Heavy infection of Ichthyophthirius multifiliis in striped catfish (Pangasianodon hypophthalmus,Sauvage 1878) and its treatment trial by different therapeutic agents in a control environment

ABSTRACT

Striped catfish (Pangasianodon hypophthalmus) fingerlings were stocked (60 fishes/m³) in cemented square tanks for ornamental fishery purposes at the College Fish Farm in Mangalore. A total of 400 fishes with a mean weight (g) ranging from 3.24 ± 1.21 to 6.70 ± 1.13 and a mean length (cm) of 7.50 ± 0.94 to 9.50 ± 1.10 were examined. A severe outbreak of salt-like granule white spots was found on the body surface. Ichthyophthirius multifiliis, a ciliate pathogen, was identified as the causative agent by clinical signs, wet mount, and histopathological observations. Infected fishes were transferred and equally distributed to the 0.45 m³ glass aquaria and treated with three treatments: (T1) methylene blue + salt; (T2) raising temperature with salt; and (T3) formalin + malachite green. The best fingerling survival (55 ± 9.36%) was obtained by elevated water temperature with salt in T2.     

Polyclonal antibody‐based farmer‐friendly flow‐through test for the detection of acute hepatopancreatic necrosis disease in shrimp

Early mortality syndrome (EMS) or acute hepatopancreatic necrosis disease (AHPND) is currently the most significant disease of shrimp in farms of Vietnam, Thailand, Malaysia, China and Mexico, and there is a great risk that it may spread to other shrimp farming countries. Although, an array of sophisticated detection tools for AHPND available, there is a need for a sensitive, simple and rapid detection method. In this study, a simple, sensitive, rapid and polyclonal antibody‐based farmer‐friendly flow‐through assay (FTA) test has been developed for the detection of AHPND pathogen. The recombinant Photorhabdus insect‐related (Pir) A toxin‐like protein of AHPND pathogen was used to immunize rabbits at 21‐day interval observed for highest antibody titre after third booster by ELISA. The raised rabbit antiserum was purified by affinity chromatography and characterized by Western blot. The antiserum showed no cross‐reactivity with AHPND‐free Vibrio parahaemolyticus, V. anguillarum, White Spot Virus (WSV), Aeromonashydrophila and Aphanomycesinvadans. This polyclonal rabbit antiserum was used to develop a farmer‐friendly FTA test for the detection of AHPND pathogen. This simple FTA testis is more sensitive and could detect PirA^VP toxin up to 0.121 µg/ml, compared with 0.242 µg/ml by immunodot assay. Furthermore, FTA test requires only 8–10 min for completion, compared with 3 hr by immunodot thus found to be more sensitive, specific and cost‐effective. Collectively, sensitive FTA test would help shrimp farmers to take real‐time management decisions, especially emergency harvest and finally be a better hope for the prevention of AHPND.     

Hydrological impacts of afforestation — A review of research in India

We review the current status of experimental studies in India to understand the linkages between afforestation and hydrology. This discipline deals with the use and management of water, social awareness of environmental problems, and ecosystem limitations on provision of watershed services by river basins in the mountain regions of India. Our review begins with examination of experimental works in tropical countries and proceeds to discussion of initiatives of Indian research institutes and Government organizations towards establishing experimental watersheds to understand the impacts of land cover changes on hydrologic regimes in the Indian sub-continent. This is followed by the review of the experimental work carried out by various authors to assess the impact of land cover changes on major water balance components such as, runoff, groundwater, evapotranspiration and sediment yield. The spatial scales of these experiments have been limited to small watersheds or field plots. This paper also describes impacts of human interventions （such as planta- tions of exotic forest species to restore degraded landscapes） on the water balance components in diverse hydro-geo-environmental conditions in the Indian sub-continent. We focus on identifying the research areas which require immediate attention to develop tools to assist planners and policy makers in assessing and managing the water resources in these complex environments. The review is largely based on research results reported during the last 20 to 30 years.     

Development and standardization of a monoclonal antibody-based rapid flow-through immunoassay for the detection of Aphanomyces invadans in the field

A monoclonal antibody-based flow-through immunoassay (FTA) was developed using a nitrocellulose membrane placed on the top of adsorbent pads enclosed in a plastic cassette with a test zone at the center. The FTA could be completed within 10 min. Clear purple dots against a white background indicated the presence of Aphanomyces (A.) invadans. The FTA limit of detection was 7 µg/mL for A. invadans compared to 56 µg/mL for the immunodot. FTA and polymerase chain reaction (PCR) could detect A. invadans in fish tissue homogenates at a 10^-11 dilution compared to a 10^-8 dilution by immunodot. In fish suffering from natural cases of epizootic ulcerative syndrome (EUS) collected from Mangalore, India, FTA and PCR could detect A. invadans in 100% of the samples compared to 89.04% detected by immunodot. FTA reagents were stable and produced expected results for 4 months when stored at 4~8℃. This rapid test could serve as simple and cost-effective on-site screening tool to detect A. invadans in fish from EUS outbreak areas and in ports during the shipment of live or frozen fish.     

Monoclonal antibody-based immunodot for evaluating serum immunoglobulin levels in rohu (Labeo rohita)

Immune response in rohu treated with an immunomodulator is usually evaluated employing either non-specific immune parameters or traditional antibody-based tools. In the present study, a monoclonal antibody-based immunodot has been developed for evaluating antibody titre in rohu as a preliminary tool to ensure antibody response due to the effect of an immunomodulator, which can be used for routine field level analysis. The immunodot was sensitive enough to determine rohu immunoglobulin up to 15 μg/ml. Application of the immunodot for evaluating enhancement in immune response could be successfully demonstrated in probiotic fed rohu.     

Development of a simple and rapid monoclonal antibody-based flow through immunogold assay (FIA) for detection of <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

In this study, we developed a simple, low-cost, and rapid flow through immunogold assay (FIA) for detection of Aeromonas hydrophila in fish tissues and validated. The developed assay relied on colloidal gold conjugated highly specific monoclonal antibody (B8E3) against 20 kDa protein of A. hydrophila. The assay can be completed within 5 min including antigen (sample) preparation and exhibited no cross-reaction with other major aquatic pathogens such as Vibrio anguillarum, V. parahaemolyticus, V. harveyi, white spot syndrome virus (WSSV), and Aphanomyces invadans. A wider range of Aeromonas species were tested including A. caviae, A. dhakensis, A. sobria, A. veronii, A. culicicola, A. sharmana, and A. schubertii and no cross-reactivity could be observed. The developed technique FIA could detect 10³ CFU/ml of aeromonad cells. In addition, the FIA is as sensitive as the first-step polymerase chain reaction (PCR) and 1000 times sensitive than the immunodot blot method. The validation study revealed the accuracy of FIA by comparing with the standard first-step PCR. The proposed FIA is low-cost, easy-to-use, and compatible for field-level analysis of A. hydrophila in fish tissues. Our developed FIA could successfully detect A. hydrophila at early stage of the infection and could be easily availed by farmers, fish producers, non-technicians, and technicians in the aquaculture industry.