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Adaptive radiations are generally thought to occur soon after a lineage invades a region offering high levels of ecological opportunity. However, few adaptive radiations beyond a handful of exceptional examples are known, so a comprehensive understanding of their dynamics is still lacking. Here, we present a novel case of an island species flock of freshwater fishes with a radically different tempo of adaptive history than that found in many popular evolutionary model systems. Using a phylogenomic data set combined with simultaneous Bayesian estimation of divergence times and trait-based speciation and extinction models, we show that the New Zealand Gobiomorphus gudgeons comprise a monophyletic assemblage, but surprisingly, the radiation did not fully occupy freshwater habitats and explosively speciate until more than 10 myr after the lineage invaded the islands. This shift in speciation rate was not accompanied by an acceleration in the rate of morphological evolution in the freshwater crown clade relative to the other species, but is correlated with a reduction in head pores and scales as well as an increase in egg size. Our results challenge the notion that clades always rapidly exploit ecological opportunities in the absence of competing lineages. Instead, we demonstrate that adaptive radiation can experience a slow start before undergoing accelerated diversification and that lineage and phenotypic diversification may be uncoupled in young radiations. [Adaptive radiation; Eleotridae; freshwater; Gobiomorphus; New Zealand.]

Oil and gas development and transportation in many areas of the world, such as the oil sands region of Alberta, Canada, are heavily monitored to minimize the environmental impacts of development and the risk of oil spills. However, oil spills to aquatic environments still occur. Although the science of oil spills has received considerable attention of late, uncertainty still remains in evaluating the fate and transport of oil spills as well as the effects of those spills on aquatic biota. Experiments using meso-scale spill tanks were used to examine the physical and chemical behavior of two types of oil, conventional crude (CC) and diluted bitumen (DB), under similar environmentally relevant scenarios (i.e., volume of spill, temperature, duration, wave action, and presence of river sediment). In addition, biological impact assessments via sediment toxicity testing collected from the oil spill tests were conducted. Sediments were evaluated for acute toxicity using three standard sediment test species: Hyalella azteca, Lumbriculus variegatus, and Chironomus dilutus. Sediments collected from the CC simulated spill showed a higher level of acute toxicity than sediments collected from spills with DB. Higher toxicity in the CC-contaminated sediment was supported by higher concentrations of low molecular weight polycyclic aromatic hydrocarbons (PAHs) when compared with the DB-contaminated sediment, while the remaining PAH profile was similar between the contaminated sediments. The use of a meso-scale spill tank in combination with sediment bioassays allowed for the evaluation of oil spills under controlled and environmentally relevant conditions (e.g., nearshore high sediment loading river), and in turn provides assessors with additional information to identify the appropriate mitigation and remediation efforts needed in the event of future spills. Environ Toxicol Chem 2022;41:2797-2807. © 2022 SETAC.

Evolutionary transitions between marine and freshwater ecosystems have occurred repeatedly throughout the phylogenetic history of fishes. The theory of ecological opportunity predicts that lineages that colonize species-poor regions will have greater potential for phenotypic diversification than lineages invading species-rich regions. Thus, transitions between marine and freshwaters may promote phenotypic diversification in trans-marine/freshwater fish clades. We used phylogenetic comparative methods to analyze body size data in nine major fish clades that have crossed the marine/freshwater boundary. We explored how habitat transitions, ecological opportunity, and community interactions influenced patterns of phenotypic diversity. Our analyses indicated that transitions between marine and freshwater habitats did not drive body size evolution, and there are few differences in body size between marine and freshwater lineages. We found that body size disparity in freshwater lineages is not correlated with the number of independent transitions to freshwaters. We found a positive correlation between body size disparity and overall species richness of a given area, and a negative correlation between body size disparity and diversity of closely related species. Our results indicate that the diversity of incumbent freshwater species does not restrict phenotypic diversification, but the diversity of closely related taxa can limit body size diversification. Ecological opportunity arising from colonization of novel habitats does not seem to have a major effect in the trajectory of body size evolution in trans-marine/freshwater clades. Moreover, competition with closely related taxa in freshwaters has a greater effect than competition with distantly related incumbent species.

Background Carp gudgeons (genus Hypseleotris) are a prominent part of the Australian freshwater fish fauna, with species distributed around the western, northern, and eastern reaches of the continent. We infer a calibrated phylogeny of the genus based on nuclear ultraconserved element (UCE) sequences and using Bayesian estimation of divergence times, and use this phylogeny to investigate geographic patterns of diversification with GeoSSE. The southeastern species have hybridized to form hemiclonal lineages, and we also resolve relationships of hemiclones and compare their phylogenetic placement in the UCE phylogeny with a hypothesis based on complete mitochondrial genomes. We then use phased SNPs extracted from the UCE sequences for population structure analysis among the southeastern species and hemiclones. Results Hypseleotris cyprinoides, a widespread euryhaline species known from throughout the Indo-Pacific, is resolved outside the remainder of the species. Two Australian radiations comprise the bulk of Hypseleotris, one primarily in the northwestern coastal rivers and a second inhabiting the southeastern region including the Murray–Darling, Bulloo-Bancannia and Lake Eyre basins, plus coastal rivers east of the Great Dividing Range. Our phylogenetic results reveal cytonuclear discordance between the UCE and mitochondrial hypotheses, place hemiclone hybrids among their parental taxa, and indicate that the genus Kimberleyeleotris is nested within the northwestern Hypseleotris radiation along with three undescribed species. We infer a crown age for Hypseleotris of 17.3 Ma, date the radiation of Australian species at roughly 10.1 Ma, and recover the crown ages of the northwestern (excluding H. compressa) and southeastern radiations at 5.9 and 7.2 Ma, respectively. Range-dependent diversification analyses using GeoSSE indicate that speciation and extinction rates have been steady between the northwestern and southeastern Australian radiations and between smaller radiations of species in the Kimberley region and the Arnhem Plateau. Analysis of phased SNPs confirms inheritance patterns and reveals high levels of heterozygosity among the hemiclones. Conclusions The northwestern species have restricted ranges and likely speciated in allopatry, while the southeastern species are known from much larger areas, consistent with peripatric speciation or allopatric speciation followed by secondary contact. Species in the northwestern Kimberley region differ in shape from those in the southeast, with the Kimberley species notably more elongate and slender than the stocky southeastern species, likely due to the different topographies and flow regimes of the rivers they inhabit.

