Parasitol Res (2014) 113:3953–3955 DOI 10.1007/s00436-014-4059-6
ORIGINAL PAPER
Isospora dromaii n. sp. (Apicomplexa, Eimeriidae) isolated from emus, Dromaius novaehollandiae (Casuariiformes, Casuariidae) Carina dos Santos Teixeira & Samira Salim Mello Gallo & Nicole Brand Ederli & Bruno Pereira Berto & Francisco Carlos Rodrigues de Oliveira
Received: 9 April 2014 / Accepted: 23 July 2014 / Published online: 7 September 2014 # Springer-Verlag Berlin Heidelberg 2014
Abstract A new species of Coccidia (Protozoa: Apicomplexa: Eimeriidae) isolated from emus, Dromaius novaehollandiae, which was observed in Brazil is described and named. Oocysts of Isospora dromaii n. sp. are subspheroidal to ovoid in shape, measure 21.6×19.8 μm, and have a double and smooth wall thickness of approximately 1.4 μm. In this species, micropyle, oocyst residuum, and polar granules are absent. The sporocysts are slightly ovoid in shape and measure 13.7×10.0 μm. Nipple-like Stieda body and prominent sub-Stieda body are present. The sporocyst residuum is composed of small dispersed granules of varying sizes. The sporozoites are characterized by an oblong refractile body and one centrally located nucleus. This is the first description of isosporid coccidia infecting birds of the family Dromaiidae.
Keywords Isospora . Eimeriidae . Coccidia . Oocyst . Ratites
C. dos Santos Teixeira : S. S. M. Gallo : F. C. R. de Oliveira (*) Laboratório de Sanidade Animal, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, Rio de Janeiro 28035-602, Brazil e-mail: [email protected] N. B. Ederli Universidade Federal do Pará, Campus Universitário do Marajó–Breves, Breves, Pará 68800-000, Brazil B. P. Berto Departamento de Biologia Animal, Instituto de Biologia, Universidade Federal Rural do Rio de Janeiro, Seropédica Rio de Janeiro 23897-970, Brazil
Introduction The emu, Dromaius novaehollandiae (Latham, 1790), is a large flightless bird in the ratite group with the ostrich, Struthio camelus Linnaeus, 1758; the rhea, Rhea americana Linnaeus, 1758 and Rhea pennata (Orbigny, 1834); the cassowary, Casuarius casuarius (Linnaeus, 1758) (displaying a closer relationship); and the kiwi, Apteryx spp. (Shaw, 1813) (Jeffrey 2001). Taxonomically, the emu is classified in the order Casuariiformes (Forbes 1884), family Casuariidae Kaup, 1847, and genus Dromaius Vieillot, 1816 (Christidis and Boles 2008). Emus are native to Australia, occupying most of the continent except tropical forests and cleared agricultural areas (Davies 1976). Since the mid-1980s, there has been growing interest in the captive breeding of emus for the production of leather, meat, and oil (Malecki et al. 1998). Coccidiosis in poultry is caused by protozoa of the genus Eimeria or Isospora. Each species displays specific characteristics regarding prevalence, site of infection, pathogenicity, and immunogenicity (Rose and Long 1980). However, all species parasitize intestinal epithelial cells causing pathological alterations of the mucosa and adjacent tissue. These lesions can cause serious bleeding and systemic effects, such as shock syndrome and death. Nevertheless, in many cases, immunity develops after repeated infection. These lesions can cause decreased egg production and malabsorption (less weight gain and lower feed conversion), even in the absence of clinical signs, thereby resulting in economic loss (Vermeulen et al. 2001). A few incidents of coccidiosis in emus have been reported. Recently, Papini et al. (2012) reported non-sporulated oocysts in the feces of two emus, D. novaehollandiae, from a zoo in Italy. However, these oocysts were not described, measured, or named.
3954
The present study describes a new species of coccidia infecting D. novaehollandiae raised in the city of Campos dos Goytacazes, State of Rio de Janeiro, Brazil.
