Thermostable vacuum foam dried Newcastle disease vaccine: Process optimization and pilot-scale study.

Vacuum foam drying (VFD) has been shown to improve the thermostability and long-term shelf life of Newcastle Disease Virus (NDV). This study optimized the VFD process to improve the shelf life of NDV at laboratory-scale and then tested the optimized conditions at pilot-scale. The optimal NDV to T5 formulation ratio was determined to be 1:1 or 3:2. Using the 1:1 virus to formulation ratio, the optimal filling volumes were determined to be 13-17% of the vial capacity. The optimized VFD process conditions were determined to be at a shelf temperature of 25℃ with a minimum overall drying time of 44 h. The vaccine samples prepared using these optimized conditions at laboratory-scale exhibited virus titer losses of ≤ 1.0 log10 with residual moisture content (RMC) below 3%. Furthermore, these samples were transported for 97 days around China at ambient temperature without significant titer loss, thus demonstrating the thermostability of the NDV-VFD vaccine. Pilot-scale testing of the NDV-VFD vaccine at optimized conditions showed promising results for up-scaling the process as the RMC was below 3%. However, the virus titer loss was slightly above 1.0 log10 (approximately 1.1 log10). Therefore, the NDV-VFD process requires further optimization at pilot scale to obtain a titer loss of ≤ 1.0 log10. Results from this study provide important guidance for possible industrialization of NDV-VFD vaccine in the future. KEY POINTS: • The process optimization and scale-up test of thermostable NDV vaccine prepared through VFD is reported for the first time in this study. • The live attenuated NDV-VFD vaccine maintained thermostability for 97 days during long distance transportation in summer without cold chain conditions. • The optimized NDV-VFD vaccine preparations evaluated at pilot-scale maintained acceptable levels of infectivity after preservation at 37℃ for 90 days, which demonstrated the feasibility of the vaccine for industrialization.

Effect of ultra-violet light radiation on Scenedesmus vacuolatus growth kinetics, metabolic performance, and preliminary biodegradation study.

This study presents the effect of ultra-violet (UV) light radiation on the process kinetics, metabolic performance, and biodegradation capability of Scenedesmus vacuolatus. The impact of the UV radiation on S. vacuolatus morphology, chlorophyll, carotenoid, carbohydrates, proteins, lipid accumulation, growth rate, substrate affinity and substrate versatility were evaluated. Thereafter, a preliminary biodegradative potential of UV-exposed S. vacuolatus on spent coolant waste (SCW) was carried out based on dehydrogenase activity (DHA) and total petroleum hydrocarbon degradation (TPH). Pronounced structural changes were observed in S. vacuolatus exposed to UV radiation for 24 h compared to the 2, 4, 6, 12 and 48 h UV exposure. Exposure of S. vacuolatus to UV radiation improved cellular chlorophyll (chla = 1.89-fold, chlb = 2.02-fold), carotenoid (1.24-fold), carbohydrates (4.62-fold), proteins (1.44-fold) and lipid accumulations (1.40-fold). In addition, the 24 h UV exposed S. vacuolatus showed a significant increase in substrate affinity (1/Ks) (0.959), specific growth rate (µ) (0.024 h-1) and biomass accumulation (0.513 g/L) by 1.50, 2 and 1.9-fold respectively. Moreover, enhanced DHA (55%) and TPH (100%) degradation efficiency were observed in UV-exposed S. vacuolatus. These findings provided major insights into the use of UV radiation to enhance S. vacuolatus biodegradative performance towards sustainable green environment negating the use of expensive chemicals and other unfriendly environmental practices.

Anthracene detoxification by Laccases from indigenous fungal strains Trichoderma lixii FLU1 and Talaromyces pinophilus FLU12.

