Diversity of Anaplasmataceae Transmitted by Ticks (Ixodidae) and the First Molecular Evidence of Anaplasma phagocytophilum and Candidatus Anaplasma boleense in Paraguay.

Anaplasmataceae bacteria are emerging infectious agents transmitted by ticks. The aim of this study was to identify the molecular diversity of this bacterial family in ticks and hosts, both domestic and wild, as well as blood meal sources of free-living ticks in northeastern Paraguay. The bacteria were identified using PCR-HRM, a method optimized for this purpose, while the identification of ticks and their blood meal was performed using conventional PCR. All amplified products were subsequently sequenced. The bacteria detected in the blood hosts included Ehrlichia canis, Anaplasma platys, and Anaplasma phagocytophilum, Candidatus Anaplasma boleense, and Wolbachia spp., which had not been previously reported in the country. Free-living and parasitic ticks on dogs (Canis lupus familiaris) and wild armadillos (Dasypus novemcinctus) were collected and identified as Rhipicephalus sanguineus and Amblyomma spp. The species E. canis, A. platys, A. phagocytophilum, and Ca. A. boleense were detected in domestic dog ticks, and E. canis and A. platys were found for the first time in armadillos and free-living ticks. Blood feeding sources detected in free-living ticks were rodents, humans, armadillos and dogs. Results show a high diversity of tick-borne pathogens circulating among domestic and wild animals in the northeastern region of Paraguay.

Diversity of Ehrlichia spp., Anaplasma spp. and Neorickettsia spp. in vampire bats.

Although bats (Mammalia: Chiroptera) act as natural reservoirs for many zoonotic pathogens around the world, few studies have investigated the occurrence of Anaplasmataceae agents in bats, especially vampire bats. The family Anaplasmataceae (order Rickettsiales) encompasses obligate intracellular bacteria of the genera Anaplasma, Ehrlichia, Neorickettsia, Neoehrlichia, Wolbachia, and Allocryptoplasma. The present study aimed to investigate, using molecular techniques, the presence of species of Anaplasma, Ehrlichia, and Neorickettsia in vampire bats sampled in northern Brazil. Between 2017 and 2019, spleen samples were collected from vampire bats belonging to two species, Desmodus rotundus (n = 228) from the states of Pará (n = 207), Amazonas (n = 1), Roraima (n = 18) and Amapá (n = 3), and Diaemus youngii (n = 1) from Pará. Positivity rates of 5.2% (12/229), 3% (7/229), and 10.9% (25/229) were found in PCR assays for Anaplasma spp. (16S rRNA gene), Ehrlichia spp. (dsb gene) and Neorickettsia spp. (16S rRNA gene), respectively. The present study revealed, for the first time, the occurrence of Anaplasma spp. and different genotypes of Ehrlichia spp. in vampire bats from Brazil. While phylogenetic analyses based on the dsb and ftsZ genes of Ehrlichia and 16S rRNA of Anaplasma spp. revealed phylogenetic proximity of the genotypes detected in vampire bats with Anaplasmataceae agents associated with domestic ruminants, phylogenetic inferences based on the gltA and groEL genes evidenced the occurrence of genotypes apparently exclusive to bats. Neorickettsia sp. phylogenetically associated with N. risticii was also detected in vampire bats sampled in northern Brazil.

Novel Tick-Borne Anaplasmataceae Genotypes in Tropical Birds from the Brazilian Pantanal Wetland.

Despite numerous reports of Anaplasmataceae agents in mammals worldwide, few studies have investigated their occurrence in birds. The present study aimed to investigate the occurrence and molecular identity of Anaplasmataceae agents in birds from the Pantanal wetland, Brazil. Blood samples were collected from 93 different species. After DNA extraction, samples positive for the avian ß-actin gene were subjected to both a multiplex quantitative real-time (q)PCR for Anaplasma and Ehrlichia targeting the groEL gene and to a conventional PCR for Anaplasmataceae agents targeting the 16S rRNA gene. As a result, 37 (7.4%) birds were positive for Anaplasma spp. and 4 (0.8%) for Ehrlichia spp. in the qPCR assay; additionally, 13 (2.6%) were positive for Anaplasmataceae agents in the PCR targeting the 16S rRNA gene. The Ehrlichia 16S rRNA sequences detected in Arundinicola leucocephala, Ramphocelus carbo, and Elaenia albiceps were positioned closely to Ehrlichia sp. Magellanica. Ehrlichia dsb sequences detected in Agelasticus cyanopus and Basileuterus flaveolus grouped with Ehrlichia minasensis. The 16S rRNA genotypes detected in Crax fasciolata, Pitangus sulphuratus and Furnarius leucopus grouped with Candidatus Allocryptoplasma. The 23S-5S genotypes detected in C. fasciolata, Basileuterus flaveolus, and Saltator coerulescens were related to Anaplasma phagocytophilum. In conclusion, novel genotypes of Anaplasma, Ehrlichia, and Candidatus Allocryptoplasma were detected in birds from the Pantanal wetland.

