Original Article

ISSN 1984-2961 (Electronic)

www.cbpv.org.br/rbpv
Braz. J. Vet. Parasitol., Jaboticabal, v. 28, n. 3, p. 395-402, july.-sept. 2019
Doi: https://doi.org/10.1590/S1984-29612019051

Toxoplasma gondii in cetaceans of Brazil: a histopathological

and immunohistochemical survey
Toxoplasma gondii em cetáceos do Brasil: estudo histopatológico e imuno-histoquímico
Samira Costa-Silva1* ; Carlos Sacristán1; Omar Gonzales-Viera2; Josué Díaz-Delgado1;
Angélica María Sánchez-Sarmiento1; Juliana Marigo1; Kátia Regina Groch1; Vitor Luz Carvalho3;
Ana Carolina Ewbank1; Adriana Castaldo Colosio4; Milton César Calzavara Marcondes4;
Ana Carolina Oliveira de Meirelles3; Carolina P. Bertozzi5; José Lailson-Brito6; Alexandre de Freitas Azevedo6;
Valéria Ruoppolo1; Larissa Oliveira7,8; Paulo Henrique Ott7,9; José Luiz Catão-Dias1
1
Laboratório de Patologia Comparada de Animais Selvagens, Departamento de Patologia, Faculdade de Medicina Veterinária e
Zootecnia, Universidade de São Paulo – USP, São Paulo, SP, Brasil
2
Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA, USA
3
Associação de Pesquisa e Preservação de Ecossistemas Aquáticos – AQUASIS, Caucaia, CE, Brasil
4
Instituto Baleia Jubarte, Caravelas, BA, Brasil
5
Biopesca, Instituto de Biociências, Universidade Estadual Paulista – UNESP, Campus do Litoral Paulista, São Vicente, SP, Brasil
6
Laboratório de Mamíferos Aquáticos e de Bioindicadores “Profa Izabel M. G. do N. Gurgel” – MAQUA, Faculdade de
Oceanografia, Universidade do Estado do Rio de Janeiro – UERJ, Rio de Janeiro, RJ, Brasil
7
Grupo de Estudos de Mamíferos Aquáticos do Rio Grande do Sul – GEMARS, Torres, RS, Brasil
8
Laboratório de Ecologia de Mamíferos, Universidade do Vale do Rio dos Sinos – UNISINOS, São Leopoldo, RS, Brasil
9
Laboratório de Biodiversidade e Conservação, Universidade Estadual do Rio Grande do Sul – UERGS, Unidade Litoral Norte,
Osório, RS, Brasil

Received February 14, 2019

Accepted May 24, 2019

Abstract
Toxoplasmosis is a parasitic disease caused by the protozoan Toxoplasma gondii. In cetaceans, T. gondii infection is a
significant cause of morbidity and mortality. Despite the worldwide range and broad cetacean host record of T. gondii
infection, there is limited information on toxoplasmosis in cetaceans from the Southern hemisphere. We investigated
the occurrence of T. gondii by histopathology and immunohistochemistry in tissue samples of 185 animals comprising
20 different cetacean species from Brazil. Three out of 185 (1.6%) animals presented T. gondii-associated lesions: a captive
killer whale Orcinus orca, a free-ranging common bottlenose dolphin Tursiops truncatus and a free-ranging Guiana dolphin
Sotalia guianensis. The main lesions observed in these animals were necrotizing hepatitis, adrenalitis and lymphadenitis
associated with protozoal cysts or extracellular tachyzoites presenting immunolabeling with anti-T. gondii antibodies.
This study widens the spectrum of species and the geographic range of this agent in Brazil, and provides the first reports
of T. gondii infection in a captive killer whale and in a free-ranging common bottlenose dolphin in South America.
Keywords: Toxoplasmosis, mortality, stranding, South America, protozoan infection, marine mammal.