Material and methods Fresh fecal samples were collected from 13 birds from a scientific breeding colony at the Universidade Estadual do Norte Fluminense Darcy Ribeiro, located in the city of Campos dos Goytacazes, State of Rio de Janeiro, Brazil. The samples were collected in individual bags that were labeled and packed in ice boxes (8–10 °C) and were immediately transported to the laboratory located at the university. The feces were mixed with a 2.5 % potassium dichromate solution (K2Cr2O7), filtered with double gauze, and aerated with the aid of an aquarium pump coupled to plastic tubes to facilitate sporulation. Oocysts were recovered via Sheather’s flotation technique with sucrose solution with a density of 1.3 g/ml and were examined microscopically using the technique described by Duszynski and Wilber (1997). Images were captured using a digital camera (Canon PowerShot A640, USA) coupled to a binocular microscope (Carl Zeiss, Germany). Measurements were made using a Zeiss AxioVision Sample Images Software and are provided in micrometers. Drawings were generated by overlapping images in different focuses with the aid of the software Corel Photo Paint version 11. The size ranges are provided in parentheses, followed by the mean, standard deviation, and morphometric ratio (length/width). Description Eleven (nine females and two males) of 13 emus examined were shedding oocysts in the feces during the reproductive period. Oocyst sporulation started at the 15th day after aeration, and at the 20th day, 70 % of oocysts were sporulated. Higher sporulation rates up to 25 days were not observed, which was the maximum period of observation. All oocysts observed were characteristic of Isospora, which is described and named as follows:
Parasitol Res (2014) 113:3953–3955
Sporulated oocysts Oocyst shape (n=62): subspheroidal to ovoid; double-layered wall, approximately 1.4±0.3 (0.8 to 1.9) thick; smooth outer wall representing two thirds of the total thickness; L×W=21.6 ±2.9 (12.3 to 28.3)×19.8±2.7 (11.8 to 25.9), L/W=1.1±0.1 (0.9 to 1.3); micropyle, oocyst residuum, and polar granules absent. Sporocysts and sporozoites Sporocysts shape (n=54): slightly ovoid; L×W=13.7±2.1 (9.2 to 18.9)×10.0±1.2 (7.5 to 13.5); L/W=1.4±0.2 (1.0 to 1.7); Stieda body present, nipple-like in shape, L×W=1.7±0.2 (1.4 to 2.1)×1.0±0.3 (0.6 to 1.4); prominent sub-Stieda body present, L×W=4.7±0.9 (3.3 to 5.7)×1.9±1.0 (0.8 to 3.6); para-Stieda body absent; sporocyst residuum composed of dispersed small, compact spherical granules of different sizes; elongated sporozoites, with a clearly visible oblong refractile body and one nucleus centrally located.
Discussion There are few species of coccidia reported from Casuariidae birds and only a single species of Isospora described from ratite birds, S. camelus (Yakimoff 1940). In this study, oocysts of I. dromaii n. sp. isolated from emu feces displayed a subspheroidal to ovoid shape with a double wall. Meanwhile, oocysts derived from ostriches (Isospora struthionis), as reported by Yakimoff (1940), were described as rounded with a double wall. Isospora from emus do not present polar granules, unlike the oocysts found in ostriches, which present
Isospora dromaii n. sp. Host type: D. novaehollandiae (Aves: Casuariiformes: Casuariidae). Locality type: Scientific breeding at the Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, State of Rio de Janeiro, Brazil (21° 45′ 16″ S; 41° 19′ 28″ W). Prevalence = 11/13 (84.6 %). Sporulation = 15 to 25 days. Site of infection: Unknown. Etymology: The species epithet is derived from the host specific name.
Fig. 1 Schematic drawing of Isospora dromaii n. sp. a Sporulated oocysts, b variation in Stieda and sub-Stieda bodies. Bar = 10 μm
Parasitol Res (2014) 113:3953–3955
3955
Fig. 2 Optical micrograph of Isospora dromaii n. sp. sporulated oocyst (a, b). Bar = 10 μm
double polar granules. I. dromaii n. sp. sporocysts are ovoid, with a nipple-like Stieda body, a prominent sub-Stieda body, and a residuum composed of dispersed, small, spherical, and compact granules of varying sizes. In contrast, I. struthionis sporocysts present with a lemon shape and pointed Stieda body, while sub-Stieda bodies and residual granules are absent. I. dromaii sporozoites present with a clearly visible oblong refractile body and one nucleus, while I. struthionis sporozoites were not characterized. In our study, oocysts sporulated at 15 to 25 days (Figs. 1 and 2). However, the coccidian sporulation period of ratites has been poorly characterized, and Faust and Pappas (1977) have isolated oocysts from rheas, R. americana, from the Columbus Zoo in Ohio, USA, which sporulated after 2 days at room temperature. Although they were characterized as Isospora sp., they displayed a reduced sporulation time compared to those shed by emus in our study. The mean diameter of oocysts and sporocysts isolated from emus in the present study were smaller than the diameter measurements cited by Yakimoff (1940) from ostriches (e.g., oocysts approximately 30.6 μm and sporocysts approximately 18.0×10.8 μm). However, the oocysts in the feces derived from emus in our study displayed greater measurements than those isolated from rheas in Europe (12–15 μm), as described by Ponce Gordo et al. (2002). These researchers did not cite the species of the measured oocysts, most likely because the oocysts measured were non-sporing. Based on the variations in morphological and morphometric characteristics mentioned in our study, I. dromaii is
considered as a new species and the first isosporid coccidia found in D. novaehollandiae emus.
