Introduction:Bamboo clum powder is an unconventional feed ingredient rich in insoluble dietary fiber, and its potential application and optimal inclusion level in laying hen diets remain unclear. This study was conducted to evaluate the effects of dietary supplementation with micronized bamboo powder (MBP) on laying performance, apparent nutrient digestibility, intestinal morphology, and cecal microbiota in late-laying hens, and to determine its appropriate inclusion level. Methods:A total of 480 42-week-old Hy-Line Brown laying hens were randomly allocated into 4 groups with 5 replicates of 24 hens each. The trial hens were fed a basal diet (CON) or basal diets supplemented with 2% (TRE1), 3% (TRE2), or 4% (TRE3) MBP for 28 days following a 7-day adaptation period. Results:Compared with the CON group, the TRE3 group exhibited a significantly lower laying rate during days 15-28 and the overall experimental period of 1-28 d (p < 0.05), while no significant differences were observed between the TRE1, TRE2, and CON groups (p > 0.05). Moreover, the TRE3 group had significantly lower egg mass during days 15-28 and days 1-28 compared with the CON and TRE1 groups (p < 0.05). Treatments had no significant effect on serum triglyceride, total cholesterol, total protein or glucose levels (p > 0.05). The TRE3 group showed significantly lower apparent digestibility of dry matter, crude ash, crude protein, and gross energy compared to the CON group (p < 0.05). Intestinal histological analysis revealed that the TRE3 group had significantly greater duodenal and ileal wall thickness than the CON group (p < 0.05). Conversely, the TRE1 and TRE2 groups exhibited significantly higher jejunal villus height compared to the CON group (p < 0.05). 16S rRNA sequencing analysis of cecal microbiota indicated that dietary supplementation with MBP altered microbial diversity and composition. Specifically, the Sobs, Ace, and Chao indices were significantly lower in the TRE1 group than those in the CON group (p < 0.05). At the phylum level, the relative abundance of Actinobacteriota was significantly higher in the TRE1 and TRE3 groups that in the CON group (p < 0.05). At the genus level, the proportion of WPS-2 in the TRE2 group was significantly higher than that in the CON group (p < 0.05), while the proportion of Clostridia_UCG-014 in the TRE3 group was significantly lower than that in the CON group (p < 0.05). Correlation analysis further revealed that the abundances of Ruminococcus_torques_group and NK4A214_group (phylum Firmicutes) were significantly negatively correlated with daily feed intake, whereas Blautia showed a significant positive correlation with daily feed intake (p < 0.05). Moreover, the abundance of NK4A214_group was significantly negatively correlated with feed efficiency, while the abundance of Olsenella (phylum Actinobacteriota) was significantly positively correlated with feed efficiency (p < 0.05). Discussion:In conclusion, excessive supplementation of MBP (4%) led to thickened intestinal walls, decreased nutrient utilization, and reduced laying rate and egg mass in hens, which may be associated with a reduction in the abundance of beneficial cecal bacteria. Moderate MBP supplementation (2-3%) as a partial substitute for corn had no adverse effects on performance, despite some changes in digestibility and microbiota. This suggests that MBP has potential as an alternative feed ingredient for corn in laying hen diets, but its inclusion level requires careful consideration.
Sojae Semen Praeparatum (SSP) is a traditional food and medicine. This study aimed to elucidate microbial succession during fermentation and its correlations with key metabolites. An absolute quantitative sequencing (AQS) approach was employed, which quantifies microbial copy numbers and accounts for changes in total microbial load across fermentation stages to accurately capture microbial succession. SSP samples from both koji-making and post-fermentation were analyzed using AQS, GC-MS, and physicochemical profiling. Progressive increases were observed in free isoflavones, polyphenols, ammonia nitrogen (AN), total acids, and water-/alcohol-soluble extracts. GC-MS identified 54 volatiles, mainly acids, esters, and amines, showing stage-specific variations. AQS revealed Bacillus and Aspergillus flavus as dominant during koji-making, while Pediococcus and Enterococcus predominated in the post-fermentation stage, representing a new insight into SSP succession. Significant correlations (|r| > 0.8, p < 0.05) were observed between dominant taxa and key components (e.g., isoflavones, polyphenols, AN, and 21 volatiles). Interestingly, this study discusses dual roles of dominant taxa in producing both beneficial components (e.g., isoflavones and polyphenols) and potential risk compounds (e.g., biogenic amines and aflatoxins). These findings highlight AQS as a powerful tool for investigating complex microbial dynamics and provide a basis for optimizing SSP fermentation and quality management.