Resumo
Toxoplasmose é uma doença parasitária causada pelo protozoário Toxoplasma gondii. A infecção por T. gondii é uma
causa significativa de morbidade e mortalidade, nos cetáceos. Apesar da abrangência mundial e amplo registro de espécies
de cetáceos infectadas por T. gondii, informações sobre toxoplasmose em cetáceos do hemisfério sul são limitadas. Neste
estudo pesquisou-se por meio de histopatologia e imuno-histoquímica a ocorrência de T. gondii em amostras de tecido
de 185 animais, compreendendo 20 diferentes espécies de cetáceos que ocorrem no Brasil. Três dos 185 (1,6%) animais
apresentaram lesões associadas a T. gondii: uma orca Orcinus orca mantida em cativeiro, um golfinho-nariz-de-garrafa

*Corresponding author: Samira Costa-Silva. Laboratório de Patologia

Comparada de Animais Selvagens, Departamento de Patologia, Faculdade
de Medicina Veterinária e Zootecnia, Universidade de São Paulo – USP,
Av. Orlando Marques de Paiva, 87, Butantã, CEP 05508-270, São Paulo, SP,
Brasil. e-mail: [email protected]

This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution,
and reproduction in any medium, provided the original work is properly cited.
396 396/402 Costa-Silva, S. et al. Braz. J. Vet. Parasitol.

Tursiops truncatus e um boto-cinza Sotalia guianensis de vida livre. As principais lesões observadas nesses animais foram
hepatite, adrenalite e linfadenite necrotizantes associadas a cistos protozoários ou taquizoítos extracelulares, marcados
com anticorpos anti-T. gondii. O presente estudo amplia o espectro de espécies susceptíveis a esse agente e o seu alcance
geográfico no Brasil, fornecendo o primeiro relato da infecção por T. gondii em uma orca mantida em cativeiro e em
um golfinho-nariz-de-garrafa de vida livre na América do Sul.
Palavras-chave: Toxoplasmose, mortalidade, encalhe, América do Sul, infecção por protozoários, mamífero
marinho.

