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Accounting for intracell flow in models with emphasis on water table recharge and stream-aquifer interaction: 2. A procedure

Intercepted intracell flow, especially if cell includes water table recharge and a stream ((sink), can result in significant model error if not accounted for. A procedure utilizing net flow per cell ( F n ) that accounts for intercepted intracell flow can be used for both steady state and transient simulations. Germane to the procedure is the determination of the ratio of area of influence of the interior sink to the area of the cell ( A i / A c ). A i is the area in which water table recharge has the potential to be intercepted by the sink. Determining A i / A c requires either a detailed water table map or observation of stream conditions within the cell. A proportioning parameter M , which is equal to 1 or slightly less and is a function of cell geometry, is used to determine how much of the water that has potential for interception is intercepted by the sink within the cell. Also germane to the procedure is the determination of the flow across the streambed ( F s ), which is not directly a function of cell size, due to difference in head between the water level in the stream and the potentiometric surface of the aquifer underlying the streambed. The use of F n for steady state simulations allows simulation of water levels without utilizing head-dependent or constant head boundary conditions which tend to constrain the model-calculated water levels, an undesirable result if a comparison of measured and calculated water levels is being made. Transient simulations of streams usually utilize a head-dependent boundary condition and a leakance value to model a stream. Leakance values for each model cell can be determined from a steady state simulation, which used the net flow per cell procedure. For transient simulation, F n would not include F s . Also, for transient simulation it is necessary to check F n at different time intervals because M and A i / A c are not constant and change with time. The procedure was used successfully in two different models of the aquifer system in the Ozarks. The use of F n was essential to the two model studies because most model cells in both models contained water table recharge and multiple sinks.

Water Resources Research↗

How processing digital elevation models can affect simulated water budgets

For regional models, the shallow water table surface is often used as a source/sink boundary condition, as model grid scale precludes simulation of the water table aquifer. This approach is appropriate when the water table surface is relatively stationary. Since water table surface maps are not readily available, the elevation of the water table used in model cells is estimated via a two-step process. First, a regression equation is developed using existing land and water table elevations from wells in the area. This equation is then used to predict the water table surface for each model cell using land surface elevation available from digital elevation models (DEM). Two methods of processing DEM for estimating the land surface for each cell are commonly used (value nearest the cell centroid or mean value in the cell). This article demonstrates how these two methods of DEM processing can affect the simulated water budget. For the example presented, approximately 20% more total flow through the aquifer system is simulated if the centroid value rather than the mean value is used. This is due to the one-third greater average ground water gradients associated with the centroid value than the mean value. The results will vary depending on the particular model area topography and cell size. The use of the mean DEM value in each model cell will result in a more conservative water budget and is more appropriate because the model cell water table value should be representative of the entire cell area, not the centroid of the model cell.

