Artigo: Coccidial dispersion across New World marsupials- Klossiella tejerai 2014

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Systematic Parasitology
An International Journal
 
ISSN 0165-5752
Volume 89
Number 1
 
Syst Parasitol (2014) 89:83-89
DOI 10.1007/s11230-014-9510-7
Coccidial dispersion across New World
marsupials: Klossiella tejerai Scorza,
Torrealba & Dagert, 1957 (Apicomplexa:
Adeleorina) from the Brazilian common
opossum Didelphis aurita (Wied-Neuwied)
(Mammalia: Didelphimorphia)
Caroline Spitz dos Santos, Bruno Pereira
Berto, Bruno do Bomfim Lopes, et al.
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Coccidial dispersion across New World marsupials:
Klossiella tejerai Scorza, Torrealba & Dagert, 1957
(Apicomplexa: Adeleorina) from the Brazilian common
opossum Didelphis aurita (Wied-Neuwied) (Mammalia:
Didelphimorphia)
Caroline Spitz dos Santos • Bruno Pereira Berto • Bruno do Bomfim Lopes •
Matheus Dias Cordeiro • Adivaldo Henrique da Fonseca • Walter Leira Teixeira Filho •
Carlos Wilson Gomes Lopes
Received: 11 June 2014 / Accepted: 14 July 2014
� Springer Science+Business Media Dordrecht 2014
Abstract Klossiella tejerai Scorza, Torrealba &
Dagert, 1957 is a primitive coccidian parasite reported
from the New World marsupials Didelphis marsupialis
(Linnaeus) and Marmosa demerarae (Thomas). The
current work describes K. tejerai from the Brazilian
common opossum Didelphis aurita (Wied-Neuwied) in
Southeastern Brazil, evidencing the coccidial dispersion
across opossums of the same family. The sporocysts
recovered from urine samples were ellipsoidal,
20.4 9 12.7 lm, with sporocyst residuum composed
of scattered spherules and c.13 sporozoites per sporo-
cyst, with refractile bodies and nucleus. Macrogametes,
microgametes, sporonts, sporoblasts/sporocysts were
identified within parasitophorous vacuoles of epithelial
cells located near the renal corticomedullary junction.
Didelphis marsupialis should not have transmitted K.
tejerai to D. aurita because they are not sympatric;
however M. demerarae is sympatric with D. marsupialis
and D. aurita. Therefore, D. aurita becomes the third
host species for K. tejerai in South America.
Introduction
Opossums in the New World represent 99 different
species. The vast majority of these (95 species, 96%)
inhabits South America. Didelphis spp. are common in
South America; however, one of the six species,
Didelphis virginiana (Kerr) has distribution in North
and Central Americas (IUCN, 2014).
Didelphis spp. became epidemiologically relevant
in the New World when they were identified as
definitive hosts for some coccidian parasites of the
genus Sarcocystis Lankester, 1882. Among these
Sarcocystis spp., Sarcocystis neurona Dubey, Davis,
Speer, Bowman, Lahunta, Granstrom, Topper, Hamir,
Cummings & Suter, 1991 is recognised as the
C. S. dos Santos � M. D. Cordeiro
Curso de Po´s-Graduac¸a˜o em Cieˆncias Veterina´rias,
Universidade Federal Rural do Rio de Janeiro (UFRRJ),
BR-465 km 7, 23897-970 Serope´dica, RJ, Brazil
B. P. Berto (&)
Departamento de Biologia Animal, Instituto de Biologia,
UFRRJ, BR-465 km 7, 23897-970 Serope´dica, RJ, Brazil
e-mail: bertobp@ufrrj.br
B. do Bomfim Lopes
Programa de Po´s-graduac¸a˜o em Cieˆncia, Tecnologia e
Inovac¸a˜o em Agropecua´ria, UFRRJ, BR-465 km 7,
23897-970 Serope´dica, RJ, Brazil
A. H. da Fonseca
Departamento de Epidemiologia e Sau´de Pu´blica,
Instituto de Veterina´ria, UFRRJ, BR-465 km 7,
23897-970 Serope´dica, RJ, Brazil
W. L. T. Filho � C. W. G. Lopes
Departamento de Parasitologia Animal, Instituto de
Veterina´ria, UFRRJ, BR-465 km 7,
23897-970 Serope´dica, RJ, Brazil
123
Syst Parasitol (2014) 89:83–89
DOI 10.1007/s11230-014-9510-7
Author's personal copy
ethiological agent of equine protozoal myeloenceph-
alitis (Dubey & Lindsay, 1998; Monteiro et al., 2013).