Enfermedades transmitidas por vectores en gatos: una mirada molecular en ambientes urbanos de Medellín, Colombia / Vector-borne diseases in cats: a molecular look at urban environments in Medellin, Colombia

RESUMEN Los microorganismos hemotrópicos en felinos son agentes infecciosos que varían desde nematodos, protozoos y bacterias. El presente estudio retrospectivo tiene como objetivo evaluar la frecuencia de agentes hemotrópicos mediante qPCR de las bases de datos de 1.418 felinos en Medellín entre julio de 2021 y marzo de 2022, periodo en el que se evidencia una frecuencia del 70%, con un número de animales infectados con uno, dos o tres agentes del 56%, 14%, y 2,3%, respectivamente. La frecuencia para cada uno de los agentes es: Rickettsia spp. 0,21%, Babesia spp. 0,35%, Ehrlichia spp. 0,49%, Dirolifaria spp. 0,64%, Anaplasma spp 0,7%, Hepatozoon spp. 5,4%, Mycoplasma spp. 24,4% y Bartonella spp. 37,9%. Las coinfecciones evidenciadas de dos agentes hemotrópicos son: Bartonella spp. y Mycoplasma spp. 7,9%, Bartonella spp. y Hepatozoon spp. 2,1%, Mycoplasma spp. y Hepatozoon spp. 2% y Ehrlichia spp. y Anaplasma spp. 0,5%. De los 15 hemogramas de felinos infectados, 11 de ellos tienen hemogramas sin alteraciones significativas. Dos de los felinos positivos evidencian anemia moderada y severa y reticu-locitos de 0,9% y 0,4%, respectivamente. Solo un individuo positivo para Mycoplasma spp. presenta trombocitopenia y tres plaquetas en limites inferiores. Se concluye que la PCR es la prueba más confiable para el diagnóstico de agentes hemotrópicos.

ABSTRACT Hemotropic microorganisms in felines are infectious agents that vary from nematodes, protozoa, and bacteria. The objective of this retrospective study is to evaluate the frequency of hemotropic agents by means of qPCR from the databases of 1,418 felines in the city of Medellín between July 2021 and March 2022, where a frequency of 70% is evidenced, with several infected animals, with one, two, or three agents of 56%, 14%, and 2.3%, respectively. With a frequency for each of the agents of: Rickettsia spp. 0.21%, Babesia spp. 0.35%, Ehrlichia spp. 0.49%, Dirolifaria spp. 0.64%, Anaplasma spp. 0.7%, Hepatozoon spp. 5.4%, Mycoplasma spp. 24.4%, and Bartonella spp. 37.9% The evidenced coinfections of two hemotropic agents is: Bartonella spp. and Mycoplasma spp. 7.9%, Bartonella spp. and Hepatozoon spp. 2.1%, Mycoplasma spp. and Hepatozoon spp. 2% and Ehrlichia spp. and Anaplasma spp. 0.5%. Of the 15 blood counts from infected cats, 11 of them had blood counts without significant changes. Two of the positive cats show moderate and severe anemia, and reticulocytes of 0.9% and 0.4%, respectively. Only one individual positive for Mycoplasma spp. presented thrombocytopenia, and three platelets in lower limits. It is concluded that PCR is the most reliable test for the diagnosis of hemotropic agents.

First report of pediatric ehrlichiosis in Mexico.

BACKGROUND: Ehrlichia chaffeensis is responsible for most cases of human ehrlichiosis, an acute febrile tick-borne disease. This clinical entity is more commonly reported in adults from the United States. Therefore, it is of special interest to characterize this disease in children, given that very few cases in children have been reported outside of this country. CASE REPORT: We describe the case of a 15-year-old female from northeastern Mexico with a five-day history of myalgias, arthralgias, fever, abdominal pain, rash, and somnolence. The possibility of tick-borne disease was suspected considering that she lived with three tick-infested dogs that had recently died and a neighbor with similar symptoms who deteriorated rapidly and died a week earlier. Ehrlichia spp. was detected in blood samples by polymerase chain reaction. The patient completed a seven-day course of doxycycline and was discharged with complete resolution of symptoms. CONCLUSIONS: This case is the first report of ehrlichiosis in a pediatric patient in Mexico, illustrating the importance of considering tick-borne diseases as a differential diagnosis in patients with rash, fever, and altered level of consciousness. This initial clinical presentation may be indistinct from other conditions such as dengue, meningococcemia, and multisystem inflammatory syndrome in children (MIS-C), among others.

INTRODUCCIÓN: Ehrlichia chaffeensis es responsable de la mayoría de los casos de ehrlichiosis humana, una enfermedad febril aguda transmitida por garrapatas. Esta entidad clínica se reporta con mayor frecuencia en adultos de Estados Unidos. Por lo tanto, es de especial interés caracterizarla en niños, dado que se han reportado muy pocos casos en niños fuera de este país. CASO CLÍNICO: Se describe el caso de una paciente de sexo femenino de 15 años, originaria y residente del noreste de México con una historia de cinco días de mialgias, artralgias, fiebre, dolor abdominal, erupción cutánea y somnolencia. Se sospechó la posibilidad de una enfermedad transmitida por garrapatas considerando que convivió con tres perros infestados de garrapatas que habían muerto recientemente y una vecina con síntomas similares, quien se deterioró rápidamente y murió una semana antes. Ehrlichia spp. se detectó en una muestra sérica mediante reacción en cadena de la polimerasa. La paciente completó un curso de siete días de doxiciclina y fue dada de alta con resolución de los síntomas. CONCLUSIONES: Este caso es el primer reporte de ehrlichiosis en un paciente pediátrico en México que ilustra la importancia de considerar enfermedades transmitidas por garrapatas dentro del diagnóstico diferencial de pacientes con exantema, fiebre y alteración del estado de conciencia. Esta presentación clínica inicial puede ser indistinguible de otras entidades como dengue, meningococcemia y síndrome multisistémico inflamatorio, entre otras.

