Changes of intestinal barrier in the process of intestinal inflammation induced by Aeromonas hydrophila in snakehead (Channa argus).

Bacterial intestinal inflammation frequently occurs in cultured fish. Nevertheless, research on intestinal barrier dysfunction in the process of intestinal inflammation is deficient. In this study, we explored the changes of intestinal inflammation induced by Aeromonas hydrophila (A. hydrophila) in snakehead and the relationship between intestinal barrier and inflammation. Snakehead [(13.05 ± 2.39) g] were infected via anus with A. hydrophila. Specimens were collected for analysis at 0, 1, 3, 7 and 21 d post-injection. The results showed that with the increase of exposure time, the hindgut underwent stages of normal function, damage, damage deterioration, repair and recovery. Relative to 0 d, the levels of IL-1ß and TNF-α in serum, and the expression of nod1, tlr1, tlr5, nf-κb, tnf-α and il-1ß in intestine were significantly increased, and showed an upward then downward pattern over time. However, the expression of tlr2 and il-10 were markedly decreased, and showed the opposite trend. In addition, with the development of intestinal inflammation, the diversity and richness of species, and the levels of phylum and genus in intestine were obviously altered. The levels of trypsin, LPS, AMS, T-SOD, CAT, GPx, AKP, LZM and C3 in intestine were markedly reduced, and displayed a trend of first decreasing and then rebounding. The ultrastructure observation showed that the microvilli and tight junction structure of intestinal epithelial cells experienced normal function initially, then damage, and finally recovery over time. The expression of claudin-3 and zo-1 in intestine were significantly decreased, and showed a trend of first decreasing and then rebounding. Conversely, the expression of mhc-i, igm, igt and pigr in intestine were markedly increased, and displayed a trend of increasing first and then decreasing. The above results revealed the changes in intestinal barrier during the occurrence and development of intestinal inflammation, which provided a theoretical basis for explaining the relationship between the two.

Incidence and clinical analysis of asymptomatic intracranial hemorrhage in neonates with cerebral hypoxic-ischaemic risk based on multisequence MR images.

The incidence and clinical distribution of intracranial haemorrhage (ICH) in neonates at risk of cerebral hypoxia-ischaemia have not been reported in specific studies. Based on conventional magnetic resonance imaging (MRI) versus susceptibility weighted imaging (SWI), this study aimed to analyse the occurrence of asymptomatic ICH in newborns with or without risk of cerebral hypoxia-ischaemia and to accumulate objective data for clinical evaluations of high-risk neonates and corresponding response strategies. 317 newborns were included. MRI revealed that the overall incidence of ICH was 59.31%. The most common subtype was intracranial extracerebral haemorrhage (ICECH) which included subarachnoid haemorrhage (SAH) and subdural haemorrhage (SDH). ICECH accounted for 92.02% of ICH. The positive detection rate of ICECH by SWI was significantly higher than that by T1WI. The incidence of total ICH, ICECH and SAH was greater among children who were delivered vaginally than among those who underwent caesarean delivery. Asymptomatic neonatal ICH may be a common complication of the neonatal birth process, and SWI may improve the detection rate. Transvaginal delivery and a weight greater than 2500 g were associated with a high incidence of ICECH in neonates. The impact of neonatal cerebral hypoxia-ischaemia risk factors on the occurrence of asymptomatic ICH may be negligible.

Poly (I:C)-Induced microRNA-30b-5p Negatively Regulates the JAK/STAT Signaling Pathway to Mediate the Antiviral Immune Response in Silver Carp (Hypophthalmichthys molitrix) via Targeting CRFB5.