Besides Sarcocystis spp., other coccidia infect
Didelphis spp., including Eimeria spp. (Teixeira
et al., 2007) and Klossiella tejerai Scorza, Torrealba
& Dagert, 1957 (see Scorza et al., 1957). Klossiella
spp. have been reported from various marsupials,
primarily from Australian peramelids, petaurids and
macropodids (Barker et al., 1975, 1985; Bennett et al.,
2007). However, in the New World, only K. tejerai
was described from Didelphis marsupialis (Linnaeus)
in Venezuela (Scorza et al., 1957), and subsequently
reported from this host species in Panama (Edgcomb
et al., 1976) and from the woolly mouse opossum
Marmosa demerarae (Thomas) in Guyana (Boulard,
1975).
The present study describes K. tejerai infecting a
Brazilian common opossum Didelphis aurita (Wied-
Neuwied) in Southeastern Brazil, evidencing the
coccidial dispersion across opossums of the same
family.
Materials and methods
Twenty opossums D. aurita were captured on and
around the Campus of the Federal Rural University of
Rio de Janeiro (Universidade Federal Rural do Rio de
Janeiro – UFRRJ), located in the municipality of
Serope´dica (22�440S, 43�420W), state of Rio de
Janeiro, Brazil. The opossums were transported to the
Veterinary Institute (Instituto de Veterina´ria – IV) at
the UFRRJ, and were reared and fed in small enclo-
sures approximately 1 9 1 m. Feed and water were
administered ad libitum. The capture, maintenance and
collection of samples was approved by UFRRJ Ethics
Committee under protocol No. 255/2012 and author-
ised by Brazilian Institute of Environment and Natural
Renewable Resources (Instituto Brasileiro do Meio
Ambiente e dos Recursos Naturais Renova´veis
– IBAMA) under protocol # 34701-2. Sample pro-
cessing and data analysis were conducted at the
Laboratory of Coccidia and Coccidiosis (Laborato´rio
de Coccı´dios e Coccidioses – LCC) located at UFRRJ.
Urine samples were collected and placed in plastic
vials. Sporocysts of Klossiella spp. were recovered by
centrifugal sedimentation and examined microscopi-
cally using the technique described by Duszynski &
Wilber (1997). The opossums positive for sporocysts
of Klossiella spp. in urine were necropsied. Kidneys
were examined grossly and representative samples of
kidney tissue were collected into 10% neutral buffered
formalin. Once fixed, these tissues were embedded in
paraffin, sectioned at 4 lm, and stained routinely with
hematoxylin and eosin. Morphological observations,
line drawing and photomicrographs were made using
an Olympus BX binocular microscope coupled to a
digital camera Eurocam 5.0. All measurements are in
micrometres and are presented as the range followed
by the mean.
Fig. 1 Urinary sporocysts of Klossiella tejerai from the
Brazilian common opossum Didelphis aurita. A, Composite
line drawing; B–C, Photomicrographs. Scale-bars: 10 lm
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Results
Twenty Brazilian common opossums were examined;
one of them (5%) shed Klossiella-like sporocysts in
the urine. The current description follows the guide-
lines of Duszynski & Wilber (1997) and Berto et al.
(2014a) for the urinary sporocysts and the examples of
Scorza et al. (1957), Barker etal. (1975; 1985),
Gardiner et al. (1998) and Bennett et al. (2007) for the
nomenclature of tissue stages of Klossiella.
Klossiella tejerai Scorza, Torrealba & Dagert, 1957
Host: Didelphis aurita Wied-Neuwied (Mammalia:
Didelphimorphia: Didelphidae).
Locality: Brazil, State of Rio de Janeiro, Municipality
of Serope´dica (22�440S, 43�420W).
Material studied: One-half of the sporocysts from
urine samples are kept in 10% aqueous buffered
formalin (v/v) and the other half in 70% ethanol for
future molecular studies, according Duszynski &
Gardner (1991). Both samples and the renal tissue
slides were deposited in the Parasitology Collection of
the Laborato´rio de Coccı´dios e Coccidioses, at
UFRRJ, located at the Municipality of Seropedica in
the State of Rio Janeiro, Brazil. Photovouchers and
line drawings are deposited and available (http://r1.
ufrrj.br/lcc) as well. Photographs of the host specimen
are deposited in the same collection. The repository
number is 53/2014.
Fig. 2 Photomicrographs of life-cycle stages of Klossiella tejerai in renal tissue from the Brazilian common opossum Didelphis aurita.
