Molecular surveillance of tick-borne diseases affecting horses in Poland-Own observations.

The purpose of the study was to carry out the molecular surveillance of piroplasmosis, granulocytic anaplasmosis and lyme borreliosis in horses which originated from Poland and exhibited symptoms raising the suspicion of the aforementioned disease units. The presence of Theileria equi genetic material was detected in 37 out of 512 examined horses (7.2%), and Anaplasma phagocytophilum in 9 (1.8%). The DNA of Borrelia burgdorferi was found in 11 out of 204 examined horses (5.4%). The above-cited results indicate that the problem of tick-borne diseases affecting horses in Poland is not as significant as in other parts of Europe, however they have to be considered in differential diagnosis of the diseases with lethargy, fever, anaemia and thrombocytopenia.

Naphthylisoquinoline alkaloids and their synthetic analogs as potent novel inhibitors against Babesia canis in vitro.

Babesia canis is the predominant and clinically relevant canine Babesia species in Europe. Transmitted by vector ticks, the parasite enters red blood cells and induces a severe, potentially fatal hemolytic anemia. Here, we report on the antibabesial activities of three extracts of the West African tropical plant species Triphyophyllum peltatum (Dioncophyllaceae) and Ancistrocladus abbreviatus (Ancistrocladaceae) and of 13 genuine naphthylisoquinoline alkaloids isolated thereof. Two of the extracts and eight of the alkaloids were found to display strong activities against Babesia canis in vitro. Among the most potent compounds were the C,C-coupled dioncophyllines A (1a) and C (2) and the N,C-linked alkaloids ancistrocladium A (3) and B (4), with half-maximum inhibition concentration (IC50) values of 0.48 µM for 1a, 0.85 µM for 2, 1.90 µM for 3, and 1.23 µM for 4. Structure-activity relationship (SAR) studies on a small library of related genuine analogs and non-natural synthetic derivatives of 1a and 2 revealed the likewise naturally occurring alkaloid N-methyl-7-epi-dioncophylline A (6b) to be the most potent (IC50, 0.14 µM) among the investigated compounds. Although none of the tested naphthylisoquinolines showed 100 % inhibition of parasite infection - as displayed by imidocarb dipropionate (IC50, 0.07 µM), which was used as a positive control - the antibabesial potential of the dioncophyllines A (1a) and C (2) and related compounds such as 6b, its atropo-diastereomer 6a (IC50, 1.45 µM), and 8-O-(p-nitrobenzyl)dioncophylline A (14) (IC50, 0.82 µM) is to be considered as high. The SAR results showed that N-methylation and axial chirality exert a strong impact on the antibabasial activities of the naphthylisoquinolines presented here, whereas dimerization, as in jozimine A2 (5) (IC50, 140 µM), leads to a significant decrease of activity against B. canis. Alkaloids displaying good to high activities against B. canis like the dioncophyllines 1a, 2, 6a, and 6b were found to cause only a small degree of hemolysis (< 0.7 %), whereas compounds with moderate to weak antibabesial activities such as 6-O-methyl-4'-O-demethylancistrocladine (15a) (IC50, 14.0 µM) and its atropo-diastereomer 6-O-methyl-4'-O-demethylhamatine (15b) (IC50, 830 µM) caused a high degree of hemolysis (7.3 % for 15a and 11.2 % for 15b). In this respect, the most effective anti-Babesia naphthylisoquinolines are also the safest ones.