Post‐glacial history and introgression in Abies (Pinaceae) species of the Russian Far East inferred from both nuclear and cytoplasmic markers

Aim The main aim of the present study is to infer the post‐glacial history of Abies species from north‐east Asia and to test the hypotheses that coastal Abies populations suffered less from climatic fluctuations during Pleistocene glacial periods than their more continental counterparts, and that Sakhalin was a major area of introgression. Location Natural ranges of the fir species Abies nephrolepis, Abies sachalinensis and Abies holophylla in the Russian Far East, and of Abies gracilis, which is endemic to the Kamchatka Peninsula. Methods Nineteen populations were sampled for allozyme analysis. Seventeen of these populations were also screened for variation at two paternally inherited chloroplast DNA microsatellite loci (cpSSR) and variation at one maternally inherited mitochondrial marker (nad4‐3/4). Finally a subset of 11 populations was analysed with amplified fragment length polymorphism (AFLP). Comparisons were made with already available Abies sibirica data. For all sets of markers, we estimated genetic diversity and differentiation using an analysis of molecular variance (AMOVA). Population clustering was assessed with a Bayesian approach implemented in structure v.2.3. Results Among the three major species, A. sibirica, A. nephrolepis and A. sachalinensis, A. sachalinensis demonstrated the highest cytoplasmic and nuclear diversity and the most continental species, A. sibirica, the lowest. Both nuclear and mitochondrial DNA markers revealed the presence of a transitional zone on Sakhalin Island between A. nephrolepis and A. sachalinensis of south Sakhalin. The structure analysis delivered very clear results confirming the admixed origin of A. sachalinensis, with a genetic contribution from A. nephrolepis. No variation in cytoplasmic markers was found in A. gracilis, suggesting the occurrence of a recent bottleneck. Main conclusions There is a clear reduction of genetic diversity in Abies species from the Pacific coast into the continent. The higher diversity in A. sachalinensis could have two causes: a larger effective population size in the islands due to relatively stable climatic conditions and consequently less pronounced demographic fluctuations in population size and/or hybridization with continental and Japanese populations. Sakhalin Island is a major transitional zone for conifer species. Finally, the fir from Kamchatka, A. gracilis, should be regarded as a separate species closely related to the A. nephrolepis–A. sachalinensis complex.     

Gender identification of shovelnose sturgeon using ultrasonic and endoscopic imagery and the application of the method to the pallid sturgeon

Monthly sampling of shovelnose sturgeon Scaphirhynchus platorynchus , a biological surrogate for the endangered pallid sturgeon Scaphirhynchus albus , was conducted to develop a multi‐seasonal profile of reproductive stages. Data collected included histological characteristics of gonads from wild caught fish and laboratory and field ultrasonic and endoscopic images. These data were used to compare effectiveness of ultrasonic and endoscopic techniques at identifying gender of adult shovelnose sturgeon at different reproductive stages. The least invasive method ( i.e . ultrasound) was least effective while the most invasive ( i.e . endoscope through an abdominal incision) was the most effective at identifying shovelnose sturgeon gender. In most cases, success rate for identifying males was greater than females, with success at identifying both genders greater in more advanced reproductive stages. Concomitantly, for most months average reproductive stage was more advanced for males than females. April and May were the months with the most advanced reproductive stage, and were the months when ultrasound was most effective. Methods were also applied in the Upper Missouri River to validate their use on pallid sturgeon Scaphirhynchus albus . Ultrasound was successful at identifying pallid sturgeon gender, however, endoscopic examination through the urogenital duct was only successful at identifying pallid sturgeon gender when the urogenital duct was not opaque.     

Testicular development and serum sex steroid profiles during the annual sexual cycle of the male sturgeon hybrid the bester

Testicular development in the adult male F₁ sturgeon hybrid, the bester ( Huso huso L. female x Acipenser ruthenus L. male), was examined monthly in relation to serum sex steroid levels. Spermatogenesis lasted for 1 year, with meiosis generally starting in September and spermiogenesis in November, although there was considerable variation in testicular developmental stages between fish sampled monthly. Testicular development continued, slowly, during the winter months until April. Fish did not exhibit spontaneous spermiation, and phagocytotic activity of Sertoli cells became prominent from May onwards. Androgen levels increased during Spermatogenesis and remained high throughout the pre-spermiation period. In the degeneration stage, 11-ketotestosterone concentrations declined to low levels, while testoster- one levels remained high. The serum concentration of 17,20β-dihydroxy-4-pregnen-3-one was low throughout the reproductive cycle. Based on these results, it is suggested that the time appropriate for induction of final maturation would be from November–December to April when the testes are in the late stage of development.     

