Effects of different temperatures on Leiocassis longirostris gill structure and intestinal microbial composition.

Fish are poikilothermic vertebrates and their physiological activities are affected by water temperature. In recent years, extreme weather has occurred frequently, and temperature changes have adversely affected the growth of farmed fish. To explore the changes in gill tissue structure caused by changing the water temperature and the relationship between the intestinal microbiota and the Leiocassis longirostris host adaptation mechanism, gill tissue sections and intestinal microbial 16S rRNA amplicon sequencing were conducted under different temperature stress (low temperature 4 °C, normal temperature 26 °C and high temperature 32 °C). The results showed that heat stress and cold stress caused injury and swelling, terminal congestion, cell vacuolation, and necrosis of the gill tissue of L. longirostris. For intestinal microbiota, the abundances of Pseudomonadota and Bacillota increased at the cold stress, while the abundances of Fusobacteriota and Bacteroidota increased at the heat stress. The number of opportunistic bacteria, mainly Aeromonas and Acinetobacter, was the highest under cold stress. In addition, the richness of the intestinal microbiota decreased significantly at heat and cold stresses, while evenness increased. Prediction of intestinal microbiota function showed that most common functions, such as metabolism of cofactors and vitamins, energy metabolism and replication and repair, were decreased significantly at heat stress and cold stress, and phylogenetic relationship analysis revealed significant differences among the groups. In conclusion, the change of temperature altered the gill tissue structure, and affected the structure and homeostasis of the intestinal microbiota, thus affecting the survival time of L. longirostris, and cold stress had a greater effect than heat stress.

Enzymatic Stress Responses of Coreius guichenoti to Microplastics with Different Particle Sizes.

The wild population resources of Coreius guichenoti have sharply declined in recent decades, and any negative factors may have a significant impact on their survival. In this study, the enzymatic stress responses of C. guichenoti to 25 and 48 µm polyethylene fragments were explored for the first time. This was achieved by evaluating the changes in physiological and biochemical indicators of the species in response to the environmental stimuli of microplastics. In this study, we observed an early stress response in the external tissues of C. guichenoti following exposure to microplastics. The TP content in skin and muscle and the MDA content in skin, gill and muscle initially showed a significant increase. The skin, gill, and muscle exhibited greater stress responses to M5 particles, whereas M3 particles caused a greater response in the intestine and especially the liver. After the removal of microplastic exposure, the stress state of the C. guichenoti would be alleviated in a short period, but it could not fully recover to the pre-exposure level. In summary, microplastics pose a significant threat to C. guichenoti. While their negative effects can be alleviated by the removal of microplastics exposure, full recovery does not occur in a short period. Continuous monitoring of microplastics in natural waters and targeted aquatic ecological restoration are essential to ensure the normal growth and reproduction of the wild population of C. guichenoti.

Single-nuclei RNA-seq reveals skin cell responses to Aeromonas hydrophila infection in Chinese longsnout catfish Leiocassis longirostris.

As the primary natural barrier that protects against adverse environmental conditions, the skin plays a crucial role in the innate immune response of fish, particularly in relation to bacterial infections. However, due to the diverse functionality and intricate anatomical and cellular composition of the skin, deciphering the immune response of the host is a challenging task. In this study, single nuclei RNA-sequencing (snRNA-seq) was performed on skin biopsies obtained from Chinese longsnout catfish (Leiocassis longirostris), comparing Aeromonas hydrophila-infected subjects to healthy control subjects. A total of 19,581 single nuclei cells were sequenced using 10x Genomics (10,400 in the control group and 9,181 in the treated group). Based on expressed unique transcriptional profiles, 33 cell clusters were identified and classified into 12 cell types including keratinocyte (KC), fibroblast (FB), endothelial cells (EC), secretory cells (SC), immune cells, smooth muscle cells (SMC), and other cells such as pericyte (PC), brush cell (BC), red blood cell (RBC), neuroendocrine cell (NDC), neuron cells (NC), and melanocyte (MC). Among these, three clusters of KCs, namely, KC1, KC2, and KC5 exhibited significant expansion after A. hydrophila infection. Analysis of pathway enrichment revealed that KC1 was primarily involved in environmental signal transduction, KC2 was primarily involved in endocrine function, and KC5 was primarily involved in metabolism. Finally, our findings suggest that neutrophils may play a crucial role in combating A. hydrophila infections. In summary, this study not only provides the first detailed comprehensive map of all cell types present in the skin of teleost fish but also sheds light on the immune response mechanism of the skin following A. hydrophila infection in Chinese longsnout catfish.

