Oocyst shedding by green-winged-saltator (Saltator similis) in the diagnostic of coccidiosis and Isospora similisi n. sp. (Apicomplexa: Eimeriidae)

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1 Rev. Brs. Prsitol. Vet., Jboticbl, v. 22, n. 1, p , jn.-mr ISSN X (impresso) / ISSN (eletrônico) Full Article Oocyst shedding by green-winged-slttor (Slttor similis) in the dignostic of coccidiosis nd Isospor similisi n. sp. (Apicomplex: Eimeriide) Eliminção de oocistos por trinc-ferro-verddeiro (Slttor similis) no dignóstico d coccidiose e Isospor similisi n. sp. (Apicomplex: Eimeriide) Cleide Domingues Coelho 1 ; Bruno Pereir Berto 2 ; Dniel Medeiros Neves 3 ; Vinícius Modesto de Oliveir 3 ; Wlter Flusino 4 ; Crlos Wilson Gomes Lopes 4 * 1 Centro Universitário UNIABEU, Belford Roxo, RJ, Brsil 2 Deprtmento de Biologi Animl, Instituto de Biologi, Universidde Federl Rurl do Rio de Jneiro UFRRJ, Seropédic, RJ, Brsil 3 Horto Florestl Mário Xvier, Centro de Trigem de Animis Silvestres, Instituto Brsileiro do Meio Ambiente e dos Recursos Nturis Renováveis IBAMA, Ministério do Meio Ambiente MMA, Seropédic, RJ, Brsil 4 Deprtmento de Prsitologi Animl, Instituto de Veterinári, Universidde Federl Rurl do Rio de Jneiro UFRRJ, Seropédic, RJ, Brsil Received My 14, 2012 Accepted November 6, 2012 Abstrct Diurnl periodicity is phenomenon tht hs been observed in coccidin of Isospor prsites of psserines, which hve been eliminted gret number of oocysts t dusk. The objective of this study ws to evlute the occurrence of periodicity of oocysts presence in the green-winged-slttor Slttor similis, nd its use in the dignosis of coccidiosis in wild birds in cptivity. A totl of 220 fecl smples were collected from birds, pprehended from illegl trding nd kept in qurntine in CETAS/IBAMA, in the morning nd lte fternoon, from My to November It ws observed tht 1.82% of the smples collected in the morning were positive, while 31.36% of smples were positive in the lte fternoon. In ddition, the number of oocysts shed ws greter in the fternoon. Therefore, it ws concluded tht the smpling in the lte fternoon provided greter relibility for the dignosis of coccidiosis in green-winged-slttors. Moreover, in this study new isosporoid coccidin prsite from the green-winged-slttor S. similis ws observed nd is herein described. Isospor similisi n. sp. oocysts re spheroidl to sub-spheroidl, μm, with smooth nd bi-lyered wll, ~1.2 mm. Micropyle nd oocyst residuum re bsent, but splinter-like or comm-like grnules re present. Sporocysts re ellipsoidl or slightly ovoidl, mm. A stied body nd substied body re present. The sporocyst residuum is composed of grnules of different sizes. Sporozoites re vermiform with single refrctile body nd nucleus. This is the fourth description of n isosporoid coccidium infecting S. similis nd the sixth description from Crdinlide. Keywords: Periodicity, coccidin, oocysts, morphology, dignosis. Resumo A periodicidde diurn é um fenômeno que tem sido observdo em coccídios do gênero Isospor prsits de pássros, os quis eliminm um mior quntidde de oocistos o entrdecer. O objetivo deste estudo foi determinr periodicidde de eliminção de oocistos pels fezes no trinc-ferro-verddeiro Slttor similis, e su utilizção no dignóstico d coccidiose. Form colhids 220 mostrs fecis de ves oriunds de preensões do tráfico de nimis silvestres e mntids n qurenten do CETAS/IBAMA, nos períodos d mnhã e o entrdecer, de mio novembro de Observou-se que 1,82% ds mostrs colhids no período d mnhã form positivs, enqunto que 31,36% ds mostrs colhids form positivs o entrdecer, onde o mior número de oocistos foi observdo no período d trde. Portnto, concluiu-se que colheit de mostrs o entrdecer oferece mior confibilidde