113 Endoparasites of anurans in the seven cities national park Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 Neotropical Helminthology Neotropical Helminthology, 2025, vol. 19 (1), 113-119 ORIGINAL ARTICLE / ARTÍCULO ORIGINAL RANGELIA VITALII (CARINI, 1908) (APICOMPLEXA, PIROPLASMORIDA) IN DOMESTIC DOG IN SOUTHERN BRAZIL: A CASE REPORT RANGELIA VITALII (CARINI, 1908) (APICOMPLEXA, PIROPLASMORIDA) EN PERRO DOMÉSTICO EN EL SUR DE BRASIL: REPORTE DE CASO R ANGELIA VITALII (CARINI, 1908) (APICOMPLEXA, PIROPLASMORIDA) EM CANINO DOMÉSTICO NO SUL DO BRASIL: RELATO DE CASO Alexsander Ferraz 1* , Laura Dias Petricione 2 , Eugênia Tavares Barwaldt 2 , Renata Fontes Ongaratto 1 , T aís Cezimbra Reichow 1 , Wesley Porto de Oliveira 1 , Rodrigo Leite dos Santos 1 , Camila Moura de Lima 2 , Fábio Raphael Pascoti Bruhm 1 & Leandro Quintana Nizoli 1 ISSN Versión Impresa 2218-6425 ISSN Versión Electrónica 1995-1403 DOI: https://dx.doi.org/10.62429/rnh20251911937 Universidad Nacional Federico Villarreal Volume 19, Number 1 (jan - jun) 2025 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. ABSTRACT Rangeliosis is an extravascular hemolytic disease caused by the piroplasm Rangelia vitalii (Carini, 1908). T is protozoan is transmitted by ticks of the species Amblyomma aureolatum (Pallas, 1772) and infects erythrocytes, endothelial cells, and leukocytes, leading to a variety of clinical signs such as anemia, thrombocytopenia, splenomegaly, hepatomegaly, jaundice, and bleeding from the nose, oral cavity, and ear tips. T is study aims to report a case of rangeliosis in a domestic dog in southern Brazil. A female, adult, mixed-breed dog from a rural area was admitted to a Veterinary Hospital. Clinical examination revealed necrosis of the ear tips with active bleeding and abdominal hematomas. After a few days, the patient also developed jaundice, hematuria, bilirubinuria, subconjunctival hemorrhage, and epistaxis. 1 Federal University of Pelotas, Veterinary School, Department of Preventive Veterinary. Brazil. 2 Federal University of Pelotas, Veterinary School, Department of Veterinary Clinics. Brazil. * Corresponding author: xanderferraz@yahoo.com.brAlexsander Ferraz: https://orcid.org/0000-0002-0424-6249Laura Dias Petricione: https://orcid.org/0000-0003-4731-7577Eugênia Tavares Barwaldt: https://orcid.org/0000-0002-4902-1203Renata Fontes Ongaratto: https://orcid.org/0000-0002-4006-8830T aís Cezimbra Reichow: https://orcid.org/0009-0005-0131-6685Wesley Porto de Oliveira: https://orcid.org/0009-0009-8533-2592Rodrigo Leite dos Santos: https://orcid.org/0009-0003-4631-9025Camila Moura de Lima: https://orcid.org/0000-0001-9306-705XFábio Raphael Pascoti Bruhm: https://orcid.org/0000-0002-4191-965XLeandro Quintana Nizoli: https://orcid.org/0000-0002-0767-4097
114 Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 Ferraz et al. Hematological analysis indicated anemia and thrombocytopenia, while biochemical tests revealed elevated serum urea levels. Te defnitive diagnosis was confrmed through a stained blood smear, which revealed the presence of R. vitalii inside leukocytes and free in plasma. In addition to supportive treatment, doxycycline and imidocarb dipropionate were administered. However, due to disease progression, the patient ultimately succumbed. Canine rangeliosis is likely an underdiagnosed disease; therefore, it is crucial to consider it as a diferential diagnosis in patients presenting compatible clinical manifestations and laboratory fndings, particularly in those from rural or forested areas or with access to such environments. Keywords: bleeding – diagnosis – protozoan – rangeliosis – tick RESUMO Rangeliose é uma doença hemolítica extravascular, causada pelo piroplasma Rangelia vitalli (Carini, 1908). Este protozoário é transmitido através dos carrapatos da espécie Amblyomma aureolatum (Pallas, 1772) e infectam eritrócitos, células endoteliais e leucócitos, ocasionando uma variedade de sinais clínicos, como anemia, trombocitopenia, esplenomegalia, hepatomegalia, icterícia e sangramento pelo nariz, cavidade oral e ponta de orelha. Este trabalho teve como objetivo, relatar um caso de rangeliose