Abstract The objective of this investigation was to detect positive cases of 3 species of hemopathogens, from the records of 300 dog samples, without distinction of breed, sex and age from the metropolitan area of Asunción, using the conventional PCR (cPCR) technique. for the detection of Ehrlichia spp. and Anaplasma platys from whole blood and Leishmania spp. in whole blood and spinal aspirate; in addition to blood serum for the detection of antibodies to Leishmania spp. and Ehrlichia spp. using immunochromatographic tests, processed by the Biotechnology Laboratory Division of the Faculty of Veterinary Sciences of the National University of Asunción during the first semester of 2021. Out of the 300 records analyzed, 66 were positive for at least one hemoparasite. The detection of positive cases, according to the type of microorganism identified, was for Leishmania spp. 13%, Ehrlichia spp. 9% and Anaplasma platys 0%. Regarding the type of samples analyzed, using cPCR or immunochromatography interchangeably, positive cases were mostly through whole blood 15%, followed by blood serum 6% and finally bone marrow aspirate 3%. Finally, 27 breeds were recorded, of which the mixed breed had the highest number of positive results for hemoparasites (40/66). The findings from this study provide relevant data for understanding the presence and distribution of hemoparasites in canines in the metropolitan area.

Resumen La presente investigación tuvo como objetivo detectar casos positivos de 3 especies de hemopatógenos, a partir de los registros de 300 muestras de perros, sin distinción de raza, sexo y edad procedentes del área metropolitana de Asunción, empleando la técnica PCR convencional (cPCR), para la detección de Ehrlichia spp. y Anaplasma platys a partir de sangre entera y Leishmania spp. en sangre entera y aspirado medular; además de suero sanguíneo para la detección de anticuerpos para Leishmania spp. y Ehrlichia spp. utilizando test inmunocromatográfico, procesadas por la División Laboratorio de Biotecnología de la Facultad de Ciencias Veterinarias de la Universidad Nacional de Asunción durante el primer semestre del 2021. De las 300 muestras analizadas, 66 fueron positivas para al menos un hemopatógeno. La detección de casos positivos, según tipo de microorganismo identificado fue, para Leishmania spp. del 13 %, Ehrlichia spp. del 9 % y Anaplasma platys del 0 %. En cuanto al tipo de muestras analizada, empleando cPCR o inmunocromatografía indistintamente, los casos positivos fueron mayoritariamente a través de sangre entera 15 %, seguida por suero sanguíneo 6 % y finalmente aspirado medular 3 %. Por último, fueron registradas 27 razas, de las cuales la mestiza presentó mayor número de positivos a hemoparásitos (40/66). Los hallazgos encontrados a partir del presente trabajo aportan datos relevantes para el conocimiento de la presencia de hemoparásitos en caninos del área metropolitana y su distribución.

Abstract The main objective of the research work was the identification of Rickettsias sp. and hemoparasites in ticks of the genus Rhipicephalus sanguineus in dogs. A total of 62 adult ticks carried by 62 dogs from the Concepción neighborhood (urban area) of the city of San Juan Bautista, department of Misiones, Paraguay, in 2017, were analyzed (regardless of age, race and sex). A direct smear of the tick hemolymph extracted from the canines and stained with Wright staining was performed. The results revealed that 13% of the ticks were positive for Rickettsias sp. and hemoparasites. In detail, 6.5% showed the presence of elemental bodies of Ehrlichia spp., while another 6.5% presented vermiform bodies of Babesia spp. The remaining 87% of the samples were negative for Rickettsias sp. and hemoparasites. Regarding the canines whose ticks tested positive for Rickettsias sp. and hemoparasites, 4 canines that represent 50 % were male, while 4 canines corresponded to the female sex that represented 50 %. Regarding the age range of the canines sampled, 25 % represented canines under 1 year, 50 % canines between 1 and 5 years old, and 25 % canines older than 5 years. Regarding the breed of canines whose ticks were positive for Rickettsias sp. and hemoparasites, 37,5 % of the canines were purebred and 62,5 % were mixed breed.

Resumen El objetivo principal de esta investigación fue identificar la presencia de Rickettsias sp. y hemoparásitos en garrapatas de la especie Rhipicephalus sanguineus en perros. Se analizaron un total de 62 garrapatas adultas cargadas recolectadas de 62 perros del barrio Concepción, zona urbana de la ciudad de San Juan Bautista, en el departamento de Misiones, Paraguay, durante el año 2017. La muestra incluyó perros de diversas edades, razas y sexos. Para el análisis, se realizó un frotis directo de la hemolinfa de las garrapatas extraídas de los caninos, y posterior tinción Wright. Los resultados revelaron que el 13% de las garrapatas resultaron positivas para Rickettsias sp. y hemoparásitos. En detalle, el 6,5% mostró la presencia de cuerpos elementales de Ehrlichia spp., mientras que otro 6,5% presentó cuerpos vermiformes de Babesia spp. El 87% restante de las muestras resultaron negativas para Rickettsias sp. y hemoparásitos. En cuanto a los perros cuyas garrapatas dieron positivo para Rickettsias sp. y hemoparásitos, el 50% eran machos, y el otro 50% eran hembras. En cuanto a la edad de los perros muestreados, el 25% tenían menos de 1 año, el 50% tenían entre 1 y 5 años, y el 25% restante tenían más de 5 años. Con relación a la raza de los perros cuyas garrapatas resultaron positivas para Rickettsias sp. y hemoparásitos, el 37,5% eran de raza pura y el 62,5% eran de raza mestiza