pr o dignóstico d coccidiose. Além disso, descreve-se um novo coccídio do trinc-ferro-verddeiro S. similis. Isospor similisi n. sp. possui oocistos esféricos subesféricos, 27,5 25,9 μm, com prede dupl e lis, ~1,2 µm. A micrópil e o resíduo do oocisto estão usentes, porém pequenos grânulos estão presentes. Os esporocistos são elipsóides ou levemente ovóides, 17,4 12,2 µm. Os corpos de Stied e substied estão presentes. O resíduo do esporocisto está presente e os esporozoítos possuem um corpo refrátil posterior e um núcleo. Est é qurt espécie isosporóide descrit de S. similis e sext descrição n fmíli Crdinlide. Plvrs-chve: Periodicidde, coccídios, oocistos, morfologi, dignóstico. *Corresponding uthor: Crlos Wilson Gomes Lopes Deprtmento de Prsitologi Animl, Instituto de Veterinári, Universidde Federl Rurl do Rio de Jneiro UFRRJ, BR-465, Km 7, CEP , Seropédic, RJ, Brsil e-mil: lopescwg@ufrrj.br

2 v. 22, n. 1, jn.-mr Oocyst shedding by Slttor similis nd Isospor similisi sp. n. 65 Introduction The green-winged slttor Slttor similis Lfresnye d Orbigny, 1837 is crdinlid bird resident in Brzil. Its nturl popultions hve been decimted by illegl mining nd trde, becuse of its beuty nd vocl repertoire. Among the informtion vilble on this species, it is known tht it does not present sexul dimorphism nd its locomotion is minly by jumping (SICK, 1997, IUCN, 2011). Estimtion of the intensity of coccidil infection is essentil for studies of the impct of prsitism upon nturl popultions (DOLNIK, 2006). The prsite-host reltionship in the species of birds, which hs been the focus of reserch over the pst decde, hs gret relevnce to coccidin intestinl prsites, which prsitize the mjority of vertebrte species nd re closely ssocited with the ecology of birds (LÓPEZ et l., 2007). The phenomenon of diurnl periodicity hs been observed in species of intestinl prsites, especilly the genus Isospor Schneider, 1881 (Protozo: Apicomplex) in wild birds, which eliminted their oocysts more frequently in the lte fternoon (BOUGHTON, 1937; DOLNIK, 1999, 2006; BROWN et l., 2001; LÓPEZ et l., 2007; MARTINAUD et l., 2009). Mny studies on the periodicity of elimintion of oocysts hve been conducted in severl species of psserines such s: blckcp Sylvi tricpill L., 1758; islnd cnry Serinus cnri L., 1758; grden wrbler Sylvi borin Boddert, 1783; Eursin blckbird Turdus merul L., 1758; Europen greenfinch Crduelis chloris L., 1758; house finch Crpodcus mexicnus Müller, 1776; smll ground-finch Geospiz fuliginos Gould, 1837; regent honeyeter Xnthomyz phrygi Shw, 1794; nd house sprrow Psser domesticus L., 1758 (BOX, 1977; BRAWNER III; HILL, 1999; BROWN et l., 2001; DOLNIK, 2006; LÓPEZ et l., 2007; LINDSTROM et l., 2009; MARTINAUD et l., 2009; DOLNIK et l., 2010; MORIN- ADELINE et l., 2011; PAP et l., 2011). However, in species of Brzilin psserines, there re no reports regrding diurnl periodicity in studies of prevlence or for dignosis of coccidiosis in free-living wild birds or birds kept in cptivity. Silv et l. (2010) reported tht during periods of reproduction nd moulting, wild birds kept in cptivity were more prone to infection by Isospor spp. This study imed to determine the periodicity of oocysts shedding nd identify new species of Isospor, not yet described in the scientific literture, from green-winged slttors S. similis recovered from the illegl trde of wild nimls, which were kept under qurntine in preprtion for relese. Mterils nd Methods In totl of 220 fecl smples were obtined from 164 greenwinged slttors S. similis recovered from the trfficking of wild nimls nd kept in qurntine in CETAS (Centro de Trigem de Animis Silvestres - Center for Trige of Wild Animls)/IBAMA (Instituto Brsileiro do Meio Ambiente