The persistence and ubiquity of polycyclic aromatic hydrocarbons (PAHs) in the environment necessitate effective remediation strategies. Hence, this study investigated the potential of purified Laccases, TlFLU1L and TpFLU12L, from two indigenous fungi Trichoderma lixii FLU1 (TlFLU1) and Talaromyces pinophilus FLU12 (TpFLU12), respectively for the oxidation and detoxification of anthracene. Anthracene was degraded with vmax values of 3.51 ± 0.06 mg/L/h and 3.44 ± 0.06 mg/L/h, and Km values of 173.2 ± 0.06 mg/L and 73.3 ± 0.07 mg/L by TlFLU1L and TpFLU12L, respectively. The addition of a mediator compound 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) to the reaction system significantly increased the degradation of anthracene, with up to a 2.9-fold increase in vmax value and up to threefold decrease in Km values of TlFLU1L and TpFLU12L. The GC-MS analysis of the metabolites suggests that anthracene degradation follows one new pathway unique to the ABTS system-hydroxylation and carboxylation of C-1 and C-2 position of anthracene to form 3-hydroxy-2-naphthoic acid, before undergoing dioxygenation and side chain removal to form chromone which was later converted into benzoic acid and CO2. This pathway contrasts with the common dioxygenation route observed in the free Laccase system, which is observed in the second degradation pathways. Furthermore, toxicity tests using V. parahaemolyticus and HT-22 cells, respectively, demonstrated the non-toxic nature of Laccase-ABTS-mediated metabolites. Intriguingly, analysis of the expression level of Alzheimer's related genes in HT-22 cells exposed to degradation products revealed no induction of neurotoxicity unlike untreated cells. These findings propose a paradigm shift for bioremediation by highlighting the Laccase-ABTS system as a promising green technology due to its efficiency with the discovery of a potentially less harmful degradation pathway, and the production of non-toxic metabolites.

Relative abundance and the fate of human rotavirus in wastewater during treatment processes: identification of potential infectious rotavirus in the final effluents and receiving aquatic milieu in Durban area, South Africa.

The occurrence and persistence of rotaviruses in raw and treated wastewater and their discharge into rivers represent a significant health risk for humans and animals, worldwide. In this study, samples were collected monthly from each of the four Durban wastewater treatment plants (DWWTPs) and receiving rivers for a period of 3 months. Rotavirus was quantified by real-time quantitative PCR (RT-qPCR), and viability was assessed using integrated cell culture (ICC)-qPCR. Rotavirus was detected consistently in 100% of influent wastewaters (mean concentration range, 4.36-4.46 log10 genome equivalent (GE) copies/L) and final effluent samples of three DWWTPs (range, 3.35-3.61 log10 GE copies/L). Overall, 94% (45/48) of the wastewater analyzed and 95% (20/21) of the associated river water samples were positive for rotavirus (range, 2.04-6.77 log10 GE copies/L). The activated sludge process with 0.10-0.43 log10 reduction values (LRV) only moderately reduced the viral loads. Similarly, one of the DWWTPs that operated the biofilter modality produced 0.20 LRV. Though the additional treatment with chlorine produced higher LRV (range, 0.31-0.53) than the corresponding activated sludge or biofilter process, the difference in viral removals was not significant (p > 0.05). The equivalent treatment efficiencies of the four DWWTPs varied from 19 to 43% decay in the population of rotavirus. Further, infectious rotavirus ranging from 66.67 to 100%, 50 to 100%, and 66.67 to 100% were detected in the post-activated sludge, final effluents, and river water samples, respectively. In conclusion, the findings of infectious rotavirus in both the final effluents and associated rivers represent an infection risk for humans or animals during contact. Thus, close monitoring for rotavirus and risk assessment studies under distinct exposure scenarios may further shed light on the health-related risks associated with water recovery and reuse in urban settings.

Effect of Crop Rotation on Fusarium Mycotoxins and Fusarium Species in Cereals in Sichuan Province (China).