A–B, Macrogametes contained within parasitophorous vacuoles; C, Macrogamete and microgamete in syzygy within parasitophorous
vacuole; D–E, Early sporonts within parasitophorous vacuoles; F–G, Late budding sporonts within parasitophorous vacuoles; H, Oo¨cyst
with free mature sporoblasts/sporocysts; I, Macrogamete (right) and early sporont (middle) within parasitophorous vacuoles and oo¨cyst
with free mature sporoblasts/sporocysts (left). Scale-bars: 10 lm
Syst Parasitol (2014) 89:83–89 85
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Site of infection: Epithelium of the renal tubules.
Description (Figs. 1, 2)
Exogenous stages
Sporocysts ellipsoidal, 19–22 9 12–14 (20.4 9 12.7);
length/width (L/W) ratio 1.5–1.8 (1.6) (Fig. 1). Spo-
rocyst residuum present, composed of scattered
spherules. Sporozoites 12–14 (13), with anterior and
posterior refractile bodies and central nucleus.
Endogenous stages
Endogenous stages in parasitophorous vacuoles within
renal epithelial cells located near the corticomedullary
junction. Macrogametes subspheroidal to ovoidal,
7–11 9 5–9 (8.6 9 7.2), with basophilic nucleus and
contained within a subspheroidal to irregular parasitoph-
orous vacuole, 14–29 9 14–22 (22.6 9 18.3) (Fig. 2A–
B, I). Macrogamete in syzygy with microgametes in
some cases (Fig. 2C). Microgametes subspheroidal to
ovoidal, 5–6 9 3–5 (5.3 9 3.8), with basophilic
Table 1 Comparative morphology of Klossiella tejerai recovered from New World opossums
Host Didelphis aurita (Wied-
Neuwied)
Didelphis marsupialis
(L.)
Marmosa demerarae
(Thomas)
D. marsupialis (L.)
Reference Present study Scorza et al. (1957) Boulard (1975) Edgcomb et al.
(1976)
Macrogamete
Shape Subspherical to ovoidal – Subspherical to ovoidal –
Size 7–11 9 5–9 (8.6 9 7.2) (8 9 6) (12) 4–14 (9)
Parasitophorous vacuole
size
14–29 9 14–22
(22.6 9 18.3)
– – –
Microgamete
Shape Subspheroidal to ovoidal – Ovoidal –
Size 5–6 9 3–5 (5.3 9 3.8) (6 9 2) (9 9 6) –
Sporont
Shape Subspheroidal to irregular – – –
Size 15–31 9 14–21
(21.3 9 16.4)
up to 27 – 25–39 (28)
Number of nuclei 8–13 (10) – – –
Sporoblast/Sporocyst
Shape Ellipsoidal – – –
Size 10–13 9 6–9 (11.4 9 6.8) (12 9 9) (13.7 9 9) 14–17 (16)
Number per oo¨cyst 12–30 (18) (18) 16–22 –
Oo¨cyst
Shape Irregular – – –
Size 57–103 9 36–57
(71.6 9 47.2)
– (80 9 40) –
Urinary sporocysts/
sporozoites
Shape Ellipsoidal – – –
Size 19–22 9 12–14
(20.4 9 12.7)
– – –
Length/width ratio 1.5–1.8 (1.6) – – –
Sporocyst residuum Granular and diffuse – Present –
Number of sporozoites 12–14 (13) (12) 14–22 –
Refractile body 2, refringent, in both ends – – –
Nucleus Refringent and central – – –
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nucleus. Sporonts subspheroidal to irregular, 15–31 9
14–21 (21.3 9 16.4), dotted circumferentially with 8–13
(10) basophilic nuclei (Fig. 2D–E, I). Immature sporo-
blasts ellipsoidal, 5–6 9 3–4 (5.4 9 3.5) (Fig. 2F–G).
Mature sporoblasts/sporocysts 12–30 (18), ellipsoidal,
10–13 9 6–9 (11.4 9 6.8), with multiple dotted baso-
philic nuclei (Fig. 2H). Oo¨cysts irregular, 57–103 9
36–57 (71.6 9 47.2) (Fig. 2H–I).
Discussion
The description of K. tejerai of the current work confers
with the original description of Scorza et al. (1957) in
all characteristic features which were compared. The
descriptions of Edgcomb et al. (1976) and Boulard
(1975) had some divergences such as the measures of
the microgamete and sporoblast/sporocyst and the
number of sporozoites per sporocyst (Table 1).
