Unveiling antimicrobial activity of microalgae Chlorella sorokiniana (UKM2), Chlorella sp. (UKM8) and Scenedesmus sp. (UKM9).

Microalgae represent promising sources of bioactive compounds for pharmaceutical and industrial applications. The emergence of antibiotic resistant bacteria leads to the need to explore new cost-effective, safe, and potent bioactive compounds from the microalgae. This study aimed to investigate the potential of local microalgae for their antimicrobial properties and bioactive compounds. Three local microalgae namely Chlorella sorokiniana (UKM2), Chlorella sp. UKM8, and Scenedesmus sp. UKM9 biomass methanol extracts (ME) were prepared and tested against Gram-positive and Gram-negative bacteria. Chlorella sp. UKM8-ME showed the highest antibacterial activity. UKM8-ME minimum inhibitory concentrations were in the range of 0.312 to 6.25 mg/mL. Cytotoxicity evaluation using MTT assay showed that the microalgae methanolic extracts did not exhibit cytotoxicity against Vero-cells. The UKM8-ME was mainly containing 28 compounds from the Gas Chromatography-Mass Spectrometry (GC-MS) analysis. Major compounds of UKM8-ME included phenol (18.5%), hexadecanoic acid (18.25%), phytol (14.43%), 9,12-octadecadienoic acid (13.69%), and bicyclo[3.1.1]heptane (7.23%), which have been previously described to possess antimicrobial activity. Hence, Chlorella sp. (UKM8) methanol extracts showed promising antibacterial activity. More comprehensive studies are required to purify these antimicrobial compounds and develop our understanding on their mechanism in UKM8-ME to unleash their specific potential.

Localization of tetrodotoxin in the skin of a brackishwater puffer Tetraodon steindachneri on the basis of immunohistological study.

A monoclonal antibody-based immunoassay for localization of tetrodotoxin (TTX) in the skin of a brackishwater puffer Tetraodon steindachneri is described in this paper. TTX was recognized in the undifferentiated basal cells and succiform cells in the skin under light microscope. Malpighian cells of the skin did not exhibit any TTX antigen. Neither gland nor enclosed gland-like apparatus possessing TTX was apparent in the skin.

Chelonodon patoca, a highly toxic marine puffer in Japan.

Toxicity of a Japanese marine puffer Chelonodon patoca ("okinawafugu") was examined by mouse assay from 1996 to 1999. Frequency of the toxic specimens was found to be 100% with high toxicity scores. Among the tissues tested, toxicity in the skin ranged from 60 to 6,700 MU/g, in the ovary from 25 to 670 MU/g, in the testis from 45 to 550 MU/g, in the muscle from 2 to 390 MU/g, and in the liver from 5 to 380 MU/g. The liver, which is known as one of the most toxic organs in Japanese marine puffer in general, showed lower toxicity in the present study. Thus, the anatomical distribution of toxicity was unique in C. patoca, in comparison with that of other Japanese puffers. C. patoca toxin was characterized as tetrodotoxin (TTX), 4-epiTTX and anhydroTTX by HPLC.

Occurrence of tetrodotoxin in the skin of a rhacophoridid frog Polypedates sp. from Bangladesh.

Twenty-three specimens of a tree-frog Polypedates sp. were collected from two locations (Mymensingh and Barisal) of Bangladesh in 1999, and assayed for their toxicity scores and toxin principle. Among the tissues, only the skin of the Mymensingh specimens was found to be toxic in mouse test, with the toxicity scores of 31-923MU/g. The toxin isolated from the skin was analyzed by high-performance liquid chromatography, electrospray ionization-time of flight mass spectrometry and proton nuclear magnetic resonance, and characterized as tetrodotoxin, a toxin principle.

Delayed haemolytic activity by the freshwater puffer Tetraodon sp. toxin.

In order to elucidate the toxin composition of the freshwater puffer in Bangladesh, about 230 specimens of Tetraodon sp. were collected from 1997 to 1999 and extracted. After partitioning the toxins between an aqueous layer and a 1-butanol layer, the toxin in the aqueous layer was characterized as paralytic shellfish poison (PSP) (data not shown), while the toxin in the 1-butanol layer was identified as palytoxin (PTX) or PTX-like substance based on the delayed haemolytic activity which was inhibited by an anti-PTX antibody and ouabain (g-strophanthin). This is the first report on the occurrence of PTX or PTX-like substance(s) in puffer fish.

An epidemic survey on freshwater puffer poisoning in Bangladesh.

An epidemic investigation was carried out on freshwater puffer poisoning incidents in Bangladesh from April 1988 to May 1996. A lot of information on 10 poisoning cases involving 55 victims was collected through newspapers, interviewing the victims and their families, concerned hospital sources or questionnaires to them. Symptoms of the victims were partly similar to those caused by paralytic shellfish poison (PSP) or tetrodotoxin (TTX). Among them, however, muscle pain, discharge of black urine, and longer recovery time are clearly different. Further, serum creatine phosphokinase (CPK) values were found to be higher (230-450 and 298-430 IU/l) than normal values in two cases. From these different symptoms and high CPK values, it can be predicted/assumed that present freshwater puffer toxin is implicated in not only PSP, but also other toxin(s).