Positive amplicons were gel-purified (NucleoSpin Extract, Macherey-Nagel Inc., Bethlehem, PA, USA) and directly sequenced. The obtained sequences were selleck screening library aligned using ClustalW with available sequences for the E. granulosus genotypes. Table 1 summarizes genes, primers, sequences used for comparisons and corresponding accession numbers in GenBank.Table 1Genes, primer sequences, and accession numbers used for genetic comparisons.3. Results3.1. Macro- and Microscopical ExaminationMorphological analysis of the cysts removed from the liver (one from each animal) revealed that they were subspherical in shape, 3�C5 �� 4�C6cm, unilocular, fluid-filled, and containing a different number of protoscolices (one, two, three, two) (Table 2).

The rostella consisted of two rows of alternating large and small hooks (34�C37 in number); large hooks were 25�C27��m in total length and 12-13��m in blade length, whereas small ones were 19�C22��m in total length and 9�C12��m in blade length. Figure 1 shows some large and small hooks (a) isolated from the rostellum and in preparation for counts and measurements of the parameters considered as valid for identifying E. granulosus strains (b).Figure 1(a) Some large and small hooks isolated from the rostellum. (b) Diagram of measurements made on protoscolex rostrellar hooks (from [5]).Table 2Main features of the hydatid cysts found in goat liver.3.2. Molecular AnalysesMolecular identification proved the strains involved in the infection to be highly identical to the G3 buffalo strain. In fact, the analysis of the variable sites of the cox1 sequences obtained for the four samples indicates 99.

5% identity to G3 and 99.3% and 99.1% to G2 and G1, respectively (Table 3). The genetic identity to the other recognized species was lower: 93.4% to E. equinus, 92.9�C92.7% to E. canadensis, and 92.7% to E. ortleppi. Sequencing of the nad1 and 12S rRNA supports these results. Blast identity search evidenced higher identity to the G3 (99.78% and 99.64%, resp.) than to G1 strain (99.30% and 98.93%, resp.).Table 3Alignment of the variable sites Cilengitide in the partial cox1 mitochondrial gene of the isolates of E. granulosus sensu lato evidenced in the goats (goat 1�C4) examined in Central Italy with available sequences for other genotypes deposited in GenBank (E.g1�C7). …4. DiscussionTo the best of our knowledge, strains to date reported in goats are the sheep strain G1 (widely distributed), the cattle G5 and the pig G7 strains (present also in Europe), and the camel strain G6, probably absent from Europe [6, 8�C10].