DOI: https://doi.org/10.52973/rcfcv-luz314.art2
Received: 26/04/2021 Accepted: 02/07/2021
137
Revista Cientica, FCV-LUZ / Vol. XXXI, N°4, 137 - 139, 2021
ABSTRACT
A male canine, two years old, with no known breed, with a history
of diarrhea and ticks, was submitted to a private veterinary clinic in
the City of Guarulhos, São Paulo State, Brazil: presenting weight
loss, alopecia and intense pruritus. The owner reported that the
dog was a native of the City of Votuporanga, São Paulo State.
Blood samples were collected for blood counts and biochemical
measurements, and rapid tests were performed with commercial
kits for the detection of Ehrlichia canis, Anaplasma spp., Borrelia
burgdorferi and Canine distemper. Due to the hematological and
biochemical changes, the negative results, and the origin of the
animal, it was decided to perform a rapid test for leishmaniasis,
where this test presented a positive result, later confirmed
by Polymerase Chain Reaction. The Leishmaniose continue
advancing, being able to settle in non-endemic areas.
Key words: Canine; metropolis; Leishmania (L) infantum;
Lutzomyia longipalpis
RESUMEN
Un canino macho, de dos años, mestizo, con antecedente de
diarrea y garrapatas, con adelgazamiento, alopecia y prurito
intenso, fue remitido a una clínica veterinaria privada de la ciudad
de Guarulhos, estado de São Paulo, Brasil. El tutor informó que el
perro era de la ciudad de Votuporanga, estado de São Paulo. Se
recolectaron muestras de sangre para hemograma y mediciones
bioquímicas, y se realizaron pruebas rápidas con kits comerciales
para la detección de Ehrlichia canis, Anaplasma spp., Borrelia
burgdorferi y moquillo canino. Ante los cambios hematológicos y
bioquímicos, los resultados negativos y el origen del animal, se
decidió realizar una prueba rápida para leishmaniasis, donde se
obtuvo un resultado positivo, posteriormente conrmado por una
prueba de Reacción en Cadena de la Polimerasa. La leishmaniosis
continúa avanzando y puede establecerse en áreas no endémicas.
Palabras clave: Canino; metrópoli; Leishmania (L) infantum;
Lutzomyia longipalpis
A Case Non-Autochthonous Canine Visceral Leismaniosis in the
Municipality of Guarulhos, São Paulo, Brazil
Un Caso de Leismaniosis Visceral Canina No Autóctona en el Municipio de Guarulhos,
São Paulo, Brasil
Polliana Pimentel
1
, Leandro Macedo-Miranda
2
, Katia Klemba
1
, Daniele Silva-Roque
3
, Paulo Usignolo
Carnauba-Vicente
3
, João Carlos Lopes-Costa
2
, Nathálya dos Santos-Martins
4
and Diego Carvalho-Viana
2,5
*
1
PUC. São Paulo, Brasil.
2
Programa de Pós-Graduação em Ciência Animal, Universidade Estadual do Maranhão. São Luís, Maranhão,
Brasil.
3
Universidade Guarulhos. São Paulo, Brasil.
4
Universidade Federal do Maranhão (UFMA). São Luís, Maranhão, Brasil
5
Universidade Estadual da Região Tocantina do Maranhão (UEMASUL). Imperatriz, Maranhão, Brasil.
*Email: dieob@bol.com.br
Un Caso de Leismaniosis / Pimentel et al.___________________________________________________________________________
138
INTRODUCTION
Leishmaniose is a chronic infectious, cosmopolitan and zoonotic
disease caused by protozoa of the family Trypanosomatidae,
genus Leishmania [9]. The species responsible for causing Canine
Visceral Leishmaniasis (CVL) was Leishmania (L) infantum [6]. In
Brazil, the main vector involved in transmission is the phlebotomine
Lutzomyia longipalpis [1], with dogs (Canis lupus) being the main
reservoirs in the urban environment, as they can present a large
amount of amastigote forms in the skin, increasing the chances
of infecting the vector [2].
CVL may present in symptomatic and asymptomatic forms,
with the symptomatic form having several clinical signs, such as
poor body condition, lymphadenomegaly, cutaneous alterations,
onychogrifosis, purulent keratoconjunctivitis, epistaxis, mucous
pallor, among others [3]. According to the Visceral Leishmaniasis
Surveillance and Control Program in the State of São Paulo, the
L. longipalpis vector was detected in 177 Municipalities, being
present in 76 Municipalities where there was canine and human
transmission and, according to the same, the Municipality of
Guarulhos [2], which was classied as "silent" (without conrmation
of human and / or canine cases), not receptive (there is no
conrmation of presence of the vector). Therefore, the objective
of this work was to report the rst case of non-autochthonous CVL
in the Municipality of Guarulhos.
MATERIALS AND METHODS
A male canine, two years (y) old was selected to be sent to
a private veterinary clinic in the City of Guarulhos, State of São
Paulo, Brazil, which had been treating a dermatopathy for two
months (mon) with suspicion of hemoparasitosis on July 16, 2018.
The owner reported as main complaints: diarrhea, weight loss,
alopecia and intense itching, and that the animal was not improving
with the use of Cephalexin (15 miligrams (mg)·kilogram
-1
(kg), twice
a day-d-). It was also reported that the dog had infestation by ticks
and that this from Votuporanga City, São Paulo State, about 530
kilometers (km) away from the city which was attended (FIG.1).
