Higher RPS Research Articles

Aeromonas hydrophila Bivalen Vaccines to Improve Body Resistance catfish (Pangasius hypophthalmus) Against Attacks Motile Aeromonas Septicemia

Motile aeromonas septicemia/MAS diseases due to Aeromonas hydrophila infection is an obstacle in cultivation of catfish (Pangasius hypophthalmus). MAS control can be done by administering vaccines. The purpose of this research is to get the right and effective dose of bivalent vaccine for controlling MAS at catfish. Both isolates will be used as vaccines are local strains of A. hydrophila CASO.01.G and SBMI.2. Both of them were isolated from disease catfish in the aquaculture ponds in Cindai Alus and Sungai Batang villages, Banjar Regency, South Kalimantan. Vaccines are made by heating (heat killed) at 100 o C for 60 minutes. A total of 13 test catfish measuring 9-13 cm were included in each aquarium. The design used was a completely randomized design with 5 treatments and 3 replications, namely bivalent vaccine with a dose of 10 9 cells/ml, 10 8 cells/ml, 10 7 cells/ml, 10 6 cells/ml, and control (PBS solution pH 7.0). The highest antibody titers in treatments vaccinated fish at a dose of 10 7 cells / ml (1,706.67). The effectiveness of the vaccine can be seen by the high RPS value of the treatment dose 10 9 cells/ml (97.22%); 10 8 cells/ml (100%); 10 7 cells/ml (97.22%); and 10 6 cells/ml (100%). The control MTD value was 1 day with 97.43% mortality, while in the vaccine treatment with dose of 10 7 cells/ml, there was a MTD repeat value for 1 day, but only 1 fish died. Water quality of this research are supportive and not a cause of fish death during the challenge test

Characterization of novel antigenic vaccine candidates for nile tilapia (Oreochromis niloticus) against Streptococcus agalactiae infection

Streptococcus agalactiae is one of the important pathogens responsible for high mortality and economic losses of the tilapia industry worldwide. Based on ten serovars of S. agalactiae infection, subunit vaccine with conserved antigens is promising strategy corresponding stimulated long-term immunity and provides protection for animals against different serotypes of S. agalactiae. In the present study, eight proteins (AP, AL, LivK, ESAT6, essA, essB, essC and esaA) were selected from the S. agalactiae serotype Ia genome as immunogenic antigens with bioinformation and immune experiment assays. These recombinant proteins were successfully obtained through expression in Escherichia coli and the immunogenicity was assessed in tilapia challenge model. The results showed that the recombinant proteins caused high-level-specific antibodies production and high lysozyme activities, suggesting that the recombinant proteins induced specific humoral immune response and innate immune response of tilapia. The signficant increase were observed in the cytokines levels of TNF-α, IL-1β, IFN-γ, cc1, cc2 and immune-related genes levels of CD8α and MHC factors in the spleen and head kidney tissues, suggesting that the recombinant proteins induced immune response of tilapia through cytokines signal pathway and activated high cytotoxic T-lymphocyte (CTL) activity of tilapia. Furthermore, vaccinated tilapia conferred high levels of protection against challenge with a lethal dose of highly virulent serovar Ⅰa (highest RPS was 91.60% in AL and essC protein groups). Our results indicated that the eight recombinant proteins induced high level of immune responses and offered protection against S. agalactiae infection, could be potential subunit vaccine candidates.

Effects of dietary Lactobacillus plantarum C20015 on growth, immunity, and disease resistance in koi carp