Ground Water↗

Green fluorescent protein is lighting up fungal biology

Prasher ( 42 ) cloned a cDNA for the green fluorescent protein (GFP) gene from the jellyfish Aequorea victoria in 1992. Shortly thereafter, to the amazement of many investigators, this gene or derivatives thereof were successfully expressed and conferred fluorescence to bacteria and Caenorhabditis elegans cells in culture ( 10 , 31 ), followed by yeast ( 24 , 39 ), mammals ( 40 ), Drosophila ( 66 ), Dictyostelium ( 23 , 30 ), plants ( 28 , 49 ), and filamentous fungi ( 54 ). The tremendous success of GFP as a reporter can be attributed to unique qualities of this 238-amino-acid, 27-kDa protein which absorbs light at maxima of 395 and 475 nm and emits light at a maximum of 508 nm. The fluorescence of GFP requires only UV or blue light and oxygen, and therefore, unlike the case with other reporters (β-glucuronidase, β-galacturonidase, chloramphenicol acetyltransferase, and firefly luciferase) that rely on cofactors or substrates for activity, in vivo observation of gfp expression is possible with individual cells, with cell populations, or in whole organisms interacting with symbionts or environments in real time. Complications caused by destructive sampling, cell permeablization for substrates, or leakage of products do not occur. Furthermore, the GFP protein is extremely stable in vivo and has been fused to the C or N terminus of many cellular and extracellular proteins without a loss of activity, thereby permitting the tagging of proteins for gene regulation analysis, protein localization, or specific organelle labeling. The mature protein resists many proteases and is stable up to 65°C and at pH 5 to 11, in 1% sodium dodecyl sulfate or 6 M guanidinium chloride (reviewed in references 17 and 67 ), and in tissue fixed with formaldehyde, methanol, or glutaraldehyde. However, GFP loses fluorescence in methanol-acetic acid (3:1) and can be masked by autofluorescent aldehyde groups in tissue fixed with glutaraldehyde. Fluorescence is optimal at pH 7.2 to 8.0 ( 67 ). Limitations on GFP as a reporter for some applications are its low turnover rate, 2-h lag time for autoactivation of its chromophore, improper folding at high temperatures (37°C), which results in nonfluorescent and insoluble forms of the protein, and requirement for oxygen, which is not present in equal concentrations in all subcellular locations or cell types (reviewed in references 17 and 67 ). These characteristics of GFP, however, have not posed a problem for many applications, and mutant forms of GFP that have an ability to fold properly at high temperatures, increased solubility and fluorescence, reduced photobleaching ( 16 , 17 , 51 ), and reduced half-lives ( 1 ) have been developed. Coupled with fluorescence-activated cell sorting, confocal microscopy or quantitative image analysis techniques, GFP technology can be used to isolate transformed cells or specific cell types from populations of cells ( 14 ), to quantify gene expression of individual cells within whole organisms ( 8 ), or to assess the dispersal and biomass of organisms in complex environments, such as in animal or plant hosts ( 38 , 59 ), in biofilms ( 55 ), in fermentors ( 41 ), on leaf surfaces ( 53 , 61 ), or in soils ( 2 ). The vast majority of studies utilizing GFP expression in fungi have been with yeast (reviewed in reference 13 ). Ustilago maydis was the first filamentous fungus for which successful expression of gfp was reported ( 54 ), followed closely by Aspergillus nidulans ( 22 , 57 ) and Aureobasidium pullulans ( 61 ). Presently, gfp expression has been reported for 16 species comprising 12 genera of filamentous fungi, including Colletotrichum ( 21 , 44 ), Mycosphaerella ( 52 ), Magnaporthe ( 32 , 35 ), Cochliobolus ( 38 ), Trichoderma ( 2 , 70 ), Podospora ( 5 ), Sclerotinia ( 63 ), Schizophyllum ( 37 ), Aspergillus ( 20 , 47 , 50 ) and Phytophthora ( 7 , 62 ). In this review we draw on published reports, with the goal of providing an overview of GFP technology as it applies to the biology of filamentous fungi. These reports are not exhaustive of potential applications of GFP technology, as examples of genomic approaches to utilizing GFP in bacterial and yeast systems attest ( 4 , 46 , 60 , 65 ). Expression of gfp in filamentous fungi requires a gfp variant that is efficiently translated in fungi, a transformation system, and a fungal promoter that satisfies the requirements of a given experimental objective. Transformation of fungi has recently been reviewed by Gold et al. ( 26 ). Robinson and Sharon ( 44 ) suggest that GFP can actually be used to optimize transformation protocols. In addition to reporting the construction of a new fungal transformation vector that expresses SGFP under the control of the ToxA gene promoter from Pyrenophora tritici-repentis ( 12 ) and demonstrating its use in plant pathogens belonging to eight different genera of filamentous fungi ( Fusarium, Botrytis, Pyrenophora, Alternaria, Cochliobolus, Sclerotinia, Colletotrichum , and Verticillium ), in this review we also enumerate and describe a comprehensive list of vectors for expressing GFP in fungi.

Applied and Environmental Microbiology↗

Soft shell clams Mya arenaria with disseminated neoplasia demonstrate reverse transcriptase activity

Disseminated neoplasia (DN), a proliferative cell disorder of the circulatory system of bivalves, was first reported in oysters in 1969. Since that time, the disease has been determined to be transmissible through water-borne exposure, but the etiological agent has not been unequivocally identified. In order to determine if a viral agent, possibly a retrovirus, could be the causative agent of DN, transmission experiments were performed, using both a cell-free filtrate and a sucrose gradient-purified preparation of a cell-free filtrate of DN positive materials. Additionally, a PCR-enhanced reverse transcriptase assay was used to determine if reverse transcriptase was present in tissues or hemolymph from DN positive soft shell clams Mya arenaria. DN was transmitted to healthy clams by injection with whole DN cells, but not with cell-free flitrates prepared from either tissues from DN positive clams, or DN cells. The cell-free preparations from DN-positive tissues and hemolymph having high levels of DN cells in circulation exhibited positive reactions in the PCR-enhanced reverse transcriptase assay. Cell-free preparations of hemolymph from clams having low levels of DN (<0.1% of cells abnormal), hemocytes from normal soft shell clams, and normal soft shell clam tissues did not produce a positive reaction in the PCR enhanced reverse transcriptase assay.