The present study was performed to evaluate the effect of crop rotation on Fusarium mycotoxins and species in cereals in Sichuan Province. A total of 311 cereal samples were randomly collected and analyzed from 2018 to 2019 in Sichuan Province. The results of mycotoxin analysis showed that the major trichothecene mycotoxins in Sichuan Province were nivalenol (NIV) and deoxynivalenol (DON), and the mean concentration of total trichothecenes (including NIV, fusarenone X [4ANIV], DON, 3-acetyldeoxynivalenol [3ADON], and 15-acetyldeoxynivalenol [15ADON]) in wheat was significantly higher than that in maize and rice. The concentration of total trichothecenes in the succeeding crops was significantly higher than that in the previous crops. In addition, wheat grown after maize had reduced incidence and concentration of trichothecene mycotoxins compared with that grown after rice, and ratooning rice grown after rice had increased incidence and concentration of trichothecene mycotoxins. Our data indicated that Fusarium asiaticum with the NIV chemotype was predominant in wheat and rice samples, while the number of the NIV chemotypes of F. asiaticum and Fusarium meridionale and the 15ADON chemotype of Fusarium graminearum in maize were almost the same. Although the composition of Fusarium species was affected by crop rotations, there were no differences when comparing the same crop rotation except for the maize-wheat rotation. Moreover, the same species and chemotype of Fusarium strains originated from different crops in various rotations, but there were no significant differences in pathogenicity in wheat and rice. These results contribute to the knowledge of the effect of crop rotation on Fusarium mycotoxins and species affecting cereals in Sichuan Province, which may lead to improved strategies for control of Fusarium mycotoxins and fungal disease in China.

Hibiscus sabdariffa (Roselle) calyx: a systematic and meta-analytic review of memory-enhancing, anti-neuroinflammatory and antioxidative activities.

BACKGROUND: In this study, we summarized the preclinical investigations of the neuroprotective activities of Hibiscus sabdariffa (HSD) extract via its effect on memory function, neuroinflammation and oxidative damage in the central nervous system, which may help to guide future studies. METHODS: Preclinical studies that investigated the effect of HSD extract on memory impairment, neuroinflammation and oxidative stress-induced neuronal damage were searched systematically in PubMed, EBSCOhost (including MEDLINE, CINAHL, APA PsycInfo, etc.), Web of Science (WoS) and Scopus. Parameters and indexes included Morris water maze, passive avoidance test, acetylcholinesterase activity, interleukin 1 (IL-1), tumour necrosis factor-alpha (TNF-α), MAPK, malondialdehyde (MDA), glutathione (GSH), reactive oxygen species (ROS) and mitochondria membrane potential (MMP). RESULTS: A total of 285 documents were identified; however, only ten articles were included and used for meta-analysis. The meta-analytic outcome revealed that HSD did not show any significant effect on memory function, neuroinflammatory biomarkers (IL-1, MAPK) and oxidative stress (GSH, MDA, ROS and MMP) in neuronal cells and tissues. CONCLUSIONS: Individual study revealed that HSD showed improved memory function, attenuated neuroinflammation and prevented oxidative damage to neurons. However, a conflicting result was observed from the meta-analytic outcomes which showed that HSD has no significant effect on cognitive impairment, neuroinflammation and oxidative stress-induced neuronal damage. However the contradiction in this finding may be associated with small number of studies included. Hence, more studies on the memory-enhacing effects and anti-neuroinflammatory activity of HSD in preclinical and clinical model are required to validate its neuroprotective effect.

A multifunctional enolase mediates cytoadhesion and interaction with host plasminogen and fibronectin in Mycoplasma hyorhinis.

Mycoplasma hyorhinis may cause systemic inflammation of pigs, typically polyserositis and arthritis, and is also associated with several types of human cancer. However, the pathogenesis of M. hyorhinis colonizing and breaching the respiratory barrier to establish systemic infection is poorly understood. Glycolytic enzymes are important moonlighting proteins and virulence-related factors in various bacteria. In this study, we investigated the functions of a glycolytic critical enzyme, enolase in the infection and systemic spread of M. hyorhinis. Bacterial surface localization of enolase was confirmed by flow cytometry and colony hybridization assay. Recombinant M. hyorhinis enolase (rEno) was found to adhere to pig kidney (PK-15) cells, and anti-rEno serum significantly decreased adherence. The enzyme was also found to bind host plasminogen and fibronectin, and interactions were specific and strong, with dissociation constant (KD) values of 1.4 nM and 14.3 nM, respectively, from surface plasmon resonance analysis. Activation of rEno-bound plasminogen was confirmed by its ability to hydrolyze plasmin-specific substrates and to degrade a reconstituted extracellular matrix. To explore key sites during these interactions, C-terminal lysine residues of enolase were replaced with leucine, and the resulting single-site and double-site mutants show significantly reduced interaction with plasminogen in far-Western blotting and surface plasmon resonance tests. The binding affinities of all mutants to fibronectin were reduced as well. Collectively, these results imply that enolase moonlights as an important adhesin of M. hyorhinis, and interacts with plasminogen and fibronectin. The two lysine residues in the C-terminus are important binding sites for its multiple binding activities.