e dos Recursos Nturis Renováveis - Brzilin Institute of Environment nd Nturl Resources)/MMA (Ministério do Meio Ambiente - Ministry of Environment) ( S nd W), which is locted t the Municiplity of Seropédic in the Stte of Rio de Jneiro, Brzil. The collection period ws from My to November Two hundred nd twenty smples were collected ccording to Dolnik (2006) nd Dolnik et l. (2009); fresh smples of feces, shed by bird, were collected individully from sheet of pper towel plced on the bottom of ech cge, fter clening in the morning (9-12h) nd t lte fternoon (15-17h). The collected smples were plced into plstic vils contining 2.5% (w/v) solution of K 2 Cr 2 O 7 t rtio of 1:6 (v/v). Smples were sent to the Coccidi nd Coccidiosis Lbortory t Universidde Federl Rurl do Rio de Jneiro (UFRRJ) for nlysis. Upon receipt they were plced in thin lyer (~5 mm) of K 2 Cr 2 O 7 2.5% solution in Petri pltes, with incubtion t C for 10 dys or until 70% of oocysts hd sporulted. Oocysts were recovered by flottion in Shether s sugr solution (S.G. 1.20) nd were exmined microscopiclly using the technique described by Duszynski nd Wilber (1997). The quntifiction of oocysts ws conducted ccording by Dolnik (2006) nd Dolnik et l. (2010), where the whole surfce of the coverslip ws observed to void counting errors which my be cused by the gglomertion of oocysts in some unobserved field. The results were expressed s oocysts per droplet (OoPD). An dditionl 108 smples were used to determine ech the periodicity of shedding of ech Isospor species. 54 smples were collected in the morning, between 9:00-12:00 PM, nd 54 smples ws collected in the lte fternoon (3:00-5:00 PM). In order to describe the fourth (novel) species, sporulted oocysts were obtined from smples collected in lte fternoon. Morphologicl observtions (micropyle [M], oocyst residuum [OR], polr grnule [PG], Stied body [SB], substied body [SSB], prstied body [PSB], sporocyst residuum [SR], sporozoite [SZ], refrctile body [SRB], nucleus [N]) nd mesurements, in µm, were performed using binoculr microscope Crl Zeiss with pochromtic oil immersion objective lens nd oculr micrometer (K-15X PZO, Polnd). Line drwings were prepred using binoculr microscope Wild M-20 with drwing tube. Pictures were tken using digitl cmer (Model CD Mvic MVC-CD250 Sony ). Size rnges re shown in prenthesis followed by verge nd shpe index (L/W rtio). Sttisticl nlysis by Fisher s test nd Wilcoxon s test were performed using Excel XP (Microsoft Co., Redmond, WA, USA), s proposed by Smpio (2002). Results Seventy-three of 220 smples exmined in this study for evlution of the incidence of oocysts shedding, were positive for Isospor spp. Only 1.82% of the smples collected in the morning hd positive results. In the lte fternoon, 31.36% of the smples were positive (Tble 1). Furthermore, oocyst shedding ws quntittively greter in the lte fternoon, with men vlue of 98 OoPD, in comprison with the men vlue of <1 OoPD recorded for the morning period (Tble 2). These results were highly significnt (p = ). All of the sporulted oocysts exmined were typicl of the genus Isospor. Four morphotypes of oocysts were recorded. The first three

3 66 Coelho, C.D. et l. Rev. Brs. Prsitol. Vet. Tble 1. Fecl smples of green-winged slttors, Slttor similis, used in the evlution of the incidence of oocyst shedding. Period Presence of oocysts Totl P vlue b b Morning Positive 4 ( 1.82 %) 110 (50%) Negtive 106 (48.18 %) Lte fternoon Positive 69 (31.36 %) 110 (50%) Negtive 41 (18.64 %) c Totl 220 (100%) Smpling in the period of the morning, between 9:00-12:00 AM, nd lte fternoon, between 3:00-5:00 PM. b Highly significnt by Fisher s test. Tble 2. Distribution of the men vlues of number of oocysts in the