Diseases of Aquatic Organisms↗

Metformin prevents endoplasmic reticulum stress-induced apoptosis through AMPK-PI3K-c-Jun NH2 pathway

Type 2 diabetes mellitus is thought to be partially associated with endoplasmic reticulum (ER) stress toxicity on pancreatic beta cells and the result of decreased insulin synthesis and secretion. In this study, we showed that a well-known insulin sensitizer, metformin, directly protects against dysfunction and death of ER stress-induced NIT-1 cells (a mouse pancreatic beta cell line) via AMP-activated protein kinase (AMPK) and phosphatidylinositol-3 (PI3) kinase activation. We also showed that exposure of NIT-1 cells to metformin (5mM) increases cellular resistance against ER stress-induced NIT-1 cell dysfunction and death. AMPK and PI3 kinase inhibitors abolished the effect of metformin on cell function and death. Metformin-mediated protective effects on ER stress-induced apoptosis were not a result of an unfolded protein response or the induced inhibitors of apoptotic proteins. In addition, we showed that exposure of ER stressed-induced NIT-1 cells to metformin decreases the phosphorylation of c-Jun NH(2) terminal kinase (JNK). These data suggest that metformin is an important determinant of ER stress-induced apoptosis in NIT-1 cells and may have implications for ER stress-mediated pancreatic beta cell destruction via regulation of the AMPK-PI3 kinase-JNK pathway.

Biochemical and Biophysical Research Communication↗

Effect of cryopreservation and in vitro culture of bovine fibroblasts on histone acetylation levels and in vitro development of hand-made cloned embryos

In this study, the relative acetylation levels of histone 3 in lysine 9 (H3K9ac) in cultured and cryopreserved bovine fibroblasts was measured and we determined the influence of the epigenetic status of three cultured (C1, C2 and C3) donor cell lines on the in vitro development of reconstructed bovine embryos. Results showed that cryopreservation did not alter the overall acetylation levels of H3K9 in bovine fibroblasts analysed immediately after thawing (frozen/thawed) compared with fibroblasts cultured for a period of time after thawing. However, reduced cleavage rates were noted in embryos reconstructed with fibroblasts used immediately after thawing. Cell passage affects the levels of H3K9ac in bovine fibroblasts, decreasing after P1 and donor cells with lower H3K9ac produced a greater frequency of embryo development to the blastocyst stage. Cryopreservation did not influence the total cell and ICM numbers, or the ICM/TPD ratios of reconstructed embryos. However, the genetic source of donor cells did influence the total number of cells and the trophectoderm cell numbers, and the cell passage influenced the total ICM cell numbers.

Zygote↗

Rewetting approximation for a simulator of flow in a surficial aquifer overlain by seasonally inundated wetlands

An approximation is presented to rectify situations occurring as part of the three-dimensional simulation of surficial aquifer flows in which surface grid cells become dry (zero saturated thickness) and then cannot readily receive flows that would again partially saturate them. The approximation permits the cells to be rewetted by adjusting dry cell pressures by the same amount as the pressure change computed for the uppermost nondry cells in the vertical column. This is continued in successive timesteps until the dry cells once again become partially or fully saturated. The rewetting procedure was tested in a recent transient simulation of flows in the surficial Biscayne aquifer of Dade County, Florida, which is partly covered by seasonally inundated wetlands. The uppermost layer of the model (layer 1) was used to represent overland sheetflow and grid cells of high equivalent hydraulic conductivity in this layer became dry and were rewetted seasonally, requiring a relatively robust representation of rewetting. The rewetting procedure made possible a simulation of overland sheetflow stages and aquifer water-table altitudes between 1945 and 1989 that was generally correct to within about 1 foot in inundated and noninundated regions. In a 6-year time period (1962–67) in which the behavior of the rewetting procedure was studied in detail, there were 4,723 simulated cell rewettings, of which 3,172 were in the overland sheetflow layer and 1,551 were in the uppermost aquifer layer (layer 2). The average head adjustment in a cell rewetting was 0.10 foot in layer 1 and 0.15 foot in layer 2. This study demonstrates an empirical approach to the problem of cell rewetting that might prove useful in other models of flow in surficial aquifers.