Rhyncholites and conchorhynchs as calcified jaw elements in some late Cretaceous ammonites

Conspicuous calcareous coverings are present in the anterior region of 17 fossil jaws from late Cretaceous rocks of Hokkaido (Japan) and Sakhalin (U.S.S.R.). The jaws were preserved in calcareous nodules either in situ in body chambers of ammonites or in close association with identifiable ammonite conch remains. From the morphologic similarity between in situ and isolated jaws, they may be attributed to Tetragonites glabrus, Gaudryceras tenuiliratum, G. denseplicatum, G. sp., and Neophylloceras subramosum. The jaw apparatus of these species is composed of two three-dimensional black walls of carbonate apatite, which might be a diagenetic replacement of chitinous material. The calcareous coverings in both upper and lower jaws closely resemble those of upper (rhyncholite) and lower (conchorhynch) jaws of modern Nautilus as well as rhyncholite and conchorhynch fossils in their gross morphology, microstructure, and chemical composition. Calcified remains of cephalopod jaws known as rhyncholites and conchorhynchs have been reported from late Paleozoic to Recent. The present discovery of ammonoid rhyncholites and conchorhynchs suggests that at least some previously known late Paleozoic and Mesozoic counterparts belong to the Ammonoidea. The essential similarity of jaw elements of some Late Cretaceous ammonites and modern Nautilus gives reliable information on the feeding habits of the former. The sharp and thick ammonoid rhyncholites and conchorhynchs may have had a special function for cutting up food, similar to those of Nautilus.     

Movement of sturgeons (Acipenser oxyrinchus oxyrinchus and A. fulvescens) in the St. Lawrence Estuary: A 35-year tagging study

Movement of Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) and lake sturgeon (A. fulvescens) in the St. Lawrence Estuary (Québec, Canada) are not fully understood. To assess the movement extent of both species, a mark–recapture study was conducted in collaboration with commercial fishermen operating in the St. Lawrence Estuary. Between 1981 and 2015, 3,367 Atlantic sturgeon (fork length 21.8–199.5 cm) and 3,180 lake sturgeon (fork length 17.8–190.8 cm) were tagged and released. Of these, 673 Atlantic sturgeon and 42 lake sturgeon were recaptured. The maximum distances traveled between capture and recapture locations were 1,307 km for Atlantic sturgeon (8 years after initial capture) and 252 km for lake sturgeon (less than 1 year after initial capture). Statistical analyses identified differences in the dispersal distance of both species as revealed by a first component characterized by individuals with short dispersal distances (98% and <35 km for Atlantic sturgeon; 58% and <1 km for lake sturgeon) and a second component characterized by individuals with longer dispersal distances (2% and >600 km for Atlantic sturgeon; 42% and >190 km for lake sturgeon). We suggest that the short dispersal distances detected in the vast majority of Atlantic sturgeon recaptures likely reflect strong site fidelity, highlighting the importance of the St. Lawrence Estuary as a preferred habitat for juveniles and subadults. Although recaptures were low for lake sturgeon because this species is only marginally targeted by commercial fishermen in the St. Lawrence Estuary, our results also showed that this species uses estuarine habitats and that half of the population seems to exhibit strong site fidelity (67% of individuals were recaptured within 2 km).     

Sex identification of morphologically-undifferentiated Siberian sturgeon with statistical analysis of gene expression patterns

The purpose of this study was to detail a simple strategy for sexing morphologically-undifferentiated fish using statistical analysis of gene expression patterns characterized by quantitative PCR. This approach is especially relevant for species without known genomic sex markers. The method was developed for early identification of female Siberian sturgeon as part of a genomics study. That study documented activation of the enzyme 17ß-hydroxy-steroid-dehydrogenase (hsd17b1) in future ovarian tissue at 3 months of age, concurrent with a small forkhead box L2 (foxl2) peak and emerging cytochrome P450, family 19, subfamily A (cyp19a1) expression. Major cyp19a1 and foxl2 peaks occurred in presumptive female gonads at 5–6 months. This pattern suggested a genetic relay mediating estrogen production throughout differentiation, possibly to maintain gonadal femininity. Genes involved in stem cell proliferation (lim homeobox 2 (lhx2)) and somatic-germ cell interaction maintenance (iroquois homeobox 5 (irx5) and iroquois homeobox 3 (irx3)) were also expressed during molecular differentiation, at 5–6 months. The roles of lhx2, irx3, and irx5 in fish sex differentiation should be confirmed using other methodologies. These results indicate that estrogens are crucial for ovarian differentiation in basal non-teleost fish, consistent with well-established patterns in teleosts, with hsd17b1 as one of the earliest biomarkers of gonadal development.     