In aquaculture, viral diseases pose a significant threat and can lead to substantial economic losses. The primary defense against viral invasion is the innate immune system, with interferons (IFNs) playing a crucial role in mediating the immune response. With advancements in molecular biology, the role of non-coding RNA (ncRNA), particularly microRNAs (miRNAs), in gene expression has gained increasing attention. While the function of miRNAs in regulating the host immune response has been extensively studied, research on their immunomodulatory effects in teleost fish, including silver carp (Hyphthalmichthys molitrix), is limited. Therefore, this research aimed to investigate the immunomodulatory role of microRNA-30b-5p (miR-30b-5p) in the antiviral immune response of silver carp (Hypophthalmichthys molitrix) by targeting cytokine receptor family B5 (CRFB5) via the JAK/STAT signaling pathway. In this study, silver carp were stimulated with polyinosinic-polycytidylic acid (poly (I:C)), resulting in the identification of an up-regulated miRNA (miR-30b-5p). Through a dual luciferase assay, it was demonstrated that CRFB5, a receptor shared by fish type I interferon, is a novel target of miR-30b-5p. Furthermore, it was found that miR-30b-5p can suppress post-transcriptional CRFB5 expression. Importantly, this study revealed for the first time that miR-30b-5p negatively regulates the JAK/STAT signaling pathway, thereby mediating the antiviral immune response in silver carp by targeting CRFB5 and maintaining immune system stability. These findings not only contribute to the understanding of how miRNAs act as negative feedback regulators in teleost fish antiviral immunity but also suggest their potential therapeutic measures to prevent an excessive immune response.

Downregulation of miR-1388 Regulates the Expression of Antiviral Genes via Tumor Necrosis Factor Receptor (TNFR)-Associated Factor 3 Targeting Following poly(I:C) Stimulation in Silver Carp (Hypophthalmichthys molitrix).

MicroRNAs (miRNAs) are highly conserved endogenous single-stranded non-coding RNA molecules that play a crucial role in regulating gene expression to maintain normal physiological functions in fish. Nevertheless, the specific physiological role of miRNAs in lower vertebrates, particularly in comparison to mammals, remains elusive. Additionally, the mechanisms underlying the control of antiviral responses triggered by viral stimulation in fish are still not fully understood. In this study, we investigated the regulatory impact of miR-1388 on the signaling pathway mediated by IFN regulatory factor 3 (IRF3). Our findings revealed that following stimulation with the viral analog poly(I:C), the expression of miR-1388 was significantly upregulated in primary immune tissues and macrophages. Through a dual luciferase reporter assay, we corroborated a direct targeting relationship between miR-1388 and tumor necrosis factor receptor (TNFR)-associated factor 3 (TRAF3). Furthermore, our study demonstrated a distinct negative post-transcriptional correlation between miR-1388 and TRAF3. We observed a significant negative post-transcriptional regulatory association between miR-1388 and the levels of antiviral genes following poly(I:C) stimulation. Utilizing reporter plasmids, we elucidated the role of miR-1388 in the antiviral signaling pathway activated by TRAF3. By intervening with siRNA-TRAF3, we validated that miR-1388 regulates the expression of antiviral genes and the production of type I interferons (IFN-Is) through its interaction with TRAF3. Collectively, our experiments highlight the regulatory influence of miR-1388 on the IRF3-mediated signaling pathway by targeting TRAF3 post poly(I:C) stimulation. These findings provide compelling evidence for enhancing our understanding of the mechanisms through which fish miRNAs participate in immune responses.

Population characteristics of pathogenic Escherichia coli in puerperal metritis of dairy cows in Ningxia region of China: a systemic taxa distribution of virulence factors and drug resistance genes.

Escherichia coli (E. coli) is closely associated with the occurrence of puerperal metritis in dairy cows. E. coli carries some the virulence and multi-drug resistant genes, which pose a serious threat to the health of postpartum cows. In this study, E. coli was isolated and identified from the uterine contents of postpartum cows with puerperal metritis in the Ningxia region of China, and its phylogenetic subgroups were determined. Meanwhile, virulence and drug resistance genes carried by E. coli and drug sensitivity were detected, and the characteristics of virulence and drug resistance genes distribution in E. coli phylogroups were further analyzed. The results showed that the isolation rate of E. coli in puerperal metritis samples was 95.2%. E. coli was mainly divided into phylogroups B2 and D, followed by groups A and B1, and was more connected to O157:H7, O169:H4, and ECC-1470 type strains. The virulence genes were mainly dominated by ompF (100%), traT (100%), fimH (97%), papC (96%), csgA (95%), Ang43 (93.9%), and ompC (93%), and the resistance genes were dominated by TEM (99%), tetA (71.7%), aac(3)II (66.7%), and cmlA (53.5%). Additionally, it was observed that the virulence and resistance gene phenotypes could be divided into two subgroups, with subgroup B2 and D having the highest distributions. Drug sensitivity tests also revealed that the E. coli was most sensitive to the fluoroquinolones enrofloxacin, followed by macrolides, aminoglycosides, tetracyclines, ß-lactams, peptides and sulfonamides, and least sensitive to lincosamides. These results imply that pathogenic E. coli, which induces puerperal metritis of dairy cows in the Ningxia region of China, primarily belongs to the group B2 and D, contains multiple virulence and drug resistance genes, Moreover, E. coli has evolved resistance to several drugs including penicillin, lincomycin, cotrimoxazole, and streptomycin. It will offer specific guidelines reference for the prevention and treatment of puerperal metritis in dairy cows with E. coli infections in the Ningxia region of China.