Study on the Adaptive Regulation of Light on the Stress Response of Mandarin Fish (Siniperca chuatsi) with Re-Feeding after Starvation.

Light influences the stress response to environmental stimuli and feeding behaviors of Siniperca chuatsi and, thus, is an important regulator of normal growth and development. In this study, we first explored the important role of light on the digestive and stress capacity of S. chuatsi by studying the changes in physiological and biochemical indicators of S. chuatsi, taking the re-feeding after starvation as the constant environmental stimulus and the light intensity as the adjustable environmental stimulus. The activity of protease and lipase was generally higher in the stomach tissues than in the intestinal tissues, especially lipase, which was higher in stomach tissues under all light conditions, and the protease and lipase activity peaked in the stomach tissues of S. chuatsi at a light intensity of 18.44 ± 3.00 lx and in intestinal tissues at 11.15 ± 2.01 lx, respectively, indicating that greater light intensity increased the digestive capacity of stomach tissues, whereas lower light intensity facilitated the digestive capacity of intestinal tissues. The tissues of the gill, stomach, and intestine had relatively high activity of stress-related enzymes, whereas the tissues of the brain, kidney, liver, and plasma samples had relatively low activity of enzymes. Collectively, the results show that light intensity at 11.15 ± 2.01 lx promoted digestive capacity in the intestine and enhanced the anti-stress ability of S. chuatsi in response to stress induced by re-feeding after starvation. These findings should prove useful for artificial breeding of S. chuatsi.

PacBio single-molecule long-read sequencing provides new insights into the complexity of full-length transcripts in oriental river prawn, macrobrachium nipponense.

BACKGROUND: Oriental river prawn (Macrobrachium nipponense) is one of the most dominant species in shrimp farming in China, which is a rich source of protein and contributes to a significant impact on the quality of human life. Thus, more complete and accurate annotation of gene models are important for the breeding research of oriental river prawn. RESULTS: A full-length transcriptome of oriental river prawn muscle was obtained using the PacBio Sequel platform. Then, 37.99 Gb of subreads were sequenced, including 584,498 circular consensus sequences, among which 512,216 were full length non-chimeric sequences. After Illumina-based correction of long PacBio reads, 6,599 error-corrected isoforms were identified. Transcriptome structural analysis revealed 2,263 and 2,555 alternative splicing (AS) events and alternative polyadenylation (APA) sites, respectively. In total, 620 novel genes (NGs), 197 putative transcription factors (TFs), and 291 novel long non-coding RNAs (lncRNAs) were identified. CONCLUSIONS: In summary, this study offers novel insights into the transcriptome complexity and diversity of this prawn species, and provides valuable information for understanding the genomic structure and improving the draft genome annotation of oriental river prawn.

Integrated analysis of how gender and body weight affect the intestinal microbial diversity of Gymnocypris chilianensis.

Intestinal microorganisms that living in the mucosa and contents of the gastrointestinal tract of animals, have close links with their hosts over a long evolutionary history. The community structure of the fish intestinal microbiota is associated with food, living environment, and the growth stage. To screen for potential probiotics that can be used for regulating breeding behaviors, this study focused on the diversity of fish intestinal microorganisms. This study aimed to investigate the effects of sex and body weight on the intestinal microbial diversity of Gymnocypris chilianensis in the wild. The results showed that the significant high diversity and richness of intestinal microbiota were fould in heavier individuals, and males. The dominant bacterial phyla of G. chilianensis were Proteobacteria, Firmicutes, and Bacteroidetes. In addition, the abundance of Firmicutes varied significantly among different body weights. The genus profile revealed that small individuals were dominated by Weissella, while females were dominated by Aeromonas, and both large individuals and males were dominated by other genera. Phylogenetic relationships and UPGMA clustering analysis showed significant differences among the groups. In general, the two main factors that have an effect on the intestinal microbiota diversity of wild G. chilianensis are sex and body weight.

Studies on the molecular level changes and potential resistance mechanism of Coreius guichenoti under temperature stimulation.

