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Simple Summary: Novel optical sensors require implantation into the most transparent organs in order to ensure the most reliable and rapid monitoring of animal health. Widely farmed salmonid fish, such as rainbow trout, have highly translucent adipose fin, which we tested here and showed its high potential as the implantation site for the fluorescent sensors. The filamentous sensors were convenient to inject into the fin, and their optical signal was easily detectable using a simple hand-held device even without immobilization of the fish. Responsiveness of the sensors inside the adipose fin to bodily changes was shown under induced acidosis of fish fluids. The obtained results characterize adipose fin as the favorable site for implantation of optical sensors into salmonids for real-time tracking animal physiological status in basic research and aquaculture. Implantable optical sensors are emerging tools that have the potential to enable constant real-time monitoring of various internal physiological parameters. Such a possibility will open new horizons for health control not only in medicine, but also in animal husbandry, including aquaculture. In this study, we analyze different organs of commonly farmed rainbow trout (Oncorhynchus mykiss) as implantation sites for fluorescent sensors and propose the adipose fin, lacking an endoskeleton, as the optimal choice. The fin is highly translucent due to significantly thinner dermis, which makes the detectable fluorescence of an implanted sensor operating at the visible light range by more than an order of magnitude higher relative to the skin. Compared to the proximal parts of ray fins, the adipose fin provides easy implantation and visualization of the sensor. Finally, we tested fluorescent pH sensors inside the adipose fin and demonstrated the possibility of acquiring their signal with a simple hand-held device and without fish anesthesia. All these features will most likely make the adipose fin the main "window" into the internal physiological processes of salmonid fish with the help of implantable optical sensors. [ABSTRACT FROM AUTHOR]

Implantable sensors based on shaped biocompatible hydrogels are now being extensively developed for various physiological tasks, but they are usually difficult to implant into small animals. In this study, we tested the long-term in vivo functionality of pH-sensitive implants based on amorphous 2.7% polyacrylamide hydrogel with the microencapsulated fluorescent probe SNARF-1. The sensor was easy to manufacture and introduce into the tissues of a small fish Danio rerio, which is the common model object in biomedical research. Histological examination revealed partial degradation of the gel by the 7th day after injection, but it was not the case on the 1st day. Using the hydrogel sensor, we were able to trace the interstitial pH in the fish muscles under normal and hypercapnic conditions for at least two days after the implantation. Thus, despite later immune response, amorphous polyacrylamide is fully suitable for preparing implantable sensors for various mid-term physiological experiments on small fishes. The proposed approach can be further developed to create implantable sensors for animals with similar anatomy. [ABSTRACT FROM AUTHOR]

Ancient lakes are known speciation hotspots. One of the most speciose groups in the ancient Lake Baikal are gammaroid amphipods (Crustacea: Amphipoda: Gammaroidea). There are over 350 morphological species and subspecies of amphipods in Baikal, but the extent of cryptic variation is still unclear. One of the most common species in the littoral zone of the lake, Eulimnogammarus verrucosus (Gerstfeldt, 1858), was recently found to comprise at least three (pseudo)cryptic species based on molecular data. Here, we further explored these species by analyzing their mitogenome-based phylogeny, genome sizes with flow cytometry, and their reproductive compatibility. We found divergent times of millions of years and different genome sizes in the three species (6.1, 6.9 and 8 pg), further confirming their genetic separation. Experimental crossing of the western and southern species, which are morphologically indistinguishable and have adjacent ranges, showed their separation with a post-zygotic reproductive barrier, as hybrid embryos stopped developing roughly at the onset of gastrulation. Thus, the previously applied barcoding approach effectively indicated the separate biological species within E. verrucosus. These results provide new data for investigating genome evolution and highlight the need for precise tracking of the sample origin in any studies in this morphospecies. [ABSTRACT FROM AUTHOR]

Simple Summary: Deep-water habitats are the largest ecosystem on the planet: over half of the Earth's surface is covered with a water layer deeper than 200 m and remains poorly explored. Lake Baikal is the only freshwater body inhabited by animals adapted to the deep-water zone independently from their marine counterparts. Comparing these convergently evolved freshwater and marine animals is invaluable for revealing the basic mechanisms of adaptation to high hydrostatic pressure. However, laboratory experiments on deep-water organisms still usually require lifting them to the water's surface and exposing them to potentially hazardous decompression, while endemics from Lake Baikal are poorly studied in this regard. Here, we compared metabolic reactions to such pressure decreases in two Baikal deep-water amphipods (shrimp-like crustaceans) from the genus Ommatogammarus: one species is known to tolerate pressures close to atmospheric levels, while the second was only observed at the pressures from 5 atm and above. We expected that the energy metabolism of the shallower-dwelling species would function better under the atmospheric pressure but found no substantial differences. Thus, despite some difference in long-term survival at atmospheric pressure, both species are suitable for laboratory studies as freshwater model objects adapted to large pressure variations. Lake Baikal is the only freshwater reservoir inhabited by deep-water fauna, which originated mostly from shallow-water ancestors. Ommatogammarus flavus and O. albinus are endemic scavenger amphipods (Amphipoda, Crustacea) dwelling in wide depth ranges of the lake covering over 1300 m. O. flavus had been previously collected close to the surface, while O. albinus has never been found above the depth of 47 m. Since O. albinus is a promising model species for various research, here we tested whether O. albinus is less metabolically adapted to atmospheric pressure than O. flavus. We analyzed a number of energy-related traits (contents of glucose, glycogen and adenylates, as well as lactate dehydrogenase activity) and oxidative stress markers (activities of antioxidant enzymes and levels of lipid peroxidation products) after sampling from different depths and after both species' acclimation to atmospheric pressure. The analyses were repeated in two independent sampling campaigns. We found no consistent signs of metabolic disturbances or oxidative stress in both species right after lifting. Despite O. flavus surviving slightly better in laboratory conditions, during long-term acclimation, both species showed comparable reactions without critical changes. Thus, the obtained data favor using O. albinus along with O. flavus for physiological research under laboratory conditions. [ABSTRACT FROM AUTHOR]

Layer-by-layer assembled microcapsules are promising carriers for the delivery of various pharmaceutical and sensing substances into specific organs of different animals, but their utility in vivo inside such an important group as crustaceans remains poorly explored. In the current study, we analyzed several significant aspects of the application of fluorescent microcapsules covered by polyethylene glycol (PEG) inside the crustacean circulatory system, using the example of the amphipod Eulimnogammarus verrucosus. In particular, we explored the distribution dynamics of visible microcapsules after injection into the main hemolymph vessel; analyzed the most significant features of E. verrucosus autofluorescence; monitored amphipod mortality and biochemical markers of stress response after microcapsule injection, as well as the healing of the injection wound; and finally, we studied the immune response to the microcapsules. The visibility of microcapsules decreased with time, however, the central hemolymph vessel was confirmed to be the most promising organ for detecting the spectral signal of implanted microencapsulated fluorescent probes. One million injected microcapsules (sufficient for detecting stable fluorescence during the first hours after injection) showed no toxicity for six weeks, but in vitro amphipod immune cells recognize the PEG-coated microcapsules as foreign bodies and try to isolate them by 12 h after contact. [ABSTRACT FROM AUTHOR]