fecl smples of green-winged slttors, Slttor similis, by period. Period OoPD b P vlue c Morning N Men vlue 0.04 ( ) Lte fternoon N 110 Men vlue 98.3 ( ) Smpling in the period of the morning, between 9:00-12:00 AM, nd lte fternoon, between 3:00-5:00 PM. b Number of oocysts per droplet. c Highly significnt by Wilcoxon s test. types were identified s: (1) Isospor vnriperorum Levine, 1982; (2) Isospor slttori Berto et l., 2008; (3) nd Isospor trincferri Berto et l., The fourth type of oocysts ws different from the bove cited species nd ws considered to represent n unknown species, the description of which is provided below. Isospor similisi n. sp. (Figures 1-c, 2, b) Description of sporulted oocyst: Oocyst shpe (N = 25): spheroidl to sub-spheroidl; number of wlls: 2; wll thickness: 1.2 ( ); smooth outer wll bout 2/3 of totl thickness; L x W: ( ), with L/W rtio: 1.1 ( ); M nd OR: bsent; PG: present, forming splinter-like or comm-like grnules. Description of sporocyst nd sporozoites: Sporocyst shpe (N = 25): ellipsoidl or slightly ovoidl; L x W: ( ); L/W rtio: 1.4 ( ); SB: present, knoblike, 1.4 high 2.6 wide; SSB: present, lrge, homogeneous, 2.6 high 4.5 wide; PSB: bsent; SR: present; SR chrcteristics: composed of grnules of different sizes; SZ: vermiform with 1 posterior SRB nd centrlly locted N. Type host: Slttor similis Lfresnye d Orbigny, 1837 (Aves: Psseriformes: Crdinlide). Type mteril: One-hlf of the oocysts re kept in 10% queous buffered formlin (v/v) nd the other hlf in 70% ethnol ccording Duszynski nd Grdner (1991). Both smples were deposited in the Prsitology Collection, in the Deprtment of Animl Prsitology, t UFRRJ, locted in Seropédic, Rio de Figure 1. Line drwings of Isospor similisi n. sp, new coccidium species recovered from the green-winged slttor Slttor similis. () Sporulted oocyst with its respective vritions of (b, c) detched Stied nd substied bodies. Scle-br: 10 µm for oocysts; nd 5 µm for Stied nd substied bodies. Jneiro, Brzil. Phototypes nd line drwings re deposited s well. The repository number is P-41/2011. Type loclity: Seropédic. ( S nd W), Rio de Jneiro, Brzil. Sporultion time: seven dys. Site of infection: Not investigted. Prevlence: 59% (32 of 54 birds exmined). Etymology: The specific epithet is derived from the specific nme of the type host. Comments: Isospor similisi differs from other Isospor spp. from the sme host fmily (Tble 3). Only I. trincferri hd similr dimensions with I. similisi; however, it cn be distinguished by the unique PG nd bubble-shped SB (BERTO et l., 2008, 2011b). The periodicity ssocited with Isospor spp. is presented in the Tble 4. Firstly, lrger number of oocysts were recovered in the lte fternoon. Secondly, in the lte fternoon, four Isospor spp. were indentified (I. trincferri; I. vnriperorum; I. slttori; nd I. similisi), in contrst, in the morning only I. trincferri nd I. vnriperorum were indentified. Isospor similisi showed the highest men of OoPD in the lte fternoon; nd I. trincferri nd I. vnriperorum hve equl men vlues of OoPD in the morning. Discussion This study determined the most probble period of oocyst shedding, which should result in more relible dignosis of coccidin infection during qurntine t center for wild nimls, where the psserines hd suffered constnt stress since their cpture until their seizure by IBAMA. During winter in Rio de Jneiro, Brzil, the sunset hppens erlier, between 5:00-6:00 PM nd the smples used were collected in utumn, winter nd spring. The period of smpling occurred in the morning, between 9:00 12:00 PM, nd in lte fternoon, between 3:00-5:00 PM, which covered the min times for tretment nd mngement of niml sheld t CETAS. According to Brwner III nd Hill (1999), nd Villnú et l.

