93
Zoonotic infections in soil of recreational areas
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
Neotropical Helminthology
Neotropical Helminthology, 2024, vol. 18 (1) 93-102
ORIGINAL ARTICLE / ARTÍCULO ORIGINAL
FIRST RECORD OF PARASITISM BY
HEPATOZOON
MILLER, 1908
(APICOMPLEXA: HEPATOZOIDAE) IN ANURANS FROM THE
CAATINGA, BRAZIL
PRIMER REGISTRO DE PARASITISMO POR
HEPATOZOON
MILLER, 1908
(APICOMPLEXA: HEPATOZOIDAE) EN ANUROS DE LA
CAATINGA, BRASIL
PRI
MEIRO REGISTRO DE PARASITISMO POR
HEPATOZOON
MILLER, 1908
(APIC
OMPLEXA: HEPATOZOIDAE) EM ANUROS DE
CAATINGA, BRASIL
Carla Adrielle Costa de Carvalho
1
, Gabriela Felix-Nascimento
2
, Fabiano Matos Vieira
3,*
, Lucio
André Viana
4
& Leonardo Barros Ribeiro
1,5
ISSN Versión Impresa 2218-6425 ISSN Versión Electrónica 1995-1403
DOI: https://dx.doi.org/10.62429/rnh20241811782
Este artículo es publicado por la revista Neotropical Helminthology de la Facultad de Ciencias Naturales y Matemática, Universidad Nacional Federico
Villarreal, Lima, Perú auspiciado por la Asociación Peruana de Helmintología e Invertebrados Af nes (APHIA). Este es un artículo de acceso abierto,
distribuido bajo los términos de la licencia Creative Commons Atribución 4.0 Internacional (CC BY 4.0) [https:// creativecommons.org/licenses/by/4.0/
deed.es] que permite el uso, distribución y reproducción en cualquier medio, siempre que la obra original sea debidamente citada de su fuente original.
1
Programa de Pós-graduação em Ciências Veterinárias no Semiárido, Universidade Federal do Vale do São Francisco. Rodovia
BR-407, KM 12, Lote 543, S/N, Projeto de Irrigação Senador Nilo Coelho, Petrolina, Pernambuco 56300-000, Brasil.
² Laboratório de Anatomia dos Animais Domésticos e Silvestres (LAADS), Universidade Federal do Vale do São Francisco.
Rodovia BR-407, KM 12, Lote 543, S/N, Projeto de Irrigação Senador Nilo Coelho, Petrolina, Pernambuco 56300-000,
Brasil.
³ Laboratório de Microscopia e Lupas, Universidade Federal do Vale do São Francisco. Rodovia BR-407, KM 12, Lote 543,
S/N, Projeto de Irrigação Senador Nilo Coelho, Petrolina, Pernambuco 56300-000, Brasil.
4
Laboratório de Estudos Morfof siológicos e Parasitários, Departamento de Ciências Biológicas e da Saúde, Universidade
Federal do Amapá. Rodovia Juscelino Kubitscheck, KM 02, S/N, Jardim Marco Zero, Macapá, Amapá 68903-419, Brazil.
5
Colegiado de Ciências Biológicas, Universidade Federal do Vale do São Francisco. Rodovia BR-407, KM 12, Lote 543, S/N,
Projeto de Irrigação Senador Nilo Coelho, Petrolina, Pernambuco 56300-000, Brasil.
*
Corresponding author: fmatosvieira@gmail.com
Carla Adrielle C osta de Carvalho:
https://orcid.org/0000-0001-6286-8284
Gabriela Felix-Nascimento:
https://orcid.org/0000-0001-8623-9546
Fabiano Matos Vieira:
https://orcid.org/0000-0002-5220-7252
Lucio André Viana:
https://orcid.org/0000-0002-0932-0479
Leonardo Barros Ribeiro:
https://orcid.org/0000-0003-4491-0236
94
Carvalho
et al.
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
ABSTRACT
Tis study aimed to register for the frst time parasitism by
Hepatozoon
Miller, 1908 in two species of bufonid anurans
in areas of the Caatinga in Brazil, and to present morphometric analysis on the parasites and parasitized erythrocytes.
During helminthological studies on anurans from the Brazilian Caatinga in 2019, blood smears of three specimens of
Rhinella diptycha
(Cope, 1862) and three of
Rhinella granulosa
(Spix, 1824) in the municipality of Petrolina, state of
Pernambuco, Brazil were analyzed. To make blood smears, blood samples were collected intracardiacally or through
puncturing the ventral abdominal vein. Te smears were analyzed under a light microscope at 1000x magnifcation.
