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And Toxoplasmosis in Jackass Penguins in South Africa

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And Toxoplasmosis in Jackass Penguins in South Africa IMMUNOLOGICAL SURVEY OF BABESIOSIS (BABESIA PEIRCEI) AND TOXOPLASMOSIS IN JACKASS PENGUINS IN SOUTH AFRICA GRACZYK T.K.', B1~OSSY J.].", SA DERS M.L. ', D UBEY J.P.···, PLOS A .. ••• & STOSKOPF M. K .. •••• Sununary : ReSlIlIle: E x-I1V\c n oN l~ lIrIUSATION D'Ar\'"TIGENE DE B ;IB£,'lA PH/Re El EN ELISA ET simoNi,cATIVlTli t'OUR 7 bxo l'l.ASMA GONIJfI DE SI'I-IENICUS was extracted from nucleated erythrocytes Babesia peircei of IJEMIiNSUS EN ArRIQUE D U SUD naturally infected Jackass penguin (Spheniscus demersus) from South Africo (SA). Babesia peircei glycoprotein·enriched fractions Babesia peircei a ele extra it d 'erythrocytes nue/fies p,ovenanl de Sphenicus demersus originoires d 'Afrique du Sud infectes were obto ined by conca navalin A-Sepharose affinity column natulellement. Des fractions de Babesia peircei enrichies en chromatogrophy and separated by sod ium dodecyl sulphate­ glycoproleines onl ele oblenues par chromatographie sur colonne polyacrylam ide gel electrophoresis (SDS-PAGE ). At least d 'alfinite concona valine A-Sephorose et separees par 14 protein bonds (9, 11, 13, 20, 22, 23, 24, 43, 62, 90, electrophorese en gel de polyacrylamide-dodecylsuJfale de sodium 120, 204, and 205 kDa) were observed, with the major protein (SOS'PAGE) Q uotorze bandes proleiques au minimum ont ete at 25 kDa. Blood samples of 191 adult S. demersus were tes ted observees (9, 1 I, 13, 20, 22, 23, 24, 43, 62, 90, 120, 204, by enzyme-linked immunosorbent assoy (ELISA) utilizing B. peircei et 205 Wa), 10 proleine ma;eure elant de 25 Wo. Des . glycoprotein-enriched frac ti ons to detect anti-B. peircei IgG. Th e prelevements sanguins provenont de 191 S. demersus adultes onl somples originoted from three groups of free-ranging penguins ele lesles par ELISA en utilisont les fractions de B. peircei enrichies (n = 1 10), 1 group of penguins (n = 66) which were rescued en glycoproteines pour deleclel les IgG dirigees con Ire B. peircei. after offshore oil-spill con tominations and rehabilitated ot th e South Les prelevements provenaient de trois groupes de manchols Africon Notional Foundation for the Conservation of Coastal Birds sauvages (n - 1 10), d 'un groupe de manchols (n = 66) ayanl ele (SANCCOB), and the final group from SANCCOB-resident secourus apres avoir ete contamines par une maree noire en mer penguins (n = 151. The overall B. peircei seropreva lence was et soignes d 10 Fondation Nationale Sud Africoine pour Ia 65 %, and the mean seroposi tivity ranged from 60 to 71 % Conservation des Oiseaux Lilforaux (SANCCOB), el d'un demier among the five penguin groups. The ELISA appecred to be specific groupe issu des manchals pensionnaires du SANCCOB (n = 15). for B. peircei IgG as tested against Haemoproleus columbae IgG La prevalence globale pour B. peircei etail de 65 %, et 10 and avian malaria (Plasmodium relictum, ond P. elongalum) IgG. seropositivile moyenne s'echelonnail de 60 d 7 I % parmi les cinq Toxoplasmo gondii antibody (Ab) were detected by th e d irect graupes de manchols. L'EUSA apparaissail specihque pour les agglutinati on tes t using kill ed T. gondii tachyzoites. All birds were IgG dirigees conlre B. peircei lorsque leslee pour les IgG contre selonegative for T. gonda Ab. The lock of T. gondij-positive Haemoproteus columbae el les IgG contre Ie paludisme aviaire penguins wos due to the appropria te so nitary conditi ons and anti­ (Plasmodium relictum, et P. elongatum) . les anlicorps (Ac) diriges Toxoplasma prevention procedures utilized by the SANCCOB. cantre Toxoplasma gondii ani ele delecles par Ie lest d 'agglulination directe utilisant des lachyzailes de T. gondii lues. KEY WORDS : Babesia peircei, Jackass penguins, African black·footed penguin , Tous les oiseaux etaient seronegatifs pour les Ac diriges conlre Spheniscus demersus, Toxoplosma gondii, ELISA, SDS·PAGE. T. gondii. I'absence de manchols posilifs pour T. gondii elait due aux condilions son ito ires appropriees et aux methodes de MOTS CLis : Babesia peilcei, manchot, Spheniscus demelSus, Toxoplasma gondii , ELISA, SDS·PAGE. prevenlion con Ire T. gondii ulilisees par Ie SANCCOB. • Johns H o rkin~ Unive rsity, School of Hygiene and Public Hea lth, The Jackass or Africa n black-footed penguin, Sphe­ De partmen t of Molecut ar Microbio logy and Immuno logy, niscus del17ersus, is an endangered species w ith decrea­ Baltimore, Maryland 21205, USA. sing populatio ns in its natural habitat on coasta l islets .. University of Cape Town Medica l School, Obsen'3tolY 7925, of South Africa (SA) (Crawford et aI. , 1990). Frequent Der aIlment of Anatomy and Cell Biology, Ca pe Town, South Afri GI. ... US Depa rtment of Agriculrure, Agriculture Resea rch Service, live­ offshore o il-spill contamination pr sents th e