em canino doméstico no sul do Brasil. Foi atendido num Hospital Veterinário, um canino, fêmea, adulto, sem raça defnida, proveniente de área rural. No exame clínico, apresentava necrose de ponta de orelhas com sangramento ativo e hematomas no abdômen, e após alguns dias começou a apresentar também, icterícia, hematúria, bilirrubinúria, hemorragia subconjuntival e epistaxe. Anemia e trombocitopenia foram observadas no hemograma, e no exame bioquímico, aumento dos níveis séricos de uréia. O diagnóstico defnitivo foi obtido através do esfregaço sanguíneo corado, onde observou-se a presença de R. vitalii no interior dos leucócitos e livre no plasma. Além do tratamento de suporte, foram utilizadas aplicações de doxiciclina e dipropionato de imidocarb, porém, devido ao agravamento do quadro, o paciente acabou evoluindo para óbito. A rangeliose canina é possivelmente uma doença subdiagnosticada, portanto, é importante que seja considerada como diagnóstico diferencial em pacientes com manifestações clínicas e achados laboratoriais compatíveis, principalmente naqueles procedentes de áreas rurais ou próximas a forestas ou que tenham tido acesso a estes locais. Palavras chave: carrapato – diagnóstico – protozoário – rangeliose – sangramento RESUMEN La rangeliosis es una enfermedad hemolítica extravascular, causada por el piroplasma Rangelia vitalli (Carini, 1908). Este protozoario se transmite a través de garrapatas de la especie Amblyomma aureolatum (Pallas, 1772) e infecta eritrocitos, células endoteliales y leucocitos, provocando una variedad de signos clínicos como anemia, trombocitopenia, esplenomegalia, hepatomegalia, ictericia y hemorragias por la nariz, cavidad oral y punta de las orejas. Este trabajo tuvo como objetivo informar un caso de rangeliosis en un perro doméstico en el sur de Brasil. Fue atendido en un Hospital Veterinario, un perro, hembra, adulta, sin raza defnida, proveniente de una zona rural. En el examen clínico, presentaba necrosis en las puntas de las orejas con sangrado activo y hematomas en el abdomen, y después de algunos días comenzó a presentar también ictericia, hematuria, bilirrubinuria, hemorragia subconjuntival y epistaxis. Se observó anemia y trombocitopenia en el hemograma, y en el examen bioquímico, un aumento de los niveles séricos de urea. El diagnóstico defnitivo se obtuvo a través del frotis sanguíneo teñido, donde se observó la presencia de R. vitalli dentro de los leucocitos y libre en el plasma. Además del tratamiento de soporte, se utilizaron aplicaciones de doxiciclina y dipropionato de imidocarb; sin embargo, debido al empeoramiento de la condición, el paciente terminó falleciendo. La rangeliosis canina es posiblemente una enfermedad subdiagnosticada, por lo tanto, es importante considerarla como diagnóstico diferencial en pacientes con manifestaciones clínicas y hallazgos de laboratorio compatibles, especialmente en aquellos provenientes de zonas rurales o cercanas a bosques, o que hayan tenido acceso a estos lugares. Palabras clave : diagnóstico – garrapata – protozoario – rangeliosis – sangrado
115 Rangelia vitalii in domestic dog in southern Brazil Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 INTRODUCTION Rangelia vitalli (Carini, 1908) is a protozoan belonging to the Phylum Apicomplexa and Order Piroplasmorida, which infects canids. It is transmitted by ticks of the species Amblyomma aureolatum (Pallas, 1772), leading to extravascular hemolysis (Soares et al. , 2018). Rangelia vitalii infects erythrocytes, endothelial cells, and leukocytes, causing a range of clinical signs such as anemia, thrombocytopenia, hemorrhage, fever, splenomegaly, hepatomegaly, lymphadenopathy, jaundice, and bleeding from the nose, oral cavity, and ear tips (Silva et al ., 2011; Soares et al ., 2011; Rodrigues et al ., 2022). While observing clinical signs is important, they are nonspecifc and common to other hemoparasitic diseases. Terefore, the defnitive diagnosis of canine rangeliosis involves a combination of clinical, laboratory, and imaging methods. Although clinical symptoms are crucial, confrmation requires specifc laboratory tests. Serological