Molecular detection and characterization of vector-borne agents in common opossums (Didelphis marsupialis) from northeastern Brazil.

Opossums are synanthropic marsupials able to interchange among wild, peri­urban and urban environments, playing an epidemiologically important role as hosts for emerging pathogens and ectoparasites of relevance in public health. The present study aimed to detect and molecularly characterize vector-borne agents in a population of common opossums (Didelphis marsupialis) from the Island of São Luís do Maranhão, northeastern Brazil. Of the 45 animals analyzed, one (2.22%) was positive in the nested PCR assay based on the 18S rRNA gene of piroplasmids. The obtained sequence was phylogenetically positioned in a clade containing sequences of Babesia sp. previously detected in Didelphis aurita, Didelphis albiventris and associated ticks from Brazil. Eight (17.77%) samples were positive in PCR for Ehrlichia spp. based on the dsb gene; four samples were sequenced and positioned into a new clade, sister to E. minasensis and Ehrlichia sp. clade detected in Superorder Xenarthra mammals. No samples tested positive in the screening PCR assays based on the 16S rRNA gene of Anaplasma spp. Two samples were positive in the qPCR for Bartonella spp. based on the nuoG gene. Seven animals (15.56%) were positive in the nPCR based on the 16S rRNA gene of hemoplasmas. Of these, three were positive in a PCR based on the 23S rRNA gene. The phylogenies based on both 16S rRNA and 23S rRNA genes corroborated to each other and positioned the sequences in the same clade of hemoplasmas previously detected in D. aurita and D. albiventris sampled in Brazil. Finally, three (6.66%) animals were positive in the PCR for Hepatozoon spp.; the obtained 18S rRNA sequence was positioned into the H. felis clade.The present study showed, for the first time, the circulation of piroplasmids, Hepatozoon spp., Ehrlichia spp., hemoplasmas and Bartonella spp. in D. marsupialis sampled in northeastern Brazil, with description of putative novel genotypes of Ehrlichia and Hepatozoon and copositivity by different vector-borne agents. The present work consolidates the "South American Marsupialia" piroplasmid clade, adding one more genotype of Babesia sp. to this clade.

First report of pediatric ehrlichiosis in Mexico / Primer reporte de ehrlichiosis pediátrica en México

Abstract Background: Ehrlichia chaffeensis is responsible for most cases of human ehrlichiosis, an acute febrile tick-borne disease. This clinical entity is more commonly reported in adults from the United States. Therefore, it is of special interest to characterize this disease in children, given that very few cases in children have been reported outside of this country. Case report: We describe the case of a 15-year-old female from northeastern Mexico with a five-day history of myalgias, arthralgias, fever, abdominal pain, rash, and somnolence. The possibility of tick-borne disease was suspected considering that she lived with three tick-infested dogs that had recently died and a neighbor with similar symptoms who deteriorated rapidly and died a week earlier. Ehrlichia spp. was detected in blood samples by polymerase chain reaction. The patient completed a seven-day course of doxycycline and was discharged with complete resolution of symptoms. Conclusions: This case is the first report of ehrlichiosis in a pediatric patient in Mexico, illustrating the importance of considering tick-borne diseases as a differential diagnosis in patients with rash, fever, and altered level of consciousness. This initial clinical presentation may be indistinct from other conditions such as dengue, meningococcemia, and multisystem inflammatory syndrome in children (MIS-C), among others.

Resumen Introducción: Ehrlichia chaffeensis es responsable de la mayoría de los casos de ehrlichiosis humana, una enfermedad febril aguda transmitida por garrapatas. Esta entidad clínica se reporta con mayor frecuencia en adultos de Estados Unidos. Por lo tanto, es de especial interés caracterizarla en niños, dado que se han reportado muy pocos casos en niños fuera de este país. Caso clínico: Se describe el caso de una paciente de sexo femenino de 15 años, originaria y residente del noreste de México con una historia de cinco días de mialgias, artralgias, fiebre, dolor abdominal, erupción cutánea y somnolencia. Se sospechó la posibilidad de una enfermedad transmitida por garrapatas considerando que convivió con tres perros infestados de garrapatas que habían muerto recientemente y una vecina con síntomas similares, quien se deterioró rápidamente y murió una semana antes. Ehrlichia spp. se detectó en una muestra sérica mediante reacción en cadena de la polimerasa. La paciente completó un curso de siete días de doxiciclina y fue dada de alta con resolución de los síntomas. Conclusiones: Este caso es el primer reporte de ehrlichiosis en un paciente pediátrico en México que ilustra la importancia de considerar enfermedades transmitidas por garrapatas dentro del diagnóstico diferencial de pacientes con exantema, fiebre y alteración del estado de conciencia. Esta presentación clínica inicial puede ser indistinguible de otras entidades como dengue, meningococcemia y síndrome multisistémico inflamatorio, entre otras.