Morphometric dimensions of host cells and parasites were measured. Te only parasite forms encountered in the blood
of the two anuran species were intraerythrocytic gametocytes of
Hepatozoon
spp., only parasitizing erythrocytes, which
had a hypertrophied appearance. Two distinct morphotypes of
Hepatozoon
were found: one in each host species and
both of them diferent from morphotypes previously described and registered in Brazil. Terefore, the present study
constitutes the frst report of
Hepatozoon
sp. in anurans in the Caatinga biome and also forms the frst register of parasites
of this genus in
R. granulosa
in this country.
Keywords:
Amphibians – Bufonidae – Haemogregarines – Hemoparasites.
RESUMO
O estudo teve o objetivo registrar pela primeira vez o parasitismo por
Hepatozoon
Miller, 1908 em duas espécies de
anuros bufonídeos em áreas de Caatinga no Brasil, assim como a morfometria dos parasitos e eritrócitos parasitados.
Durante estudos helmintológicos em anuros da Caatinga brasileira em 2019 foram analisados esfregaços sanguíneos de
3 espécimes de
Rhinella diptycha
(Cope, 1862) e três
Rhinella granulosa
(Spix, 1824) no município de Petrolina, estado
de Pernambuco, Brasil. Para montagem desses esfregaços sanguíneos, foram utilizadas amostras de sangue coletado via
intracardíaca ou por punção da veia abdominal ventral.
A análise dos esfregaços foi realizada sob microscópio de luz
com a magnifcância de 1000x. Células hospedeiras e parasitos tiveram mensuradas dimensões morfométricas.
A
s únicas
formas parasitárias encontradas no sangue das duas espécies de anuros foram gametócitos intraeritrocíticos de
Hepatozoon
spp., parasitando somente eritrócitos que se apresentaram hipertrofados. Foram encontrados dois morfotipos distintos
de
Hepatozoon
,
um em cada espécie de hospedeiros, todos diferentes dos morfotipos já descritos e registrados no país.
Portanto, o presente estudo se constitui no primeiro relato de
Hepatozoon
sp. em anuros no bioma da Caatinga, além de
também ser o primeiro registro de parasitos desse gênero em
R. granulosa
no país.
Palavras-chave
: Anfbios – Bufonidae – hemogregarina – hemoparasitos
RESUMEN
El estudio tuvo como objetivo registrar por primera vez el parasitismo por
Hepatozoon
Miller, 1908 en dos especies de
anuros bufonídeos en áreas de Caatinga en Brasil, así como la morfometría de los parásitos y los eritrocitos parasitados.
Durante estudios helmintológicos en anuros de la Caatinga brasileña en 2019, se analizaron extendidos sanguíneos de
3 especímenes de
Rhinella diptycha
(Cope, 1862) y
Rhinella granulosa
(Spix, 1824) en el municipio de Petrolina, estado
de Pernambuco, Brasil. Para la preparación de los extentidos sanguíneos, se utilizaron muestras de sangre recolectadas
mediante punción intracardiaca o de la vena abdominal ventral. El análisis de los extentidos se realizó bajo un microscopio
de luz con una magnifcación de 1000x. Se midieron las dimensiones morfométricas de las células hospedadoras y los
parásitos. Las únicas formas parasitarias encontradas en la sangre de las dos especies de anuros fueron gametocitos
intraeritrocíticos de
Hepatozoon
spp., que parasitan solo a eritrocitos que muestran hipertrofa. Se encontraron dos
morfotipos distintos de
Hepatozoon
, uno en cada especie de hospedador, todos diferentes de los morfotipos ya descritos
y registrados en el país. Por lo tanto, el presente estudio constituye el primer informe de
Hepatozoon
sp. en anuros en
el bioma de la Caatinga, además de ser también el primer registro de parásitos de este género en
R. granulosa
en el país.
Palabras clave
: Anfbios – Bufonidae – hemogregarina – hemoparásitos
95
Hepatozoon
of anurans from Caatinga
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
INTRODUCTION
Te phylum Apicomplexa comprises lineages of obligate
parasites of great diversity, with over 6,000 described
species and possibly thousands yet to be discovered.
Among these, hemogregarines are the most frequent
hemoparasites in vertebrates (Votypka
et al.,
2017),
notably the genus
Hepatozoon
Miller, 1908 (Apicomplexa:
Hepatozoidae), which includes species parasitizing
animals ranging from anurans to mammals (Smith,
1996). Currently, 48 species of
Hepatozoon
are known
from anurans distributed worldwide (Netherlands
et al.
,
2018; Úngari
et al
., 2021).
Brazil has the highest richness of anurans in the world,
totaling approximately 1,180 species, distributed across
all biomes of the country (AMPHIBIAWEB, 2024;
Frost, 2024). However, only four species of hepatozoids
parasitizing anurans are known, mostly occurring in
hosts of the family Leptodactylidae (see Costa
et al.
