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Current work reveals the influence of loading parameters on the crack growth behavior of Zr/Ti/steel composite plate with a crack perpendicular to the interface by using experiment and finite element method. Finite element results reveal that loading parameters significantly affect hysteresis loop of Zr/Ti/steel composite plate, stress amplitude and the plastic strain accumulation degree of subsequent cyclic loading under different loading parameters. In addition, equivalent plastic strain at crack tip, its change rate and distribution are also explored in detail. Furthermore, it is also found that the elastic-plastic mismatch will also affect the plastic evolution, that is, strengthen or weaken the effect of loading parameters.

The purpose of the current study was to explore the immunomodulatory effects of different adjuvants combined with inactivated vaccines under Aeromonas veronii TH0426 infection in crucian carp. This study explored the best conditions for A. veronii as an inactivated vaccine, and included an animal safety test. Furthermore, we expressed the flagellin FlaA of the A. veronii TH0426 strain for use as an adjuvant supplemented in the diet. Crucian carp were fed 12 different experimental diets for 35 days, including the administration of 10 different adjuvants and inactivated vaccine combinations (50% aluminum hydroxide gel and inactivated vaccine combination, and inactivated vaccine with 20%, 30%, or 50% glucan, astragalus polysaccharide or flagellin), inactivated vaccine alone, and PBS control without adjuvant and inactivated vaccine. After the 42 day feeding trials, the fish were challenged with A. veronii TH0426, and the survival rate over 14 days was recorded. In addition, flagellin FlaA can be expressed normally in large amounts. All experimental groups produced higher levels of IgM serum titres than the control group in the different feeding cycles. Moreover, the activity of serum ACP, AKP, SOD, and LZM, and the expression of inflammatory factors were significantly increased in the experimental groups compared with the control group. The results of qRT-PCR analysis showed that the transcription levels of the IL-10, IL-1β, IFN-γ and TNF-α genes in heart, liver, spleen and kidney tissues were significantly enhanced by adjuvant treatment, indicating that the addition of adjuvants can significantly promote the body's inflammatory response. In addition, the phagocytic activity of leukocytes in each adjuvant treated group was significantly enhanced compared to that in the groups without adjuvant. After the A. veronii challenge, the survival rate of all adjuvant-treated groups was significantly higher than that of the control group, and the 50% flagellin adjuvant group had the highest rate of 78.37%. Overall, our findings strongly indicate that adjuvants not only significantly improve the body's immunity, but also exhibit a strong anti-infection ability. Importantly, this work provides a new perspective for the prevention and control of aquaculture diseases.

Aeromonas veronii (A. veronii) is an opportunistic pathogen of fish-human-livestock, which poses a threat to the development of aquaculture. Based on our previous studies on proteomics and genomics, we found out that the aodp gene may be related to the virulence of A. veronii TH0426. However, aodp gene encodes a hypothetical protein with an unknown function, and its role in A. veronii TH0426 is not clear. Here, we first constructed a mutant strain (△-aodp) to investigate the functional role of aodp in A. veronii TH0426. Compared with the wild strain A. veronii TH0426, the growth rate of strain △-aodp was slower and was resistant to neomycin and kanamycin, but sensitive to cephalexin. The swimming and swarming ability of △-aodp strain decreased, and the pathogenicity to mice decreased by 15.84-fold. Besides, the activity of caspase-3 in EPCs infected with △-aodp strain was 1.49-fold lower than that of the wild strain. We examined 20 factors closely related to A. veronii virulence, among them 17 genes were down-regulated as a result of aodp deficiency. This study laid a foundation for further studies on the pathogenesis of A. veronii.

Eukaryotic cells can initiate several distinct self-destruction mechanisms to display essential roles for the homeostasis maintenance, development, and survival of an organism. Pyroptosis, a key response mode in innate immunity, also referred to as caspase-1-dependent proinflammatory programmed necrotic cell death activated by human caspase-1/4/5, or mouse caspase-1/11, plays indispensable roles in response to cytoplasmic insults and immune defense against infectious diseases. These inflammatory caspases are employed by the host to eliminate pathogen infections such as bacteria, viruses, protozoans, and fungi. Gasdermin D requires to be cleaved and activated by these inflammatory caspases to trigger the pyroptosis process. Physiological rupture of cells results in the release of proinflammatory cytokines, the alarmins IL-1β and IL-18, symbolizing the inflammatory potential of pyroptosis. Moreover, long noncoding RNAs play direct or indirect roles in the upstream of the pyroptosis trigger pathway. Here, we review in detail recently acquired insights into the central roles of inflammatory caspases, inflammasomes, and pyroptosis, as well as the crosstalk between pyroptosis and long noncoding RNAs in mediating infection immunity and pathogen clearance.