Abstract Babesiosis and Anaplasmosis are diseases associated with economic losses; ticks and blood-sucking flies are important zoonotic vectors and reservoirs. This study aimed to investigate the presence of anti-Babesia spp. and anti-Anaplasma marginale antibodies using enzyme-linked immunosorbent assay (ELISA), in ruminants at the Catimbau National Park. Blood samples were collected from 119 sheep, 119 goats, and 47 cattle. Rhipicephalus microplus ticks were collected from cattle. ELISA showed seropositivity of 34% (16/47), 20.3% (24/119), and 16% (19/119) for anti-Babesia bovis; 34% (16/47), 15.2% (18/119), and 9% (7/119) for anti-Babesia bigemina; and 34% (16/47), 35.6% (42/119), and 17% (20/119) for anti-A. marginale antibodies in cattle, goats, and sheep, respectively. The information collected using an epidemiological questionnaire showed that mostly are breed in a semi-intensive system, with access to Caatinga vegetation. The circulation of B. bovis, B. bigemina, and A. marginale was confirmed. Thus, based on the prevalence, this suggests this is an enzootic instability area and is prone to outbreaks. losses bloodsucking blood sucking reservoirs antiBabesia anti Babesia spp antiAnaplasma Anaplasma enzymelinked enzyme linked ELISA, , (ELISA) Park 11 sheep goats 4 cattle 34 16/47, 1647 16/47 16 (16/47) 203 20 3 20.3 24/119, 24119 24/119 24 (24/119) 19/119 19119 19 (19/119 bovis 152 15 2 15.2 18/119, 18119 18/119 18 (18/119) 9 7/119 7119 7 (7/119 bigemina 356 35 6 35.6 42/119, 42119 42/119 42 (42/119) 17 20/119 20119 (20/119 antiA. antiA A anti-A respectively semiintensive semi intensive system vegetation B confirmed Thus prevalence outbreaks (ELISA 1 164 16/4 (16/47 20. 2411 24/11 (24/119 19/11 1911 (19/11 15. 1811 18/11 (18/119 7/11 711 (7/11 35. 4211 42/11 (42/119 20/11 2011 (20/11 16/ (16/4 241 24/1 (24/11 19/1 191 (19/1 181 18/1 (18/11 7/1 71 (7/1 421 42/1 (42/11 20/1 201 (20/1 (16/ 24/ (24/1 19/ (19/ 18/ (18/1 7/ (7/ 42/ (42/1 20/ (20/ (16 (24/ (19 (18/ (7 (42/ (20 (1 (24 (18 ( (42 (2 (4

Resumo A Babesiose e a Anaplasmose são doenças associadas a grandes perdas econômicas, sendo carrapatos e moscas sugadoras de sangue importantes vetores e reservatórios zoonóticos dessas enfermidades. Este estudo teve como objetivo investigar a circulação de anticorpos anti-Babesia spp. e anti-Anaplasma marginale usando Ensaio de Imunoabsorção Enzimática (ELISA), e Babesia spp. em ruminantes no Parque Nacional do Catimbau. Foram coletados ectoparasitos e amostras de sangue de 119 ovinos, 119 caprinos e 47 bovinos. Carrapatos Rhipicephalus microplus foram coletados em bovinos. O teste ELISA mostrou soropositividade de 34% (16/47), 20,3% (24/119) e 16% (19/119) para anti-Babesia bovis; 34% (16/47), 15,2% (18/119) e 9% (7/119) para anti-Babesia bigemina; e 34% (16/47), 35,6% (42/119) e 17% (20/119) para anti-A. marginale em bovinos, caprinos e ovinos, respectivamente. As informações coletadas por meio de questionário epidemiológico mostraram a maioria com sistema semi-intensivo, com acesso à vegetação de Caatinga. A circulação de B. bovis, B. bigemina e A. marginale foi confirmada. Assim, com base na prevalência, essa área foi sugerida como de instabilidade enzoótica e propensa a surtos. econômicas enfermidades antiBabesia anti spp antiAnaplasma Anaplasma ELISA, , (ELISA) Catimbau 11 ovinos 4 bovinos 34 16/47, 1647 16/47 16 (16/47) 203 20 3 20,3 24/119 24119 24 (24/119 19/119 19119 19 (19/119 bovis 152 15 2 15,2 18/119 18119 18 (18/119 9 7/119 7119 7 (7/119 356 35 6 35,6 42/119 42119 42 (42/119 17 20/119 20119 (20/119 antiA. antiA anti-A respectivamente semiintensivo, semiintensivo semi intensivo, intensivo semi-intensivo Caatinga B confirmada Assim prevalência surtos (ELISA 1 164 16/4 (16/47 20, 24/11 2411 (24/11 19/11 1911 (19/11 15, 18/11 1811 (18/11 7/11 711 (7/11 35, 42/11 4211 (42/11 20/11 2011 (20/11 16/ (16/4 24/1 241 (24/1 19/1 191 (19/1 18/1 181 (18/1 7/1 71 (7/1 42/1 421 (42/1 20/1 201 (20/1 (16/ 24/ (24/ 19/ (19/ 18/ (18/ 7/ (7/ 42/ (42/ 20/ (20/ (16 (24 (19 (18 (7 (42 (20 (1 (2 ( (4

