Interrelationships of dominance and territorial behaviour in the pupfish,Cyprinodon variegatus

Under laboratory conditions, Cyprinodon variegatus establishes either a dominance or territorial mating system. In the dominance system, one male usually has complete access to receptive females, while in the territorial system, several males have equal access. Four factors were observed to enhance a subdominant's ability to establish a territory and improve his reproductive success; increasing the number of conspecific intruders, increasing the available area, the presence of partial barrier, and the presence of a receptive female.     

Evaluation of pre‐ and post‐zygotic mating barriers,hybrid fitness and phylogenetic relationship between Cyprinodon variegatus variegatus and Cyprinodon variegatus hubbsi (Cyprinodontiformes,Teleostei)

The euryhaline fish Cyprinodon variegatus variegatus (Cvv) is capable of tolerating ambient salinities ranging from 0.3 to 167 g l^?1, but incapable of long‐term survival in freshwater (< 2 mM Na⁺). However, a population of this species, now designated as a subspecies (Cyprinodon variegatus hubbsi; Cvh), has been isolated in several freshwater (0.4–1 mM Na⁺) lakes in central Florida for the past ~150 ky. We previously demonstrated that Cvh has a significantly higher affinity for Na⁺ uptake suggesting that it has adapted to its dilute freshwater environment. We here evaluate whether Cvh should be considered a separate species by characterizing pre‐ and post‐zygotic isolation, Na⁺ transport characteristics of the two populations and their hybrids, and developing a molecular phylogeny of Cvv and Cvh populations in Florida using mtDNA sequence data. We found evidence of partial prezygotic isolation with Cvv females mating almost exclusively (89%) with con‐specific males in choice mating experiments. Partial post‐zygotic isolation was also observed with significant (59–89%) reductions in hatching success of hybrid embryos compared with con‐specific embryos. Na⁺ uptake kinetics in hybrids (both Cvv x Cvh and Cvh x Cvv) bred and raised under common garden conditions were intermediate to Cvh (high affinity) and Cvv (low affinity) indicating that observed differences are genetically based. Similar observations were made with respect to short‐term (96 h) survival of juveniles acutely transferred from 7 mM Na⁺ to a range of more dilute (0.1–2 mM Na⁺) freshwater. Finally, although phylogenetic analysis of Cvv and Cvh populations using mtDNA sequence for ND2 were unable to fully resolve a polytomy between Cvh and Cvv populations from northeastern Florida, these data do not falsify the hypothesis that Cvh is of monophyletic origin. Overall, the available data suggest that Cvh should be considered a separate species or at a minimum an evolutionarily significant unit.     

Postzygotic incompatibilities between the pupfishes, Cyprinodon elegans and Cyprinodon variegatus: hybrid male sterility and sex ratio bias

I examined the intrinsic postzygotic incompatibilities between two pupfishes, Cyprinodon elegans and Cyprinodon variegatus. Laboratory hybridization experiments revealed evidence of strong postzygotic isolation. Male hybrids have very low fertility, and the survival of backcrosses into C. elegans was substantially reduced. In addition, several crosses produced female-biased sex ratios. Crosses involving C. elegans females and C. variegatus males produced only females, and in backcrosses involving hybrid females and C. elegans males, males made up approximately 25% of the offspring. All other crosses produced approximately 50% males. These sex ratios could be explained by genetic incompatibilities that occur, at least in part, on sex chromosomes. Thus, these results provide strong albeit indirect evidence that pupfish have XY chromosomal sex determination. The results of this study provide insight on the evolution of reproductive isolating mechanisms, particularly the role of Haldane's rule and the 'faster-male' theory in taxa lacking well-differentiated sex chromosomes.     

Osmotic regulation in the Sheepshead Minnow Cyprinodon variegatus Lacépède

Plasma osmotic regulation was evaluated in Cyprinodon variegatus acclimated to salinities from fresh water to 120%. These fish showed roughly stable regulation in the range from 0.3% to 70%. Fish acclimated to fresh water showed an increased plasma concentration over those in 0.3%, thought to result from a higher Ca⁺⁺ level in fresh water. Plasma concentrations rose abruptly at salinities above 70%. Osmotic regulatory characteristics of C. variegatus were compared with data for other extremely euryhaline species with expectation of a graded series of responses, species showing greatest fresh water tolerance demonstrating widest ranges of regulation and with lowest plasma concentrations. Results were not consistent with expectations. The range of ambient salinities over which good regulation of plasma concentration was shown was found to be directly correlated with the magnitude of the salinity tolerance range of all of the species compared. Forms with greater fresh water affinity generally, but not uniformly, showed the lowest plasma concentrations in the region of hyperosmotic regulation.     