Groundwater↗

Isolation, growth, and metabolism of an obligately anaerobic, selenate- respiring bacterium, strain SES-3

A gram-negative, strictly anaerobic, motile vibrio was isolated from a selenate-respiring enrichment culture. The isolate, designated strain SES-3, grew by coupling the oxidation of lactate to acetate plus CO 2 with the concomitant reduction of selenate to selenite or of nitrate to ammonium. No growth was observed on sulfate or selenite, but cell suspensions readily reduced selenite to elemental selenium (Se 0 ). Hence, SES-3 can carry out a complete reduction of selenate to Se 0 . Washed cell suspensions of selenate-grown cells did not reduce nitrate, and nitrate-grown cells did not reduce selenate, indicating that these reductions are achieved by separate inducible enzyme systems. However, both nitrate-grown and selenate-grown cells have a constitutive ability to reduce selenite or nitrite. The oxidation of [ 14 C]lactate to 14 CO 2 coupled to the reduction of selenate or nitrate by cell suspensions was inhibited by CCCP (carbonyl cyanide m -chlorophenylhydrazone), cyanide, and azide. High concentrations of selenite (5 mM) were readily reduced to Se 0 by selenate-grown cells, but selenite appeared to block the synthesis of pyruvate dehydrogenase. Tracer experiments with [ 75 Se]selenite indicated that cell suspensions could achieve a rapid and quantitative reduction of selenite to Se 0 . This reduction was totally inhibited by sulfite, partially inhibited by selenate or nitrite, but unaffected by sulfate or nitrate. Cell suspensions could reduce thiosulfate, but not sulfite, to sulfide. These results suggest that reduction of selenite to Se 0 may proceed, in part, by some of the components of a dissimilatory system for sulfur oxyanions.

Nevada↗

Oxidation of methyl halides by the facultative methylotroph strain IMB-1

Washed cell suspensions of the facultative methylotroph strain IMB-1 grown on methyl bromide (MeBr) were able to consume methyl chloride (MeCl) and methyl iodide (MeI) as well as MeBr. Consumption of >100 μM MeBr by cells grown on glucose, acetate, or monomethylamine required induction. Induction was inhibited by chloramphenicol. However, cells had a constitutive ability to consume low concentrations (<20 nM) of MeBr. Glucose-grown cells were able to readily oxidize [ 14 C]formaldehyde to 14 CO 2 but had only a small capacity for oxidation of [ 14 C]methanol. Preincubation of cells with MeBr did not affect either activity, but MeBr-induced cells had a greater capacity for [ 14 C]MeBr oxidation than did cells without preincubation. Consumption of MeBr was inhibited by MeI, and MeCl consumption was inhibited by MeBr. No inhibition of MeBr consumption occurred with methyl fluoride, propyl iodide, dibromomethane, dichloromethane, or difluoromethane, and in addition cells did not oxidize any of these compounds. Cells displayed Michaelis-Menten kinetics for the various methyl halides, with apparent K s values of 190, 280, and 6,100 nM for MeBr, MeI, and MeCl, respectively. These results suggest the presence of a single oxidation enzyme system specific for methyl halides (other than methyl fluoride) which runs through formaldehyde to CO 2 . The ease of induction of methyl halide oxidation in strain IMB-1 should facilitate its mass culture for the purpose of reducing MeBr emissions to the atmosphere from fumigated soils.

Applied and Environmental Microbiology↗

Effects of ultraviolet-B radiation on fish: Histologic comparison of a UVB-sensitive and a UVB-tolerant species

Lahontan cutthroat trout Oncorhynchus clarki henshatvi were sensitive to simulated solar ultraviolet-B radiation (UVB) and exhibited grossly visible signs of sunburn upon exposure. Razorback suckers Xyrauchen texanus , however, were tolerant to simulated solar UVB and showed no grossly visible signs of exposure. Cutthroat trout also had considerably less of an unidentified, possibly photoprotective, substance in the skin than did razorback suckers. In an attempt to characterize the cellular response to simulated solar UVB exposure in the skin of these two species, we examined sections from UVB-exposed fish by light and electron microscopy. Cutthroat trout showed grossly visible signs of sunburn by 48 h. Histologic observations included a sloughing of the mucous cells, necrosis and edema in the epidermis and dermis, and, in some cases, secondary fungal infections. Razorback suckers did not show any visible signs of sunburn during 72 h of experimental exposure. Histologic analyses revealed that cell necrosis had occurred, but the severe necrosis and sloughing noted in cutthroat trout was not observed. An increase in epidermal thickness, apparently due to hypertrophy and hyperplasia of large PAS-negative cells, occurred in the razorback suckers. These cells contained a large central region of low electron density and appeared to be club cells. In some, extensive interdigitation of the electron-lucent cytoplasm with adjacent epithelial cell margins occurred. Near the surface of the epidermis these cells were larger and the interface with epithelial cells lacked complex interdigitation. These cells may contain the substance that appears to protect razorback suckers against UV-B radiation.