Koi carp (Cyprinus carpio) are well-known and highly valuable fish in the international ornamental fish community that have been plagued by Aeromonas veronii infections in recent years. Some A. veronii infections are serious enough to cause death. Lactobacillus plantarum, a probiotic, is one of the most environmentally friendly and safe alternatives to antibiotics for use as a feed additive for aquatic animals such as fish. The present study investigated the effects of L. plantarum C20015 on growth performance, immune responses, and eradication of A. veronii infection in koi carp. During a 6-week feeding experiment in an indoor aquarium and a 14-day A. veronii artificial infection experiment, a control diet group was fed with untreated feed twice a day, while experimental groups were given different concentrations of L. plantarum in their feed. The results indicated that the specific growth rate and percent weight gain of fish were significantly higher in the experimental groups than in the control diet group (P < 0.05). The activity of superoxide dismutase (SOD), peroxidase (POD), and lysozyme (LZM) and the levels of IgM antibody were significantly higher in the L. plantarum–supplemented groups than in the control diet group, and the enzymatic activity in the 108 CFU/g C20015 group was higher than that in the 107 CFU/g C20015 and 109 CFU/g C20015 groups (P < 0.01). In contrast, alkaline phosphatase (AKP) activity in the experimental groups was the highest at 42 days. All fish in the control diet group died 7 days after A. veronii infection. After 14 days, the relative percent survival (%, RPS) of the fish in the L. plantarum–supplemented groups was significantly higher than that of the fish in the control group, and the 108 CFU/g C20015 group had the highest RPS. Therefore, we first reported that fish fed with 108 CFU/g of L. plantarum C20015 could promote food intake and growth for koi carp.

A CpG-riched plasmid as vaccine adjuvant reduce antigen dose of an inactivated Vibrio anguillarum vaccine in turbot (Scophthalmus maximus L.)

Inactivated vaccines are often applied with adjuvants in commercial fish farming. Although some mineral or non-mineral oil adjuvants show efficient improvement with inactivated vaccines, but sometimes bring side effects such as tissue adhesion and granulomatous lesion at the injection site. CpG ODN is a novel type of soluble adjuvant which has been proved to possess excellent advantages in fish vaccine development. In this study, we designed a tandem sequence of CpG ODN synthesized in plasmid pcDNA 3.1, and an inactivated Vibrio anguillarum vaccine developed in our previous work was chosen for determining the efficiency of the CpG-riched plasmids (pCpG) as an adjuvant. Results showed that pCpG we designed can offer higher immunoprotection with the vaccine. Interestingly, even below the minimum immune dosage of the vaccine, a high RPS of 84% was observed once the vaccine was administrated with the pCpG. Serum specific antibody titer, superoxide dismutase and total protein were enhanced and some immune genes related to both innate and adaptive immune response were upregulated, implying an effective auxiliary function of the pCpG. Totally, our study suggested that the pCpG is a potential and available adjuvant for turbot vaccine development.

Urinary extracellular vesicle-associated MCP-1 and NGAL derived from specific nephron segments differ between calcium oxalate stone formers and controls.

Randall's plaque (RP; subepithelial calcification) appears to be an important precursor of kidney stone disease. However, RP cannot be noninvasively detected. The present study investigated candidate biomarkers associated with extracellular vesicles (EVs) in the urine of calcium stone formers (CSFs) with low (<5% papillary surface area) and high (≥5% papillary surface area) percentages of RP and a group of nonstone formers. RPs were quantitated via videotaping and image processing in consecutive CSFs undergoing percutaneous surgery for stone removal. Urinary EVs derived from cells of different nephron segments of CSFs (n = 64) and nonstone formers (n = 40) were quantified in biobanked cell-free urine by standardized and validated digital flow cytometer using fluorophore-conjugated antibodies. Overall, the number of EVs carrying surface monocyte chemoattractant protein (MCP)-1 and neutrophil gelatinase-associated lipocalin (NGAL) were significantly lower in CSFs compared with nonstone former controls (P < 0.05) but did not differ statistically between CSFs with low and high RPs. The number of EVs associated with osteopontin did not differ between any groups. Thus, EVs carrying MCP-1 and NGAL may directly or indirectly contribute to stone pathogenesis as evidenced by the lower of these populations of EVs in stone formers compared with nonstone formers. Validation of EV-associated MCP-1 and NGAL as noninvasive biomarkers of kidney stone pathogenesis in larger populations is warranted.