,
1973; Úngari
et al.,
2021). Te hepatozoids parasitizing
Brazilian anurans are restricted to six host species, located
in areas of the Cerrado and Pantanal biomes in the
central-western region (Costa
et al.,
1973; Leal
et al.,
2015; Úngari
et al.,
2021) and in the Atlantic Forest in
the southeastern region of the country (Ferreira
et al.,
2020). However, there are also records of unidentifed
species of
Hepatozoon
mostly parasitizing leptodactylids,
with two records of parasitism in anurans of the family
Bufonidae (Ferreira
et al
., 2020; Úngari
et al.,
2022).
Te scarce records of
Hepatozoon
demonstrate the lack of
knowledge about parasitism by
Hepatozoon
in Brazilian
anurans, especially in areas within the morphoclimatic
domain of the Caatinga. Tus, the aim of this study was
to register for the frst time parasitism by
Hepatozoon
in two species of bufonid anurans in Caatinga areas in
Brazil, along with the morphometry of the parasites and
parasitized erythrocytes.
MATERIAL AND METHODS
In January and February 2019, three individuals of
Rhinella diptycha
(Cope, 1862) and three of
R. granulosa
(Spix, 1824) were collected from an anthropized
fragment of shrubby Caatinga in the Senador Nilo
Coelho Irrigation Project - Nucleus 01 (9°20’4.68” S;
40°35’11.25” W), in the municipality of Petrolina, state
of Pernambuco, Brazil. Tese anurans were collected
through an active search, using a dip net and talc-free
nitrile gloves, during both the nocturnal and the diurnal
period, near intermittent and perennial water bodies.
After collection, these animals were s0ent to the Animal
Physiology Laboratory of the Universidade Federal do
Vale do São Franciso (UNIVASF), Brazil and at the end
of the studies, they were deposited in the Herpetological
Collection of the Museum of Caatinga Fauna (Coleção
Herpetológica do Museu de Fauna da Caatinga) under
the numbers MFCH 5288-5292 and 5295), which is
located in the Conservation and Management Center
for Caatinga Fauna (Centro de Conservação e Manejo de
Fauna da Caatinga, CEMAFAUNA), at UNIVASF.
To prepare blood smears, samples of blood were collected
via intracardiac puncture or from the ventral abdominal
vein in animals that had previously been euthanized
with an overdose of 10% benzocaine applied topically.
After preparation, the smears were air-dried at room
temperature, fxed with absolute methanol for 10
minutes, and stained with 10% Giemsa for 10 minutes.
Smear analysis was conducted under a light microscope at
a magnifcation of 1000x, with an attached photographic
camera. Te prevalence of parasitism was estimated as the
percentage of infected individuals for each host species,
and the mean intensity of parasitism was calculated based
on the number of parasites found per 2,000 erythrocytes
(Garrido & Pérez-Mellado, 2013). Morphometric
analysis was performed using the ToupView© software,
from measurements that included the length, width
and area of the parasite, the length, width and area of
the parasite nucleus, the length and width of parasitized
erythrocytes, and the length and width of their nuclei,
with data expressed as the mean and standard deviation
(Lima
et al.
, 2021) and range of morphometric variation.
Ethical approval:
Research permit was provided by
Instituto Chico Mendes de Conservação da Biodiversidade
(ICMBio)/Sistema de Autorização e Informação em
Biodiversidade (SISBIO) (Processes n. 29558-1). Te
collection of specimens of
R. diptycha
and
R. granulosa
was and approved by the ethics committee on the use
of animals of the Universidade Federal do Vale do São
Francisco (Processes n. 0001/221018).
RESULTS
Te only parasitic forms found in the blood of both anuran
species were intraerythrocytic gametocytes of
Hepatozoon
spp., parasitizing only hypertrophied erythrocytes. Tree
distinct morphotypes of
Hepatozoon
were found, one
in each host species (Table 1, Fig. 1). In
R. diptycha
,
morphotypes of mature and immature gametocytes were
96
Carvalho
et al.
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
found, with a prevalence of 100% and a mean intensity
of 3.3 ± 3.6, ranging from 2 to 9 parasitized erythrocytes
per 2,000 analyzed. In
R. granulosa
, distinct immature
gametocytes were found, compared with those of
R.
diptycha
, with a prevalence of 66% and mean intensity
of 2.0 ± 2.1, ranging from 1 to 4 parasitized erythrocytes
per 2,000 analyzed.
