Aspects of early development in the Adriatic sturgeon Acipenser naccarii

This study investigates the development of the sensory-cutaneous apparatus and digestive tract of Adriatic sturgeon (Acipenser naccarii). Light and electron microscopy observations were carried out on various developmental stages, from hatching to 180 days old.
At hatching, sturgeon pre-larvae exhibit differentiation of olfactory sensory cells. By day 4 taste buds are differentiated on lips and barbels. At 12 days after hatching, the larvae are equipped with an extensive ventral cephalic sensory apparatus. Electrosensory organs are arranged in regular lines in the rostral ventral region, and taste receptors are organised in parallel rows on and inside the lips as well as on the external side of the barbel. Ventral free neuromasts are positioned in rostral grooves. The retina is completely differentiated in each stratum. At this stage, larvae show canine-like teeth on lips and pharynx, and the specific mucosae of the different digestive regions are differentiated. By day 36 the canine-like teeth are located exclusively on the tongue and roof of the buccal cavity, and the mouth is protrusible. At 180 days differentiation is still not completed, and although teeth have disappeared from the palate, they still persist along a central line on the tongue.     

Swimming alters responses to hypoxia in the Adriatic sturgeon Acipenser naccarii

When the Adriatic sturgeon Acipenser naccarii was exposed to progressive hypoxia under static conditions, it exhibited a linear decline in O₂ uptake, behaving as an 'oxyconformer'. When, however, it was allowed to swim at a low sustained speed, it could regulate O₂ uptake down to a mean ± s . e . critical ( P _crit) of 4·9 ± 0·5 kPa ( n = 6). At moderate levels of hypoxia, static fish exhibited significant reductions in arterial blood O₂ content, and increases in plasma lactate, which were not observed in swimming animals.     

Cardiovascular responses to hypoxia in the Adriatic sturgeon (Acipenser naccarii)

The in vivo cardiovascular responses to hypoxia, and the intrinsic functional characteristics of the heart in vitro , were determined, and compared, in the Adriatic sturgeon (Acipenser naccarii). During exposure to hypoxia in vivo , blood oxygen content (Cao₂) declined as water 0₂ partial pressure (Pwo₂) was reduced, despite an increase in haematocrit. The main cardiovascular response was a reduction in dorsal aortic blood pressure, with a slight bradycardia, while cardiac output remained constant. Reduced oxygen content of the perfusate had significant inhibitory effects on the intrinsic performance of the heart in vitro , causing a reduction in the heart rate; a reduction in the sensitivity of responses to increased preload (Frank-Starling response), and a more rapid decline in power output and stroke volume when afterload was increased. Overall, the in vitro results suggest that hypoxia depresses the contractility of the heart (i.e. its inotropic responses). The reduction in dorsal aortic pressure in vivo may, therefore, counteract the depressive effects of hypoxia on heart contractility, and thereby avoid a hypoxic depression of cardiac output.     

Geometric morphometries in the characterisation of the cranial growth pattern of Adriatic sturgeon Acipenser naccarii

Geometric morphometries are a set of recently developed techniques in the study of biological forms. In this paper these techniques are used in the characterisation of shape changes of the cranial region during growth of the Adriatic sturgeon Acipenser naccarii. Species identification in sturgeons is sometimes difficult, biased by morphological plasticity, by individual variability, complicated by human intervention and manipulation, such as aquaculture, hybridisation, pollution and shape changes during growth (allometry). Characteristic changes in head morphology during subadult growth concern the relative shortening and the lateral stretching of the entire head region. These and other observed shape changes were never previously visualised nor reported in the literature. Geometric morphometries is proposed as a good candidate to became the universal methodology in the morphometrical analysis of sturgeons.     

Purification and characterization of the hemoglobin components of Adriatic sturgeon (Acipenser naccarii) blood

The oxygen binding properties of Acipenser naccarii hemoglobins have been investigated, and were found not to differ significantly from those shown by blood, intact erythrocytes and hemolysate in the presence of the physiological cofactors GTP and chloride ions. In particular, the oxygen equilibrium reveal a very low sensitivity of the hemolysate to the chloride ions and temperature while in the presence of organic phosphates their oxygen affinity decreased strongly. The electrophoretic analysis of the hemolysate of the sturgeon showed the presence of two hemoglobin components, each with a considerable globin multiplicity. The partial amino acid sequence of the a and P chain of the single hemoglobins was also established.     