Patógenos transmitidos por vetores na reprodução canina / Vector-borne pathogens in canine reproduction

O artigo apresenta uma revisão sobre os efeitos reprodutivos da infecção pelos principais patógenos transmitidos por vetores que acometem cães, Leishmania spp., Babesia spp., Ehrlichia spp., Anaplasma spp. e Trypanosoma spp. São discutidas a patologia genital, fertilidade masculina e feminina, transmissão venérea, transmissão vertical, transmissão horizontal mãe-filho e sua relevância na epidemiologia das doenças.(AU)

The article presents a review of the reproductive effects of infection by main vector-borne pathogens of dogs, Leishmania spp., Babesia spp., Ehrlichia spp., Anaplasma spp. and Trypanosoma spp. Genital pathology, male and female fertility, venereal transmission, vertical transmission, horizontal mother-offspring transmission and their relevance to the epidemiology of the diseases are discussed.(AU)

Diagnóstico hematológico de coinfecção por Ehrlichia spp. e Hepatozoon canis em cães no semiárido da Paraíba / Hematogical diagnosis of coinfection by Ehrlichia spp. and Hepatozoon canis in dogs in the semi-arid of Paraiba

Dentre as hemoparasitoses com maior frequência de transmissão por carrapatos pode-se destacar a Erliquiose, a Hepatozoonose, a Anaplasmose e a Babesiose. A coinfecção por esses agentes são frequentes na rotina clínica; no entanto, poucas são as descrições sobre a ocorrência simultânea desses agentes em hemogramas. Sendo assim, o objetivo deste trabalho foi relatar os casos de quatro cães diagnosticados com coinfecção por Ehrlichia spp. e Hepatozoon canis, os quais foram atendidos no Hospital Veterinário Adílio Santos de Azevedo (HV-ASA) do Instituto Federal de Educação, Ciência e Tecnologia da Paraíba (IFPB), campus Sousa. Durante a avaliação clínica, relatou-se que os pacientes apresentavam, de maneira em comum, oligodipsia e hiporexia, além de histórico de presença de carrapatos. Em seguida, foram realizados hemogramas, que revelaram que todos os animais apresentavam anemia leve a moderada. Além disso, notou-se que o animal 1 estava com trombocitopenia e os animais 2 e 3, neutrofilia. Na pesquisa de hemoparasitas em esfregaço sanguíneo, observou-se coinfecção por H. canis e E. spp. nos quatro animais. Desta forma, este relato demonstrou a importância do uso do hemograma e da pesquisa dos agentes causadores das hemoparasitoses, através do esfregaço sanguíneo. Ressalte-se que o esfregaço sanguíneo, embora seja um método inespecífico e de baixa sensibilidade, pode e deve ser adotado como rotina, por ser uma técnica simples, rápida, barata, pouco invasiva e de diagnóstico definitivo.

Among the hemoparasitoses with the highest frequency of transmission by ticks, ehrlichiosis, hepatozoonosis, anaplasmosis, and babesiosis may be highlighted. The coinfection with these agents is frequent in clinical routine; however, there are few descriptions of the simultaneous occurrence of these agents in blood counts. Therefore, this work aimed to report the cases of four dogs diagnosed with Ehrlichia spp. and Hepatozoon canis, which were treated at the Adílio Santos de Azevedo Veterinary Hospital (HV-ASA), of the Federal Institute of Education, Science, and Technology of Paraíba (IFPB), Sousa campus. During the clinical evaluation, it was reported that the patients had oligodipsia and hyporexia in common, in addition to a history of tick presence. Then, blood counts were performed, which revealed that all animals had mild to moderate anemia. In addition, it was noted that animal 1 had thrombocytopenia and animals 2 and 3 had neutrophilia. In the investigation of hemoparasites in blood smears, co-infection with H. canis and E. spp. was observed in the four animals. Thus, this report demonstrated the importance of using the blood count and researching the causative agents of hemoparasitosis, through the blood smear. It should be noted that the blood smear, although it is a non-specific and low-sensitivity method, can and should be adopted as a routine, as it is a simple, fast, inexpensive, minimally invasive, and definitive diagnostic technique.

Novel Ehrlichia and Hepatozoon genotypes in white-eared opossums (Didelphis albiventris) and associated ticks from Brazil.

White-eared opossums (Didelphis albiventris) are well adapted to anthropized areas. The increased contact with domestic animals and humans mediates the transmission of arthropod-borne pathogens. Despite the worldwide occurrence of tick-borne Anaplasmataceae and Hepatozoidae species in a variety of vertebrates, few studies reported serological evidence or molecular detection of theses agentes in marsupials. Up to now, while Ehrlichia/Anaplasma spp. have only been detected in marsupials from Brazil, Hepatozoon spp. have been reported in marsupials from Chile, Australia and Brazil. The present work aimed to investigate, using molecular techniques and blood smear analysis, the presence of Ehrlichia spp., Anaplasma spp., and Hepatozoon sp. in the blood and ticks collected from D. albiventris in urban forest fragments from midwestern Brazil. Between May and December 2017, 43 D. albiventris (27 males and 16 females) were captured for blood and tick collection in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil. Ticks (46 Amblyomma dubitatum nymphs and 24 Amblyomma spp. larvae) were collected from 14 out 43 (32.5%) of the white-eared opossums. Panoptic-stained blood smears were performed using peripheral blood (tail tip) of the captured opossums. DNA extracted from blood and tick samples were subjected to PCR/qPCR assays for Anaplasmataceae agents (rrs, gltA, groEL, sodB, and dsb genes, and 23S-5S intergenic region) and Hepatozoon spp. (18S rRNA gene), followed by Sanger sequencing, BLASTn and phylogenetic analyses. An inclusion resembling Ehrlichia morulae was found in a white-eared opossum's monocyte from a blood smear stained with Panoptic. Five (11.63% [5/43]) white-eared opossums' blood samples and 7 (25% [7/28]) tick samples (2 pools of Amblyomma spp. larvae and 5 pools of A. dubitatum nymphs) were positive for Anaplasmataceae via a PCR assay targeting the conserved rrs gene. Phylogenetic analysis based on the rrs gene positioned three sequences obtained from opossums and ticks together as a subclade within the Ehrlichia canis clade. However, all samples were negative in a qPCR assay specific for E. canis based on the dsb gene. Phylogenetic analyses positioned the gltA and 23S-5S ITS sequences obtained from opossums' blood samples in a separate clade from the other validated Ehrlichia species. One (2.3% [1/43]) opossum blood sample was positive for the 18S rRNA gene of Hepatozoon sp. The phylogenetic analysis positioned the Hepatozoon sp. sequence obtained from a D. albiventris specimen in a clade with a sequence previously detected in a black storm petrel (Oceanodroma melania) from Mexico. All the other sequences of Hepatozoon sp. previously detected in marsupials from Brazil were positioned in a separated clade. The present work showed the occurrence of putative novel genotypes of Ehrlichia sp. and Hepatozoon sp. in white-eared opossums and associated A. dubitatum ticks from midwestern Brazil.

