Triploid salmonids have been shown to underperform in suboptimal environments. It is thought this might be due to having larger cells to accommodate the increased number of chromosomes and therefore effects on aerobic metabolism from having smaller cellular surface area to volume ratios. The goal of this study was to examine the aerobic metabolism of diploid and triploid white sturgeon (Acipenser transmontanus) in ambient (18 °C) and elevated water temperatures (24 °C). Resting and maximum metabolic rates, recovery time from exhaustive exercise, and surface area to volume ratios of erythrocytes and their nuclei in diploid and triploid sturgeon were evaluated. Triploid sturgeon had a reduced aerobic scope and hematological response (hematocrit and hemoglobin) to exhaustive exercise. A reduced surface area to volume ratio of erythrocytes in triploid sturgeon provides evidence that cellular surface area could be one mechanism limiting aerobic metabolism in triploid fishes. A lower aerobic scope found in triploid sturgeon may impact reproductive and somatic growth, yet more research is needed to determine implications for management decisions on farms and hatcheries.

Previous studies have demonstrated reduced performance in triploid fish when reared under suboptimal conditions, which may be the result of a higher susceptibility to stressors when compared to diploids. The goal of this project was to investigate differences in the capacity of diploid (8 N) and triploid (12 N) white sturgeon, Acipenser transmontanus, to respond to both warm acclimation (6-weeks of acclimation to either 18 or 22 °C) and a subsequent acute stress (10-min low water stress). Following the 6-week acclimation, fish were sampled either before or following an acute low water stress. Bioindices of the primary and secondary stress response, hematology and cellular metabolic status were measured. We also sought to determine if time to peak cortisol levels were similar between diploid and triploid sturgeon after exposure to a severe acute stressor (netting stress). While both ploidies had similar primary and secondary responses to acute stress, both with and without warm acclimation, warm acclimation impacted the ability of diploid and triploid white sturgeon to mount a typical stress response to an acute stressor. In response to warm acclimation, triploids exhibited little change in branchial lactate dehydrogenase activity, while diploids increased activity. After exposure to an acute water reduction stress, diploids increased citrate synthase activity, yet triploids showed a decrease in activity. Differences in metabolic enzyme activity in response to warm acclimation and acute stress suggest triploid white sturgeon may have a reduced cellular metabolic capacity under chronic and acute stress, which may impact performance of triploid sturgeon in suboptimal conditions.

Previous studies suggest fish with additional copies of their genome (polyploids) underperform in suboptimal conditions and may be more susceptible to stress and disease. The objective of this study was to determine the role ploidy plays in the physiological response of white sturgeon to chronically elevated water temperatures. White sturgeon of two ploidies (8 N and 10 N) were acclimated to ambient (18 °C) and warm (22 °C) water. Bioindices of stress (plasma cortisol, glucose and lactate, total erythrocyte count, hematocrit, hemoglobin, mean erythrocyte volume, mean erythrocyte hemoglobin, and mean erythrocyte hemoglobin concentration), immunity (respiratory burst, plasma lysozyme, and total leukocyte count), and cellular metabolic capacity (lactate dehydrogenase and citrate synthase activity) were measured before and after a 6-week acclimation period. Both ploidies appear comparable in their constitutive immune and stress parameters and respond similarly to warming. Hematological indices suggest 8 N and 10 N sturgeon are similar in oxygen carrying capacity; however, differences in enzyme activity between ploidies indicate that 10 N sturgeon may have a lower cellular aerobic capacity. Our results have implications for the screening and management of ploidy on white sturgeon farms and hatcheries, as the differences between ploidies may affect 10 N sturgeon performance at elevated water temperatures. Further research is needed to elucidate the differences in inducible stress and immune responses and metabolism of white sturgeon of different ploidies.