Te mature gametocytes of
Hepatozoon
sp. from
R.
diptycha
were found to have a cylindrical body with
curved ends in a C or O shape, with a mean length of 9.3
± 1.1 µm and a mean width of 4.7 ± 0.9 µm (n = 11) (Fig.
1A-D). In some of these mature gametocytes analysed,
it was possible to observe the cell body bent inside the
parasitophorous vacuole (Figs. 1A, D). Te nucleus had
the same cellular body shape, with a size of 6.4 ± 0.8 µm
× 2 ± 0.5 µm (Fig 1A). Te gametocytes displaced the
erythrocyte nucleus to the periphery of the cell (Figs. 1A-
D). Te parasitized erythrocytes had a size of 19.0 ± 1.6
µm × 13.6 ± 0.6 µm, and their nuclei were 5.4 ± 0.7 µm
× 6.3 ± 0.5 µm.
Te immature gametocytes had a more robust shape,
wider cell body, granular formations in the cytoplasm
and very subtle curvatures at the ends (Figs. 1E, F),
measuring 8.9 ± 0.8 µm × 4 ± 0.4 µm. Te nuclei had
a square shape with highly condensed chromatin (Fig.
1E), and measured 3.9 ± 0.4 µm × 3.8 ± 1.1 µm, with
less basophilic staining compared with the nucleus of
the parasitized cell (Fig. 1F). Additionally, the immature
gamont also altered the morphology of the erythrocyte,
measuring 16.7 ± 0.1 µm × 13.4 ± 1.7 µm, and pushing
the nucleus of 5.1 ± 0.2 µm × 6.1 ± 0.9 µm to the
periphery of the cell (Fig. 1E, F).
In
R. granulosa
, the immature gametocytes of
Hepatozoon
sp. were found to have a cylindrical shape with subtle
curvatures at the ends, taking on a C shape, with
dimensions of 9.5 ± 0.7 × 4 ± 0.4 µm (n = 8) (Fig. 1G).
Some gametocytes were observed inside parasitophorous
vacuoles (Figs. 1G-L). Te parasite nucleus (4.0 ± 1.0
× 3.7 ± 0.8 µm) presented basophilic staining but with
lower intensity of staining than the erythrocyte nucleus
(Fig. 1L). Additionally, the chromatin showed various
levels of compaction (Figs. 1G-L). Te gametocytes
were capable of displacing the erythrocyte nucleus to
the periphery of the cell (Fig. 1H-J). Te parasitized
erythrocytes measured 18.2 ± 1.2 × 12.4 ± 0.8 µm, and
their nuclei measured 5.4 ± 1.0 × 5.5 ± 0.9 µm.
DISCUSSION
In general, studies on the biodiversity of
Hepatozoon
species in Brazilian anurans can be considered scarce.
Information regarding this genus parasitizing this group
of hosts is concentrated in a few published studies, and
most of them reported unidentifed species (see Table 1).
In Brazil, over the past two decades, with the introduction
of molecular tools combined with morphological studies
on blood and tissue stages, the panorama of
Hepatozoon
biodiversity in anurans has been changing (see Ferreira
et
al.,
2020; Leal
et al.,
2015; Úngari
et al.,
2021, 2022).
For many decades, the parasitism of Brazilian anurans
was associated solely with
H. leptodactyli
(Lesage, 1908)
(Carini, 1908; Pinto, 1925; Cunha & Muniz, 1927;
Pessôa, 1970; Costa
et al
., 1973) or with unidentifed
species of this genus (Table 1). However, recently,
three new species of
Hepatozoon
have been discovered
in Brazilian anurans, based on morphological studies
of blood and tissue stages corroborated by molecular
analyses (Úngari
et al
., 2021). Tis highlights how
underestimated the biodiversity of
Hepatozoon
and other
hemoparasites in Brazilian anurans is.
Te majority of records and descriptions of
Hepatozoon
in Brazil have been made from parasites found exclusively
in fve species of anurans belonging to the genus
Leptodactylus
Fitzinger, 1826 (Leptodactylidae) in areas of
the Cerrado, Pantanal and Atlantic Forest biomes (Carini,
1908; Pinto, 1925; Cunha & Muniz, 1927; Costa
et al.,
1973; Leal
et al.,
2015; Ferreira
et al.,
2020; Úngari
et
al.,
2021, 2022) (Table 1). Te only two records of these
parasites in Bufonidae in this country are in
R. diptycha
from the Atlantic Forest (Ferreira
et al
., 2020) and in
R. mirandaribeiroi
(Gallardo, 1965) parasitizing by
H.
latrensis
in the Pantanal (Úngari
et al.,
2022) (Table 2).