Quercetin targets SarA of methicillin-resistant Staphylococcus aureus to mitigate biofilm formation.

IMPORTANCE: Anti-biofilm is an important strategy against Staphylococcus aureus chronic infection. SarA is a positive regulator of biofilm formation in S. aureus. In this study, we identified the SarA inhibitor quercetin using computer simulation screening. Previous studies have shown that quercetin inhibits biofilm; however, the underlying mechanism remains unknown. This study revealed the inhibitory effect of quercetin on the SarA protein. We also isolated the SarA protein and confirmed its interaction with quercetin in vitro. Besides, the inhibitory effect of quercetin on the transcription and translation levels of the SarA protein was also determined. The effects of quercetin on S. aureus biofilm inhibition and biofilm components were consistent with the changes in the transcription level of biofilm-related genes regulated by SarA. In summary, our study revealed the mechanism by which quercetin affects biofilm formation by inhibiting the transcriptional regulator SarA of S. aureus.

Global Prevalence and Hemagglutinin Evolution of H7N9 Avian Influenza Viruses from 2013 to 2022.

H7N9 avian influenza viruses have caused severe harm to the global aquaculture industry and human health. For further understanding of the characteristics of prevalence and hemagglutinin evolution of H7N9 avian influenza viruses, we generated the global epidemic map of H7N9 viruses from 2013 to 2022, constructed a phylogenetic tree, predicted the glycosylation sites and compared the selection pressure of the hemagglutinin. The results showed that although H7N9 avian influenza appeared sporadically in other regions worldwide, China had concentrated outbreaks from 2013 to 2017. The hemagglutinin genes were classified into six distinct lineages: A, B, C, D, E and F. After 2019, H7N9 viruses from the lineages B, E and F persisted, with the lineage B being the dominant. The hemagglutinin of highly pathogenic viruses in the B lineage has an additional predicted glycosylation site, which may account for their persistent pandemic, and is under more positive selection pressure. The most recent ancestor of the H7N9 avian influenza viruses originated in September 1991. The continuous evolution of hemagglutinin has led to an increase in virus pathogenicity in both poultry and humans, and sustained human-to-human transmission. This study provides a theoretical basis for better prediction and control of H7N9 avian influenza.

Beneficial effects of indigenous Bacillus spp. on growth, antioxidants, immunity and disease resistance of Rhynchocypris lagowskii.

This study aimed to investigate the effect of Bacillus aryabhattai (LSG3-7) and Bacillus mojavensis (LSG3-8) on growth performance, antioxidant capacity, and immune response in Rhynchocypris lagowskii (Dybowski, 1869), at the trial and challenge periods. A 630 healthy fish (10.76 ± 0.05) were randomly divided into six groups: control group (D1) was fed the basal diet, D2 and D3 were supplemented with LSG 3-7 and LSG3-8 (1 × 108 CFU/g) for both of them, whereas D4 was supplemented with a mixture of both bacteria (0.5 × 108 CFU/g each), and D5 was supplemented with LSG3-7 0.75 × 108 CFU/g + LSG3-8 0.25 × 108 CFU/g, and D6 supplemented with LSG3-7 0.25 × 108 CFU/g + LSG3-8 0.75 × 108 CFU/g. After the trial, Aeromonas hydrophila was used in a challenge test for 14 days. Treatments showed significant differences (p < 0.05) in growth performance and antioxidant capacity (CAT, CuZn-SOD, GPX) in the liver and intestine compared to the control. The antioxidant-related genes CAT, CuZn-SOD, GPX, and Nrf2 in the liver and intestine showed upregulation compared with the control group. Serum IgM, LZM, C3, C4, and AKP showed a favorable superiority (p < 0.05) in treatments (D2 - D6) at the trial and challenge test compared to controls. In parallel, immune-related genes (IgM, NF-κB, TLR-1, TLR-2, and MyD88) showed an up-regulated level (p < 0.05) in treatments (D2 - D6) compared to the control. In addition, pro-inflammatory cytokines (IL-1, TNF-α) showed a downregulated level in treatments (D2 - D6). After the challenge test, the immune-related genes in the liver and muscle showed an up-regulated level in treatments compared to the controls. The survival rate showed a significant increase (p < 0.05) in the treatment groups (D2 - D6) compared to the control. Overall, individuals and the bacterial mixture of B. aryabhattai and B. mojavensis could improve the growth performance, antioxidant capacity, immune capacity, and survival rate of R. lagowskii and prevent side effects of A. hydrophila. However, B. mojavensis showed a slight improvement compared to B. aryabhattai without a significant difference between them.