To describe the technique of combined center of rotation of angulation (CORA)-based leveling osteotomy (CBLO) with tibial tuberosity transposition (TTT) and to compare the load to failure between CBLO combined with TTT and CBLO or TTT alone.Ex vivo study.Twelve pairs of cadaveric pelvic limbs.Six pairs of cadaveric tibia were tested in each group (CBLO-TTT versus CBLO) and (CBLO-TTT versus TTT) with each limb randomly assigned to a treatment group. Construct stability was determined by applying a tensile force to each patellar tendon until failure occurred. Load at failure and mode of failure were recorded for each specimen.No difference in mean load to failure was identified between CBLO-TTT (897 N) and CBLO alone (943 N) (P = .81). There was also no difference in the mean load to failure between the CBLO-TTT (928 N) and TTT alone (1046 N) (P = .12).Performing a TTT in combination with a CBLO does not weaken the construct failure to load when compared with each procedure performed alone.A combined CBLO and TTT could be considered a viable option for concurrent management of a cranial cruciate ligament deficient stifle and medial patella luxation.

International audience; To camouflage themselves on the seafloor, European cuttlefish Sepia officinalis control the expression of about 30 pattern components to produce a range of body patterns.(1) If each component were under independent control, cuttlefish could produce at least 2(30) patterns. To examine how cuttlefish deploy this vast potential, we recorded cuttlefish on seven experimental backgrounds, each designed to resemble a pattern component, and then compared their responses to predictions of two models of sensory control of component expression. The body pattern model proposes that cuttlefish integrate low-level sensory cues to categorize the background and co-ordinate component expression to produce a small number of overall body patterns.(2-4) The feature matching model proposes that each component is expressed in response to one (or more) local visual features, and the overall pattern depends upon the combination of features in the background. Consistent with the feature matching model, six of the backgrounds elicited a specific set of one to four components, whereas the seventh elicited eleven components typical of a disruptive body pattern. This evidence suggests that both modes of control are important, and we suggest how they can be implemented by a recent hierarchical model of the cuttlefish motor system.(5)(,)(6).

In the present study, we compared the toxicity and associated chemical characterizations of flowback and produced water (FPW) collected from a single horizontal hydraulically fractured well at different time points during FPW production. Since few studies on whole mixture toxicity related to FPW exist, our aims were to determine both overall toxicity of the FPW mixture in a suite of organisms (Daphnia magna, Lumbriculus variegatus, Danio rerio, and Oncorhynchus mykiss) and also determine if toxicity changes depending on variation in FPW chemical properties as a function of time sampled (1.33, 72, and 228 h FPW samples collected immediately post-well production onset were analyzed in current study). FPW chemical composition was determined via quadra-pole inductively coupled plasma – mass spectrometry/mass spectrometry (ICP-MS/MS), full-scan high performance liquid chromatography/Orbitrap mass spectrometry (HPLC/Orbitrap-MS), and gas chromatography-mass spectrometry (GC-MS). We observed that FPW sampled later in the production process contained higher ion and total dissolved solids concentrations, whereas the highest concentrations of dissolved organic compounds were observed in the earliest FPW sample analyzed. Toxicity associated with FPW exposure was deemed to be species-specific to a certain extent, but general trends revealed the earliest FPW sampled contained highest toxic potential. Accordingly, we theorize that although the saline conditions of FPW are the foremost toxicological drivers to freshwater organisms, dissolved organics associated with FPW significantly contribute to the overall toxicity of exposed organisms.

Spiny-rayed fishes (Acanthomorpha) dominate modern marine habitats and account for more than a quarter of all living vertebrate species. Previous time-calibrated phylogenies and patterns from the fossil record explain this dominance by correlating the origin of major acanthomorph lineages with the Cretaceous-Palaeogene mass extinction. Here we infer a time-calibrated phylogeny using ultraconserved elements that samples 91.4% of all acanthomorph families and investigate patterns of body shape disparity. Our results show that acanthomorph lineages steadily accumulated throughout the Cenozoic and underwent a significant expansion of among-clade morphological disparity several million years after the end-Cretaceous. These acanthomorph lineages radiated into and diversified within distinct regions of morphospace that characterize iconic lineages, including fast-swimming open-ocean predators, laterally compressed reef fishes, bottom-dwelling flatfishes, seahorses and pufferfishes. The evolutionary success of spiny-rayed fishes is the culmination of multiple species-rich and phenotypically disparate lineages independently diversifying across the globe under a wide range of ecological conditions.

Hydraulic fracturing creates large volumes of flowback and produced water (FPW). The waste is a complex mixture of organic and inorganic constituents. Although the acute toxicity of FPW to freshwater organisms has been studied, few have attempted to discern the interaction between organic and inorganic constituents within this matrix and its role in toxicity. In the present study, bioaccumulation assays (7-d uptake and 7-d elimination period) with FPW (1% dilution) were conducted with the freshwater oligochaete, Lumbriculus variegatus, to evaluate the toxicokinetics of inorganic elements. To evaluate the interacting role of organics, bioaccumulation of elements in unmodified FPW was compared to activated carbon treated FPW (AC-modified). Differences in uptake and elimination rates as well as elimination steady state concentrations between unmodified and AC-modified treatments indicated that the organics play an important role in the uptake and depuration of inorganic elements in FPW. These differences in toxicokinetics between treatments aligned with observed growth rates in the worms which were higher in the AC-modified treatment. Whether growth differences resulted from increased accumulation and changes in toxicokinetic rates of inorganics or caused by direct toxicity from the organic fraction of FPW itself is still unknown and requires further research.

Influence of sediment particle size on the desorption, bioavailability, and bioaccumulation potential of cypermethrin was investigated in the present study using two biomimetic techniques (Tenax extraction and solid-phase microextraction (SPME)) and bioaccumulation testing with Lumbriculus variegatus. A field-collected sediment was wet sieved to obtain five particle-size fractions (20, 20-63, 63-180, 180-500, and500 μm) and used for cypermethrin spiking. The finest sediment (20 μm) had the highest rapid desorption fraction (F

The push to make bioassays more sensitive has meant an increased duration of testing to look at more chronic endpoints. To conduct these longer bioassays through the use of traditional bioassay methods can be difficult, as many traditional bioassays have employed manual water changes, which take considerable time and effort. To that end, static-renewal systems were designed to provide researchers a technique to ease the manual water change burden. One of the most well-known static-renewal designs, the static intermittent renewal system (STIR) was produced by the United States Environmental Protection Agency in 1993. This system is still being used in laboratories across the globe today. However, these initial designs have become rather dated as new technologies and methods have been developed that make these systems easier to build and operate. The following information details changes to the initial design and a proof of concept experiment with the benthic invertebrate, Chironomus tepperi, to validate the modifications to the original system.

