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Fish thymocyte viability, apoptosis and necrosis: In-vitro effects of organochlorine contaminants

The thymus is believed to be a central component of haematopoiesis and immune function in teleosts. Hence, chemically-elicited adverse effects to the thymus may result in immunomodulation and organ dysfunction. The objective of this research was to assess the levels of active (apoptotic) and passive (necrotic) cell death in untreated and organochlorine treated fish thymocytes. Lake trout ( Salvelinus namaycush ) thymocytes were challenged with Aroclor 1254 (concentration range 1·5–10·5 μg ml −1 ) and alpha, beta, gamma, delta isomers of hexachlorocyclohexane (concentration range 10–100 μ M ). The resulting maintenance or loss of viability was assessed by cytofluorometry (expression of phosphatidylserine and exclusion of propidium iodide) and confirmed with fluorescence microscopy. The results indicate that 20–60% of thymocytes in healthy fish undergo apoptosis, whereas thymocytes treated for 6–24 h with organochlorines exhibit increased levels of apoptotic cell death. This study demonstrates that given sufficient concentration, contact time and cellular receptors, organochlorines such as Aroclor 1254 and hexachloro-cyclohexanes may induce direct or indirect toxicity, altered functionality, or cell death to an organ important for fish immunocompetence.

Fish and Shellfish Immunology

Measurement of in vitro leucocyte mitogenesis in fish: ELISA based detection of the thymidine analogue 5'-bromo-2'-deoxyuridine

In this study we present a method for the measurement of in vitro mitogenesis in fish leucocytes that is based on the incorporation of the thymidine analogue 5′-bromo-2′-deoxyuridine (BrdU) into the DNA of replicating cells, followed by ELISA-based detection. This technique, adapted from methods developed for mammalian cells, operates on a similar biological principle to 3 H-thymidine incorporation, but circumvents the logistical and safety issues inherent with the radioactive label. Because it directly measures DNA proliferation, the assay has advantages over other colorimetric methods that may be strongly influenced by leucocyte metabolic status. Using BrdU incorporation followed by ELISA, we evaluate the responsiveness of rainbow trout ( Oncorhynchus mykiss [Walbaum]) leucocytes to the mammalian T-cell mitogen Concanavalin A (Con A) as well as the differential response of white perch ( Morone americana [Gmelin]) leucocytes to Con A and pokeweed mitogen. Specific considerations intrinsic to the assay system are discussed, including the implications of utilising enzyme-based detection.

Fish and Shellfish Immunology

Comparison of whole body and tissue blood volumes in rainbow trout (Salmo gairdneri) with 125 I bovine serum albumin and 51 Cr-erythrocyte tracers

Total, packed cell and, plasma volume estimates were made for the whole body and selected tissues of rainbow trout by the simultaneous injection of radiolabelled trout erythrocyte ( 51 Cr-RBC) and radioiodinated bovine serum albumin ( 125 I-BSA) tracers. Blood volumes were estimated with both markers separately by the tracer-hematocrit method and as the combination of the 51 Cr-RBC packed cell and 125 I-BSA plasma volumes. Mean whole body blood volume was significantly less when calculated from the 51 Cr-RBC tracer data (3.52±0.78 ml/100 g; ±SD) than when calculated with the 125 I-BSA tracer (5.06±0.86 ml/100 g) or as the sum of the two volumes combined (4.49±0.60 ml/100 g). The whole body hematocrit (28±5%), estimated as the quotient of the 51 Cr-RBC volume divided by the sum of the 125 I-BSA and the 51 Cr-RBC volumes, also was significantly less than the dorsal aortic microhematocrit (36±4%). Estimates of total blood volumes in most tissues were significantly smaller when calculated from the 51 Cr-RBC data than when calculated by the other two methods. Tissue blood volumes were greatest in highly vascularized and well perfused tissues and least in poorly vascularized tissues. The relative degree of vascularization among tissues generally remained the same regardless of whether the red cell or the plasma tracer was used to calculated blood volume. It is not clear whether the expanded plasma volume is the result of the distribution of erythrocyte-poor blood into the secondary circulation or the result of extravascular exchange of plasma proteins.

