Endotoxins (chemically known as Lipopolysaccharide) from gram-negative microorganisms initiates clot formation in blood when it is accidentally encountered by horseshoe crab blood stream. This property was extensively studied by various researchers as a result Limulus Amebocyte Lysate (LAL) test was established. The LAL tests in general, 3 to 300 times more sensitive than the United States Pharmacopeial (USP) rabbit pyrogen test method. It is apparent that major differences among the LAL preparations lie in the area of sensitivity. Differences, up to 100-fold, exist in the sensitivity of the various LAL preparations to the same endotoxin. Based on the above perspective, a portable Kit (Endo sensor) was developed to detect the presence of bacterial endotoxin in liquid biological samples using Tachypleus Amebocyte Lysate (TAL) as a source. Sensitivity of the Kit was determined using various concentrations of prepared endotoxin standards and pyrogen free water samples. It was observed that Endo sensor could detect up to nano gram level of endotoxin in liquid biological samples which could be expressed in (EU/ml) and the labeled sensitivity of the lysated product was 0.125 EU/ml. The gel clotting principle method was utilized for the detection of bacterial endotoxin in liquid biological samples.
Increasing utilization of living animals in ecological and biomedical research has drawn serious concerns in terms of animal welfare and ethical practices in animal handling. Significant attention has been given to animals of higher taxonomical hierarchy especially vertebrates such as fishes, rodents, reptiles and mammals, while ethical framework on invertebrate handling and welfare is less addressed (except for cephalopods). The definition of ‘Animal’ itself by any international consortia or Animal Research Act (ARA) does not include invertebrates as an animal entity. This is due to the lack of standard ethical framework to understand the pain and other physiological stress experienced by the invertebrate test animal. One such example would be the living fossil ‘horseshoe crab’ which is extensively bled to obtain its blue blood that is used for endotoxin quantification in biological samples. The biomedical bleeding itself leads to 15-30% post bleeding mortality of crabs, while pain and stress caused by the bleeding practice is not studied. Hence, this paper discusses the technicality of establishing standard framework for invertebrate handling. The paper also highlights the shari’ah (Islamic law) principles on scientific experimentations on animal subjects, particularly the norms related to the adoption of invertebrates in environmental and biomedical practice. Comprehensive review of ethical regulations in animal experiments, especially invertebrates, would be beneficial for revising and improving existing animal ethical practices.
Microorganisms are the major cause of spoilage of most seafood products. Fishes are more perishable than other protein foods and thus more prone to bacterial contamination. Based on above perspectives, a bacterial invasion in commercially important fresh and spoiled marine (Lates calcarifer, Lutjanus sanguineus) and freshwater fish (Pangasius pangasius) were analyzed using API 20E kit. Out of 25 isolates obtained from fresh water fish, only 6 isolates were characterized as Gram-positive bacteria and the rest were Gram negative strains (19 isolates). The most dominant genera were Vibrio, Enterobacter, Serratia, and Aeromonas. All these bacteria were found in both fresh fish and spoiled fish sample while Erwinia spp. and Kluyvera spp. were identified only in fresh fish samples. Out of four (4) strains of Staphylococcus spp., S. xylosus was detected exclusively from spoiled fish. The higher number of bacterial micro flora in the spoiled fish gut indirectly indicated increased microbial degradation in the fish gut during spoilage process. Notably, almost all the isolates were lactose degraders, positive oxidizers and carbohydrate fermenters. Vibrio fluvialis, Proteus mirabilis, Proteus vulgaris, Brucella sp. and Ochrabactrum anthropi were the human pathogenic bacteria found in marine fish Lates calcarifer (Sea perch). While Vibrio fluvialis, Proteus mirabilis and Proteus vulgaris were detected in Lutjanus sanguineus (Red snapper). The study portrays that the existing postharvest handling techniques could be a vital factor for degrading hygienic conditions of fish in local fish markets. Nevertheless, a long term monitoring is an urgently needed for sustaining the quality flesh of fish towards the betterment of the consumer’s health.
The data available in this repository were gathered from Balok, the only most productive spawning site for horseshoe crabs Tachypleus gigas and Carcinoscorpius rotundicauda in East Coast of Peninsular Malaysia. The mangrove horseshoe crab, C. rotundicauda population and spawning data are available in the first table. The horseshoe crabs were retrieved from Balok River using 11.43 cm mesh size gill nets installed at the river mouth, the confluence and last meander. The arthropods were inspected for damage, abnormality and growth before their release into Balok River, particularly at the site of capture. Sediment samples were retrieved at their spawning grounds to ascertain sediment composition and size classifications which were also processed using Logarithmic Method of Moments. Water parameters like temperature, pH and salinity were also investigated during year 2016. All these information are compiled into the second table and arranged according to the period of data availability. The horseshoe crab catch data of years 2012, 2013, 2014, 2015 and 2016 were made available by artisanal fisher and compiled in the third and fourth table for inter-species comparison.
Carcinoscorpius rotundicauda and Tachypleus gigas may co-exist and share common spawning grounds elsewhere but at Balok (East Coast of Peninsular Malaysia), C. rotundicauda is an understudied species. Neglected as research candidate because of inaccessible spawning grounds, smaller size and less commercial value than T. gigas and also, difficulty to attain from the wild has made C. rotundicauda population status remaining unidentified at Balok. This standpoint drove the present attempt because anthropic activities like structure placement and mining are point-source for runoffs that load sediments into Balok River. While erosion-accretion events have altered Balok River width, the shore sediments in Balok Beach were transitioned between medium-fine and fine sand between years 2012 and 2016. Eventually by year 2016, the C. rotundicauda were depositing 5117 eggs in 91 nests from 200 to 1000 m range along this corridor facing South China Sea. From this yield, C. rotundicauda released 2880 eggs in 56 nests during the Southwest monsoon, 1254 eggs in 19 nests during the Northeast monsoon and 983 eggs in 16 nests during the Inter-monsoon seasons. Though female C. rotundicauda opted to lay their eggs in shallow burrows at lower shorelines, the absence of erosion and substantial silt and clay (>20%) deposition facilitates C. rotundicauda embryogenesis with brief periods of temperature and salinity shocks during day-time falling tides. This encourages C. rotundicauda to emerge with increasing abundance and carry out bi-monthly spawning at Balok Beach. In short, shore restoration initiatives like systematic boat docking, proper disposal of nets and waste and, periodic fish-catching operations were effectively led by the Balok fisher citizen scientist. This successful community joint-cooperation proves that citizen-led caretaking of degraded beaches offers marine life protection and are practical for coastal area management especially at areas where other oviparous animals such as turtles and crocodiles are harboured.