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The 1100-bp amplicons were purified and sequenced by a commercial laboratory (HyLabs, Rehovot, Israel). The resulting gyrB sequences were then aligned using the ClustalW algorithm to a series of 21 known sequences from 17 different previously characterized Aeromonas species using mega software version 4 (http://www.megasoftware.net/). A radial phylogenetic tree was constructed with the neighbour-joining method, using sequence data for reference strains of Aeromonas spp. derived from two studies [12,13] in which the same primers were used. Fisher��s exact test was used to compare treated with control leeches. Approximately 80% of the leeches engorged fully when fed artificially either on human blood or on the arginine/NaCl solution with and without Carnitine palmitoyltransferase II antibiotics. In 94.4% of the 106 unfed leeches treated with 70% ethyl alcohol no bacteria were grown from exterior surface samples. However, this treatment caused the death of the leeches as expected. Aeromonas spp. were detected in 57 out of 80 control leeches fed only with arginine/NaCl solution (71.3%), but in none of the 56 leeches treated with ciprofloxacin (p PD98059 clinical trial in the crop, in 35 animals (61.4%); they were confined to the crop in nine animals (15.8%), whereas in 13 (22.8%) they were present both in the crop and intestine. A blood-meal, through artificial Imatinib in vitro feeding, was offered to 56 leeches treated with antibiotics. Thirty-four of them took a blood meal 1�C4?weeks after treatment, whereas 22 leeches refused to feed, all of them in the first 2?weeks. All 56 leeches were dissected and examined for Aeromonas spp. All were negative (Table?1). Aeromonads were also not detected in the water in which the leeches were kept during the experimental period. Ten antibiotic-treated leeches were maintained individually in sterilized chlorine-free tap water containing ciprofloxacin at a concentration of 20?mg/L. Three died after 2?months, three after 3?months and four of them survived up to 4?months. Aeromonads were not detected in any of the 15 arginine/NaCl samples from feeding chambers that were used to feed antibiotic-treated leeches, whereas other environmental microorganisms were found in 16% (9 out of 56) of these leeches. Environmental bacteria such as Ochrobactrum spp. and some filamentous fungi were also isolated from 26.5% of treated leeches that had taken a blood meal 1�C4?weeks after treatment (data not shown). All 106 water samples in which leeches were kept before treatment were contaminated with Aeromonas spp. The dominant species was identified by the API 20NE kit as A. sobria. Two different phenotypes within this taxon were studied further: isolates designated 1 and 3. The API 20 NE profile for isolate 1 was 7176755 (99.2%. T 1.0).