Fish Physiology and Biochemistry

An integrated data-directed numerical method for estimating the undiscovered mineral endowment in a region

An integrated data-directed numerical method has been developed to estimate the undiscovered mineral endowment within a given area. The method has been used to estimate the undiscovered uranium endowment in the San Juan Basin, New Mexico, U.S.A. The favorability of uranium concentration was evaluated in each of 2,068 cells defined within the Basin. Favorability was based on the correlated similarity of the geologic characteristics of each cell to the geologic characteristics of five area-related deposit models. Estimates of the undiscovered endowment for each cell were categorized according to deposit type, depth, and cutoff grade. The method can be applied to any mineral or energy commodity provided that the data collected reflect discovered endowment. ?? 1994 Oxford University Press.

Nonrenewable Resources

Factors affecting the toxicity of methylmercury injected into eggs

We developed a standardized protocol for comparing the sensitivities of the embryos of different bird species to methylmercury when methylmercury was injected into their eggs. During the course of developing this protocol, we investigated the effects of various factors on the toxicity of the injected methylmercury. Most of our experiments were done with chicken ( Gallus domesticus ), mallard ( Anas platyrhynchos ), and ring-necked pheasant ( Phasianus colchicus ) eggs, all of which were purchased in large numbers from game farms. A smaller amount of work was done with double-crested cormorant ( Phalacrocorax auritus ) eggs collected from the wild. Several solvents were tested, and corn oil at a rate of 1 μl/g egg contents was selected for the final standardized protocol because it had minimal toxicity to embryos and because methylmercury dissolved in corn oil yielded a dose–response curve in a range of egg concentrations that was similar to the range that causes reproductive impairment when the mother deposits methylmercury into her own eggs. The embryonic stage at which eggs were injected with corn oil altered mercury toxicity; at early stages, the corn oil itself was toxic. Therefore, in the final protocol we standardized the time of injection to occur when each species reached the morphologic equivalent of a 3-day-old chicken embryo. Although solvents can be injected directly into the albumen of an egg, high embryo mortality can occur in the solvent controls because of the formation of air bubbles in the albumen. Our final protocol used corn oil injections into the air cell, which are easier and safer than albumen injections. Most of the methylmercury, when dissolved in corn oil, injected into the air cell passes through the inner shell membrane and into the egg albumen. Most commercial incubators incubate eggs in trays with the air cell end of the egg pointing upward, but we discovered that mercury-induced mortality was too great when eggs were held in this orientation. In addition, some species of bird eggs require incubation on their sides with the eggs being rolled 180° for them to develop normally. Therefore, we adopted a procedure of incubating the eggs of all species on their sides and rolling them 180° every hour. Little has been published about the conditions of temperature, humidity, and the movements to which eggs of wild birds need to be subjected for them to hatch optimally under artificial incubation. Not unexpectedly, hatching success in an artificial incubator is generally less than what natural incubation by the parents can achieve. However, the survival of control embryos of most wild bird species was good (generally ≥ 80%) up to within 1 or 2 days of hatching when we incubated the eggs at 37.5°C (or 37.6°C for gallinaceous species) at a relative humidity that resulted in an approximate 15% to 16% loss in egg weight by the end of incubation and by incubating the eggs on their sides and rolling them 180°/h. To improve statistical comparisons, we used survival through 90% of incubation as our measurement to compare survival of controls with survival of eggs injected with graded concentrations of mercury.