References Christidis L, Boles WE (2008) Systematics and taxonomy of Australian birds. CSIRO, Melbourne Davies SJJF (1976) The natural history of the emu in comparison with that of other ratites. In: Frith HJ, Calaby JH (eds) The 16th international ornithological congress. Australian Academy of Science, Canberra, pp 109–120 Duszynski DW, Wilber PG (1997) A guideline for the preparation of species descriptions in the Eimeridae. J Parasitol 83:333–336 Faust BS, Pappas PW (1977) A survey of coccidia and helminth parasites of birds at the Columbus (Ohio) Zoo. J Zoo Anim Med 8:18–23 Jeffrey JS (2001) Emu production. http://gallus.tamu.edu/Extension% 20publications/emuproduction.pdf. Accessed 16 Oct 2013 Malecki IA, Martin GB, O’Malley PJ, Meyer GT, Talbot RT, Sharp PJ (1998) Endocrine and testicular changes in a short-day seasonally breeding bird, the emu (Dromaius novaehollandiae), in southwestern Australia. Anim Reprod Sci 53:143–155 Papini R, Girivetto M, Marangi M, Mancianti F, Giangaspero A (2012) Endoparasite infections in pet and zoo birds in Italy. Sci World J 2012:1–9 Ponce Gordo F, Herrera S, Castro AT, García Durán B, Martínez Díaz RA (2002) Parasites from ostriches (Strutio camelus) and rheas (Rhea americana) in Europe. Vet Parasit 107:137–160 Rose ME, Long PL (1980) Vaccination against coccidiosis in chickens. In: Taylor AER, Muller R (eds) Vaccination against parasites. Blackwell Scientific, Oxford, pp 57–74 Vermeulen AN, Schaap DC, Schetters TPM (2001) Control of coccidiosis in chickens by vaccination. Vet Parasitol 100:13–20 Yakimoff WL (1940) Isospora struthionis n. sp., coccidie de l’autruche africaine. Ann Soc Med Trop 20:137–138
ORIGINAL PAPER
Isospora dromaii n. sp. (Apicomplexa, Eimeriidae) isolated from emus, Dromaius novaehollandiae (Casuariiformes, Casuariidae) Carina dos Santos Teixeira & Samira Salim Mello Gallo & Nicole Brand Ederli & Bruno Pereira Berto & Francisco Carlos Rodrigues de Oliveira
Received: 9 April 2014 / Accepted: 23 July 2014 / Published online: 7 September 2014 # Springer-Verlag Berlin Heidelberg 2014
Abstract A new species of Coccidia (Protozoa: Apicomplexa: Eimeriidae) isolated from emus, Dromaius novaehollandiae, which was observed in Brazil is described and named. Oocysts of Isospora dromaii n. sp. are subspheroidal to ovoid in shape, measure 21.6×19.8 μm, and have a double and smooth wall thickness of approximately 1.4 μm. In this species, micropyle, oocyst residuum, and polar granules are absent. The sporocysts are slightly ovoid in shape and measure 13.7×10.0 μm. Nipple-like Stieda body and prominent sub-Stieda body are present. The sporocyst residuum is composed of small dispersed granules of varying sizes. The sporozoites are characterized by an oblong refractile body and one centrally located nucleus. This is the first description of isosporid coccidia infecting birds of the family Dromaiidae.
