Reproductive biology of hardhead catfish Ariopsis felis: evidence for overwintering oocytes.

Hardhead catfish Ariopsis felis are a common marine catfish in the coastal waters of the Gulf of Mexico (GOM). The low economic value of this species has depressed interest and research, and although the species is known for its extremely low fecundity and large oocytes, little else is known about this catfish species. A total of 1230 samples across all months of the year from 2016 to 2018 resulted in 681 females, and analysis of gonado-somatic index (IG ) revealed 1% to be a clear cut-off indicating maturity. Females are considered capable of spawning from April to June when IG averaged 4-8%. Both atresia and post-ovulatory follicles were present in July, suggesting that spawning ends in July in the northern GOM. The 1% IG cut-off was used to designate maturity, and from that an L50 of 253 mm was estimated. Batch fecundity from 41 females estimated a mean batch size of 36 oocytes. Perhaps the most interesting finding was the presence of secondary growth stage oocytes (e.g., cortical alveoli) from July through November, well outside the spawning capable period. Furthermore, 78% of females had some early vitellogenic oocytes present during the non-spawning season, and the distribution of these relatively large (2-5 mm) oocytes did not change over time. The results here are not only important as reproductive biology information for a common and abundant species, but also present interesting and unusual patterns of non-spawning season oocyte development that is not commonly seen in Western Hemisphere subtropical fish species.

Characterization of prostate cancer cell progression in zebrafish xenograft model.

Early diagnosis of prostate cancer (PCa) is critical for the application of efficient treatment to PCa patients. However, the majority of PCas remains indolent from several months to several years before malignancy. Current diagnosis methods have limitations in their reliability and are inefficient in time cost. Thus, an efficient in vivo PCa cell xenograft model is highly desired for diagnostic studies in PCas. In the present study we present a standardized procedure to create a PCa cell xenograft model using zebrafish (Danio rerio) as the host. PC3-CTR cells, a cell line from adenocarcinoma with stable expression of calcitonin receptor (CRT), were subcutaneously injected into zebrafish larvae at 48 h post fertilization. The nursing conditions for the larvae were optimized with stable survival rates of post hatch and post PC3-CTR cell injection. In this system, the progression of PC3-CTR cells in vivo was evaluated by migration and proliferation of the cells. Massive migrations of PC3 cells in vivo were observed at post injection day (PID)3. The injected PC3-CTR cells eventually invaded the whole larval zebrafish at PID5. Quantification of PC3-CTR cell proliferation was done using quantitative PCR (qPCR) analysis targeting the expression profiles of two PCa housekeeping genes, TATA-binding protein (TBP) and hypoxanthine phosphoribosyltransferase 1 (HPRT1) encoding genes. The excessive proliferation of PC3 cells in vivo was detected with both qPCR assays. Expression levels of one non­coding gene, prostate cancer associated 3 gene (pca3), and two other genes encoding transient receptor potential ion channel Melastatin 8 (trpm8) and prostate-specific membrane antigen (psma), showed a significantly enhanced aggressiveness of PC3-CTR cells in vivo. The model established in the present study provides an improved in vivo model for the diagnosis of PCas efficiently. This PCa cell xenograft model can also serve as a tool for high throughput anti-PCa drug screening in therapeutic treatments.

The effect of surface tension on steadily translating bubbles in an unbounded Hele-Shaw cell.

New numerical solutions to the so-called selection problem for one and two steadily translating bubbles in an unbounded Hele-Shaw cell are presented. Our approach relies on conformal mapping which, for the two-bubble problem, involves the Schottky-Klein prime function associated with an annulus. We show that a countably infinite number of solutions exist for each fixed value of dimensionless surface tension, with the bubble shapes becoming more exotic as the solution branch number increases. Our numerical results suggest that a single solution is selected in the limit that surface tension vanishes, with the scaling between the bubble velocity and surface tension being different to the well-studied problems for a bubble or a finger propagating in a channel geometry.

Experimental inoculation of Louisiana red swamp crayfish Procambarus clarkii with white spot syndrome virus (WSSV).

