The effect of diet on the structure of gut bacterial community of sympatric pair of whitefishes (Coregonus lavaretus): one story more

In the Coregonus lavaretus complex may be found lacustrine sympatric pairs, which serves as an intriguing model for studying different aspects of fish evolutionary biology. One such sympatric whitefish pair inhabits Teletskoye Lake (West Siberia, Russia) and includes a “large” form (Coregonus lavaretus pidschian (Gmelin, 1789)) and a “small” form (C. l. pravdinellus (Dulkeit, 1949)). C. l. pravdinellus has a narrow trophic specialization and feeds on zooplankton, whereas the diet of C. l. pidschian is based on benthic prey. In the present study we aimed to address the question of how the gut microbial community reflects the divergence in diet of a sympatric pair of whitefish. Studied samples included the mucosa and content were collected for cardiac and pyloric stomach, anterior, middle, and posterior intestine, but only mucosa was collected for the pyloric caeca. In addition, water, sediment, macrophyte (environmental microbiota) and invertebrate (microbiota of prey) samples were collected in the same location. The V3–V4 region of the 16S rRNA genes was chosen for microbiome analysis and the software PICRUSt used to estimate the difference functional roles of the microbiota. The number of OTUs and Chao1 index in mucosa and content of cardiac and pyloric stomach were significantly different between whitefish. Significant differences were observed between whitefish for content from different parts of the intestine in terms of OTU number and Chao1 indices, whereas for mucosa from the same parts of intestine these differences were absent. No significant differences were found for diversity estimates of mucosa and content of different parts of the gut (there were a few exceptions) between whitefish. The form of whitefish and the segment of the digestive system were factors with a significant determinative effect on the structure of the microbiota from gut mucosa and content. The most dominant phyla in mucosa and content of cardiac and pyloric stomach was Proteobacteria (57.0–84.0%) for both whitefish. Throughout the intestine of C. l. pidschian the dominant phyla in mucosa were Proteobacteria (38.8%) and Firmicutes (15.6%), whereas for C. l. pravdinellus–Tenericutes (49.6%) and Proteobacteria (28.1%). For both forms, the phylum Spirochaetes was found in a significant amount (20.0–25.0%) in the mucosa of the posterior intestine. While for the content obtained from anterior, middle and posterior intestines, the dominant bacterial phyla were the same as those described for mucosa from the same parts of the intestine for both whitefish. The bacterial community of the prey and environment was significantly different from bacterial communities found for all parts of the gut mucosa for both whitefish, with the exception of the mucosa of the cardiac stomach. According to PICRUSt the highest level of differences between whitefish at the L3 level were found for the intestinal mucosa (75.3%), whereas the lowest one was registered for stomach content (38.8%).info:eu-repo/semantics/publishedVersio

Dependence of element composition of bile of freshwater and marine fishes on some abiotic and biotic factors

Introduction: The elemental composition and physical properties of fish bile is a very important marker for ecotoxicological and physiological studies as bile is able to accumulate elements from environmental compartments. In the present study we focused on the effects of long term (seasons and years) and spatial distance (different water bodies) as well as other biotic (feeding habits, fish species, gut morphology) and abiotic (water salinity) factors on element composition of bile of 429 individuals belonging to 22 forms/species of fishes. Methods: Element composition was determined using an inductively coupled plasma optical emission spectrometry. The fishes were analyzed from one marine (the Kandalaksha Gulf of the White Sea), two freshwater oligotrophic (Teletskoye and Baunt lakes), and one freshwater eutrophic water body (Chany Lake). In parallel, we have estimated the volume and density of bile from the same fishes using an automatic pipette and electronic balance. Results and discussion: Based on fish bile density and volume data, we hypothesize that gastric fish species (those fishes that have a stomach) require a low volume of bile, but with higher density if compared to agastric fish species (those lacking a stomach) that mainly possessed large volumes of bile, but with a lower density. We have found that the concentrations of the following Na, K, Ca, Mg, S, P (major elements) and Al, Cu, Fe, Sr, Zn (trace elements) were the highest among the 28 elements studied. There were significant findings to contribute to a better understanding of the physiology of bile. First, we have determined the conditions formed in a water body in a given year (season) are more important for element composition of fish bile rather than other tested factors (feeding habits, gut morphology, etc.). Second, the feeding habits of fishes (carnivorous compared to non-carnivorous) had a significant effect based on differences among several elements. Third, the clear distinction between marine and freshwater fishes was found only for K and Na. Understanding which elements are produced as part of waste elimination and those that are present as a consequence of vital biological functions could improve the ecotoxicological study of bile as a marker of heavy metal contamination.The sample collection, expedition, and chemical analyses (for all fish except pike, perch, roach, and ide) were paid by the Russian Science Foundation (grant number 19-74-10054). The sample chemical analyses for pike and perch was supported by the Russian Foundation for Basic Research (grant number 19-34-60028) whereas for roach and ide was paid by Russian international scientific collaboration program Mega-grant (mega-grant № 075-15-2022-1134).info:eu-repo/semantics/publishedVersio