Archives of Environmental Contamination and Toxico

Monitoring responses to variation in food supply for a migratory waterfowl: American Black Duck (Anas rubripes) in winter

Wintering Black Ducks ( Anas rubripes ) concentrate in wetlands along the Atlantic coast where natural and anthropogenic disturbances have increased over the last 50 years, a period in which the population of Black Ducks has declined. We studied the sensitivity of Black Ducks to perturbations in food supply that often result from disturbances by storms, predators, and people. In the paper, we characterize the responses of captive Black Ducks to shifts in food quality and availability during winter and apply those measures to a comparison of wild birds. Captive ducks that were fed intermittently (3 consecutive days/week) compensated for fasted days to achieve similar body mass and body fat to control birds that were fed every day on both animal- and plant-based diets. However, birds that were fed intermittently expended 15% more energy each day than controls when both groups were fed (536 vs. 464 kJ/kg 0.75 ), which indicates that variable food supply increases the costs of maintenance and thus reduces the number of birds that can be supported on the same resource of food without interruptions to foraging. Egg production was not affected by diet quality provided in spring or by the frequency of feeding during the preceding winter months. Black Ducks lost body fat through winter in captivity and in the wild. Fat stores of birds in New Jersey were greater than those of birds in Maine (13.3 vs. 8.3% of body mass) in January, which reflected the high energy demands of cold temperatures in Maine. Values for ∂ 15 N were greater in Maine than in New Jersey for both red blood cells and plasma, which indicated a consistent diet of marine invertebrates in Maine. Greater isotopic variation in red blood cells indicated that diets were more diverse in New Jersey than in Maine for both ∂ 15 N (9.7 ± 1.1 vs. 11.2 ± 0.4‰) and for ∂ 13 C (− 15.1 ± 2.2 vs. − 13.8 ± 1.4‰). Plasma ∂ 13 C was enriched over red blood cells in wild birds especially those with low fat stores, which suggested birds with low energy stores were shifting diets. Black Ducks can compensate for disturbances in feeding by increasing intakes if they have access to high quality wetlands where they are able to find abundant food. High energy demands at cold temperatures may constrain fat stores and thus the tolerance of feeding disturbances especially at the northern limits of the winter range. We hypothesize that decreasing variation in diet may indicate an increase in vulnerability to disturbance in winter when body fat is low. Recent efforts to assess and improve habitat quality of Black Ducks could be enhanced by monitoring the body composition and diet of birds to assess their vulnerability to disturbances in food supply and energy demands.

Journal of Comparative Physiology B: Biochemical,

Cutaneous mastocytomas in the neotenic caudate amphibians Ambystoma mexicanum (axolotl) and Ambystoma tigrinum (tiger salamander)

Spontaneous mastocytomas studied in 18 axolotls ( Ambystoma mexicanum ) and six tiger salamanders ( Ambystoma tigrinum ) were gray-white, uni- to multilobular cutaneous protrusions from 2 mm to 2 cm in diameter. Tumors were moderately cellular unencapsulated masses that usually infiltrated the dermis and hypodermis with the destruction of intervening tissues. Some tumors were invading superficial bundles of the underlying skeletal muscle. Tumors consisted of mitotically active cells derived from a single lineage but showing a range of differentiation. Immature cells had nearly smooth to lightly cleft or folded basophilic nuclei bordered by a band of cytoplasm with few cytoplasmic processes and containing a few small uniform eccentric granules. Mature cells had basophilic nuclei with deep clefts or folds and abundant eosinophilic cytoplasm with multiple long intertwining cytoplasmic extensions packed with metachromatic granules. The axolotls were old individuals from an inbred laboratory colony. The tiger salamanders were wild animals from a single polluted pond. They could have been old and inbred. Both groups were neotenic. These are the first mastocytomas discovered in cold-blooded animals.

Texas

Electricity generation by anaerobic bacteria and anoxic sediments from hypersaline soda lakes

Anaerobic bacteria and anoxic sediments from soda lakes produced electricity in microbial fuel cells (MFCs). No electricity was generated in the absence of bacterial metabolism. Arsenate respiring bacteria isolated from moderately hypersaline Mono Lake ( Bacillus selenitireducens ), and salt-saturated Searles Lake, CA (strain SLAS-1) oxidized lactate using arsenate as the electron acceptor. However, these cultures grew equally well without added arsenate using the MFC anode as their electron acceptor, and in the process oxidized lactate more efficiently. The decrease in electricity generation by consumption of added alternative electron acceptors (i.e. arsenate) which competed with the anode for available electrons proved to be a useful indicator of microbial activity and hence life in the fuel cells. Shaken sediment slurries from these two lakes also generated electricity, with or without added lactate. Hydrogen added to sediment slurries was consumed but did not stimulate electricity production. Finally, electricity was generated in statically incubated “intact” sediment cores from these lakes. More power was produced in sediment from Mono Lake than from Searles Lake, however microbial fuel cells could detect low levels of metabolism operating under moderate and extreme conditions of salt stress.

