Oil spill remediation by using the remediation agent JE1058BS that contains a biosurfactant produced by Gordonia sp. strain JE-1058

A remediation agent containing a biosurfactant was prepared by spray drying the sterilized culture broth of Gordonia sp. strain JE-1058, and the agent was designated as JE1058BS. On subjection to the baffled flask test developed by the United States Environmental Protection Agency, JE1058BS showed a strong potential to be applied as an oil spill dispersant even in the absence of a solvent. It also proved to be an effective bioremediation agent for the remediation of oil spills at sea. The addition of JE1058BS to seawater stimulated the degradation of weathered crude oil (ANS 521) via the activity of the indigenous marine bacteria. Its addition also stimulated the removal of crude oil from the surface of contaminated sea sand. These results indicate that biosurfactant-containing JE1058BS has a strong potential to be applied as a remediation agent for the clean-up of oil spills at sea and on shorelines.     

Reconciling Biodiversity with Fishing: A Holistic Strategy for Pacific Sea Turtle Recovery

Recovery of sea turtle populations requires addressing: multiple sources of mortality; nonmarket, diffuse benefits with costs localized on the poor; and a transboundary resource with incomplete jurisprudence, markets, and institutions. Holistic recovery strategies include: beach conservation protecting nesting females, their eggs, and critical breeding habitat to maximize hatchling production; enhanced at-sea survival of turtles on the high seas and in commercial coastal fisheries; and reduced artisanal coastal fisheries mortality of turtles. The traditional approach of focusing long-term sustained conservation efforts on the nesting beaches has by itself led to increases in several sea turtle populations. However, current conservation is inadequate to reverse declines in other cases such as the critically endangered leatherback populations in the Pacific. This article discusses policy instruments comprising a holistic recovery strategy that reconciles fishing with biodiversity conservation.     

Morphopathological findings in dog's acute toxoplasmosis

Toxoplasmosis was clinically diagnosed in two dogs of 4 and respectively 18 months. A seven-day treatment remained inefficient and the animals died. Autopsy revealed a global inflammation of the lung, with necrotic lesions of bronchial lymph nodes and acute hyperplastic reaction of the spleen. Histologically, there were identified a diffuse, serofibrinous inflammation of the lung with necrotic foci, fibrinoleucocytic and necrotic exudate of bronchial and portal lymph nodes, small necrotic or fibrinonecrotic foci of the liver and spleen, lymphoglial foci and edema at the base of the cerebellum. In myocardium, as well as in the above mentioned organs, toxoplamas are free or accumulated in cysts. Inoculation of infected lung material into mice and newborn dogs gave positive results.     

Some analyses of artificially incubated eggs and hatchlings of Green and Loggerhead sea turtles

Eggs of the Green sea turtle ( Chelonia mydas ) and the Loggerhead sea turtle ( Caretta caretta ) were incubated in coral sand or silica sand moistened with distilled water. The eggs and hatchlings were analysed for calcium, magnesium and phosphorus. There was less magnesium but more calcium in the hatchlings than in the original egg contents but this was not influenced by the type of sand used during incubation. The results are interpreted as indicating that the eggshell is the major source of calcium for the developing embryos. The Logger-head sea turtle shows a greater efficiency in the incorporation of yolk magnesium and phosphorus into the embryo than does the Green sea turtle.     

Hazards Associated with the Consumption of Sea Turtle Meat and Eggs: A Review for Health Care Workers and the General Public

Sea turtle products (e.g., meat, adipose tissue, organs, blood, eggs) are common food items for many communities worldwide, despite national regulations in some countries prohibiting such consumption. However, there may be hazards associated with this consumption due to the presence of bacteria, parasites, biotoxins, and environmental contaminants. Reported health effects of consuming sea turtles infected with zoonotic pathogens include diarrhea, vomiting, and extreme dehydration, which occasionally have resulted in hospitalization and death. Levels of heavy metals and organochlorine compounds measured in sea turtle edible tissues exceed international food safety standards and could result in toxic effects including neurotoxicity, kidney disease, liver cancer, and developmental effects in fetuses and children. The health data presented in this review provide information to health care providers and the public concerning the potential hazards associated with sea turtle consumption. Based on past mortality statistics from turtle poisonings, nursing mothers and children should be particularly discouraged from consuming all sea turtle products. We recommend that individuals choose seafood items lower in the food chain that may have a lower contaminant load. Dissemination of this information via a public health campaign may simultaneously improve public health and enhance sea turtle conservation by reducing human consumption of these threatened and endangered species.     