Medicinal plants and animal-derived proteins represent valuable natural sources of bioactive components with pharmaceutical potential. Whilst some medicinal plants and animal-derived proteins also offer rich sources of anticoagulant bioactive peptides, their development faces multiple challenges: anticoagulant evaluation relies on single-parameter assays with limited reliability, native proteins demonstrate suboptimal activity without enzymatic treatment, and few researchers investigate bioavailable peptides. Our study establishes an innovative framework using the leech as a case study to overcome these barriers. A novel anticoagulant evaluation model was first established with the Critic-G1 weighting method. And we optimized the enzymatically hydrolyzed extracts with high activity using Box–Behnken response surface methodology. Subsequently, the everted gut sac model was implemented to simulate intestinal absorption and screen for absorbable peptide fractions. Furthermore, peptidomics was employed to identify the bioactive peptides. Lastly, we identified the bioactivity using anticoagulation assays. Results indicated that the optimal hydrolysis conditions were achieved with trypsin at 50.48 °C, an enzyme-to-substrate ratio of 6.78%, 7.51 h, and pH of 8.06. The peptide DLRWM was identified through integrated peptidomics and molecular docking approaches, with subsequent activity validation demonstrating its potent anticoagulant effects. This study has successfully identified a novel anticoagulant peptide (DLRWM) with confirmed intestinal absorption properties and provides a template for unlocking the pharmaceutical potential of medicinal animal proteins.
Bacillus velezensis, a member of phylum Firmicutes, is a Gram-positive rod-shaped and endospore-forming bacterium inhabiting diverse environments. The preliminary analysis of the whole genome of Bacillus velezensis ANY11, isolated from bamboo fiber humus in the intestines of Cyrtotrachelus buqueti, revealed that the genome is approximately 3,949,880 bp in length and contains 4265 coding genes. Among these, 12815, 2473, 3193, and 171 genes were annotated in the GO, KEGG, COG, and CAZy databases, respectively. Additionally, the virulence, pathogenicity, and antibiotic resistance of Bacillus velezensis ANY11 were analyzed using PHI, VFDB, and CARD databases. Based on genomic sequencing and gene function analysis, Bacillus velezensis is believed to possess certain disease resistance and the capability to hydrolyze lignocellulose, suggesting its potential role in plant disease prevention and control, as well as in the hydrolysis and reuse of lignocellulose. The bioinformatics from this study may lay a foundation for the production of biofertilizers and biopesticides, as well as for biomass utilization in clean ethanol production.