Extremophiles

Description of Tessaracoccus profundi sp.nov., a deep-subsurface actinobacterium isolated from a Chesapeake impact crater drill core (940 m depth)

A novel actinobacterium, designated CB31 T , was isolated from a 940 m depth sample of a drilling core obtained from the Chesapeake meteor impact crater. The strain was isolated aerobically on R2A medium agar plates supplemented with NaCl (20 g l -1 ) and MgCl 2 ???6H 2 O (3 g l -1 ). The colonies were circular, convex, smooth and orange. Cells were slightly curved, rod-shaped in young cultures and often appeared in pairs. In older cultures cells were coccoid. Cells stained Gram-positive, were non-motile and did not form endospores. The diagnostic diamino acid of the peptidoglycan was ll-diaminopimelic acid. The polar lipids included phosphatidylglycerol, diphosphatidglycerol, four different glycolipids, two further phospholipids and one unidentified lipid. The dominant menaquinone was MK-9(H 4 ) (70%). The major cellular fatty acid was anteiso C15:0 (83%). The DNA G + C content was 68 mol%. The strain grew anaerobically by reducing nitrate to nitrite or by fermenting glucose. It was catalase positive and oxidase negative. It grew between 10 and 45??C, with an optimum between 35 and 40??C. The pH range for growth was 5.7-9.3, with an optimum at pH 7.5. The closest phylogenetic neighbors based on 16S rRNA gene sequence identity were members of the genus Tessaracoccus (95-96% identity). On the basis of phenotypic and phylogenetic distinctiveness, strain CB31 T is considered to represent a novel species of the genus Tessaracoccus, for which we propose the name Tessaracoccus profundi sp. nov.. It is the first member of this genus that has been isolated from a deep subsurface environment. The type strain is CB31 T (=NCIMB 14440 T = DSM 21240 T ). ?? 2009 Springer Science+Business Media B.V.

Antonie van Leeuwenhoek

Effects of dietary exposure to brominated flame retardant BDE-47 on thyroid condition, gonadal development and growth of zebrafish

Little is known about the effects of brominated flame retardants in teleosts and some of the information currently available is inconsistent. This study examined effects of dietary exposure to 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) on thyroid condition, body mass and size, and gonadal development of zebrafish. Pubertal, 49-day-old (posthatch) fish were fed diets without BDE-47 (control) or with 1, 5 or 25 μg/g BDE-47/diet. Treatments were conducted in triplicate 30-L tanks each containing 50 zebrafish, and 15 fish per treatment (5 per tank) were sampled at days 40, 80 and 120 of exposure. Measurements were taken of body mass, standard length, head depth and head length. Sex (at 40–120 days of exposure), germ cell stage (at 40 days) and thyroid condition (at 120 days; follicular cell height, colloid depletion, angiogenesis) were histologically determined. Whole-body BDE-47 levels at study completion were within the high end of levels reported in environmentally exposed (wild) fishes. Analysis of variance was used to determine differences among treatments at each sampling time. No effects were observed on thyroid condition or germ cell stage in either sex. Reduced head length was observed in females exposed to BDE-47 at 80 days but not at 40 or 120 days. In males, no apparent effects of BDE-47 were observed at 40 and 80 days, but fish exposed to 25 μg/g had lower body mass at 120 days compared to control fish. These observations suggest that BDE-47 at environmentally relevant whole-body concentrations does not affect thyroid condition or pubertal development of zebrafish but does affect growth during the juvenile-to-adult transition, especially in males.