tests, such as indirect immunofuorescence and polymerase chain reaction (PCR) (Soares et al. , 2011) and observation of parasites in peripheral blood smear (França et al ., 2010) are commonly used to detect the presence of the protozoan in the animal’s blood. Additionally, hematological analysis may reveal characteristic changes, such as hemolytic anemia (Soares et al ., 2015).Treatment of rangeliosis typically involves the administration of antiprotozoal drugs, such as imidocarb dipropionate (Borrás et al ., 2020). Furthermore, supportive therapy is essential to address anemia and other complications, and blood transfusions may be necessary in severe cases (Loretti, 2012). Complete recovery depends on the severity of the infection and the individual response of each animal to treatment.Terefore, the aim of this study was to report a case of rangeliosis in a domestic canine in southern Brazil. MATERIALS AND METHODS A female adult, mixed-breed canine from a rural area was admitted at the Veterinary Hospital of the Federal University of Pelotas; Rio Grande do Sul state, Brazil. In the anamnesis, the owners reported that the animal was apathetic, hypothermic, with ear lesions that were not healing and were bleeding. Upon physical examination, it was noted that the patient was thin, with pale pink mucous membranes, and had fever (39.3ºC). Te clinical exam revealed necrosis at the tips of the ears with active bleeding (Figure 1A) and bruising on the abdomen and around the body (with less intensity) (Figure 1B). A few days after hospitalization, the abdominal hematoma worsened, and the patient began to present with jaundice, hematuria (Figure 1C), bilirubinuria (Figure 1D), subconjunctival hemorrhage (Figure 1E), and epistaxis. Complementary tests including a complete blood count, biochemical analysis, imaging via ultrasonography, and due to the clinical signs observed, a hemoparasite investigation was also requested. Figure 1. Dog with rangeliosis presenting ear tip necrosis (A), abdominal hematomas (B), hematuria (C), bilirubinuria (D) and subconjunctival hemorrhage (E).
116 Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 Ferraz et al. Ethic aspects: Te authors argue that all national and international ethical regulations were met. RESULTS AND DISCUSSION In the complete blood count performed on the day of ad - mission, macrocytic/normochromic anemia and thrombo - cytopenia were observed. Morphological evaluation revea - led anisocytosis, polychromasia, Howell-Jolly bodies, and platelet aggregation. In the biochemical analysis, increased urea levels (110.63 mg/dL) and icteric serum were noted. With the worsening of clinical signs, additional hematolo - gical tests were conducted, which confrmed the progression of the condition (Table 1). Abdominal ultrasonography re - vealed splenomegaly with enlargement of the splenic lym - ph nodes. Parasitized cells are recognized as foreign by the mononuclear phagocytic system and are then removed from the bloodstream in extra-vascular sites, such as the spleen and lymph nodes, justifying the occurrence of splenome - galy and lymphadenomegaly in cases of immune-media - ted hemolytic anemia. Trough the examination of blood smears stained for hemoparasite investigation, structures compatible with R. vitalii were observed within leukocytes and extracellularly (Figure 2). Figure 2. Presence of Rangelia vitalii forms within leukocytes (A) and in the extracellular environment (B), observed under light microscopy at 1000x magnifcation. Eritrogram 1Eritrogram 2Eritrogram 3Reference valueRed blood cells 4.5 2.6 1.96 5-10 millions/ul Hemoglobine 8.65.34.28 - 15 g/dL Hematocrit 26.117.913.624 - 45 % MCV 58.167.869.439 - 55 fL MCHC 3329.630.931 - 35 % Blood platelets 253813300 - 800 (10 3 /µL) PPT 6.24.64.2 g/dL6 - 8 g/dL MCV = Mean corpuscular volume; MCHC = Mean Corpuscular Hemoglobin Concentration; TPP = Total plasma protein. Table 1. Hematological tests of the dog diagnosed with rangeliosis. Te erythrogram of animals infected with R. vitalii typically exhibits fndings characteristic of hemolytic anemia. Tis anemia is generally regenerative, ranging from normocytic/normochromic to macrocytic/hypochromic (Rodrigues et al ., 2022). Morphological abnormalities include anisocytosis, polychromasia, spherocytosis, metarubricytosis, and the presence of Howell-Jolly bodies. Additionally, thrombocytopenia and alterations in biochemical
