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Comp Clin Pathol (2010) 19:383–387

DOI 10.1007/s00580-010-1041-2

BRIEF COMMUNICATION

Rangelia vitalli in dogs in southern Brazil


Raqueli Teresinha França & Aleksandro Schafer Da Silva & Francine Chimelo Paim &
Marcio Machado Costa & João Fabio Soares & Cinthia Melazzo Mazzanti &
Sonia Terezinha dos Anjos Lopes

Received: 21 March 2010 / Accepted: 26 May 2010 / Published online: 8 June 2010
# Springer-Verlag London Limited 2010

Abstract This article aims to describe seven cases of dogs short communication published by Antonio Carini about
naturally infected with Rangelia vitalli. Clinical findings, the most common infectious and parasitic diseases of
hematological parameters, parasitological diagnosis, and domestic animals in Brazil at that time, the observation of
treatments were evaluated. On physical examination, these a disease in dogs called “nambiuvú” was mentioned (Carini
animals showed pale mucous membranes, hyperthermia, 1908). Later, another author published two scientific papers
apathy, and blood dribbling down from the ear margins. R. characterizing the morphology of this unusual protozoan
vitalli merozoites were observed inside of erythrocytes, parasite based on observations made under a light micro-
neutrophils, and macrophages in blood smear. Important scope and describing the epidemiological, clinical, and
hematological findings observed in these cases were severe pathological aspects of the disease caused by this atypical
anemia and thrombocytopenia. The animals were treated with piroplasm (Pestana 1910a, b). In 1914, Carini and Maciel
therapeutic protocol based on prednisone, doxycycline, and (1914) wrote an article about this disease proposing that the
dipropionate which had great curative efficacy to rangeliosis. name of this previously unidentified canine piroplasm
should be changed to R. vitalli to honor the investigator
Keywords Rangeliosis . Nambiuvú . Canine . Anemia (Pestana 1910a, b), who first observed the presence of this
organism within endothelial cells and red blood cells of
Brazilian dogs affected by the protozoa R. vitalli.
Introduction This parasite usually affects young dogs from rural and
periurban areas during the hot season, when the number of
Rangelia vitalli is a member of the protozoan phylum ticks is greater, but can also occur during the year in adult
Apicomplexa, order Piroplasmorida (Loretti et al. 2003; animals. The disease is characterized by anemia, jaundice,
Spagnol et al. 2003), that causes a tick-borne disease fever, splenomegaly, generalized lymph node enlargement,
referred to as “nambiuvú” (=blood dribbling down from the hemorrhage along the gastrointestinal tract, and persistent
ear margins) or “peste de sangue” (=bleeding plague). The bleeding through the tips of the pinnae, external surface of
original reference to this disease comes from 1908. In a the ears, nose, and oral cavity (Pestana 1910a, b; Loretti
and Barros 2005). According to the literature, ticks
Amblyomma aureolatum and Rhipicephalus sanguineus
R. T. França (*) : A. S. Da Silva : F. C. Paim : M. M. Costa : serve as vectors of R. vitalli, and wild carnivores and birds
C. M. Mazzanti : S. T. dos Anjos Lopes (that are resistant to the infection by this protozoa) play an
Department of Small Animals,
important role as reservoirs of the pathogen in rural areas
Universidade Federal de Santa Maria,
Santa Maria, Rio Grande do Sul, Brazil (Loretti and Barros 2004).
e-mail: raquelifranca@yahoo.com.br Tentative clinical diagnosis of R. vitalli infection is
based on the history of the animal, clinical signs, hemo-
J. F. Soares
gram, and favorable response to therapy (Loretti and
Department of Preventive Veterinary Medicine and Animal
Health, Universidade de São Paulo, Barros 2004). Definitive diagnosis has been carried out
São Paulo, São Paulo, Brazil through microscopical examination of smears from tissue
384 Comp Clin Pathol (2010) 19:383–387

