RESUMEN

Citrobacter freundii, a common pathogen of freshwater fish, causes significant commercial losses to the global fish farming industry. In the present study, a highly pathogenic C. freundii strain was isolated and identified from largemouth bass (Micropterus salmoides). The pathogenicity and antibiotic sensitivity of the C. freundii strain were evaluated, and the histopathology and host immune response of largemouth bass infected with C. freundii were investigated. The results showed that C. freundii was the pathogen causing disease outbreaks in largemouth bass, and the infected fish showed typical signs of acute hemorrhages and visceral enlargement. Antimicrobial susceptibility testing showed that the C. freundii strain was resistant to Kanamycin, Medimycin, Clindamycin, Penicillin, Oxacillin, Ampicillin, Cephalexin, Cefazolin, Cefradine and Vancomycin. Histopathological analysis showed different pathological changes in major tissues of diseased fish. In addition, humoral immune factors such as superoxide dismutase (SOD), catalase (CAT) and lysozyme (LZM) were used as serum indicators to evaluate the immune response of largemouth bass after infection. Quantitative real-time PCR (qRT-PCR) was performed to investigate the expression pattern of immune-related genes (CXCR1, IL-8, IRF7, IgM, CD40, IFN-γ, IL-1ß, Hep1, and Hep2) in liver, spleen, and head kidney tissues, which demonstrated a strong immune response induced by C. freundii infection in largemouth bass. The present study provides insights into the pathogenic mechanism of C. freundii and immune response in largemouth bass, promoting the prevention and treatment of diseases caused by C. freundii infection.

Asunto(s)

RESUMEN

The largemouth bass virus (LMBV) isolate of Santee-Cooper ranavirus showed evidence of widespread infection in adult fish, but disease presentation caused by different viral strains exhibited considerable difference. In this study, a highly pathogenic LMBV-like resembling Santee-Cooper ranavirus was isolated and identified from juvenile largemouth bass. The pathogenicity and dynamic distribution of LMBV-like strain, histopathological analysis and host immune response of juvenile largemouth bass infected with LMBV-like were investigated. The results show that LMBV-like was highly pathogenic to juvenile fish, and the infected fish showed typical signs of acute haemorrhages and visceral enlargement. LMBV-like positive cells were found in the liver, spleen, kidney, gills, and intestinal tissue, and the virus content in spleen was the highest. Histopathological analysis showed different pathological changes in major tissues of diseased fish, mostly manifested as infiltration of inflammatory cell and histiocyte necrosis. In addition, humoral immune factors such as superoxide dismutase (SOD), catalase (CAT) and acid phosphatase (ACP) were used as serum indicators to evaluate the immune response of juvenile fish after infection. Quantitative real-time PCR (qRT-PCR) was used to evaluate the expression patterns of immune-related genes (CD40, IFN-γ, IgM, IL-1ß, IL-8, IL-12a, Mxd3, TGF-ß, and TNFα) in liver, spleen, and head kidney tissues. The results showed that immunological activity of the juvenile largemouth bass was significantly enhanced by LMBV-like infection. This research comprehensively systematically revealed the pathogenic characteristics of LMBV-like separated from juvenile largemouth bass and properties of the host's immune response caused by the virus infection, which providing a basis for further exploring the interaction between the virus and the host, and prevention and treatment of disease caused by Santee-Cooper ranavirus.

Asunto(s)

RESUMEN

Evaluation of polymer aging is very important for the long-term performance of polymer materials, but it remains a challenge to correlate accelerated evaluations with the real-time procedures. Here we develop a novel in-situ aging evaluation system for rapid and sensitive aging evaluations of polymer materials within hours under multiple environmental conditions. It is carried out by in-situ detecting the generation rate of trace gaseous degradation products, e.g. CO2, of polymer materials in a specially designed reaction cell during aging under environmental conditions with various UV irradiation, temperature and humidity. The advantages of this system were demonstrated by applying to evaluate the photo-oxidation of polypropylene (PP)-CaCO3 composites, including stability evaluation, aging status analysis, aging kinetics measurements and study on effects of UV irradiation intensity and humidity. The CO2 generation rate of PP-CaCO3 composites measured in this system is well correlated to carbonyl indices during 120-day natural weathering. A linear relationship was observed between the generation rate of CO2 and the natural logarithm of the carbonyl index. The activation energy of the photo-oxidation of PP-CaCO3 composites was calculated based on generation rates of CO2 at different temperatures in the range of 30-80 °C. The increase of UV irradiation intensity and humidity both enhanced the generation rate of CO2 of PP composites, and the presence of CaCO3 fillers promoted the sensitivity of PP photo-oxidation to both of UV irradiation intensity and humidity. This study provides a new approach to rapid and highly sensitive evaluation of polymer composite aging under multiple environmental conditions.

