Molecular detection of Anaplasma, Babesia, Theileria, and Trypanosoma infection in cattle and buffaloes in India.

Tick-borne pathogens pose a significant global threat, causing substantial economic losses to the dairy industry. In India, tropical theileriosis, anaplasmosis, babesiosis, and trypanosomiasis are major hemo-parasitic diseases affecting bovines. A cross-sectional study was conducted to determine the prevalence of hemo-parasites in different farms in India. PCR assays were employed to detect carrier status, using gene targets msp1b, tams1, rap-1, ama1, and ITS1 for A. marginale, T. annulata, B. bovis, B. bigemina, and Trypanosoma species, respectively. Out of the 578 apparently healthy animals screened, 30.45% (95% CI: 26.84-34.32%) were infected with at least one hemo-parasite. Cattle showed an overall positivity of 32.87%, while buffaloes had a prevalence of 15.19%, which was statistically significant (p < 0.001). Interestingly, prevalence was higher in indigenous cattle (47.81%) compared to cross-breeds (25.53%) and exotics (14.62%), with a statistically significant difference (p < 0.001). The prevalence of hemo-parasites varied widely among the farms, ranging from 5.77 to 100%. A. marginale was the most prevalent parasite (23.70% of animals), followed by T. annulata (13.67%), Babesia species (1.90%), and Trypanosoma species (1.56%). Enzootic instability was observed in six of the eight farms, indicating a potential for future outbreaks. Co-infection was detected in 60 out of 176 animals positive for hemo-parasites, with 59 animals co-infected with A. marginale and T. annulata, and only one cross-breed cattle infected with both Anaplasma marginale and Babesia bigemina. The findings highlight the prevalence of hemo-parasites in farms, underscoring the need for whole-herd screening, treatment of infected animals, and improvement in farm management practices to prevent production losses caused by these pathogens. Supplementary Information: The online version contains supplementary material available at 10.1007/s12639-024-01673-3.

Molecular detection of Hepatozoon species (Apicomplexa: Hepatozoidae) infecting snakes in the Northeastern region of Argentina.

The genus Hepatozoon Miller (1908) contains a wide range of obligate parasitic organisms with complex life cycles involving vertebrates and hematophagous invertebrates. Despite over 300 species being described, only a small percentage has been characterized in snakes using morphological and molecular techniques. The prevalence of these parasites in snakes is significant, highlighting the need for molecular descriptions in such elusive hosts. Thus, the objective of this study was to determine molecularly the presence of Hepatozoon species in snakes from the Northeastern region of Argentina. Thirty-two specimens of eight snake species (Bothrops alternatus, Dryophylax hypoconia, Erythrolamprus jaegeri coralliventris, Erythrolamprus poecilogyrus, Erythrolamprus semiaureus, Philodryas olfersii latirostris, Pseudablabes (ex Philodryas) patagoniensis and Palusophis (ex Mastigodryas) bifossatus were collected and examined. PCR analysis of the 18S rRNA locus detected four samples (12% prevalence) positive for the presence of Hepatozoon DNA. Phylogenetic analysis positioned the 18S rRNA Hepatozoon sequences obtained in three different clades, one with Hepatozoon musa, another with sequences of Hepatozoon cuestensis, while the third was placed as a sister taxon to a clade including Hepatozoon cevapii and Hepatozoon massardi. This study presents the first documentation of Hepatozoon infecting snakes in Argentina, thereby expanding their distribution within southern South America. Additionally, B. alternatus and Pa. bifossatus are reported as new hosts of Hepatozoon.

Hemopathogens in naturally infected bovine fetuses in Brazil.