Current distribution of selected canine vector-borne diseases in domestic dogs from Barranquilla and Puerto Colombia, Atlántico, Colombia.

BACKGROUND: Climate change, the increase of travel with infected animals from endemic areas, the introduction of new vectors in these areas and environmental changes caused by human activity, among other factors, have contributed to the establishment and increase of canine vector-borne diseases (CVBDs), several of which are zoonotic and pose a risk to the human population. In Colombia, there are very few studies that address the prevalence of these diseases. The objective of this study was to update the prevalence of cardiopulmonary dirofilariosis, anaplasmosis, ehrlichiosis and Lyme borreliosis in dogs in Barranquilla and Puerto Colombia, areas of northern Colombia. MATERIALS AND METHODS: The present study included 354 dogs presented to veterinary clinics for routine health examination and foundations for stray dogs between November 2016 and July 2018. RESULTS: The percentage of dogs positive for Ehrlichia spp. was 61.86%, followed by 22.03% for Anaplasma spp., 11.30% positive for Dirofilaria immitis antigens and 0.56% positive for Borrelia burgdorferi. In addition, several dogs positive for antibodies against two or more infectious diseases were found. Higher seroprevalences were documented in outdoor dogs compared to indoor-housed dogs. CONCLUSION: These results suggest that veterinarians should routinely implement prophylactic programmes for these CVBDs, particularly for dogs that reside outdoors.

Trypanosoma cruzi co-infections with other vector borne diseases are frequent in dogs from the pacific coast of Ecuador.

Dogs are a reservoir for Chagas disease, caused by Trypanosoma cruzi (T. cruzi), and other companion vector-borne diseases, including ehrlichiosis (Ehrlichia canis and Ehrlichia ewingii), anaplasmosis (Anaplasma phagocytophilum and Anaplasma platys), dirofilariasis (Dirofilaria immitis) and Lyme disease (Borrelia burgdorferi). This study has two key objectives: 1) to determine seroreactivity against T. cruzi in dogs from the town of Colón, in Portoviejo city, in the central coast of Ecuador; and 2) to establish the coinfection frequency of other companion vector-borne diseases in dogs positive for T. cruzi. Antibodies against T. cruzi were detected using two enzyme-linked immunosorbent assays. Diagnostic consensus between ELISA tests was established using the Cohen's Kappa coefficient. Other haemoparasitic diseases were detected using the IDEXX SNAP® 4Dx® kit in dogs previously diagnosed as T. cruzi-seropositive. From 84 dogs sampled, 57.14% (48/84) tested positive for T. cruzi. Co-infection analysis of 25 dogs positive for T. cruzi revealed antibodies also against Ehrlichia spp. (48%), Anaplasma spp. (28%), and Dirofilaria immitis (12%). These results provide a novel perspective regarding the status of these pathogens which co-infect dogs in Colón. Since all these pathogens are zoonotic, our findings should warn regional health authorities to implement sanitary programs, to better prevent and control vectors associated to these pathogens. On the other hand, human and veterinarian doctors, should consider that patients with a cardiac infection condition could be suffering co-infections with two or more vector transmitted pathogens.

Tick-borne pathogens in carthorses from Foz do Iguaçu City, Paraná State, southern Brazil: A tri-border area of Brazil, Paraguay and Argentina.

Tick-borne diseases (TBD) constitute an important group of illness affecting animals and humans worldwide. In Brazil, carthorses are frequently exposed to ticks and tick-borne pathogens, leading to impairment of horse performance and imposing restrictions by the international veterinary authorities for the importation of horses. Accordingly, this study has aimed to i) determine the prevalence of the TBD agents Theileria equi, Babesia caballi, Ehrlichia spp., and hemotropic mycoplasmas in carthorses, ii) identify the tick species parasitizing the animals, and iii) determine factors associated with exposure/infection in Foz do Iguaçu City, Parana state, southern Brazil. A total of 103 carthorses were screened for anti-T. equi and anti-Ehrlichia spp. antibodies by indirect fluorescent antibody assays (IFA). Samples were also tested by PCR assays targeting the 18S rRNA gene of T. equi and B. caballi, and 16S rRNA gene of hemoplasmas. Additionally, PCR assays targeting the 16S rRNA, disulfide bond formation protein (dsb) and tandem repeat proteins 36 (trp36) genes of Ehrlichia spp. were also performed. Antibodies to T. equi and Ehrlichia spp. were detected in 43/103 (41.75%; 95% CI: 32.10-51.88%) and 5/103 (4.85%; 95% CI: 1.59-10.97%) horses by IFA, respectively. DNA of T. equi and B. caballi were found in 25/103 (24.27%; 95% CI: 16.36-33.71%) and 10/103 (9.71%; 95% CI: 4.75-17.13%) carthorses, respectively, and all tested negative for Ehrlichia spp. and hemoplasmas. All sequences showed ≥99% identity with multiple T. equi and B. caballi 18S rRNA gene sequences deposited in GenBank. Overall, 191 Dermacentor nitens ticks were collected from 25/103 (24.27%) animals. Carthorses older than 5 years were more likely to be positive for T. equi (p < 0.05). In conclusion, equine piroplasmosis agents are highly prevalent in carthorses from Foz do Iguaçu City. The low prevalence of Ehrlichia spp. found may be due to the absence of Amblyomma ticks infesting animals, which should be further investigated.

