Advancing aquaculture: Production of xenogenic catfish by transplanting blue catfish (Ictalurus furcatus) and channel catfish (I. punctatus) stem cells into white catfish (Ameiurus catus) triploid fry.

Xenogenesis has been recognized as a prospective method for producing channel catfish, Ictalurus punctatus ♀ × blue catfish, I. furcatus ♂ hybrids. The xenogenesis procedure can be achieved by transplanting undifferentiated stem cells derived from a donor fish into a sterile recipient. Xenogenesis for hybrid catfish embryo production has been accomplished using triploid channel catfish as a surrogate. However, having a surrogate species with a shorter maturation period, like white catfish (Ameiurus catus), would result in reduced feed costs, labor costs, and smaller body size requirements, making it a more suitable species for commercial applications where space is limited, and as a model species. Hence, the present study was conducted to assess the effectiveness of triploid white catfish as a surrogate species to transplant blue catfish stem cells (BSCs) and channel catfish stem cells (CSCs). Triploid white catfish fry were injected with either BSCs or CSCs labeled with PKH 26 fluorescence dye from 0 to 12 days post hatch (DPH). No significant differences in weight and length of fry were detected among BSCs and CSCs injection times (0 to 12 DPH) when fry were sampled at 45 and 90 DPH (P > 0.05). The highest survival was reported when fry were injected between 4.0 to 5.5 DPH (≥ 81.2%). At 45 and 90 DPH, cell and cluster area increased for recipients injected from 0 to 5.2 DPH, and the highest cluster area values were reported between 4.0 to 5.2 DPH. Thereafter, fluorescent cell and cluster area in the host declined with no further decrease after 10 DPH. At 45 DPH, the highest percentage of xenogens were detected when fry were injected with BSCs between 4.0 to 5.0 and CSCs between 3.0 to 5.0 DPH. At 90 DPH, the highest number of xenogens were detected from 4.0 to 6.0 DPH when injected with either BSCs or CSCs. The current study demonstrated the suitability of white catfish as a surrogate species when BSCs and CSCs were transplanted into triploid white catfish between 4.0 to 6.0 DPH (27.4 ± 0.4°C). Overall, these findings allow enhanced efficiency of commercializing xenogenic catfish carrying gametes of either blue catfish or channel catfish.

Growth, survivorship, and predator avoidance capability of larval shortnose sturgeon (Acipenser brevirostrum) in response to delayed feeding.

Larval shortnose sturgeon, reared at 17°C, were subjected to delayed feeding treatments of 0, 5, 10, 15, 18, and 23 days post-yolk absorption to examine effects of food deprivation on growth, survival, swimming activity, and escape capabilities. Starvation affected growth and survival but despite degree of starvation, larvae were able to resume growth and experience high survivorship following feeding. Specific growth rate based on larval dry weight for the period directly following first feeding was highest for the day 15 and 18 delayed feeding treatments. There were no differences in survival between the 0 and 5 day treatments, however survival was reduced to 71.2%, 45.4%, and 28.8% for 10, 15, and 18 day delayed feeding treatments, respectively. Shortnose sturgeon had a point-of-no-return (PNR; 55.7% initiated feeding) at ~19 days (or 42 days post-fertilization) following the full absorption of yolk. Mean percent swimming activity and swimming speeds showed an interaction between delayed feeding treatment and larval age, such that no differences were detected at 1 and 6 days post-yolk absorption, while these swimming behaviors generally increased or spiked as feeding was delayed for 10, 15, and 18 days post-yolk absorption. At 23 days post-yolk absorption, only swimming speed increased for larvae that were denied food for 18 days. While there was an interaction between delayed feeding treatments and age for proportion of larvae exhibiting an escape response, generally, larvae from all feeding treatments exhibited a positive escape response. There were also interactions between delayed feeding treatments and age post-yolk absorption for mean and maximum escape speeds, such that less aggressive escape responses were typically detected the longer larvae were denied food. Our research suggests that larval shortnose sturgeon increase physical activity during periods of starvation to find a food patch while remaining vigilant but maybe not as capable to defend against a predatory attack as fed individuals.

Linking environmental factors with reflex action mortality predictors, physiological stress, and post-release movement behaviour to evaluate the response of white sturgeon (Acipenser transmontanus Richardson, 1836) to catch-and-release angling.