Relationship between tropical cyclones and the distribution of sea turtle nesting grounds

Aim This study examines the relationship between the distribution of existing sea turtle nesting sites and historical patterns of tropical cyclone events to investigate whether cyclones influence the current distribution of sea turtle nesting sites. The results, together with information on predicted cyclone activity and other key environmental variables, will help in the identification and prediction of future nesting sites for sea turtles as changes to the coastal environment continue. Location Queensland, Australia. Methods We used data on the nesting distribution of seven populations of four species of sea turtles [green (Chelonia mydas), flatback (Natator depressus), hawksbill (Eretmochelys imbricata) and loggerhead (Caretta caretta)] from the eastern Queensland coast, and tropical cyclone track data from 1969 to 2007 to explore the relationship between (1) sea turtle nesting phenology and cyclone season, and (2) sea turtle nesting sites and cyclone distribution. Furthermore, using two green turtle populations as a case study, we investigated the relationship between cyclone disturbance and sea turtle reproductive output, nesting site and season. Bootstrapping was used to explore if current sea turtle nesting sites are located in areas with lower or higher cyclone frequency than areas where turtles are currently not nesting. Results All populations of sea turtles studied here were disturbed by cyclone activity during the study period. The exposure (frequency) of tropical cyclones that crossed each nesting site varied greatly among and within the various sea turtle populations. This was mainly a result of the spatial distribution of each population’s nesting sites. Bootstrapping indicated that nesting sites generally have experienced lower cyclone activity than other areas that are available for nesting. Main conclusions Tropical cyclones might have been sufficiently detrimental to sea turtle hatching success on the eastern Queensland coast that through a natural selection process turtles in this region are now nesting in areas with lower cyclone activity. Therefore, it is important that future studies that predict climate or range shifts for sea turtle nesting distributions consider future cyclone activity as one of the variables in their model.     

Highly efficient Aerococcus viridans L-α-glycerophosphate oxidase production in the presence of H2O2-decomposing agent: purification and kinetic characterization

Glycerophosphate oxidase was purified from Aerococcus viridans cells by phase partitioning in Triton X-114, ammonium sulfate fractionation, FPLC ion-exchange chromatography and FPLC hydrophobic-interaction chromatography. The purification achieved from a crude extract of A. viridans was 38-fold with a 32% recovery of activity. Under the growth conditions used, A. viridans strain CECT 978 proved to be an excellent glycerophosphate-oxidase producer, with enzyme production 2,800-fold greater than that described in the literature for the same microorganism. The culture medium used in the present work is that commonly used for cultivation of this microorganism, except that an H2O2-decomposing enzyme was added. The addition of catalase to the growth medium had a clear effect on the growth rate. Furthermore, methylglyoxal, a metabolite that is formed enzymatically from triose phosphates, was found to be an inactivator of glycerophosphate oxidase activity.     

Differential Gene Expression Induced by Exposure of Captive Mink to Fuel Oil: A Model for the Sea Otter

Free-ranging sea otters are subject to hydrocarbon exposure from a variety of sources, both natural and anthropogenic. Effects of direct exposure to unrefined crude oil, such as that associated with the Exxon Valdez oil spill, are readily apparent. However, the impact of subtle but pathophysiologically relevant concentrations of crude oil on sea otters is difficult to assess. The present study was directed at developing a model for assessing the impact of low concentrations of fuel oil on sea otters. Quantitative PCR was used to identify differential gene expression in American mink that were exposed to low concentrations of bunker C fuel oil. A total of 23 genes, representing 10 different physiological systems, were analyzed for perturbation. Six genes with immunological relevance were differentially expressed in oil-fed mink. Interleukin-18 (IL-18), IL-10, inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), and complement cytolysis inhibitor (CLI) were down-regulated while IL-2 was up-regulated. Expression of two additional genes was affected; heat shock protein 70 (HSP70) was up-regulated and thyroid hormone receptor (THR) was down-regulated. While the significance of each perturbation is not immediately evident, we identified differential expression of genes that would be consistent with the presence of immune system-modifying and endocrine-disrupting compounds in fuel oil. Application of this approach to identify effects of petroleum contamination on sea otters should be possible following expansion of this mink model to identify a greater number of affected genes in peripheral blood leukocytes.     