Hirudinaria manillensis is an important blood-sucking medicinal leech widely distributed in Southeast Asia and southern China. It has been shown to treat cardiovascular and cerebrovascular diseases. Molecular identification technology has developed rapidly, and the results are accurate and stable, which can help us to obtain rich biological information of samples for genetic evolution analysis. This study designed 3 pairs of primers for complete cox 1 gene. After sequencing by PCR products and splicing by SeqMan software, the full-length sequence of cox 1 gene was obtained by annotation of open reading frame (ORFs) on MITOS2 platform. The cox 1 gene with a length of 1536nt was obtained, and the species was identified as Hirudinaria manillensis. The Hd value and FST value of cox 1 gene were 0.858 and 0.10938, which were higher than the Hd value of ITS (0.314) and the FST value close to 0. The nucleotide diversity and GST values of the two genes were very low, and the Tajima’s D values of the neutral test were negative, without statistical significance. The genetic distance calculated by the Kimura-2 parameter method was very low. According to the biological information provided by the samples in this study, the population size of Hirudinaria manillensis in southern China was on the rise, but stable genetic diversity was not formed, and there was no obvious population differentiation and geographical isolation. This may be due to the mature artificial breeding technology in recent years, which enabled Hirudinaria manillensis to migrate from the traditional gathering places and breed in a large number in the new environment and migration site. At present, there are many molecular barcoding data of cox 1 gene in the NCBI database, but there is no complete cox 1 gene sequence as a reference, and the related ITS sequence information is more limited, indicating that there is still a lot of space for gene research of Hirudinaria manillensis.
Levulinic acid (LA) is the main toxic by-product in the production of fuel ethanol, and its large-scale emission adversely affect the ecological environment. In order to effectively remove LA from the liquid waste, microbial degradation methods are adopted but the challenge is that microorganisms cannot fully tolerate LA in the waste. Therefore, it is particularly important to explore the tolerance mechanism of microorganisms to LA. In this study, the whole-genome knockout library scanning and sensitive knockout strain identification were carried out. In addition, subcellular structures such as mitochondria, vacuoles, and endoplasmic reticulum as well as reactive oxygen species (ROS) accumulation were observed under a fluorescence microscopy after stained with fluorescent dyes such as 2'7'-DCF diacetate, Mito Tracker Green FM, Vacuole Membrane Marker MDY-64, and ER-Tracker Red dye. We also performed genomic sequencing on the wild-type strain and knockout strain. Through comparative genomic analysis, it's been found that the LDH1 (YBR204C) gene in Saccharomyces cerevisiae helps promote the clearance of intracellular reactive oxygen species, and the deletion of LDH1 leads to a more-than-two-fold down-regulation of genes related to cell membrane, cell wall, and cell cycle. By measuring the transcriptome and metabolome of the LDH1 knockout strain (ldh1Δ) under LA stress and comparing it with the wild-type strain BY4741, we found that under the condition of LDH1 knockout, the accumulation of NAD+ intermediates would be activated, disrupting normal cell functions and causing cell damage. The LDH1 gene knockout also affects the Methyl Cycle in the cell, which is closely related to the accumulation of ROS. These research results make it possible to create a new genetically modified strain of S. cerevisiae with desired higher tolerance which enhances degradation efficiency and reduces cost.
Abstract Ceracris kiangsu Tsai (C. kiangs) is a kind of forest pest, which can harm nearly 100 kinds of weeds and crops. In this study, based on 314 species distribution points of C. kiangsu which were obtained from Chinese herbaria, literatures and investigation, and data of three future climate scenarios presented by CMIP6, two niche models (Garp, Maxent) were used to predict the suitable area of C. kiangsu in China. The result shows that the main environmental factors affecting the distribution of C. kiangsu are precipitation of driest month (bio14) and min temperature of coldest month (bio6). No matter now and future, the potential distribution areas of C. kiangsu in China are mainly in the south of Qinling–Huaihe River. Under current scenarios, the areas of the total, highly, moderately and poorly suitable of C. kiangsu in China are 160.65 × 104 km2, 31.70 × 104 km2, 60.36 × 104 km2 and 68.59 × 104 km2 respectively. The southern Hubei, western Jiangxi and eastern Hunan are highly-suitable areas. Under SSP1-2.6 and SSP2-4.5 scenarios, both the total suitable area and the highly suitable show a decreasing tread in 2050s. Compared to the 2050s, the total suitable area will coninue to decease in 2090s under SSP1-2.6, while it will increase under SSP2-4.5. The highly suitable area will increase in both scenarios, and the increased percentage under SSP2-4.5 is greater than that under SSP1-2.6. Under SSP5-8.5 scenarios, the total suitable area will increase by 1.83% in the 2050s, and decrease by 1.17% in the 2090s. The highly suitable area in the 2050s and 2090s under this scenarios is larger than under current scenarios. No matter what the scenario, the southern part of Yunnan, the southeast of Sichuan and the southwest of Chongqing will become highly-suitable areas as the climate continues to warm and should be monitored more cosely.