The red swamp crayfish Procambarus clarkii represents an important aquaculture species responsible for over half of all commercial aquaculture profits in Louisiana, USA. White spot syndrome virus (WSSV) is highly pathogenic in crustacean species and induces mass mortality in aquaculture operations worldwide. Natural outbreaks of WSSV occur yearly in cultured populations of crayfish in Louisiana. The goal of this study was to better understand the infectivity of WSSV in P. clarkii, by determining the minimum lethal dose necessary to initiate infection and to measure the resulting cumulative mortality following infection with different doses. A real time quantitative PCR (qPCR) method was used to detect WSSV in DNA extracted from gill tissue to ensure P. clarkii study populations were WSSV-free before the start of trials. Viable viral particles were isolated from naturally infected P. clarkii gill tissue and quantified using a novel digital PCR approach. Three infectivity trials were performed, and WSSV inocula were created by serial dilution, generating 5 treatments per trial. Five crayfish (weighing ~25 g) per dilution per trial received viral inoculations. Mortality was monitored daily for the duration of the trial in order to construct a median lethal dose (LD50) curve, and probit regression analysis was used to determine LD50 concentrations of viral particles. Knowledge of the infectivity of WSSV in native crayfish populations is of critical importance to the management of the commercial crayfish aquaculture industry in Louisiana. This is the first study to investigate the infectivity and to determine the LD50 of the Louisiana strain of WSSV in native crayfish.

Physiological management of dietary deficiency in n-3 fatty acids by spawning Gulf killifish (Fundulus grandis).

Lipid dynamics of spawning fish are critical to the production of viable embryos and larvae. The present study utilized manipulation of dietary fatty acid (FA) profiles to examine the ability of spawning Gulf killifish (Fundulus grandis) to mobilize critical lipid components from somatic reserves or synthesize long-chain polyunsaturated FAs (LC-PUFAs) de novo from shorter-chain C18 precursors. An egg and multi-tissue evaluation of changes in FA concentrations across time after fish were switched from LC-PUFA-rich to LC-PUFA-deficient experimental diets was employed. The two experimental diets contained lipid sources which differed drastically in n-3 C18 FA content but had similar levels of n-6 C18 FAs. Discrete effects of dietary n-3 FAs can be analyzed because n-3 and n-6 represent distinct metabolic families which cannot be exchanged in vivo. Results indicate that a combination of mobilization and de novo synthesis is likely utilized to maintain physiologically required FA levels in critical tissues and embryos. Mobilization was supported by decreases in LC-PUFAs in somatic tissues and decreases in intraperitoneal fat content and liver mass. Evidence for biosynthesis was provided by a higher level of n-3 LC-PUFAs in the liver and ova of fish fed diets containing n-3 C18 precursors versus those fed diets with low levels of precursor FAs. The characteristic physiological plasticity of Gulf killifish is exemplified in the nutritional domain by its management of dietary FA deficiency.

Diet-induced fatty acid variation in critical tissues of a spawning estuarine fish and consequences for larval fitness.

Freshwater and marine fishes exhibit a dichotomy in biosynthesis of long-chain polyunsaturated fatty acids (LC-PUFAs) from shorter-chain precursors. Marine species generally lack this ability due to deficiencies in requisite desaturase or elongase enzymes. Gulf killifish Fundulus grandis is a euryhaline cyprinodont whose fatty acid (FA) dynamics have not been previously examined. We utilized experimental variations in exogenous FAs available to spawning F. grandis in order to construct a multitissue evaluation of FA allocation and quantify effects on reproductive output and offspring fitness. No significant decrease in fecundity occurred in animals consuming low levels of LC-PUFAs, although embryo viability rates were affected. Maternal dietary FA variation did produce differences in starvation tolerance, hypoosmoregulatory ability, and acute thermal stress tolerance for larvae. These variations occurred at elevated salinities and temperatures seldom if ever encountered by F. grandis in culture or natural environments and are unlikely to affect larval survival in these settings. FA composition of eggs and tissues from spawning females suggested potential biosynthesis of LC-PUFAs from shorter-chain precursors in F. grandis. Results of this study suggest that F. grandis possesses physiological mechanisms allowing maintenance of reproductive function when subjected to dietary deficiencies in FAs generally considered essential for marine fishes.