samples collected during necropsies or histological sec- Results


tions. However, molecular phylogenetic analysis is neces-
sary to better characterize this parasite and clarify its The initial diagnosis of R. vitalli was based on the
taxonomic status. morphological analysis of piroplasm in blood smears
In the literature, we have a few studies about the (Fig. 1). In the blood of five dogs, intracellular organisms
protozoan R. vitalli, likely due to the lack of knowledge inside macrophages (monocytes—Fig. 1b), erythrocytes
about the parasite and/or merely because the protozoan was (Fig. 1c), and neutrophils (Fig. 1d) were observed. In the
diagnosed only in the south of Brazil. Therefore, this article other two animals, clinical diagnosis was performed,
aims to describe seven cases of dogs naturally infected with followed by drug therapy.
R. vitalli. In infected dogs, there was a significant reduction in
erythrocyte count, hemoglobin, and hematocrit when
compared to reference values (Feldman et al. 2000). Most
Materials and methods affected animals in this study (dogs 1, 3, 4, 5, and 6) had a
mean of increased MCV and mean of decreased MCHC,
Seven affected dogs referred to a local University Veteri- characterizing macrocytic–hipochromic anemia. Dog 2 had
nary Hospital (UVH-UFSM, Santa Maria, RS, Brazil) were macrocytic–normochromic anemia. For dog 7, the mean of
included in this study. Among the animals, four were males MCV and the mean of MCHC did not differ from reference
and three were females, ranging from 6 months to 5 years values, characterizing normocytic–normochromic anemia
of age. Two dogs were mongrel (dogs 1 and 2) and the (Table 1). The presence of anisocytosis and polychromasia
others were of the following breeds: Border Collie (dog 3), were observed in seven dogs.
Cimarron (dog 4), Collie (dog 5), Boxer (dog 6), and Dog 4 showed leukocytosis as a consequence of
Cocker Spaniel (dog 7). Clinical findings, hematological neutrophilia and lymphocytosis. In other animals, the
parameters, parasitological diagnosis, and treatment were number of total leukocytes did not change. TPP levels
analyzed. increased in dogs 2 and 4 and decreased in dog 5 when
On physical examination, these animals showed pale compared to reference values. The number of platelets
mucous membranes, hyperthermia, apathy, and blood decreased dramatically in all affected dogs (Table 1).
dribbling down from the ear margins (Fig. 1a). Ticks were The therapy used to combat the parasite R. vitalli was
not found in five animals, but the owners reported having seen effective for six dogs. These animals showed clinical
ticks in the last 7 days on these dogs. In two of the dogs, ticks improvement, and hematological parameters returned to
of the species R. sanguineus were found. Two milliliters of normal levels. Dog 2 was the only one who did not respond
blood from all dogs was collected for further evaluation of to therapy and died on the day the diagnosis was made.
hematological parameters and kept in tubes containing
ethylenediamine tetraacetic acid. Erythrocytes count, hemat-
ocrit (Ht), hemoglobin concentration, mean corpuscular Discussion
volume (MCV), mean corpuscular hemoglobin concentration
(MCHC), total plasma proteins (TPP), platelet number, and R. vitalli has been referred in the international literature as
total leukocyte count were evaluated. Smears were mounted Babesia vitalli of Brazilian dogs (Wenyon 1926) and
and stained by panoptic and/or Giemsa methods. Ht and considered synonym for Babesia canis (Levine 1973;
hematometric indices were evaluated according to the Peirce 2000). It is postulated by some investigators that
methodology proposed by Feldman et al. (2000). Erythro- the intracellular form of Rangelia found inside the
cytes and leukocytes count, hemoglobin concentration, and endothelial cells is possibly Toxoplasma, and that the tissue
platelet counts were determined using an electronic counter. stage of Rangelia has no connection with blood stage
TPP was carried out by refractometry. which, according to them, is in reality B. canis (Wenyon
From the samples collected, blood smears were per- 1926; Moreira 1938; Paraense and Vianna 1948). It is stated
formed according to Pestana (1910a, b). Each slide was in one publication that the masses of R. vitalli merozoites
mounted with blood collected from the cephalic vein, being described originally within the cytoplasm of the
stained by the panoptic and/or Giemsa methods, and endothelial cells were, in fact, agglomerates of these
visualized at a magnification of ×1,000. organisms accumulated inside the lumen of the capillary
After the diagnosis was established, standard treatment blood vessels (Levine 1973). As shown in the study of
for nambiuvú with corticosteroids (prednisone—2 mg kg−1) Fighera (2007), in only 4% of cases of rangeliosis, the
and doxycycline (5 mg kg−1) was administered orally twice parasites are found in the blood stream. However, in this
a day for 3 days. At the same time, we used a single dose of study, 71.4% of dogs with clinical signs typical of
imidocarb dipropionate (5 mg kg−1, sc). rangeliosis presented the parasite inside blood cells.
Comp Clin Pathol (2010) 19:383–387 385

Fig. 1 Dog infected with R.


vitalli showing pale mucosa
and bleeding at the edges of the
ears (a). Parasites inside macro-
phages in blood smears stained
with Giemsa method (b) and
within erythrocyte (c) and neu-
trophils (d) in panoptic method
smear

