Transcriptomic Variation of Amphiprion Percula (Lacepède, 1802) in Response to Infection with Cryptocaryon Irritans Brown, 1951.

Cryptocaryon irritans (Brown 1951) frequently infect the Pomacentridae fishes causing severe economic losses. However, the anti-C. irritans' molecular mechanism in these fishes remains largely unknown. To address this issue, we conducted RNA-Seq for C. irrtians-infected gills of the clownfish Amphiprion percula (Lacepède 1802) at the early (day 1) and late (day 3) stages of infection. A total of 1655 differentially expressed genes (DEGs) were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of DEGs showed a vast genetic variation related to the following aspects: ECM-receptor interaction, P13K-Akt signalling, cytokine-cytokine receptor interaction, and endocytosis. During the early phase of infection, key genes involved in ATP production, energy homeostasis, and stress control were abruptly increased. In the late phase, however, acute response molecules of the peripheral nervous system (synaptic transmission and local immunity), metabolic system triggering glycogen synthesis, energy maintenance, and osmoregulation were found to be critical. The highest number of upregulated genes (URGs) recovered during the early phase was included under the 'biological process' category, which primarily functions as response to stimuli, signalling, and biological regulation. In the late phase, most of the URGs were related to gene regulation and immune system processes under 'molecular function' category. The immune-related URGs of early infection include major histocompatibility complex (MHC) class-II molecules apparently triggering CD4+ T-cell-activated Th responses, and that of late infection include MHC class-1 molecules for the possible culmination of CD8+ T-cell triggered cytotoxicity. The high level of genic single nucleotide polymorphisms (SNPs) identified during the late phase of infection is likely to influence their susceptibility to secondary infection. In summary, the identified DEGs and their related metabolic and immune-related pathways and the SNPs may provide new insights into coordinating the immunological events and improving resistance in Pomacentridae fishes against C. irritans.

Immune responses of Cyprinus carpio induced by protein extracts of Lernaea cyprinacea Linnaeus, 1758.

Lernaea cyprinacea Linnaeus, 1758 is an ectoparasite showing widespread infections in tropical aquaculture, and the present study aimed to determine the specific immune responses against this parasite. For the experiment, whole parasite extracts were injected intraperitoneally into Cyprinus carpio Linnaeus, 1758, and samples of epidermal mucus and blood were drawn at 0, 1, 7 and 14-days post-injection (DPI). The results revealed high levels of protein, protease and lysozyme activities in the experimental fish which were injected with L. cyprinacea protein extract. In the epidermal mucus, the total protein concentration of the control fish was 460 µg/mL, and the level raised significantly to 800 µg/mL in the experimental fish. The lysozyme activity increased from 741.5 u/mL to a peak level of 1448.5 u/mL at 7DPI. The protease activity was also found elated gradually from 2.91 u/µL to 4.49 u/µL at 1 to 14 DPI. In the serum samples, the protein concentration remained steady throughout the experiment period. However, all the experimental fish displayed statistically high levels of lysozyme and protease activity, from 890 u/mL to 1220 u/mL, and 6.10 u/µL to 11.88 u/µL, respectively. In the whole blood samples, the haemoglobin content and the red blood cells (RBC) count did not show any significant change in any of the experimental groups. But, the percentage of lymphocytes showed a marginal increase from 0.47 to 0.6 in the experimental groups. Overall, the immune responses induced by L. cyprinacea protein extracts depicts a pattern of specific responses, in which the local humoral responses dominate the systemic humoral/cellular response. The results further revealed the possibility of futuristic approaches to control freshwater ectoparasites.