Effects of stocking density during live transportation on haematological parameters of Siberian sturgeon (Acipenser baerii,Brandt, 1869)

In this study, we investigated the effects of different stocking densities water quality and blood parameters during the transportation of the Siberian sturgeon (Acipenser baerii). Experiments were carried out in high-density polyethylene tank with three different stocking densities (50, 100 and 150 kg/m³) for 20 h. Ammonium nitrogen (NH₄-N) and nitrite nitrogen (NO₂-N) were measured in the water samples. The erythrocyte, leucocyte, haematocrit (HCT), haemoglobin (Hb), cortisol, and sodium ion (Na⁺) were measured in blood samples. Ammonium nitrogen and nitrite nitrogen concentrations in the water have increased in parallel with the stocking density depending on the time. In the highest stocking density, the maximum levels of NH₄-N and NO₂-N at the 16^th h reached 2.64 mg/L and 4.74 mg/L, respectively. Erythrocyte, leucocyte, HCT, and Hb values did not differ significantly between the experimental groups (p > .05). The results showed that fish could be transported safely for 20 h in 15℃ water temperature at 50 kg/m³ stocking density; however, stocking density of 100 kg/m³ and over could threaten fish welfare and health as from 16^th h.     

Review of the Palaearctic Acomopterella Zaitzev (Diptera,Sciaroidea, Mycetophilidae)

The distribution of Acomopterella species in the Palaearctic region has been re-examined in this study, using recently collected material. The European species was found to be distributed in the eastern Palaearctic as well. A second Palaearctic species from Honshu (Japan) is herein described. The morphology of adult specimens was studied by light microscopy and scanning electron microscopy. The shape of functional specialized setae on mid tibiae in Acomopterella and seven further fungus gnat genera is described and the suitability of this character for systematic studies is discussed. Details of a “hind tibial organ” are described.The position of Acomopterella in the tribe Gnoristini is briefly discussed. Acomopterella is found to be more closely related to Speolepta Edwards, 1925, than to any other recent genus.     

Genetic monitoring of progeny derived from natural spawning of Acipenser ruthenus Linnaeus, 1758 and Acipenser stellatus Pallas, 1771 as well as their hybrids in the Lower Volga river during the period 2017 to 2019

The state of natural spawning of sturgeons in the Lower Volga River was investigated through genetic monitoring of juveniles (larvae, fingerlings, subadults) captured at seven traditionally used monitoring locations. Sampling was performed during the period 2017–2019. In total, 460 individuals were caught and genotyped using a set of six microsatellite loci (Afug41, Afug51, Afug135, An20, AoxD161, AoxD165) and mtDNA control region sequencing. The observed species-specific microsatellite alleles revealed the presence of juvenile sturgeons originating from natural spawning also included some hybrids between Acipenser stellatus and A. ruthenus. Thus, the majority of sterlet breeding takes place in natural spawning grounds but also sterlet hybridizes with a small number of stellate sturgeons. Juveniles of other sturgeon species were not identified, and it is concluded that natural spawning activities of Russian sturgeon, beluga and ship sturgeon were insignificant or even absent in the Lower Volga during the observation period.     

Improved genetic identification of acipenseriform embryos with application to the endangered pallid sturgeon Scaphirhynchus albus

We produced pallid sturgeon Scaphirhynchus albus embryos at five pre-hatch developmental stages and isolated and quantified genomic DNA from four of the stages using four commercial DNA isolation kits. Genomic DNA prepared using the kit that produced the largest yields and concentrations were used for microsatellite DNA analyses of 10–20 embryos at each of the five developmental stages. We attempted to genotype the hatchery-produced embryos at 19 microsatellite loci and confirmed reliable genotyping by comparing the microsatellite genotypes to those of known parents. Embryos at stages 5 and 8 did not produce reliable genotyping while those at stages 14, 24 and 33 did. We used the same DNA isolation method on 262 wild-caught acipenseriform embryos collected from the lower Yellowstone River. A total of 200 of the wild embryos were successfully identified to stages 8 to 34 and the rest could not be staged. Using a combination of single nucleotide polymorphism and microsatellite markers, 249 of the wild-caught embryos were genetically identified as paddlefish Polyodon spathula, five were identified as shovelnose sturgeon Scaphirhynchus platorynchus and eight failed to amplify. None were identified as pallid sturgeon. This study demonstrates that early-stage wild-spawned acipenseriform embryos can be genetically identified less than 24 h post-spawn. This methodology will be useful for recovery efforts for endangered pallid sturgeon and can be applied to other acipenseriform species.