Dietary inclusion of Orange peels derived pectin and Lactobacillus plantarum for Nile tilapia (Oreochromis niloticus) cultured under indoor biofloc systems

A 12-week feeding trial was carried out to investigate possible effects of dietary orange peels derived pectin (OPDP) and Lactobacillus plantarum CR1T5 (LP) singularly or combined on innate immune response, disease resistance, and growth performance of Nile tilapia fingerlings under indoor biofloc system. The fingerlings were fed the following diets: diet 1 (0 g kg−1 OPDP and 0 CFU g−1 L. plantarum), diet 2 (10 g kg−1 OPDP), diet 3 (108 CFU g−1 L. plantarum), and diet 4 (10 g kg−1 OPDP +108 CFU g−1 L. plantarum). At the end of feeding trial, skin mucus parameters, serum immune parameters, and growth performance were measured. Ten randomly selected fish were used in a challenge test with Streptococcus agalactiae. The results indicated that supplementations of OPDP + LP or/and significantly (P < .05) increased growth performance, skin mucus and serum immunity responses. The highest values were revealed in fish fed both OPDP and LP vs. individual applications. However, no significant (P > .05) differences were observed between fish fed OPDP and LP. The challenge test revealed that the relative percent survival (RSP) in diet 2, diet 3, and diet 4 was 43.33%, 50.0%, and 70.0%, respectively. Among the supplemented groups, fish fed 10 g kg−1 OPDP + LP showed significant (P < .05) higher RPS and resistance to S. agalactiae than the other groups. The present results suggested that the combination of OPDP and LP could be considered as potential feed-additives for aquaculture farmed fish under indoor biofloc system.

Effects of corncob derived xylooligosaccharide on innate immune response, disease resistance, and growth performance in Nile tilapia (Oreochromis niloticus) fingerlings

An-eight week experiment was conducted to test efficacy of corncob derived xylooligosaccharides (CDXOS) on mucosal and serum immune, disease resistance, and growth performance of Nile tilapia (Oreochromis niloticus). Three hundred and twenty tilapia fingerlings (20.72 ± 0.02 g) were fed the following diets; 0 (Diet 1- control), added 5 – (Diet 2), 10 – (Diet 3), and 20 g kg−1 CDXOS (Diet 4). After 4 and 8 weeks feeding were mucosal, serum immune responses, and growth performance measured. A challenge test (15 days) against Streptococcus agalactiae was conducted after 8 weeks post-feeding. A significant (P < 0.05) stimulation of skin mucus lysozyme and peroxidase activities, as well as serum immune parameters (serum lysozyme, serum peroxidase, alternative complement activities, phagocytosis index, and respiratory burst activity) was noticed by feeding the fish CDXOS supplemented diets compared to the control group. Highest (P < 0.05) innate immune parameters were observed by feeding the fish 10 g kg−1 CDXOS vs. the other treatments. With regard to the challenge test, relative percent survival (RSP) of Nile tilapia fingerlings fed Diet 2, Diet 3, and Diet 4 was 34.78%, 60.87%, and 30.43%, respectively. Among the supplemented groups, dietary inclusion of 10 g kg−1 CDXOS revealed significant (P < 0.05) higher RPS and resistance towards S. agalactiae than the other groups. Regarding growth performance, final weight, weight gain, specific growth rate, and feed conversion ratio were remarkably (P < 0.05) improved in the CDXOS groups; highest improvement was observed in the 10 g kg−1 CDXOS treatment. In conclusion, inclusion of 10 g kg−1 CDXOS derived from corncob improved growth performance and health status of Nile tilapia fingerlings.

EP-1634: IOERT in locally recurrent high grade RPS: a retrospective single center analysis of 83 cases

EP-1634: IOERT in locally recurrent high grade RPS: a retrospective single center analysis of 83 cases

A live attenuated Vibrio anguillarum vaccine induces efficient immunoprotection in Tiger puffer (Takifugu rubripes)

