Taxonomic evaluation of Babesia canis isolates with different pathogenicity to dogs

Oral Sessions I Parasitology

D-0115

International

TAXONOtvUC EVALUATION OF BABESL4 CAM.9 IsOIATT3.5 WITH DWFRRENT PATHOGENIClT’Y To DOGS

47 (SuppI.) (1998) 133-281

0-0117

139

PHYLOGENBTIC RELATIONSHIPS BETWEEN ONCOMELANIA

HUPENSIS

LINDOENSIS

SUBSPECIES OF ONCOMELANIA

AND OTHER

INFERRED FROM

MITOCHONDRIAL 12SrRNA GENE -ln&otc for Compntive Tro+al Wdicinc andPamsitotogy,Univexsityof Munich, Gamzny. -Institute for Parasitologyand Tro+l Vctainary hWicioc of cbe univaily of Berlin, Gmnany. and -De+mont of InfccriousDismseaand Trqical Medicine,Univaaity of Munich, Germany

Okmnoto M’, TjahaJa H S**, Pinardi H”, Agatsuma T*** *Institute of Experimental Animal Sciences, Osaka University Medical School. Osaka, Japan. **Department of Parasitology, Faculty of Medicine. University of Indonesia, Indonesia and ***Department of Bmresource Science, Obihim

lnB&uiac&,markcddiffaclvrs in pathogealicity and vexor specificitybdwecn diffolult isolatedhave bee” dwxibcd. llwec s”baIxcied have thereforebeenaoposod:B. canis rossi with UK highest pathogcnicity and transmitted by HaemaPhysal~&aclri. B. canis cads with a hi& pathogenicity

and the vector Dmnmuen~or

nridahu.

and B. rank

vogeli

being of low patbogenicity and transmitted by Rhipicephalw sanguineus. However, it is not known. whether these subspxies repnscnt different gm_. Thenfore the fmt and seco”d intemal transcribed spacers and the intervening 5.8s coding region of the rRNA gene were compared in 8 isolated of the 3 poshdated s”b+ecies. incloding 3 which bad previously been chanaaizcd for pathoge”icity and vector specificity and “sed as reference stmins for the deamiption of the 3 subspecies. Tbc gea_ dunmined by seqoe”ci”g segregated into 3 clearly disti& grwp with the 3 refenmce strains belongi”g to one group each. Diffedmcea betwoo isolates wilhio the group wm only smaI1 (l-22 mutaticnrs). However, the 3 mference stmim showed marked diffwith 181~tiansbe2voc.B.canircrmir~B.crmLvogrliand325bctwem B.~muiPDdCPfhof~ootbcrsubspacjcs.ThiJwasin(hcsamcordtr of magnitude a8 the diff-

of the supposed subapccies to the homologous

of a” equine B&win. B. cab& (31-345). These res”lta therefore SW the taxo”omic divisio” of B. conir at tix subspecies level. gm

portion

perhap

eveal at tbz. spxied

level.

llJetargelgcaleisalsousetbtfordiagnosticpurposesbecallsesalI96l msuictiom enzyme dig&s of PCR prOawts of the B. canis isolatea examined

distinct ptterru corresptmding to the 3 proposed sobspzcies. Therefore PCR a”d nxtrictian fmgme”t length polymorphism (hap) analysis allow a quick a”d prognostically important differentiation 01 #he subspacics directly from blood samples.

Insulted

in

O-0116

dearly

IMPACT OF MOLECULAR PHYLOGENETIC ANALYSES ON CLASSIFICATION AND TAXONOMY OF COCCIDIA WITH ISOSPORAN-TYPE OOCYSTS

Tenter AM -InstiM ti Pamsitologie, Tier&tliche

Hochschule Hannover, Germany

The group of coccidia with isosporan-type (dispolic, tetmzotc) oacysts cwrently comprises about 400 named species with very diverse life cycles Several attempts have been made to classify these coccidia baaed on phenotypic chxxters. However, co-“s with regard to what weight to assign to the diiTerent characters and which characters to “se for defining different taxa has not been reached. Thus, a total of four diffwent families, five subfamilies and ten genera are mrrently being used for the classification of these coccidia. Recently, phylogenetic analyses based on moledar data, especially on 18s rDNA sequences, have provided new insights Into the phylogeny of various coccidian taxa-‘. The major concltions of these analyses are: (1) Coccidia with isosporan-type oocysts, such as homoxenous spenes of the genus lsospora and heteroxenousspecies of the tissue cyst-forming genera Toxoplasma and Sarcocyslrs, are very closely related to each other. They form a monophyletic group, to the exclusion of homoxenous coccidia with different wcyst morphologles, such as Elmeria and Cyclosporo (2) At least two ddferent lineages extst within the cxxcidia with isospomn-type cucysts. (a) One lineage comprises homoxenous and heteroxenous Isospora sptxies as well as a clade of Toxoplavna and Neospora both of which form tissue cysts (b) Another hneage comprises all of the Sorcocysru species analysed to date, which are monophyletic but form clades that are defined by their host specificity The fin&“g that homoxenous (Isosporo) as well as fac”ltatively (Isospora, Toxoplasma) or obligatory (Sarcocyst~s) heteroxenous coccidia wth ~sosporantype oocysts are a closely related, monophyletic group has major implications for their classdlcation, because It suggests that they should be classified in the same taxon. I suggest that all genera wth this type of ooqst morphology should be classlfed in the famtly Isospridae Minchin, 1903, wtth Isosporo being 1t.stype genus. In this classification, the fanuly Elmenidae comprismg the so-called classical coccidia, which have homoxenous life cycles with endogenous. intracellular phases of merogony and gamogony but different myst morphologies, is the sister taxo” to the Isospwidae. ’ Te”terAh4, Johnson AM (1997): Adv. Paras~tol 39,69-139, *Jefhss AC, Schnitzler B, Heydo,,, AO, Iohnson AM, Tenter AM (1997> 1 Euk Muobiol 44, 388.392; ’ Carreno PA, Schmtzler BE, JetTriesAC, Tenter AM, Johnson AM, Barta JR i199S) J Euk Micrabml (in press)