Vacuum Foam Drying Method Improved the Thermal Stability and Long-Term Shelf Life of a Live Attenuated Newcastle Disease Virus Vaccine.

Vaccines used for managing Newcastle disease virus (NDV) rely heavily on cold chain, and this results in major constraints in their successful application, shipping, and storage. This study was undertaken to improve the thermotolerance properties of live attenuated NDV vaccines using vacuum foam drying (VFD) technology. The live attenuated NDV vaccine formulated in 15% trehalose, 2.5% gelatin, 0.05% pluronic, and 25 mmol/L potassium phosphate buffer (T5) and dried using VFD showed improved heat tolerance in comparison to the vaccine formulated in T5 as well, but dried using freeze-drying (FD) method. The T5-formulated VFD vaccine was stored at 37°C for 120 days, 45°C for 7 days, and 60°C for 3 days; the virus titer loss decreased by no more than 1.0 Log10. In contrast, the FD vaccine prepared in T5 could only be stored at 37°C for 7-10 days. Furthermore, the T5-formulated NDV-VFD vaccine remained infectious when heated at 100°C for 30 min. Shelf-life studies confirmed the improved thermal tolerance of the T5-formulated NDV-VFD vaccine since it could be stably stored at 2-8°C for 42 months and 25°C for 15 months. Moreover, immunization of 1-month-old specific pathogen-free (SPF) chickens with the T5-formulated NDV-VFD vaccine stored at 25 and 37°C could produce hemagglutination inhibition (HI) antibody levels comparable to those of commercial NDV-FD vaccines, which require strict adherence to the cold chain. In conclusion, not only did the VFD technology improve the thermostability and long-term shelf life of the vaccine, it also maintained its immunogenicity.

Whole-genome sequencing, genome mining, metabolic reconstruction and evolution of pentachlorophenol and other xenobiotic degradation pathways in Bacillus tropicus strain AOA-CPS1.

A pentachlorophenol degrading bacterium was isolated from effluent of a wastewater treatment plant in Durban, South Africa, and identified as Bacillus tropicus strain AOA-CPS1 (BtAOA). The isolate degraded 29% of pentachlorophenol (PCP) within 9 days at an initial PCP concentration of 100 mg L-1 and 62% of PCP when the initial concentration was set at 350 mg L-1. The whole-genome of BtAOA was sequenced using Pacific Biosciences RS II sequencer with the Single Molecule, Real-Time (SMRT) Link (version 7.0.1.66975) and analysed using the HGAP4-de-novo assembly application. The contigs were annotated at NCBI, RASTtk and PROKKA prokaryotic genome annotation pipelines. The BtAOA genome is comprised of a 5,246,860-bp chromosome and a 58,449-bp plasmid with a GC content of 35.4%. The metabolic reconstruction for BtAOA showed that the organism has been naturally exposed to various chlorophenolic compounds including PCP and other xenobiotics. The chromosome encodes genes for core processes, stress response and PCP catabolic genes. Analogues of PCP catabolic gene (cpsBDCAE, and p450) sequences were identified from the NCBI annotation data, PCR-amplified from the whole genome of BtAOA, cloned into pET15b expression vector, overexpressed in E. coli BL21 (DE3) expression host, purified and characterized. Sequence mining and comparative analysis of the metabolic reconstruction of the BtAOA genome with closely related strains suggests that the operon encoding the first two enzymes in the PCP degradation pathway were acquired from a pre-existing pterin-carbinolamine dehydratase subsystem. The other two enzymes were recruited via horizontal gene transfer (HGT) from the pool of hypothetical proteins with no previous specific function, while the last enzyme was recruited from pre-existing enzymes from the TCA or serine-glyoxalase cycle via HGT events. This study provides a comprehensive understanding of the role of BtAOA in PCP degradation and its potential exploitation for bioremediation of other xenobiotic compounds.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) moonlights as an adhesin in Mycoplasma hyorhinis adhesion to epithelial cells as well as a plasminogen receptor mediating extracellular matrix degradation.