Te mature and immature gametocytes of
Hepatozoon
sp. parasitizing
R. diptycha
were compared in terms of
their morphology and morphometry with other species
previously described in Brazil. Unlike
Hepatozoon
sp.
from
R. diptycha
in the present study,
H. leptodactyli
possesses intraerythrocytic forms apparently without
the parasitophorous vacuole, and the cell body does not
appear to be bent, but has curvature at both ends (Costa
et al
., 1973). Te mature erythrocytes of
H. longinucleus
Úngari, Netherlands, Silva & O’Dwyer, 2021, difer
morphometrically from
Hepatozoon
sp. in
R. diptycha
through having thinner elongated cell bodies and nuclei
(Table 1), in addition to having a parasitophorous
vacuole that takes on the shape of the cell body (Úngari
et al
., 2021). Te mature gametocytes of
H. formosus
Úngari, Netherlands, Silva & O’Dwyer, 2021, have
97
Hepatozoon
of anurans from Caatinga
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
Figure 1
. Intraerythrocytic gametocytes of
Hepatozoon
sp. in the blood smears.
Rhinella diptycha
(A-F). A-D. Mature
gamonts; E-F. Immature gamonts with vacuolization in the parasitic body.
Rhinella granulosa
. G-L. Immature gamonts.
unequal ends of the cell body, with one end thicker than
the other, thus taking on a cone shape, which also shows
subtle curvature (see Úngari
et al.,
2021), while the
gametocytes of
Hepatozoon
sp. from
R. diptycha
have a
curved cell body, larger than the vacuole, and may have a
C or O shape (Figs. 1A-D).
Hepatozoon latrensis
Úngari,
Netherlands, Silva & O’Dwyer, 2021, possess mature
gametocytes without the curvature of the cell body inside
the parasitophorous vacuole (see Úngari
et al.,
2021).
Regarding the immature gametocytes in
Hepatozoon
sp.
from
R. diptycha
, these gametocytes are morphologically
diferent from the mature gametocytes (Figs. 1E-
F). In the immature stage, the parasite nucleus is
centrally positioned, dividing the cytoplasm into two
seemingly equal parts, and it possesses vacuolization in
its cytoplasm (Fig. 1E). Among Brazilian species, there
are morphological descriptions of immature stages of
two species:
H. latrensis
and
H. formosus
(Úngari
et al.,
2021, 2022). Te morphotype that parasitizes
R. diptycha
difers from the immature morphotypes of
H. latrensis
and
H. formosus
, as these have immature gametocytes
similar to the mature ones but with morphometric
diferences (Table 1) (Úngari
et al.,
2021, 2022). In these
two species, in the immature stages, the cytoplasmic area
of the parasite is small and almost imperceptible, with
98
Carvalho
et al.
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
Table 1.
Morphometrics values of intraerythrocytic mature and immature gametocytes of
Hepatozoon
spp. in anurans from Brazil.
Hepatozoon
species
Parasite’s
developmental
stage
Morphometrics
Host speciesReference
Length (µm)
Width
(µm)
Parasite area
(µm²)
Length of
nucleus (µm)
Width of
nucleus
(µm)
Nucleus area
(µm²)
H. formosus
Mature
11.93 ± 0.61
(9.82–13.67)
4.18 ± 0.24
(3.93–4.67)
40.20 ± 3.01
(37.76–44)
5.77 ± 0.23
(5.57–5.97)
3.91 ± 0.23
(3.33–3.93)
17.29 ± 1.29
(15.26–
18.77)
Leptodactylus
labyrinthicus
Úngari
et al.
(2021)
Immature
9.33 ± 0.16
(9.17–9.88)
3.43 ± 0.11
(3.15–3.83)
21.27 ± 0.74
(20.33–22.38)
5.4±1.35
(4.79–6.07)
3.53 ± 0.56
(3.15–3.83)
16.6 ± 0.19
(14.15–
17.16)
L. labyrinthicus
Úngari
et al
.
(2021)
H. longinucleus
Mature
12.58 ± 0.62
(11.5–13.27)
4.17 ± 0.41
(2.4–4.36)
42.24 ± 5.94
(31.37–43.35)
9.21 ± 0.32
(7.5–10.74)
2.18 ± 0.54
(1.71–2.94)
19.16 ± 1.04
(16.78–
20.98)
L. labyrinthicus
Úngari
et al.
(2021)
H. latrensis
Mature
9.88 ± 0.56
(9.06–10.74)
4.35 ± 0.41
(3.27–5.02)
37.46 ± 3
(34.91–43.95)
4.17 ± 0.57
(3.13–5.02)
3.37 ± 0.6
(2.67–4.02)
8.87 ± 1.54
(7.02–9.13)
L. latrans
Úngari
et al.