Hydraulic fracturing of horizontal wells is a cost effective means for extracting oil and gas from low permeability formations. Hydraulic fracturing often produces considerable volumes of flowback and produced water (FPW). FPW associated with hydraulic fracturing has been shown to be a complex, often brackish mixture containing a variety of anthropogenic and geogenic compounds. In the present study, the risk of FPW releases to aquatic systems was studied using the model benthic invertebrate, Lumbriculus variegatus and field-collected FPW from a fractured well in Alberta. Acute, chronic, and pulse toxicity were evaluated to better understand the implications of accidental FPW releases to aquatic environments. Although L. variegatus is thought to have a high tolerance to many stressors, acute toxicity was significant at low concentrations (i.e. high dilutions) of FPW (48 h LC50: 4–5%). Chronic toxicity (28 d)of FPW in this species was even more pronounced with LC50s (survival/reproduction) and EC50s (total mass) at dilutions as low as 0.22% FPW. Investigations evaluating pulse toxicity (6 h and 48 h exposure) showed a significant amount of latent mortality occurring when compared to the acute results. Additionally, causality in acute and chronic bioassays differed as acute toxicity appeared to be primarily driven by salinity, which was not the case for chronic toxicity, as other stressors appear to be important as well. The findings of this study show the importance of evaluating multiple exposure regimes, the complexity of FPW, and also shows the potential aquatic risk posed by FPW releases. Hydraulic fracturing waste showed significant toxicological effects to Lumbriculus variegatus under acute, pulse, and chronic exposures.

The discovery of haloperidol catalyzed a breakthrough in our understanding of the biochemical basis of schizophrenia, improved the treatment of psychosis, and facilitated deinstitutionalization. In doing so, it solidified the role for chemical neuroscience as a means to elucidate the molecular underpinnings of complex neuropsychiatric disorders. In this Review, we will cover aspects of haloperidol's synthesis, manufacturing, metabolism, pharmacology, approved and off-label indications, and adverse effects. We will also convey the fascinating history of this classic molecule and the influence that it has had on the evolution of neuropsychopharmacology and neuroscience.

Gobies, sleepers, and cardinalfishes represent major clades of a species rich radiation of small bodied, ecologically diverse percomorphs (Gobiaria). Molecular phylogenetics has been crucial to resolving broad relationships of sleepers and gobies (Gobioidei), but the phylogenetic placements of cardinalfishes and nurseryfishes, as reciprocal or sequential sister clades to Gobioidei, are uncertain. In order to evaluate relationships among and within families we used a phylogenetic data mining approach to generate densely sampled trees inclusive of all higher taxa. We utilized conspecific amino acid homology to improve alignment accuracy, included ambiguously identified taxa to increase taxon sampling density, and resampled individual gene alignments to filter rogue sequences before concatenation. This approach yielded the most comprehensive tree yet of Gobiaria, inferred from a sparse (17 percent-complete) supermatrix of one ribosomal and 22 protein coding loci (18,065 characters), comprised of 50 outgroup and 777 ingroup taxa, representing 32 percent of species and 68 percent of genera. Our analyses confirmed the lineage-based classification of gobies with strong support, identified sleeper clades with unforeseen levels of systematic uncertainty, and quantified competing phylogenetic signals that confound resolution of the root topology. We also discovered that multilocus data completeness was related to maximum likelihood branch support, and verified that the phylogenetic uncertainty of shallow relationships observed within goby lineages could largely be explained by supermatrix sparseness. These results demonstrate the potential and limits of publicly available sequence data for producing densely-sampled phylogenetic trees of exceptionally biodiverse groups.

Agricultural use of neonicotinoid insecticides is increasing worldwide, posing a risk to nontarget organisms. The present study investigated developmental toxicity of a widely used neonicotinoid, acetamiprid, to zebrafish embryos. Sublethal (malformations, hatchability, heart rate, body length, alteration of spontaneous movement and touch responses) and lethal effects were monitored during exposure period from 6 h post fertilization (hpf) to 120 hpf. Zebrafish embryos exhibited significant mortality and teratogenic effects at acetamiprid concentration greater than 263 mg/L, with bent spine being the main malformation. Toxicity spectra were constructed to rank the sensitivity of individual end points to acetamiprid exposure and impaired spontaneous movement was the most sensitive end point of those tested. The present study provides the basis for understanding developmental toxicity of acetamiprid exposure to zebrafish embryos. This information is critical for future studies evaluating aquatic risk from neonicotinoids as little is known regarding adverse effects of neonicotinoids to aquatic vertebrate species.

Through epigenetic and other regulatory functions, the histone deacetylase (HDAC) family of enzymes has emerged as a promising therapeutic target for central nervous system and other disorders. Here we report on the synthesis and functional characterization of new HDAC inhibitors based structurally on tianeptine, a drug used primarily to treat major depressive disorder (MDD) that has a poorly understood mechanism of action. Since the chemical structure of tianeptine resembles certain HDAC inhibitors, we profiled the in vitro HDAC inhibitory activity of tianeptine and demonstrated its ability to inhibit the lysine deacetylase activity of a subset of class I HDACs. Consistent with a model of active site Zn2+ chelation by the carboxylic acid present in tianeptine, newly synthesized analogues containing either a hydroxamic acid or ortho-aminoanilide exhibited increased potency and selectivity among the HDAC family. This in vitro potency translated to improved efficacy in a panel of high-content imaging assays designed to assess HDAC target engagement and functional effects on critical pathways involved in neuroplasticity in both primary mouse neurons and, for the first time, human neurons differentiated from pluripotent stem cells. Most notably, tianeptinaline, a class I HDAC-selective analogue of tianeptine, but not tianeptine itself, increased histone acetylation, and enhanced CREB-mediated transcription and the expression of Arc (activity-regulated cytoskeleton-associated protein). Systemic in vivo administration of tianeptinaline to mice confirmed its brain penetration and was found to enhance contextual fear conditioning, a behavioral test of hippocampal-dependent memory. Tianeptinaline and its derivatives provide new pharmacological tools to dissect chromatin-mediated neuroplasticity underlying memory and other epigenetically related processes implicated in health and disease.

Extinction and colonization dynamics are critical to understanding the evolution and conservation of metapopulations. However, traditional field studies of extinction-colonization are potentially fraught with detection bias and have rarely been validated. Here, we provide a comparison of molecular and field-based approaches for assessment of the extinction-colonization dynamics of tidewater goby (Eucyclogobius newberryi) in northern California. Our analysis of temporal genetic variation across 14 northern California tidewater goby populations failed to recover genetic change expected with extinction-colonization cycles. Similarly, analysis of site occupancy data from field studies (94 sites) indicated that extinction and colonization are very infrequent for our study populations. Comparison of the approaches indicated field data were subject to imperfect detection, and falsely implied extinction-colonization cycles in several instances. For northern California populations of tidewater goby, we interpret the strong genetic differentiation between populations and high degree of within-site temporal stability as consistent with a model of drift in the absence of migration, at least over the past 20-30 years. Our findings show that tidewater goby exhibit different population structures across their geographic range (extinction-colonization dynamics in the south vs. drift in isolation in the north). For northern populations, natural dispersal is too infrequent to be considered a viable approach for recolonizing extirpated populations, suggesting that species recovery will likely depend on artificial translocation in this region. More broadly, this work illustrates that temporal genetic analysis can be used in combination with field data to strengthen inference of extinction-colonization dynamics or as a stand-alone tool when field data are lacking.