Introduction 2013). We hypothesized that T. gondii infection could be

present in a wider number of cetacean species in Brazil than
Toxoplasma gondii is a zoonotic intracellular coccidian protozoan previous studies have suggested. Thus, the aim of this study
of the phylum Apicomplexa (DUBEY, 2008). This protozoan was to evaluate retrospectively the occurrence of T. gondii
is one of the most common parasites of warm-blooded animals infection in captive and free-ranging cetaceans employing
(DUBEY et al., 2004), and was described for the first time in histopathological and immunohistochemical (IHC) analyses
Brazil and in Tunisia, in 1908 (NICOLLE & MANCEAUX, on tissue samples obtained from a large marine mammal tissue
2009; SPLENDORE, 2009). The first record of toxoplasmosis bank in Brazil.
in cetaceans was in a Guiana dolphin Sotalia guianensis from Rio
de Janeiro state, Brazil, in the 1970’s (BANDOLI & OLIVEIRA,
1977). Since then, the exposure and infection by this protozoan
Materials and Methods
has been detected through serological, pathological and molecular
We evaluated formalin-fixed paraffin-embedded tissue samples
techniques in a wide variety of captive and free-ranging cetaceans
stored at the Marine Mammal Tissue Bank of the Laboratory of
worldwide (INSKEEP et al., 1990; OMATA et al., 2006;
Wildlife Comparative Pathology, School of Veterinary Medicine
MAZZARIOL et al., 2012; GONZALES-VIERA et al., 2013;
and Animal Science, University of São Paulo, Brazil. The samples
IQBAL et al., 2018). Most of these studies have covered individual
included in the study came from partial or complete standard
cases or small groups of cetaceans (DI GUARDO et al., 2010;
necropsies conducted on odontocetes and mysticetes between
GONZALES-VIERA et al., 2013). Toxoplasmosis is a significant
1988 and 2014, provided by partner institutions attending marine
cause of morbidity and may lead to stranding and death, and is
mammal strandings, rehabilitation and incidental captures along
considered one of the most important emerging diseases in cetaceans
worldwide (VAN BRESSEM et al., 2009; DI GUARDO et al., the Brazilian coast. Tissues from 185 individuals of 20 different
2010; GONZALES-VIERA et al., 2013; BIGAL et al., 2018). cetacean species (Table 1) were evaluated. Among the evaluated
The epidemiology of T. gondii transmission has been established samples, three were from originally wild animals kept in captivity
in several terrestrial animals (e.g., cats, livestock), and to a lesser in two different amusement parks of the São Paulo state (Brazil):
extent, aquatic species (e.g., shellfish), including transplacental a killer whale Orcinus orca from Iceland kept in captivity in the
transmission in susceptible pregnant animals (TENTER et al., 1980’s, an Amazon river dolphin from the Amazon Basin, and
2000; CATÃO-DIAS et al., 2013; JARDINE & DUBEY, 2002; a common bottlenose dolphin Tursiops truncatus of unknown
MILLER et al., 2008). In terrestrial animals, infection also occurs origin. The remaining animals were all free-ranging species that
through ingestion of tissue cysts or contaminated food and/or stranded along the Brazilian coast, mostly from the southern
water containing infecting sporulated oocysts released in feline region (43.9%; 80/182), followed by the southeastern (30.2%;
feces. However, the transmission of T. gondii in cetaceans has 55/182), and northeastern (25.8%; 47/182) regions (Figure 1).
not been fully elucidated. One major source of transmission in The examined tissues were: liver, kidney, lung, spleen, skeletal
cetaceans could be the polluted marine environments in proximity muscle, lymph node, adrenal gland, brain, uterus, intestines,
with the coast, e.g., run-off of cat feces or soil contaminated heart and eye.
with T. gondii oocysts from rivers, polluted effluents and ballast Formalin-fixed paraffin-embedded tissue sections from the
water from ships (VAN BRESSEM et al., 2009). In utero aforementioned organs were sectioned at 5 µm thickness and
transmission of T. gondii has been described in Risso’s dolphin stained with H&E. Additionally, special histochemical techniques
Grampus griseus and Indo-Pacific bottlenose dolphin Tursiops - periodic acid-Schiff (PAS), Giemsa and Grocott’s methenamine
aduncus (JARDINE & DUBEY, 2002; RESENDES et al., silver stains - were used in selected tissue sections to improve
2002). For offshore marine ecosystems, T. gondii transmission visualization of protozoan structures compatible with T. gondii
patterns are still unclear. (LUNA, 1992; POPPER et al., 1960).
The occurrence of T. gondii in marine mammals remains a Immunohistochemical analysis for T. gondii was performed
poorly understood phenomenon, particularly in Brazil, where very on selected tissues for which T. gondii has demonstrated
limited information exists. Exposure to T. gondii was reported tropism in cetaceans (i.e., liver, kidney, lung, brain and lymph
in Amazon river dolphins Inia geoffrensis (SANTOS et al., nodes) (INSKEEP et al., 1990; MIKAELIAN et al., 2000;
2011), but to this date, only one cetacean species is known to RESENDES et al., 2002; VAN BRESSEM et al., 2009;
be infected by T. gondii in Brazilian waters: the Guiana dolphin GONZALES-VIERA et al., 2013). Briefly, 3 μm -thick sections
(BANDOLI & OLIVEIRA, 1977; GONZALES-VIERA et al., were deparaffinized and rehydrated through a series of graded
v. 28, n. 3, july.-sept. 2019 Toxoplasma gondii in cetaceans of Brazil 397/402 397

Table 1. Cetacean species evaluated in Brazil for Toxoplasma gondii by immunohistochemistry.