University of Agriculture and Veterinary Medicine. Obihiro, Japan Species of the intermediate snail host is one of the most important criteria in classification of the Schistosoma group. Africa” schistosomes. the S. mansoni and S. haemnrobium

groups, develop in pulmonate snails of the

family Planorbidae (Subclass Pulmonata), while oriental schistosomes, the S. japonicum

group and S. sinensrum, utilize the snails of the genus

Oncom&nio and its related species (Family Pomatmpsldae. Subclass Pmsobranchia). We have exammed phylogenelic relationships of some snail hosts of oriental schistosomes using mitochondnal DNA sequences. and reported that tive isolates of 0. h. quadrasi formed a monophyletlc clade and other 4 subspecies (0. h. hupensis, 0. h. nosophora. 0. h. fomwsana and 0. h. chiu’) were

closely related to each other (Parasitology International. 46 Suppl.

98.1997). In the present study, we examined paniai sequences from mitochondrial IZSrRNA gene (364368bp) of 0. hupensis lindoensis, the intermediate host of S. japonicum in Sulawes~ Is.. Indonesia. and compared it with those of other Oncomelania

species to infer Its phylogenetic powion.

Percentage nucleotlde differences between 0. hupensis subspecies m&ding 0. hupensis lindoensis

and 0. minima were about 9 to 12%. Computer analysis

for consuucting a phylogenetic tree showed that 0 hupenris lindoensis was fallen into one of three highly dwergent clades. An imphcated evolutionary relation among the snail hosts IS &cussed.

O-0118

ISOLATION. IDENTIFICATION AND COMPARISON OF “FREE-LIVING” AMEBAE FROM EYE INFECI’ION AND THE ENVlRONMENT

S.&n&i Z*, Endo T**, Yagita K **, V&h M***. V.&b I’, Nagy E’ *Dept. of ClinicalMicrobiology,AlbertSzent-GyOrgyi MedicalUniversity, Szeged. Hungary; *‘Dept. ofParasitology, National Inslitute of Illrechous Diseases, Tokyo, Japan; ***Dept. of Ophthalmology, Albert Szent-Gy(irgyi Medical University, &god. Hungary Amphimic small amebae are capable for existmg both in “free-living”and in “parasitic” form. Successful differential diagnosis and appropriate therapy of human infections (keratitis, encephalitis)causedby “frecliving” amebae (such as Acanthamoeba, Noegferia) depend on the precise laboratory identification of them. The growing use qf contact Imses increases the number of persons at risk of Acnnthamocba keratitis in Hungary, too. An Acanthamoeba sp. has been isolatedby us on agar plates covered with E. coli from the commercial contact lens cleaning and disinfectingsolution,as well as from the comeal scraping of a patient with severeeye soreness.Morphologically, this isolate (Cl) was indistinguishable from the Acanthamoeba strains isolatedfrom moss (MO) or from the Ri+e.r Danube (Dun). Using restriction fragment lengthprotile (RFLP) analysisof mitochondrial DNA (mtDNA), the pathogenic strain (Cl) and the strains isolated from the environment were found to be similar to the acanthamoebaeisolated from 16 human patients in differmt countries.As any strains of Acanthamoeba may have pathogenicpotential, the mtDNA RFLP phenotype may be a useful aid in the taxonomic classificationof “free-living”smebae speciesand probably also in the determinatiw of their pathogmic potential.Thus, a systemiccocperati% study was startedfor the isolation, identificationand determinationof the possiblepathogenicityof the fauna of the “free-living” amebae in Hungary, with specialrefermce to the sites of human recreation, such as natural waters, hot springs and swimmingpools. During our studieson the comparim of the RFLP of the isolates by the use of restriction endonucleasedigestion of mtDNA, we obserwd some extra bands in an axenically grown Acnnrhumoebo sp. isolated from the basin of a natural hot water spring. Electron microscopy revealed evidence for the multiplication of rod-shaped bacteria within the amebic cytoplasm The identification.characterization and determinationof a possiblepathogenicrole of that endosymbiontare in progress.