(2021)
Mature12.28 ± 0.74 ± 0.152.37 ± 0.7
5.65 ± 0.334 ± 0.115.05 ± 0.32
Rhinella
mirandaribeiroi
Úngari
et al.
(2022)
Mature
13.1 ± 0.274.39 ± 0.3452.42 ± 3.255.84 ± 0.53.60 ± 0.3516.30 ± 1.65
L. labyrinthicus
Úngari
et al.
(2022)
Mature
11.63 ± 0.814.63 ± 0.4945.97 ± 4.953.9 ± 0.543.48 ± 0.9310.19 ± 1.92
Leptodactylus
sp.
Úngari
et al.
(2022)
Mature
13.09 ± 0.754.12 ± 0.3947.55 ± 4.55.87 ± 0.845.92 ± 9.4615.84 ± 3.68
L. labyrinthicus
Úngari
et al
.
(2022)
Immature
9.05 ± 0.82.86 ± 0.3316.92 ± 2.94.12 ± 0.372.83 ± 0.318.63 ± 0.60
L. labyrinthicus
Úngari
et al
.
(2022)
H. leptodactyli
Unknow12–16 4–6----
L. latrans
Carini (1908)
Mature 5.1–212.3–8.3-4.63-
L. latrans,
L. pentadactylus
Costa
et al.
(1973)
(Continued Table 1)
99
Hepatozoon
of anurans from Caatinga
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
the diference that in
H. formosus
, the nuclei of immature
gametocytes are larger in relation to the total area of the
parasite (Table 1) (Úngari
et al.,
2021, 2022).
In
Hepatozoon
sp. parasites of
R. granulosa
from
the present study, only immature gametocytes were
found. Tese were characterized by flling the entire
parasitophorous vacuole, with the nucleus following
the shape of the parasite, but without marking the cell
body (Fig. 1G-L). As mentioned earlier, in Brazil, only
H. latrensis
and
H. formosus
have been shown to have
immature gametocytes (see Úngari et al., 2021, 2022). In
some immature gametocytes in
R. granulosa
, the nucleus
appeared segmented (Fig. 1L), and was also characterized
by having a C shape (Fig. 1G-L). Tis was more evident
than in the immature stages of
H. latrensis
and
H.
formosus
(see Úngari
et al
., 2021, 2022). Furthermore,
regarding the nuclei, unlike the immature gametocytes
of
R. granulosa
(Figs. 1G-L), nuclear segmentation was
not observed in the immature gametocytes of
R. diptycha
from the present study. Moreover, the vacuolization
observed in the parasitic body in
R. diptycha
(Fig. 1E)
was not seen in
R. granulosa
. Additionally, these two
species had morphometric diferences, compared with
Hepatozoon
sp. from this host species (Table 1).
Te morphotypes of
Hepatozoon
found in the anurans of
this study each had a prevalence of at least 30%. However,
due to the low host sample size (three individuals of each
species), we were unable to comparatively analyse the
ecological data of
Hepatozoon
sp. parasitism from this
study in relation to data on other species of this genus
parasitizing anurans in Brazil. Nevertheless, the fact that
previously unknown morphotypes of
Hepatozoon
were
found in anurans in Caatinga areas leads us to believe
that there is still potential for studying entirely unknown
biodiversity of
Hepatozoon
species in anurans from this
region. Terefore, this study constitutes the frst report of
Hepatozoon
sp. in anurans in the Caatinga biome, as well
as providing the frst record of parasites of this genus in
R. granulosa
in Brazil.
ACKNOWLEDGMENTS
Tis study was fnanced in part by the Coordenação de
Aperfeiçoamento de Pessoal de Nível Superior – Brasil
(CAPES) - Finance Code 001. Carla Adrielle Costa de
Carvalho was supported by a Master fellowship from
Coordenação de Aperfeiçoamento de Pessoal de Nível
Superior - Brasil (CAPES) (processo: 88887.820630/2023-
00) of the Programa de Pós-graduação em Ciências
Hepatozoon
sp.Mature
9.3 ± 1.1
(8.2–10.4)
4.7 ± 0.9
(2.1–5.6)
37.4 ± 8.9
(16.8–48.3)
6.4 ± 0.8
(5.3–7.3)
2.0 ± 0.5
(0.9–2.5)
11.1 ± 1.6
(8.1–13.4)
R. diptycha
Current study
Immature
8.9 ± 0.8
(8.3–9.4)
4 ± 0.4
(3.7–4.3)
32.3 ± 8.8
(26.1–38.5)
3.9 ± 0.4
(3.6–4.2)
3.8 ± 1.1
(3–4.6)
12.8 ± 4.4
(9.7–15.9)
R. diptycha
Current study
Immature
9.5 ± 0.7
(8.7–10.8)
4 ± 0.4
(3.2–4.5)
39 ± 5.4
(30.4–45.4)
4 ± 1
(2–5.3)
3.7 ± 0.8
(2.2–4.6)
14.1 ± 3.8
(6.8–19.4)
R. granulosa
Current study
Unknow
9.7 ± 2.7
(4.9–13.9)
4.5 ± 0.7
(3.2–5.7)
35.05 ± 10
(20.7–50.0)
4.7 ± 1.4
(2.1–8.6)
4.2 ± 0.75
(2.7–5.4)
16.5 ± 5.3
(6.7–27.8)
L. macrosternum
Leal
et al.