Number of Sexb
Species
individuals M F U
Franciscana dolphin (Pontoporia blainvillei) 102 57 41 4
Guiana dolphin (Sotalia guianensis)a 27 14 11 2
Dwarf sperm whale (Kogia sima) 7 4 3 0
Atlantic spotted dolphin (Stenella frontalis) 6 6 0 0
Spinner dolphin (Stenella longirostris) 5 4 1 0
Clymene dolphin (Stenella clymene) 6 3 3 0
Common bottlenose dolphin (Tursiops truncatus)a 4 2 1 1
Melon-headed whale (Peponocephala electra) 5 3 2 0
Rough-toothed dolphin (Steno bredanensis) 3 2 1 0
Short-finned pilot whale (Globicephala macrorhynchus) 3 3 0 0
Sperm whale (Physeter macrocephalus) 3 1 2 0
Pygmy sperm whale (Kogia breviceps) 2 2 0 0
Humpback whale (Megaptera novaeangliae) 2 2 0 0
Fraser’s dolphin (Lagenodelphis hosei) 2 0 2 0
Striped dolphin (Stenella coeruleoalba) 2 0 2 0
Killer whale (Orcinus orca)a 2 1 1 0
Short-beaked common dolphin (Delphinus delphis) 1 1 0 0
Risso’s dolphin (Grampus griseus) 1 0 1 0
Gervais’ beaked whale (Mesoplodon europaeus) 1 0 1 0
Amazon river dolphin (Inia geoffrensis) 1 0 0 1
TOTAL 185 105 72 8
a
Species with positive results; bM = male; F = female; U = unknown.

alcohols. Toxoplasma gondii antigen was retrieved by heating tissue

sections in citrate buffer (pH=7.0) solution for seven minutes at
90 °C. The sections were blocked with 1% normal rabbit serum
in PBS for 30 minutes followed by overnight incubation with a
polyclonal goat anti-T. gondii primary antibody (1 in 400 dilution;
VMRD Inc; Pullman). This antibody does not cross-react with
Neospora caninum (CORBELLINI et al., 2002). The sections were
washed in PBS and incubated for 30 minutes with biotinylated
polyclonal anti‑goat secondary antibody (1 in 600 dilution;
Dako) as previously described (DI GUARDO et al., 2010).
Amplification of the immunologic reaction was based on an
avidin-biotin-peroxidase complex method (Elite ABC kit, Vector
laboratories), following manufacturer’s instructions. Labeling
was ‘visualized’ with 3-amino-9-ethyl-carbazole (Sigma) and/or
diaminobenzidine (DAB D-5637; Sigma), and sections were
counterstained with Mayer’s haematoxylin. Tissue sections in
which the primary antibodies were replaced by phosphate buffered
saline or nonimmune homologous serum served as negative
controls. Sections of the adrenal gland of a Guiana dolphin
infected by T. gondii (GONZALES-VIERA et al., 2013) were
used as positive control. Morbillivirus immunohistochemistry
was also performed on selected tissue sections of T. gondii positive
cases to rule out potential coinfection, following a previously
published protocol (SALIKI et al., 2002; GROCH et al.,
2014), using a commercial monoclonal antibody against the
nucleoprotein of Canine distemper virus (VMRD Inc.), known
Figure 1. Geographic distribution along the coastline of Brazil of to cross-react with Cetacean morbillivirus. Positive tissue samples
Toxoplasma gondii- positive (green triangle) and -negative (red circle) of an infected Guiana dolphin were selected as positive control
animals, according to immunohistochemistry. (GROCH et al., 2014).
398 398/402 Costa-Silva, S. et al. Braz. J. Vet. Parasitol.