Seven image-based methods to determine mixing uniformity in inline mixers by image processing were compared using viscous pesticides mixtures with gradually increasing uniformity. After obtaining initial optimal methods and calculation parameters, they were further evaluated by a jet mixer and a layered mixer. Results showed that quantitative descriptions for uniformity inside the tube by grey level co-occurrence matrix (GLCM) and concentration difference within time sequence (CVT) failed to reflect accurately the changing-trends in uniformity. A histogram information entropy (HIE)-based method tended to underestimate changes, while coefficient of variance based on each single frame (CVS) which calculated differences in pixel values underestimated it when mixtures showed stratification, indicating that both methods were not suitable. However, the accuracy of methods using explicit features such as second-order moments of grey-scale histogram (HSM) and optimised area-weighted uniformity index (OAU), as well as implicit methods such as principal components analysis (PCA) were suitable, and the normalised index > 80% can be suitable because no non-scattered pesticide remained. Single-viewing images were adequate for evaluating the uniformity produced by mixers although small errors between horizontal and vertical-viewing images existed. During evaluations by mixers, OAU and PCA proved to be more advantageous than HSM because these methods do not only rely on pixel probability distributions of grey-scale histograms, but the positional information of the pesticide distribution, even though the HSM is more time-efficient than them.

The injection of pesticide directly into spray nozzles results in concentration variations, both periodic discontinuities and pulses, caused by the use of injection pumps and solenoid valves. Such variations can lead to inaccurate application of pesticide. To avoid using long pipelines, long lag times and inaccurate application, efficient mixers are required to shorten the distance between the injection point and the spray nozzle. To analyse concentration variations and periodic pulses, a set of methods based on image processing were proposed and were tested using three inline mixers, a jet mixer (A), a layered mixer (B) which was specially designed, and simplified layered-jet mixer (C) which was developed as combination of mixer A and some simplified structures of mixer B. Results showed the characteristics of concentration variability for three mixers were generally different. The application conditions, carrier flow rate, mixing ratio, and injection frequency, need to be comprehensively considered to achieve improved concentration consistency and improve spraying accuracy. Concentration variability was the most obvious for mixer A with the average Coefficient of Variation (CV) as high as 4.12%. Using auto-correlation and complexity analysis, the concentration series exhibited strong periodicity and relatively lower complexity. Concentration variability for mixer B was less prominent with an average CV value 1.25% and concentration periodicity sometimes being indeterminable. Mixer C showed moderate concentration variability (CV 2.26%) although pulsation was still detectable. Mixers with similar structures to mixer C could be applied for direct nozzle injection systems to improve the spraying accuracy.

Aeromonas veronii is a comorbid pathogen that can infect humans, and animals including various aquatic organisms. In recent years, an increasing number of cases of A. veronii infection has been reported, indicating serious risks. This bacterium not only threatens public health and safety but also causes considerable economic loss in the aquaculture industry. Currently, some understanding of the pathogenic mechanism of A. veronii has been obtained. In this study, we first constructed the A. veronii TH0426 fis gene deletion strain Δfis and the complementation strain C-fis through homologous recombination technology. The results showed that the adhesion and invasion ability of the Δfis strain towards Epithelioma papulosum cyprini (EPC) cells and the cytotoxicity were 3.8-fold and 1.38-fold lower, respectively, than those of the wild-type strain. In the zebrafish infection model, the lethality of the deleted strain is 3-fold that of the wild strain. In addition, the bacterial load of the deletion strain Δfis in crucian carp was significantly lower than the wild-type strain, and the load decreased with time. In summary, deletion of the fis gene led to a decrease in the virulence of A. veronii. Our research results showed that the deletion of the fis gene significantly reduces the virulence and adhesion ability of A. veronii TH0426. Therefore, the fis gene plays a vital role in the pathogenesis of A. veronii TH0426. This preliminary study of the function of the fis gene in A. veronii will help researchers further understand the pathogenic mechanism of A. veronii.