Recent studies indicate a role for the constitutive androstane receptor (CAR), pregnane X-receptor (PXR), and hepatic xenobiotic detoxifying CYPs in fatty liver disease or obesity. Therefore, we examined whether Cyp3a-null mice show increased obesity and fatty liver disease following 8-weeks of exposure to a 60% high-fat diet (HFD). Surprisingly, HFD-fed Cyp3a-null females fed a HFD gained 50% less weight than wild-type (WT; B6) females fed a HFD. In contrast, Cyp3a-null males gained more weight than WT males, primarily during the first few weeks of HFD-treatment. Cyp3a-null females also recovered faster than WT females from a glucose tolerance test; males showed no difference in glucose tolerance between the groups. Serum concentrations of the anti-obesity hormone, adiponectin are 60% higher and β-hydroxybutyrate levels are nearly 50% lower in Cyp3a-null females than WT females, in agreement with reduced weight gain, faster glucose response, and reduced ketogenesis. In contrast, Cyp3a-null males have higher liver triglyceride concentrations and lipidomic analysis indicates an increase in phosphatidylinositol, phosphatidylserine and sphingomyelin. None of these changes were observed in females. Last, Pxr, Cyp2b, and IL-6 expression increased in Cyp3a-null females following HFD-treatment. Cyp2b and Fatp1 increased, while Pxr, Cpt1a, Srebp1 and Fasn decreased in Cyp3a-null males following a HFD, indicating compensatory biochemical responses in male (and to a lesser extent) female mice fed a HFD. In conclusion, lack of Cyp3a has a positive effect on acclimation to a HFD in females as it improves weight gain, glucose response and ketosis.

Three common false-negative scenarios have been encountered with amendment addition in whole-sediment toxicity identification evaluations (TIEs): dilution of toxicity by amendment addition (i.e., not toxic enough), not enough amendment present to reduce toxicity (i.e., too toxic), and the amendment itself elicits a toxic response (i.e., secondary amendment effect). One such amendment in which all 3 types of false-negatives have been observed is with the nonpolar organic amendment (activated carbon or powdered coconut charcoal). The objective of the present study was to reduce the likelihood of encountering false-negatives with this amendment and to increase the value of the whole-sediment TIE bioassay. To do this, the present study evaluated the effects of various activated carbon additions to survival, growth, emergence, and mean development rate of Chironomus tepperi. Using this information, an alternative method for this amendment was developed which utilized a combination of multiple amendment addition ratios based on wet weight (1%, lower likelihood of the secondary amendment effect; 5%, higher reduction of contaminant) and nonconventional endpoints (emergence, mean development rate). This alternative method was then validated in the laboratory (using spiked sediments) and with contaminated field sediments. Using these multiple activated carbon ratios in combination with additional endpoints (namely, emergence) reduced the likelihood of all 3 types of false-negatives and provided a more sensitive evaluation of risk. Environ Toxicol Chem 2018;37:1219-1230. © 2017 SETAC.

Identifying key toxicants in sediment is a great challenge, particularly if nontarget toxicants are involved. To identify the contaminants responsible for sediment toxicity to Chironomus dilutus in Guangzhou reach of the Pearl River in South China, passive dosing and in vivo toxicity testing were incorporated into effect-directed analysis (EDA) to account for bioavailability. Fractionation of sediment extracts was performed with gel permeation chromatography and reverse phase liquid chromatography sequentially. Polydimethylsiloxane served as passive dosing matrix for midge bioassays. The fractions showing abnormal enzymatic response were subject to a nontarget analysis, which screened out 15 candidate toxicants. The concentrations of the screened contaminants (log-based organic carbon normalized) in sediments of 10 sites were compared to sediment toxicity (10 and 20 day mortality and 10 day enzymatic response) to C. dilutus using correlation analyses. The results suggested that oxidative stress induced by cypermethrin, dimethomorph, pebulate and thenylchlor may have in part caused the observed toxicity to C. dilutus. The present study shows that EDA procedures coupled with passive dosing and in vivo toxicity testing can be effective in identifying sediment-bound toxicants, which may pose high risk to benthic organisms but are not routinely monitored and/or regulated. The findings of the present study highlight the importance of incorporating environmentally relevant approaches in assessing sediment heavily impacted by a multitude of contaminants, which is often the case in many developing countries.

Most of the public literature and available guidance documents on the conduct of freshwater whole-sediment toxicity identification and evaluations (TIEs) detail the use of test organisms and amending agents that are readily available in North America. These commonly used test organisms and the supported amending agents, however, are not available and largely inappropriate (i.e., not native species) for conducting whole-sediment TIEs outside of North America. The overall objective of the present study was to build foundational methods for performing freshwater whole-sediment TIEs in Australia. We examined the capability of 3 amending agents: ANZ38 Zeolite (for ammonia; Castle Mountain Zeolites), Oxpure 325B-9 Activated Carbon (for nonpolar organics; Oxbow Activated Carbon), and Lewatit MonoPlus TP 207 (for cationic metals; Lanxess Deutschland) on 2 Australian native freshwater species: the midge Chironomus tepperi and the amphipod Austrochiltonia subtenuis. To evaluate the effectiveness of each amendment, bioassays were conducted with spiked sediments of ammonia, permethrin (as part of a commercial formulation), and copper using acute median lethal concentrations (LC50s) for both species and growth median effect concentration (EC50) of midges as the endpoints of interest. Environ Toxicol Chem 2017;36:2476-2484. © 2017 SETAC.