Trophic diversification and parasitic invasion as ecological niche modulators for gut microbiota of whitefish

Introduction: The impact of parasites on gut microbiota of the host is well documented, but the role of the relationship between the parasite and the host in the formation of the microbiota is poorly understood. This study has focused on the influence that trophic behavior and resulting parasitism has on the structure of the microbiome. Methods: Using 16S amplicon sequencing and newly developed methodological approaches, we characterize the gut microbiota of the sympatric pair of whitefish Coregonus lavaretus complex and the associated microbiota of cestodes parasitizing their intestine. The essence of the proposed approaches is, firstly, to use the method of successive washes of the microbiota from the cestode’s surfaces to analyze the degree of bacterial association to the tegument of the parasite. Secondly, to use a method combining the sampling of intestinal content and mucosa with the washout procedure from the mucosa to understand the real structure of the fish gut microbiota. Results and discussion: Our results demonstrate that additional microbial community in the intestine are formed by the parasitic helminths that caused the restructuring of the microbiota in infected fish compared to those uninfected. Using the desorption method in Ringer’s solution, we have demonstrated that Proteocephalus sp. cestodes possess their own microbial community which is put together from “surface” bacteria, and bacteria which are weakly and strongly associated with the tegument, bacteria obtained after treatment of the tegument with detergent, and bacteria obtained after removal of the tegument from the cestodes.info:eu-repo/semantics/publishedVersio

Trophic diversification and parasitic invasion as ecological niche modulators for gut microbiota of whitefish

Introduction: The impact of parasites on gut microbiota of the host is well documented, but the role of the relationship between the parasite and the host in the formation of the microbiota is poorly understood. This study has focused on the influence that trophic behavior and resulting parasitism has on the structure of the microbiome. Methods: Using 16S amplicon sequencing and newly developed methodological approaches, we characterize the gut microbiota of the sympatric pair of whitefish Coregonus lavaretus complex and the associated microbiota of cestodes parasitizing their intestine. The essence of the proposed approaches is, firstly, to use the method of successive washes of the microbiota from the cestode’s surfaces to analyze the degree of bacterial association to the tegument of the parasite. Secondly, to use a method combining the sampling of intestinal content and mucosa with the washout procedure from the mucosa to understand the real structure of the fish gut microbiota. Results and discussion: Our results demonstrate that additional microbial community in the intestine are formed by the parasitic helminths that caused the restructuring of the microbiota in infected fish compared to those uninfected. Using the desorption method in Ringer’s solution, we have demonstrated that Proteocephalus sp. cestodes possess their own microbial community which is put together from “surface” bacteria, and bacteria which are weakly and strongly associated with the tegument, bacteria obtained after treatment of the tegument with detergent, and bacteria obtained after removal of the tegument from the cestodes

Metapopulation Structure of Two Species of Pikeworm (Triaenophorus, Cestoda) Parasitizing the Postglacial Fish Community in an Oligotrophic Lake

In the present study, we estimated the levels of infestation of the main fish species that are hosts for two Triaenophorus species: T. crassus and T. nodulosus. The prevalence of T. crassus and T. nodulosus infestations in the intestine of their definitive host–pike Esox lucius was similar (71.0% and 77.4%, respectively). At the same time, the prevalence of T. crassus infestation in muscle tissue was significantly different between the second intermediate hosts, Coregonus lavaretus pidschian (31.4%) and Cor. l. pravdinellus (91.2%), due to considerable differences in their diets. For T. nodulosus, we found significant variations in the levels of prevalence among the second intermediate hosts—100% for Lota lota, 81.8% for Cottus sibiricus 31.9% for Thymallus arcticus, and 24.5% for Perca fluviatilis—that we also explained using different diets. Moreover, analysis of the symmetry of parasite infestations did not reveal any asymmetry between the number of cysts in the left and right body surfaces of the “planktivorous” form/species of whitefish, whereas in the ‘‘benthivorous”, an asymmetry of parasite infestations was found.The research of 2020 was partially supported by the Russian Foundation for Basic Research (grant number 19-34-60028), the data curation in 2021 was partially supported by the Russian Science Foundation (project No. 19-74-10054), and for 2022 was partially supported by the Russian international scientific collaboration program Mega-grant (mega-grant No. 075-15-2022-1134).info:eu-repo/semantics/publishedVersio

Chestnut shell tannins: effects on intestinal inflammation and dysbiosis in zebrafish

The aim of the present study was to test the possible ameliorative efficacy of phytochemicals such as tannins on intestinal inflammation and dysbiosis. The effect of a chestnut shell (Castanea sativa) extract (CSE) rich in polyphenols, mainly represented by tannins, on k-carrageenan-induced intestinal inflammation in adult zebrafish (Danio rerio) was tested in a feeding trial. Intestinal inflammation was induced by 0.1% k-carrageenan added to the diet for 10 days. CSE was administered for10 days after k-carrageenan induced inflammation. The intestinal morphology and histopathology, cytokine expression, and microbiota were analyzed. The k-carrageenan treatment led to gut lumen expansion, reduction of intestinal folds, and increase of the goblet cells number, accompanied by the upregulation of pro-inflammatory factors (TNFα, COX2) and alteration in the number and ratio of taxonomic groups of bacteria. CSE counteracted the inflammatory status enhancing the growth of health helpful bacteria (Enterobacteriaceae and Pseudomonas), decreasing the pro-inflammatory factors, and activating the anti-inflammatory cytokine IL-10. In conclusion, CSE acted as a prebiotic on zebrafish gut microbiota, sustaining the use of tannins as food additives to ameliorate the intestinal inflammation. Our results may be relevant for both aquaculture and medical clinic field