Results T. gondii-associated lesions in these animals follows. Additional

information regarding the microscopic findings of positive animals
Three of 185 (1.6%) animals were IHC-positive for T. gondii-: is available in Table 2. All three T. gondii positive animals were
a captive killer whale, a free-ranging common bottlenose dolphin, IHC-negative for morbillivirus. No PCR techniques were employed
and a free-ranging Guiana dolphin. These animals presented in these cases because frozen tissue samples were not available.
a variety of toxoplasmosis-compatible lesions, with positively Case No 1 (MM#452) was a 5-6 years-old juvenile (2,500 kg)
labeled protozoal cysts and free tachyzoites. The occurrence of male killer whale captured in Iceland in 1983, and one year later
T. gondii in free-ranging species was 1.1% (2/182), whereas the brought into an oceanarium located in the city of São Paulo, São
occurrence in captive animals was 33.3% (1/3). A summary of Paulo state, Brazil. The animal remained in this facility until its

Table 2. Microscopic lesions in Toxoplasma gondii-positive animals in Brazil.

Microscopic lesions
Organ
Case 1 (killer whale) Case 2 (common bottlenose dolphin) Case 3 (Guiana dolphin)
Liver Moderate randomly multifocal to Moderate to marked randomly NA
coalescent necrotizing hepatitis with multifocal to coalescent necrotizing
protozoan cysts, compatible with hepatitis with protozoan cysts,
T. gondii. T. gondii-positive by IHC. compatible with T. gondii, and
predominantly mononuclear
infiltrate, mild hepatic congestion and
hemorrhage. T. gondii-positive by IHC.
Kidney Acute multifocal interstitial nephritis Mild multifocal chronic tubular ectasia NA
with mild to moderate mixed infiltrate, and glomerular cyst formation.
mild to moderate membranous
glomerulonephritis.
Adrenal gland NA NA Marked multifocal acute necrotizing
adrenalitis associated with numerous
round to oval protozoal cysts and
extracellular tachyzoites compatible with
T. gondii. T. gondii-positive by IHC.
Lung Moderate to marked multifocal acute Moderate to marked multifocal acute NA
fibrinosuppurative and hemorrhagic fibrinosuppurative bronchopneumonia
bronchopneumonia with protozoan cysts with hemorrhage.
(compatible with T. gondii) and bacteria.
T. gondii-positive by IHC.
Stomach Mild to moderate multifocal to NA NA
coalescent granulocytic gastritis.
Intestine Mild to moderate multifocal Autolysis NA
granulocytic lymphadenitis, mild to
moderate diffuse mixed enteritis with
crypt microabscesses (cryptitis).
Muscle Mild multifocal acute segmental myocyte NA NA
degeneration and necrosis.
Thyroid Mild multifocal to coalescent NA NA
lymphoplasmacytic thyroiditis.
Lymph node Marked necrosuppurative lymphadenitis NA NA
with protozoal cysts (compatible with
T. gondii), and lymphoid depletion with
lymphocytolysis. T. gondii-positive by
IHC.
Vascular Acute fibrinoid vasculitis with NA NA
mononuclear infiltrate.
Eye NA NA No significant findings observed.
NA = not available.
v. 28, n. 3, july.-sept. 2019 Toxoplasma gondii in cetaceans of Brazil 399/402 399