most stock and PoultlY Sciences Instit ute, Parasit e Biology and Eride­ devastating haza rds for existing wild p opulations. The miology Ltboratory, Beltsville, Maly land 20705, US A. spills debilitate, injure, and induce disease in penguins .... Percy FitzPatrick tnstitute of African O rnithology, University of (Erasmus el at., 1981; Kerley el al. , 1985). The disa­ Cape Town, Rondebosch 7700, South Africa. ••••• North C<lrolina State University, Enviro nmental Medicine bled penguins are rehabilitated in the rescue starion Consortium, College of Vete rinalY Medicine, Raleigh, Nortl1 Ca ro­ of the South Africa n National Foundation for the lina 27606, USA. Conservation of Coastal Birds (SANCCOB) in Milnerton Correspondence: Dr. Thaddeus K. Graczyk. Cape Town, SA, and released later T el (410) 614-4984 - Fax : (4 ] 0) 955-0105 - E- mai l : (33°55'5, ] 8°22'E), <[email protected]>. into the wild. Parasite, ]996, 4, 313-319 L_--1.M~em~02!i!!re!...-_--1------------- 3 13 GRACZYK T.K., BROSSY J.J. , SANDERS M.L., DUBEY JP., PLOS A. & STOSKOPF M.K. An inrrae rythrocytic piroplasm of free-ra nging (Brossy, 1992), air-dri ed, and stored on filter paper as S. delnersus originally described as Aegyptlanella sp. described by Graczyk et aL. (993). The samples ori­ (Coles, 1941) was reindentified as Babesia sp. (Ben­ ginated from three groups of free-ranging penguins nert et al , ] 992) and named Babesia peircei (Ea rl e from offshore islands (Dassen Island and Robben et al, 1993). Babesia peircei associated I abesiosis is Island) and mainland colony (Boulders, Simons Town), common and e ndemic in w ild S. demersus and contri­ from penguins rescued after offshore oil-spill conta­ butes to illness when associated with avian malaria minations and rehabilitated at SANCCOB, and from (Brossy, 1992 ; 1993) . The prevalence of B. peircei SANCCOB-reside nt penguins. The penguins were parasitemia in S. demersus rehabi litated at SANCCOB screened for blood parasites by Giemsa-stained thin vari ed from 11 to 15%, and was 4 % in free-ranging blood smears (Brossy, 1992), and a total of 10 ml of birds (Brossy, 1993). A putative Babesia sp. infection blood was collected from four penguins with the that ca used regenerative a naemia in ju venil e li ttle pen­ highest B. peil-cei parasitemia. Five thin blood smears guins (Eudyptula minor) in Australia has been reported were prepared and examined (Graczyk et aI. , 1994b) by CI<lrke & Knowles (993) from the blood sample, and 10 (30 pi each) blood Fatal Toxoplasma gondii associated toxoplasmosis was sa mples were air-dried and stored on filter paper. reported in capti ve Humboldt's penguins (s. bum­ Parasites were identified based on the description of bo/dli), Magella ni c penguins (S. magellanicus), and Ea rl e et aL. (993). The parasitemia was presented as S. demel'sus in orth America (Ratcliff & Worth, 1951). a mean % C:!: SD) of parasitized e rythrocytes pe r Another case of fatal toxoplasmosis, diagnosed histo­ 2,000 red blood cell s (RBC) derived from five counts. logicall y and immunologicall y, occurred in Tasmania, The blood for parasite extracti on was centrifuged Austra li a, in E. minor (Mason et al. , 1991) fed with 00,000 g, 10 min) at 4° C, and the plasma was stored sheep meat (Clarke & Knowles, 1993) by a person who at - 70° C. The packed RBC were subjected to two boarded many cats (Mason et aI., 1991) Toxoplasma freeze/ thaw (- 70° C/41° C) cycles and resuspended gondii infections are common in many domestic and in 10 ml of phosphate buffered saline (PBS) (pH 7.4). wild bird species (Dubey & Beatti e, 1988). Two hundred pi of N,N',N'-polyoxyethylene(lO)-N­ The ava il ability of penguins undergoing rehabilitation ta ll ow-l,3-diaminuprupane (EDT-20) (Sigma Chemical at SA CCOB and access to free-ranging birds offered Co., St. Louis, Missouri, USA), diluted 111 with deio­ an opportunity for serological screening of the birds for ni zed water, was added per 1.0 ml of [he preparation, pathogens. lt is crucial in penguin management to and the solution was incubated for 20 min at 4]0 C. assess accurately narurally occurring exposure to [he EDT-20 is a water-soluble cationic detergent th at tar­ pathogens in free-ranging S. demersus in order to deter­ gets lipoproteins and lipids without denaturing proteins mine the impact of exposure to oil contamination on (Schuttle & Ku la, 1990). The remainder of the protocol th penguin 's immunological competence, and to eva­ for parasite e>..1:racti on inclucting a 4-step Percoll gra­ luate the risk of releasing possibly infected I irds into dient centrifugation (4,000 g, 30 min) fo ll owed that of the w il d after rehabilitation. Av ail ability of the Graczyk et al. (1994c). The pellet containing the par­ S. demer.s-us-specific conjugate fo r use in enzyme-linked ticles of RBC and the supernata nt from the final para­ immunosorbent assay (ELISA) (Graczyk et al.
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