Levulinic acid, a hydrolysis product of lignocellulose, can be metabolized into important compounds in the field of medicine and pesticides by engineered strains of Saccharomyces cerevisiae. Levulinic acid, as an intermediate product widely found in the conversion process of lignocellulosic biomass, has multiple applications. However, its toxicity to Saccharomyces cerevisiae reduces its conversion efficiency, so screening Saccharomyces cerevisiae genes that can tolerate levulinic acid becomes the key. By creating a whole-genome knockout library and bioinformatics analysis, this study used the phenotypic characteristics of cells as the basis for screening and found the HMX1 gene that is highly sensitive to levulinic acid in the oxidative stress pathway. After knocking out HMX1 and treating with levulinic acid, the omics data of the strain revealed that multiple affected pathways, especially the expression of 14 genes related to the cell wall and membrane system, were significantly downregulated. The levels of acetyl-CoA and riboflavin decreased by 1.02-fold and 1.44-fold, respectively, while the content of pantothenic acid increased. These findings indicate that the cell wall-membrane system, as well as the metabolism of acetyl-CoA and riboflavin, are important in improving the resistance of Saccharomyces cerevisiae to levulinic acid. They provide theoretical support for enhancing the tolerance of microorganisms to levulinic acid, which is significant for optimizing the conversion process of lignocellulosic biomass to levulinic acid.
This study examined the phenol degradation capabilities and oxidative stress responses of Candida tropicalis SHC-03, demonstrating its metabolic superiority and resilience compared to Saccharomyces cerevisiae BY4742 in a culture medium with phenol as the sole carbon source. Through comparative growth, transcriptomic, and metabolomic analyses under different phenol concentrations, this study revealed C. tropicalis SHC-03’s specialized adaptations for thriving in phenol as the sole carbon source environments. These include a strategic shift from carbohydrate metabolism to enhanced phenol degradation pathways, highlighted by the significant upregulation of genes for Phenol 2-monoxygenase and Catechol 1,2-dioxygenase. Despite phenol levels reaching 1.8 g/L, C. tropicalis exhibits a robust oxidative stress response, efficiently managing ROS through antioxidative pathways and the upregulation of genes for peroxisomal proteins like PEX2, PEX13, and PMP34. Concurrently, there was significant upregulation of genes associated with membrane components and transmembrane transporters, enhancing the cell’s capacity for substance exchange and signal transduction. Especially, when the phenol concentration was 1.6 g/L and 1.8 g/L, the degradation rates of C. tropicalis towards it were 99.47 and 95.91%, respectively. Conversely, S. cerevisiae BY4742 shows limited metabolic response, with pronounced growth inhibition and lack of phenol degradation. Therefore, our study not only sheds light on the molecular mechanisms underpinning phenol tolerance and degradation in C. tropicalis but also positions this yeast as a promising candidate for environmental and industrial processes aimed at mitigating phenol pollution.
Chicory (Cichorium intybus L., 2n = 18), belonging to the Asteraceae family, exhibits significant edible, medicinal, and pasture values. Moderate research has been performed on identifying Chicory species’ chromosomes using fluorescence in situ hybridization (FISH) and C-banding. Detailed karyotype comparisons with chromosome nomenclature have not yet been performed for Chicory and similar species. In this study, the tandem repeats (TRs) were predicted and mapped to chromosomal regions based on released C. intybus L. ASM2352571 genome assembly v1, and then compared to the genome of Lettuce (Lactuca sativa L.). Nine new oligo probes were then developed and employed for karyotypic investigation of endive, Lettuce, and Chicory mitotic metaphase using non-denaturing FISH (ND-FISH). By combining the conserved oligo probes for 5S rDNA and 18S rDNA with the unique ND-FISH signals of new TR-oligo probes, we can develop a high-resolution standard karyotype for the cultivars of Lettuce and Chicory. The occurrence of chromosome structure variations from the natural population of Chicory and Lettuce was also revealed by ND-FISH with multiple oligo probes. The current observation of the karyotype differences and divergences of Lactuca and Cichorium and the genomic research offers crucial information about the Asteraceae family’s genetic diversity, chromosomal dynamics, and evolutionary routes.