4 v. 22, n. 1, jn.-mr Oocyst shedding by Slttor similis nd Isospor similisi sp. n. b Figure 2. Photogrphs (-b) of sporulted oocysts of Isospor similisi n. sp, new coccidium species recovered from the green-winged slttor Slttor similis. Scle-br: 10 µm. (2006), the time of dy is crucil fctor tht must be tken into considertion when undertking the smpling of bird droppings. The mjority of species of coccidi tht infect psserines belong to the genus Isospor nd during their developmentl cycle, which includes oocyst shedding, there is diurnl periodicity (BOUGHTON, 1937; DOLNIK, 2006). In the present study only 1.82% of the smples collected in the morning were positive for coccidi but in the lte fternoon, 31.36% of the smples were positive. Thus, these results re similr to those of Boughton (1933, 1937, 1988), Dolnik (1999, 2006), Brown et l. (2001), López et l. (2007), Lindstrom et l. (2009), Mrtinud et l. (2009), Filipik et l. (2009), Dolnik et l. (2010), Morin-Adeline et l. (2011) nd Pp et l. (2011), where, in spite of different ltitudes nd longitudes, it ws reported tht the lte fternoon ws the most relible period for smpling becuse it is the period during which oocyst shedding is gretest. In birds kept in cptivity, the oocysts shedding begin erlier nd ended lter, when compred with the free-living psserines which hve pek of oocysts shedding between 1:00-9:00 PM. Moreover, the number of oocysts shed during the first dys of cptivity ws reported to be significntly higher thn during longer times post cpture (DOLNIK, 1999). Bsed on tht report, the smpling in the current study ws performed, whenever possible, on the first dy of rrivl of psserines in qurntine, with the objective of ensuring more relible dignosis. In study with house sprrows P. domesticus ws reported tht the periodicity of oocysts shedding ws not ssocited with feeding; however, it ws ssocited with photoperiod, becuse fter the reversl of light nd drkness, there is reversl in the time of oocyst shedding. On other hnd, it is known tht oocyst shedding is controlled by physiology of the host nd physiologicl differences between dietry hbits re probbly responsible for these differences (LÓPEZ et l., 2007). In the wild, birds show two peks of feeding ctivity, one in the morning nd nother in the lte fternoon. During this period, the birds feed in the sme res. In the breeding seson, they feed on the sme territory ll dy nd, during the migrtion seson, mny species gther in flocks to feed. Clerly, the presence of prsites in feces during this period would increse the chnces of infecting new hosts (DOLNIK, 1999; MARTINAUD et l., 2009). It is of vlue to note, tht, lthough the oocysts re reltively resistnt to environmentl fctors, such s temperture nd reltive humidity, it hs been reported tht desicction cn reduce the infectivity of oocysts of Eimeri spp. in poultry production; this lck of infectivity ws confirmed in n experiment with Isospor infection in Eursin blckbirds T. merul. It ws observed tht the relese of oocysts in the lte fternoon represented n dpttion to provide resistnce to desicction nd ultrviolet rdition, becuse if oocysts were relesed throughout the dy, most would be quickly destroyed by the ction of sunlight (MARTINAUD et l., 2009). In some species of birds which begin to