During Drosophila embryogenesis, the germband first extends to curl around the posterior end of the embryo and then retracts back; however, retraction is not simply the reversal of extension. At a tissue level, extension is coincident with ventral furrow formation, and at a cellular level, extension occurs via convergent cell neighbor exchanges in the germband, whereas retraction involves only changes in cell shape. To understand how cell shapes, tissue organization, and cellular forces drive germband retraction, we investigate this process using a whole-embryo, surface-wrapped cellular finite-element model. This model represents two key epithelial tissues—amnioserosa and germband—as adjacent sheets of two-dimensional cellular finite elements that are wrapped around an ellipsoidal three-dimensional approximation of an embryo. The model reproduces the detailed kinematics of in vivo retraction by fitting just one free model parameter, the tension along germband cell interfaces; all other cellular forces are constrained to follow ratios inferred from experimental observations. With no additional parameter adjustments, the model also reproduces quantitative assessments of mechanical stress using laser dissection and failures of retraction when amnioserosa cells are removed via mutations or microsurgery. Surprisingly, retraction in the model is robust to changes in cellular force values but is critically dependent on starting from a configuration with highly elongated amnioserosa cells. Their extreme cellular elongation is established during the prior process of germband extension and is then used to drive retraction. The amnioserosa is the one tissue whose cellular morphogenesis is reversed from germband extension to retraction, and this reversal coordinates the forces needed to retract the germband back to its pre-extension position and shape. In this case, cellular force strengths are less important than the carefully established cell shapes that direct them. Video Abstract Download : Download video (40MB)

For the important task of binocular depth perception from complex natural-image stimuli, the neurophysiological basis for disambiguating multiple matches between the eyes across similar features has remained a long-standing problem. Recurrent interactions among binocular disparity-tuned neurons in the primary visual cortex (V1) could play a role in stereoscopic computations by altering responses to favor the most likely depth interpretation for a given image pair. Psychophysical research has shown that binocular disparity stimuli displayed in 1 region of the visual field can be extrapolated into neighboring regions that contain ambiguous depth information. We tested whether neurons in macaque V1 interact in a similar manner and found that unambiguous binocular disparity stimuli displayed in the surrounding visual fields of disparity-selective V1 neurons indeed modified their responses when either bistable stereoscopic or uniform featureless stimuli were presented within their receptive field centers. The delayed timing of the response behavior compared with the timing of classical surround suppression and multiple control experiments suggests that these modulations are carried out by slower disparity-specific recurrent connections among V1 neurons. These results provide explicit evidence that the spatial interactions that are predicted by cooperative algorithms play an important role in solving the stereo correspondence problem.

The present study examined the effects of bioturbation by the oligochaete Lumbriculus variegatus on bioavailability and toxicity of sediment-associated polycyclic aromatic hydrocarbons (PAHs) to the oligochaete and the epi-benthic amphipod Hyalella azteca. Various densities of L. variegatus in sediment were used to represent different levels of bioturbation. Total sediment concentration declined with increasing worm density, but bioavailability of PAHs estimated using the biomimetic extractions showed no significant difference among treatments with different worm densities. Alternatively, PAH bioaccumulation by L. variegatus decreased and the growth of the worms was reduced at the highest worm density, which was probably due to overcrowding of organisms, food competition and increasing release of PAHs by bioturbation. Additionally, co-exposure with high density of L. variegatus to contaminated sediment significantly increased PAH accumulation and mortality of H. azteca. The increased toxicity was probably because of the transport of sediment-associated contaminants to sediment surface and water column by the bioturbation by L. variegatus. Overall, the present study showed that bioturbation may alter the toxicity of contaminants in sediment to other organisms, thus the presence of benthic invertebrates and their interactions with the sediment should be considered in future sediment risk assessment.

Insecticides and heavy metals are frequently detected in the environment, but few studies have assessed the joint toxicity of organic and inorganic contaminants. Joint toxicity of a pyrethroid insecticide, cypermethrin, and a heavy metal, Pb2+, was evaluated in the present study. An antagonistic toxic response was observed when the benthic invertebrate Chironomus dilutus was simultaneously exposed to the two contaminants in both water and sediment exposures. Pre-exposure bioassays with midges were also conducted, and toxicity of cypermethrin was significantly reduced for midges that were pre-exposed to Pb2+. In addition, the impact of Pb2+ on the bioavailability of cypermethrin to midges was measured using Tenax extraction. No significant difference was noted in the amount of Tenax-extractable cypermethrin when different amounts of Pb2+ were added to sediment. Results suggested that altered organism sensitivity may contribute to the observed antagonistic interaction between cypermethrin and Pb2+, whereas the influence of changes in toxicokinetic processes such as uptake, biotransformation, and elimination on the joint toxicity should be further studied. Finally, the decreased toxicity of cypermethrin when simultaneously applied with Pb2+ may be one reason for the overestimation of sediment toxicity by cypermethrin alone in field-collected sediment. Environ. Toxicol. Chem. 2011;30:2838–2845. © 2011 SETAC

A male advantage over females for spatial tasks has been well documented in both humans and rodents, but it remains unclear how the activational effects of testosterone influence spatial ability in males. In a series of experiments, we tested how injections of testosterone influenced the spatial working and reference memory of castrated male rats. In the eight-arm radial maze, testosterone injections (0.500 mg/rat) reduced the number of working memory errors during the early blocks of testing but had no effect on the number of reference memory errors relative to the castrated control group. In a reference memory version of the Morris water maze, injections of a wide range of testosterone doses (0.0625-1.000 mg/rat) reduced path lengths to the hidden platform, indicative of improved spatial learning. This improved learning was independent of testosterone dose, with all treatment groups showing better performance than the castrated control males. Furthermore, this effect was only observed when rats were given testosterone injections starting seven days prior to water maze testing and not when injections were given only on the testing days. We also observed that certain doses of testosterone (0.250 and 1.000 mg/rat) increased perseverative behavior in a reversal-learning task. Finally, testosterone did not have a clear effect on spatial working memory in the Morris water maze, although intermediate doses seemed to optimize performance. Overall, the results indicate that testosterone can have positive activational effects on spatial learning and memory, but the duration of testosterone replacement and the nature of the spatial task modify these effects.

Few studies have been conducted examining the distribution of different-sized particles in sediment and its potential impact on bioavailability of sediment-associated contaminants. In the current study, three sediments composed of different particle sizes, i.e., fine (0-180 μm), combined (0-500 μm), and coarse (180-500 μm), were used to evaluate the bioaccumulation potential and toxicokinetic rates of four hydrophobic organic contaminants (HOCs) including two polychlorinated biphenyls (PCB-101 and PCB-118), a metabolite of an organochlorine insecticide (p,p'-DDE), and a polybrominated diphenyl ether (BDE-47) to the benthic oligochaete Lumbriculus variegatus. Two chemical approaches, Tenax extraction and matrix-solid phase microextraction (SPME), were also used to measure bioavailability of the sediment-associated HOCs. The uptake and elimination rates of HOCs by L. variegatus from coarse sediment were greater than those from fine sediment, although the biota-sediment accumulation factors (BSAFs) were not significantly different among sediments with different particle sizes. The freely dissolved HOC concentrations measured by matrix-SPME were greater in coarse sediment, however, no difference was found in uptake and desorption rates for the matrix-SPME and Tenax extraction measurements. Although BSAFs in L. variegatus were the same among sediments, kinetic rates of HOCs for organisms and freely dissolved HOC concentrations were lower in fine sediment, suggesting that sediment ingestion may also play a role in organism uptake, especially for HOCs in fine sediment.