BACKGROUND:Bamboo is an important nontimber forestry product worldwide, while growth, development and geographic distribution of bamboo are often affected by abiotic stresses. Fatty acid desaturases have important roles in regulating plant abiotic stress tolerance, especially low-temperature. However, there is no report on genome-wide of FAD genes in bamboo under abiotic stresses. RESULTS:A toltal of 43 PeFAD genes were identified in moso bamboo genome, which were unevenly located in 17 scaffolds. Phylogenetic analysis indicated that PeFAD genes were divided into 6 groups and ADS/FAD5 group was absence in momo bamboo, and gene structure and histidine-motifs remained highly conserved in each group. The expansion of PeFAD genes was mainly caused by tandem and segmental duplications of SAD/FAB2 group. We also identified 59 types of miRNAs targeting PeFAD genes. RNA-seq data indicated that PeFAD genes were transcribed in various organs/tissues with different degrees, and responded to abiotic stresses and hormone treatments, including cold, salt, drought, SA, ABA, BR, NAA and GA. Co-expression analysis under cold stress showed that PeCBF3 might directly bind the promoter of top cold-responsive PeSLD1 gene that contained LTR cis-element and DRE core element. The qRT-PCR assay also validated the expression pattern of PeSLD1 and its upstream regulatory gene PeCBF3. CONCLUSION:In this study, we performed comprehensive genome-wide survey of PeFAD genes in moso bamboo and analyzed their expression patterns in various tissues and organs, and under abiotic stresses and phytohormones treatment. The qRT-PCR assay validated the cold inducibility of PeSLD1 and PeCBF3. This work showed critical roles of PeFAD genes in abiotic stresses responses. This is the first report on genome-wide analysis of PeFAD genes in moso bamboo, which will provide critical gene resources for molecular breeding of stress-toleranct moso bamboo.
Dof transcription factor family in Cyperus esculentus genome was identified and analyzed using bioinformatics. The analysis results revealed that C.esculentus genome contains 29 Dof genes (CesDof), all of which are located in the nucleus according to subcellular localization prediction. CesDof proteinrs have a range of 124 to 512 amino acids, with most being basic proteins. Their secondary structure was mainly irregular curl. The promoter sequence of CesDof genes contains cis-acting elements that respond to light, drought, hormones, low temperature, and circadian rhythm. Codon preference analysis showed that CesDof genes' codon preference ends in T/A. Collinearity analysis revealed that C.esculentus had three pairs of collinear CesDof genes. Additionally, there were 15 pairs of collinear genes between C.esculentus and Arabidopsis thaliana. The genetic relationship between C.esculentus and Rhynchospora pubera was found to be the closest. Phylogenetic tree analysis revealed that 29 CesDof genes of C.esculentus can be classified into 4 subgroups. Additionally, 144 miRNAs were predicted to target these CesDof genes. Furthermore, protein interaction analysis indicated that 15 Dof proteins in C.esculentus had interactions. The qRT-PCR verification results of drought stress and salt stress treatment experiments showed that most CesDof genes were involved in drought stress and salt stress responses, and the gene expression trends under drought stress and salt stress conditions were consistent. These results lay a theoretical foundation for further studying the molecular functions of Dof gene family in C.esculentus and its molecular mechanisms in regulating the life activities of C.esculentus.