Journal of Aquatic Animal Health↗

Assessment of habitat use by juvenile Chinook salmon (Oncorhynchus tshawytscha) in the Willamette River Basin, 2020–21

We conducted a field study during 2020–21 to describe habitat use patterns of juvenile Chinook salmon ( Oncorhynchus tshawytscha ) in the mainstem Willamette, McKenzie, and Santiam Rivers and to evaluate how habitat suitability criteria affected the predictive accuracy of a hydraulic habitat model. Two approaches were used to collect habitat use data: a stratified sampling design was used to ensure that a representative sample of available habitats was included in our sampling; and a targeted sampling design was used to collect additional data in habitat cells where juvenile Chinook salmon were observed. Habitat attributes and fish presence data were collected in habitat cells that were approximately 2 square meters during April, June, and July. A total of 632 cells were sampled during the study and included habitat located in the main channel (373 cells), side channels (228 cells), and in alcoves (31 cells). Juvenile Chinook salmon were observed in 42 percent of the cells located in the main channel, 38 percent of the cells located in side channels, and 7 percent of the cells located in alcoves. We used logistic regression to develop resource selection functions for April, June, and July, which produced probability-based predictions of habitat use for juvenile Chinook salmon based on water velocity and water depth. The resource selection functions revealed a habitat shift by juvenile Chinook salmon to locations with higher water velocities and greater water depths from April to July as juvenile Chinook salmon size increased. The resource selection functions that we developed are an important addition to habitat modeling in the Willamette River basin because they were developed from in-basin data, capture seasonal differences in habitat use, and facilitate probability-based estimates of habitat use for juvenile Chinook salmon. These advancements will improve habitat modeling efforts for juvenile Chinook salmon during spring and summer months within the Willamette River.

Oregon↗

Nearshore sediment monitoring for the Stormwater Action Monitoring (SAM) Program, Puget Sound, western Washington