117 Rangelia vitalii in domestic dog in southern Brazil Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 parameters, such as elevated liver enzyme and bilirubin levels, are also commonly observed (França et al ., 2013).Te main clinical signs presented by animals with rangeliosis include fever, apathy, anemia, jaundice, and splenomegaly (Fighera et al ., 2010). Spontaneous bleeding is also commonly reported, which is correlated with intense platelet consumption, as this protozoan has the ability to parasitize and damage the vascular endothelium, potentially afecting the ears and skin (Soares et al ., 2018). Te clinical and hematological fndings of the animal in this case report are consistent with those described in the literature.After the defnitive diagnosis, treatment was initiated with imidocarb dipropionate (5 mg/kg IM) and doxycycline (10 mg/kg SID). Due to the worsening of anemia and clinical signs, supportive care was also provided, including blood transfusion and fuid therapy, as well as the administration of an antiemetic (maropitant citrate), anti-infammatory (meloxicam), analgesic (tramadol), and vitamin supplements. Te patient remained hospitalized for 7 days, however, due to the worsening of her condition, she ended up dying. Te treatment of rangeliosis involves the use of chemotherapeutic agents to eliminate the protozoan from the host. Among the drugs reported for treating canine rangeliosis are diminazene aceturate, imidocarb dipropionate, and doxycycline (França et al ., 2014). Doxycycline is considered inefective when administered alone (Quadros et al ., 2015; Silva et al ., 2019). However, the combination of doxycycline with imidocarb dipropionate has been found efective in the treatment of rangeliosis (França et al ., 2010). Blood transfusion and Wsupportive fuid therapy are necessary in cases of severe anemia (Loretti, 2012). Corticosteroid therapy may also be required to manage immune-mediated hemolytic anemia. Rangeliosis mainly afects dogs from rural areas or those with access to such environments due to the presence of the tick A. aureolatum (Loretti & Barros, 2005; Fighera, 2007), the vector of the disease. Soares et al. (2018) conducted a study to evaluate the vector competence of Rhipicephalus sanguineus (Latreille, 1906) (tropical and temperate lineages), A. aureolatum , Amblyomma ovale (Koch,1844), Amblyomma tigrinum (Koch, 1844), and Amblyomma sculptum (Berlese, 1888) for R. vitalii . Among the six species, only A. aureolatum ticks were competent to transmit R. vitalii to dogs.Tere is a higher incidence of cases in the southern and southeastern regions of Brazil, as reported by several authors in the states of Santa Catarina (Rodrigues et al ., 2022), Rio Grande do Sul (Gottlieb et al ., 2016; Malheiros et al ., 2016; França et al ., 2010), Paraná (Mongruel et al ., 2017), Rio de Janeiro (Lemos et al ., 2012), and Minas Gerais (Moreira et a l., 2013). Cases have also been reported in Argentina (Eiras et al ., 2014) and Uruguay (Soares et al ., 2015).It is also possible that a wild reservoir exists in the biological cycle of R. vitalii , capable of maintaining this protozoan without causing disease (Loretti et al ., 2003). In the state of Rio Grande do Sul, it is believed that this parasite is sustained in the environment by wild animals such as the crab-eating fox ( Cerdocyon thous ) (Linnaeus, 1758), the pampas fox or “zorro de campo” ( Pseudalopex gymnocercus ) (Fischer, 1814), and the raccoon ( Procyon cancrivorus ) (Cuvier, 1798) (Fredo et al., 2015).Additionally, it is important to consider tick control, as transmission occurs through A. aureolatum , highlighting the need for more rigorous environmental and preventive measures, such as the regular use of ectoparasiticides, particularly in dogs from endemic areas. Silva et al. (2019) emphasize that efective prevention of tick infestations