Sequential Growth of Bacteria on Crude Oil

By modification of the enrichment culture procedure three bacterial strains capable of degrading crude oil in sea water were isolated in pure culture, UP-2, UP-3, and UP-4. Strain UP-2 appears to be highly specialized for growth on crude oil in sea water since it showed strong preference for oil or oil degradation products as substrates for growth, converted 66% of the oil into a form no longer extractable by organic solvents, quantitatively degraded the paraffinic fraction (gas chromatographic analysis), emulsified the oil during exponential growth, and produced 1.6 × 10⁸ cells per mg of oil. After exhaustive growth of UP-2 on crude oil the residual oil supported the growth of UP-3 and UP-4, but not a previously isolated oil-degrading bacterium, RAG-1. Strains UP-2, UP-3, and UP-4 grew on RAG-1-degraded oil (specifically depleted of n-alkanes). The growth of UP-3 and UP-4 on UP-2 and RAG-1-degraded oil resulted in the production of new paraffinic compounds as revealed by gas chromatography. When the four strains were grown either together in a mixed culture or sequentially, there was over 75% oil conversion. By plating on selective media, growth of the individual strains was measured kinetically in the reconstituted mixed culture, revealing competition for common growth substances (UP-2 and RAG-1), enhanced die-off (UP-2), and stabilization (UP-4) during the stationary phase.     

仿野生鳖卵的磷脂及粗蛋白含量测定

仿野生鳖卵内容物冻干粉收率为 30 .6 1% ,它的粗蛋白质及磷脂含量分别为 5 0 .0 1%和 12 .74 %。TLC分析结果表明仿野生鳖卵含有 5种磷脂成份 ,而鹌鹑卵及猪脑仅含有 3种磷脂     

Use of dye affinity chromatography for the purification of Aerococcus viridans lactate oxidase

Lactate oxidase was purified from Aerococcus viridans (A. viridans) by dye affinity chromatography and FPLC ion exchange chromatography. The lactate oxidase could be purified by comparatively simple procedures, the purification achieved from a crude extract of A. viridans was 41-fold with a specific activity of 143 units/(mg of protein). The purified enzyme was a L-lactate oxidase, which catalyses the conversion of L-lactate in the presence of molecular oxygen to pyruvate and H(2)O(2). This purified lactate oxidase showed an apparent molecular mass of 48,200 in SDS-PAGE and the native molecular weight, as estimated by FPLC gel filtration, was 187,300. This molecular weight indicates that lactate oxidase exists in tetrameric form after gel filtration. To differing degrees, all the triazine dyes tested were inhibitors of lactate oxidase, solutions of free triazine dyes showing an inhibition mechanism which was both time- and pH-dependent.     

太平洋丽龟的生理结构研究

目的研究太平洋丽龟的生理结构及特点,为我国开展海龟保育和科学研究提供参考。方法对1只误捕致死的太平洋丽龟进行解剖,观察其循环系统、消化系统、呼吸系统和泄殖系统并测量记录。结果太平洋丽龟心脏位于胸部正中、稍偏右;食管内壁分布许多锥状突起;小肠长240cm,占整个消化系统管道总长的53.1%,消化道总长与体长的比值（简称肠长比）为5.35;精巢1对,阴茎长30cm,可推断为成年的雄龟。结论太平洋丽龟的生理结构估计与其肉食性有关,这对开展人工驯养繁育和物种保护具有重要意义。     

Sea Turtle Nesting as a Process Influencing a Sandy Beach Ecosystem

Sea turtle egg mortality, egg predation, and small organisms associated with turtle nests were studied at Playa Ostional, Costa Rica. Sites with concentrated sea turtle nesting were compared with solitary nesting sites as a function of place and time based on ANOVA, Akaike's Information Criterion, and Bayesian analyses. Results indicate that sea turtle egg mortality was significantly associated (P < 0.005) with flowing water that erodes or saturates nesting sites, and with overlapped nesting in which sea turtles disturb each other's nests. Sarcophagid and calliphorid fly larvae (Bayesian prior = 1.19; posterior = 2.27), fungi (prior = 1.14; posterior = 1.92), mites (prior = 0.51; posterior = 1.15), and several other types of small organisms increased in number after turtle egg laying (N= 303 nests; 34,451 turtle eggs). During peak sea turtle nesting periods, visitation to nesting sites by poachers and vertebrate predators was high, and relative number of nests disturbed by these predators was low (P < 0.02). In multimodel analysis, the three most parsimonious models were: (1) turtle egg mortality and distance from mean high tide; (2) turtle egg predation and distance from mean high tide; and (3) turtle egg mortality and nesting density, with Akaike weights of 0.224, 0.203, and 0.153 respectively. Intensive sea turtle nesting might result in upwelling and turnover of nesting debris and nest organisms, and may influence biotic community structure of sandy beach ecosystems.     