The transplacental transmission of parasites and hemoparasites is crucial for understanding the epidemiology of diseases. This study aimed to assess the prevalence of hemopathogens in bovine fetuses at various gestational periods. Samples were obtained from a slaughterhouse in the state of Minas Gerais, Brazil, and a total of 236 fetuses were collected. DNA extracted from blood samples (145) and organ samples (a pool of brain and spleen) (236) underwent a nested PCR (nPCR) assay to detect Babesia spp., Theileria spp., Trypanosoma vivax, Anaplasma marginale, Anaplasma bovis, Anaplasma phagocytophilum, Ehrlichia minasensis, and hemotropic Mycoplasma spp. Additionally, serological analysis of 145 plasma samples was conducted using the indirect fluorescent antibody test-IFAT to detect IgG against Babesia bovis, Babesia bigemina, A. marginale, and Trypanosoma vivax. The observed prevalence of transplacental transmission was 19.3 %, 6.2 %, 42.7 % and 2.7 %, for A. marginale, B. bigemina, 'Candidatus M. haemobos', and Mycoplasma wenyonii, respectively. The prevalence of A. marginale by gestational trimester was 16 % (13/81) in the second trimester and 23 % (14/60) in the third trimester, with no positive samples in the first trimester. Regarding the species B. bovis and B. bigemina, all evaluated animals tested negative by nPCR, and no serological evidence for B. bovis was found by the IFAT. Babesia bigemina demonstrated an overall seroprevalence of 6.2 % (9/145), with 4.8 % (7/145) in the last trimester and 1.3 % (2/145) in the second trimester of pregnancy. In total, 42.7 % (62/145) of blood samples were positive for 'Candidatus M. haemobos', with 42 % (34/81) in the middle trimester, and 43 % (26/60) in the final trimester of pregnancy. Mycoplasma wenyonni was detected in 2.7 % (4/145) blood samples, all in coinfection with 'C. M. haemobos'. The prevalence by pregnancy trimester was 25 % (1/4) in the first trimester; 1.2 % (1/81) in the second trimester and 3.3 % (2/60) in the third trimester of pregnancy. Hemopathogen DNA was detected in fetus blood samples but not the brain or spleen samples. All the samples were negative for T. vivax, Theileria spp., Anaplasma spp. and Ehrlichia spp. Overall, in this study, approximately 70 % of fetuses were positive for one or more of the studied parasites. No significant associations were observed between pairs of pathogens, except 'C. M. haemobos' and A. marginale.

Molecular survey of Piroplasmida, Hepatozoon spp. and Anaplasmataceae in anemic and thrombocytopenic dogs from Uruguay.

Canine tick-borne diseases, such as babesiosis, rangeliosis, hepatozoonosis, anaplasmosis and ehrlichiosis, are of veterinarian relevance, causing mild or severe clinical cases that can lead to the death of the dog. The aim of this study was detecting tick-borne protozoan and rickettsial infections in dogs with anemia and/or thrombocytopenia in Uruguay. A total of 803 domestic dogs were evaluated, and 10% were found positive (detected by PCR) at least for one hemoparasite. Sequence analysis confirmed the presence of four hemoprotozoan species: Rangelia vitalii, Babesia vogeli, Hepatozoon canis and Hepatozoon americanum, and the rickettsial Anaplasma platys. The most detected hemoparasite was R. vitalii, followed by H. canis and A. platys. This is the first report of B. vogeli in Uruguay and the second report of H. americanum in dogs from South America. The results highlight the importance for veterinarians to include hemoparasitic diseases in their differential diagnosis of agents causing anemia and thrombocytopenia.

Unraveling the diversity of Trypanosoma species from Central Mexico: Molecular confirmation on the presence of Trypanosoma dionisii and novel Neobat linages.

Bats are one of the groups of mammals with the highest number of associated Trypanosoma taxa. There are 50 Trypanosoma species and genotypes infecting more than 75 species of bats across five continents. However, in Mexico, the inventory of species of the genus Trypanosoma associated with bats is limited to only two species (Trypanosoma vespertilionis and Trypanosoma cruzi) even though 140 species of bats inhabit this country. Specifically, 91 bat species have been recorded in the state of Veracruz, but records of trypanosomatids associated with this mammalian group are absent. Due to the complex Trypanosoma-bat relationship, the high diversity of bat species in Veracruz, as well as the lack of records of trypanosomatids associated with bats for this state, the aim of this work was to analyze the diversity of species of the genus Trypanosoma and their presence from a bat community in the central area of the state of Veracruz, Mexico. During the period of January to August 2022 in the Tequecholapa Environmental Management Unit where bats were collected using mist nets and blood samples were obtained from their thumbs. We extracted genetic material and amplified a fragment of 800 bp of the 18S ribosomal gene of the genus Trypanosoma by conventional PCR. The positive amplicons were sequenced, and phylogenetic reconstruction was performed to identify the parasite species. A total of 285 bats (149♀, 136♂) belonging to 13 species from 10 genera and a single family (Phyllostomidae) were collected. Twenty-three specimens from six species tested positive for the presence of Trypanosoma dionisii, Trypanosoma sp. Neobat 4, and a potential novelty species provisionally named as Trypanosoma sp. Neobat 6. The results of the present work increase the number of species of the genus Trypanosoma infecting bats in Mexico and in the Neotropical region.

