METHODS: Fish were collected off Mazatlán Port (23° 12' N, 106° 26' W), in the State of Sinaloa, Mexico (southeastern Gulf of California). The copepods were morphologically analyzed by light microscopy. Sequences of the COI mtDNA gene were generated for the first time for this species. These sequences were compared to COI sequences from six species of Lernaeenicus available in GenBank.
RESULTS: The specimens of the present study exhibited a cephalosome without apparent lateral processes, which were originally described for L. longiventris. No remarkable differences were observed with previous descriptions regarding appendages and body proportions. The phylogenetic tree based on COI sequences showed that L. longiventris was closer to L. radiatus although with low bootstrap values support in ML tree, both species formed a sister clade of L. sprattae.
CONCLUSIONS: Lernaeenicus longiventris is the unique species of the genus in the Mexican Pacific and the Gulf of California, and also the unique species of Lernaeenicus infecting C. caninus. Molecular data of L. longiventris from host and locality type are required to avoid misidentification of this species.
METHODS: In this study, A. triangularis amoebas were treated with a sublethal dose of curcumin under nutrient starvation and nutrient-rich condition and the surviving amoebas was investigated. Cysts formation and vacuolization were examined by microscopy and transcriptional expression of autophagy-related genes and other encystation-related genes were evaluated by real-time PCR.
RESULTS: A. triangularis cysts were formed under nutrient starvation. However, in the presence of the autophagy inhibitor, 3-methyladenine (3-MA), the percentage of cysts was significantly reduced. Interestingly, in the presence of curcumin, most of the parasites remained in the trophozoite stage in both the starvation and nutrient-rich condition. In vacuolization analysis, the percentage of amoebas with enlarged vacuole was increased upon starvation. However, the percentage was significantly declined in the presence of curcumin and 3-MA. Molecular analysis of A. triangularis autophagy-related (ATG) genes showed that the mRNA expression of the ATG genes, ATG3, ATG8b, ATG12, ATG16, under the starvation with curcumin was at a basal level along the treatment. The results were similar to those of the curcumin-treated amoebas under a nutrient-rich condition, except AcATG16 which increased later. On the other hand, mRNA expression of encystation-related genes, cellulose synthase and serine proteinase, remained unchanged during the first 18 h, but significantly increased at 24 h post treatment.
CONCLUSION: Curcumin inhibits cyst formation in surviving trophozoites, which may result from its effect on mRNA expression of key Acanthamoeba ATG-related genes. However, further investigation into the mechanism of curcumin in A. triangularis trophozoites arrest and its association with autophagy or other encystation-related pathways is needed to support the future use of curcumin.