can signifcantly reduce the incidence of rangeliosis, especially in rural regions where dogs have greater exposure to the vector.Tis case report also raises questions about the possible underreporting of rangeliosis in urban areas. Although traditionally associated with rural regions, the expansion of urban areas into previously tick-endemic regions may be contributing to an increase in cases in urban environments.Canine rangeliosis is possibly an underdiagnosed disease, as many professionals are unaware of its existence, which may contribute to the underdiagnosis of R. vitalii infections in dogs. Terefore, it is crucial to consider rangeliosis as a diferential diagnosis in cases presenting with hemorrhage, anemia, thrombocytopenia, and splenomegaly, particularly in patients from rural areas, forested regions, or those with access to such environments. Author contributions: CRediT (Contributor Roles Taxonomy)AF = Alexsander Ferraz LDP = Laura Dias Petricione ETB = Eugênia Tavares Barwaldt
118 Neotropical Helminthology, Vol. 19, Nº1, jan - jun 2025 Ferraz et al. RFO = Renata Fontes Ongaratto TCR = Taís Cezimbra Reichow WPO = Wesley Porto de Oliveira RLS = Rodrigo Leite dos Santos CML = Camila Moura de Lima FRPB = Fábio Raphael Pascoti Bruhm LQN = Leandro Quintana Nizoli Conceptualization : AF Data curation : AF, LDP, ETB Formal Analysis : AF, LDP, ETB, CML, TCR, WPO, RLS, CML, FRPB, LQN Funding acquisition : AF, LDP, ETB, CML, TCR, WPO, RLS, CML, FRPB, LQN Investigation : AF, LDP, ETB, CML, TCR, WPO, RLS, CML, FRPB, LQN Methodology : AF, LDP, ETB, CML, TCR, WPO, RLS, CML, FRPB, LQN Project administration : AF Resources : AF, LDP, LQN Software : AF, LDP, ETB, CML, TCR, WPO, RLS, CML, FRPB, LQN Supervision : AF, LQN Validation : AF, LDP, ETB, CML, TCR, WPO, RLS, CML, FRPB, LQN Visualization : AF, LQN Writing – original draft : AF, LDP, ETB, CML, TCR, WPO, RLS, CML, FRPB, LQN Writing – review & editing : AF, LDP, ETB, CML, TCR, WPO, RLS, CML, FRPB, LQN BIBLIOGRAFIC REFERENCES Borrás, P., Salvador, F., Rinaldi, V., Armitano, R., Farber, M., Sanchez, R., Mori, L., Guillemi, E. (2020). Use of molecular tools for the diagnosis of rangeliosis by Rangelia vitalii in Argentina: A case report. Veterinary Parasitology: Regional Studies and Reports , 21 , 100426.Eiras, D.F., Craviotto, M.B., Baneth, G., & Moré, G. (2014). First report of Rangelia vitalii infection (canine rangeliosis) in Argentina. Parasitology International , 63, 729-734 . Fighera, R.A. (2007). Rangeliose. Acta ScientiaeVeterinariae , 35, 261-263.Fighera, R.A., Souza, T.M., Kommers, G.G., Irigoyen, L.F., & Barros, C.S.L. (2010). Patogênese e achados clínicos, hematológicos e anatomopatológicos da infecção por Rangelia vitalii em 35 cães (1985-2009). Pesquisa Veterinária Brasileira , 30, 974-87. França, R.T., Silva, A.S., Costa, M.M., Paim, F.C., Soares, J.F., Labruna, M.B., Mazzanti, C.M., & Lopes, S.T. (2013). Hematologic and bone marrow changes in dogs experimentally infected with Rangelia vitalii. Veterinary Clinical Pathology , 42, 31-39.França, R.T., Silva, A.S., Loretti, A.P., Mazzanti, C.M., & Lopes, S.T.A. (2014). Canine rangeliosis due to Rangelia vitalii : From frst report in Brazil in 1910 to current day – A review. Ticks and Tick-borne Diseases, 5, 466-474.França, R.T., Silva, A.S., Paim, F.C., Costa, M.M., Soares, J.F., Mazzanti, C.M., & Lopes, S.T.A. (2010). Rangelia vitalli in dogs in southern Brazil. Comparative Clinical Pathology , 19, 383-387.Fredo, G., Bianchi, M.V., Andrade, C.P., Souza, S.O., Leite-Filho, R.V., Bandinelli, M.B., Amori, D.B., Driemeier, D., & Sonne, L. (2015). Natural Infectionof Wild Canids ( Cerdocyon thous and Lycalopex gymnocercus ) with the Intra endothelial Piroplasm Rangelia vitalii in Southern Brazil. Journal Wildl Disease , 51, 880-884 . Gottlieb, J., André, M. R., Soares, J. F., Gonçalves, L.R., Oliveira, M., Costa, M. M., Labruna, M.B., Bortolini, C.E., Machado, R.Z., & Vieira, M.I.B. (2016). Rangelia vitalii , Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil. Revista Brasileira de Parasitologia Veterinária , 25, 172–178.
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