Multiple steadily translating bubbles in a Hele-Shaw channel.

Analytical solutions are constructed for an assembly of any finite number of bubbles in steady motion in a Hele-Shaw channel. The solutions are given in the form of a conformal mapping from a bounded multiply connected circular domain to the flow region exterior to the bubbles. The mapping is written as the sum of two analytic functions-corresponding to the complex potentials in the laboratory and co-moving frames-that map the circular domain onto respective degenerate polygonal domains. These functions are obtained using the generalized Schwarz-Christoffel formula for multiply connected domains in terms of the Schottky-Klein prime function. Our solutions are very general in that no symmetry assumption concerning the geometrical disposition of the bubbles is made. Several examples for various bubble configurations are discussed.

Dispersant and salinity effects on weathering and acute toxicity of South Louisiana crude oil.

Chemical dispersants are an important technology in the remediation of oil spills in the aquatic environment, facilitating degradation of crude oil and salinity is an important factor in dispersant effectiveness. The aim of the present study was to explore the role of salinity on the degradation chemistry of crude oil polycyclic aromatic hydrocarbons (PAHs) and acute toxicity of the water accommodated fraction (WAF) of the dispersant COREXIT 9500A and chemically dispersed crude oil on a common estuarine fish. Laboratory microcosms were designed at salinities of 4 parts per thousand (ppt), 12 ppt, or 18 ppt and spiked with crude oil, COREXIT 9500A, or a combined exposure to crude oil and COREXIT and allowed to biodegrade for 1 wk, 4 wk, and 16 wk. The WAF was harvested for analytical PAH analysis and acute toxicity testing in juvenile Fundulus grandis. Compared with undispersed oil, COREXIT exponentially increased the PAH concentrations in the WAF for up to 16 wk; hopane-normalized concentrations indicated that biodegradation was slowed for the first 4 wk. Dispersed crude oil and COREXIT were acutely toxic following 1 wk of biodegradation with no correlation between PAH concentrations and crude oil WAF mortality. Both dispersant and dispersant oil mixtures remained toxic for at least 4 wk at the lowest salinity tested, suggesting increased sensitivity or reduced biodegradation of toxic components in low-saline environments. At the lowest salinity, oil dispersed with COREXIT was more toxic than either the COREXIT alone or oil alone, even after 16 wk of biodegradation.

Physiological biomarkers of hypoxic stress in red swamp crayfish Procambarus clarkii from field and laboratory experiments.

The crayfish industry in Louisiana is the largest in the United States, with crayfish frequently harvested from waters that experience episodic or chronic hypoxia (dissolved oxygen [DO]≤ 2 mg/l). We examined physiological biomarkers (hemolymph lactate, glucose, and protein concentrations) of hypoxic stress in the red swamp crayfish Procambarus clarkii from chronically hypoxic natural habitats and laboratory hypoxia experiments. P. clarkii from normoxic and hypoxic areas in the Atchafalaya River Basin were sampled monthly from April to July 2010. Laboratory experiments subjected P. clarkii to severe hypoxia (1 mg/l DO), moderate hypoxia (2 mg/l DO), or normoxic conditions (control: DO>7.5 mg/l) for 12, 24, and 48 h. P. clarkii from normoxic and hypoxic natural habitats did not display significantly different hemolymph lactate or glucose concentrations; however, mean hemolymph protein concentration was significantly lower in crayfish from hypoxic areas. P. clarkii exposed to severe hypoxia in laboratory experiments had significantly higher hemolymph lactate and glucose concentrations for all three exposure times, whereas large differences in protein concentrations were not observed. These results suggest that elevated hemolymph lactate and glucose concentrations are responses to acute hypoxia in P. clarkii, while differences in protein concentrations are the result of chronic hypoxic exposure.

Embryonic development and metabolic costs in Gulf killifish Fundulus grandis exposed to varying environmental salinities.