Abstract Variations in hematological profile in reptiles can be caused by multiple factors, including parasites presence. Our goals were to identify and morphologically describe blood cells of Liolaemus pacha and analyze their relationship with sex, body condition, individual reproductive/post-reproductive period and mite infestation. Blood smear analyses do not indicate the presence of hemoparasites, suggesting that the mites Neopterygosoma do not serve as vectors for these organisms, as has been proposed for other genera of ectoparasitic mites. In post-reproductive period, there was a reduction in specimens’ body condition and a higher leukocyte count in uninfected lizards. This could be a consequence of the testosterone effects, in higher concentration during the reproductive season, which can increase the metabolic rate, decreasing feeding rate. Infested and non-infested lizards showed no differences in body condition, as well as in leukocyte count, hence the host’s immune system could be developing infestation tolerance. Infested specimens had a higher count of monocytes, thrombocytes, heterophils and lymphocytes. Based on cells function, mites’ effect could be associated with inflammatory processes, allergic reactions or infectious diseases. These results suggested a complex interaction between lizards’ hematological parameters and factors associated to ectoparasites or body conditions. We consider this work as a diagnostic tool for genus Liolaemus, to evaluate health quality, with relevance to the conservation or management of this lizard’s genus. sex reproductive/postreproductive reproductivepostreproductive reproductive/post post hemoparasites organisms postreproductive effects season rate noninfested non infested hosts host s tolerance monocytes thrombocytes lymphocytes function processes diseases conditions quality lizard reproductivepost

Resumen Introducción: Los tripanosomas son hemoparásitos que pueden observarse circulando en la sangre periférica de las aves. Los estudios parasitológicos en aves en el medio natural son escasos, pero son importantes para la investigación relacionada con la transmisión, el mantenimiento del ciclo biológico y la abundancia, entre otros aspectos parasitológicos. Objetivo: Describir infecciones por Trypanosoma sp. en aves de la Amazonia brasileña, así como la prevalencia, características morfológicas y morfométricas de este hemoparásito. Métodos: En la Floresta Nacional de Tapajós, capturamos un total de 125 aves, la mayoría del orden Passeriformes. Obtuvimos muestras de sangre por punción de la vena cubital del ala con agujas estériles de insulina. Con un tubo capilar microhematocrito, obtuvimos alícuotas de sangre. Realizamos frotis de sangre por triplicado y teñimos con el método de Giemsa. Visualizamos los morfotipos de Trypanosoma sp. al microscopio óptico con lentes objetivos de 40 X y 100 X. Para determinar las características morfométricas de Trypanosomatidae, usamos el paquete informático Zen Blue Edition 2. Resultados: Observamos la presencia de hemoparásitos en la forma tripomastigote en ejemplares de la familia de aves Thamnophilidae, Dendrocolaptidae y Conopophagidae, con baja prevalencia. Solo detectamos un morfotipo de Trypanosoma sp. Conclusión: Reportamos la infección por Trypanosoma sp. en especies de importancia ecológica, como Phlegopsis nigromaculata en peligro de extinción en Brasil. La morfología y morfometría del morfotipo encontrado puede contribuir con descripciones más detalladas de estos hemoparásitos.