Effects of fluoxetine exposure on serotonin-related activity in the sheepshead minnow (Cyprinodon variegatus) using LC/MS/MS detection and quantitation

Fluoxetine (FLX) is a selective serotonin reuptake inhibitor and is among the top 100 drugs prescribed yearly in the United States and the United Kingdom. Tissue and water extraction methods were developed to detect and quantify FLX, norfluoxetine and the associated biological compounds serotonin (5-HT), 5-hydroxyindole-3-acetic acid (5-HIAA), tryptophan (TRP) and melatonin (MEL) using LC/MS/MS. Acute mortality and sublethal physiological effects of FLX were assessed using standard static renewal toxicity tests in which juvenile sheepshead minnows (Cyprinodon variegatus) were exposed to FLX. Fluoxetine did not cause significant mortality at levels near currently reported environmental concentrations. Significant changes in neurotransmitter levels were observed within the serotonergic system in juvenile sheepshead minnows exposed at concentrations approximately one order of magnitude above those currently reported in the environment. Transformation activity ratios of a product to a precursor compound (5-HT/TRP, 5-HIAA/5-HT and MEL/5-HT) also exhibited significant changes with FLX treatment. Fluoxetine exposure did not only affect 5-HT but had additional effects both upstream and downstream of 5-HT within its synthesis and metabolic pathways. These sublethal changes within the serotonergic pathway may result in behavioral changes which could, in turn, have implications for the ecological response of populations to additional environmental stressors.     

The influence of ambient salinity on routine metabolism in the teleost Cyprinodon variegatus Lacepède

Routine metabolism of the euryhaline cyprinodontid Cyprinodon variegatus Lacepede was measured at a series of ambient salinities ranging from fresh water through 100‰. Fish used had been sequentially acclimated to the test salinities (ambient temperature of 20±1° C and a 12: 12 L: D schedule).
Routine metabolic rates were highest at ambient salinities from 15 to 50‰. Metabolism was somewhat lower at ambient salinities less than 15‰, and showed a sequential decline at ambient salinities greater than 50‰. It is suggested that routine metabolism is depressed at elevated salinities by reduced O, transfer, a consequence of maintenance of hydromineral balance in hypersaline waters.     

Experimental influence of pH on the early life-stages of sea urchins II: increasing parental exposure times gives rise to different responses

Many studies into the responses of early life-stages to ocean acidification utilise offspring obtained from parents reared under present-day conditions. Their offspring are directly introduced to altered-pH conditions. This study determined whether this approach is suitable by pre-exposing parent sea urchins (Psammechinus miliaris) to altered seawater pH (~1000?μatm) for several durations, spawning them and rearing their offspring to settlement. Parents acclimated when exposed to low seawater pH for extended periods (>42?d). Longer adult pre-exposures reduced larval survival and less competent offspring were removed from populations earlier than in controls. Control offspring were larger during earlier development stages (2–7?d), but smaller during later development stages (14?+?d) than offspring reared under low pH conditions. Juvenile settlement levels were similar across all treatments. After 17?d, offspring sourced from parents pre-exposed to low pH for 42 and 70?d were larger than those pre-exposed for 28?d and ambient sourced offspring directly transferred to low pH. These different responses show that the use of ambient derived offspring utilised in many studies is likely not an ideal approach when assessing larval development responses via morphometric measurements and survivorship prior to settlement. This study also suggests that calcifying organisms have capacities to acclimate and possibly adapt towards conditions beyond natural rates of ocean acidification.     

Harmful dinoflagellate Cochlodinium polykrikoides impairs the feeding behavior of larval sheepshead minnows (Cyprinodon variegatus)

Research evaluating the toxicity of the harmful dinoflagellate Cochlodinium (a.k.a. Margalefidinium) polykrikoides has been dominated by acute bioassays while the sublethal effects remain less well understood. This study examined the sublethal effects of C. polykrikoides exposure on the feeding behavior of larval estuarine fish. Sheepshead minnow (Cyprinodon variegatus) larvae were used in feeding experiments which assessed the total consumption of zooplankton prey (i.e., Artemia nauplii) over defined time periods. Larvae exposed to intermediate concentrations (i.e., 10² cells ml⁻¹) of clonal cultures of C. polykrikoides saw statistically significant reductions (range = 10%–81%) in the Artemia consumed compared to controls (i.e., filtered seawater, culture media or nontoxin producing dinoflagellate). These reductions were found independent of whether the larvae were fed or starved prior to experimentation. As these concentrations are similar to those typically found during mild blooms or at the periphery of dense blooms, these findings have significant implications for the feeding behavior of ichthyoplankton.     