Blood parasites (Kinetoplastida: Trypanosomatidae) infecting birds in the Brazilian Amazon / Parásitos sanguíneos (Kinetoplastida: Trypanosomatidae) que infectan aves en la Amazonía brasileña

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.

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.

Hemoparasites Do Not Affect Life-History Traits and Cellular Immune Response in Treefrog Hosts Boana cordobae.

We provide the first evidence for hemoparasites in the endemic Cordoba treefrog Boana cordobae. We collected 37 adult frogs at 1200 m a.s.l. in the Comechingones Mountains in the Córdoba province (Argentina). Each individual was sexed, then snout-vent length and body mass were recorded, a toe was collected for skeletochronological age determination, and a slide with a blood smear was prepared for hemoparasite screening, before releasing the frogs in situ. A total of 81% (n = 30) of the frogs were infected by hemogregarines and trypanosomes with a high intensity of infections. Dactylosoma was found for the first time in Argentina. Hemoparasites had no significant effect on the leukocyte profile, which we assessed from the May-Grünwald-Giemsa-stained blood smears. The neutrophils/lymphocytes ratio, indicative of stress, was insignificantly higher (0.06) in parasitized frogs than in parasite-free individuals (0.04). Infected frogs were larger than the controls, but this effect vanished when correcting size data for age. Young frogs (first-breeders) dominated the age distribution of parasite-free individuals, suggesting that infection of frogs takes usually place after sexual maturation. Vectors transmitting hemoparasites to B. cordobae remain to be identified. We demonstrate that moderate to high intensities of hemoparasites do not significantly affect the cellular immune response of B. cordobae, or any of the life-history traits studied, nor did they show any external sign of disease.

Molecular Survey of Hepatozoon canis Infection in Domestic Dogs from Sardinia, Italy.

Tick-borne protozoans of the genus Hepatozoon have been associated with infections of domestic and wild animals over the world. The occurrence of these apicomplexan agents in Sardinia has been poorly explored so far. In this study, the occurrence of Hepatozoon spp. has been investigated in domestic dogs from nine cities of Sardinia, Italy. Blood samples from each dog were collected and tested molecularly for the presence of Hepatozoon and Babesia/Theileria DNAs. Out of fifty-one dogs, nine were positive for Hepatozoon species based on the molecular detection of the parasite in blood samples. The phylogenetic relationships of strains detected here were also established. The PCR for amplification of the 18S rRNA fragment gene of Babesia/Theileria spp. did not give amplicons in any of the analyzed samples. Our results report the first molecular confirmation of Hepatozoon canis in Sardinian pet dogs and contribute to better understand the presence of these protozoans on the island. This study highlights the importance of recognizing and predicting the risk levels for the canine population, thus increasing the development of specific control measures. Also, since the distribution of hepatozoonosis is closely related to that of the definitive tick host, Rhipicephalus sanguineus, more accurate studies on Rhipicephalus ticks will be needed due to increasing the epidemiological knowledge of Hepatozoon species on the island.

Health evaluation of street dogs in Guarapuava, Brazil.