High-precision simultaneous measurement of 187Os, 186Os, and 184Os using 1013 Ω amplifiers and CDDs on NTIMS.

High-precision measurements of 184Os/188Os, 186Os/188Os, and 187Os/188Os ratios are significant in the fields of geochemistry and cosmochemistry. However, no high-precision measurement technique exists for simultaneously obtaining all three ratios using a static method for samples with an Os content of <1 ng or 186OsO3‾ and 187OsO3‾ ion-beam intensities of <150 mV. This greatly limits research on rare samples with small sample sizes or low Os contents, such as Lunar, Martian, or old Earth samples. This paper reports a static method, which could achieve the simultaneous measurement of 9 Faraday cups (FCs) with high-signal/noise ratio 1012 Ω amplifiers and 1013 Ω amplifiers and two compact discrete dynodes (CDDs). By analyzing two calibration solutions, a precision value of less than 3‰ (2RSD) could be achieved for 184Os/188Os ratios, even if the 184OsO3‾ intensity was as low as 1000 cps. The precision values for the 186Os/188Os and 187Os/188Os ratios were similar and could be better than 0.066‰ (2RSD) when the intensities of 186OsO3‾ and 187OsO3‾ were greater than 30 mV, which can be obtained with conventional 1011 Ω amplifiers only at signals larger than 150 mV. Three geological reference materials were used in this study. The precision values of 184Os/188Os, 186Os/188Os, and 187Os/188Os ratios reached 2, 0.061, and 0.050‰ (2RSD), respectively, when the maximum Os amount was approximately 12 ng, and 87, 15, and 10‰ (2RSD), respectively, when the maximum Os amount was as low as approximately 66 pg. Additionally, our results show that changes in the volume of oxygen added during measurement can cause significant variations in the oxygen isotope composition. This static measurement method not only avoids the nonlinear signal change of the instrument during the analysis process but also realizes the accurate removal of the oxygen isobaric interference in the run, which could improve the precision of 184Os/188Os, 186Os/188Os, and 187Os/188Os ratios for small-size/low-signal samples.

Study on the inhibitory effect of quercetin combined with gentamicin on the formation of Pseudomonas aeruginosa and its bioenvelope.

OBJECTIVE: The potential effects of quercetin and gentamicin combination on the bacteriostatic activity and biofilm formation of Pseudomonas aeruginosa (PA) were examined, and the findings provided a theoretical basis for the development of quercetin as a new biofilm inhibitor. METHODS: The minimum inhibitory concentration (MIC) of eight PAs was determined by microdilution method and the partial inhibitory concentration index (FICI) of the combined drug was analyzed by micro-dilution method. Thereafter, the lowest film inhibitory concentration (MBIC) of quercetin and gentamicin alone and in combination was evaluated by crystal violet staining. Finally, scanning electron microscopy (SEM) and laser confocal microscopy (CLSM) were used to decipher the inhibitory effect of the combination on biofilm formation. OUTCOME: The antibacterial activity of quercetin alone was relatively weak, but after combination with gentamicin, the antibacterial activity was significantly enhanced, as evident by FICI of 0.28 and 0.53 and manifested as synergistic or additive effect, which indicated that quercetin can enhance gentamicin antibacterial activity. The results of crystal violet staining revealed that quercetin and gentamicin alone exhibited a similar biofilm formation inhibitory effect, but the inhibitory effect was substantially weaker, and the antibiofilm activity was stronger and exhibited a dose-dependent response after the combination of the two with 1/2FICI. The results of scanning electron microscopy and laser confocal microscopy also showed that the treatment of PA biofilm after combining quercetin and gentamicin with 1/2FICI could completely destroy the spatial structure of the complete biofilm, significantly reduce the thickness of bacteria, and markedly reduce the proportion of viable bacteria in the membrane. CONCLUSION: The combination of quercetin and gentamicin can effectively inhibit the formation of PA as well as its biofilm, and exhibit synergistic and additive effects.

