There is a great diversity of protein samples types and origins, therefore the optimal procedure for each sample type must be determined empirically. In order to obtain a reproducible and complete sample presentation which view as many proteins as possible on the desired 2DE gel, it is critical to perform additional sample preparation steps to improve the quality of the final results, yet without selectively losing the proteins. To address this, we developed a general method that is suitable for diverse sample types based on phenolchloroform extraction method (represented by TRI reagent). This method was found to yield good results when used to analyze human breast cancer cell line (MCF-7), Vibrio cholerae, Cryptocaryon irritans cyst and liver abscess fat tissue. These types represent cell line, bacteria, parasite cyst and pus respectively. For each type of samples, several attempts were made to methodically compare protein isolation methods using TRI-reagent Kit, EasyBlue Kit, PRO-PREPTM Protein Extraction Solution and lysis buffer. The most useful protocol allows the extraction and separation of a wide diversity of protein samples that is reproducible among repeated experiments. Our results demonstrated that the modified TRI-reagent Kit had the highest protein yield as well as the greatest number of total proteins spots count for all type of samples. Distinctive differences in spot patterns were also observed in the 2DE gel of different extraction methods used for each type of sample.

The aqueous extracts of thirty-four (34) tropical plants were tested in vitro for potential antimarine leech (Zeylanicobdella arugamensis) activity. The anti-leech activity was determined by exposing 8 adult leeches (Z. arugamensis) (9.3 ± 1.5 mm, aged 15 days) to 20 μl of plant extract (0.5 g/ml) for 5 min in a 24-well plate. After 5 min of exposure, the leeches were rinsed and transferred into seawater, to enable them to revive from the effect of the extract. Leech movements were observed from time to time and the numbers of paralyzed or dead leeches were recorded at 5, 20, 30 and 240 min. The efficacy of the plant extract in killing the adult Z. arugamensis during the 5 min exposure is reflected on the anti-leech property of the extract. The anti-leech property of positive plant extracts was also determined at different exposure time (1, 3, 5 min) and dilutions (1/2 (0.25 g/ml), 1/5 (0.1 g/ml) and 1/10 (0.05 g/ml). The extracts of 4 plants (Melastoma malabathricum, Piper betle, Tetracera indica and Etlingera coccinea) demonstrated anti-leech activity. The effects of M. malabathricum, P. betle and E. coccinea extracts on the leeches were very rapid causing death as early as a few seconds upon exposure. However, all four positive plant extracts were found not effective in killing the leeches at 1/10 dilution (0.05 g/ml). A Scanning Electron Microscopy examination on leeches exposed to the positive plant extracts exhibited effects such as protruding proboscis and shrunken or swollen bodies.

Renocila richardsonae Williams & Bunkley-Williams, 1992 is recorded for the first time from Sarawak coastal waters. During a survey of marine fishes at depth of 200 metres in South China Sea, off the coast of Sarawak from 3° to 5°N and 110° to 111°E, five specimens of Renocila richardsonae were collected from Japanese goatfish, Upeneus japonicus (Houttuyn, 1782) (Mullidae) with average prevalence of 17% and mean intensity of one isopod per fish from examination of 64 fish. The isopod was observed to cause tissue damage on the site of attachment to the host.