Tiger puffer (Takifugu rubripes) is becoming an economic promising aquaculture species in China. However, the development of Tiger puffer breeding industry is restricted by some serious aquatic disease such as vibriosis. An effective live attenuated vaccine MVAV6203 was developed in our previous studies by curing the virulence plasmid pEIB1 and unmarked inframe-deletion of the aroC gene from the virulent V. anguillarum. Here, we evaluated whether this live vaccine was suitable for Tiger puffer against disease caused by Vibrio genus. The live vaccine show virulence attenuation in both juvenile and adult fish vaccinated with either a single or high dose (50-fold single dose). In addition, administration of the live vaccine shew limited growth in fish and did not affect fish body weight significantly, with no adverse impact on growth between vaccinated and saline control fish. Furthermore, increased expression of cytokines involved in pro-inflammatory (IL-1β, TNFα and IL-6), cell-mediated immunity inducing (IL-12p35, IL-12p40 and IL-18), antiviral/intracellular pathogen killing (I-IFN-1, IFN-γ and Mx), peripheral T cell expansion and survival controlling (IL-2, IL-7 and IL-15) and antigen processing maker and anti-inflammatory (MHC I and IL-10) were elicited significantly after the vaccination. These cellular responses correlate with protection against virulent strain challenge and high RPS of 90.67% and 80.31% in juvenile and adult fish were obtained, respectively. These data indicated for the first time that the live attenuated V. anguillarum vaccine is suitably applied for the development of an effective and safe vaccine for prevention of vibriosis in Tiger puffer aquaculture industry.

A biocompatible surfactant, methyl ester sulphonate (MES), as a precipitating ligand for protein purification

A biocompatible surfactant, methyl ester sulphonate (MES), was evaluated in protein surfactant precipitation. The amount of lysozyme precipitated by MES was found to be a strong function of both the molar ratio of surfactant to protein (Rp), and pH. Precipitation increased proportionally with an increase in Rp up to an optimum of 16, where full precipitation was achieved, while with pH variations electrostatic interactions were found to be the main driver of precipitation. The precipitate was recoverable by solvent extraction (54–56% activity recovery by acetone, ethanol, 50% acetone/ethanol, and methanol), and counter-ionic surfactant, TOMAC (83.3% activity recovery at Rr=1.5). Importantly, the structural integrity of the lysozyme recovered, either through solvent extraction or counter-ionic surfactant, was maintained, and this was confirmed by CD spectra and deconvolution. Precipitation with MES was compared to a conventional surfactant, AOT; in contrast with AOT, MES did not resolubilise the precipitate or cause structural transformations at higher Rps. MES exhibited better selectivity than AOT since it had minimal hydrophobic interactions in the absence of electrostatic interactions between the surfactant’s anionic group, and the protein’s positively charged groups (pH>pI). Hence, along with its economic benefits, and environmental features, this work has highlighted the potential of using MES in surfactant precipitation.

Thermodynamic study on dynamic water and organic vapor sorption on amorphous valnemulin hydrochloride

The sorption of water and organic vapors on valnemulin hydrochloride was determined by dynamic vapor sorption at 25 °C. The adsorption-desorption behavior of water vapor and a series of organic vapors was investigated to probe the structural changes in valnemulin hydrochloride before and after sorption. The isothermal adsorption equilibrium data was evaluated using Guggenheim-Anderson-deBoer (GAB) and Brunauer-Emmett-Teller (BET) models. The BET model is applicable only at low relative pressures (0.1⩽RP⩽0.4) while the GAB model is applicable in the whole range of RPs (0.1⩽RP⩽0.9). The sorption kinetics at high RPs was determined by fitting the sorption data to the Avrami equation and the sorption content vs. time relationship could be predicted by the Avrami equation. Finally, the possible sorption mechanism of valnemulin hydrochloride was also discussed. Open image in new window

Prediction of hemorrhagic transformation in acute ischemic stroke using permeability surface of CT perfusion