Free-roaming dogs are removed from the street by the Municipal Animal Control division/shelter and NGOs to safeguard public health as well as to prevent any suffering dogs may face and to find them homes. However, adoption rates are low and there is a lack of public programs aimed at humane control causing increased numbers of street dogs and overcrowding in shelters, compromising health and welfare. We aimed to evaluate the health of dogs from municipal shelters and independent shelters, and community dogs in the city of Guarapuava through clinical examination and complete blood cell count tests. The main changes found in the clinical examination of 297 dogs were hyperthermia in 4.0%, tachypnea in 20.5%, ocular and oral mucosa hypocorada in 6.4%, and ectoparasite infestation in 42.7%. Changes found in the erythrogram were anemia in 15.1% and polycythemia in 3.4%. In the platelet count, thrombocytopenia was found in 10.1% and thrombocytosis in 10.4%. On the leukogram, leukocytosis was observed in 14.8% and leukopenia in 3%. The results reveal the compromised health of some animals, requiring better diagnostic investigation, since certain clinical and complete blood cell count alterations may be caused by zoonotic agents that can endanger human and animal health.

Molecular detection of hemotropic Mycoplasma and Bartonella species in lice from sheep and goats of Mexico.

The knowledge of lice associated with small ruminants, especially sheep and goats, is scarce. In Mexico, there are historical reports of six species of chewing and sucking lice associated with Capra hircus and Ovis canadensis. However, the reports did not analyze the ecology of the infestations or the presence of potentially pathogenic bacteria. For this reason, the objectives of this study were i) to identify the species of lice associated with sheep and goats in three states of the Mexican Republic, ii) to characterize the infestations, and iii) to identify the presence of bacterial pathogens. From October 2019 to August 2021, six ranches with sheep and goats were sampled in the states of Hidalgo and Veracruz. Hosts were visually inspected, and lice were retrieved with forceps. The specimens were sexed and identified using morphological taxonomic keys. DNA extraction was performed individually, and a fragment of the cytochrome oxidase subunit 1 gene (COI) was amplified for the molecular identification of the specimens. Subsequently, Anaplasma, Bartonella, Ehrlichia, Mycoplasma, and Rickettsia were molecularly detected. Additionally, the infestations were characterized by calculating the prevalence and mean abundances. We collected 563 specimens of three species, Bovicola caprae, Bovicola ovis, and Linognathus africanus. The highest infestation levels were recorded for B. ovis (66.7%; 4.4) from Veracruz. Additionally, two Bartonella species were detected: Bartonella mellophagi in B. ovis and Bartonella capreoli in L. africanus. In contrast, Mycoplasma ovis was detected exclusively in one pool of B. ovis. This study provides new bacterial-ectoparasite associations and highlights the possible role of these neglected ectoparasites as vectors in the populations of sheep and goats from Mexico.

Clinical and epidemiological aspects of the infection by Babesia, Theileria and Trypanosoma species in horses from northeastern Colombia.

Piroplasmosis and trypanosomiasis are debilitating diseases of great economic impact on the equine industry of Latin America. Considering the lack of studies in the northeastern part of Colombia, this study aimed to determine the epidemiological, clinical and genetic features associated with infection of the Babesia, Theileria, and Trypanosoma species in horses from this geographical area. Two hundred and eighty horses from the Arauca, Meta, and Santander departments were molecularly analyzed for infection with Babesia caballi, Theileria equi, Trypanosoma evansi, and Trypanosoma vivax. Furthermore, clinical, epidemiological and entomological analyses were performed on the data sets. Molecular analysis showed 25.7% and 3.9% prevalence for T. equi and T. evansi, respectively, without positive animals for B. caballi and T. vivax. There were no differences in the prevalence of T. equi between departments, whereas T. evansi was detected exclusively in Santander. A total of 633 ticks were collected from 72 horses across the three departments, with 84.7% corresponding to Dermacentor nitens, 10.9% to Amblyomma cajennense (sensu lato) (s.l). and 4.4% to Rhipicephalus microplus. For T. equi, genetic analyses showed that Colombian isolates belong to genotype C of species, along with sequences of Brazil and Mexico. Epidemiological analysis revealed a significant association between tick infestation and lack of vector control with molecular infection of T. equi, whereas clinical analysis revealed a significant reduction in packed cell volume, red blood cells, and mean corpuscular volume in positive animals to this pathogen. Furthermore, molecular infection by T. evansi was associated with epidemiological characteristics in the Santander department. In conclusion, our analysis revealed a moderate infection rate by T. equi of genotype C in horses from northeastern Colombia, which affects their clinical conditions. Control of ticks and treatment of symptomatic animals should be considered to reduce the economic impact associated with these infections in the equine industry.