migrte, the feeding pek disppers in the morning, but the feeding pek t lte fternoon never disppers. Thus, these birds gther with others, t lest in period of dy, incresing the chnces of infection of new hosts (DOLNIK, 1999). Finlly, we conclude tht estimtions of the prevlence nd prsite lod performed without considering the time of dy re not relible. A good method for ccurcy would be to restrict the period of smpling. Collection of smples for quntifiction of prsite lod for coccidi would be restricted to the second hlf of the lte fternoon, while those intended for prevlence studies could be crried out between 1/2 nd 4/5 of the lte fternoon s suggested by López et l. (2007). The results of the current study were similr, becuse the smpling ws restricted to the lte fternoon, specificlly between 3:00-5:00 PM. The oocysts shedding ws quntittively greter in the lte fternoon, with men vlue of 98 OoPD, in comprison with the men vlue of <1 OoPD in the morning period, even in wild psserines kept in qurntine regime. The illegl niml trde hd led to the reduction nd extinction of some bird species worldwide. In ddition, nother consequence is the introduction of new prsites, s occurred with Isospor vnriperorum Levine, 1982, first described from the northern

5 68 Coelho, C.D. et l. Rev. Brs. Prsitol. Vet. Tble 3. Comprtive morphology of Isospor similisi n. sp nd Isospor spp. recorded from crdinlid birds. Oocysts Sporocysts Coccidi Host(s) Reference(s)s Shpe Shpe index Wll (µm) Shpe Isospor vnriperorum Levine, 1982 Isospor pityli McQuistion & Cpprell, 1992 Isospor formrum McQuistion & Cpprell, 1992 Isospor slttori Berto, Blthzr, Flusino & Lopes, 2008 Crdinlis crdinlis Linneus, 1758; Slttor similis Lfresnye d Orbigny, 1837 Slttor grossus sturtus Todd, 1922 Slttor grossus grossus Linneus, 1766; S. g. sturtus Levine (1980) nd Levine et l. (1982) Lopes et l. (2007) McQuistion nd Cpprell (1992) McQuistion nd Cpprell (1992) sub-sphericl sub-sphericl sub-sphericl S. similis Berto et l. (2008) sub-sphericl Mesurements (µm) ( ) ( ) ( ) ( ) ( ) 1.0 ( ) 1.0 ( ) onelyered, c.0.8 c.1.5 c.1.5 c.1.1 Polr grnule present ovoid bsent ovoid bsent ovoid bsent ovoid Mesurements (µm) ( ) ( ) ( ) ( ) Stied body Substied body Residuum knob-like indistinct compct nipplelike smll, flttened bsent compct lrge, tringulr or conicl compct smll compct Isospor trincferri Berto, Blthzr, Flusino & Lopes, 2008 S. similis Berto et l. (2008) sub-sphericl ( ) 1.1 ( ) c.1.2 present ovoid ( ) smll, nipplelike bubbleshped prominent diffuse Isospor similisi n. sp. S. similis current study sphericl to sub-sphericl ( ) 1.1 ( ) c.1.2 present, splinterlike or comm-like grnules ellipsoidl or slightly ovoidl ( ) knob-like lrge, homogeneous diffuse

6 v. 22, n. 1, jn.-mr Oocyst shedding by Slttor similis nd Isospor similisi sp. n. 69 Tble 4. Periodicity of shedding Isospor oocysts from green-winged slttors Slttor similis t CETAS/IBAMA, Seropédic in the Stte of Rio de Jneiro, Brzil. Species Morning (n = 54) Lte fternoon b (n = 54) OoPD c Mens OoPD c Mens I. trincferri I. vnriperorum I. slttori I. similisi Totl Smpling in the period of the morning, between 9:00-12:00 AM. b Smpling in the lte fternoon, between 3:00-5:00 PM. c Number of oocysts per droplet. crdinl Crdinlis crdinlis L., 1758, but