Aeromonas veronii is a serious pathogen which can infect mammals and aquatic organisms and causes irreparable damage to fish aquaculture. It has been demonstrated that adhesion to host surface and cells is the initial step in bacterial pathogenesis. Previous study found that bacterial weaken motility probably caused by the absence of flagellar related genes. In this study, we generated the aha deletion and complementary strains and found that two strains can be stably inherited for more than 50 generations. No significant change was found in the growth of mutant △aha. But the ability of biofilm formation, the adhesion and invasion to EPC cells significantly decreased for 3.7-fold and 2.3-fold respectively. Due to aha gene deletion, the stability of A. veronii flagellar was severely declined and the mutant △aha with no mobility. Compared with the wild-type TH0426, the pathogenicity of A. veroniiaha-deleted strain to zebrafish and mice reduced significantly and virulence attenuated severely. Cytotoxicity experiment also proved that mutant △aha showed much weaker virulence at the same time infection. The consequences declared that the stability of flagellar decreased severely with porin missing and lost the motility. Porin regulated by aha gene is essential for the adhesion and virulence of A. veronii. Thence, the mutant △aha of A. veronii provides an important tool for further concentration on the pathogenic mechanism of A. veronii.

Aeromonas veronii (A. veronii, AV) strains are emerging zoonotic and aquatic pathogens, yet we know very little about their genomics. This study aims to utilize comparative genomics to investigate the intraspecific genetic diversity, differences in virulence factors and evolutionary mechanisms of A. veronii strains from diverse sources and to fundamentally demonstrate their pathogenic mechanisms. We conducted comparative genomics analysis of 39 A. veronii strains from different sources and found that 1993 core genes are shared by these strains and that these shared core genes may be necessary to maintain the basic characteristics of A. veronii. Additionally, phylogenetic relationship analysis based on these shared genes revealed that a distant relationship between the AMC34 strain and the other 38 strains but that, the genetic relationship among the 38 strains is relatively close, indicating that AMC34 may not belong to A. veronii. Furthermore, analysis of shared core genes and average nucleotide identity (ANI) values showed no obvious correlation with the location of A. veronii isolation and genetic relationship. Our research indicates the evolutionary mechanism of A. veronii from different sources and provides new insights for a deeper understanding of its pathogenic mechanism.

Additional file 1: Figure S1. Confirmation of the success and genetic stability of mutant strain ∆dotU and complemented strain C-dotU. (a) PCR detection of the mutant strain ∆dotU, M: DL5000 Marker; 1: the mutant strain ∆dotU; 2: wild type TH0426. (b) PCR detection of the complemented strain C-dotU, M: DL2000 Marker; 1: C-dotU; 2: TH0426. (c) Genetic stability of the partial deletion strain ∆dotU, M: DL5000 Marker; 1–10: ∆dotU; 11–12: TH0426; 13: control group. (d) Genetic stability of the partial complemented strain C-dotU, M: DL2000 Marker; 1–10: C-dotU; 11–12: TH0426; 13: control group. Figure S2. Flagellum straining and light microscopy observation of the three strains (the wild-type, ∆dotU and C-dotU). Light microscopy images of parental strain (A), ∆dotU (B) and C-dotU (C). Magnifications, 1000 × (A, B and C).

Aeromonas veronii is one of the main pathogens causing freshwater fish sepsis and ulcer syndrome. This bacterium has caused serious economic losses in the aquaculture industry worldwide, and it has become an important zoonotic and aquatic agent. However, little is known about the molecular mechanism of pathogenesis of A. veronii. In this study, we first constructed an unmarked mutant strain (&Delta, preA) by generating an in-frame deletion of the preA gene, which encodes a periplasmic binding protein, to investigate its role in A. veronii TH0426. Our results showed that the motility and biofilm formation ability of &Delta, preA were similar to those of the wild-type strain. However, the adhesion and invasion ability in epithelioma papulosum cyprini (EPC) cells were significantly enhanced (2.0-fold). Furthermore, the median lethal dose (LD50) of &Delta, preA was 7.6-fold higher than that of the wild-type strain, which illustrates that the virulence of the mutant was significantly enhanced. This finding is also supported by the cytotoxicity test results, which showed that the toxicity of &Delta, preA to EPC cells was enhanced 1.3-fold relative to the wild type. Conversely, tolerance test results showed that oxidative stress resistance of &Delta, preA decreased 5.9-fold compared to with the wild-type strain. The results suggest that preA may negatively regulate the virulence of A. veronii TH0426 through the regulation of resistance to oxidative stress. These insights will help to further elucidate the function of preA and understand the pathogenesis of A. veronii.