Molecular characterization of Schellackia parasites in an urban population of sand lizards (Lacerta agilis) from Berlin, Germany.

Lizards are hosts of several taxa of unicellular parasites of the phylum Apicomplexa, including Karyolysus, Schellackia, Lankesterella, and Hepatozoon. Parasite prevalence and the impact of infections on lizard biology remain largely unexplored. In this study, blood parasite infections were investigated in sand lizards (Lacerta agilis) from Berlin, Germany. Eighty-three individuals were investigated, and the detected blood parasites were identified as Schellackia sp. The combination of microscopic and molecular screening revealed a prevalence of 14.5%. Parasitemia values were low and most infections were subpatent. Phylogenetic analysis recovered a close relationship of the Schellackia parasites of this study with Schellackia sp. parasites of different Lacerta and Podarcis lizard species from Spain. Monitoring of Schellackia parasite infections in free-ranging lizards contributes to a better understanding of the distribution, diversity, and phylogenetic relationships of the neglected parasite taxon.

Hematologic variables of free-living Leptodactylus luctator with and without hemoparasites and thrombidiform mites in southern Brazil.

It has been suggested that anuran amphibian parasites can cause clinical signs in situations of environmental imbalance. In the family Leptodactylidae, information about hematology is scarce, although these are well-known tools for the diagnosis and prognosis in clinical practice and potential bioindicators of environmental stress. The objective of this study is to describe Leptodactylus luctator hematology, to report the occurrence of hemoparasites and thrombidiform mites, and to compare the hematological variables under the presence and absence of these organisms. Ectoparasites and heparinized blood samples from 40 free-living specimens of L. luctator were collected for analysis. Hematologic variables and total plasma protein were compared between groups with and without hemoparasites and intradermal mites. As results, structures compatible with hemogregarines, Lankesterella sp., five morphotypes of Trypanosoma spp., microfilaria, Aegyptianella sp., an unidentified intraleukocytic hemoparasite, and frog erythrocytic virus (FEV) inclusion bodies were identified in the blood samples, besides Hannemania spp. intradermal mites. The hemoparasite occurrence was higher than previously reported in other anuran families and locations. Also, L. luctator has smaller red blood cells (RBCs) and white blood cells (WBCs), and a hyposegmentation of the neutrophil nucleus, when compared to many other amphibians. White blood cell, neutrophil, and monocyte counts were higher in animals parasitized by mites. There was no correlation between the number of parasitized RBCs and hematologic variables. This study provides anuran hematologic information, in addition to indicating a host reaction to infestation by Hannemania spp. mites, besides constituting the first record of the distribution of hemoparasites and intradermal mites in L. luctator of the study region.

Detecção molecular de Anaplasma platys, Ehrlichia canise Babesia spp. em cães atendidos em clínica da região metropolitana do estado do Rio de Janeiro / Molecular detection of Anaplasma platys, Ehrlichia canis and Babesia spp. in dogs served at a clinic in the metropolitan region of the state of Rio de Janeiro