Porewater and whole sediment toxicity identification evaluations (TIEs) were performed on contaminated Illinois River sediment and compared using two standardized toxicity-testing organisms (Ceriodaphnia dubia and Hyalella azteca). Results suggested that the choice of testing matrix (porewater versus whole sediment) significantly influenced characterization of toxicity. The porewater TIE suggested that ammonia was the major source of toxicity, while the whole sediment TIE indicated that non-polar organics, specifically polycyclic aromatic hydrocarbons, were the primary contributor to toxicity, with ammonia being a secondary contributor to toxicity. While the choice of test organism may have played a smaller role in the discordance between the TIEs, the data suggest that this factor alone could play a prevalent role in characterizing toxicity in other TIE assessments. Because porewater and whole sediment TIEs examine sediment toxicity differently, using both TIE approaches as part of a risk assessment may provide a more accurate risk estimate of sediment toxicity.

Whole-sediment toxicity identification evaluation (TIE) techniques were employed on the Illinois River Complex (IRC), USA to identify the sources of sediment toxicity that may have contributed to the decline in benthic invertebrate populations. The TIE focused on three classes of contaminants: ammonia, metals, and organics. Sediment toxicity was assessed using the amphipod Hyalella azteca, and the focus of the TIE was on assessing spatial and temporal patterns of contamination. Past studies suggested that ammonia was the major source of contamination in IRC sediments. However, the present study suggested that polycyclic aromatic hydrocarbons (PAHs) were the primary contributor to sediment toxicity. Phase I testing showed 46% of the site trials (12 of 26) exhibited increased H. azteca survival (p

Objective—To determine the effect of time interval from birth to first colostrum feeding on colostrum intake and serum IgG concentration and the effect of varying colostral volume intake and colostral IgG concentration on the probability of failure of passive transfer (FPT) in bottle-fed calves. Design—Randomized controlled study. Animals—104 calves. Procedures—Equal numbers of calves were randomly assigned to groups and fed 3 L of their dam's colostrum at 1, 2, 3, or 4 hours after birth by use of a nipple bottle. Calves were allowed to feed for 15 minutes, and intake was recorded. A second 3-L bottle feeding of colostrum was offered at 12 hours of age. Results—17.2% of calves ingested 3 L of colostrum at the first feeding and 3 L at 12 hours of age. Calf age, up to 4 hours, had no significant effect on the calf's ability to ingest colostrum or on 48-hour serum IgG concentration. Colostral intake at 1, 2, 3, or 4 hours had no effect on intake at the second feeding. Probability of FPT in calves ingesting 3 L at both feedings was < 0.05. Conclusions and Clinical Relevance—Allowing calves fed by nipple bottle to ingest as much colostrum as they can within 4 hours after birth and at 12 hours of age substantially reduced the probability of FPT. Bottle-fed calves that do not ingest 3 L of colostrum within the first 4 hours after birth should be targeted for oroesophageal intubation.

The whole body tension (WBT) method was used to evaluate the hypothesis that long term treatment with NF-kappaB inhibitors improves the total forward pulling tension exerted by the limb musculature of the mdx mouse. Mdx mice exhibited significantly reduced WBT values and more profound weakening during the course of generating multiple forward pulling movements than age-matched nondystrophic mice. Long term treatment with the NF-kappaB inhibitor pyrrolidine dithiocarbamate (PDTC) did not significantly reduce nuclear p65 activation in the costal diaphragm, but increased WBT by 12% in mature (12 month) mice. Daily treatment (30 days) of 1 month old mdx mice with the inhibitor ursodeoxycholic acid (UDCA) reduced costal diaphragm nuclear p65 activation by 40% and increased WBT by 21%. These results indicate that treatment with NF-kappaB inhibitors improves WBT in the mdx mouse and further establishes the utility of the WBT procedure in assessing therapeutic efficacy.

Objective—To determine associations between age, sex, breed, and month and year of admission and the diagnosis of lead toxicosis in cattle. Design—Retrospective case-control study. Sample Population—Records of all cattle evaluated at North American veterinary teaching hospitals during the years 1963 to 2002, which were available through the Veterinary Medical Database. Procedures—Logistic regression was used to evaluate the associations between postulated risk factors and the occurrence of lead toxicosis in cattle and predict the occurrence of the diagnosis of lead toxicosis in cattle. Results—413 cases of lead intoxication and 202,363 control cattle were identified and met the inclusion criteria. Cattle < 4 years of age were at increased risk for the diagnosis of lead intoxication relative to cattle ≥ 4 years of age. Cattle ≥ 2 months and < 6 months of age had the greatest risk for lead intoxication (odds ratio, 12.3). Angus cattle were at greater risk for toxicosis (odds ratio, 1.95), compared with other breeds. The risk of lead toxicosis was greater before 1985 (odds ratio, 1.94) than the risk thereafter. The risk of lead toxicosis diagnosis was greatest in the months of May, June, July, and August. Conclusions and Clinical Relevance—Lead toxicosis in cattle was associated with age < 4 years and the Angus breed. A seasonal pattern existed with peak occurrence in the late spring and summer. The occurrence of lead toxicosis has declined over time.

Objective—To evaluate the effect of lactoferrin on lipopolysaccharide (LPS)-induced proliferation of bovine peripheral blood mononuclear cells (PBMCs), gene expression of inflammatory mediators, and production of prostanoids in vitro. Sample Population—PBMCs isolated from 15 Holstein bull calves. Procedures—Mixed populations of PBMCs were isolated by differential centrifugation. Proliferation assays were conducted in 96-well plates designed to allow addition of lactoferrin (200 ng/mL) with and without LPS (1 μg/mL) in a checkerboard design. Incorporation of 3H-thymidine was used to determine proliferation of PBMCs. Prostaglandin E2 production was determined in culture-conditioned medium by use of enzyme immunoassay. Effects of lactoferrin on LPS-induced gene expression of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 were monitored by use of PCR assays. Results—Lactoferrin supplementation significantly reduced LPS-induced incorporation of 3H-thymidine and production of prostaglandin E2 by PBMCs. Lactoferrin reduced LPS-induced expression of COX-2 and MMP-9 mRNA. Conclusions and Clinical Relevance—Lactoferrin reduced LPS-induced cellular proliferation, inflammatory mediator gene expression, and prostaglandin E2 production by bovine PBMCs in vitro. These effects may be beneficial in reducing the impact of endotoxemia in neonates.