Impact of cell-free supernatant of lactic acid bacteria on Staphylococcus aureus biofilm and its metabolites.

Introduction: A safe bio-preservative agent, lactic acid bacteria (LAB) can inhibit the growth of pathogenic bacteria and spoilage organisms. Its cell-free supernatant (LAB-CFS), which is rich in bioactive compounds, is what makes LAB antibacterial work. Methods: This study focused on the changes in biofilm activity and related metabolic pathways of S. aureus treated with lactic acid bacteria planktonic CFS (LAB-pk-CFS) and biofilm state (LAB-bf-CFS). Results: The findings demonstrated that the LAB-CFS treatment considerably slowed Staphylococcus aureus (S. aureus) growth and prevented it from forming biofilms. Additionally, it inhibits the physiological traits of the S. aureus biofilm, including hydrophobicity, motility, eDNA, and PIA associated to the biofilm. The metabolites of S. aureus biofilm treated with LAB-CFS were greater in the LAB-bf-CFS than they were in the LAB-pk-CFS, according to metabolomics studies. Important metabolic pathways such amino acids and carbohydrates metabolism were among the most noticeably altered metabolic pathways. Discussion: These findings show that LAB-CFS has a strong potential to combat S. aureus infections.

Effect of preoperative pupil offset on corneal higher-order aberrations after femtosecond laser-assisted in situ keratomileusis.

BACKGROUND: This study aimed to investigate the relationship between multiple higher-order aberrations (HOAs) subgroups and pupil offset, as well as to analyze the factors affecting postoperative corneal HOAs in patients with different degrees of refractive errors. METHODS: We enrolled 160 patients (316 eyes) aged ≥ 18 years who had undergone femtosecond laser-assisted in situ keratomileusis (FS-LASIK) treatment. Based on the relationship between the preoperative pupil offset and the postoperative ΔHOAs, all patients were divided into two groups: group I (pupil offset ≤ 0.20 mm) and group II (pupil offset > 0.20 mm). All of the eyes had low to high myopia with or without astigmatism (manifest refraction spherical equivalent (MRSE) < -10.00 D). Uncorrected distance visual acuity, corrected distance visual acuity, MRSE, pupil offset, central corneal thickness, corneal HOAs, vertical coma (Z3-1), horizontal coma (Z31), spherical aberration (Z40), trefoil 0° (Z33), and trefoil 30° (Z3-3) over a 6 mm diameter central corneal zone diameter were evaluated preoperatively and at 1 and 3 months postoperatively. RESULTS: Our result revealed significant differences in postoperative corneal total root mean square (RMS) HOAs, RMS vertical coma, RMS horizontal coma, RMS spherical aberration, and RMS trefoil 30° between group I and group II. ΔMRSE was found to be an effective factor for ΔRMS HOAs (R2 = 0.383), ΔRMS horizontal coma (R2 = 0.205), and ΔRMS spherical aberration (R2 = 0.397). In group II, multiple linear regression analysis revealed a significant correlation between preoperative pupillary offset and Δtotal RMS HOAs (R2 = 0.461), ΔRMS horizontal coma (R2 = 0.040), and ΔRMS trefoil 30°(R2 = 0.089). The ΔRMS vertical coma effect factor is the Y-component, and the factor influencing ΔRMS spherical aberration was ΔMRSE (R2 = 0.256). CONCLUSION: A small pupil offset was associated with a lower induction of postoperative corneal HOAs. Efforts to optimize centration are critical for improving surgical outcomes in patients with FS-LASIK.

Characterization of Bacillus spp. isolated from the intestines of Rhynchocypris lagowskii as a potential probiotic and their effects on fish pathogens.