Microbial degradation of oil spills enhanced by a slow-release fertilizer.

The improved cleanup of marine oil spills by stimulating biodegradation through the use of a slow-release fertilizer is reported. A paraffin-supported fertilizer containing MgNH4PO4 as active ingredient was developed and evaluated in laboratory and field experiments using quantitative infrared spectrometry and chromatographic techniques. The biodegradation of Sarir crude oil in the sea was considerably enhanced by paraffin-supported fertilizer. After 21 days 63% had disappeared as compared to 40% in the control area.     

Microbial degradation of oil spills enhanced by a slow-release fertilizer.

The improved cleanup of marine oil spills by stimulating biodegradation through the use of a slow-release fertilizer is reported. A paraffin-supported fertilizer containing MgNH4PO4 as active ingredient was developed and evaluated in laboratory and field experiments using quantitative infrared spectrometry and chromatographic techniques. The biodegradation of Sarir crude oil in the sea was considerably enhanced by paraffin-supported fertilizer. After 21 days 63% had disappeared as compared to 40% in the control area.     

中华鳖4个品系营养成分分析与比较

为了解墨鳖的营养价值, 对墨鳖、野生中华鳖、淮河鳖和日本鳖肌肉的一般营养成分、氨基酸含量、脂肪酸含量及裙边胶原蛋白进行测定和比较。结果显示: (1) 4个品系鳖肌肉水分含量分别为80.84%、79.48%、81.57%和79.25%, 墨鳖显著高于野生鳖和日本鳖(P<0.05); 粗蛋白含量分别是17.82%、17.60%、15.93%和16.40%, 墨鳖含量最高, 显著高于淮河鳖和日本鳖(P<0.05); 粗脂肪含量分别是0.42%、1.43%、0.65%和1.06%, 墨鳖显著低于其他鳖(P<0.05); 4个品系鳖灰分含量差异不显著(P>0.05)。(2) 4个品系鳖肌肉中总氨基酸和呈味氨基酸含量均以墨鳖最高。(3)4个品系鳖不饱和脂肪酸含量分别为65.19%、56.44%、59.32%和54.73%, 墨鳖显著高于其他鳖(P<0.05), 其中墨鳖单不饱和脂肪酸含量显著低于野生鳖(P<0.05), 多不饱和脂肪酸含量显著高于其他3个品系(P<0.05); 饱和脂肪酸含量分别为32.50%、41.85%、39.41%和39.98%, 墨鳖显著低于其他鳖(P<0.05)。(4)墨鳖肌肉中DHA和EPA含量与野生鳖差异不显著(P>0.05), 显著高于淮河鳖和日本鳖(P<0.05), 花生四烯酸(AA)含量显著高于其他3个品系鳖(P<0.05)。(5) 4个品系鳖裙边胶原蛋白含量均较高, 为160.6—170.4 mg/g。结果表明墨鳖是一种营养价值高、肉味鲜美的地方品系。     

Effects of petroleum on mink applied as a model for reproductive success in sea otters.