Objective—To compare distributions of survivin among tissues from urinary bladders of dogs with cystitis, transitional cell carcinoma (TCC), or histologically normal urinary bladders. Sample Population—24 archived and 7 fresh-frozen specimens of urinary bladders from dogs with cystitis. Procedures—Immunohistochemical analysis of archived tissue specimens was performed to identify survivin protein in the nucleus and cytoplasm of cells by use of polyclonal rabbit anti-survivin antibody. Tissues that contained ≥ 5% immunoreactive cells were considered positive for survivin protein. Reverse-transcription PCR analysis was performed on fresh-frozen tissues to identify survivin mRNA. Data on tissues from dogs with TCC or histologically normal urinary bladders that were obtained during another study were used for statistical comparisons. Results—Twelve of 24 (50%) cystitic tissues were positive for nuclear survivin, compared with 28 of 41 (68%) TCC tissues and 0 of 46 (0%) normal tissues. Two of 24 (8%) cystitic tissues were positive for cytoplasmic survivin, compared with 7 of 41 (17%) TCC tissues and 17 of 46 (37%) normal tissues. Proportions of specimens that contained nuclear or cytoplasmic survivin were significantly different between cystitic and normal tissues but not between cystitic and TCC tissues. Four of 7 cystitic tissues were positive for survivin mRNA, which was comparable with results for TCC and normal tissues. Conclusions and Clinical Relevance—Nuclear survivin was detected in TCC and cystitic tissues but not in normal urinary bladder tissues. Additional studies are needed to determine whether nuclear survivin contributes to the development or progression of TCC.

The joint toxicity of chlorpyrifos and atrazine was compared to that of chlorpyrifos alone to discern any greater than additive response using both acute toxicity testing and whole-body residue analysis. In addition, acetylcholinesterase (AChE) inhibition and biotransformation were investigated to evaluate the toxic mode of action of chlorpyrifos in the presence of atrazine. The joint toxicity of atrazine and chlorpyrifos exhibited no significant difference in Lepomis macrochirus compared to chlorpyrifos alone; while studies performed with Pimephales promelas and Chironomus tentans, did show significant differences. AChE activity and biotransformation showed no significant differences between the joint toxicity of atrazine and chlorpyrifos and that of chlorpyrifos alone. From the data collected, the combination of atrazine and chlorpyrifos pose little additional risk than that of chlorpyrifos alone to the tested fish species.

Objective—To characterize the biological behavior and prognostic factors associated with hemangiosarcoma in cats. Design—Retrospective case series. Animals—53 cats with hemangiosarcoma. Procedures—Data were retrieved from a state veterinary diagnostic laboratory, 3 veterinary colleges, and a private practice. Results—Cutaneous and subcutaneous tumor locations were more common than visceral (abdominal and thoracic) and oral locations. Surgical excision was the primary treatment in 47 cats. Tumor-free surgical margins were more likely in cutaneous than subcutaneous lesions and were associated with longer survival times. Local recurrence was observed in 6 of 12 cats with subcutaneous lesions for which follow-up was available. Metastatic disease was detected in 5 of 13 cats with adequate staging at initial diagnosis. A sixth cat had pulmonary metastases at the time of euthanasia. In 4 of 10 cats with visceral hemangiosarcoma, the diagnosis was made at necropsy or they were euthanized at the time of diagnosis. Adjuvant therapy was uncommonly used. Eighteen of the 21 known deaths or euthanasias were tumor-related. Higher mitotic counts (> 3 in 10 hpfs) were associated with shorter survival times. Conclusions and Clinical Relevance—Subcutaneous hemangiosarcoma was more biologically aggressive than the cutaneous form and was more likely to recur locally and result in euthanasia or death of the cat. Metastatic potential of the cutaneous and subcutaneous forms may be greater than previously reported. Visceral hemangiosarcoma is associated with a grave prognosis.

Objective—To characterize features and response to treatment of keratoconjunctivitis sicca (KCS) associated with oral administration of etodolac in dogs. Design—Retrospective case series. Sample Population—65 cases obtained from a survey of veterinary ophthalmologists (group A) and 146 cases reported to Fort Dodge Animal Health (group B). Procedures—Data analyzed included breed, sex, age, weight, dose and duration of etodolac administration, results of Schirmer tear test at the time of diagnosis and last follow-up, treatments, and response to treatments. Groups A and B were analyzed separately by use of forward stepwise logistic regression models developed to predict probability of complete remission or clinical improvement as a function of several variables. Results—Most dogs developed severe KCS (84 eyes of 50 dogs [group A]; 111 eyes of 62 dogs [group B]). Resolution of KCS occurred in 7 of 65 (A) and 23 of 146 (B) dogs. No response to treatment was observed in 26 of 65 (A) and 27 of 146 (B) dogs. Fifty-one (A) and 52 (B) dogs had records that were sufficiently complete to use in models. In group B, dogs with etodolac treatment intervals < 6 months prior to the onset of KCS were 4.2 times as likely to have remission as were dogs with treatment intervals ≥ 6 months. Conclusions and Clinical Relevance—Shorter duration of etodolac administration (< 6 months) was associated with improved outcome in 1 population of dogs. Monitoring of tear production should be considered prior to and during administration of etodolac in dogs.

Reported clinical signs of coccidiosis in South American camelids include anorexia of a few days duration, sudden death, and diarrhea. Antemortem diagnosis of clinical coccidiosis is usually based on clinical signs and supported by detection of coccidial oocysts in feces. This report describes 2 atypical cases of coccidiosis in South American camelids that had no coccidial oocysts detected on antemortem fecal flotation, prolonged weight loss, and normal fecal consistency.

Heat shocks applied at the onset of gastrulation in early Drosophila embryos frequently lead to phenocopies of U-shaped mutants-having characteristic failures in the late morphogenetic processes of germband retraction and dorsal closure. The pathway from nonspecific heat stress to phenocopied abnormalities is unknown.Drosophila embryos subjected to 30-min, 38 °C heat shocks at gastrulation appear to recover and restart morphogenesis. Post-heat-shock development appears normal, albeit slower, until a large fraction of embryos develop amnioserosa holes (diameters 100 µm). These holes are positively correlated with terminal U-shaped phenocopies. They initiate between amnioserosa cells and open over tens of minutes by evading normal wound healing responses. They are not caused by tissue-wide increases in mechanical stress or decreases in cell-cell adhesion, but instead appear to initiate from isolated apoptosis of amnioserosa cells.The pathway from heat shock to U-shaped phenocopies involves the opening of one or more large holes in the amnioserosa that compromise its structural integrity and lead to failures in morphogenetic processes that rely on amnioserosa-generated tensile forces. The proposed mechanism by which heat shock leads to hole initiation and expansion is heterochonicity, i.e., disruption of morphogenetic coordination between embryonic and extra-embryonic cell types.

Comprehensive neonatal disease control programs use estrous synchronization and artificial insemination to breed a high percentage of replacement heifers to bulls with high-accuracy expected progeny differences (EPD) for acceptable birth weight. Reducing the incidence of dystocia improves passive transfer of antibodies to calves and reduces likelihood of a prolonged postpartum interval to cycling resumption, allowing breeding of first-calf heifers early in the subsequent season. Use of mature cow-herd estrous synchronization systems, nutritional management for optimum body condition, heifer development plans assuring that heifers are cycling before breeding, and breeding soundness examination (BSE) of bulls ensures that a high percentage of cows conceive early, thereby reducing calves exposed to significantly older calves and amplified pathogen loads during the neonatal period.