Seroprevalence and Current Infections of Canine Vector-Borne Diseases in Costa Rica.

Domestic dogs may carry several vector-borne pathogens, including zoonotic agents, especially in tropical regions like Central America. The epidemiology of these pathogens is prone to change due to urbanization, trade and travel as well as climate change, necessitating repeated monitoring. This study aims to present a comprehensive picture of canine vector-borne diseases in Costa Rica, combining data on seroprevalence with molecular species identification of the causative pathogens. In this survey, 294 dogs from all seven provinces of Costa Rica were included. After a clinical examination, diagnostic blood samples were analyzed with regard to packed cell volume (PCV) and presence of microfilaria. Serum samples were tested for antibodies against Ehrlichia spp., Anaplasma spp., Babesia spp., Borrelia burgdorferi sensu lato (s.l.) as well as antigen of Dirofilaria immitis. Seropositive and microfilaremic blood samples were analyzed by PCR to detect current infections and identify the pathogen species. Overall, 45.24% (133/294, 95% CI: 39.45-51.11%) of dogs were seropositive for at least one of the tested pathogens. Seroprevalence was highest for Ehrlichia spp. (39.46%, 116/294, 95% CI: 33.83-45.29%), followed by Babesia spp. (23.13%, 68/294, 95% CI: 18.43-28.38%), Anaplasma spp. (13.27%, 39/294, 95% CI: 9.61-17.69%), and B. burgdorferi s.l. (0.34%, 1/294, 95% CI: 0.01-1.88%). Generalized linear mixed models indicated a significant association of Ehrlichia-, Anaplasma- and Babesia-seropositivity, as well as a significant effect of age and breed on Ehrlichia-seropositivity. Furthermore, a statistically significant negative effect of Ehrlichia-, Anaplasma-, and Babesia-seropositivity on PCV was found. Regarding current infections, Ehrlichia canis DNA was detected in 51.72% (60/116, 95% CI: 42.26-61.10%) of Ehrlichia-seropositive dogs, while Ehrlichia ewingii and Ehrlichia chaffeensis were not detected. Furthermore, 10.26% (4/39, 95% CI: 2.87-24.22%) of Anaplasma-seropositive dogs were coinfected with Anaplasma phagocytophilum and Anaplasma platys, while one animal (2.56%, 95% CI: 0.65-13.48%) was infected with A. phagocytophilum only. Among Babesia-seropositive dogs, Babesia vogeli and Hepatozoon canis were detected in one animal each (1.47%, 1/68, 95% CI: 0.04-7.92%). Dirofilaria immitis antigen was detected in 4.42% (13/294, 95% CI: 2.38-7.44%) of dogs. In microfilaremic animals, D. immitis as well as Acanthocheilonema reconditum infections were identified. This survey demonstrates that canine vector-borne pathogens, including zoonotic agents like A. phagocytophilum and D. immitis, are widespread in Costa Rica. Thus, protection of dogs from disease-transmitting vectors is recommended from an animal welfare as well as public health perspective.

Seroprevalence and current infections of canine vector-borne diseases in Nicaragua.

BACKGROUND: Vector-borne diseases constitute a major problem for veterinary and public health, especially in tropical regions like Central America. Domestic dogs may be infected with several vector-borne pathogens of zoonotic relevance, which may also severely compromise canine health. METHODS: To assess the prevalence of canine vector-borne diseases in Nicaragua, 329 dogs from seven cities, which were presented to the veterinarian for various reasons, were included in this study. Dogs were examined clinically and diagnostic blood samples were taken for analysis of packed cell volume (PCV) and presence of microfilariae as well as antigen of Dirofilaria immitis and antibodies to Ehrlichia spp., Anaplasma spp. and Borrelia burgdorferi (sensu lato) by use of a commercially available rapid ELISA. To detect current infections, specific PCRs for the detection of E. canis, A. platys and A. phagocytophilum were carried out on blood samples of the respective seropositive dogs. Microfilaremic blood samples, as well as D. immitis antigen positive samples were further subjected to PCR and subsequent sequencing for filarial species identification. RESULTS: Antibodies against Ehrlichia spp. were present in 62.9% of dogs, while Anaplasma spp. seroprevalence was 28.6%. Antibodies against species of both genera were detected in 24.9% of dogs. Borrelia burgdorferi (s.l.) antibodies were not detected. Dirofilaria immitis antigen was present in six animals (1.8%), two of which also showed D. immitis microfilariae in buffy coat. In addition to D. immitis, Acanthocheilonema reconditum was identified by PCR and sequencing in two of four additional microfilaremic blood samples, which were tested negative for D. immitis antigen. Current E. canis infections as defined by DNA detection were present in 58.5% of Ehrlichia-seropositive dogs, while 5.3% of Anaplasma-seropositive dogs were PCR-positive for A. platys, 2.2% for A. phagocytophilum and 16.0% for both Anaplasma species. Current E. canis infection had a statistically significant negative impact on PCV, whereas no relationship between infection status and clinical signs of disease could be observed. CONCLUSIONS: These results indicate that canine vector-borne diseases are widespread in Nicaragua and that dogs may constitute a reservoir for human infection with E. canis, A. phagocytophilum and D. immitis. Thus, the use of repellents or acaricides to protect dogs from vector-borne diseases is strongly recommended.