The Gulf killifish (Fundulus grandis) is a euryhaline fish found in coastal marsh along the entire of Gulf of Mexico and southern Atlantic of coast of the United States. The objective of this study was to investigate the effects of salinity on embryogenesis in the Gulf killifish. Four recirculation systems at salinities of 0.4, 7, 15, and 30 g/L were maintained at a static temperature with flow-through trays, containing embryos (n = 39) placed in triplicate into each system. Throughout embryogenesis, the rate of development, ammonia and urea excretion, and heart rate were monitored. Percent hatch was recorded, and morphological parameters were measured for larvae at hatch. As salinity was increased, the rate of embryogenesis decreased. Salinity significantly affected percent hatch with an 80.0% ± 2.6% for 7 g/L and 39.1 ± 4.3, 45.4 ± 4.5, and 36.3% ± 12.0% for 0.4, 15, and 30 g/L, respectively. Salinity and stage of development significantly affected production of ammonia and urea. As salinity increased, the dominate metabolite end product changed from urea to ammonia. However, the 15 g/L salinity treatment had the two highest levels of urea recorded. Heart rate was unaffected by salinity but increased throughout embryogenesis and remained constant once embryos reached stages where hatching has been recorded. While mean total length was not affected by salinity, embryos incubated in 30 g/L produced larvae with significantly thicker body depth at hatch. The 0.4, 7, and 15 g/L salinity treatments all had similar mean hours to hatch. The 30 g/L treatment resulted in a significantly longer mean time to hatch and smaller body cavity area at hatch.

Conversation effects on neural mechanisms underlying reaction time to visual events while viewing a driving scene: fMRI analysis and asynchrony model.

This neuroimaging study investigated the neural mechanisms of the effect of conversation on visual event detection during a driving-like scenario. The static load paradigm, established as predictive of visual reaction time in on-road driving, measured reaction times to visual events while subjects watched a real-world driving video. Behavioral testing with twenty-eight healthy volunteers determined the reaction time effects from overt and covert conversation tasks in this paradigm. Overt and covert conversation gave rise to longer visual event reaction times in the surrogate driving paradigm compared to just driving with no conversation, with negligible effect on miss rates. The covert conversation task was then undertaken by ten right-handed healthy adults in a 4-Tesla fMRI magnet. We identified a frontal-parietal network that maintained event detection performance during the conversation task while watching the driving video. Increased brain activations for conversation vs. no conversation during such simulated driving was found not only in language regions (Broca's and Wernicke's areas), but also specific regions in bilateral inferior frontal gyrus, bilateral anterior insula and orbitofrontal cortex, bilateral lateral prefrontal cortex (right middle frontal gyrus and left frontal eye field), supplementary motor cortex, anterior and posterior cingulate gyrus, right superior parietal lobe, right intraparietal sulcus, right precuneus, and right cuneus. We propose an Asynchrony Model in which the frontal regions have a top-down influence on the synchrony of neural processes within the superior parietal lobe and extrastriate visual cortex that in turn modulate the reaction time to visual events during conversation while driving.

Cognitive avionics and watching spaceflight crews think: generation-after-next research tools in functional neuroimaging.

Confinement and isolation have always confounded the extraordinary endeavor of human spaceflight. Psychosocial health is at the forefront in considering risk factors that imperil missions of 1- to 2-yr duration. Current crewmember selection metrics restricted to behavioral observation by definition observe rather than prevent performance degradation and are thus inadequate when preflight training cannot simulate an entire journey. Nascent techniques to monitor functional and task-related cortical neural activity show promise and can be extended to include whole-brain monitoring. Watching spaceflight crews think can reveal the efficiency of training procedures. Moreover, observing subcortical emotion centers may provide early detection of developing neuropsychiatric disorders. The non-invasive functional neuroimaging modalities electroencephalography (EEG), magnetoencephalography (MEG), magnetic resonance imaging (MRI), and near-infrared spectroscopy (NIRS), and highlights of how they may be engineered for spacecraft are detailed. Preflight and in-flight applications to crewmember behavioral health from current generation, next generation, and generation-after-next neuroscience research studies are also described. The emphasis is on preventing the onset of neuropsychiatric dysfunctions, thus reducing the risk of mission failure due to human error.