Fish Physiology and Biochemistry

A direct immunoassay for detecting diatoms in groundwater as an indicator of the direct influence of surface water

Groundwater under the direct influence of surface water (GWUDISW) is of concern in communities where growing public demand on groundwater resources has resulted in increased withdrawals and hydraulic stress near surface water bodies. Under these conditions, contaminants such as methyl-tert butyl ether (MTBE) and biological materials have been detected in domestic wells. Other contaminants and pathogens associated with surface water are not routinely tested for in groundwater-supplied systems. To address the need for methods to easily identify potentially vulnerable supplies, a direct immunoassay for the quantitative detection of diatoms in raw water samples was developed as a measure of surface water influence on groundwater. Cell wall preparations from Nitzschia palea Kützing, a freshwater diatom found throughout North America, were used to produce a polyclonal antibody that was applied in a direct enzyme-linked immunosorbent assay (ELISA) developed to detect the presence of N. palea cell wall components. The direct immunoassay allows detection at 500 cells L −1 , a level similar to diatom concentrations observed in samples of groundwater collected near the test site. This investigation was the first attempt to utilize an ELISA as an indicator of surface water influence on groundwater. Further research is needed to develop more specific diatom-based monoclonal antibodies, determine cross-reactivity, and optimize sample processing and ELISA procedures for development of a standardized method.

Journal of Applied Phycology

Empirical methods for detecting regional trends and other spatial expressions in antrim shale gas productivity, with implications for improving resource projections using local nonparametric estimation techniques

The primary objectives of this research were to (1) investigate empirical methods for establishing regional trends in unconventional gas resources as exhibited by historical production data and (2) determine whether or not incorporating additional knowledge of a regional trend in a suite of previously established local nonparametric resource prediction algorithms influences assessment results. Three different trend detection methods were applied to publicly available production data (well EUR aggregated to 80-acre cells) from the Devonian Antrim Shale gas play in the Michigan Basin. This effort led to the identification of a southeast–northwest trend in cell EUR values across the play that, in a very general sense, conforms to the primary fracture and structural orientations of the province. However, including this trend in the resource prediction algorithms did not lead to improved results. Further analysis indicated the existence of clustering among cell EUR values that likely dampens the contribution of the regional trend. The reason for the clustering, a somewhat unexpected result, is not completely understood, although the geological literature provides some possible explanations. With appropriate data, a better understanding of this clustering phenomenon may lead to important information about the factors and their interactions that control Antrim Shale gas production, which may, in turn, help establish a more general protocol for better estimating resources in this and other shale gas plays.

Michigan

Beat-the-wave evacuation mapping for tsunami hazards in Seaside, Oregon, USA

Previous pedestrian evacuation modeling for tsunamis has not considered variable wave arrival times or critical junctures (e.g., bridges), nor does it effectively communicate multiple evacuee travel speeds. We summarize an approach that identifies evacuation corridors, recognizes variable wave arrival times, and produces a map of minimum pedestrian travel speeds to reach safety, termed a “beat-the-wave” (BTW) evacuation analysis. We demonstrate the improved approach by evaluating difficulty of pedestrian evacuation of Seaside, Oregon, for a local tsunami generated by a Cascadia subduction zone earthquake. We establish evacuation paths by calculating the least cost distance (LCD) to safety for every grid cell in a tsunami-hazard zone using geospatial, anisotropic path distance algorithms. Minimum BTW speed to safety on LCD paths is calculated for every grid cell by dividing surface distance from that cell to safety by the tsunami arrival time at safety. We evaluated three scenarios of evacuation difficulty: (1) all bridges are intact with a 5-minute evacuation delay from the start of earthquake, (2) only retrofitted bridges are considered intact with a 5-minute delay, and (3) only retrofitted bridges are considered intact with a 10-minute delay. BTW maps also take into account critical evacuation points along complex shorelines (e.g., peninsulas, bridges over shore-parallel estuaries) where evacuees could be caught by tsunami waves. The BTW map is able to communicate multiple pedestrian travel speeds, which are typically visualized by multiple maps with current LCD-based mapping practices. Results demonstrate that evacuation of Seaside is problematic seaward of the shore-parallel waterways for those with any limitations on mobility. Tsunami vertical-evacuation refuges or additional pedestrian bridges may be effective ways of reducing loss of life seaward of these waterways.