Generation,analysis and functional annotation of expressed sequence tags from the sheepshead minnow (Cyprinodon variegatus)

Background

Genomic islands (GIs) are clusters of alien genes in some bacterial genomes, but not be seen in the genomes of other strains within the same genus. The detection of GIs is extremely important to the medical and environmental communities. Despite the discovery of the GI associated features, accurate detection of GIs is still far from satisfactory.

Results

In this paper, we combined multiple GI-associated features, and applied and compared various machine learning approaches to evaluate the classification accuracy of GIs datasets on three genera: Salmonella, Staphylococcus, Streptococcus, and their mixed dataset of all three genera. The experimental results have shown that, in general, the decision tree approach outperformed better than other machine learning methods according to five performance evaluation metrics. Using J48 decision trees as base classifiers, we further applied four ensemble algorithms, including adaBoost, bagging, multiboost and random forest, on the same datasets. We found that, overall, these ensemble classifiers could improve classification accuracy.

Conclusions

We conclude that decision trees based ensemble algorithms could accurately classify GIs and non-GIs, and recommend the use of these methods for the future GI data analysis. The software package for detecting GIs can be accessed at http://www.esu.edu/cpsc/che_lab/software/GIDetector/.

     

Phenology of recruitment and infection patterns of Ascocotyle pachycystis, a digenean parasite in the sheepshead minnow, Cyprinodon variegatus

In northwest Florida, Cyprinodon variegatus are parasitized by Ascocotyle pachycystis, a digenean parasite that forms metacercarial cysts in the lumen of the bulbus arteriosus. Field experiments revealed that fish accumulated parasites at an uneven rate within the highly seasonal trematode recruitment period. Older (= larger) fish had higher rates of parasite recruitment and higher parasite prevalences and densities (numbers of metacercariae per individual fish) than did younger (= smaller) fish. Nearly all adults were parasitized (prevalence range 70-100%), and parasite densities ranged from zero to 6800 per fish. Parasite distributions were clumped (= aggregated) in fish of all age classes but were less heterogeneous in early juveniles and adults than they were in late juveniles. Parasites affected the population dynamics of sheepshead minnows by causing reduced winter survivorship, as evidenced by an increase in the average size of fish and a decrease in the average infection intensity over the winter. This revised version was published online in July 2006 with corrections to the Cover Date.     

Mechanisms involved in the neurotoxic and cognitive effects of developmental methamphetamine exposure

Methamphetamine exposure in utero leads to a variety of higher‐order cognitive deficits, such as decreased attention and working, and spatial memory impairments in exposed children (Piper et al., 2011; Roussotte et al., 2011; Kiblawi et al., 2011). As with other teratogens, the timing of methamphetamine exposure greatly determines its effects on both neuroanatomical and behavioral outcomes. Methamphetamine exposure in rodents during the third trimester human equivalent period of brain development results in distinct and long‐lasting route‐based and spatial navigation deficits (Williams et al., 2003; Vorhees et al., 2005, 2008, 2009;). Here, we examine the impact of neonatal methamphetamine‐induced neurotoxicity on behavioral outcomes, neurotransmission, receptor changes, plasticity proteins, and DNA damage. Birth Defects Research (Part C) 108:131–141, 2016. © 2016 Wiley Periodicals, Inc.     

Behavioral and reproductive effects of chronic developmental exposure to brominated vegetable oil in rats

Adult Sprague-Dawley rats were fed diets containing 0, 0.25, 0.5, 1.0, or 2.0% of the food additive brominated vegetable (soybean) oil (BVO) for 2 weeks prior to mating. After conception, the diets were continued throughout gestation and lactation for the females. The same diets were also provided to the dams' offspring throughout their development (up to 90-120 days of age). BVO at 2.0% of the diet completely blocked reproduction. BVO at 1.0% of the diet severely impaired conception, reduced maternal body weight, and produced slightly reduced litter sizes but no evidence of malformations. At this dose postnatal mortality was high, and survivors showed impaired growth and severe behavioral impairments on a battery of standardized tests of functional development. After weaning, adequate data could not be obtained because of the high mortality rate in this group. BVO at 0.5% of the diet produced less reproductive interference and much less offspring mortality or impairment of growth, but produced behavioral impairments almost as severe as seen in the BVO 1.0% group. In addition, this group exhibited severely reduced postweaning activity, delayed vaginal patency development, and reduced day-90 weight. BVO at 0.25% of the diet produced reproductive deficits similar to the BVO 0.5% group, but less severe effects on growth and behavioral development. This group showed no significant increase in offspring mortality. The data demonstrate clear evidence of dose-related physical and behavioral developmental toxicity.     