Objective To investigate the value of permeability surface (PS) in predicting hemorrhagic transformation (HT) in acute ischernic stroke (AIS) using CT peffusion (CTP).Methods The study included 31 consecutive patients who presented symptoms suggestive of an AIS for 3-9 h. All patients underwent CT examination (noncontrast CT,CTP).HT was determined by follow-up CT images.According to presence of HT,the AIS was divided into HT group (PSHT,11 patients) and non-HT group (PSNo-HT,20 patients).PS,cerebral blood flow (CBF),cerebral blood volume (CBV) and mean transit time (MTT) on both sides of brains were measured.The relative PS(rPS),relative CBF (rCBF),relative CBV(rCBV) and relative MTT(rMTT) were obtained by calculating the ratio of the values of bilateral regions.The rPS between PSHT and PSNo-HT was compared with an exact Wilcoxon signed-rank test. The rCBF,rCBV,rMTT and the PS of the ischemic side between PSHr and PSNo-HT were compared with independent-sample t test.Meanwhile,Spearman rank correlation analysis was conducted to analyze the relationship between the CTP parameters and HT.Results The PS value of ischemic side was (1.61 ±0.77) ml · min - 1 · 100 g-1 for the PSHT group,and the value was (0.91 ± 0.49) ml · min - 1 · 100 g- 1 for the PSNo-HT group.For the PSHT group,rPS,rCBF,rCBV,rMTT were 2.76 ±0.78,0.32 ±0.18,0.66 ±0.31,2.67 ±0.71,and for the PSNo-HT group,rPS,rCBF,rCBV,rMTT were 1.35 ±0.19,0.50±0.21,0.91 ±0.28,2.62 ± 1.31.Compared with PSNo-HT,PSHT had higher rPS and PS value,and there were significant statistical differences (U =0.000,t =3.070,P ＜0.01).But rCBF and rCBV values were lower in the PSHT group compared to the PSNo-HT group,and there were significant statistical differences (trCsF =2.343,trCBV =2.210,P ＜ 0.05).There was no significant statistical difference in rMTT between the two groups(t =0.118,P ＞ 0.05).Significant positive correlations were detected between the rPS and PS with HT(r=0.496,0.821,P ＜0.01).Conclusions The value of rPS is helpful in predicting HT in AIS.And it can be used as a predictor in determining clinical personalized treatment and thus reduce the incidence of adverse events. Key words: Cerebrovascular accident; Capillary permeability ; Tomography,X-ray computed

Morphology, cell growth, and polysaccharide production of Nostoc flagelliforme in liquid suspension culture at different agitation rates

Noscocflagelliforme is a terrestrial macroscopic cyanobacterium with high economic value. Free-living cells that were separated from a natural colony of N. flagelliforme were cultivated in a 20-L photobioreactor for 16 days at five agitation rates with impeller tip speeds at 0.3, 0., 0.8, 1.0, and 1.5 m·s−1. With different impeller tip speeds there were significant differences in the cell growth and polysaccharide production, and different types of cell colonies appeared because of different shear forces caused by agitation. At harvest time, cell concentrations with tip speeds of 0.8 and 1.0 m·s−1 were clearly higher than those with the other three tip speeds, but dry cell weights with the tip speeds of 0.3, 0.5, 0.8, and 1.0 m·s−1 were almost the same. The highest RPS (polysaccharide that released into liquid medium) production was obtained with the tip speeds of 0.8 and 1.0 m·s−1, while the highest EPS (polysaccharide that formed capsule or slime layer) production was obtained with the tip speed of 0.5 m·s−1. The tip speed of 1.5 m·s−1 was harmful for both cell growth and polysaccharide production, indicating that an appropriate shear force was needed in the liquid suspension culture of N. flagelliforme.

A new model for coalescence efficiency of drops in stirred dispersions

A model for coalescence efficiency of two drops embedded in an eddy has been developed. Unlike the other models which consider only head-on collisions, the model considers the droplets to approach at an arbitrary angle. The drop pair is permitted to undergo rotation while they approach each other. For coalescence to occur, the drops are assumed to approach each other under a squeezing force acting over the life time of eddy but which can vary with time depending upon the angle of approach. The model accounts for the deformation of tip regions of the approaching drops and, describes the rupture of the intervening film, based on stability considerations while film drainage is continuing under the combined influence of the hydrodynamic and van der Waals forces. The coalescence efficiency is defined as the ratio of the range of angles resulting in coalescence to the total range of all possible approach angles. The model not only reconciles the contradictory predictions made by the earlier models based on similar framework but also brings out the important role of dispersed-phase viscosity. It further predicts that the dispersions involving pure phases can be stabilized at high rps values. Apart from explaining the hitherto unexplained experimental data of Konno et al. qualitatively, the model also offers an alternate explanation for the interesting observations of Shinnar.