Probiotics sourced from fish intestinal microbiota have a merit over other bacterial sources due to colonization ability and effective time. This study aimed to evaluate the bacilli isolated from the Rhynchocypris lagowskii intestines and their validity as a probiotic. Three isolates were selected (LSG 2-5, LSG 3-7, and LSG 3-8) and defined by morphological and 16S rRNA analysis as Bacillus velezensis, Bacillus aryabhattai, and Bacillus mojavensis, respectively. Results showed the strain tolerant abilities to gastrointestinal fluid, bile salt, pH, and temperature expotures. Additionally, all bacterial strains showed anti-pathogenic activity against at least four strains out of six tested pathogen strains (Staphylococcus aureus, Aeromonas hydrophila, Escherichia coli, Aeromonas veronii, Edwardsiella, and Aeromonas sobria). The bacterial strains also showed a high percentage of co-aggregation activity, more than 70%, with Aer. hydrophile, Staph. epidermidis, and Klebsiella aerogenes. At the same time, the results of competition, rejection, and substitution activity with Aer. hydrophila and Aer. veronii indicated the ability of the isolated strains to reduce the adhesion of pathogens to mucin. All strains showed safety properties, non-hemolytic, and sensitivity characteristics for most of tested antibiotics. In vivo test after injecting these strains into fish at various concentrations showed no side effects in the internal or external organs of fish compared to controls, proving that this is safe for these fish. Furthermore, the three strains produced lipase, amylase, and protease enzymes. The strains also showed bile salt hydrolase activity and biofilm formation, allowing them to tolerate stressful conditions. Conclusion: Based on these strains characteristics and features, they could be considered a promising candidate probiotic and can be used as an anti-pathogenic, especially in aquaculture.

Exploration of the correlation between superficial cerebral veins identified using susceptibility-weighted imaging findings and cognitive differences between sexes based on deep learning: a preliminary study.

Background: This study aimed to investigate the association of superficial cerebral veins (SCVs) with sex-related cognitive differences and the possible hemodynamic mechanisms underlying these associations. Methods: This investigation was a prospective case-control study. A total of 344 healthy volunteers were recruited. In all, 200 volunteers were included to establish the deep learning model, and 144 volunteers were used for the research, including 72 males (50%) and 72 females (50%). No significant differences in age (P=0.358) or education (P=0.779) were observed between the sexes. Cognitive functioning was evaluated using neuropsychological tests, including the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment-Basic (MOCA-B). Susceptibility-weighted imaging scans were acquired with a 3.0 T magnetic resonance imaging system using a 32-channel high-resolution phased array coil. Minimum intensity projection images were obtained by reconstructing susceptibility-weighted imaging data. A deep learning model was trained on the minimum intensity projection images to quantify the diameter, tortuosity index, length, and the number of SCVs in the bilateral cerebral hemispheres. Finally, the association between cognitive differences between males and females and the properties of the SCVs was analyzed. Results: The MMSE and MOCA-B scores of males were significantly higher than those of females (P<0.05). Males had more SCVs in the bilateral cerebral hemispheres than did females (right hemisphere: P<0.01; left hemisphere: P<0.05). The number of SCVs in the right cerebral hemisphere was significantly and positively correlated with the MMSE and MOCA-B scores (correlation coefficients: 0.246 and 0.201, respectively; P<0.05). The number of SCVs in the left cerebral hemisphere was positively correlated with the MMSE scores (correlation coefficient: 0.196; P<0.05) and the MOCA-B scores. In this study, no significant correlations were observed between cognition and the diameter, length, or tortuosity index of the SCVs in the bilateral cerebral hemispheres. Conclusions: The cognitive function of males was better than that of females, and the different numbers of SCVs may be one of the explanations for this phenomenon of sex-based differences in cognition.

Effect of Bacillus amyloliquefaciens LSG2-8 on the intestinal barrier function of Amur minnow (Rhynchocypris lagowskii).

Probiotics are widely used in aquaculture. This article aims to study the effect of Bacillus amyloliquefaciens LSG2-8 on the intestinal barrier function of Rhynchocypris lagowskii. B. amyloliquefaciens LSG2-8 were added to R. lagowskii basal diets (CK) as additives at four concentrations: 1.0 × 106 (D-6), 1.0 × 107 (D-7), 1.0 × 108 (D-8) and 1.0 × 109 (D-9) CFU g-1 by dry weight of basal diet. After a 56-day feeding experiment, the activities of intestinal digestive enzymes and immunity-related enzymes of R. lagowskii on group D-6, D-7, D-8 and D-9 diet were significantly higher than the control (P < 0.05). In molecular experiments, the authors found that the levels of TGF-ß mRNA, IL-10 mRNA, ZO-1 mRNA and claudin-3 mRNA in group D-8 R. lagowskii were significantly higher (P < 0.05) than those of the control and other groups. Furthermore, the levels of IL-1ß and IL-8 mRNA of R. lagowskii on group D-6, D-7, D-8 and D-9 diet were significantly lower than those of the control (P < 0.05). In addition, the authors found that B. amyloliquefaciens LSG2-8 can regulate the intestinal flora balance and improve the intestinal structure of R. lagowskii. In conclusion, B. amyloliquefaciens LSG2-8 can improve the intestinal barrier function of R. lagowskii and can be used as a feed additive in aquaculture.