death, on March 7th, 1988. No clinical data were available. Samples and lymphoid necrosis, interstitial pneumonia, adrenal necrosis,
of lungs, liver, spleen, kidneys, skeletal muscle, glandular stomach, and non-suppurative encephalitis and meningoencephalitis
large intestine, thyroid glands, and lymph nodes were collected with bradyzoites and free tachyzoites (INSKEEP et al., 1990;
upon necropsy. The only reported macroscopic finding was a MIGAKI et al., 1990; MIKAELIAN et al., 2000; RESENDES et al.,
renal cyst. Microscopically, the main lesions included moderate 2002; VAN BRESSEM et al., 2009; GONZALES-VIERA et al.,
to marked, multifocal, acute fibrinosuppurative and hemorrhagic 2013; ROE et al., 2013). In T. gondii‑positive cases, all tested
bronchopneumonia with protozoan cysts, moderate randomly tissues were negative for morbillivirus, suggesting that T. gondii
multifocal to coalescent necrotizing hepatitis with protozoan cysts possibly acted as a primary agent, as previously observed
(compatible with T. gondii), marked necrosuppurative lymphadenitis, (DI GUARDO et al., 2010; GONZALES-VIERA et al., 2013;
acute multifocal interstitial nephritis with mild‑moderate mixed ROE et al., 2013). The scarce number of brain samples available
infiltrate, mild to moderate, multifocal, chronic membranous for this study (n = 4) probably prevented the identification of
glomerulonephritis, and acute fibrinoid vasculitis. T. gondii-associated lesions in nervous tissue, which is commonly
Case No 2 (MM#178) was a juvenile male common bottlenose involved in cetacean toxoplasmosis cases (DI GUARDO et al.,
dolphin of 2.43 m total body length, that stranded alive and died 2010; SIERRA et al., 2014).
shortly after stranding, on October 29th, 2001, in Lagos Region, In this study, we evaluated cetacean samples from geographic
Rio de Janeiro state, Brazil (22°56’S, 42°19’W). Samples of lungs, areas that correspond to a large portion of the Brazilian coastline,
liver, kidneys, and large intestine were collected upon necropsy. most of them coastal species, mainly franciscanas (Pontoporia
No gross findings were reported. The main microscopic findings blainvillei) (55.0%) and Guiana dolphins (14.5%). Coastal
observed were moderate to severe, multifocal random to coalescing cetaceans, considered suitable sentinels of the marine environmental
necrotizing hepatitis with protozoan cysts compatible with T. gondii ecosystem and important indicators of marine pollution, are
and moderate to marked, multifocal, acute fibrinosuppurative usually more exposed to anthropogenic activities (WELLS et al.,
bronchopneumonia. 2004; BOSSART, 2011; MOURA et al., 2014). From these,
Case No 3 (MM#67) was a 1.86 m-long adult, female Guiana we found histological and immunohistochemical evidences of
dolphin that stranded dead on March 3th, 1998, in Paranaguá Bay, toxoplasmosis in one Guiana dolphin. This is the third report of
Paraná state, Brazil (25°31’S, 48°30’W). No necropsy records toxoplasmosis in this species (BANDOLI & OLIVEIRA, 1977;
were available. Samples of eye and adrenal glands were evaluated. GONZALES‑VIERA et al., 2013). This animal presented severe
Microscopically, the main finding was marked, multifocal, adrenalitis, similar to what was reported by Gonzales-Viera et al.
acute necrotizing adrenalitis with numerous protozoal cysts and (2013) in another Guiana dolphin from the same area (Paranaguá
extracellular tachyzoites compatible with T. gondii, and further Bay) that died in the same year (1998). Given the marked pattern
highlighted with PAS and Giemsa stains. of site fidelity presented by Guiana dolphins (MOURA et al.,
2014), this result suggests a common source of exposure to
T. gondii and a possible negative impact of toxoplasmosis on this
Discussion population. Although the low occurrence of T. gondii infection
found in the present study does not allow us to infer further
Despite the widespread geographical range of T. gondii infection, on this protozoan’s geographic distribution, the location of the
in Brazil, the current knowledge regarding cetacean toxoplasmosis Guiana dolphin case and previous reports suggest that infection
is limited to few species and studies (BANDOLI & OLIVEIRA, may occur at higher rates in coastal areas of Brazil. Coastal areas
1977; SANTOS et al., 2011; GONZALES-VIERA et al., 2013). usually receive freshwater run-offs and terrestrial biologic pollutants,
Based on microscopical and immunohistochemical examinations, contributing to the presence of T. gondii (SHAPIRO et al., 2015).
we provide the first description of T. gondii in a common These results also suggest areas in which future research efforts
bottlenose dolphin of South America, and in a captive killer whale. should be concentrated.
Furthermore, the identification of toxoplasmosis in a Guiana Concerning killer whales, positive serology for T. gondii
dolphin corroborates with previous observations regarding this was reported in a killer whale calf that stranded alive in Japan,
species and geographic area, the Paranaguá Bay. On the other hand, in 1988 (MURATA et al., 2004), and T. gondii infection was
in spite of the high prevalence of T. gondii antibodies reported in identified by molecular techniques in a stranded killer whale from
free-ranging Amazon river dolphins (SANTOS et al., 2011), our the Northeastern Pacific, in California (GIBSON et al., 2011).
sampled captive Amazon river dolphin was T. gondii- negative. However, these reports lacked detailed pathological examinations.
We detected protozoal cysts and free tachyzoites of T. gondii In the present study, we provide the first histopathological and
by IHC and observed histological changes within multiorgan immunohistochemical evidences of toxoplasmosis in this species.
necroinflammatory foci, e.g., necrotizing hepatitis, necrosuppurative Based on the severity and extent of characteristic multiorganic
lymphadenitis (killer whale), necrotizing hepatitis (common bottlenose necroinflammatory foci, we believe that T. gondii played a major
dolphin), and necrotizing adrenalitis (Guiana dolphin). These lesions role in the death of the evaluated killer whale. In this case, potential
are in agreement with previous reports of T. gondii infection in environmental sources of T. gondii infectious forms may include
odontocetes (CRUICKSHANK et al., 1990; RESENDES et al., water and/or food contamination in one or more of the following
2002; DI GUARDO & MAZZARIOL, 2013) and mysticetes scenarios: (1) the facility’s water system (e.g., used to clean fish
(MAZZARIOL et al., 2012). The main lesions observed in cetacean and the enclosure, and fill the tank), especially considering that
cases of toxoplasmosis are necrotizing hepatitis, lymphadenitis sporulated oocysts of T. gondii are viable in saltwater for at least
400 400/402 Costa-Silva, S. et al. Braz. J. Vet. Parasitol.

