Amoebicidal activity of cationic carbosilane dendrons derived with 4-phenylbutyric acid against Acanthamoeba griffini and Acanthamoeba polyphaga trophozoites and cysts.

Amoebae from the genus Acanthamoeba are important pathogens responsible for severe illnesses in humans such as Acanthamoeba keratitis and granulomatous amoebic encephalitis. In the last few decades, AK diagnoses have steadily increased. Most patients suffering from AK were contact lens users and the infection was related to poor hygiene. However, therapy is not yet well established, and treatments may last for several months due to resistance. Moreover, these treatments have been described to generate cytotoxicity. Therefore, there is an urgent need to develop new therapeutic strategies against AK. In this study, the amoebicidal activity of different generation cationic carbosilane dendrons derived with 4-phenylbutyric acid was demonstrated against Acanthamoeba polyphaga and Acanthamoeba griffini trophozoites and cysts. In addition, the combination of chlorhexidine digluconate and the most effective dendron (ArCO2G2(SNMe3I)4) showed an in vitro effect against Acanthamoeba trophozoites and cysts, reducing the minimal trophozoite amoebicidal concentration as well as concentrations with cysticidal activity.

Molecular epidemiology of parasitic protozoa and Ehrlichia canis in wildlife in Madrid (central Spain).

Wildlife species are involved in the transmission of diverse pathogens. This study aimed to monitor raccoons (Procyon lotor), American minks (Neovison vison), and red foxes (Vulpes vulpes) as potential reservoirs in central Spain. Specifically, 200 spleen and fecal samples (from 194 raccoons, 3 minks, and 3 foxes) were analyzed molecularly by PCR/qPCR and sequencing for the presence of piroplasmids, Hepatozoon spp., Toxoplasma gondii, and Ehrlichia canis infections in the Community of Madrid (Spain). Biological samples were obtained in the years 2014, 2015, and 2016. No pathogen DNA was found in fecal samples. In contrast, analysis of raccoon spleen samples revealed that Toxoplasma was the most prevalent pathogen (prevalence 3.6 ± 2.6%), followed by Hepatozoon canis and E. canis (each with a prevalence of 2.57 ± 2.2%). Hepatozoon canis was also diagnosed in all three of the analyzed foxes. Analysis of yearly prevalence showed that tick-borne pathogens were less frequent in raccoon in 2015, a dry and warm year compared both to 2014 and 2016. These data suggest that fecal PCR assays are unsuitable for detection of DNA of non-erythrocytic pathogens. Furthermore, they demonstrate that the raccoon (an invasive species often living in proximity to domestic areas) and the red fox are putative reservoirs for pathogenic organisms in the Community of Madrid.

A survey for gregarines (Protozoa: Apicomplexa) in arthropods in Spain.

Gregarines thrive in the digestive tract of arthropods and may be deleterious to their hosts, especially when present in high densities. The impact of parasites on these invertebrates may affect both the ecosystem equilibrium and human economic activities. However, information available on gregarines in Spain is limited. Therefore, a microscopic study on prevalence of gregarine infection in 560 insects and crustaceans was undertaken in Madrid and Tarragona.Gregarina ormierei (78 % prevalence), Stylocephalus gigas (56 %), Oocephalus hispanus (13 %) and Actinocephalus permagnus (only one infected out of six beetles examined) were found in coleopteran hosts. Gregarina ovata and G. chelidurellae showed moderate frequency of infection (35 %) in dermapterans. An undescribed Gregarina sp. (76 % prevalence) was observed for the first time in freshwater decapod crustaceans. Interestingly, G. ormierei showed a noticeable phenotypic dimorphism, which justifies its redescription based on modern taxonomic criteria. Sequences of the 18S rRNA gene could be obtained only in the presence of highly prevalent gregarines. G. ormierei and Gregarina sp. were related (85 and 94 % identity by BLASTN, respectively) to G. basiconstrictonea and G. cloptoni, respectively, whereas S. gigas was closely related to both Xiphocephalus ellisi and S. giganteus (>97 % identity). Phylogenetic trees based on ribosomal sequences unequivocally grouped these new isolates either with the Gregarinidae (G. ormierei and Gregarina sp.) or the Stylocephalidae (S. gigas).