Engineering Saccharomyces cerevisiae for biodegradation and transformation of industrial toxic substances such as catechol (CA) has received widespread attention, but the low tolerance of S. cerevisiae to CA has limited its development. The exploration and modification of genes or pathways related to CA tolerance in S. cerevisiae is an effective way to further improve the utilization efficiency of CA. This study identified 36 genes associated with CA tolerance in S. cerevisiae through genome-wide identification and bioinformatics analysis and the ERG6 knockout strain (ERG6Δ) is the most sensitive to CA. Based on the omics analysis of ERG6Δ under CA stress, it was found that ERG6 knockout affects pathways such as intrinsic component of membrane and pentose phosphate pathway. In addition, the study revealed that 29 genes related to the cell wall-membrane system were up-regulated by more than twice, NADPH and NADP+ were increased by 2.48 and 4.41 times respectively, and spermidine and spermine were increased by 2.85 and 2.14 times, respectively, in ERG6Δ. Overall, the response of cell wall-membrane system, the accumulation of spermidine and NADPH, as well as the increased levels of metabolites in pentose phosphate pathway are important findings in improving the CA resistance. This study provides a theoretical basis for improving the tolerance of strains to CA and reducing the damage caused by CA to the ecological environment and human health.
Objective Diabetic kidney disease (DKD) is a common complication of diabetes mellitus. The pathophysiological changes in platelet function and the hypercoagulable state associated with DKD are closely linked to inflammatory processes. Poecilobdella manillensis (PM), a type of leech known for its anticoagulant and antithrombotic properties, has the potential to modulate the inflammatory response in DKD. This study aims to investigate the effect of freeze-dried powder of PM on improving inflammatory injury in rat glomerular mesangial cells and to explore its underlying mechanism. Methods Lipopolysaccharide (LPS) stimulated HBZY-1 rat mesangial cells to establish an in vitro DKD inflammation model. After the intervention with the water extract of freeze-dried powder of PM (FDPM), cell viability, NO content, and the levels of inflammatory factors such as IL-1β, IL-18, and TNF-α were assessed. Finally, utilizing transcriptomics technology, RT-qPCR, and Western blot methods, the mechanism by which FDPM improves inflammatory injury in rat glomerular mesangial cells was explored and preliminarily validated. Results FDPM effectively enhances cell viability and inhibits the production of NO and related inflammatory factors. Transcriptomic analysis suggests that FDPM may exert these effects by regulating the TXNIP / NLRP3 signaling pathway. The mRNA and protein expressions of TXNIP, NLRP3, and MCP-1 in the model cells were reversed by FDPM. Conclusion FDPM may improve the micro-inflammatory state of DKD and slow the progression of the disease by regulating the TXNIP / NLRP3 signaling pathway. This study provides a scientific basis for the clinical application of PM DKD treatment.
DNA binding with one finger(Dof) gene family is a class of transcription factors which play an important role on plant growth and development. Genome-wide identification results indicated that there were 45 Dof genes(ColDof) in C.oleifera genome. All 45 ColDof proteins were non-transmembrane and non-secretory proteins. Phosphorylation site analysis showed that biological function of ColDof proteins were mainly realized by phosphorylation at serine (Ser) site. The secondary structure of 44 ColDof proteins was dominated by random coil, and only one ColDof protein was dominated by α-helix. ColDof genes’ promoter region contained a variety of cis-acting elements, including light responsive regulators, gibberellin responsive regulators, abscisic acid responsive regulators, auxin responsive regulators and drought induction responsive regulators. The SSR sites analysis showed that the proportion of single nucleotide repeats and the frequency of A/T in ColDof genes were the largest. Non-coding RNA analysis showed that 45 ColDof genes contained 232 miRNAs. Transcription factor binding sites of ColDof genes showed that ColDof genes had 5793 ERF binding sites, 4381 Dof binding sites, 2206 MYB binding sites, 3702 BCR-BPC binding sites. ColDof9, ColDof39 and ColDof44 were expected to have the most TFBSs. The collinearity analysis showed that there were 40 colinear locis between ColDof proteins and AtDof proteins. Phylogenetic analysis showed that ColDof gene family was most closely related to that of Camellia sinensis var. sinensis cv.Biyun and Camellia lanceoleosa. Protein-protein interaction analysis showed that ColDof34, ColDof20, ColDof28, ColDof35, ColDof42 and ColDof26 had the most protein interactions. The transcriptome analysis of C. oleifera seeds showed that 21 ColDof genes were involved in the growth and development process of C. oleifera seeds, and were expressed in 221 C. oleifera varieties. The results of qRT-PCR experiments treated with different concentrations NaCl and PEG6000 solutions indicated that ColDof1, ColDof2, ColDof14 and ColDof36 not only had significant molecular mechanisms for salt stress tolerance, but also significant molecular functions for drought stress tolerance in C. oleifera. The results of this study provide a reference for further understanding of the function of ColDof genes in C.oleifera.