White sturgeon are the largest freshwater fish in North America and are the focus of an intense catch-and-release (C&R) fishery; the effects are largely unknown. We assessed the effect of fight and handling time, water temperature, river discharge rate, and fish size on physiological and reflex impairment responses of wild white sturgeon to angling. Sixty of these fish were tagged with acoustic transmitters to assess survival and post-release behaviour. Survival was high (100%). Water temperature and discharge influenced post-capture blood physiology. Specifically, lactate, chloride, and cortisol concentrations were elevated in individuals fought longer, and captured at higher water temperatures and river discharge. Cortisol was affected by fish size, with lower concentrations found in larger individuals. Only lactate and chloride were positively related to reflex impairment scores. Post-release movements were correlated with physiological state, fight characteristics and the environment. Specifically, higher blood lactate and chloride and those with longer fight times moved shorter distances after release. Contrastingly, higher levels of circulating glucose and potassium, as well as larger fish captured during periods of high discharge moved longer distances. Sturgeon tended to move shorter distances and at slower rates when reflex impairment was high, although reflex impairment in general did not explain a significant proportion of the variance in any movement metric. Our results show intriguing variance in the physiological and behavioural response of individual white sturgeon to C&R angling, with some degree of environmental dependence, and highlights the importance of understanding drivers of such variation when managing fisheries.

Frizzled 4 regulates ventral blood vessel remodeling in the zebrafish retina.

BACKGROUND: Familial exudative vitreoretinopathy (FEVR) is a rare congenital disorder characterized by a lack of blood vessel growth to the periphery of the retina with secondary fibrovascular proliferation at the vascular-avascular junction. These structurally abnormal vessels cause leakage and hemorrhage, while the fibroproliferative scarring results in retinal dragging, detachment and blindness. Mutations in the FZD4 gene represent one of the most common causes of FEVR. METHODS: A loss of function mutation resulting from a 10-nucleotide insertion into exon 1 of the zebrafish fzd4 gene was generated using transcription activator-like effector nucleases (TALENs). Structural and functional integrity of the retinal vasculature was examined by fluorescent microscopy and optokinetic responses. RESULTS: Zebrafish retinal vasculature is asymmetrically distributed along the dorsoventral axis, with active vascular remodeling on the ventral surface of the retina throughout development. fzd4 mutants exhibit disorganized ventral retinal vasculature with discernable tubular fusion by week 8 of development. Furthermore, fzd4 mutants have impaired optokinetic responses requiring increased illumination. CONCLUSION: We have generated a visually impaired zebrafish FEVR model exhibiting abnormal retinal vasculature. These fish provide a tractable system for studying vascular biology in retinovascular disorders, and demonstrate the feasibility of using zebrafish for evaluating future FEVR genes identified in humans.

Swimming kinematics and temperature effects on spermatozoa from wild and captive shortnose sturgeon (Acipenser brevirostrum).

Computer-assisted semen analysis (CASA) and cluster analysis were used to compare spermatozoa swimming kinematics and milt quality between wild and captive shortnose sturgeon (Acipenser brevirostrum). Milt samples from 27 shortnose sturgeon were collected in May 2016 and June 2017. Of these, 19 were wild caught in the Saint John River, New Brunswick, Canada, and eight were from a captive population at the Mactaquac Biodiversity facility. The following kinematic variables were measured immediately following sperm activation (˜5 s), at 30, 60, and 180 s post-activation; average path velocity (VAP); straight-line velocity (VSL); curvilinear velocity (VCL); amplitude of lateral head displacement (ALH); beat cross frequency (BCF); straightness (STR); linearity (LIN); wobble (WOB); percent motility (MOT). Analyses were conducted at 7, 10, and 14 °C to determine potential effects of temperature on kinematics. Principal components analysis (PCA) of original kinematic variables yielded two main components, a speed/wobble component along with a movement pattern component. Hierarchical cluster analysis (HCPC) indicated there were distinct subpopulations, with composition of clusters the result of fish source (wild-caught or captive). Wild-caught fish had greater sperm densities (P = 0.0064) and sperm swimming speeds compared to captive fish (P < 0.05). Temperature had a significant effect only on captive spermatozoa, and this result was not consistent between time periods. There was no effect of hormonal manipulation on spermatozoa motility kinematics. Results indicate there are significant differences in measures of milt quality between wild and captive shortnose sturgeon, indicating an effect of rearing condition on reproductive potential, which may affect fertilization success.