Sturgeon and paddlefishes (Acipenseriformes) are cultured worldwide for commercial and conservation purposes. Over the past decade, we have learned many sturgeon culture programs experience spontaneous autopolyploidy, or the generation of triploidy without management intervention. Spontaneous autopolyploidy has been shown to negatively impact the physiology, reproductive development and fertility of sturgeon. Given these concerns, it is necessary to understand how it arises, how it affects production, and how it can be managed. The goal of this review is to describe the incidence of spontaneous autopolyploidy in sturgeon relative to other fishes, explain how spontaneous autopolyploids are formed, and report what is known about how triploidy and intermediate ploidy effect sturgeon reproductive development and physiology. We then describe four common methods for detecting spontaneous autopolyploids and their intermediate ploidy progeny and end with recommendations for reducing the incidence of spontaneous autopolyploidy based on the results of recent research. Although spontaneous autopolyploidy may never be completely eliminated from sturgeon culture, following these recommendations may significantly decrease its incidence.

Previous studies have provided evidence of a reduced aerobic metabolic capacity, both at the cellular (metabolic enzyme activity) and the whole organism (aerobic scope) level in juvenile triploid white sturgeon, compared to diploid siblings. The downstream costs of this reduced metabolic capacity are still unclear, yet a lower aerobic scope suggests triploid white sturgeon likely have less energy to allocate to biological processes like growth and development. We conducted a 15-week growth trial to assess energy allocation to somatic growth in 2-month-old diploid and triploid white sturgeon. Spontaneous swimming activity, hepatosomatic index, condition factor, and deformities were also measured throughout the growth trial as indices of energy allocation to activity and fish condition. In general, our results indicate that triploid white sturgeon may have less energy available for processes beyond basal maintenance. This could be linked to a reduced overall performance as evidenced by lower weights and more deformities when compared with their diploid counterparts. However, many indices were still mostly unaffected by triploidy (condition factor, hepatosomatic index, and swimming activity). Whether this lower growth performance seen in juvenile triploid white sturgeon continues through sub-adulthood, puberty and final maturation requires further long-term studies.

Using a system optimized for propagating human keratinocytes, culture of skin samples from white and green sturgeons generated epithelial cells capable of making cross-linked protein envelopes. Two distinct forms of TGM1-like mRNA were molecularly cloned from the cells of white sturgeon and detected in green sturgeon cells, accounting for their cellular envelope forming ability. The protein translated from each displayed a cluster of cysteine residues resembling the membrane anchorage region expressed in epidermal cells of teleosts and tetrapods. One of the two mRNA forms (called A) was present at considerably higher levels than the other (called B) in both species. Continuous lines of white sturgeon epidermal cells were established and characterized. Size measurements indicated that a substantial fraction of the cells became enlarged, appearing similar to squames in human epidermal keratinocyte cultures. The cultures also expressed CYP1A, a cytochrome P450 enzyme inducible by activation of aryl hydrocarbon receptor 2 in fish. The cells gradually improved in growth rate over a dozen passages while retaining envelope forming ability, TGM1 expression and CYP1A inducibility. These cell lines are thus potential models for studying evolution of fish epidermis leading to terrestrial adaptation and for testing sturgeon sensitivity to environmental stresses such as pollution.

The authors regret that the incorrect version of Figure 4 was included in the original paper.