Additionally, Edgcomb et al. (1976) and Boulard
(1975) observed merogonic stages (schizonts). How-
ever, in the current work and in the original description
of Scorza et al. (1957) merogonic stages in the renal
histology were not observed. The photomicrographs
of Edgcomb et al. (1976) are confusing and may have
been misinterpreted. In contrast, the meronts and
merozoites observed by Boulard (1975) are evident.
Scorza et al. (1957) suggested that the merogonic
stage should occur in other organs of the host, such as
the lungs, spleen, pancreas, testicles, etc. Thus, further
studies are needed to detail the merogonic stage of K.
tejerai and/or confirm that the specimens observed by
Boulard (1975) and Edgcomb et al. (1976) are K.
tejerai or another species.
In general, coccidiosis is an important disease that
affects the health, physiology and behavior of the
hosts. The immunity against coccidia develops
depending on the number of oo¨cysts/sporocysts
ingested; however, generally this immunity does not
prevent re-infection. In adult animals balance is
maintained between the constant re-infection and the
degree of immunity (Hunsaker, 1977; Aguilar et al.,
2008). In this sense, coccidiosis in wildlife in a habitat
without environmental impacts is rarely a significant
Fig. 3 Geographic ranges of some New World opossums, according IUCN (2014). A, Hosts for Klossiella tejerai are the didelphid
opossums Didelphis marsupialis, Marmosa demerarae and Didelphis aurita. Didelphis marsupialis is not sympatric with D. aurita;
however, M. demerarae is sympatric with D. marsupialis and D. aurita; B, Didelphis albiventris is another didelphid opossum with
wide geographic range sympatric with D. marsupialis and D. aurita which could disperse K. tejerai for Didelphis spp. and other New
World opossums
Syst Parasitol (2014) 89:83–89 87
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problem; on the other hand, epizootics should occur
when environmental disturbances and/or anthropo-
genic factors contribute to change the behavior and/or
mainly stressing the wild animals, leading to immun-
odepression. In this context, coccidia and coccidiosis
in wildlife, such as K. tejerai in D. aurita, assume role
of biomarkers of anthropization and/or environmental
disturbance (Giraudeau et al., 2014). In the current
work, the low prevalence, besides of the positive
opossum to be apparently healthy, demonstrate that the
location of capture, although anthropized, is favorable
to the demands of its ecological niche. On another point
of view, the generalist habit of the opossums may have
allowed its adaptation in an anthropized environment.
In another aspect, the coccidia are biomarkers of
transmission and, therefore, dispersion (Berto et al.,
2014b). Klossiella tejerai have been reported from only
two species of New World opossums. The route of
infection of this coccidian species is urine-oral; therefore,
the coccidial transmission should be usual among
sympatric opossums whichhave close ecological niches
favouring rapid transmission, since the sporocysts are not
resistant to desiccation, solar radiation, and other envi-
ronmental factors, due to thin sporocyst wall. Figure 3
shows the geographic ranges of these opossums to assist
in understanding the dynamics of dispersal of K. tejerai in
the New World. The direct transmission of K. tejerai
from D. marsupialis to D. aurita is unlikely, because they
are not sympatric. In contrast, M. demerarae is sympatric
with both D. marsupialis and D. aurita (Fig. 3A);
therefore, M. demerarae may potentially transmit K.
tejerai for these two hosts, and other sympatric suscep-
tible hosts. In this sense, Didelphis albiventris (Lund)
may be a potential susceptible host, which has wide
geographic range in South America and could disperse K.
tejerai for Didelphis spp. and other New World opossums
(Fig. 3B).
Finally, the current work is based on the concept of
intra-family specificity proposed by Duszynski &
Wilber (1997) which allows new hosts of the same
family. Therefore, considering Didelphidae as host-
family for K. tejerai with only two host species, D.
aurita becomes the third host species.
Acknowledgements This study was supported by grants from
the Fundac¸a˜o Carlos Chagas Filho de Amparo a` Pesquisa do
Estado do Rio de Janeiro (FAPERJ) to B. P. Berto (E-26/
110.987/2013).
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	Coccidial dispersion across New World marsupials: Klossiella tejerai Scorza, Torrealba & Dagert, 1957 (Apicomplexa: Adeleorina) from the Brazilian common opossum Didelphis aurita (Wied-Neuwied) (Mammalia: Didelphimorphia)
	Abstract
	Introduction
	Materials and methods
	Results
	Description (Figs. 1, 2)
	Discussion
	Acknowledgements
	References