Abstract Introduction: Trypanosomes are hemoparasites that can be observed circulating in the peripheral blood of birds. Parasitological studies in birds in their natural environment are neglected, but are important for research relating to transmission, maintenance of the biological cycle, and abundance, among other parasitological aspects. Objective: To describe infections by Trypanosoma sp. in birds in the Brazilian Amazon, as well as the prevalence, morphological and morphometric characteristics of this hemoparasite. Methods: In the Tapajós National Forest, we captured a total of 125 birds, mostly from the order Passeriformes. We obtained blood samples from the ulnar vein using sterile insulin needles, and aliquots of blood using a microhematocrit capillary tube. We made blood smears in triplicate and stained with the Giemsa method. We viewd the morphotypes of the Trypanosoma sp. under the light microscope with objective lenses of 40 X and 100 X. To determine the morphometric characteristics of Trypanosomatidae, we used the Zen Blue Edition 2 software package. Results: We observed the presence of hemoparasites in the trypomastigote form in specimens of Thamnophilidae, Dendrocolaptidae and Conopophagidae, with low prevalence. Only one morphotype of Trypanosoma sp. was detected and measurement. Conclusions: We report the infection by Trypanosoma sp. in species of ecological importance, such as Phlegopsis nigromaculata, endangered in Brazil. The morphology and morphometry of the morphotype found could contribute to more detailed descriptions of these hemoparasites.

Abstract: The aim of this study was to evaluate the hematocrit and body temperature values of bovines that tested positive and negative to Babesia spp from two livestock establishments. For this purpose, the blood of 79 bovines with no distinction of sex or breed, older than 2 years of age was evaluated by the blood smear and Giemsa staining; 24 of which resulted positive to Babesia spp and 55 resulted negative. Once the blood samples were obtained, they were sent refrigerated in sterile tubes with anticoagulant EDTA for the hemoparasites observation and hematocrit medition in the Parasitology laboratory of the Faculty of Veterinary Sciences, from the Universidad Nacional de Asuncion. The following results were obtained: the hematocrit means were found within the reference interval in both positive and negative subjects, with no significant differences being found when comparing both groups (p= 0,27), with the hematocrit value resulting higher in positives than in negatives. On the other hand, the temperature means were found above the reference interval in both positive and negative subjects, with significant differences being found when comparing both groups (p= 0,04), with the temperature resulting higher in negatives than in positives.

Resumen: El objetivo del presente trabajo fue evaluar los valores de hematocrito y temperatura corporal en bovinos positivos y negativos a Babesia spp de dos establecimientos ganaderos. Fueron evaluados 79 animales de la especie bovina, sin distinción de sexo y raza, mayores a 2 años de edad; de los cuales 24 presentaron un diagnóstico positivo a Babesia spp por el método de frotis sanguíneo y tinción con Giemsa y 55 presentaron un diagnóstico negativo. Para ello se obtuvieron muestras de sangre, que fueron remitidas refrigeradas en tubos estériles con anticoagulante ácido etilendiaminotetraacético (EDTA) para la observación de los hemoparásitos y médición del valor hematocrito al laboratorio de Parasitología de la Facultad de Ciencias Veterinarias de la Universidad Nacional de Asunción. Obteniéndose los siguientes resultados: las medias del hematocrito se encontraron dentro del intervalo de referencia tanto en positivos como en negativos, no habiendo diferencias significativas al comparar ambos grupos (p= 0,27), resultando el valor hematocrito mayor en positivos que en negativos. Por su parte, las medias de la temperatura se encontraron sobre el intervalo de referencia tanto en positivos como en negativos, habiendo diferencias significativas al comparar ambos grupos (p= 0,04), resultando la temperatura mayor en negativos que en positivos.

Abstract Trachemys dorbigni is the most abundant freshwater turtle species in Rio Grande do Sul, southern Brazil. Chelonians are known to host a wide variety of pathogens, including viruses, bacteria, hemoparasites and helminths. Among these, nine genera of polystomatid flatworms (Monogenoidea; Polystomatidae) infect freshwater turtles: Apaloneotrema, Aussietrema, Fornixtrema, Manotrema, Pleurodirotrema, Polystomoidella, Polystomoides, Uropolystomoides and Uteropolystomoides. However, little is known about the biology of these parasites in the Neotropical Realm. Through investigative cystoscopy, specimens of Polystomatidae were located inside the urinary bladder of the host T. dorbigni. Retrieved specimens were fixed and stained whole mounts prepared for taxonomic identification. In the present paper, a new species of Polystomoides (Monogenoidea: Polystomatidae) parasitizing the urinary bladder of a freshwater turtle of the species T. dorbigni in Brazil is described. Polystomoides santamariensis n. sp. differs from the congeneric species on the length of the genital spines, which are longer. Given the enormous diversity of freshwater turtles around the world, it is likely that a large number of chelonian polystomatids are still unknown. Sul pathogens viruses bacteria helminths Monogenoidea (Monogenoidea Apaloneotrema Aussietrema Fornixtrema Manotrema Pleurodirotrema Polystomoidella Uteropolystomoides However Realm cystoscopy T identification paper described n sp spines longer world unknown