Spontaneous autopolyploidy, a 1.5x increase in genome size resulting from second polar body retention after fertilization, has been documented in cultured sturgeons, with the proportion of spontaneous autopolyploid progeny ranging widely among maternal families. Sturgeon farms wish to reduce the number of spontaneous autopolyploids because their progeny, when crossed with a normal ploidy parent, exhibit intermediate ploidies, resulting in reproductive abnormalities. However, there is limited knowledge of the causes of the second polar body retention in sturgeon culture. In this study, we report the results of experiments performed from 2015 to 2019 aimed at identifying the sources of spontaneous autopolyploidy in white sturgeon (Acipenser transmontanus) culture. In collaboration with several sturgeon farms, we tested whether post-ovulatory ageing, mechanical shock during egg de-adhesion, and the combined effects of both factors increased spontaneous autopolyploidy. To test the effect of post-ovulatory ageing, eggs were collected from females and either fertilized at the industry normative time (2–5 h post-oviposition) or stored in ovarian fluid at 15 °C for 6–8 h before fertilization. To test the effect of mechanical shock, eggs were collected, fertilized 2–5 h post-oviposition and exposed to either gentle or vigorous mixing during the 60 min de-adhesion treatment. Results from this work reveal that post-ovulatory ageing does increase the incidence of spontaneous autopolyploidy in some females, but overall the proportions produced were low (range 0–15%). Proportions of spontaneous autopolyploids in eggs exposed to vigorous mixing were also variable (1–92%) but significantly higher in 75% of the females when compared to their respective controls or gentle mixing treatments, indicating that mechanical shock during egg de-adhesion is likely the primary cause of spontaneous autopolyploidy. To our knowledge this is the first study to document mechanical shock to eggs during de-adhesion as a cause of abnormal ploidy in cultured fishes. We observed high variability in egg quality among females and a significant relationship between embryo mortality and the incidence of spontaneous autopolyploidy when eggs were exposed to mechanical shock. Repeated spawning of a female that produced a high proportion of spontaneous autopolyploids provided preliminary evidence that genetic background may influence the likelihood that a female's eggs will be prone to second polar body retention when subjected to mechanical shock. Although spontaneous autopolyploidy in sturgeon culture will likely never be eliminated entirely, we provide practical recommendations to sturgeon producers to reduce its incidence in a production setting.

Release of sturgeon with abnormal ploidy into the wild may result in reduced fitness due to lowered fertility in the F₂ and subsequent generations. Further, there is evidence that ploidy affects reproductive development and caviar yield. Therefore, the ability to accurately characterize the ploidy of white sturgeon (Acipenser transmontanus) is essential for both commercial and conservation aquaculture. This study compares nuclear volume and whole erythrocyte long-axis lengths obtained using Coulter counter and blood smears, respectively, from captive white sturgeon from populations originating in California and Idaho. We examine which method provides the most accurate, time efficient and cost-effective characterization of ploidy in this species. Results from Coulter counter and blood smears were compared to results from flow cytometry, the gold standard for genome size analysis. Previous work suggests that blood smears can distinguish between 8N (diploid) and 12N (triploid) sturgeon, but further analysis is required to see if this method can also be used to identify 10N fish and to provide robust evidence of its utility in 8N and 12N fish across populations. In this study, we demonstrated that the Coulter counter had 100% agreement with flow cytometry in ploidy assignment, while blood smears vary in their accuracy based on population. Blood smears showed a high degree of overlap in erythrocyte long-axis length between 8N and 10N individuals as well as some overlap between 10N and 12N individuals in the California fish, and a high degree of overlap between 8N and 12N individuals in the Idaho fish. Although blood smears are time-intensive and vary in their ploidy assignment accuracy, they are a low-cost technique and as such may have some utility for caviar farms attempting to identify 12N individuals in a small number of broodstock. By comparing the accuracy, efficiency and cost of these three methods, sturgeon farmers and conservation hatcheries will be able to choose the best method for their needs in determining the ploidy of their fish. We determined that Coulter counter is equally accurate to flow cytometry and is also the most time efficient method for ploidy determination in white sturgeon.

With the increasing use of triploid fish for sport fisheries management, biologists, researchers, and managers working in field environments need practical methods to determine ploidy. Cytological methods (e.g., flow cytometry, Coulter counter) using erythrocytes are the most common for ploidy determination in fishes. However, collecting and storing erythrocytes can be logistically challenging during field work, and donor fish need to be alive or freshly killed. With rapid advances in molecular genetics, biologists, researchers, and managers may be unaware of molecular approaches for ploidy determination that could alleviate the difficulties associated with cytological methods and allow ploidy determination from archived samples (e.g., fin clips, scales, otoliths). In this study, we analyzed the agreement between molecular-based (using fin tissue) and Coulter counter-based (using blood) ploidy determinations for Walleyes Stizostedion vitreum—the first assessment of concordance between molecular and cytological methods for determining ploidy in the family Percidae. We found that agreement between these two methods was >98%. The high degree of agreement and greater ease of collecting and storing samples for molecular-based approaches relative to the traditional cytological ones support the utility of molecular methods for ploidy determination.