In this study, we used transcriptome and proteome technology to analyze molecular level changes in tissues of Coreius guichenoti cultured at high temperature (HT) and low temperature (LT). We also screened for specific anti-stress genes and proteins and evaluated the relationships between them. We identified 201,803 unigenes and 10,623 proteins. Compared with the normal temperature (NT), 408 genes and 1,204 proteins were up- or down-regulated in brain tissues, respectively, at HT, and the numbers were 8 and 149 at LT. In gill tissues, the numbers were 101 and 1,745 at HT and 27 and 511 at LT. In gill tissues at both temperatures, the degree of down-regulation (average, HT 204.67-fold, LT 443.13-fold) was much greater than that of up-regulation (average, HT 28.69-fold, LT 17.68-fold). The protein expression in brain (average, up 52.67-fold, down 13.54-fold) and gill (average, up 73.02-fold, down 12.92-fold) tissues increased more at HT than at LT. The protein expression in brain (up 3.77-fold, down 4.79-fold) tissues decreased more at LT than at HT, whereas the protein expression in gill (up 8.64-fold, down 4.35-fold) tissues was up-regulated more at LT than at HT. At HT, brain tissues were mainly enriched in pathways related to metabolism and DNA repair; at LT, they were mainly enriched in cancer-related pathways. At both temperatures, gill tissues were mainly enriched in pathways related to cell proliferation, apoptosis, immunity, and inflammation. Additionally, Kyoto Encyclopedia of Genes and Genomes pathway analysis showed more differentially expressed proteins in gill tissues than in brain tissues at HT and LT, and temperature stimulation led to the strengthening of metabolic pathways in both tissues. Of the 96 genes we identified as potentially being highly related to temperature stress (59 from transcriptome and 38 from proteome data), we detected heat shock protein 70 in both the transcriptome and proteome. Our results improved our understanding of the differential relationship between gene expression and protein expression in C. guichenoti. Identifying important temperature stress genes will help lay a foundation for cultivating C. guichenoti, and even other fish species, that are resistant to HT or LT.

Toxic metals in rice-fish co-culture systems and human health.

The threat of contamination with toxic metals (TMs) to food security and human health has become a high priority in recent decades. Hence, countless studies have investigated the safe cultivation of rice and fish, respectively, as the main food crop and protein source worldwide. For the present study, a literature search of the PubMed, Web of Science, ScienceDirect, and China National Knowledge Infrastructure databases identified 11 studies that met the inclusion criteria and provided sufficient data to assess the relationship between TM contamination of rice, fish, and shrimp products from rice-fish co-culture systems and carcinogenic risk (CR) and non-carcinogenic risk (non-CR) to humans. The result showed that consumption of Monopterus albus and rice contaminated with a single TM had a slight non-CR, which is synergistically increased by multiple TMs. Consumption of Procambarus clarkii posed no non-CR to humans. The CR of all studies ranged from 1.77 × 10-10 to 5.65 × 10-8, and less than 1 × 10-6, indicating that under current food safety guidelines, rice and fish produced by rice-fish co-culture systems pose no CR. Rice-fish co-culture systems can greatly reduce the CR and non-CR of TMs to humans.

Toxic Metals in a Paddy Field System: A Review.

The threat of toxic metals to food security and human health has become a high-priority issue in recent decades. As the world's main food crop source, the safe cultivation of rice has been the focus of much research, particularly the restoration of toxic metals in paddy fields. Therefore, in this paper, we focus on the effects of toxic metals on rice, as well as the removal or repair methods of toxic metals in paddy fields. We also provide a detailed discussion of the sources and monitoring methods of toxic metals pollution, the current toxic metal removal, and remediation methods in paddy fields. Finally, several important research issues related to toxic metals in paddy field systems are proposed for future work. The review has an important guiding role for the future of heavy metal remediation in paddy fields, safe production of rice, green ecological fish culture, and human food security and health.

Integrated analysis of the miRNA and mRNA expression profiles in Leiocassis longirostris at gonadal maturation.

Leiocassis longirostris is a commercially important fish species that shows a sexually dimorphic growth pattern. A lack of molecular data from the gonads of this species has hindered research and selective breeding efforts. In this study, we conducted a comprehensive analysis of the expression profile of miRNA and mRNA to explore their regulatory roles in the gonadal maturation stage of L. longirostris. We identified 60 differentially expressed miRNAs and 20,752 differentially expressed genes by sequencing. A total of 90 miRNAs and 21 target genes involved in gonad development and sex determination were identified. Overall, the results of this study enhance our understanding of the molecular mechanisms underlying sex determination and differentiation and provide valuable genomic information for the selective breeding of L. longirostris.

Genetic diversity of two color morphs of Northern snakehead (Channa argus) unveiled by the mitochondrial DNA D-loop region.