two years (AUBERT & VILLENA, 2009); (2) contaminated food

(e.g., fish); and (3) contaminated enclosure (e.g., contaminated
fomites, personnel and/or presence of roaming cats). These findings
highlight the importance of proper hygiene and husbandry in
captive cetaceans, including properly treated water and reliable
food sources free of T. gondii sporulated oocysts. Although
unlikely, considering the physiopathology of toxoplasmosis in
highly susceptible or ‘less adapted’ hosts (CATÃO-DIAS et al.,
2013) - which could be the case for the killer whale - one should
also consider the possibility of infection prior to the specimen’s
capture in the wild (e.g., transplacental or transmammary) or
initial period in captivity (1984-1988). In that case, the infection
would have been quiescent until being activated by the stress of
captivity and/or other predisposing factors (e.g., concomitant
diseases) (LINDSAY et al., 2003; AUBERT & VILLENA, 2009;
MAZZARIOL et al., 2012).
Toxoplasma gondii infections in common bottlenose dolphins
have been observed worldwide; in the Mediterranean Sea and the
Atlantic Ocean (CRUICKSHANK et al., 1990; INSKEEP et al.,
1990; DI GUARDO et al., 1995; DUBEY et al., 2008; PRETTI et al.,
2010; PROFETA et al., 2015; BIGAL et al., 2018), and also in
captive specimens (DUBEY et al., 2009). Furthermore, it has also
been detected in a related species from the Pacific Ocean - the
Indo-Pacific bottlenose dolphin (JARDINE & DUBEY, 2002).
However, this is the first report of T. gondii in an Atlantic bottlenose
dolphin from South America (Figure 2), widening the geographic
range of T. gondii occurrence for this species.
Because cetacean species’ susceptibility to T. gondii infection is
unknown, non-exposure and natural resistance to this agent should
be considered in negative cases (DI GUARDO & MAZZARIOL,
2013). Omata et al. (2006) proposed that killer whales from Japan
might be resistant to T. gondii; however, the evidence provided
seemed slight because none of these animals presented antibodies
against the agent.
Finally, this is the first report of a large-scale T. gondii survey
based on histopathological and immunohistochemical examinations
in cetaceans of Brazil. This study widens the spectrum of
T. gondii‑susceptible species and geographic range of this agent
in Brazil and presents the first report of T. gondii-infection in a
captive killer whale, and in a free-ranging Atlantic bottlenose
dolphin from South America. Furthermore, it corroborates with
previous observations of toxoplasmosis in Guiana dolphin and the
geographic area of Paranaguá Bay. Further studies are warranted
to characterize the circulating strains and clarify the distribution
of this protozoan, its transmission route(s), unsolved pathogenetic
mechanisms, cetacean host-specific susceptibilities and potential
implications to the conservation of cetacean species of Brazil and
Figure 2. Microscopic lesions and immunohistochemical findings in mechanisms involved in human exposure.
Toxoplasma gondii positive animals. (A) Atlantic bottlenose dolphin (Tursiops
truncatus). Liver, necrotizing hepatitis (asterisk), H&E, scale bar = 200 μm;
(B) Killer whale (Orcinus orca). Lung, bronchopneumonia (arrows) with Acknowledgements
alveolar infiltrate (asterisk), H&E, scale bar = 200 μm; (C.1) Guiana
dolphin (Sotalia guianensis). Adrenal gland, there is focally extensive lytic We thank Leonardo L. Wedekin, Jorge Oyakawa and Sândara
necrosis with multiple protozoal cysts (arrows). IHC for T. gondii, Mayer’s Sguario for their technical support. We also thank the Aquário
hematoxylin counterstaining, scale bar = 500 μm; (C.2) Detailed view of de Santos, Associação de Pesquisa e Preservação de Ecossistemas
T. gondii protozoal cyst containing numerous bradyzoites. IHC for T. gondii, Aquáticos (AQUASIS), Instituto Baleia Jubarte, Instituto de
Mayer’s hematoxylin counterstaining, scale bar = 25 μm. Pesquisas de Cananéia (IPeC), Grupo de Estudos de Mamíferos
v. 28, n. 3, july.-sept. 2019 Toxoplasma gondii in cetaceans of Brazil 401/402 401

