Parasite species often show a heterogeneous, highly dispersed pattern of infestation within hosts. Varieties of factors including morphological, physiological, immunological and nutritional characteristics affect the infestation level of a specific parasite in homogenous pray. Limited attempts, however, have been made to explore such underlying drivers of infestation pattern. Here, three stages of Labeo rohita (fingerling, juvenile and pre-adult) were challenged with ectoparasite, Argulus siamensis in same aquaria. The parasite load on individuals was determined at 5-day interval for 1 month. The load was found to be highest in pre-adult stage followed by juveniles and fingerlings. On day 20 post infection, the load of parasite on pre-adult fish was high along with detectable skin damages. Skin tissues were collected for immune gene expression analysis and histopathology. Histological studies showed increased melanization in the dermis and mild inflammatory cellular reactions in pre-adult fish whereas, massive subcutaneous myositis with engorged blood vessels were observed in fingerlings. The expression levels of various inflammation and innate immune-related genes viz., interleukin (IL)-8, IL-10, IL-11, IL-15, natural killer enhancing factor, toll-like receptor 4, apolipoprotein A-I and immunoglobulin Z were significantly high in skin samples of infected fingerlings as compared to other two growth stages or controls of each stage. On the other hand, the expression of immunoglobulin M was down-regulated in all infected samples as compared to their respective controls. The results thus depict that local immuno-inflammatory response plays a significant role in determining susceptibility of host in intra-specific group, and has important implications for ecology and aquaculture.
Argulus spp. are economically important fish ectoparasites. The development of antiparasitic drugs is thus important and real time PCR is an indispensable tool in drug development. The analytical potential of RT-PCR depends upon accurate normalisation by the use of stable reference genes. Here, we identified stable reference genes of Argulus siamensis for validation of efficacy of drugs and drug targets. Seven candidate genes were evaluated by evaluating their expression under different states of Argulus using the RefFinder tool. The four algorithms together generated a comprehensive ranking with elongation factor-1 alpha (EF-1α) being the most stable and 18S ribosomal protein (18S) the least stable gene. Taking EF-1α and 18S genes as references, the effectiveness of six anti-parasitic compounds against Argulus was evaluated by studying their effect on the expression pattern of few ion channel genes; this was to understand their mode of action, besides validating the reference genes. EF-1α was found to be the most stable gene in the validation. Collectively, this study is the first report to validate the optimal reference genes of A. siamensis for normalisation, and the potential of the ion channel genes for evaluating effective drug targets in parasite control.
T HE PRIMARY objective of this study is to present a database of indigenous knowledge on medicinal plants used to cure diseases among the local traditional people of Bhadrak district of Odisha, which could provide novel remedies for diseases. The study highlights the use of local flora (wild and cultivated plant species); explore; identify; ethnobotany and conservation of Bhadrak district. It also pays heed to the proper utilization of flora as a way of highlighting its ethnopharmacological importance. A field survey was conducted to collect the information about floral diversity in and around the study site. Data on the use of plants were collected with a semi-structured questionnaire and from the peer reviewed literatures. A total of 85 plant species belongs to 46 different families were identified and documented along with their botanical name. Among them Cucurbitaceae, Asteraceae, Euphorbiaceae, Solanaceae, Amaranthaceae, Fabaceae and Rutaceae were found to be dominated families. It is concluded that many people in the studied area still depend on medicinal plants for their treatment of different diseases like urinary tract infections, gastrointestinal tract infections, piles, tumors, asthma, diabetes, etc. This study needs to harness this traditional knowledge and preserve for the betterment of future mankind.
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