O presente estudo teve como objetivo detectar por meio da Reação em cadeia da Polimerase (PCR) a frequência de Ehrlichiacanis, Babesia spp. e Anaplasma platys em cães, relacionando a prevalência dos achados hematológicos aos resultados positivos pela PCR. Foram avaliadas 209 amostras de sangue de cães atendidos em clínica veterinária particular do município de Queimados, RJ, Brasil, no período de julho a outubro de 2014. Foram realizados hemograma completo e extração de DNA para técnica de PCR. Do total de 209 animais, 19,1% (40/209) animais apresentaram resultado positivo para hemoparasitos pela técnica de PCR. Destes, 52,5% (21/40) foram positivos para E. canis, 27,5% (11/40) positivos para Babesia spp. e 10% (4/40) positivos para A. platys. Quatro animais (1,91%), dos 209 testados, foram positivos para pelo menos dois agentes, caracterizando assim coinfecção. Dos 40 cães positivos para algum dos agentes testados, 25 (62,5%) estavam trombocitopênicos. Ou seja, 15 cães (37,5%) reagiram positivos para hemoparasitos, mas não apresentavam trombocitopenia. A anemia foi um achado comum, sobretudo nas infecções por Babesia spp., 100% (11/11) e E.canis, 90,5% (19/21). A técnica de PCR foi um importante método diferencial na detecção das principais hemoparasitoses caninas, juntamente com os achados clínicos e hematológicos para o diagnóstico preciso da infecção em questão.

The present study aimed to detect, by means of Polimerase chain reaction (PCR), the frequency of Ehrlichia canis, Babesia spp. and Anaplasma platys in dogs, relating the prevalence of hematological findings to positive PCR results. A total of 209 blood samples from dogs treated at a private veterinary clinic in the city of Queimados, RJ, Brazil, from July to October 2014 were evaluated. Complete blood count and DNA extraction were performed for the PCR technique. Of the total of 209 animals, 19.1% (40/209) animals were positive for hemoparasites by the PCR technique. Of these, 52.5% (21/40) were positive for E. canis, 27.5% (11/40) were positive for Babesia spp. and 10% (4/40) positive for A. platys. Four animals (1.91%) of the 209 tested were positive for at least two agents, thus characterizing coinfection. Of the 40 dogs positive for any of the agents tested, 25 (62.5%) were thrombocytopenic. That is, 15 dogs (37.5%) were positive for hemoparasites, but did not have thrombocytopenia. Anemia was a common finding, especially in infections by Babesia spp., 100% (11/11) and E. canis, 90.5% (19/21). The PCR technique was an important differential method in the detection of the main canine hemoparasitoses, together with the clinical and hematological findings for the accurate diagnosis of the infection in question.

Filarial infections in lemurs: Evidence for a wide geographical distribution and low host specificity among lemur species.

The relevance of emerging infectious diseases continues to grow worldwide as human activities increasingly extend into formerly remote natural areas. This is particularly noticeable on the island of Madagascar. As closest relatives to humans on the island, lemurs are of particular relevance as a potential origin of zoonotic pathogen spillover. Knowledge of pathogens circulating in lemur populations is, however, very poor. Particularly little is known about lemur hemoparasites. To infer host range, ecological and geographic spread of the recently described hemoparasitic nematode Lemurfilaria lemuris in northwestern Madagascar, a total of 942 individuals of two mouse lemur species (Microcebus murinus [n = 207] and Microcebus ravelobensis [n = 433]) and two rodent species (the endemic Eliurus myoxinus [n = 118] and the invasive Rattus rattus [n = 184]) were captured in two fragmented forest landscapes (Ankarafantsika National Park and Mariarano Classified Forest) in northwestern Madagascar for blood sample examination. No protozoan hemoparasites were detected by microscopic blood smear screening. Microfilaria were present in 1.0% (2/207) of M. murinus and 2.1% (9/433) of M. ravelobensis blood samples but not in rodent samples. Internal transcribed spacer 1 (ITS-1) sequences were identical to an unnamed Onchocercidae species previously described to infect a larger lemur species, Propithecus verreauxi, about 650 km further south. In contrast to expectations, L. lemuris was not detected. The finding of a pathogen in a distantly related host species, at a considerable geographic distance from the location of its original detection, instead of a microfilaria species previously described for one of the studied host species in the same region, illustrates our low level of knowledge of lemur hemoparasites, their host ranges, distribution, modes of transmission, and their zoonotic potential. Our findings shall stimulate new research that will be of relevance for both conservation medicine and human epidemiology.

Prevalence and risk factors associated with Babesia bovis infection in Crioula Lageana cattle.