Abstract Background Insect Cytochrome P450 monooxygenase (CYPs or P450s) plays an important role in detoxifying insecticides, causing insect populations to develop resistance. However, the molecular functions of P450 gene family in Cyrtotrachelus buqueti genome are still lacking. Results In this study, 71 CbuP450 genes have been identified. The amino acids length of CbuP450 proteins was between 183 aa ~ 1041 aa. They are proteins with transmembrane domains. The main component of their secondary structure is α-helix and random coils. Phylogenetic analysis showed that C. buqueti and Rhynchophorus ferrugineus were the most closely related. This gene family has 29 high-frequency codons, which tend to use A/T bases and A/T ending codons. Gene expression analysis showed that CbuP450_23 in the female adult may play an important role on high temperature resistance, and CbuP450_17 in the larval may play an important role on low temperature tolerance. CbuP450_10, CbuP450_17, CbuP450_23, CbuP450_10, CbuP450_16, CbuP450_20, CbuP450_23 and CbuP450_ 29 may be related to the regulation of bamboo fiber degradation genes in C. buqueti. Protein interaction analysis indicates that most CbuP450 proteins are mainly divided into three aspects: encoding the biosynthesis of ecdysteroids, participating in the decomposition of synthetic insecticides, metabolizing insect hormones, and participating in the detoxification of compounds. Conclusions We systematically analyzed the gene and protein characteristics, gene expression, and protein interactions of CbuP450 gene family, revealing the key genes involved in the stress response of CbuP450 gene family in the resistance of C. buqueti to high or low temperature stress, and identified the key CbuP450 proteins involved in important life activity metabolism. These results provided a reference for further research on the function of P450 gene family in C. buqueti.
To reveal the molecular function of elongation family of very long chain fatty acids(ELO) protein in Cyrtotrachelus buqueti, we have identified 15 ELO proteins from C.buqueti genome. 15 CbuELO proteins were located on four chromosomes. Their isoelectric points ranged from 9.22 to 9.68, and they were alkaline. These CbuELO proteins were stable and hydrophobic. CbuELO proteins had transmembrane movement, and had multiple phosphorylation sites. The secondary structure of CbuELO proteins was mainly α-helix. A total of 10 conserved motifs were identified in CbuELO protein family. Phylogenetic analysis showed that molecular evolutionary relationships of ELO protein family between C. buqueti and Tribolium castaneum was the closest. Developmental transcriptome analysis indicated that CbuELO10, CbuELO13 and CbuELO02 genes were key enzyme genes that determine the synthesis of very long chain fatty acids in pupae and eggs, CbuELO6 and CbuELO7 were that in the male, and CbuELO8 and CbuELO11 were that in the larva. Transcriptome analysis under different temperature conditions indicated that CbuELO1, CbuELO5, CbuELO12 and CbuELO14 participated in regulating temperature stress responses. Transcriptome analysis at different feeding times showed CbuELO12 gene expression level in all feeding time periods was significant downregulation. The qRT-PCR experiment verified expression level changes of CbuELO gene family under different temperature and feeding time conditions. Protein-protein interaction analysis showed that 9 CbuELO proteins were related to each other, CbuELO1, CbuELO4 and CbuELO12 had more than one interaction relationship. These results lay a theoretical foundation for further studying its molecular function during growth and development of C. buqueti.