Resumo Trachemys dorbigni é a espécie de tartaruga de água doce mais abundante do Rio Grande do Sul, no Sul do Brasil. Sabe-se que os quelônios são hospedeiros de uma grande variedade de patógenos, incluindo vírus, bactérias, hemoparasitos e helmintos. Dentre estes, oito gêneros de Polystomatidae (Platyhelminthes: Monogenoidea) infectam tartarugas de água doce: Apaloneotrema, Aussietrema, Fornixtrema, Manotrema, Pleurodirotrema, Polystomoidella, Polystomoides, Uropolystomoides e Uteropolystomoides. No entanto, pouco ainda se sabe sobre a biologia destes parasitos na Região Neotropical. Durante uma citoscopia investigativa, foram observados espécimes de Polystomatidae localizados no interior da bexiga urinária de T. dorbigni. Os espécimes coletados foram fixados e corados para identificação taxonômica. No presente artigo, uma nova espécie de Polystomoides (Monogenoidea: Polystomatidae) parasitando a bexiga urinária de tartaruga de água doce da espécie T. dorbigni no Brasil é descrita. Polystomoides santamariensis n. sp. difere das demais espécies no tamanho dos espinhos genitais, que são maiores. Devido a enorme diversidade de tartarugas de água doce ao redor do mundo, é provável que um grande número de polistomatídeos de quelônios ainda seja desconhecido. Sabese Sabe patógenos vírus bactérias helmintos estes Platyhelminthes (Platyhelminthes Monogenoidea Apaloneotrema Aussietrema Fornixtrema Manotrema Pleurodirotrema Polystomoidella Uteropolystomoides entanto Neotropical investigativa T taxonômica artigo (Monogenoidea descrita n sp genitais maiores mundo desconhecido

Abstract Based on the genetic, morphological, and morphometric data of blood gamonts, we identified Hepatozoon parasites in colubrid snakes sampled in the Eastern Amazon region. Hepatozoon trigeminum was detected in the mussurana snake Clelia clelia and exhibited wide and elongated gamonts (mean dimensions: 14.25±0.65 × 4.31±0.43 μm) with an evident parasitophorous vacuole. Hepatozoon odwyerae sp. nov. was described in the indigo snake Drymarchon corais, whose gamonts have elongated and thin bodies (mean dimensions: 13.41±0.79 × 3.72±0.35 μm) with one end more tapered than the other. Phylogenetic analyses, based on the amplification of a 441 bp fragment of the 18S rRNA gene, revealed that the novel sequences of Hepatozoon spp. from our study were closely related to hemogregarine lineages found in lizards and snakes from Brazil, forming a well-supported monophyletic clade with them. The present study provides the first species description of Hepatoozon in D. corais and a new record of a host species for C. clelia using the integrated taxonomic data. We also highlight the importance of further investigations into the diversity of Hepatozoon in snakes, a rich but underestimated group of parasites, especially in the Amazonian biome. genetic morphological region mean dimensions 1425065 14 25 0 65 14.25±0.6 431043 4 31 43 4.31±0.4 μm vacuole sp nov 1341079 13 41 79 13.41±0.7 372035 3 72 35 3.72±0.3 other analyses 44 S gene spp Brazil wellsupported well supported them D C biome 142506 1 2 6 14.25±0. 43104 4.31±0. 134107 7 13.41±0. 37203 3.72±0. 14250 14.25±0 4310 4.31±0 13410 13.41±0 3720 3.72±0 1425 14.25± 431 4.31± 1341 13.41± 372 3.72± 142 14.25 4.31 134 13.41 37 3.72 14.2 4.3 13.4 3.7 14. 4. 13. 3.

Resumen El parasitismo sanguíneo en aves de México es un fenómeno poco estudiado y su presencia en muchas especies de aves es desconocida. En este estudio, se compararon la prevalencia y parasitemia de hemosporidios en la época reproductiva (húmeda) y no reproductiva (seca) de las palomas turcas (Streptopelia decaocto) y huilotas (Zenaida macroura) del norte de México. Se analizó la sangre de 40 aves de cada especie, colectadas entre 2013 y 2014. El diagnóstico de hemoparásitos se realizó mediante técnicas de microscopía y reacción en cadena de la polimerasa (PCR). La prevalencia de hemoparásitos fue de 87.5 % (IC 95%=78.3-93.3). La parasitemia promedio fue de 7.03 (IC 95 %= 5.68-9.04) hemoparásitos por cada 10,000 eritrocitos infectados. La prevalencia y parasitemia fueron mayores para Haemoproteus sp. que en Plasmodium sp y microfilarias. Las tasas de prevalencia no variaron entre especies de aves, ni entre épocas del año. No obstante, la estacionalidad parece ser un factor importante en la parasitemia. La especie que obtuvo mayores tasas de parasitemia fue Z. macoura. Se necesitan más estudios para comprender los mecanismos que asocian la parasitemia de esta especie con respecto a otras especies de columbiformes.