Chemicals such as metals and organics (polychlorinated biphenyl [PCBs], polybrominated diphenyl ethers [PBDEs], polycyclic aromatic hydrocarbons [PAHs], and phthalates) continue to enter Puget Sound, western Washington, from point sources (such as industrial and municipal outfalls) and combined sewer outfalls and non-point sources (such as stormwater runoff). Runoff during storm events has been identified as a major source of contamination entering Puget Sound and has been implicated in the degradation of nearshore habitats and biota. Metals, organic chemicals, and other pollutants are known to accumulate in sediments such as those present along the shoreline of Puget Sound. In addition to chemical contaminants, small plastic particles (known as microplastics), found in marine waters of Puget Sound and suspected of being in aquatic sediments, are a potential concern because they can be ingested by animals and are suspected of transporting sorbed chemicals such as PCBs and metals. The Stormwater Work Group of Puget Sound (SWG) (composed of State and municipal stormwater permittees, and other stakeholders) developed a strategy to address sediment conditions in the nearshore environment of Puget Sound. As part of this strategy, the SWG developed a regional stormwater monitoring strategy designed to inform monitoring requirements in National Pollutant Discharge Elimination System (NPDES) stormwater permits issued by the Washington State Department of Ecology (Ecology). The monitoring program is referred to as the Stormwater Action Monitoring (SAM). The overall focus of the work described in this report is to address one of the goals of SAM, which is to characterize the status, spatial extent, and quality of Puget Sound sediment chemicals in the nearshore urban areas. The nearshore urban areas are defined as areas parallel to established Urban Growth Areas (UGAs) using a spatially balanced probabilistic Generalized Random Tessellation Stratified (GRTS) sampling design. One of the benefits of the GRTS sampling design used for this study is that it allows one to efficiently extrapolate from a relatively small number of sampled nearshore sites to the entire nearshore shoreline within the 2011 defined UGA boundaries of Puget Sound. In addition to characterizing nearshore sediment chemical concentrations, this study also characterized the abundance of microplastics in the nearshore sediment. A total of 41 randomly selected sites were sampled throughout Puget Sound in summer and early autumn of 2016. All sampling sites were located at 6 feet below the Mean Lower Low Water line. The top 2–3 centimeters of sediment were collected using a boat-mounted, pre-cleaned stainless-steel box corer. All chemical samples were sieved to 2 millimeters and placed in appropriate containers for chemical analysis for PCBs, PBDEs, PAHs, phthalates, metals, total organic carbon, and grain size. Pre-sieved sediment samples were stored in glass containers for microplastic analysis. Nearshore sediment chemical concentrations were summarized using numerous statistical approaches to examine the minimum, mean, and maximum concentrations for each of the compounds analyzed and to compare the results to criteria and other nearshore and marine sediment studies. The GRTS sampling design also allowed the authors to assess the percentage of the UGA nearshore environment that did not meet established standards or criteria for each chemical analyzed. Additionally, regression and machine learning statistical analyses were used to examine relations between measured chemical concentrations, and land cover and geologic features at multiple scales within the watersheds adjacent to sampling sites. The influence of marine hydrodynamic factors on nearshore sediment chemical concentrations was statistically evaluated with nonparametric methods by assigning each sampling site to one of five nearshore drift cell types based on its location. The Puget Sound shoreline can be divided into segments, referred to as drift cells, based on the movement of sediment along the shore by waves. Each drift cell type has a unique influence on nearshore sediment transport. The nearshore sediment chemical concentrations for organics and metals generally were low, and in most cases less than Washington State criteria. The concentrations of some PAHs were greater than the criteria, but these exceedances were limited to one or two sites. The results of the probabilistic study design determined that, for the PAHs examined, 96 percent or more of the 1,344 km of shoreline represented by this study had concentrations less than any established criteria. For the remaining organics (PCBs and PBDEs), the probabilistic study design indicates that more than 98 percent of shoreline examined had concentrations less than criteria or proposed standards. For the metals, the results of the study indicate that 100 percent of the nearshore sediment had concentrations less than the criteria. The relations between sediment organic and metal concentrations, and adjacent watershed land cover and the particle size of the samples, were determined to be weakly related. Although weakly related, the particle size of the sediment in a sample typically explained more of the variation in metal concentrations than organics. While the measured watershed attributes adjacent to the sampling sites and sediment size of the samples were weakly related to chemical concentrations, they were significantly related to unique drift cells along the shoreline of Puget Sound known as drift cells. Each drift cell represents a long-term directional transport of sediment from its source to its depositional zone. Sediment chemical concentrations were significantly higher in drift cells with limited sediment movement compared to those with higher sediment transport energy. Microplastics in the nearshore sediment ranged from 0.02 to 0.65 pieces per gram of sediment, with a mean of 0.19 pieces per gram of sediment, and were dominated by small fibers (355–1,000 micrometers). Like chemical concentrations, microplastics concentrations in the nearshore sediment were poorly related to watershed land cover. Although not significantly different, microplastics concentrations generally were higher in the low energy drift cells compared to the high energy drift cells. The results of this study provide a statistically valid status assessment of current nearshore sediment chemical conditions throughout Puget Sound in those areas adjacent to defined UGAs. In addition to the study findings of relatively low concentrations of PCBs, PBDEs, PAHs, phthalates, and metals, the study design provides a statistically valid tool for evaluating changes in these compounds over time if future nearshore sediment assessments are done. Furthermore, the assessment of microplastic abundance represents the first study of its kind that can be used as a benchmark for future evaluations. The results of this study will help inform Ecology in the implementation of monitoring requirements as part of its NPDES stormwater permitting process.

Washington↗

Distribution of yellow crazy ants (Anoplolepis gracilipes) and notes on other ant species at Wake Atoll, 6-21 October, 2023

Wake Atoll, located in the central Pacific Ocean, is a tropical coral atoll comprised of three islands totaling 696 ha of emergent land. Wake Atoll supports at least 17 species of non-native ants, but one species, the yellow crazy ant ( Anoplolepis gracilipes ), is especially harmful to ecosystems, including seabirds, which are federally protected species and nest throughout Wake Atoll. Our objective was to map the distribution of yellow crazy ants (YCA) across Wake Atoll to provide U.S. Air Force managers with information that could inform an eradication strategy for this species. In addition, we documented the presence of other ant species identified during our survey. We used a 50 x 50-m cell-based sampling grid to survey for YCA and other ant species. YCA were widespread across the atoll, occupying all three islands and were detected in 566 of the 3,675 (15.4%) cells surveyed, covering an area of about 142 ha. The highest proportion of cells containing YCA was on Wilkes Island (99 of 400 cells or 24.8%), followed by Wake Island (395 of 2,760 cells or 14.3%) and Peale Island (72 of 518 cells or 14.0%). YCA detections on Wilkes Island were restricted to Wilkes South (99 of 202 cells or 49.0%); no YCA were found on Wilkes North, an important area that supports the most diverse assemblage of nesting seabirds on Wake Atoll. Based on technologies developed to eradicate YCA from Johnston Atoll and elsewhere, eradicating YCA from Wake Atoll appears technically feasible. The occurrence of discrete populations (i.e., clusters of occupied cells separated from other clusters of occupied cells) within and among islands could allow a stepwise strategy where the short-term outcome of the treatment of one population is independent of the outcome of another treatment. Without action, the high-density YCA population on Wilkes South could breach the narrow (~50 m), sometimes dry, channel separating Wilkes North and South, posing a serious threat to nesting seabirds present on Wilkes North. Eradicating YCA from Wilkes South would create a buffer from YCA and reduce the threat of northward movement of YCA. We also detected 12 ant species other than YCA during our survey, including Lepisiota frauenfeldi , a new record for the atoll.