Antibodies against canine distemper virus, parvovirus and Ehrlichia spp. in wild captive carnivores in midwestern Brazil / Anticorpos contra o vírus da cinomose canina, parvovírus e Ehrlichia spp. em carnívoros selvagens cativos no centro-oeste do Brasil

The occurrence of antibodies against canine distemper virus (CDV), parvovirus and Ehrlichia spp. in wild captive carnivores was evaluated in a zoological park in midwestern Brazil. Serum samples were collected between 2007 and 2014 from 45 carnivores. Antibodies were evaluated by virus neutralization assay for CDV, hemagglutination inhibition test for parvovirus, indirect immunofluorescent and Enzyme-linked immunosorbent assay for Ehrlichia spp. Antibodies against CDV and parvovirus were detected in 75% of Canidae and Felidae. Procyonidae were negative for CDV, although one Mustelidae was positive. TwoCanidae presented antibodies reactive to E. canis antigens. The high antibodies rates to CDV and parvovirus suggest the contact with both pathogens, however since no clinical history of disease are registered in the Zoo-UFMT, we can presume that carnivores have responded satisfactorily against the antigens. The low serological rates observed against Ehrlichia spp. may be resulted to the low occurrence of ticks among carnivores.(AU)

A ocorrência de anticorpos contra o vírus da cinomose canina (CDV), parvovírus e Ehrlichia spp. em carnívoros selvagens em cativeiro foi avaliada em um parque zoológico do centro oeste do Brasil. As amostras de soro foram coletadas entre 2007 e 2014 de 45 carnívoros. Os anticorpos foram avaliados por ensaio de neutralização de vírus para CDV, teste de inibição de hemaglutinação para parvovírus, imunofluorescência indireta e ensaio imunoenzimático ligado à enzima para Ehrlichia spp. Anticorpos contra CDV e parvovírus foram detectados em 75% de canídeos e felídeos. Procionídeos foram negativos para CDV, embora um mustelídeo fora positivo. Dois canídeos apresentaram anticorpos reativos aos antígenos de E. canis. As altas taxas de anticorpos para CDV e parvovírus sugerem o contato com ambos os patógenos, entretanto desde que nenhuma história clínica de doença está registrada no Zoo-UFMT, podemos presumir que os carnívoros têm respondido satisfatoriamente contra os antígenos. As baixas taxas serológicas observadas contra Ehrlichia spp. pode ser resultado da baixa ocorrência de carrapatos entre os carnívoros.(AU)

Antibodies against canine distemper virus, parvovirus and Ehrlichia spp. in wild captive carnivores in midwestern Brazil / Anticorpos contra o vírus da cinomose canina, parvovírus e Ehrlichia spp. em carnívoros selvagens cativos no centro-oeste do Brasil

The occurrence of antibodies against canine distemper virus (CDV), parvovirus and Ehrlichia spp. in wild captive carnivores was evaluated in a zoological park in midwestern Brazil. Serum samples were collected between 2007 and 2014 from 45 carnivores. Antibodies were evaluated by virus neutralization assay for CDV, hemagglutination inhibition test for parvovirus, indirect immunofluorescent and Enzyme-linked immunosorbent assay for Ehrlichia spp. Antibodies against CDV and parvovirus were detected in 75% of Canidae and Felidae. Procyonidae were negative for CDV, although one Mustelidae was positive. TwoCanidae presented antibodies reactive to E. canis antigens. The high antibodies rates to CDV and parvovirus suggest the contact with both pathogens, however since no clinical history of disease are registered in the Zoo-UFMT, we can presume that carnivores have responded satisfactorily against the antigens. The low serological rates observed against Ehrlichia spp. may be resulted to the low occurrence of ticks among carnivores.(AU)

A ocorrência de anticorpos contra o vírus da cinomose canina (CDV), parvovírus e Ehrlichia spp. em carnívoros selvagens em cativeiro foi avaliada em um parque zoológico do centro oeste do Brasil. As amostras de soro foram coletadas entre 2007 e 2014 de 45 carnívoros. Os anticorpos foram avaliados por ensaio de neutralização de vírus para CDV, teste de inibição de hemaglutinação para parvovírus, imunofluorescência indireta e ensaio imunoenzimático ligado à enzima para Ehrlichia spp. Anticorpos contra CDV e parvovírus foram detectados em 75% de canídeos e felídeos. Procionídeos foram negativos para CDV, embora um mustelídeo fora positivo. Dois canídeos apresentaram anticorpos reativos aos antígenos de E. canis. As altas taxas de anticorpos para CDV e parvovírus sugerem o contato com ambos os patógenos, entretanto desde que nenhuma história clínica de doença está registrada no Zoo-UFMT, podemos presumir que os carnívoros têm respondido satisfatoriamente contra os antígenos. As baixas taxas serológicas observadas contra Ehrlichia spp. pode ser resultado da baixa ocorrência de carrapatos entre os carnívoros.(AU)

Infection by Mycoplasma spp., feline immunodeficiency virus and feline leukemia virus in cats from an area endemic for visceral leishmaniasis.