INTRODUCTION: Bovine babesiosis caused by the protozoan Babesia bovis is a worldwide disease and causes great economic damage to livestock. There are no studies on the epidemiology of this disease in native breeds such as Crioula Lageana cattle raised in the South of Brazil. METHODOLOGY: DNA samples from 311 animals were amplified by polymerase chain reaction (PCR) for the identification of the gene rap-1 (Rhoptry Associated Protein 1) from B. bovis. An epidemiological questionnaire was used to determine the risk factors associated with infection. RESULTS: The prevalence of B. bovis infection was 72% (224/311). Age and tick infestation affected infection. The factors associated with infection were the breeding objective (p = 0.042; CI = 0.746-0.995; OR = 0.861), contact of cattle with other animal species (p = 0.002; CI = 0.517-0.860; OR = 0.484), absence of tick control (p = < 0.001; CI = 0.074-0.480; OR = 0.188) and timing of tick treatment (p = 0.026; CI = 0.673-0.975; OR = 0.810), and these were considered to be factors that can protect against the disease. CONCLUSIONS: The Crioula Lageana cattle breed has near enzootic stability with regards to B. bovis infection.

First report of canine rangeliosis in domestic dogs from different regions of Santa Catarina State, Brazil.

Rangeliosis is the disease caused by Rangelia vitalii, a parasite reported in dogs from southeastern and southern Brazil, Uruguay, Paraguay, and Argentina. This protozoan is transmitted by the ixodid Amblyomma aureolatum, and infects erythrocytes, leukocytes, and vascular endothelial cells of the host. The common clinical signs, such as prostration, fever, anemia, thrombocytopenia, anorexia, weight loss, and dehydration, are also found in other infections, like canine babesiosis and ehrlichiosis. The similar clinical presentation with other diseases, as well as the indistinguishable morphology with intraerythrocytic Babesia canis, complicates the disease diagnostic. In the present study, blood samples from dogs presenting clinical signs compatible with hemoparasitosis were investigated for rangeliosis. The dogs were treated at veterinary clinics in the cities of Blumenau and Lages, in the State of Santa Catarina, Brazil. Blood samples from 17 dogs were analyzed by PCR. The samples were screened by a conventional piroplasma-PCR and the positives confirmed by a specific R. vitalii-qPCR. Two animals (2/17; 11.8%) were positive for R. vitalii, one from Blumenau and the other from Lages. Both animals presented unspecific signs of hemoparasitosis, such as apathy, anemia, and anorexia. The results indicate the necessity of molecular assays for the proper identification of the hemoparasite, and to investigate the real prevalence of rangeliosis in the State of Santa Catarina.

Incidence of hemoparasitic infections in cattle from central and northern Thailand.

Background: Hemoparasites, such as Babesia spp., Theileria spp. and Anaplasma spp., can negatively affect the health of farm animals resulting in significant losses in production. These losses inherently affect the economics of the livestock industry. Since increases in the severity of vector-borne diseases in the southeast Asian region have been reported, investigations of parasitic epidemiology in Thailand will be necessary to improve the existing parasite control strategies for blood parasitic infections. This study aims to investigate incidences of bovine hemoparasites throughout central and northern Thailand by focusing on areas of high-density cattle populations. Methods: Blood parasitic infections among cattle were screened and identified by microscopic examination. Anemia status was then determined by evaluation of the packed cell volume (PCV) of each animal. Furthermore, blood parasites were detected and identified by genus and species-specific primers through the polymerase chain reaction method. Amplicons were subjected to DNA sequencing; thereafter, phylogenetic trees were constructed to determine the genetic diversity and relationships of the parasite in each area. Results: A total of 1,066 blood samples were found to be positive for blood parasitic infections as follows: 13 (1.22%), 389 (36.50%), and 364 (34.15%) for Babesia bovis, Theileria orientalis, and Anaplasma marginale, respectively. Furthermore, multiple hemoparasitic infections in the cattle were detected. The hematocrit results revealed 161 hemoparasitic infected samples from 965 blood samples, all of which exhibiting indications of anemia with no significant differences. Sequence analysis of the identified isolates in this study revealed that B. bovis rap-1, four separate clades of T. orientalis msps, and A. marginale msp4 exhibited considerable sequence similarity to homologous sequences from isolates obtained from other countries. Sequence similarity ranged between 98.57-100%, 83.96-100%, and 97.60-100% for B. bovis rap-1, T. orientalis msps, and A. marginale msp4, respectively. Conclusion: In this study, the analyzed incidence data of cattle hemoparasitic infection in Thailand has provided valuable and basic information for the adaptation of blood-borne parasitic infections control strategies. Moreover, the data obtained from this study would be useful for future effective parasitic disease prevention and surveillance among cattle.