Abstract Blood parasitism in Mexican birds is an impoverished-studied phenomenon, and its presence in many species of birds is unknown. In this study, the prevalence and parasitaemia of hemosporidia were compared in the breeding (wet) and non-breeding (dry) seasons in the Eurasian collared doves (Streptopelia decaocto) and Mourning ones (Zenaida macroura) in northern Mexico. The blood of 40 birds of each species collected between 2013 and 2014, was analyzed. The diagnosis of hemoparasites was made by microscopy and Polymerase Chain Reaction (PCR) techniques. The prevalence of hemoparasites was 87.5% (CI 95% = 78.3-93.3). The mean parasitaemia was 7.03 (CI 95% = 5.68- 9.04) hemoparasites per 10,000 infected erythrocytes. The prevalence and parasitaemia were higher for Haemoproteus sp. than for Plasmodium sp and microfilariae. The prevalence rates did not vary between bird species, nor between times of the year. However, seasonality seems to be an important factor in parasitaemia. The species that obtained the highest rates of parasitaemia was Z. macoura. More studies are needed to understand the mechanisms that associate parasitaemia in this species with respect to other columbiform species.

Abstract Bovine anaplasmosis, babesiosis and trypanosomiasis are diseases of economic importance caused by Anaplasma marginale, Babesia bovis and/or Babesia bigemina, and Trypanosoma vivax, respectively. In this work, protocols received during 2012-2019 in the laboratory of hemoparasites of the EEA Rafaela with a presumptive diagnosis of one of these three diseases were analyzed. The confirmatory diagnosis was made by microscopic observation of the causal agent. Clinical signs and results of serological and molecular tests were used as complementary information. Association between cases of anaplasmosis (dependent variable), the season of the year and area with or without the presence of Rhipicephalus microplus (independent variables) was analyzed with a generalized linear model using a binomial distribution. 50% of the protocols came from Santa Fe. Confirmatory diagnosis was achieved in 48% (213/441) of the protocols, of which 73% were positive for anaplasmosis, 16% for babesiosis and 11% for trypanosomiasis. In the R. microplus free area, 55 cases of anaplasmosis were recorded. The proportion of anaplasmosis cases was significantly higher in autumn compared to summer (p<0.05). No significant differences were observed when anaplasmosis cases, the season of the year and area were analyzed together. The occurrence of anaplasmosis cases in areas considered naturally free of the disease confirms its expansion. Outbreaks of bovine babesiosis occurred in areas with R. microplus. Bovine trypanosomiasis is also an expanding disease and should be considered in the differential diagnosis of hemoparasitic diseases.

Resumen Anaplasmosis, babesiosis y tripanosomiasis de los bovinos son enfermedades de importancia económica provocadas por Anaplasma marginale, Babesia bovis y/o Babesia bigemina, y Trypanosoma vivax, respectivamente. En este trabajo se analizaron los protocolos recibidos durante 2012-2019 en el laboratorio de hemoparásitos de la EEA Rafaela con diagnóstico presuntivo de alguna de estas tres enfermedades. Se realizó diagnóstico etiológico de los casos mediante la observación microscópica del agente causal. Signos clínicos y resultados de pruebas serológicas y moleculares se consideraron como información complementaria. Se analizó la relación entre casos de anaplasmosis (variable dependiente), estación del año y zona con o sin presencia de Rhipicephalus microplus (variables independientes) mediante modelo lineal generalizado utilizando distribución binomial. El 50% de los protocolos provino de la Provincia de Santa Fe. Se arribó a un diagnóstico etiológico en el 48% (213/441) de los protocolos, de los cuales un 73% resultó positivo a anaplasmosis, 16% a babesiosis y 11% a tripanosomiasis. En zona libre de R. microplus se registraron 55 casos de anaplasmosis. La proporción de casos de anaplasmosis resultó significativamente mayor en otoño respecto a verano (p<0,05). El análisis conjunto de casos de anaplasmosis, estación del año y zona, no resultó significativo. La ocurrencia de casos de anaplasmosis en zonas consideradas naturalmente libres de la enfermedad confirma su estatus en expansión. Los brotes de babesiosis bovina continúan circunscriptos a las regiones con R. microplus. La tripanosomiasis bovina también es una enfermedad en expansión y debe ser considerada en el diagnóstico diferencial de enfermedades anemizantes.