RESUMEN

Ferritin is a protein related to the storage of iron and widely distributed in animals. It participates in many biological process, including antioxidation, cell activation, angiogenesis, regulation of iron metabolic balance and immune defense. In the present study, a novel ferritin gene was identified from red swamp crayfish Procambarus clarkii, with a cDNA sequence encoding a predicted 221 amino-acid residues. The ferritin protein contains a 19-residue signal peptide and 145-residue classic ferritin domain. The NJ phylogenetic analysis showed PcFer clustered with other crustacean peptides. The recombinant PcFer protein was produced and purified in E. coli, and the anti-rabbit polyclonal antibody was obtained. The rPcFer exhibited iron binding activity at a dose-dependent effect. The qPCR and western blot analysis revealed that PcFer was highly expressed in hemocytes, hepatopancreas, and gills. After challenged with WSSV and Aeromonas hydrophila, the mRNA and protein expression patterns of PcFer were significantly up-regulated in hemocytes and hepatopancreas. dsRNA interfering technique was utilized to silence the expression of PcFer gene. The WSSV copy number in PcFer silenced shrimp was much higher than that in the control group. The present study indicated that PcFer was involved in the immune defense against WSSV and Aeromonas hydrophila, and might inhibit WSSV replication in P. clarkii. These results will provide important data support for further study of the functional role of the ferritin gene.

Asunto(s)

RESUMEN

Giant freshwater prawn Macrobrachium rosenbergii is an economically important species. However, its growth retardant have brought serious economic losses in recent years. Antibiotics abuse is suggested as a reason for M. rosenbergii's growth retardant, while few studies focused on the toxic effect of antibiotics on M. rosenbergii. To investigate the effect of enrofloxacin, a widely used antibiotic, on juvenile M. rosenbergii, a 14 days exposure study was carried out within 0.2, 1 and 5 mg/L enrofloxacin and followed by 7 days decontamination. Results showed that during the test period, enrofloxacin had the largest accumulation in juvenile shrimp at day 3, and gradually decreased at day 7 and 14, and almost all the drugs are cleared after 3 days decontamination. Short-term exposure to low dose enrofloxacin can promote the growth of juveniles. High dose enrofloxacin inhibited the growth of juvenile shrimp, to gill and liver damage, and induced apoptosis of the hepatopancreatic cells. These adverse effects was possibly caused by enrofloxacin-induced oxidative stress. Moreover, we also found the damage caused by high concentrations of enrofloxacin was irreversible in the short term. Collectively, these data indicated that enrofloxacin did affect the juvenile shrimp growth and development, and high level enrofloxacin abuse may contributed to M. rosenbergii's growth retardant.

Asunto(s)

RESUMEN

This research aimed to study the effects of astaxanthin on energy budget and bioaccumulation of microcystin-leucine-arginine (microcystin-LR) in the crayfish Procambarus clarkii (Girard, 1852). The crayfish (21.13 ± 4.6 g) were cultured under microcystin-LR stress (0.025 mg/L) and were fed with fodders containing astaxanthin (0, 3, 6, 9, and 12 mg/g) for 8 weeks in glass tanks (350 mm × 450 mm × 150 mm). Accumulations of microcystin-LR were measured in different organs of P. clarkii. The results suggested that astaxanthin can significantly improve the survival rate and specific growth rate (SGR) of P. clarkii (p < 0.05). The dietary astaxanthin supplement seems to block the bioaccumulation of microcystin-LR in the hepatopancreas and ovaries of P. clarkii to some extent (p < 0.05). Astaxanthin content of 9⁻12 mg/g in fodder can be a practical and economic choice.

Asunto(s)

RESUMEN

The ability to simultaneously detect JAK2 V617F and MPL W515K/L mutations would substantially improve the early diagnosis of myeloproliferative neoplasms (MPNs) and decrease the risk of arterial thrombosis. The goal of this study is to achieve a point of care testing platform for simultaneous analysis of major genetic alterations in MPN. Here, we report a microfluidic platform including a glass capillary containing polypropylene matrix that extracts genomic DNA from a drop of whole blood, a microchip for simultaneous multi-gene mutation screening, and a handheld battery-powered heating device. The µmLchip system was successfully used for point-of-care identification of the JAK2 V617F and MPL W515K/L mutations. The µmLchip assays were then validated by mutation analysis with samples from 100 MPN patients who had previously been analyzed via unlabeled probe melting curve analysis or real-time PCR. The results from the µmLchip were in perfect agreement with those from the other methods, except for one discrepant result that was negative in the unlabeled probe melting curve analysis but positive in the µmLchip. After T-A cloning, sequences of cloned PCR products revealed JAK2 V617F mutation in the sample. The portable microfluidic platform may be very attractive in developing point-of-care diagnostics for MPL W515K/L and JAK2 V617F mutations.

Asunto(s)