Keywords Isospora . Eimeriidae . Coccidia . Oocyst . Ratites
C. dos Santos Teixeira : S. S. M. Gallo : F. C. R. de Oliveira (*) Laboratório de Sanidade Animal, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, Rio de Janeiro 28035-602, Brazil e-mail: [email protected] N. B. Ederli Universidade Federal do Pará, Campus Universitário do Marajó–Breves, Breves, Pará 68800-000, Brazil B. P. Berto Departamento de Biologia Animal, Instituto de Biologia, Universidade Federal Rural do Rio de Janeiro, Seropédica Rio de Janeiro 23897-970, Brazil
Introduction The emu, Dromaius novaehollandiae (Latham, 1790), is a large flightless bird in the ratite group with the ostrich, Struthio camelus Linnaeus, 1758; the rhea, Rhea americana Linnaeus, 1758 and Rhea pennata (Orbigny, 1834); the cassowary, Casuarius casuarius (Linnaeus, 1758) (displaying a closer relationship); and the kiwi, Apteryx spp. (Shaw, 1813) (Jeffrey 2001). Taxonomically, the emu is classified in the order Casuariiformes (Forbes 1884), family Casuariidae Kaup, 1847, and genus Dromaius Vieillot, 1816 (Christidis and Boles 2008). Emus are native to Australia, occupying most of the continent except tropical forests and cleared agricultural areas (Davies 1976). Since the mid-1980s, there has been growing interest in the captive breeding of emus for the production of leather, meat, and oil (Malecki et al. 1998). Coccidiosis in poultry is caused by protozoa of the genus Eimeria or Isospora. Each species displays specific characteristics regarding prevalence, site of infection, pathogenicity, and immunogenicity (Rose and Long 1980). However, all species parasitize intestinal epithelial cells causing pathological alterations of the mucosa and adjacent tissue. These lesions can cause serious bleeding and systemic effects, such as shock syndrome and death. Nevertheless, in many cases, immunity develops after repeated infection. These lesions can cause decreased egg production and malabsorption (less weight gain and lower feed conversion), even in the absence of clinical signs, thereby resulting in economic loss (Vermeulen et al. 2001). A few incidents of coccidiosis in emus have been reported. Recently, Papini et al. (2012) reported non-sporulated oocysts in the feces of two emus, D. novaehollandiae, from a zoo in Italy. However, these oocysts were not described, measured, or named.
3954
The present study describes a new species of coccidia infecting D. novaehollandiae raised in the city of Campos dos Goytacazes, State of Rio de Janeiro, Brazil.
Material and methods Fresh fecal samples were collected from 13 birds from a scientific breeding colony at the Universidade Estadual do Norte Fluminense Darcy Ribeiro, located in the city of Campos dos Goytacazes, State of Rio de Janeiro, Brazil. The samples were collected in individual bags that were labeled and packed in ice boxes (8–10 °C) and were immediately transported to the laboratory located at the university. The feces were mixed with a 2.5 % potassium dichromate solution (K2Cr2O7), filtered with double gauze, and aerated with the aid of an aquarium pump coupled to plastic tubes to facilitate sporulation. Oocysts were recovered via Sheather’s flotation technique with sucrose solution with a density of 1.3 g/ml and were examined microscopically using the technique described by Duszynski and Wilber (1997). Images were captured using a digital camera (Canon PowerShot A640, USA) coupled to a binocular microscope (Carl Zeiss, Germany). Measurements were made using a Zeiss AxioVision Sample Images Software and are provided in micrometers. Drawings were generated by overlapping images in different focuses with the aid of the software Corel Photo Paint version 11. The size ranges are provided in parentheses, followed by the mean, standard deviation, and morphometric ratio (length/width). Description Eleven (nine females and two males) of 13 emus examined were shedding oocysts in the feces during the reproductive period. Oocyst sporulation started at the 15th day after aeration, and at the 20th day, 70 % of oocysts were sporulated. Higher sporulation rates up to 25 days were not observed, which was the maximum period of observation. All oocysts observed were characteristic of Isospora, which is described and named as follows:
Parasitol Res (2014) 113:3953–3955
Sporulated oocysts Oocyst shape (n=62): subspheroidal to ovoid; double-layered wall, approximately 1.4±0.3 (0.8 to 1.9) thick; smooth outer wall representing two thirds of the total thickness; L×W=21.6 ±2.9 (12.3 to 28.3)×19.8±2.7 (11.8 to 25.9), L/W=1.1±0.1 (0.9 to 1.3); micropyle, oocyst residuum, and polar granules absent. Sporocysts and sporozoites Sporocysts shape (n=54): slightly ovoid; L×W=13.7±2.1 (9.2 to 18.9)×10.0±1.2 (7.5 to 13.5); L/W=1.4±0.2 (1.0 to 1.7); Stieda body present, nipple-like in shape, L×W=1.7±0.2 (1.4 to 2.1)×1.0±0.3 (0.6 to 1.4); prominent sub-Stieda body present, L×W=4.7±0.9 (3.3 to 5.7)×1.9±1.0 (0.8 to 3.6); para-Stieda body absent; sporocyst residuum composed of dispersed small, compact spherical granules of different sizes; elongated sporozoites, with a clearly visible oblong refractile body and one nucleus centrally located.