IntroductionThis study was conducted to explore the effects of supplementary feeding of bamboo powder on the physical parameters of sows during the perinatal period of 7 days +/- in parturition, including farrow duration, serum biochemical indexes, fecal physicochemical indexes, and microbial flora. MethodsThirty pregnant sows were randomly divided into three groups: the control group was fed a basal diet, TRE1 group and TRE2 group were fed a basal diet supplemented with 30 g d(-1) and 60 g d(-1) bamboo powder, respectively. Multiple parameters of sows and offspring piglets were determined. ResultsThe contents of serum total cholesterol and triglyceride of sows in TRE2 group were significantly lower than those in the control group. The contents of serum malondialdehyde of sows in TRE2 and TRE1 groups were significantly lower than that in control group. The water content of sow feces in TRE2 group was significantly higher than that in control group, and the pH values of sows in TRE2 and TRE1 groups were significantly higher than that in control group. The richness index (Chao) of sow fecal bacterial community in TRE2 group was significantly lower than that of the control group, and the Ace and Sobs indexes tended to be lower than those of the control group. At the phylum level, the relative abundance of Actinobacteriota in the feces of sows in TRE2 group was significantly lower than that of the control group, while that of Fusobacteriota in the feces of suckling piglets in TRE2 group tended to be lower than that of the control group. At the genus level, among the Top10 dominant bacteria, the relative abundance of Tissierella in the feces of sows in TRE2 group was significantly lower than that of the control group while that of Fusobacterium in the feces of suckling piglets in TRE2 group tended to be lower than that of the control group. The relative abundance of Clostridium_sensu_stricto_1, Terrisporobacter, Turicibacter, and Tissierella in the feces of sows in TRE2 group was significantly lower than that of TRE1 group (p < 0.05), while Lactobacillus tended to be higher than that of TRE1 group (p < 0.10). DiscussionThe results suggested that supplementary feeding 60 g d(-1) bamboo powder could increase the water content in the feces of sows, reduce the oxidative damage, and tend to reduce the relative abundance of opportunistic pathogenic Fusobacterium for suckling piglets, while it reduced the fecal microbial diversity of sows.
In this experiment, a ZnO-fibre complex was prepared using the hydrothermal methods of "water solubility," "coupling agent," and "high temperature and high pressure". Binding rate, antibacterial activity, microstructure, and the infrared spectrum were measured using biomimetic digestion, bacterial proliferation tests, and ultra-fine electron microscopes. At first, ZnO-fibre complexes were prepared with different ratios of material and water. They was divided into five groups with ratios of 1:0, 1:4, 1:6, 1:8, and 1:10, respectively. The ZnO-fibre complexes were prepared with different coupling agents on the basis of experiment 1. They were divided into four groups. The ratio for material and water in the control group was 1:0, and in the treatment group, was 1:4. Treatment groups 2 and 3 had 10% guar gum or 10% bamboo fibre polymer composites (BFP) added on the basis of group 1. A ZnO-fibre complex was successfully prepared by adding 10% BFP at a ratio of material:water of 1:4, at a high temperature of 120 °C and a high pressure of 0.3 MPa for 20 min. The ZnO-binding rate reached 99.05%. The zinc oxide may bind to the carbonyl group of bamboo powder and adhere to the surface of and gaps in the bamboo fibre. The growth inhibition rate of Escherichia coli, Salmonella, and Staphylococcus aureus was double that of the common ZnO additive and Zn concentration. It is expected to be used as a slow-release ZnO additive.