Managing polyploidy in ex situ conservation genetics: the case of the critically endangered Adriatic sturgeon (Acipenser naccarii)

While the current expansion of conservation genetics enables to address more efficiently the management of threatened species, alternative methods for genetic relatedness data analysis in polyploid species are necessary. Within this framework, we present a standardized and simple protocol specifically designed for polyploid species that can facilitate management of genetic diversity, as exemplified by the ex situ conservation program for the tetraploid Adriatic sturgeon Acipenser naccarii. A critically endangered endemic species of the Adriatic Sea tributaries, its persistence is strictly linked to the ex situ conservation of a single captive broodstock currently decimated to about 25 individuals, which represents the last remaining population of Adriatic sturgeon of certain wild origin. The genetic variability of three F1 broodstocks available as future breeders was estimated based on mitochondrial and microsatellite information and compared with the variability of the parental generation. Genetic data showed that the F1 stocks have only retained part of the genetic variation present in the original stock due to the few parent pairs used as founders. This prompts for the urgent improvement of the current F1 stocks by incorporating new founders that better represent the genetic diversity available. Following parental allocation based on band sharing values, we set up a user-friendly tool for selection of candidate breeders according to relatedness between all possible parent-pairs that secures the use of non-related individuals. The approach developed here could also be applied to other endangered tetraploid sturgeon species overexploited for caviar production, particularly in regions lacking proper infrastructure and/or expertise.     

Influence of sampling conditions on blood chemistry values of Adriatic sturgeon Acipenser naccarii (Bonaparte, 1836)

Data on the blood chemistry of a chondrostean fish, the Adriatic sturgeon ( Acipenser naccarii ), are reported as measured with different sampling procedures, and as related to rearing conditions and age. Serum Cortisol, glucose, osmolality, Na⁺, Cl^-, Ca²⁺ and total protein concentrations were measured. Reference values for the blood chemistry of farmed sturgeon were measured on samples from resting undisturbed animals collected via a chronic indwelling catheter in the dorsal aorta that was implanted under anaesthesia. Following 24h recovery from catheterization, serum Cortisol, glucose and osmolality levels were 9.4 ng/ml, 58.8 mg/dl and 261.4 mOsm/kg, respectively. Furthermore, blood samples collected with the chronic indwelling catheters indicated that the surgical procedure of cannulation caused a stress response, with physiological changes that followed a pattern like that described in teleosts. Cortisol, glucose and osmolality were more sensitive to stress than the other variables measured. Sampling by cardiac puncture tended to be associated with elevated serum Cortisol levels in older, larger sturgeon, but not in young fish. Greater capture, confinement and handling stress in older, larger, sturgeon may have been responsible for this and other age-related differences in blood chemistry values measured following cardiac puncture. Within the same age class, both rearing conditions and temperature affected Cortisol, sodium and total protein concentrations significantly. Anaesthesia did not appear to reduce the degree of stress associated with cardiac puncture but altered serum ion concentrations.     

The influence of dietary fatty acid composition on the respiratory and cardiovascular physiology of Adriatic sturgeon (Acipenser naccarii): a review

This paper reviews evidence that the fatty acid composition of dietary lipids influences the respiratory and cardiovascular physiology of Adriatic sturgeon {Acipenser naccarii) and, thereby, their tolerance of the stress of hypoxia. Sturgeon fed a commercial diet enriched in fish oil (menhaden oil as 15% of dry feed weight), with an elevated content of highly unsaturated fatty acids of the co3 series (ω3 HUFA), had a significantly lower standard metabolic rate (SMR) and routine oxygen consumption (Mo₂) than those fed a diet enriched with the same quantity of hydrogenated coconut oil, with an elevated content of saturated fatty acids (SFA). Both groups grew equally well. As a result of this difference in aerobic metabolism, sturgeon fed the w3 HUFA and SFA responded differently when exposed to hypoxic challenges. Sturgeon fed w3 HUFA exhibited no significant reflex hyperventilation when exposed to mild, moderate or deep hypoxia (30 min at water 0₂ partial pressures of 10.8, 6.6 and 4.6 kPa, respectively), no hypoxic depression of spontaneous activity during 3h in mild hypoxia, and no depression of Mo₂ during 3h in moderate hypoxia, unlike sturgeon fed SFA. The diets also influenced the performance of isolated hearts in vitro. Hearts from fish fed o3 HUFA maintained maximum in vitro cardiac power output unchanged when oxygen supply was reduced (O₂ content from 2.3 to 0.7 vol.%), unlike hearts from sturgeon fed SFA. Overall, the results indicate that dietary fatty acid composition can influence tolerance of hypoxia in sturgeon, through effects on SMR. When compared to sturgeon fed SFA, those fed co3 HUFA had lower SMR and were more tolerant of hypoxia, with effects both on the whole animal and on the isolated heart.     