Discriminating Sex in Zebrafish (Danio rerio) Using Geometric Morphometrics.

Zebrafish (Danio rerio) adults are viewed as sexually dimorphic. However, current approaches to sex discrimination rely mainly on subjective assessment of color patterns and body structures. Here, we explore how geometric morphometrics allow for quantitative sex discrimination based on overall body geometry of adult zebrafish (aged 12-24 months). Ten homologous landmarks were placed on the left lateral view of adult zebrafish and transformed through Procrustes superimposition before being analyzed with canonical variate analysis. We compared two models to distinguish between sexes. The first model consisted of landmarks that included the abdominal region and the second model did not. Males and females clearly diverged along a single canonical variate, and jackknife testing reinforced the strength of the sorting algorithm with 100% correct assignment of sex for both models. Analysis of body geometry demonstrated that males have a longer caudal peduncle, a more streamlined ventral region, and slightly more inferior placement of eyes than females. Based on these results we developed a logistic regression equation using the ratio of ventral caudal peduncle length to standard length to provide researchers a reliable and objective method for sex discrimination in zebrafish.

Oceanic Distribution, Behaviour, and a Winter Aggregation Area of Adult Atlantic Sturgeon, Acipenser oxyrinchus oxyrinchus, in the Bay of Fundy, Canada.

Seasonal distribution of adult Atlantic sturgeon was examined using pop-up satellite archival tags (PSATs) and ultrasonic transmitters deployed in the Saint John River, New Brunswick, Canada. Seven MK10 PSATs programmed for release in June 2012 and seven MiniPAT PSATs programmed for release in February and April 2013 were deployed in August 2011 and 2012, respectively. Eleven of 14 PSATs surfaced and transmitted depth and temperature data archived for the duration of their deployment (121-302 days). Among these eleven PSATs, five were recovered and 15-sec archival data was downloaded. Following exit from the Saint John River in the fall, tagged fish occupied a mean monthly depth of 76.3-81.6 m at temperatures as low as 4.9˚C throughout the winter before returning to shallower areas in the spring. The majority of ultrasonic detections occurred in the Bay of Fundy, but fish were detected as far as Riviere Saint-Jean, Quebec, approximately 1500 km from the Bay of Fundy (representing long-distance migratory rates of up to 44 km/day). All PSATs were first detected in the Bay of Fundy. Tags that released in February and April were found 5-21 km offshore of the Saint John Harbour, while tags that released in June were first detected in near shore areas throughout the Bay of Fundy. The substrate at winter tag release locations (estimated from backward numerical particle-tracking experiments) consisted primarily of moraines and postglacial mud substrate with low backscatter strength, indicative of soft or smooth seabed. Based on the proximity of winter tag release locations, the consistent depths observed between fish, and previous research, it is suspected that a winter aggregation exists in the Bay of Fundy. This study expands the understanding of the marine distribution and range of Atlantic sturgeon on the east coast of Canada.

Seminal plasma biochemistry and spermatozoa characteristics of Atlantic cod (Gadus morhua L.) of wild and cultivated origin.

Our objectives were to compare spermatozoa activity, morphology, and seminal plasma (SP) biochemistry between wild and cultivated Atlantic cod (Gadus morhua). Swimming velocities of wild cod spermatozoa were significantly faster than those of cultivated males. Wild males had a significantly larger spermatozoa head area, perimeter, and length, while cultivated males had more circular heads. Total monounsaturated fatty acids and the ratio of n-3/n-6 were significantly higher in sperm from wild males, while total n-6 from cultivated males was significantly higher than the wild males. Significantly higher concentrations of the fatty acids C14:0, C16:1n-7, C18:4n-3, C20:1n-11, C20:1n-9, C20:4n-3, C22:1n-11, and C22:6n-3 were observed in wild males, while significantly higher concentrations of C18:2n-6, C20:2n-6, and C22:5n-3 occurred in cultivated males. Osmolality, protein concentration, lactate dehydrogenase and superoxide dismutase activity of SP of wild males were significantly higher than the cultivated males. Antioxidant capacity of SP was significantly higher in cultivated males, while pH and anti-trypsin did not differ between fish origins. Four bands of anti-trypsin activity and nine protein bands were detected in SP. Performing a discriminant function analysis, on morphology and fatty acid data showed significant discrimination between wild and cultivated fish. Results are relevant to breeding programs and aquaculture development.