Hawaii Cooperative Studies Unit Technical Report↗

Toxic effects of dietary methylmercury on immune system development in nestling American kestrels ( Falco sparverius )

This study evaluated the effects of dietary methylmercury (MeHg) on immune system development in captive-reared nestling American kestrels ( Falco sparverius ) to determine whether T cell&ndash;mediated and antibody-mediated adaptive immunity are targets for MeHg toxicity at environmentally relevant concentrations. Nestlings received various diets, including 0 (control), 0.6, and 3.9 &mu;g/g (dry wt) MeHg for up to 18 d posthatch. Immunotoxicity endpoints included cell-mediated immunity (CMI) using the phytohemagglutinin (PHA) skin-swelling assay and antibody-mediated immune response via the sheep red blood cell (SRBC) hemagglutination assay. T cell&ndash; and B cell&ndash;dependent histological parameters in the spleen, thymus, and bursa of Fabricius were correlated with the functional assays. For nestlings in the 0.6 and 3.9 &mu;g/g MeHg groups, CMI was suppressed by 73 and 62%, respectively, at 11 d of age. Results of this functional assay were correlated with T cell&ndash;dependent components of the spleen and thymus. Dose-dependent lymphoid depletion in spleen tissue directly affected the proliferation of T-lymphocyte populations, insofar as lower stimulation indexes from the PHA assay occurred in nestlings with lower proportions of splenic white pulp and higher THg concentrations. Nestlings in the 3.9 &mu;g/g group also exhibited lymphoid depletion and a lack of macrophage activity in the thymus. Methylmercury did not have a noticeable effect on antibody-mediated immune function or B cell&ndash;dependent histological correlates. We conclude that T cell&ndash;mediated immunosuppression is the primary target of MeHg toward adaptive immunity in developing kestrels. This study provides evidence that environmentally relevant concentrations of MeHg may compromise immunocompetence in a developing terrestrial predator and raises concern regarding the long-term health effects of kestrels that were exposed to dietary MeHg during early avian development.

Environmental Toxicology and Chemistry↗

The memory of the accreting plate boundary and the continuity of fracture zones

A detailed aeromagnetic anomaly map of the Mesozoic seafloor-spreading lineations southwest of Bermuda reveals the dominant magnetic grain of the oceanic crust and the character of the accreting boundary at the time of crustal formation. The magnetic anomaly pattern is that of a series of elongate lobes perpendicular to the fracture zone (flowline) trends. The linear sets of magnetic anomaly peaks and troughs have narrow regions of reduced amplitude anomalies associated with the fracture zones. During the period of Mesozoic geomagnetic polarity reversals (when 1200 km of central North Atlantic seafloor formed), the Atlantic accreting boundary consisted of stationary, elongate, spreading center cells that maintained their independence even though sometimes only minor spatial offsets existed between cells. Normal oceanic crustal structure was formed in the spreading center cells, but structural anomalies and discontinuities characteristic of fracture zones were formed at their boundaries, which parallel flowlines of Mesozoic relative plate motion in the central North Atlantic. We suggest that the memory for a stationary pattern of independent spreading center cells resides in the young brittle lithosphere at the accreting boundary where the lithosphere is weakest; here, each spreading center cell independently goes through its cylce of stress buildup, stress release, and crustal accretion, after which its memory is refreshed. The temporal offset between the peaks of the accretionary activity that takes place within each cell may provide the mechanism for maintaining the independence of adjacent spreading center cells through times when no spatial offset between the cells exists.

