Porphyrin detection in denatured cnidarian tissue extracts

Porphyrin metabolic disruption from exposure to xenobiotic contaminants such as heavy metals, dioxins, and aromatic hydrocarbons can elicit overproduction of porphyrins. Measurement of porphyrin levels, when used in conjunction with other diagnostic assays, can help elucidate an organism’s physiological condition and provide evidence for exposure to certain toxicants. A sensitive microplate fluorometric assay has been optimized for detecting total porphyrin levels in detergent solubilized protein extracts from symbiotic, dinoflagellate containing cnidarian tissues. The denaturing buffer used in this modified assay contains a number of potentially interfering components (e.g., sodium dodecyl sulfate (SDS), dithiothreitol (DTT), protease inhibitors, and chlorophyll from the symbiotic zooxanthellae), which required examination and validation. Examination of buffer components were validated for use in this porphyrin assay; while the use of a specific spectrofluorometric filter (excitation 400 ± 15 nm; emission 600 ± 20 nm) minimized chlorophyll interference. The detection limit for this assay is 10 fmol of total porphyrin per μg of total soluble protein and linearity is maintained up to 5000 fmol. The ability to measure total porphyrins in a SDS protein extract now allows a single extract to be used in multiple assays. This is an advantage over classical methods, particularly when tissue samples are limiting, as is often the case with coral due to availability and collection permit restrictions.

Spatial and temporal variation in the diet of the California sea lion (Zalophus californianus) in the Gulf of California, Mexico

Between June 1995 and May 1996 seven rookeries in the Gulf of California were visited four times in order to collect scat samples for studying spatial and seasonal variability California sea lion prey. The rookeries studied were San Pedro Mártir, San Esteban, El Rasito, Los Machos, Los Cantiles, Isla Granito, and Isla Lobos. The 1273 scat samples collected yielded 4995 otoliths (95.3%) and 247 (4.7%) cephalopod beaks. Fish were found in 97.4% of scat samples collected, cephalopods in 11.2%, and crustaceans in 12.7%. We identified 92 prey taxa to the species level, 11 to genus level, and 10 to family level, of which the most important were Pacific cutlassfish (Trichiurus lepturus), Pacific sardine (Sardinops caeruleus), plainfin midshipman (Porichthys spp.), myctophid no. 1, northern anchovy (Engraulis mordax), Pacific mackerel (Scomber japonicus), anchoveta (Cetengraulis mysticetus), and jack mackerel (Trachurus symmetricus). Significant differences were found among rookeries in the occurrence of all main prey (P≤0.04), except for myctophid no. 1 (P>0.05). Temporally, significant differences were found in the occurrence of Pacific cutlassfish, Pacific sardine, plainfin midshipman, northern anchovy, and Pacific mackerel (P<0.05), but not in jack mackerel (χ 2=2.94, df=3, P=0.40), myctophid no. 1 (χ 2=1.67, df= 3, P=0.64), or lanternfishes (χ 2=2.08, df=3, P=0.56). Differences were observed in the diet and in trophic diversity among seasons and rookeries. More evident was the variation in diet in relation to availability of Pacific sardine.

Health and disease status of Thai pangas, Pangasius hypophthalmus cultured in rural ponds of Mymensingh, Bangladesh

Thai pangas, Pangasius hypophthalmus is one of the important aquaculture species in Bangladesh. Over the last few years spectacular development has been taking place in Thai pangas farming in Mymensingh district. Due to availability of easy breeding and culture techniques as well as quick return, more and more people are converting their rice fields into pangas farms overnight. The present study was carried out to examine health and disease status of Thai pangas mainly through clinical, histopathological and bacteriological techniques. In addition, for collecting primary data on disease and health status of Thai pangas and the resultant socioeconomic impacts on rural households, questionnaire interview and participatory rural appraisal tools were used with selected farming households in three upazilas of Mymensingh district. The most prevalent diseases as reported by the farmers were red spot, followed by anal protrusion, tail and fin rot, pop eye, dropsy and gill rot. Other conditions like cotton wool type lesion, ulceration and white spot were reported but with lower incidence. Four isolates of Aeromonas hydrophila were recovered from kidney and lesion of diseased fish. Hemorrhage over the body especially near mouth and caudal region was noticed in the fishes associated with aeromonad infection. Internally, kidney, liver and spleen became swollen and enlarged. The isolates varied with their pathogenicity. All the four isolates were sensitive to Nitrofurantoin, Cotrimoxazole and Tetracycline but were resistant to Amoxycilline. An attempt was made to treat diseased fish with extracts from neem leaf, garlic and turmeric. Recovery of infection was monitored through mortality and histopathology. General histopathological changes of different organs were also studied. Extract from neem (Azadirachta indica) leaf gave better result. Telangiectasis, lamellar hypertrophy and hyperplasia hemorrhage, lamellar fusion, necrosis of lamellar epithelial cells, presence of parasites and their cysts were the major pathology of gills. Hemorrhagic lesion, pyknotic nuclei and melanomacrophage centers (MMC) were found in the liver of fish. Major pathologies in kidney of fish included presence of MMC, necrotic and ruptured kidney tubules, severe haemopoietic necrosis, and hemorrhage. The economic loss due to disease in Thai pangas farming was estimated from the difference between expected production and actual production. On an average, Thai pangas farmers of Mymensingh incur a loss of Tk. 23,104/ha/cycle due to fish disease (3.6% of expected total production). The loss, however, varied with location and size of farms, type of farmers and management practices. The study also highlighted fish health management related problems and recommended further work for the development of user-friendly farmer-oriented fish health management packages.

Deciding to evacuate: Gathering the information to make this important decision

When hazardous storms threaten coastal communities, people need information to decide how to respond to this potential emergency. NOAA and NC Sea Grant are funding a two-year project (Risk Perceptions and Emergency Communication Effectiveness in Coastal Zones) to learn how residents, government officials, businesses and other organizations are informed and use information regarding hurricane and tropical storms. (PDF contains 4 pages)