The definitive diagnosis of parasitism by R. vitalli is erythrocytes. This reinforces the importance of “blood smear”
problematic since this parasite has been observed in blood as a diagnostic method of this disease in dogs, since this
smears of spontaneous and experimental cases of rangelio- technique has so far not been considered as a method in the
sis (Loretti and Barros 2004). This intracellular parasite has diagnosis of R. vitalli. Our research group is working on a
been found in cytology and histology, endothelial cells of phylogenetic study based on molecular methods such as
blood capillaries, from necropsy samples, cytology of PCR amplification and DNA sequencing to characterize the
lymph nodes and bone marrow, and also is rarely found protozoan and further to develop a specific PCR for R.
within red blood cells and free in the blood (Krauspenhar et vitalli. This study is in progress.
al. 2003; Loretti et al. 2003; Loretti and Barros 2005). Researchers (Krauspenhar et al. 2003; Loretti and Barros
Contrary to what has been reported, we observed that in five 2005) observed evidences that the protozoan induces
of seven dogs with rangeliosis in our study, the protozoan was hemolytic anemia immune-mediated, causing a severe
found inside the macrophages (monocytes), neutrophils, and regenerative anemia together with spherocytosis and eryth-

Table 1 Hematology of dogs naturally infected with Rangelia vitalli

Hemogram and reference values Animals infected whit Rangelia vitalli

Dog 1 Dog 2 Dog 3 Dog 4 Dog 5 Dog 6 Dog 7

Erythrocytes total (×106/µl) (5.5–8.5a) 1.81 2.56 2.57 2.48 1.58 2.6 2.3
Hemoglobin (g/dl) (12–18a) 4.4 7.0 6.1 6.5 3.7 5.9 6.0
Hematocrit (%) (37–55a) 16 23 20 20 14 20 17
MCV (fl) (60–77a) 85.2 89.8 77.8 80.6 88.0 77.9 73.9
MCHC (%) (32–36a) 27.8 33.0 30.5 31.5 26.0 29.5 35.2
TPP (g/dl) (6–8a) 6.3 8.6 6.0 9.6 5.2 8.0 7.0
Platelets (×103/µl) (200–500a) 23 24 34 106 29 27 70
Leukocyte total (×103/µl) (6–17a) 12.3 7.1 10.1 35.3 9.7 10.2 9.2

Hemogram was carried out on the day of clinical and parasitological diagnosis
a
Reference values
386 Comp Clin Pathol (2010) 19:383–387

rophagocytosis, which are the causes of autoimmune clinical features of the disease—bleeding edge of the ears.
anemia (Feldman et al. 2000; McCullough 2003). In our It is important to remember that this therapy is also used as
study, we also observed severe anemia in all dogs (Table 1), a method of diagnosis of this parasite (medical diagnosis).
with significant reduction of total erythrocyte, hematocrit, In this study, six dogs were treated aiming to cure the
and hemoglobin. The cause of anemia was not investigated; disease, except dog 2 that did not respond to treatment and
however, the presence of anisocytosis and polychromasia died. Loretti and Barros (2005) in their study reported
suggests that the dogs had regenerative anemia. spontaneous death of eight affected animals. Three of those
Only dog number 4 showed leukocytosis when evaluated animals showed jaundice and died acutely in approximately
(Table 1). The same animal together with dog 2 had 1 week after the observation of initial clinical signs.
increase in the concentration of TPP, which characterizes an Therapy with doxycycline was started in the late stage of
inflammatory response in order to combat parasitism. the disease in one of the icteric dogs and was unsuccessful.
According to researchers, leukocytosis is due to neutro- One animal that died after a protracted clinical course of 2–
philia with regenerative left shift as a consequence of 3 months had anemia. Other animals had recovery
prolonged stimulation, which may be accentuated and approximately 48 h after therapy with imidocarb dipropi-
nonspecific of the bone marrow (Fighera 2007). In some onate and blood transfusion, and another after treatment
cases, this leukocytosis may be presented in the form of a with doxycycline and glucocorticoid therapy. Three animals
leukemoid reaction. Other common findings include lym- died 24–48 h after therapy with diminazene aceturate. In
phocytosis and monocytosis (Loretti and Barros 2004; our study, we obtained the cure of all animals treated,
Fighera 2007), a case where an increase in total leukocytes which allows us to suggest that therapy with the three drugs
was due to neutrophilia and lymphocytosis. Based on our is the best treatment protocol for rangeliosis.
findings and literature, we conclude that the leukogram is Based on the findings, it is concluded that R. vitalli is
inconsistent in rangeliosis, ranging from leukopenia to pathogenic for dogs, causing severe anemia and thrombo-
leukocytosis or without leukocyte alterations as observed in cytopenia. Blood smear is an important method to visualize
most of our cases reported here. the parasite within red blood cells, neutrophils, and macro-
The pathogenesis of the characteristic clinical sign (ear phages. The therapeutic protocol based on prednisone,
bleeding—nambiuvú) usually observed in cases of dogs doxycycline, and dipropionate imidocarb is effective in the
infected is still unclear (Loretti and Barros 2005). Dissem- treatment of rangeliosis.
inated intravascular coagulation secondary to endothelial
damage caused by the replication of this organism inside
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