To analyze the genetic background of 'white' type Northern snakehead (Channa argus), and provide atheoretical basis for breeding of C. argus, the investigation of genetic diversity and population structure were investigated based on the complete sequences of mitochondrial DNA D-loop region for three cultured 'white' type C. argus populations, and four 'bicolor' type C. argus populations were used to compare with them; 28 mutation loci and 30 haplotypes were found in the D-loop sequence of all individuals with a total length of 907 bp. The highest haplotype diversity (Hd ) and nucleotide diversity (Pi ) in the 'white' type C. argus populations were 0.505 and 0.00057, respectively, which lower than those in the 'bicolor' type C. argus populations (Hd = 0.911, Pi = 0.00326). Population differentiation values (F ST) show that the four 'bicolor' type C. argus populations had obvious genetic differentiation (Fst: 0.21902-0.49428. p < 0.01), but not in the three 'white' type C. argus populations (Fst: -0.00571 to 0.07261. p > 0.05). The phylogenetic tree and Median Joining (MJ) network showed that the genetic distance among 'white' type C. argus populations is very close. Therefore, much attention should be paid to protecting population genetic diversity and avoiding inbreeding in the breeding of 'white' type C. argus.

Integrative analysis identifies the quality advantage and corresponding regulatory mechanism of paddy field-cultured crayfish (Procambarus clarkii).

Procambarus clarkii is the dominant economic variety of crayfish in China, and paddy field shrimp cultivation is an organic mode of traditional rice-fish cultivation, with paddy field shrimp being the country's prevailing aquatic product. However, little has been reported on the differences in meat quality and digestive ability between paddy field and pond fish. In this study, the muscle composition and digestive function regulation of P. clarkii in ponds and paddies were studied to explore the influence of paddy field culture on P. clarkii quality. The results showed that the muscle composition of paddy field shrimp was significantly changed, with increased protein and decreased lipid levels. Through the study of the hepatopancreas and intestinal microbial diversity of P. clarkii, we hypothesized that rice farming may cause changes in its bacterial spectrum, stimulate the digestive functions of its intestines and hepatopancreas, cause differential expression of multi-substance metabolic pathways, and ultimately result in the substances' deposition in its muscles. This study revealed the impact of rice cultivation on P. clarkii from the perspective of meta-metabolism, and it demonstrated the advantages of paddy field shrimp cultivation.Key points• We explored the influence of paddy field culture on P. clarkii quality.• Muscle composition of paddy field shrimp was significantly changed, with increased protein and decreased lipid levels in paddy field.• Rice farming caused changes in its bacterial spectrum and stimulated the digestive functions of hepatopancreas.

Growth and nutrient uptake of Myriophyllum spicatum under different nutrient conditions and its potential ecosystem services in an enclosed sea area in the East China Sea.

We investigated the growth and nutrient uptake of Myriophyllum spicatum under different nutrient conditions and evaluated its implications for ecosystem services in an enclosed area of Jinshan. The specific growth rate ranged from 1.29%-4.37%/day, and the dissolved inorganic carbon and nitrogen, and phosphorus uptake rates were 1.30-1.62, 0.040-0.453, and 0.003-0.027 mg/(g∙day), respectively, under different nutrient conditions. The O2-production and carbon-sequestration efficiencies in the field were 154.30 and 1.25 mg/(g DW∙h), respectively. The average removal efficiencies of NH4+-N, NO3--N, NO2--N, and PO43--P were 43.05%, 97.03%, 64.26%, and 59.24%, respectively, in M. spicatum-cultivated areas compared with in the open sea. Harvesting of M. spicatum removed 12,936.87, 1289.97 and 114.81 kg of carbon, nitrogen, and phosphorus, respectively, from seawater in Jinshan in Nov, 2018. In conclusion, M. spicatum is a good candidate for integrated macrophyte/animal multi-trophic aquaculture in terms of nutrient extraction and economic diversification in low-salinity environments.

Differences in the digestive enzyme activity, intestinal mucosa and microbial community in loach cultivated in two separate environments.