Urotensin II (UII) is a cyclic neuropeptide with strong vasoconstrictive activity in the peripheral vasculature. UII receptor mRNA is also expressed in the CNS, in particular in cholinergic neurons located in the mesopontine tegmental area, including the pedunculopontine tegmental (PPT) and lateral dorsal tegmental nuclei. This distribution suggests that the UII system is involved in functions regulated by acetylcholine, such as the sleep-wake cycle. Here, we tested the hypothesis that UII influences cholinergic PPT neuron activity and alters rapid eye movement (REM) sleep patterns in rats. Local administration of UII into the PPT nucleus increases REM sleep without inducing changes in the cortical blood flow. Intracerebroventricular injection of UII enhances both REM sleep and wakefulness and reduces slow-wave sleep 2. Intracerebroventricular, but not local, administration of UII increases cortical blood flow. Moreover, whole-cell recordings from rat-brain slices show that UII selectively excites cholinergic PPT neurons via an inward current and membrane depolarization that were accompanied by membrane conductance decreases. This effect does not depend on action potential generation or fast synaptic transmission because it persisted in the presence of TTX and antagonists of ionotropic glutamate, GABA, and glycine receptors. Collectively, these results suggest that UII plays a role in the regulation of REM sleep independently of its cerebrovascular actions by directly activating cholinergic brainstem neurons.

Cytauxzoon felis produces a disease in domestic cats in the Midwest (U.S.A.), which often leads to a fatal outcome. Although the clinical disease process is well described, there are still many unanswered questions about this organism. For example, it is unknown whether species of ticks other than Dermacentor variabilis can serve as vectors for transmission. With recent reports of surviving cats from limited geographic areas, another relevant question is the potential for genetically less virulent organism strains. This study evaluated 352 individual or pooled tick samples (1,362 total ticks) for the presence of C. felis small subunit ribosomal RNA and internal transcribed spacer 1 (ITS-1) region genes using a polymerase chain reaction (PCR). These ticks were collected from dogs and cats in several Missouri counties, including 10 from cats diagnosed with cytauxzoonosis. Only 3 positive C. felis samples were identified in Amblyomma americanum nymphs, and there was very limited genetic variation noted in both genes. The small number of positive samples did not allow the study to determine which PCR analysis was more sensitive. This is the first known report of ITS-1 gene identification and sequencing for C. felis. It is also the first published investigation of genetic variation in C. felis.

Humans often create and appreciate visual symmetry in their environment, and the underlying brain mechanisms have been a topic of increasing interest. Here, symmetric versus random dot stimuli produced robust functional MRI (fMRI) activity in higher-order regions of human visual cortex (especially areas V3A, V4, V7, and LO) but little activity elsewhere in brain. This fMRI response was found both with and without attention controls. Moreover, it was highly correlated with the psychophysical perception of symmetry. Similar symmetry responses were found by using line-based and dot stimuli and were found at a wide range of stimulus sizes and geometric configurations. Weaker symmetry responses were found in analogous regions of macaque visual cortex by using fMRI techniques with higher sensitivity. This evidence suggests that visual symmetry is specifically enhanced in the human brain, but that the underlying neural mechanisms may nevertheless be resolvable in nonhuman primates.

Objective—To characterize gelatinases in bronchoalveolar lavage fluid (BALF) and gelatinases produced by alveolar macrophages of healthy calves. Sample Population—Samples of BALF and alveolar macrophages obtained from 20 healthy 2-month-old calves. Procedure—BALF was examined by use of gelatin zymography and immunoblotting to detect gelatinases and tissue inhibitor of metalloproteinase (TIMP)-1 and -2. Cultured alveolar macrophages were stimulated with lipopolysaccharide (LPS), and conditioned medium was subjected to zymography. Alveolar macrophage RNA was used for reverse transcriptasepolymerase chain reaction assay of matrix metalloproteinases (MMPs), cyclooxygenase-2, and inducible nitric oxide synthase. Results—Gelatinolytic activity in BALF was evident at 92 kd (14/20 calves; latent MMP-9) and 72 kd (18/20; latent MMP-2). Gelatinolytic activity was evident at 82 kd (10/20 calves; active MMP-9) and 62 kd (17/20; active MMP-2). Gelatinases were inhibited by metal chelators but not serine protease inhibitors. Immunoblotting of BALF protein and conditioned medium confirmed the MMP-2 and -9 proteins. Endogenous inhibitors (ie, TIMPs) were detected in BALF from all calves (TIMP-1) or BALF from only 4 calves (TIMP-2). Cultured alveolar macrophages expressed detectable amounts of MMP-9 mRNA but not MMP-2 mRNA. Conclusions and Clinical Relevance—Healthy calves have detectable amounts of the gelatinases MMP-2 and -9 in BALF. Endogenous inhibitors of MMPs were detected in BALF (ie, TIMP-1, all calves; TIMP-2, 4 calves). Lipopolysaccharide-stimulated alveolar macrophages express MMP-9 but not MMP-2 mRNA. The role of proteases in the pathogenesis of lung injury associated with pneumonia has yet to be determined. (Am J Vet Res 2004;65:163–172)

Objective—To evaluate 3 refractometers for detection of failure of passive transfer (FPT) of immunity in calves, and assess the effect of refractometric test endpoints on sensitivity, specificity, and proportion of calves classified correctly with regard to passive transfer status. Design—Prospective study. Animals—90 calves. Procedure—Blood samples were obtained from calves that were < 10 days old. Serum IgG concentration was determined by use of a radial immunodiffusion assay. Accuracy of 3 refractometers in the prediction of serum IgG concentration was determined by use of standard epidemiologic methods and a linear regression model. Results—At a serum protein concentration test endpoint of 5.2 g/dL, sensitivity of each refractometer was 0.89 or 0.93, and specificity ranged from 0.80 to 0.91. For all refractometers, serum protein concentration test endpoints of 5.0 or 5.2 g/dL resulted in sensitivity > 0.80, specificity > 0.80, and proportion of calves classified correctly > 0.85. Serum protein concentrations equivalent to 1,000 mg of IgG/dL of serum were 4.9, 4.8, and 5.1 g/dL for the 3 refractometers. Conclusions and Clinical Relevance—The refractometers, including a nontemperature-compensating instrument, performed similarly in detection of FPT. Serum protein concentration test endpoints of 5.0 and 5.2 g/dL yielded accurate results in the assessment of adequacy of passive transfer; lower or higher test endpoints misclassified larger numbers of calves. (J Am Vet Med Assoc 2002;221:1605–1608)