RESUMO: O puma (Puma concolor Linnaeus, 1771) tem a maior distribuição entre os felídeos das Américas e é encontrado em todos os biomas do Brasil. No entanto, poucos estudos têm se concentrado nos hemoparasitos nesta espécie. Cytauxzoon felis, um hemoparasito que pode infectar gatos domésticos, também foi descrito em felídeos selvagens no Brasil. A saber, este estudo é o primeiro diagnóstico de infecção natural e detecção molecular de C. felis em um P. concolor do estado de Goiás. Este animal apresentou anemia arregenerativa e inclusão de piroplasmídeos nos glóbulos vermelhos. A amplificação do fragmento de 551 pb da sequência parcial do gene Piroplasmorida 18S rRNA foi 100% idêntica a sequências correspondentes de C. felis disponíveis no GenBank. Nenhum tratamento específico para citauxzoonose foi administrado e, após a reabilitação, o animal foi reintroduzido na natureza. Essa descoberta fornece algumas evidências de que P. concolor pode atuar como um hospedeiro natural do parasito. A epidemiologia, vetor e patogenicidade deste hemoparasito em gatos selvagens e domésticos no Brasil merecem uma investigação mais aprofundada.

ABSTRACT: The puma (Puma concolor Linnaeus, 1771), the most widely distributed felid species in the Americas, can be found in all Brazilian biomes. Nevertheless, few studies have focused on hemoparasites in this species. Cytauxzoon felis, a hemoparasite that can infect domestic cats, has also been described in wild felids in Brazil. To the best of our knowledge, this study is the first to diagnose the natural infection and molecular detection of C. felis in a P. concolor in the state of Goiás. This animal presented non-regenerative anemia and inclusion suggestive of piroplasmids within red blood cells. The amplified 551 bp fragment of partial Piroplasmida 18S rRNA gene sequence was 100% identical to corresponding sequences of C. felis available in GenBank. No specific treatment for cytauxzoonosis was administered, and after rehabilitation, the animal was reintroduced into the wild. This finding provides some evidence that P. concolor may act as a natural host of the parasite. The epidemiology, vector and pathogenicity of this hemoparasite in wild and domestic cats in Brazil deserves further investigation.

Resumen Introducción: En anfibios, la sangre puede actuar como un tejido hematopoyético. Sin embargo, el conocimiento acerca de las características hematológicas es escaso y no hay información que permita un análisis acerca de las posibles explicaciones a este rasgo fisiológico. Objetivo: La intención de este estudio fue evaluar la relación entre la presencia de eritroblastos, mitosis de glóbulos rojos (GRs) y la infección por hemoparásito en sangre periférica de anfibios. Métodos: Se muestrearon 116 anfibios (31 especies) en seis localidades de Colombia. Se tomaron muestras de sangre mediante punción cardiaca o punción a la vena maxilar. Se prepararon extendidos sanguíneos, se fijaron y tiñeron con Giemsa para su posterior análisis por microscopía. Se analizaron variables como porcentaje de GRs inmaduros, células mitóticas en sangre periférica e infección por hemoparásitos. Los datos fueron analizados mediante el test de rango de Willcoxon y el test exacto de Fisher. Resultados: sesenta y dos individuos evidenciaron mitosis en sangre periférica y dichas mitosis compartían características morfológicas con GRs inmaduros. La prevalencia general de parásitos fue del 30.1 %, distribuido de la siguiente forma: Trypanosoma (24.1 %), Hepatozoon-like (6 %), Dactylosoma (4.3 %), Karyolysus-like (0.9 %), y Filarioidea (2. 6 %). Hay una asociación positiva entre el porcentaje de GRs inmaduros y la presencia de células mitóticas, también se encontró una relación entre la infección por hemoparásitos y el porcentaje de GRs inmaduros. Conclusiones: En este estudio encontramos que la presencia de parásitos sanguíneos, GRs inmaduros y mitosis de GRs son eventos frecuentes en sangre periférica de anfibios, y nuestros resultados sugieren una asociación entre dichas características. Por tanto, la liberación de GRs inmaduros y la mitosis de estas células en sangre periférica podría ser una respuesta fisiológica a infecciones parasitarias. Posteriores estudios que caractericen la hematología en anfibios y en vida silvestre en general, son deseables.

Abstract Introduction: In amphibians, blood may act as a hematopoietic tissue. However, the knowledge concerning hematological features is scarce, there is not much information that allows an analysis about the possible explanations of this physiological feature. Objective: This study aimed to evaluate the relationship between immature red blood cells (RBCs) mitosis and the presence of blood parasites in amphibians. Methods: We sampled 116 amphibians (31 species) in six Colombian localities. Blood was taken by cardiac puncture or maxillary vein puncture. Smears were prepared, fixed, and Giemsa stained for microscopical analysis. The variables analyzed were the percentage of immature RBCs, mitotic cells in peripheral blood, and blood parasite infection. Data were analyzed using Wilcoxon’s rank test and exact Fisher statistical tests. Results: Sixty-two individuals showed mitosis in peripheral blood, and these mitotic RBCs shared morphological features with immature RBCs. Overall, parasite prevalence was 30.1 %, distributed as follows: Trypanosoma (24.1 %), Hepatozoon-like (6 %), Dactylosoma (4.3 %), Karyolysus-like (0.9 %), and Filarioidea (2.6 %). A positive association between the percentage of immature RBCs and the presence of mitotic RBCs was found, and also between the blood parasite infection and the percentage of immature RBCs. Conclusions: In this study, we found that the presence of blood parasites, immature RBCs, and RBCs mitosis are frequent events in amphibians’ peripheral blood, and our analysis suggests an association between those features. Thus, the release of immature RBCs and the mitosis of those cells in peripheral blood may be a physiological response to blood parasite infection. Further studies characterizing hematology in amphibians and wildlife, in general, are desirable.