Dependence of pH values in the digestive tract of freshwater fishes on some abiotic and biotic factors

The values of pH in the digestive tracts of 20 freshwater fish species inhabiting various Russian Federation waterbodies were studied. Only in six species (Coregonus lavaretus, Coregonus migratorius, Catostomus catostomus, Carassius gibelio, Rutilus rutilus, Leuciscus leuciscus) out of 20 species, the differences in pH values between different regions of the intestine were significant. Feeding habits, feeding frequency, and gut fullness in fish affected pH values. Temperature was one of the most important factors affecting pH values. During cold seasons (spring and fall; average water temperature: 8–10 and 5–6○C, respectively), the pH values in fish guts were higher than that in the summer seasons(water temperature 22–25°C) for C. gibelio, Perca fluviatilis, Cyprinus carpio, L. leuciscus, and R. rutilus from the Chany Lake. Similar results (lower pH values in intestine at higher water temperatures) were also obtained for C. gibelio in warmer years in comparison to colder years in the same waterbody and in L. leuciscus and P. fluviatilis in the different waterbodies with different water temperatures. It is hypothesized that dependence of pH in fish gut on temperature may serve as a regulatory mechanism for maintaining the activities of hydrolytic enzymes at the required level for their successful functioning

Dependence of pH values in the digestive tract of freshwater fishes on some abiotic and biotic factors

The values of pH in the digestive tracts of 20 freshwater fish species inhabiting various Russian Federation waterbodies were studied. Only in six species (Coregonus lavaretus, Coregonus migratorius, Catostomus catostomus, Carassius gibelio, Rutilus rutilus, Leuciscus leuciscus) out of 20 species, the differences in pH values between different regions of the intestine were significant. Feeding habits, feeding frequency, and gut fullness in fish affected pH values. Temperature was one of the most important factors affecting pH values. During cold seasons (spring and fall; average water temperature: 8–10 and 5–6○C, respectively), the pH values in fish guts were higher than that in the summer seasons(water temperature 22–25°C) for C. gibelio, Perca fluviatilis, Cyprinus carpio, L. leuciscus, and R. rutilus from the Chany Lake. Similar results (lower pH values in intestine at higher water temperatures) were also obtained for C. gibelio in warmer years in comparison to colder years in the same waterbody and in L. leuciscus and P. fluviatilis in the different waterbodies with different water temperatures. It is hypothesized that dependence of pH in fish gut on temperature may serve as a regulatory mechanism for maintaining the activities of hydrolytic enzymes at the required level for their successful functioning

DNA extraction protocols may influence biodiversity detected in the intestinal microbiome: a case study from wild Prussian carp, Carassius gibelio

In this investigation, we examined the influence of different DNA extraction protocols on results obtained for intestinal microbiota of Prussian carp. We showed that significant differences were observed in numbers of reads, OTUs, Shannon index and taxonomic composition between two different DNA extraction protocols for intestine of Prussian carp (Carassius gibelio), and differences were also evident between microbial communities in the intestinal mucosa and intestinal content. Statistical analyses of 25 published articles also revealed a significant relationship between methods of DNA extraction and bacterial diversity in fish intestine of freshwater species. Microbial diversity, community structure, proportions of read numbers derived from each OTU and the total number of OTU's obtained by different DNA extraction protocols could lead to a bias in results obtained in some cases, and therefore researchers should be conservative in conclusions about community structures

Seasonal changes in kinetic parameters of trypsin in gastric and agastric fish

The objective of the present study was to assess if trypsin, a key enzyme involved in protein digestion, presents some kind of functional adaptations to seasonal changes in water temperature in freshwater fish. In order to test this hypothesis, individuals of two fish species Carassius gibelio (agastric) and Perca fluviatilis (gastric) were sampled in the basin of Chany Lake (Siberia, Russia) at two different seasons (spring and summer). Apparent kinetic parameters (Km and Vmax) were determined for both species and seasons at the actual pH values in fish guts, and at actual temperatures. Results showed a significant effect of both the species and sampling season on the apparent kinetic parameters of trypsin. In the case of Prussian carp, Km and Vmax were lower for each assayed temperature (for 5 and 15 °C the differences were significant) for fish sampled in summer when compared to those sampled in spring. In contrast, values of Km in perch tended to be lower in spring at 5 and 25 °C but these differences were not significant, while Vmax showed a significant decrease in summer samples. This suggests a sort of functional adaptation of the same trypsin enzymes to seasonal changes, oriented to maximize protein digestion under variable conditions