Regulation of calcium balance in the sturgeon Acipenser naccarii: a role for PTHrP

Calcium regulation in sturgeon is of special interest because they are a representative of the ancient fishes possessing mainly cartilaginous skeletons and a supposedly low calcium demand. The present study aimed to characterize the effect of a chronic absence of dietary calcium and the effect of parathyroid hormone-related protein (PTHrPA) (1-34) (7) on calcium balance in juvenile sturgeon (Acipenser naccarii). At rest, sturgeon juveniles are in net positive calcium balance, since whole body calcium uptake is significantly higher than efflux and calcium accumulates in the body. To study the importance of dietary calcium, the sturgeon were kept on a calcium-free diet for 8 wk. This manipulation impaired growth as measured by failure to gain weight or increase in length and indicates that dietary calcium is important for growth in sturgeon. An increased whole body calcium uptake partially compensated dietary calcium deficiency and was associated with increased gill chloride cell number in lamellae and filaments in parallel with increased gill Na(+)K(+)-ATPase activity. In addition, a single injection of piscine PTHrP(1-34) significantly increased whole body calcium uptake and decreased whole body calcium efflux. Administration of PTHrP significantly increased circulating plasma calcium 4-24 h postinjection. The increase in net calcium transport and increased plasma levels of calcium is consistent with the actions of a hypercalcemic factor. It would appear that the sturgeon rely on calcium for growth and tightly regulate calcium transport. The action in calcium balance is consistent with PTHrP acting as a hypercalcemic factor in sturgeon.     

Digestive enzymes during ontogeny of the sturgeon Acipenser naccarii: intestine and pancreas development

The present work examines the temporal appearance and degree of activity of digestive enzymes of pancreatic and intestinal origin (amylase as well as neutral and alkaline proteases) together with the evolution of the ontogenic development of the intestine and pancreas during the first month of free‐living (30 days post‐hatch–[dph]) of the sturgeon Acipenser naccarii. In addition, the influence of exogenous live feeding with Artemia salina on the detection of digestive enzymes was studied. Shown are that in the first life stages of the A. naccarii, from the time of fertilisation to the juvenile stage, the following events related to its digestive physiology occurred: digestive enzymes were detected in the embryo stage within the egg, presumably related to the hatching gland; in the free‐living embryo stage, opening of the mouth was at 10 dpf and protease and amylase digestive activities occurred due to an early differentiation of the pancreas and intestine (12–13 dpf). At this stage, digestive enzymatic activities are partly owing to the live feed offered before disappearance of the yolk reserves (14 dpf); the expulsion of the melanin plug and the appearance of a continuity of the digestion tract (16 dpf) together with the disappearance of the yolk reserves (17 dpf) mark the passage from the free‐living embryo phase to the juvenile stage; finally, from the first month after hatching in this sturgeon species, there is a stabilisation in digestive amylase and protease activities together with a fully developed digestive structure. The contribution of enzymes from prey in the detection of enzymatic activities determined in the fish was beyond doubt; therefore these exogenous enzymes must also have a certain physiological importance. Subsequent work will be needed to clarify the importance of digestive processes of the exogenous enzyme pool from live feed as well as to assess the possibility of shortening the weaning period.     

The influence of salinity on ionic concentrations and osmolarity of blood serum in lake sturgeon, Acipenser fulvescens

This study examined the extent to which lake sturgeon, Acipenser fulvescens, regulated serum ion concentrations and osmolarity in relation to ambient salinity. Twenty juvenile lake sturgeon, brood stock from the Moose River in the Hudson Bay drainage basin, were gradually transferred from freshwater to more saline waters at a rate of 2 day^-1. Blood was sampled from fish at 0, 10, 15 and 25 following 48 h acclimation periods. Concentrations of Na⁺, K⁺, Cl^-, Ca²⁺ and osmolarity were measured. Experimentation was terminated at 25 due to poor fish health resulting in two mortalities. Within freshwater, lake sturgeon blood serum displays similar osmotic and ionic composition to other North American species of sturgeon. Juvenile lake sturgeon do not appear capable of homeostasis above 15 nor are they tolerant to environments exceeding 25. Some evidence of greater tolerance to elevated salinities with increased size was noted. The ecological potential for resource exploitation in regions of elevated salinities is discussed.     