Abstract Rhinella major is one of the component species of the Rhinella granulosa group and is distributed in the neotropical region of the South American continent, being found in anthropized environments. The occurrence of trypanosomes in anurans involves a yet unknown diversity of species and hosts. In the present study, we aimed to describe the first record of Trypanosoma sp. at R. major as a new host. Of the species captured, four were positive for trypomastigote forms of Trypanosoma sp. The morphometry of the trypomastigote forms revealed parasitemia by only one morphotype. There is a need for reports and studies of parasite-host relationships in anurans and a lack of records regarding hemoparasite diversity linked to the Rhinella granulosa group.

Abstract Biological interactions play an important role in regulating and maintaining natural populations. Like most interactions, parasitism may be influenced by environmental conditions. Therefore, changes caused by human activity may drastically affect the equilibrium of the assemblages of parasitized organisms (hosts). Herein, we described the composition of hemoparasites of anurans from two distinct environments: forest and oil palm plantations. We identified the most frequent groups of blood parasites, and whether infections differ between habitats (forest and plantation) and between microhabitats (arboreal or terrestrial). We collected 128 anurans, of which 46 (36%) were parasitized by hemoparasites. The genus Trypanosoma spp. was found in 30% (n = 37/128) of the infected anurans in both habitats, recorded mostly in terrestrial anurans in oil palm plantations. Apicomplexa hemoprotozoans were also found in 13% (n=17/128) of the anurans, which mainly were terrestrial species collected in oil palm plantations. There was no difference in parasitism between the two assemblies and between the studied microhabitats. This is the first study that has analyzed the ecological relationship between anurans as hosts and their blood parasites, in a region under intense anthropic pressure, in the Brazilian Amazon.

Resumo Para uma melhor compreensão da epidemiologia e diversidade genética das infecções por hemoprotozoários em mamíferos selvagens, este estudo teve como objetivo a detecção post mortem de DNA de espécies da ordem Piroplasmida (Babesia sp., Cytauxzoon sp. e Theileria sp.) e subordem Adelorina (Hepatozoon sp.), utilizando-se a reação em cadeia pela polimerase, baseada no gene 18S rRNA, seguido de sequenciamento genético de amostras de sangue e baço, coletadas de 164 carcaças de mamíferos selvagens de vida livre e cativos do estado de Mato Grosso. Entre eles, um Leopardus pardalis, três Panthera onca, dois Puma concolor foram positivos para Cytauxzoon sp., e seis Tapirus terrestris testaram positivos para Piroplasmida, enquanto um L. pardalis foi positivo para Hepatozoon sp. Além disso, foi descrito em antas, um piroplasmídeo não caracterizado geneticamente, relacionado à Theileria sp., previamente detectado em gatos do Brasil. Apesar da controvérsia quanto à ameaça epidemiológica desses protozoários, a detecção desses agentes em mamíferos silvestres e cativos, mesmo quando assintomáticos, demonstra a importância do monitoramento, principalmente em hotspots, como no estado de Mato Grosso, para verificar a circulação e a diversidade genética, a fim de antecipar o possível surgimento de doenças e, até mesmo, suas consequências para outros animais, bem como os humanos.

Abstract To a better insight into the epidemiology and genetic diversity of protozoan hemoparasites infections in wild mammals, this study aimed to the post mortem detection of DNA from species of the order Piroplasmida (Babesia sp., Cytauxzoon sp., and Theileria sp.) and suborder Adelorina (Hepatozoon sp.) using polymerase chain reaction based on the 18S rRNA gene followed by genetic sequencing of blood and spleen samples collected from carcasses of 164 free-ranging and captive wild mammals from Mato Grosso state. Among them, one Leopardus pardalis, three Panthera onca, two Puma concolor were positive for Cytauxzoon sp., and six Tapirus terrestris tested positive for Piroplasmida, while one L. pardalis was positive for Hepatozoon sp. Furthermore, an uncharacterized piroplasmid genetically related to Theileria sp. previously detected in cats from Brazil was described in lowland tapirs. Despite the controversy regarding the epidemiological threat of these protozoa, the detection of these tick-borne agents in wild free-living and captive mammals, even when asymptomatic, demonstrates the importance of monitoring, particularly in hotspots such as the state of Mato Grosso, to verify the circulation and genetic diversity, to anticipate the possible emergence of diseases, and even their consequences to other animals as well as humans.