The molecular mechanisms of alpha-lipoic acid on ameliorating aflatoxin B1-induced liver toxicity and physiological dysfunction in northern snakehead (Channa argus).

This research aimed to evaluate the protective mechanism of alpha-lipoic acid (α-LA) on the food-borne aflatoxin B1 (AFB1) exposure-induced liver toxicity and physiological dysfunction in the northern snakehead (Channa argus). 480 fish (9.24±0.01 g) were randomly assigned to four treatment groups and fed with four experimental diets for 56 d including the control group (CON), AFB1 group (200 ppb AFB1), 600 α-LA group (600 ppm α-LA+200 ppb AFB1), and 900 α-LA group (900 ppm α-LA+200 ppb AFB1). The results revealed that 600 and 900 ppm α-LA attenuated AFB1-induced growth inhibition and immunosuppression in northern snakehead. 600 ppm α-LA significantly decreased the serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and lactate dehydrogenase levels, and AFB1 bioaccumulation, and attenuated the changes of hepatic histopathological and ultrastructure induced by AFB1. Moreover, 600 and 900 ppm α-LA significantly up-regulated phase I metabolism genes (cytochrome P450-1a, 1b, and 3a) mRNA expression, inhibited the levels of malondialdehyde, 8­hydroxy-2 deoxyguanosine and reactive oxygen species in the liver. Notably, 600 ppm α-LA significantly up-regulated the expression levels of nuclear factor E2 related factor 2 and its related downstream antioxidant molecules (heme oxygenase 1 and NAD(P)H: quinone oxidoreductase 1, etc.), increased the phase II detoxification enzyme-related molecules (glutathione-S-transferase and glutathione), antioxidant parameters (catalase and superoxide dismutase, etc.), and the expressions of Nrf2 and Ho-1 protein in the presence of AFB1 exposure. Furthermore, 600 and 900 ppm α-LA significantly reduced the characteristic indices of AFB1-induced endoplasmic reticulum stress (glucose-regulated protein 78 and inositol requiring enzyme 1, etc.), apoptosis (caspase-3 and cytochrome c, etc.) and inflammation (nuclear factor kappa B and tumor necrosis factor α, etc.), while increased the B-cell lymphoma-2 and inhibitor of κBα in the liver after being exposed to AFB1. To summarize, the above results indicate that dietary α-LA could modulate the Nrf2 signaling pathway to ameliorate AFB1-induced growth inhibition, liver toxicity, and physiological dysfunction in northern snakehead. Although the concentration of α-LA increased to 900 ppm from 600 ppm, the protective effects of the 900 ppm α-LA do not show an advantage over the 600 ppm α-LA, and even show inferiority in some respects. So that the recommended concentration of α-LA is 600 ppm. The present study provides the theoretical foundation for developing α-LA as the prevention and treatment of AFB1-induced liver toxicity in aquatic animals.

Replacing starch with resistant starch (Laminaria japonica) improves water quality, nitrogen and phosphorus budget and microbial community in hybrid snakehead (Channa maculata ♀ × Channa argus ♂).