The acute temperature tolerance of green sturgeon (Acipenser medirostris) and the effect of environmental salinity

We investigated the effect of environmental salinity on the upper thermal tolerance of green sturgeon (Acipenser medirostris), a threatened species whose natural habitat is vulnerable to temperature and salinity variation as a result of global climate change. Freshwater (FW)-reared sturgeon were gradually acclimated to salinities representing FW, estuary water (EST), or San Francisco Bay water (BAY) at 18 degrees C, and their critical thermal maximum (CTMax) was measured by increasing temperature 0.3 degrees C/min until branchial ventilation ceased. CTMax was 34.2+/-0.09 degrees C in EST-acclimated fish, with FW- and BAY-acclimated fish CTMax at 33.7+/-0.08 and 33.7+/-0.1 degrees C, respectively. Despite the higher CTMax in EST-acclimated fish, FW-acclimated sturgeon ventilation rate reached a peak that was 2 degrees C higher than EST- and BAY-acclimated groups and had a greater range of temperatures within which they exhibited normal ventilatory function as assessed by Q(10) calculation. The osmoregulatory consequences of exposure to near-lethal temperatures were assessed by measuring plasma osmolality and hematocrit, as well as white muscle, brain, and heart tissue water contents. Hematocrit was increased following CTMax exposure, most likely owing to the elevated metabolic demands of temperature increase, and plasma osmolality was significantly increased in EST- and BAY-acclimated fish, which was likely the result of a greater osmotic gradient across the gill as metabolism increased. To our knowledge, this represents the first evidence for an effect of salinity on the upper thermal tolerance of sturgeon, as well as the first investigation of the osmoregulatory consequences of exposure to near-lethal temperatures. J. Exp. Zool. 309A:477-483, 2008. (c) 2008 Wiley-Liss, Inc.     

Cytochrome P-450 levels and EROD activity in the Adriatic sturgeon (Acipenser naccarii, Chondrostei) and its induction by β-Naphthoflavone

The cytochrome P-450 enzyme system was studied in juvenile Acipenser naccarii (317.6±77 g initial live weight) reared at 23°C and at a standard diet over a period of two month before using for experiments (final weight 546.3±148.8 g). While determining normal levels of cytochrome P-450 in liver tissue its induction was also studied under exposure to P-naphthoflavone (BNF; single dose at 80 mg/Kg body weight 52 hrs prior to sampling). Control fish showed total cytochrome P-450 levels in liver microsomes around 0.40±0.12 nmoles/mg proteins (n=12) while in BNF exposed fish the values increased significantly (0.55±0.11nmoles/mg proteins; n=12). Ethoxyresorufin-O-deethylase (EROD) in control fish was 0.015±0.02 nmoles/mg proteins/min (n=12); it was found to be NADPH dependent and inducible by BNF. Western Immunoblotting and enzyme-linked-immunosorbent (ELISA) analyses of liver microsomes from untreated and BNF-exposed fish, using an antibody to rat cytochrome P-450 1A1 isoform (P-450 1A1) (Nelson et al.,1996) suggested the presence of a constitutive and BNF-inducible cytochrome P-450 homologous to rat P-450 protein.
Measurement of cytochrome P-450-dependent EROD activity and P-450 1A1 expression levels in A. naccarii may be a useful parameter in monitoring environmental pollution.     

Ontogenesis of osmotic regulation in the striped mullet, Mugil cephalus L.

The osmoregulatory capabilities of juvenile striped mullet, Mugil cephalus L ., of three size-groups (20–29, 30–39 and 40–9 mm s.l.) were compared in a series of six salinities ranging from fresh water to full sea water. The two smaller size-groups were able to tolerate instantaneous transfer from the brackish water in which captured, to all salinities but fresh water, while the 40–69 mm group were tolerant of instantaneous transfer to all experimental salinities. At high environmental salinities, the osmotic regulatory capabilities improved with growth in size to an apparently definitive condition in fish of 40–69 mm s.l. The smallest of these individuals were estimated to be about 7.5 months old. The osmotic regulatory capability in waters of low salinity had reached a definitive state with the development of tolerance to such salinities. The ontogenetic pattern for Mugil cephalus is virtually identical to that of the Atlantic salmon, Salmo salar .     

Effect of starvation and refeeding on digestive enzyme activities in sturgeon (Acipenser naccarii) and trout (Oncorhynchus mykiss)

The digestive enzyme activities were determined in Adriatic sturgeon and rainbow trout during starvation and refeeding period. Overall, the digestive enzyme activities are affected in the same sense in both species. The protease and lipase activities were decreased later than amylase activity. Even after 1 month of starvation, both species would be prepared to digest protein and lipids in an effective way. After 72 days of starvation, the digestive machinery of the sturgeon and of the trout shows an altered capacity to digest macronutrients. The capacity to digest proteins and lipids, after 60 days of refeeding, begins to become re-established in sturgeon and trout. In contrast, in this period, the capacity to digest carbohydrates remains depressed in both species.