Oregon

Delta-aminolevulinic acid dehydratase enzyme activity in blood, brain, and liver of lead-dosed ducks

Mallard ducks were dosed with a single shotgun pellet (ca. 200 mg lead). After 1 month there was about 1 ppm lead in blood, 2.5 in liver, and 0.5 in brain. Lead-induced inhibition of delta-aminolevulinic acid dehydratase enzyme in blood and cerebellum was much greater than in cerebral hemisphere or liver and was strongly correlated with the lead concentration in these tissues. The cerebellar portion of the brain was more sensitive to delta-aminolevulinic acid dehydratase enzyme inhibition by lead than were the other tissues examined. There was also a greater increase in the glial cell marker enzyme, butyrylcholinesterase, in cerebellum than in cerebral hemisphere, suggesting that nonregenerating neuronal cells were destroyed by lead and replaced by glial cells in that portion of the brain. Even partial loss of cerebellar tissue is severely debilitating in waterfowl, because functions critical to survival such as visual, auditory, motor, and reflex responses are integrated at this brain center.

Environmental Research

Potencies of polychlorinated dibenzo-p-dioxin, dibenzofuran, and biphenyl congeners, relative to 2,3,7,8-tetrachlorodibenzo-p-dioxin, for producing early life stage mortality in rainbow trout (Oncorhynchus mykiss)

The presence of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs) in feral lake trout eggs ( Salvelinus namaycush ) may increase the risk of lake trout early life stage mortality in the Great Lakes. To assess the combined toxicity of PCDDs, PCDFs, and PCBs to lake trout early development, toxic potencies, relative to 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), were determined for individual PCDD, PCDF, and PCB congeners, based on rainbow trout ( Oncorhynchus mykiss ) early life stage mortality. Newly fertilized rainbow trout eggs were injected with graded doses of 2,3,7,8-TCDD, or a PCDD, PCDF, or PCB congener. LD 50 values were determined based on the egg dose that caused mortality from hatching onset to swim-up. Toxic equivalency factors (TEFs) were calculated as 2,3,7,8-TCDD LD 50 /congener LD 50 . TEFs were for PCDDs: 2,3,7,8-TCDD = 1.0; 1,2,3,7,8-PeCDD = 0.730;and 1,2,3,4,7,8-HxCDD = 0.319; for PCDFs: 2,3,4,7,8-PeCDF = 0.359; 1,2,3,4,7,8-HxCDF = 0.280; 1,2,3,7,8-PeCDF = 0.034; and 2,3,7,8-TCDF = 0.028; and for PCBs: 3,3',4,4',5-PeCB = 0.005; 3,3',4,4'-TCB = 0.00016; 2,3,3',4,4'-PeCB and 2,3',4,4',5-PeCB < 0.00007. For PCDDs, fish-specific TEFs were 10- to 100-fold higher than TEFs determined in H4IIE rat hepatoma cells, but were similar to TEFs proposed for risk assessment. For PCDFs, fish specific TEFs were similar to TEFs determined in H4IIE cells and TEFs proposed for risk assessment. However, the most significant finding was that for the coplanar PCBs and mono- ortho -chlorinated analogues of the coplanar PCBs fish-specific TEFs were 1/14 to 1/80 less than both those determined in H4IIE cells and proposed for risk assessment. Using these fish-specific TEFs, the risk associated with exposure of early life stages of Sake trout to complex mixtures of PCDDs, PCDFs, and PCBs in the Great Lakes can be estimated.