Discussion There are few species of coccidia reported from Casuariidae birds and only a single species of Isospora described from ratite birds, S. camelus (Yakimoff 1940). In this study, oocysts of I. dromaii n. sp. isolated from emu feces displayed a subspheroidal to ovoid shape with a double wall. Meanwhile, oocysts derived from ostriches (Isospora struthionis), as reported by Yakimoff (1940), were described as rounded with a double wall. Isospora from emus do not present polar granules, unlike the oocysts found in ostriches, which present
Isospora dromaii n. sp. Host type: D. novaehollandiae (Aves: Casuariiformes: Casuariidae). Locality type: Scientific breeding at the Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, State of Rio de Janeiro, Brazil (21° 45′ 16″ S; 41° 19′ 28″ W). Prevalence = 11/13 (84.6 %). Sporulation = 15 to 25 days. Site of infection: Unknown. Etymology: The species epithet is derived from the host specific name.
Fig. 1 Schematic drawing of Isospora dromaii n. sp. a Sporulated oocysts, b variation in Stieda and sub-Stieda bodies. Bar = 10 μm
Parasitol Res (2014) 113:3953–3955
3955
Fig. 2 Optical micrograph of Isospora dromaii n. sp. sporulated oocyst (a, b). Bar = 10 μm
double polar granules. I. dromaii n. sp. sporocysts are ovoid, with a nipple-like Stieda body, a prominent sub-Stieda body, and a residuum composed of dispersed, small, spherical, and compact granules of varying sizes. In contrast, I. struthionis sporocysts present with a lemon shape and pointed Stieda body, while sub-Stieda bodies and residual granules are absent. I. dromaii sporozoites present with a clearly visible oblong refractile body and one nucleus, while I. struthionis sporozoites were not characterized. In our study, oocysts sporulated at 15 to 25 days (Figs. 1 and 2). However, the coccidian sporulation period of ratites has been poorly characterized, and Faust and Pappas (1977) have isolated oocysts from rheas, R. americana, from the Columbus Zoo in Ohio, USA, which sporulated after 2 days at room temperature. Although they were characterized as Isospora sp., they displayed a reduced sporulation time compared to those shed by emus in our study. The mean diameter of oocysts and sporocysts isolated from emus in the present study were smaller than the diameter measurements cited by Yakimoff (1940) from ostriches (e.g., oocysts approximately 30.6 μm and sporocysts approximately 18.0×10.8 μm). However, the oocysts in the feces derived from emus in our study displayed greater measurements than those isolated from rheas in Europe (12–15 μm), as described by Ponce Gordo et al. (2002). These researchers did not cite the species of the measured oocysts, most likely because the oocysts measured were non-sporing. Based on the variations in morphological and morphometric characteristics mentioned in our study, I. dromaii is
considered as a new species and the first isosporid coccidia found in D. novaehollandiae emus.
References Christidis L, Boles WE (2008) Systematics and taxonomy of Australian birds. CSIRO, Melbourne Davies SJJF (1976) The natural history of the emu in comparison with that of other ratites. In: Frith HJ, Calaby JH (eds) The 16th international ornithological congress. Australian Academy of Science, Canberra, pp 109–120 Duszynski DW, Wilber PG (1997) A guideline for the preparation of species descriptions in the Eimeridae. J Parasitol 83:333–336 Faust BS, Pappas PW (1977) A survey of coccidia and helminth parasites of birds at the Columbus (Ohio) Zoo. J Zoo Anim Med 8:18–23 Jeffrey JS (2001) Emu production. http://gallus.tamu.edu/Extension% 20publications/emuproduction.pdf. Accessed 16 Oct 2013 Malecki IA, Martin GB, O’Malley PJ, Meyer GT, Talbot RT, Sharp PJ (1998) Endocrine and testicular changes in a short-day seasonally breeding bird, the emu (Dromaius novaehollandiae), in southwestern Australia. Anim Reprod Sci 53:143–155 Papini R, Girivetto M, Marangi M, Mancianti F, Giangaspero A (2012) Endoparasite infections in pet and zoo birds in Italy. Sci World J 2012:1–9 Ponce Gordo F, Herrera S, Castro AT, García Durán B, Martínez Díaz RA (2002) Parasites from ostriches (Strutio camelus) and rheas (Rhea americana) in Europe. Vet Parasit 107:137–160 Rose ME, Long PL (1980) Vaccination against coccidiosis in chickens. In: Taylor AER, Muller R (eds) Vaccination against parasites. Blackwell Scientific, Oxford, pp 57–74 Vermeulen AN, Schaap DC, Schetters TPM (2001) Control of coccidiosis in chickens by vaccination. Vet Parasitol 100:13–20 Yakimoff WL (1940) Isospora struthionis n. sp., coccidie de l’autruche africaine. Ann Soc Med Trop 20:137–138