Toxic effects of in vitro exposure to p-tert-octylphenol on bovine oocyte maturation and developmental competence

Alkylphenolic compounds are a widespread family of xenoestrogens. High concentrations of these substances are present in sewage sludge that is spread on arable land and pasture as fertilizer. Because of their known endocrine system-disrupting activity, alkylphenols represent a potential risk for the reproductive health of farm animals. In this study, the impact of p-tert-octylphenol (OP) on the developmental competence of bovine oocytes was evaluated. Endocrine activity of OP was investigated for its effect on estrogen receptors alpha and beta (ERalpha and ERbeta) and progesterone receptor (PR) mRNA levels. Cumulus-oocyte complexes (COCs) were exposed during in vitro maturation to serial concentrations of OP (1-0.0001 microg/ml) and were compared with vehicle-treated controls and a group of COCs treated with 17 beta-estradiol (E2). A dose-related decrease in the percentage of oocytes that completed maturation after 24 h and in oocyte fertilization competence was observed at doses of OP as low as 0.01 microg/ml. Groups treated with > or =0.001 microg/ml OP showed impaired embryo development. No adverse effects of E2 were observed. In the E2-treated COCs, ERalpha mRNA was decreased but PR mRNA was upregulated compared with controls. Treatment with 0.001 and 0.0001 microg/ml OP induced a decrease in ERalpha mRNA, but ERbeta and PR mRNA were not affected. Treatment with 0.01 microg/ml OP did not produce changes in the expression of any of the mRNAs studied. OP impairs meiotic progression and developmental competence of bovine oocytes without demonstrating clear estrogen-mimic activity.     

Acetylcholinesterase in the sea urchin Lytechinus variegatus: characterization and developmental expression in larvae

Acetylcholinesterase (AChE) in the echinoid Lytechinus variegatus has been characterized. Kinetic parameters V(max), K(m), K(ss), and b were 2594+/-1048 nmol ATCh hydrolyzed/min/mg tissue wet weight, 185+/-11 microM, 308+/-100 mM, and 0.2, respectively for the substrate ATCh and 17.8+/-6.87 nmol BTCh hydrolyzed/min/mg tissue wet weight, 654+/-424 microM, 36+/-31 mM, and 0.6, respectively for BTCh. Pharmacologic analyses were performed with four inhibitors of cholinesterases, physostigmine, BW284c51, ethopropazine, and iso-OMPA, and yielded IC(50) values of 106+/-4 nM, 718+/-118 nM, 2.57+/-0.6 mM, and >0.0300 M, respectively. Both kinetic and pharmacologic results confirmed the existence of AChE in larval L. variegatus. Dimeric and tetrameric globular forms (determined by velocity sedimentation on sucrose gradients) were present in L. variegatus larvae. Activity of AChE increased significantly as larvae progressed in development from embryos to eight-arm larvae. Acetylcholinesterase activity of F1 larvae derived from sea urchins collected from wild populations and of F1 larvae derived from sea urchins cultured in the laboratory and fed two different diets suggest that the nutritional and/or environmental history of the adult sea urchin affect the developmental progression of AChE activity in the F1 offspring.     

Molecular underpinnings of the early brain developmental response to differential feeding in the honey bee Apis mellifera

Brain differential morphogenesis in females is one of the major phenotypic manifestations of caste development in honey bees. Brain diphenism appears at the fourth larval phase as a result of the differential feeding regime developing females are submitted during early phases of larval development. Here, we used a forward genetics approach to test the early brain molecular response to differential feeding leading to the brain diphenism observed at later developmental phases. Using RNA sequencing analysis, we identified 53 differentially expressed genes (DEGs) between the brains of queens and workers at the third larval phase. Since miRNAs have been suggested to play a role in caste differentiation after horizontal and vertical transmission, we tested their potential participation in regulating the DEGs. The miRNA-mRNA interaction network, including the DEGs and the royal- and worker-jelly enriched miRNA populations, revealed a subset of miRNAs potentially involved in regulating the expression of DEGs. The interaction of miR-34, miR-210, and miR-317 with Takeout, Neurotrophin-1, Forked, and Masquerade genes was experimentally confirmed using a luciferase reporter system. Taken together, our results reconstruct the regulatory network that governs the development of the early brain diphenism in honey bees.     