Earth and Planetary Science Letters↗

Flow cytometric analysis of crayfish haemocytes activated by lipopolysaccharides

Lipopolysaccharides (LPS) from Gram-negative bacteria are strong stimulators of white river crayfish, Procambarus zonangulus, haemocytes in vitro. Following haemocyte treatment with LPS and with LPS from rough mutant R5 (LPS Rc) from Salmonella minnesota, flow cytometric analysis revealed a conspicuous and reproducible decrease in cell size as compared to control haemocytes. These LPS molecules also caused a reduction in haemocyte viability as assessed by flow cytometry with the fluorescent dyes calcein-AM and ethidium homodimer. The onset of cell size reduction was gradual and occurred prior to cell death. Haemocytes treated with LPS from S. minnesota without the Lipid A moiety (detoxified LPS) decreased in size without a reduction of viability. The action of LPS on crayfish haemocytes appeared to be related to the activation of the prophenoloxidase system because phenoloxidase (PO)-specific activity in the supernatants from control and detoxified LPS-treated cells was significantly lower than that from LPS and LPS-Rc treated cells (P < 0.05). Furthermore, addition of trypsin inhibitor to the LPS treatments caused noticeable delays in cell size and viability changes. These patterns of cellular activation by LPS formulations indicated that crayfish haemocytes react differently to the polysaccharide and lipid A moieties of LPS, where lipid A is cytotoxic and the polysaccharide portion is stimulatory. These effects concur with the general pattern of mammalian cell activation by LPS, thereby indicting commone innate immune recognition mechanisms to bacterial antigens between cells from mammals and invertebrates. These definitive molecular approaches used to verify and identify mechanisms of invertbrate haemocyte responses to LPS could be applied with other glycoconjugates, soluble mediators, or xenobiotic compounds.

Fish and Shellfish Immunology↗

Hot, wet and rare: Modelling the occupancy dynamics of the narrowly distributed Dixie Valley toad

Context: Small population sizes and no possibility of metapopulation rescue put narrowly distributed endemic species under elevated risk of extinction from anthropogenic change. Desert spring wetlands host many endemic species that require aquatic habitat and are isolated by the surrounding xeric terrestrial habitat. Aims: We sought to model the occupancy dynamics of the Dixie Valley toad ( Anaxyrus williamsi ), a recently described species endemic to a small desert spring wetland complex in Nevada, USA. Methods: We divided the species’ range into 20 m × 20 m cells and surveyed for Dixie Valley toads at 60 cells during six primary periods from 2018 to 2021, following an occupancy study design. We analysed our survey data by using a multi-state dynamic occupancy model to estimate the probability of adult occurrence, colonisation, site survival, and larval occurrence and the relationship of each to environmental covariates. Key results: The detection probabilities of adult and larval toads were affected by survey length and time of day. Adult Dixie Valley toads were widely distributed, with detections in 75% of surveyed cells at some point during the 3-year study, whereas larvae were observed only in 20% of cells during the study. Dixie Valley toad larvae were more likely to occur in cells far from spring heads with a high coverage of surface water, low emergent vegetation cover, and water temperatures between 20°C and 28°C. Adult toads were more likely to occur in cells with a greater coverage of surface water and water depth >10 cm. Cells with more emergent vegetation cover and surface water were more likely to be colonised by adult toads. Conclusions: Our results showed that Dixie Valley toads are highly dependent on surface water in both spring and autumn. Adults and larvae require different environmental conditions, with larvae occurring farther from spring heads and in fewer cells. Implications: Disturbances to the hydrology of the desert spring wetlands in Dixie Valley could threaten the persistence of this narrowly distributed toad.

Wildlife Research↗

A regional model comparison between MODPATH and MT3D of groundwater travel time distributions

Groundwater quality changes in wells and streams lag behind changes to land use due to groundwater travel times. Two contaminant transport methods were compared to assess differences in their simulated travel time distributions (TTDs) to streams and wells in the Wisconsin Central Sands. MODPATH simulates advective groundwater flow with particle tracking, while MT3D simulates age-mass using a finite difference solution without dispersion to allow for direct comparison of the two methods. MODPATH appropriately simulates groundwater TTDs from the water table to surface discharge but is subject to inaccuracies at weak-sink well cells due to the flow-model grid discretization and imprecise location of well discharge within well cells. MT3D better represents weak-sink well cells since it removes mass in proportion to the prescribed pumping rate, although travel time within well cells is neglected. Conversely, MT3D's treatment of surface water boundary cells is not as accurate as MODPATH because mass should be removed from the water table rather than the full cell volume. MT3D simulations of TTDs can also be confounded by the instantaneous vertical distribution of mass introduced throughout recharge cells instead of at the water table, which initiates mass along deeper flow paths. We evaluated 9 MODPATH and 13 MT3D implementations, generating differences in median travel times of up to 18 years. Both methods have strengths and weaknesses, with MT3D better representing weak-sink well cell behavior and MODPATH better representing surficial recharge and discharge. The effect of these characteristics on simulated TTDs, along with ideas for ameliorating method weaknesses, is discussed.

Wisconsin↗