Aquatic Toxicology

Demethylation and cleavage of dimethylsulfoniopropionate in marine intertidal sediments

Demethylation and cleavage of dimethylsulfoniopropionate (DMSP) was measured in three different types of,intertidal marine sediments: a cyanobacterial mat, a diatom-covered tidal flat and a carbonate sediment. Consumption rates of added DMSP were highest in cyanobacterial mat slurries (59 ?? mol DMSP l-1 slurry h-1) and lower in slurries from a diatom mat and a carbonate tidal sediment (24 and 9 ??mol DMSP l-1 h-1, respectively). Dimethyl sulfide (DMS) and 3-mercaptopropionate (MPA) were produced simultaneously during DMSP consumption, indicating that cleavage and demethylation occurred at the same time. Viable counts of DMSP-utilizing bacteria revealed a population of 2 x 107 cells cm-3 sediment (90% of these cleaved DMSP to DMS, 10% demethylated DMSP to MPA) in the cyanobacterial mat, 7 x 105 cells cm-3 in the diatom mat (23% cleavers, 77% demethylators), and 9 x 104 cells cm-3 (20% cleavers and 80% demethylators) in the carbonate sediment. In slurries of the diatom mat, the rate of MPA production from added 3-methiolpropionate (MMPA) was 50% of the rate of MPA formation from DMSP. The presence of a large population of demethylating bacteria and the production of MPA from DMSP suggest that the demethylation pathway, in addition to cleavage, contributes significantly to DMSP consumption in coastal sediments.

FEMS Microbiology Ecology

Southeast Atlantic marine and terrestrial response to middle Pliocene climate change

Analysis of planktic foraminifers and pollen from Deep Sea Drilling Project Site 532 located on the continental margin of southwest Africa provides information on the link between Pliocene paleoceanographic conditions and paleoenvironments of southwest Africa. Increased upwelling at Site 532 correlates with southward migration of onshore vegetation regions. Both terrestrial and marine changes can be explained by changes in the southern hemisphere surface temperature gradient which affected the paleo-position of subtropical high pressure cells and productivity of water masses during the Pliocene. When the subtropical high pressure cells were further south, Southern Ocean waters were warmer, contained less sea ice, Southern Ocean diatom productivity was high, and nutrient depleted water upwelled off southwest Africa. When the subtropical high pressure cells were in a configuration similar to the present, Southern Ocean waters were cooler and contained more sea ice, the aerial extent of Southern Ocean diatom productivity was limited, and nutrient rich waters upwelled off southwest Africa as the Benguela upwelling system migrated closer to the position of Site 532.

Marine Micropaleontology

Weed-suppressive bacteria effects differ in culture compared to in soils and with or without microbial competition and separation of active ingredient

Weed-suppressive bacteria (WSB), specifically the D7 and ACK55 strains of Pseudomonas fluorescens , were previously reported to selectively inhibit growth of invasive annual grasses (IAGs) that have caused severe ecosystem degradation across much of the western US. However, recent studies show highly mixed evidence for WSB effectiveness in field settings. We evaluated how the effectiveness of WSB, specifically selective inhibition of exotic annual but not native perennial grasses, was altered by (1) growth media (soil vs. agar culture), (2) pre-sterilization to remove competing microbes, and (3) WSB concentration, reduced by centrifuging with and without filtration. WSB had no effects in soil microcosms with or without pre-sterilization, but WSB did have effects in agar cultures. On agar, WSB effects were amplified by pre-sterilizing seeds prior to treatment with WSB inoculum – but effects were non-selective (i.e., native and exotic plants were equally inhibited) with or without pre-sterilization at high WSB concentration. The only selective effects of WSB were observed on agar at low WSB concentrations, made possible by centrifuging WSB cells out of the inoculum prior to treatment application (from ~10 7 to ~10 5 live cells/ml). Completely removing WSB cells from inoculum by filtration led to no WSB effects. While these results suggest the desired effects of WSB can be reproduced on agar with a supernatant containing the putative active agent and a low density of WSB cells, the effect is still not reproducible for plants in soil, even when competing microbes are removed. Thus, additional research is needed to determine if, when, where, and how WSB could be effective in field settings.

Biological Control