Behavioral effects of developmental methylmercury drinking water exposure in rodents

Early methylmercury (MeHg) exposure can have long-lasting consequences likely arising from impaired developmental processes, the outcome of which has been exposed in several longitudinal studies of affected populations. Given the large number of newborns at an increased risk of learning disabilities associated with in utero MeHg exposure, it is important to study neurobehavioral alterations using ecologically valid and physiologically relevant models. This review highlights the benefits of using the MeHg drinking water exposure paradigm and outlines behavioral outcomes arising from this procedure in rodents. Combination treatments that exacerbate or ameliorate MeHg-induced effects, and possible molecular mechanisms underlying behavioral impairment are also discussed.     

Molecular targets and early response biomarkers for the prediction of developmental toxicity in vitro

There is an urgent need for new in vitro methods to predict the potential developmental toxicity of candidate drugs in the early lead identification and optimisation process. This would lead to a reduction in the total number of animals required in full-scale developmental toxicology studies, and would improve the efficiency of drug development. However, suitable in vitro systems permitting robust high-throughput screening for this purpose, for the most part, remain to be designed. An understanding of the mechanisms involved in developmental toxicity may be essential for the validation of in vitro tests. Early response biomarkers - even a single one - could contribute to reducing assay time and facilitating automation. The use of toxicogenomics approaches to study in vitro and in vivo models in parallel may be a powerful tool in defining such mechanisms of action and the molecular targets of toxicity, and also for use in finding possible biomarkers of early response. Using valproic acid as a model substance, the use of DNA microarrays to identify teratogen-responsive genes in cell models is discussed. It is concluded that gene expression in P19 mouse embryocarcinoma cells represents a potentially suitable assay system, which could be readily used in a tiered testing system for developmental toxicity testing.     

Single exposure to radiation produces early anti-angiogenic effects in mouse aorta

Radiation exposure can increase the risk for many non-malignant physiological complications, including cardiovascular disease. We have previously demonstrated that ionizing radiation can induce endothelial dysfunction, which contributes to increased vascular stiffness. In this study, we demonstrate that gamma radiation exposure reduced endothelial cell viability or proliferative capacity using an in vitro aortic angiogenesis assay. Segments of mouse aorta were embedded in a Matrigel-media matrix 1 day after mice received whole-body gamma irradiation between 0 and 20 Gy. Using three-dimensional phase contrast microscopy, we quantified cellular outgrowth from the aorta. Through fluorescent imaging of embedded aortas from Tie2GFP transgenic mice, we determined that the cellular outgrowth is primarily of endothelial cell origin. Significantly less endothelial cell outgrowth was observed in aortas of mice receiving radiation of 5, 10, and 20 Gy radiation, suggesting radiation-induced endothelial injury. Following 0.5 and 1 Gy doses of whole-body irradiation, reduced outgrowth was still detected. Furthermore, outgrowth was not affected by the location of the aortic segments excised along the descending aorta. In conclusion, a single exposure to gamma radiation significantly reduces endothelial cell outgrowth in a dose-dependent manner. Consequently, radiation exposure may inhibit re-endothelialization or angiogenesis after a vascular injury, which would impede vascular recovery.     

The Pleistocene history of the sheepshead minnow (Cyprinodon variegatus): Non-equilibrium evolutionary dynamics within a diversifying species complex

The sheepshead minnow, Cyprinodon variegatus, is a widespread fish species that typically inhabits coastal tidal marsh and mangrove swamp environments, ranging from Cape Cod, Massaschusetts to northern Mexico and into the Caribbean. This wide range crosses several biogeographic boundaries which are coincident with genetic structuring within numerous species originating in the Pleistocene. In addition, the more northerly reaches of this species range have been further subject to the evolutionary consequences of Pleistocene glaciation due to local extinction and recolonization of formerly glaciated sites. C. variegatus thus provides an excellent vertebrate model system within which to test the extent of genetic differentiation among populations in a dominant coastal ecosystem and examine patterns of historical demography in populations distributed along a latitudinal gradient. Using mitochondrial control region and ND2 sequence data, we discovered monophyletic clades within C. variegatus with divergence times within the Pleistocene, and very low gene flow between most sites. Intraspecific genetic breaks appear to correspond broadly to biogeographic or oceanic boundaries. Pleistocene climate change appears to have had dramatic impacts on the size and distribution of populations within and near the glacial margins, but has also affected populations far from formerly glaciated regions.