Aeromonas veronii is one of the main pathogens causing freshwater fish sepsis and ulcer syndrome. This bacterium has caused serious economic losses in the aquaculture industry worldwide, and it has become an important zoonotic and aquatic agent. However, little is known about the molecular mechanism of pathogenesis of A. veronii. In this study, we first constructed an unmarked mutant strain (ΔpreA) by generating an in-frame deletion of the preA gene, which encodes a periplasmic binding protein, to investigate its role in A. veronii TH0426. Our results showed that the motility and biofilm formation ability of ΔpreA were similar to those of the wild-type strain. However, the adhesion and invasion ability in epithelioma papulosum cyprini (EPC) cells were significantly enhanced (2.0-fold). Furthermore, the median lethal dose (LD50) of ΔpreA was 7.6-fold higher than that of the wild-type strain, which illustrates that the virulence of the mutant was significantly enhanced. This finding is also supported by the cytotoxicity test results, which showed that the toxicity of ΔpreA to EPC cells was enhanced 1.3-fold relative to the wild type. Conversely, tolerance test results showed that oxidative stress resistance of ΔpreA decreased 5.9-fold compared to with the wild-type strain. The results suggest that preA may negatively regulate the virulence of A. veronii TH0426 through the regulation of resistance to oxidative stress. These insights will help to further elucidate the function of preA and understand the pathogenesis of A. veronii.

A case study of a widened highway embankment is presented, in which prestressed tubular concrete (PTC) piles and geogrids were used to reinforce the embankment and to reduce construction ti...

Aeromonas veronii is a pathogenic gram-negative bacterium, which infects a variety of animals and results in mass mortality. The stalled-ribosome rescues are reported to ensure viability and virulence under stress conditions, of which primarily include trans-translation and alternative ribosome-rescue factor A (ArfA) in A. veronii. For identification of specific peptides that interact and inhibit the stalled-ribosome rescues, peptide aptamer library (pTRG-SN-peptides) was constructed using pTRG as vector and Staphylococcus aureus nuclease (SN) as scaffold protein, in which 16 random amino acids were introduced to form an exposed surface loop. In the meantime both Small Protein B (SmpB) which acts as one of the key components in trans-translation, and alternative ribosome-rescue factor A (ArfA) were inserted to pBT to constitute pBT-SmpB and pBT-ArfA, respectively. The peptide aptamer PA-2 was selected from pTRG-SN-peptides by bacterial two-hybrid system (B2H) employing pBT-SmpB or pBT-ArfA as baits. The conserved sites G133K134 and D138K139R140 of C-terminal SmpB were identified by interacting with N-terminal SN, and concurrently the residue K62 of ArfA was recognized by interacting with the surface loop of the specific peptide aptamer PA-2. The expression plasmids pN-SN or pN-PA-2, which combined the duplication origin of pRE112 with the neokanamycin promoter expressing SN or PA-2, were created and transformed into A. veronii C4, separately. The engineered A. veronii C4 which endowing SN or PA-2 expression impaired growth capabilities under stress conditions including temperatures, sucrose, glucose, potassium chloride (KCl) and antibiotics, and the stress-related genes rpoS and nhaP were down-regulated significantly by Quantitative Real-time PCR (qRT-PCR) when treating in 2.0% KCl. Thus，the engineered A. veronii C4 conferring PA-2 expression might be potentially attenuated vaccine, and also the peptide aptamer PA-2 could develop as anti-microbial drugs targeted to the ribosome rescued factors in A. veronii.

Earlier studies reveal that Small Protein B (SmpB), a class of well-conserved tmRNA-binding proteins, is essential for the trans-translation process, which functions as a system for translation surveillance and ribosome rescue. Here, we report a previously unrecognized mechanism by which SmpB alone positively regulates the expression of a sensor kinase, BvgS, in Aeromonas veronii. A reporter plasmid was constructed in which the promoter of bvgS was used to control the expression of the enhanced green fluorescent protein (eGFP) gene. When the reporter plasmid was co-transformed with a SmpB expression construct into E. coli, the relative fluorescence intensity increased about 3-fold. Transformation with a truncated form of smpB gene showed that the C-terminus had little effect, while N-terminus unexpectedly increased eGFP production. Next, a series of SmpB mutants were generated by site-directed mutagenesis. When the mutants SmpB (G11S) or SmpB (E32AG) was used in the experiment, eGFP expression dropped significantly compared with that of wild type SmpB. Further，purified SmpB was shown to bind the promoter regions of bvgS in the agarose gel retardation assay. Quantitative RT-PCR analysis showed that eGFP transcript levels increased approximately 25-fold in the presence of SmpB. Likewise, bvgS transcripts decreased significantly in smpB knockout A. veronii. Similar to BvgS inhibition, smpB knockout in A. veronii displayed a reduced capability in salt tolerance. Collectively, the data presented here will facilitate a deeper understanding of SmpB-mediated regulatory circuits as a transcriptional factor in A. veronii.