BACKGROUND: Fish culture in rice paddies can contribute to increasing yields of rice and surplus fish products. Environmental impacts and food-safety issues have become important topics in aquaculture, and organic foods currently were paid attention by researchers and industry practitioners. But the mechanism of differences in quality of Loach (Paramisgurnus dabryanus) reared in rice fields and ponds remains largely uncharacterized. In this study,digestive enzyme activity, intestinal mucosa cells and the gut microbial community of loach were determined under the two separate cultivation modes. RESULTS: The levels of intestinal digestive enzyme activity of fish reared in the paddy-cultivated mode (PACM) were higher (P < 0.05) than those in the pond-cultivated mode (POCM). It was extremely significant (P < 0.01) for the activity of lipase in the liver, foregut and midgut, and for the activities of amylase and trypsin in the hindgut. Acid mucous cells in the loach foregut in PACM were fewer than in POCM (P < 0.01). In summer, the abundance of the Firmicutes, Lactobacillus spp., Aeromonas hydrophila, Enterobacteriaceae and Streptococcus spp. in loach intestinal mucosa in PACM was higher than in POCM. In fall, the abundance of total bacteria, the Bacteroidetes, Bifidobacterium and Enterobacteriaceae in the intestinal mucosa in PACM was likewise higher than in POCM. These differences were significant (P < 0.05 or P < 0.01) between loach in the two separate culture modes for all microorganisms except for A. hydrophila and Streptococcus spp. In addition, quantitative PCR assays showed that some microorganisms presented consistently similar abundances in the gut as in the culture water. CONCLUSIONS: These results showed some enzymatic activities involved in digestion in liver and intestine of loach in PACM were higher than those in POCM, as using digestive enzyme analysis and histological observation of intestinal sections. These findings suggest most of the microorganisms examined in the gut mucosa of loach in the two culture modes significantly differed in abundance between summer and fall. However, some pathogenic bacteria in the gut, particularly A. hydrophila, presented lower abundance in PACM in fall, yet did not differ in abundance between loach in the two cultivation modes.

Observational comparisons of intestinal microbiota characterizations, immune enzyme activities, and muscle amino acid compositions of loach in paddy fields and ponds in Sichuan Province.

A balanced intestinal microbial ecosystem is crucial for the growth and health of animals because it can influence the digestion and absorption of nutrients in the intestine. Different culture conditions may change the ecology of microbial in intestine and thus affect the overall growth performance of an animal. In this study, we compared intestinal morphologies, microbiota characterizations, immune enzyme activities, and muscle amino acid compositions of loach cultured in paddy fields and ponds. The fish were fed with the same diets from May 5 to November 5 (2015) in three paddy or ponds. Fish samples were collected for analysis in the August (summer season) and November (fall season) during the feeding trial. In both culture conditions, results based on microscopy observation showed that the intestinal perimeter, fold height, fold radical, and total absorption of the gut were significantly higher in the foregut than that found in the midgut and hindgut (P < 0.01). The average final body weight of fish was similar between the two culture conditions (P > 0.05). The percentage of carcass weight to whole loach weight for samples collected from paddy field (91.6 ± 1.1) was significantly higher than the index measured for loach from pond (87.3 ± 3.4, P < 0.05). Results based on denaturing gradient gel electrophoresis demonstrated that the Shannon-diversity index, evenness, and richness of intestinal flora were increased from summer to fall in paddy cultivation. In pond culture condition, however, the above indexes obtained from mucosa and intestinal contents decreased in fish from summer to fall. The sequencing results of bands indicated that the predominant microorganisms are Proteobacteria, Firmicutes, and Actinobacteria in the intestine of fish being cultured in both cultures. Activities of alkaline phosphatase (AKP, in two culture conditions) and superoxide dismutase (SOD, in paddy field) presented a gradual decrease trend from foregut to hindgut of fish. The activities of acid phosphatase (ACP, in midgut), AKP (in midgut and hindgut), SOD (in foregut), and lysozyme (LZM, in midgut) were significantly higher in fish cultured in paddy than those in pond (P < 0.01). In addition, the percentage of some essential amino acids (valine, methionine, and phenylalanine) based on total amino acids in muscle was significantly higher in fish cultured in paddies than in ponds. In summary, the fish cultured in paddy or pond was not significantly different in growth but the two culture conditions seems to generate different carcass yield and changed the amino acid profiles of fish muscle. The similar predominance microorganisms were identified in the intestine of fish from two conditions, and the quantification of microbial in the intestine will be determined in the future, but part activities involved in immune protection were higher for fish cultured in paddy fields.

The effect of high pressure on the intracellular trehalose synthase activity of Thermus aquaticus.

To understand the effect of high pressure on the intracellular trehalose synthase activity, Thermus aquaticus (T. aquaticus) in the logarithmic growth phase was treated with high-pressure air, and its intracellular trehalose synthase (TSase) activity was determined. Our results indicated that pressure is a factor strongly affecting the cell growth. High pressure significantly attenuated the growth rate of T. aquaticus and shortened the duration of stationary phase. However, after 2 h of culture under 1.0 MPa pressure, the activity of intracellular TSase in T. aquaticus reached its maximum value, indicating that pressure can significantly increase the activity of intracellular TSase in T. aquaticus. Thus the present study provides an important guide for the enzymatic production of trehalose.