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Mamíferos Aquáticos e Bioindicadores ‘Profa. Izabel Gurgel’ - WE, et al. Isolation of Toxoplasma gondii from bottlenose dolphins
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org/10.1645/GE-1444.1. PMid:18576793.
Janeiro (MAQUA), Museu Oceanográfico da FURG, Playcenter,
Projeto Biopesca - Universidade Estadual Paulista-UNESP, and Dubey JP, Lipscomb TP, Mense M. Toxoplasmosis in an elephant seal
Projeto Baleia Franca/Instituto Australis for providing the samples (Mirounga angustirostris). J Parasitol 2004; 90(2): 410-411. http://dx.doi.
and necropsy records. This study was co-financed by CAPES and org/10.1645/GE-155R. PMid:15165069.
FAPESP. The financial sources had no influence over the project’s Dubey JP, Mergl J, Gehring E, Sundar N, Velmurugan GV, Kwok
development. All procedures were performed in accordance with OC, et al. Toxoplasmosis in captive dolphins (Tursiops truncatus) and
the Ethical Committee of the School of Veterinary Medicine walrus (Odobenus rosmarus). J Parasitol 2009; 95(1): 82-85. http://dx.doi.
and Animal Sciences, University of São Paulo (process number: org/10.1645/GE-1764.1. PMid:19245284.
2551070144). JDD is the recipient of a post-doctoral fellowship by Dubey JP. The history of Toxoplasma gondii - The first 100 years. J Eukaryot
FAPESP (grant #2017/02223-8), and CNPq provided scholarship Microbiol 2008; 55(6): 467-475. http://dx.doi.org/10.1111/j.1550-
to J.L.C.-D (grant #305349/2015-5). 7408.2008.00345.x. PMid:19120791.
Gibson AK, Raverty S, Lambourn DM, Huggins J, Magargal SL, Grigg
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