(2015)
Unknow
13.95 ± 3.3
(7.1–17.2)
4.6 ± 0.55
(4–5.8)
43.8 ± 6.2
(30.15–49.2)
4.7 ± 1.1
(2.3–6)
4.2 ± 0.5
(3.65–5.3)
15.3 ± 3.9
(9.5–21.05)
L. podicipinus
Leal
et al.
(2015)
(Continued Table 1)
100
Carvalho
et al.
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
Table 2.
Hepatozoon
spp. in anurans from Brazil, locations and Biomes.
Hepatozoon
species
Host speciesLocalitiesBiomesReferences
H. formosus
Leptodactylus
labyrinthicus
Nova Xavantina, Mato GrossoPantanal
Úngari
et al.
(2021)
H. latrensis
L. latrans
Nova Xavantina, Mato Grosso Pantanal
Úngari
et al.
(2021)
L. labyrinthicus
Araguaiana and Cocalinho, Mato
Grosso
Cerrado
Úngari
et al.
(2022)
Rhinella mirandaribeiroi
Cocalinho, Mato GrossoCerrado
Úngari
et al.
(2022)
Leptodactylus
sp.
Cocalinho, Mato GrossoCerrado
Úngari
et al.
(2022)
H. longinucleus
L. labyrinthicus
Nova Xavantina, Mato Grosso Pantanal
Úngari
et al.
(2021)
H. leptodactyli
L. latrans
São Paulo, São PauloAtlantic forestCarini (1908)
Rio de JaneiroAtlantic forestCunha & Muniz
(1927)
Rio de JaneiroAtlantic forestPinto (1925)
Rio Bonito, São João de Meriti,
Niterói, Cantagalo and Caxias, Rio
de Janeiro;
Guanabara and Florianópolis, Santa
Catarina
Atlantic forestCosta
et al.
(1973)
L. pentadactylus
Goiania, Goiás CerradoPessoa (1970)
Goiania, Goiás CerradoCosta
et al.
(1973)
Santos, São Paulo;
Salvador, Bahia
Atlantic forestCosta
et al.
(1973)
Hepatozoon
sp.
R. diptycha
Petrolina, PernambucoCaatinga
Current study
Jaboticabal, São PauloAtlantic forest
Ferreira
et al.
(2020)
R. granulosa
Petrolina, PernambucoCaatinga
Current study
L. latrans
Barrinha, São PauloAtlantic forest
Ferreira
et al.
(2020)
L. macrosternum
Corumbá, Mato Grosso do SulPantanal
Leal
et al.
(2015)
L. podicipinus
Corumbá, Mato Grosso do SulPantanal
Leal
et al.
(2015)
101
Hepatozoon
of anurans from Caatinga
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
Veterinárias no Semiárido da Universidade Federal do
Vale do São Francisco (PPGCVS - UNIVASF). Fabiano
M. Vieira was supported by a Research Fellowship from
Fundação de Amparo à Ciência e Tecnologia do Estado
de Pernambuco (FACEPE), Brazil (Process: BFP-0206-
2.13/20).
Author contributions: CRediT (Contributor Roles
Taxonomy)
CACC
= Carla Adrielle Costa de Carvalho
GFN
= Gabriela Felix-Nascimento
FMV
= Fabiano Matos Vieira
LAV
= Lucio André Viana
LBR
= Leonardo Barros Ribeiro
Conceptualization:
CACC, GFN, LAV, LBR
Data curation:
CACC, GFN
Formal Analysis:
CACC, GFN, FMV, LAV
Funding acquisition:
GFN, LBR
Investigation:
CACC, GFN
Methodology:
CACC, GFN
Project administration:
CACC, GFN
Resources:
CACC, GFN
Software:
CACC, GFN, FMV
Supervision:
GFN, LAV, LBR
Validation:
CACC, GFN, FMV, LAV, LBR
Visualization:
CACC, GFN, FMV, LAV, LBR
Writing – original draft:
CACC, GFN
Writing – review & editing:
CACC, GFN, FMV, LAV,
LBR
BIBLIOGRAPHIC REFERENCES
AMPHIBIAWEB. (2024).
