession number 142A), and Bacillus pumilus (ascession number 25C).
Human Parasitic Infections in Bali : A Review
Putu Sutisna
Abstract :
Parasitic infections in human in Bali are well documented, aspecially in the population who llived in rual areas : The most common infections are those of the soil-transmitted helminthiasis which are caused by Ascaris lumbricoides, Trichuris trichiura and hokworm showing prevalence rates of 40-95%, 25-90% and 20-70% respectively. Enterobius vermicularis prevalence rate has been reported to be 18 -53%. Taenniasis prevalence rate has been documented to be 0,8-23% in some villages, where Taenia saginata was found to be more prevalent than 7 Taenia solium, and this might be due to the eating habit of the Balinese people who consumed both pork and beef lawar.
Malaria is still found in Bali especially in regions along the coasts of some regencies, although generally the infection rate is low. The prevalnce rates of intestinal protozoa such as Giardia lamblia, Entamoeba histolytica, Balantidium coli have been occasionally reported in low percentages.
The Inceidance of parasitic Diseases inLivestock in Bali
N.A. Suratma.
Abstract :
The population of livestock in Bali has continuously increased from year. However, some problems are encountered with parasitic infections in livestock. Parasitic infections may be coused by worms, protozoa or ectoparasites.
In catlle, the most common infections are those coused by Oesophangostomun sp, Ostertagia sp, Haemonchus sp, Mecistocirrus sp, and Cooperia sp which is the most dominatn. Neoascaris vitulorum was reported to be as high as 29,1% in calves. Fascioliasis in cattle was found highly prevalent, between 34,9 to 56,7% and was coused by Fasciola gigantica. Allso Paramphistomun infection was reported to be highly prevalent (50,1%). In addition, Boophilus microplus was recorded as high as 36,9%.
In goat and sheep, the incidence of Haemonchus contortus was 27,7% and 53,6% respectively. Infection of Paramphistomun sp in goat was 9,27%. Concerming ectoparates, Sarcoptes scabiei was reported to be caiuse of detah of 67% of young goats and up to 11% older gats in Br. Penginuman, Gilimanuk Negara.
Parasitisc infections in pgs were caoused by Cysticercus tenuicollis (11%) and Ascaris suum (24,2%) and 21,1% showed Metastrongylus apri and also Sarcoptes scabiei was reported too be the couse of skin disease in pigs.
In poultry, parasitic infection were coused by Raillietina (96%), Heterakis gallinea (66,7%), Capillaria sp (6,6%), Ascardia galli (56,7%), Oxypirura mansoni (50%), Acuaria spiralis (13,3%) and Syngamus trachea (3,3%). Multiple infection are common.
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