which due to its introduction into South Americ for breeding in cptivity, ws subsequently reported from S. similis (LOPES et l., 2007; BERTO et l., 2011b). The ese of trnsmission of Isospor mong birds from the sme fmily, the lrge number of crdinlid species in New World nd the illegl bird trde my serve to ensure wide distribution of these prsites in South, Centrl nd North Americ (BERTO et l., 2011b). In Brzil, the ntive birds re protected by lw nd supervised by the Brzilin Institute of the Environment nd Nturl Renewble Resources (IBAMA), therefore symbiotype could not be collected (IBAMA, 2011). According to Duszynski nd Wilber (1997) nd Berto et l. (2011b), new coccidin species should be morphologiclly compred with other congeneric coccidin species, tht shre similr fetures nd belong to the sme host fmily. Thus, I. similisi sporulted oocysts ws compred to ll of described species from Crdinlide. As observed in the current study, few vritions in SB nd SSB (Figure 1b, c) were lso observed in sporocysts of I. similisi ccording to Grulet et l. (1982), Berto et l. (2009, b, c, 2010, 2011, c), Coelho et l. (2011, b) nd Pereir et l. (2011); however, these vritions re not significnt enough to seprte nd describe it s new species. These vritions could possibly be the result of the sporultion process, the position of the SZ inside of the sporocyst, or the position of the oocyst nd sporocyst under the coverslip. Bsed upon its morphologicl fetures, I. similisi is considered s new to science, being the fourth description from S. similis nd the sixth description from the host fmily Crdinlide. Interestingly, bsed upon the OoPD vlues, this species ws considered to be the most competitive mong the four species of Isospor ssocited with S. similis. Acknowledgements We re thnkful to stff t the CETAS/IBAMA/MMA fcility t the Municiplity of Seropédic in the Stte of Rio de Jneiro, who enbled the collection of smples from birds held for rehbilittion nd reintroduction into the wild s well s CNPq nd FAPERJ. References Berto BP, Blthzr LMC, Flusino W, Lopes CWG. Two new coccidin prsites of green-winged slttor (Slttor similis) from South Americ. Act Protozool 2008; 47(3): Berto BP, Flusino W, Luz HR, Ferreir I, Lopes CWG. Isospor coerebe n. sp. (Apicomplex: Eimeriide) from the bnnquit Coereb flveol (Psseriformes: Coerebide) in South Americ. Rev Brs Prsitol Vet 2011; 20(1): PMid: S Berto BP, Flusino W, Luz HR, Ferreir I, Lopes CWG. Isospor mionectesi sp. nov. (Apicomplex, Eimeriide) from the grey-hooded flyctcher, Mionectes rufiventris in Brzil. Act Prsitol 2009; 54(4): Berto BP, Flusino W, Luz HR, Ferreir I, Lopes CWG. Isospor rmphoceli n. sp. from the Brzilin tnger (Rmphocelus bresilius dorslis) in South Americ. Zootx 2010; 2650(1): Berto BP, Flusino W, McIntosh D, Teixeir-Filho WL, Lopes CWG. Coccidi of New World psserine birds (Aves: Psseriformes): review of Eimeri Schneider, 1875 nd Isospor Schneider, 1881 (Apicomplex: Eimeriide). Syst Prsitol 2011b; 80(3): PMid: Berto BP, Luz HR, Flusino W, Ferreir I, Lopes CWG. New species of Eimeri Schneider, 1875 nd Isospor Schneider, 1881 (Apicomplex: Eimeriide) from the short-crested flyctcher Myirchus ferox (Gmelin) (Psseriformes: Tyrnnide) in South Americ. 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