Molecular detection of Anaplasma species in dogs in Colombia

Abstract

Anaplasma platys and A. phagocytophilum are tick-borne pathogens that parasitize platelets and neutrophils, respectively, of humans and animals. The former is the etiological agent of canine cyclic thrombocytopenia, while the latter is that of canine granulocytic anaplasmosis. This work involved the detection and identification of Anaplasma species in blood samples from dogs in Colombia, using molecular techniques. Between December 2008 and April 2009, blood samples were drawn from the cephalic vein of 91 dogs in the central-western region of Colombia (cities of Bogota, Villavicencio and Bucaramanga) and stored in tubes containing EDTA. These samples were used in 16S rRNA-Anaplasma spp. nPCR and the preparation of blood smears. One (1.1%) of the 91 sampled dogs showed inclusions suggestive of Anaplasmataceae agents in the cytoplasm of platelets. Based on PCR followed by sequencing and phylogenetic analysis, A. platys and Anaplasma sp. closed related to A. phagocytophilum were detected in two and one dog, respectively. Interestingly, all the samples were negative for specific msp-2-A. phagocytophilum real-time qPCR, suggesting the circulation of an Anaplasma species phylogenetically related to A. phagocytophilum in dogs in the aforementioned region. Hence, Anaplasma spp. circulates among dogs in Colombia, albeit with low frequency. To the best of authors' knowledge, this is the first molecular detection of Anaplasma spp. in dogs in Colombia.Keywords: Dogs, Anaplasma spp., Anaplasma platys, Colombia. ResumoAnaplasma platys e A. phagocytophilum são patógenos transmitidos por carrapatos que parasitam plaquetas e neutrófilos, respectivamente, de humanos e animais. Enquanto o primeiro é agente etiológico da trombocitopenia cíclica canina, o último é o agente responsável pela anaplasmose granulocítica canina. O presente trabalho objetivou detectar e caracterizar, utiizando técnicas moleculares, a presença de espécies de Anaplasma em amostras de sague de cães na Colômbia. Entre Dezembro de 2008 e Abril de 2009, amostras de sangue colhidas em tubos contendo EDTA da veia cefálica de 91 cães da região centro-oeste da Colômbia (cidades de Bogotá, Villavicencio e Bucaramanga). As amostras de sangue foram utilizadas em reações de nPCR para 16S rRNA-Anaplasma spp. e confecção de esfregaços sanguíneos. Um (1,1%) dos 91 cães amostrados mostraram inclusões sugestivas de agentes Anaplasmataceae no citoplasma de plaquetas. Baseada na PCR seguida de sequenciamento e análise filogenética, A. platys and Anaplasma sp. relacionado a A. phagocytophilum foram detectados em dois e um cão, respectivamente. Interessantemente, todas as amostras mostraram-se negativas na PCR em tempo real quantitativa específica para msp-2-A. phagocytophilum, sugerindo a circulação de uma espécie de Anaplasma sp. filogeneticamente relacionado ao A. phagocytophilum em cães na região estudada. Embora em uma baixa freqüência, Anaplasma spp. circula entre cães na Colômbia. O presente trabalho representa a primeira detecção de Anaplasma spp. em cães na Colô...

Infección por Ehrlichia canis y Anaplasma sp. en caninos atendidos en clínicas veterinarias en Barranquilla, Colombia

“…Serological tests are only useful in the subclinical and/or chronic phase of the infection, which is unknown in this study; while the blood smear is useful in the acute phase, often asymptomatic, the low sensitivity in both natural (23) and experimental infections (24), and the low bacteraemia, make microscopic identification difficult (25) (23). Although the prevalence is low, the use of a microscopic technique that has been shown to be less sensitive, yields valuable data, taking into account that the prevalence reports in the country for commercial serological tests, Barranquilla 53.2% canine with A. platys, (15) and 40% canines with A. phagocytophilum (9), Cartagena 51% and Medellin 11% canines with A. phagocytophilum (9), could be limited to the low specificity or cross reactivity of some kits between Several species of Anaplasma and Ehrlichia (26).…”

“…Also, in dogs that presented co-infection, even when it was low, one dog developed pancytopenia, and reason why, it could not be ruled out the possibility of occurring in the other dogs, more complex hematological and clinical alterations as has been reported in other studies (14,23).…”

“…La prevalencia de Anaplasma sp. 11%, fue mayor comparada a lo reportado en Bucaramanga 1.1% caninos (23). A pesar de que la prevalencia es baja, por haberse utilizado una técnica microscópica, arroja datos valiosos, teniendo en cuenta los reportes de prevalencia por pruebas serológicas comerciales, Barranquilla 53.2% caninos con A. platys, (15) y 40% caninos con A. phagocytophilum (9), en Cartagena 51% y Medellín 11% caninos con A. phagocytophilum (9), podrían estar limitados a la baja especificidad o reactividad cruzada de algunos kits entre varias especies de Anaplasma y Ehrlichia (26).…”

Farmacocinética y efectos adversos de la doxiciclina en el tratamiento de la Ehrlichiosis: fundamentos teóricos para ensayos clínicos en caninos

“…CME is an infectious disease caused by the Ehrlichia canis bacterium, which is considered to be zoonotic (16,17) and emergent, transmitted mainly by the dog tick (Riphicephalus sanguineus) (18) phagocytophilum) en ejemplares caninos y de igual manera su detección en los vectores conocidos (19)(20)(21).…”

PCR detection of Anaplasma phagocytophilum in stray dogs in Batman, Turkey

“…In recent years, PCR method, which has a higher specificity and sensitivity than microscopic and serological diagnosis methods, has become a reason for preference in the diagnosis of anaplasmosis in dogs. By means of this technique, the infection can be detected even in the animals having very low parasitemia and thus reservoir animals can be determined (Massung and Slater 2003;Santos et al 2011;Ebani et al 2013;Vargas-Hernandez et al 2016;Huber et al 2017). In Turkey, the number of studies on the Anaplasma species in dogs is limited.…”