It is essential to increase the use of carbohydrates as an energy source and improve protein synthesis and utilization to reduce ammonia nitrogen emissions. A 60-day cultural experiment was conducted to assess the impact of resistant starch (kelp meal, Laminaria japonica) replacing starch on water quality, nitrogen and phosphorus budget and microbial community of hybrid snakehead. Approximately 1350 experimental fish (11.4 ± 0.15 g) were randomly divided into control group (C, 20% starch) and four resistant starch groups: low replacement group (LR, 15% starch), medium replacement group (MR, 10% starch), high replacement group (HR, 5% starch) and full replacement group (FR, 0% starch). The crude protein and crude fat content of hybrid snakehead fish fed with the FR diet had the most significant improvement (P < 0.05). However, resistant starch also increased the effectiveness of nitrogen and phosphorus utilization in hybrid snakeheads, which decreased the proportion of total nitrogen and total phosphorus in tail water. The minimum nitrogen and phosphorus emission rate was when the starch level was 6.1%. Denitrifying microbes including Gemmobacter, Rhodobacter, Emticicia and Bosea have become much more prevalent in group FR (P < 0.05). In general, replacing starch with resistant starch can enhance the rate at which nitrogen and phosphorus are used in feeding, lessening water pollution and altering environmental microbial composition. PRACTITIONER POINTS: Resistant starch (RS) improves whole fish nutritional content. Resistant starch improves dietary nitrogen and phosphorus utilization. Resistant starch acts as a carbon source and encourages the colonization of denitrifying bacteria in water.

Insights into the Effects and Mechanism of Andrographolide-Mediated Recovery of Susceptibility of Methicillin-Resistant Staphylococcus aureus to ß-Lactam Antibiotics.

The frequent resistance associated with ß-lactam antibiotics and the high frequency of mutations in ß-lactamases constitute a major clinical challenge that can no longer be ignored. Andrographolide (AP), a natural active compound, has been shown to restore susceptibility to ß-lactam antibiotics. Fluorescence quenching and molecular simulation showed that AP quenched the intrinsic fluorescence of ß-lactamase BlaZ and stably bound to the residues in the catalytic cavity of BlaZ. Of note, AP was found to reduce the stability of the cell wall (CW) in methicillin-resistant Staphylococcus aureus (MRSA), and in combination with penicillin G (PEN), it significantly induced CW roughness and dispersion and even caused its disintegration, while the same concentration of PEN did not. In addition, transcriptome sequencing revealed that AP induced a significant stress response and increased peptidoglycan (PG) synthesis but disrupted its cross-linking, and it repressed the expression of critical genes such as mecA, blaZ, and sarA. We also validated these findings by quantitative reverse transcription-PCR (qRT-PCR). Association analysis using the GEO database showed that the alterations caused by AP were similar to those caused by mutations in the sarA gene. In summary, AP was able to restore the susceptibility of MRSA to ß-lactam antibiotics, mainly by inhibiting the ß-lactamase BlaZ, by downregulating the expression of critical resistance genes such as mecA and blaZ, and by disrupting CW homeostasis. In addition, restoration of susceptibility to antibiotics could be achieved by inhibiting the global regulator SarA, providing an effective solution to alleviate the problem of bacterial resistance. IMPORTANCE Increasingly, alternatives to antibiotics are being used to mitigate the rapid onset and development of bacterial resistance, and the combination of natural compounds with traditional antibiotics has become an effective therapeutic strategy. Therefore, we attempted to discover more mechanisms to restore susceptibility and effective dosing strategies. Andrographolide (AP), as a natural active ingredient, can mediate recovery of susceptibility of MRSA to ß-lactam antibiotics. AP bound stably to the ß-lactamase BlaZ and impaired its hydrolytic activity. Notably, AP was able to downregulate the expression of critical resistance genes such as mecA, blaZ, and sarA. Meanwhile, it disrupted the CW cross-linking and homeostasis, while the same concentration of penicillin could not. The multiple inhibitory effect of AP resensitizes intrinsically resistant bacteria to ß-lactam antibiotics, effectively prolonging the use cycle of these antibiotics and providing an effective solution to reduce the dosage of antibiotics and providing a theoretical reference for the prevention and control of MRSA.

Research progress on the association between mastitis and gastrointestinal microbes in dairy cows and the effect of probiotics.

Mastitis in dairy cows affects milk quality and thereby constrains the development of the dairy industry. A clear understanding of the pathogenesis of mastitis can help its treatment. Mastitis is caused by the invasion of pathogenic bacteria into the mammary gland through the mammary ducts. However, recent studies suggested that an endogenous entero-mammary pathway in dairy cattle might also be playing an important role in regulating mastitis. Also, probiotic intervention regulating host gut microbes has become an interesting tool to control mastitis. This review discusses the association of gastrointestinal microbes with mastitis and the mechanism of action of probiotics in dairy cows to provide new ideas for the management of mastitis in large-scale dairy farms.