Ranch-reared mink (Mustela vison) were used as a model in an experimental trial to investigate the potential effects of exposure to two petroleum products on sea otters (Enhydra lutris). Mink were exposed either dermally on one occasion 60 days prior to breeding or via low level contamination of their diets daily from 60 days prior to breeding (January 1994) until weaning of kits (June 1994). For dermal exposure, we placed mink in either a slick of Alaskan North Slope crude oil (n = 24) or bunker C fuel oil (n = 24) on sea water or sea water alone (n = 10) for 1 min. For dietary exposure, we fed mink rations containing 500 ppm of either Alaskan North Slope crude oil (n = 24) or bunker C fuel oil (n = 24; control, n = 15). The number of liveborn kits did not differ significantly among mink exposed dermally (5.0 kits/female for crude oil and 6.5 kits/female for bunker C fuel oil) and unexposed controls (5.3 kits/female). However, only 2.3 and 0.7 kits were produced per female for those exposed through the diet to crude oil and bunker C fuel oil, respectively. Females with reduced reproductive success had no clinical signs of toxicosis or behavioral abnormalities. In addition, kits of females exposed through the diet had poor survival to weaning. Once mature, kits born to females exposed to bunker C fuel oil in the diet had significantly reduced reproductive success (3.4 kits/female) although their only exposure to the petroleum products was in utero or during nursing. Therefore, it is possible that sea otter populations consuming contaminated food sources or colonizing previously oiled habitats will have reduced reproductive success.     

Influence of sea turtle nesting on hunting behavior and movements of jaguars in the dry forest of northwest Costa Rica

Jaguars (Panthera onca) are opportunistic predators that prey on large profitable prey items, such as sea turtles at nesting beaches. Here, we use jaguar and sea turtle track-count surveys, combined with satellite telemetry of one jaguar, to evaluate whether jaguar hunting behavior and movements are influenced by seasonal sea turtle nesting in the Sector Santa Rosa of Área de Conservación Guanacaste in northwest Costa Rica. We used generalized linear models to evaluate the effect of moon phase and sea surface temperature on olive ridley (Lepidochelis olivacea) and green turtle (Chelonia mydas) nesting abundance, as well as the combination of these predictors on the frequency of jaguar predation activity (proximity to nesting beaches) and movements. For home-range size and location analyses, we calculated kernel density estimates for each season at three different temporal scales. Sea turtle nesting season influenced jaguar activity patterns, as well as sea turtle abundance was related to jaguar locations and predation events, but jaguar home-range size (88.8 km² overall) did not differ between nesting seasons or among temporal scales. Environmental conditions influenced sea turtle nesting and, as a consequence, also influenced jaguar movements and foraging activity. Our study defined the home range of a female jaguar in the tropical dry forest and its relationship to seasonally abundant turtles. Additional information related to the effect of tourism on jaguar–sea turtle interactions would improve conservation of these species at unique nesting beaches in the area.     

Efficient establishment of primary fibroblast cultures from the hawksbill sea turtle (Eretmochelys imbricata)

The hawksbill sea turtle (Eretmochelys imbricata) is a critically endangered species at a risk of extinction. Preservation of the genomic and cellular information of endangered animals is important for future genetic and biological studies. Here, we report the efficient establishment of primary fibroblast cultures from skin tissue of the hawksbill sea turtle. We succeeded in establishing 19 primary cultures from 20 hawksbill sea turtle individuals (a success rate of 95%). These cells exhibited a fibroblast-like morphology and grew optimally at a temperature of 26°C, but experienced a loss of viability when cultured at 37°C. Chromosomal analysis using the primary cells derived here revealed that hawksbill sea turtles have a 2n?=?56 karyotype. Furthermore, we showed that our primary cell cultures are free of several fish-related viruses, and this finding is important for preservation purposes. To our knowledge, this report is the first to describe primary cell cultures established from normal tissues of the hawksbill sea turtle. The results will contribute to the preservation of biodiversity, especially for the sea turtles that are critically endangered owing to human activities.     

Degradation and mineralization of petroleum by two bacteria isolated from coastal waters

Within the framework of a study on the oil biodegradation potential of the sea the ability of a Flavobacterium sp. and Brevibacterium sp. to metabolize a paraffinic crude oil and a chemically defined hydrocarbon mixture was investigated. Major components of the crude oil were identified by combination gas chromatography and mass spectrometry. The rate and extent of total hydrocarbon biodegradation was measured. In addition, CO₂ evolution from the crude oil was continuously monitored in a shaker-mounted gas train arrangement. Degradation started after a 2 to 4 day lag period, and reached its maximum within two weeks. At this time up to 60% of the crude oil and 75% of the model hydrocarbon mixture, each added at the level of 1 ml per 100 ml artificial sea water, were degraded. Mineralization(conversion to CO₂) was slightly lower due to formation of products and bacterial cell material. n-Paraffins were preferentially degraded as compared to branched chain hydrocarbons. Biodegradation of n-paraffins in the range of C₁₂ to C₂₀ was simultaneous; no diauxie effects were observed.