Clinical examination revealed generalized alopecia, tachycardia,
tachypnea, cachexia and onychogrifosis. The mucous membranes
were hypocorate, rectal temperature of 39.8 ºC, generalized
lymphadenomegaly and abdominal palpation pain. Blood
samples were then collected for blood counts and biochemical
measurements by jugular vein puncture and then sent to the
laboratory for clinical analysis. A coproparasitological examination
was performed in which the presence of Giardia spp. and skin
scraping where the presence of mites was not detected. Due
to the history of infestation by ticks (Rhipicephalus sanguineus)
and the presence of some clinical and laboratory alterations, a
blood sample was collected for diagnostic kits for the detection of
Ehrlichia canis, Anaplasma spp., Borrelia burgdorferi (Canine Snap
4Dx Plus Test, IDEXX®) and Canine distemper (Alere Cinomose
Ag Test Kit®) in which both presented negative results.
RESULTS AND DISCUSION
Considering the negative results and the animal’s history, which
came from the Municipality of Votuporanga, the chosen test to
perform a rapid test for leishmaniasis (Alere Leishmaniose Ac
Test Kit®), which consists of a chromatographic immunoassay
(CI) [8], in which the qualitative detection of antibodies was
carried out by means of the association of the antigens rK39,
rK26 and rK9, where it showed a positive result. As a result, the
blood sample was evaluated by the Polymerase Chain Reaction
(PCR), test to detect the deoxyribonucleic acid (DNA) of the agent
using the Wizard Genomic DNA Purication Kit, Promega® for
extraction and purication of DNA, and primers RVI (sense;5’-
CTTTTCTGGTCCCGCGGGTAG-3’) and RV [6] (antisense;5’-
CCACCTGGCCTATTTTACACCA-3’) to amplify the target sequence
of the L. infantum DNA fragment on August 1, 2018. The PCR
result was positive and, after conrming the result, a joint decision
was made with the animal’s owner, and the euthanasia procedure
was approved according to resolution 1,000 of May 11, 2012 [1].
Because it is a disease with a wide variety of clinical signs, the
diagnosis can become confused, since it can be confused with
FIGURE 1. Map of the state of São Paulo highlighting the municipalities
related to occurrence of the case of leishmaniasis. The following
municipalities are cited: Guarulhos, the rst case of non-autochthonous
LVC; Votuporanga, origin of the animal; and São Paulo
___________________________________________________________Revista Cientica, FCV-LUZ / Vol. XXXI, N°4, 137 - 139, 2021
139
other pathologies [4], which can take advantage of failures in the
immune system triggered by the disease.
The biochemical parameters altered in the blood test were:
increased bilirubin (0.81 mg·deciliter
-1
-dL-), alkaline phosphatase
(286 international units per litre -L-), urea (237.1 mg·dL
-1
) and
creatinine (1.72 mg·dL
-1
), demonstrating a hepatic and renal
impairment. As for hematological parameters, normocytic,
normocromic and non-regenerative anemia were observed, in
addition to thrombocytopenia. These ndings were similar to those
found by Torrecilha et al. [9] in which they studied the correlation
between peripheral parasitic load and clinical and laboratory
abnormalities in dogs with CVL, where they found alterations
in the hepatic and renal parameters of the dogs studied, and
non-regenerative anemia.
There are several techniques for the diagnosis of CVL, such
that the association between them provides a greater accuracy,
since there is no technique that alone can give a totally accurate
diagnosis of the disease [5]. The diagnosis was supported by
clinical and pathological ndings and validated by serological and
molecular techniques [4]. The rapid test for leishmaniasis (Alere
Leishmaniose Ac Test Kit®) is a good alternative for screening
patients, since it has a good specicity, however its sensitivity is
variable, being ideal to combine it with a more sensitive technique
like PCR.
As described previously, the animal came from the Municipality
of Votuporanga, located in the Northwest Region of the São Paulo
State, about 535 km from Guarulhos (FIG. 1). In this Region there is
a wide distribution of the L. longipalpis vector, and the Municipality
of Votuporanga was classied as an area of transmission of human
visceral leishmaniasis (HVL) and CVL [2].
Studies have shown that HVL in the São Paulo State has
expanded over the (y) in the Northwest-Southeast direction towards
the Bauru Region, as a reection of the migration of workers from
other endemic regions of the Country. This expansion also had
as determinant factors the adaptation of the vector to the climatic
conditions, the migration of dogs from endemic to non-endemic
areas, as well as failures in vector control actions in the domestic
reservoirs [6, 7]. Another factor to consider is the possibility of
other vectors acting on the CVL and HVL transmission [3], in which
it was pointed out the sandy Pintomyia scheri as a potential
vector of L. infantum in the region of São Paulo, since this one
showed susceptibility to parasite development, although it has a
lower transmission capacity than L. longipalpis.
CONCLUSIONS
The increasing expansion of Leishmaniose in the São Paulo
State, the circulation of people and animals aected from endemic
to non-endemic areas, as well as factors favoring the presence
of the vector in the urban environment, such as deforestation,
presence of garbage and lack of sanitation, and as the possibility
of other vectors acting in the transmission of the disease, are
conditions for the leishmaniose to continue advancing, being
able to settle in non-endemic areas. State surveillance systems
are important in the sense of creating a network of information
about the quality of information and they are possible to generate
a connection between neighboring States to provide a strategic
government management to combat anthropozoonotic diseases.
ACKNOWLEDGEMENTS
Thank Coordination for the Improvement of Higher Education
Personnel (CAPES) and Maranhão Foundation for the Support of
Scientic and Technological Research and Development (FAPEMA),
Brazil.
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