BACKGROUND: Visceral leishmaniasis (VL) has been increasingly recognized in cats living in areas endemic for the disease. Co-infection with Leishmania infantum and other infectious agents is well established in dogs. However, for cats, data on co-infections with L. infantum and other infectious agents are still sparse. The aim of this study was to identify the prevalence of vector-borne pathogens, Mycoplasma spp., feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) in cats from an area endemic for VL in southeastern Brazil. RESULTS: Of the 90 cats, eight (8.9%) were infected with Mycoplasma spp., five (5.5%) were FIV- positive and one (1.1%) was FeLV-positive. Co-infection with L. infantum and at least one other infectious agent was found in 9/50 (18.0%; CI: 8.6-31.4%) cats. In Group 1 (cats infected naturally by L. infantum), 4/50 (8.0%) cats were positive for FIV, 4/50 (8%) for Mycoplasma spp. and 1/50 (2.0%) was co-infected with FeLV and Mycoplasma spp. In Group 2 (cats non-infected with L. infantum), 2/40 (5.0%) cats were infected with Mycoplasma spp. and 1/40 (2.5%) was co-infected with FIV and Mycoplasma spp. All cats were negative for Ehrlichia spp., Babesia spp. and Anaplasma platys. CONCLUSION: A low prevalence of co-infection in Leishmania-infected and non-infected cats was found. Co-infections with Leishmania and vector-borne diseases in cats are not common in this area endemic for VL in Brazil.

Ehrlichia canis and Rickettsia spp. in dogs from urban areas in Paraiba state, northeastern Brazil / Ehrlichia canis e Rickettsia spp. em cães de áreas urbanas da Paraíba, nordeste do Brasil

Abstract The aims of our study was to identify Ehrlichia canis and antibodies against Rickettsia spp. belonging to the spotted fever group (SFG) in dogs sampled from Paraiba state, northeastern Brazil. Blood and serum samples collected by convenience from dogs in urban areas of five municipalities were analyzed by real-time PCR for the detection of E. canis DNA and by immunofluorescence assay test (IFAT) for the identification of antibodies against Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii and R. rhipicephali antigens. E. canis DNA was detected in 8.9% (64/719) of the blood samples, whereas 5.63% (43/763) of the serum samples were positive for at least one of the Rickettsia antigens tested by IFAT. This study showed for the first time the occurrence of E. canis and suggested the circulation of SFG Rickettsia in dogs in the study region of Paraiba state, northeastern Brazil.

Resumo Os objetivos do nosso estudo foram identificar Ehrlichia canis e anticorpos contra Rickettsia spp. pertencentes ao Grupo da Febre Maculosa (GFM) em cães amostrados no estado da Paraíba, nordeste do Brasil. As amostras de sangue e soro, coletados por conveniência, de cães em áreas urbanas de cinco municípios foram analisadas por PCR em tempo real para a detecção de DNA de E. canis e pela Reação de Imunofluorescência Indireta (RIFI) para identificação de anticorpos contra Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii e R. rhipicephali. O DNA de E. canis foi detectado em 8,9% (64/719) das amostras de sangue, enquanto que 5,63% (43/763) das amostras de soro foram positivas para pelo menos um dos antígenos de Rickettsia testados por RIFI. Este estudo mostrou pela primeira vez a ocorrência de E. canis e sugere a circulação de Rickettsia do GFM em cães na região em estudo do estado da Paraíba, Nordeste do Brasil.

Ehrlichia canis and Rickettsia spp. in dogs from urban areas in Paraiba state, northeastern Brazil / Ehrlichia canis e Rickettsia spp. em cães de áreas urbanas da Paraíba, nordeste do Brasil

The aims of our study was to identify Ehrlichia canis and antibodies against Rickettsia spp. belonging to the spotted fever group (SFG) in dogs sampled from Paraiba state, northeastern Brazil. Blood and serum samples collected by convenience from dogs in urban areas of five municipalities were analyzed by real-time PCR for the detection of E. canis DNA and by immunofluorescence assay test (IFAT) for the identification of antibodies against Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii and R. rhipicephali antigens. E. canis DNA was detected in 8.9% (64/719) of the blood samples, whereas 5.63% (43/763) of the serum samples were positive for at least one of the Rickettsia antigens tested by IFAT. This study showed for the first time the occurrence of E. canis and suggested the circulation of SFG Rickettsia in dogs in the study region of Paraiba state, northeastern Brazil.(AU)

Os objetivos do nosso estudo foram identificar Ehrlichia canis e anticorpos contra Rickettsia spp. pertencentes ao Grupo da Febre Maculosa (GFM) em cães amostrados no estado da Paraíba, nordeste do Brasil. As amostras de sangue e soro, coletados por conveniência, de cães em áreas urbanas de cinco municípios foram analisadas por PCR em tempo real para a detecção de DNA de E. canis e pela Reação de Imunofluorescência Indireta (RIFI) para identificação de anticorpos contra Rickettsia rickettsii, R. felis, R. parkeri, R. amblyommii e R. rhipicephali. O DNA de E. canis foi detectado em 8,9% (64/719) das amostras de sangue, enquanto que 5,63% (43/763) das amostras de soro foram positivas para pelo menos um dos antígenos de Rickettsia testados por RIFI. Este estudo mostrou pela primeira vez a ocorrência de E. canis e sugere a circulação de Rickettsia do GFM em cães na região em estudo do estado da Paraíba, Nordeste do Brasil.(AU)