AmphibiaWeb [On line].
https://amphibiaweb.org
Carini, A. (1908). Sobre a
Haemogregarina leptodactyli
do
Leptodactylus ocellatus
.
Revista da Sociedade Scientifca de São
Paulo, 3
, 59-60.
Costa., S.C.G., Pessoa, S.B., Pereira, N.M., & Colombo, T. (1973). Te life history of
Hepatozoon leptodactyli
(Lesage,
1908) Pessoa, 1970 – a parasite of the common laboratory animal – the frog of the genus
Leptodactylus
.
Memórias
do Instituto Oswaldo Cruz, 71
, 1-18.
Cunha, A.M., & Muniz, J
.
(1927).
Sobre o ciclo endógeno da
Haemogregarina leptodactyli
Lesage, 1908 (
Karyolysus
?).
Memórias do Instituto Oswaldo Cruz, 20
, 307-315.
Ferreira, D.A.R., Perles, L., Machado, R.Z., Prado, C.P.A., & André, M.R. (2020). Molecular detection of Apicomplexan
hemoparasites in anurans from Brazil.
Parasitology Research, 119
, 3469-3471.
Frost, D.R. (2024).
Amphibian Species of the World: An Online Reference Version 6.1
[On line].
https://
amphibiansoftheworld.amnh.org/index.php
Garrido, M., & Pérez-Mellado V. (2013). Prevalence and intensity of blood parasites in insular lizards.
Zoologischer
Anzeiger, 252
, 588-592.
Leal, D.D.M., Dreyer, C.S., Silva, R.J., Ribolla, P.E.M., Paduan, K.S., Bianchi, I., & O’Dwyer, L.H. (2015). Charac-
terization of
Hepatozoon
spp. in
Leptodactylus chaquensis
and
Leptodactylus podicipinus
from two regions of the
Pantanal, state of Mato Grosso do Sul, Brazil.
Parasitology Research, 114,
1541-1549.
Lima, I.G.S., Felix-Nascimento, G., Picelli, A.M., & Ribeiro, L.B. (2021). Contagem diferencial e morfometria de célu-
las sanguíneas nos lagartos
Ameivula ocellifera
(Squamata: Teiidae) e
Tropidurus hispidus
(Squamata: Tropiduridae)
do semiárido brasileiro, com análise dos efeitos por hemoparasitos.
Cuadernos de Herpetología, 35
, 109-119.
Netherlands, E.C., Cook, C.A., & Du Preez, L.H. (2018). Monophyly of the species of
Hepatozoon
(Adeleorina:
Hepatozoidae) parasiting (African) anurans, with the description of three new species from hyperoliid frogs in
South Africa.
Parasitology, 145,
1039-1050.
Pessôa, S.B. (1970). Formas evolutivas do
Hepatozoon leptodactyli
(Lesage, 1908) na sanguessuga
Haementeria lutzi
Pinto, 1920.
Revista Goiana de Medicina, 16
, 35-39.
102
Carvalho
et al.
Neotropical Helminthology, Vol. 18, N
º
1, jan - jun 2024
Pinto, C. (1925). Protozoários observados no Brasil.
Memórias do Instituto Oswaldo Cruz, 18
, 211-302.
Smith, T.G. (1996). Te genus
Hepatozoon
(Apicomplexa: Adeleina).
Journal of Parasitology, 82,
565-585.
Úngari, L.P., Netherlands, E.C., Santos, A.L.Q., Alcantara, E.P., Emmerich, E., Silva, R.J., & O’Dwyer, L.H. 2021.
New insights on the diversity of Brazilian anuran blood parasites: With the description of three new species
of
Hepatozoon
(Apicomplexa: Hepatozoidae) from Leptodactylidae anurans.
International Journal Parasitology:
Parasites and Wildlife, 14
, 190-201.
Úngari, L.P., Netherlands, E.C., Santos, A.L.Q., Alcantara, E.P., Emmerich, E., Silva, R.J., & O’Dwyer, L.H. (2022).
Diversity of Haemogregarine Parasites Infecting Brazilian Anurans, with a Description of New Species of
Dactylosoma
(Apicomplexa: Adeleorina: Dactylosomatidae).
Acta Parasitologica, 67
, 1740-1755.
Votypka, J., Modry, D., Obornık, M., Slapeta, J., & Lukes, J. (2017). Apicomplexa. In: Archibald, J.M., Simpson,
A.G.B., & Slamovits, C. (eds.).
Handbook of the Protists (Second Edition). Springer, Cham
. pp. 567–624.
Received May 5, 2024.
Accepted May 28, 2024.