Phylogeography and Prevalence of Hemoparasites (Apicomplexa: Eucoccidiorida) in Galápagos Marine Iguanas, Amblyrhynchus cristatus (Reptilia: Iguanidae).

Parasitism is among the most common forms of coexistence of organisms of different species. Hemoparasites live in the bloodstream of the host where they complete different life-cycle stages. Members of the phylum Apicomplexa constitute a large portion of all hemoparasites infecting reptiles and their parasite transmitting vectors, including arthropods. In this study, we carried out a survey and molecular identification of hemoparasites in blood samples of the iconic Galápagos marine iguana (Amblyrhynchus cristatus). Major island populations of marine iguanas were sampled to examine large-scale biogeographic patterns of parasite diversity and prevalence. Nested PCRs were used to amplify segments of the 18S rRNA-gene of hemoparasites. Furthermore, ticks attached to marine iguanas were collected and analyzed in the same way to assess their potential use as a non-invasive method for the detection of hemoparasites in vertebrate host species. PCR products were sequenced and a phylogenetic analysis was carried out showing the presence of two genetically distinct clusters of hemoparasites, one more commonly distributed than the other one, belonging to the genera Hepatozoon and/or Hemolivia (Apicomplexa: Eucoccidiorida). Overall, 25% of marine iguanas were infected by hemoparasites. However, infection rates varied strongly among particular island populations (from 3.45% to 50%). Although marine iguanas are an extremely mobile species that has colonized all islands in the Galápagos archipelago, parasite occurrence was not related to geographical distance, suggesting that dispersal behavior has a minor role in parasite transmission. On most islands, females tended to have higher infection rates than males, but this relationship was only significant on one island. Overall, ticks and marine iguanas had similar prevalence and diversity of parasites. However, the infection profiles of ticks and their corresponding hosts (marine iguanas) did not mirror one another, indicating that this method cannot be used reliably to assess marine iguana infection status. Interestingly, we found that hemoparasite prevalence in marine iguanas and ticks tended to be positively correlated across islands. Our results indicate that certain populations of marine iguanas may have special mechanisms and adaptations to cope with parasite infection. In addition, other factors such as vector density, anthropogenic-related activities or the immunological state of marine iguanas could potentially affect the striking variation in hemoparasite prevalence across island populations.

Outbreak of follicular conjunctivitis associated with the diagnosis of anaplasmosis in a sheep herd.

OBJECTIVE: To report an outbreak of follicular conjunctivitis in a group of sheep diagnosed with Anaplasma spp., without any other co-infection. ANIMALS STUDIED: In all, 18 animals from a sheep head, males and females, from eight months to four years of age, were assessed for follicular conjunctivitis. PROCEDURES: The procedures performed included general physical and ophthalmological examinations; PCR evaluation for infectious agents; analysis of hematological parameters, microbiological tests of ocular swabs, coproparasitological examination, histopathological examination of conjunctival biopsy. RESULTS: All 18 animals had uni- or bilateral follicular conjunctivitis, and one animal also had unilateral uveitis. The results of microbiological analyzes were negative for Moraxella spp., Staphylococcus spp., and Pseudomonas spp., and PCR analysis results were negative for Chlamydia spp., Mycoplasma spp., and Toxoplasma gondii. Anemia, thrombocytopenia, lymphocytosis, and an inclusion body in some erythrocytes, compatible with Anaplasma and PCR analysis for Anaplasma spp. were positive. CONCLUSION: Anaplasmosis may be associated with follicular conjunctivitis in sheep and should be included in the differential diagnosis list and investigated in cases of conjunctivitis in herds.