Mycoplasma hyorhinis infects pigs causing polyserositis and polyarthritis, and has also been reported in a variety of human tumor tissues. The occurrence of disease is often linked with the systemic invasion of the pathogen. Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), one of the key enzymes of glycolysis, was reported as a surface multifunctional molecule in several bacteria. Here, we investigated whether GAPDH could manifest binary functions; as an adhesin to promote colonization as well as a plasminogen receptor functioning in extracellular matrix (ECM) degradation to promote systemic invasion. The surface localization of GAPDH was observed in M. hyorhinis with flow cytometry and colony blot analysis. Recombinant GAPDH (rGAPDH) was found to be able to bind porcine-derived PK-15 and human-derived NCI-H292 cells. The incubation with anti-GAPDH antibody significantly decreased the adherence of M. hyorhinis to both cell lines. To investigate its function in recruiting plasminogen, firstly, the interaction between rGAPDH and plasminogen was demonstrated by ELISA and Far-Western blot assay. The activation of the rGAPDH-bound plasminogen into plasmin was proved by using a chromogenic substrate, and furtherly confirmed to degrade extracellular matrix by using a reconstituted ECM. Finally, the ability of rGAPDH to bind different ECM components was demonstrated, including fibronectin, laminin, collagen type IV and vitronectin. Collectively, our data imply GAPDH as an important adhesion factor of M. hyrohinis and a receptor for hijacking host plasminogen to degrade ECM. The multifunction of GAPDH to bind both plasminogen and ECM components is believed to increase the targeting of proteolysis and facilitate the dissemination of M. hyorhinis.

Analysis of the Fusarium graminearum Species Complex from Gramineous Weeds Near Wheat Fields in Jiangsu Province, China.

Several weed species are known as alternative hosts of the Fusarium graminearum species complex (FGSC), and their epidemiological significance in Fusarium head blight (FHB) has been investigated; however, scant information is available regarding FGSC occurrence in weeds near Chinese wheat fields. To evaluate the potential role of gramineous weeds surrounding wheat fields in FHB, 306 FGSC isolates were obtained from 210 gramineous weed samples in 2018 in Jiangsu Province. Among them, 289 were Fusarium asiaticum, and the remainder were F. graminearum. Trichothecene genotype and mycotoxin analyses revealed that 74.3% of the F. asiaticum isolates were the 3-acetyldeoxynivalenol (3ADON) chemotype, and the remainder were the nivalenol (NIV) chemotype. Additionally, 82.4% of F. graminearum isolates were the 15-acetyldeoxynivalenol (15ADON) chemotype, and the remainder were the NIV chemotype. FHB severity and trichothecene analysis indicated that F. asiaticum isolates with the 3ADON chemotype were more aggressive than those with the NIV chemotype in wheat. 3ADON and NIV chemotypes of F. asiaticum isolated from weeds and wheat showed no significant differences in pathogenicity in wheat. All selected F. asiaticum isolates produced perithecia, with little difference between the 3ADON and NIV chemotypes. These results highlight the epidemiology of the FGSC isolated from weeds near wheat fields, with implications for reducing FHB inoculum in China.

Chemical Characterization, Antiproliferative and Antioxidant Activities of Polyunsaturated Fatty Acid-Rich Extracts from Chlorella sp. S14.

Microalgae is a rich source of polyunsaturated fatty acid. This study was conducted to identify and isolate microalgal strain with the potentials for producing polyunsaturated fatty acids (PUFAs) and determine its cytotoxic effect on some cancer cells. The algal strain (Chlorella sp. S14) was cultivated using modified BG-11 media, and algal biomass obtained was used for fatty acid extraction. Gas chromatographic-mass spectrometry was used to identify and quantify the levels of the fatty acid constituents. The total content of monounsaturated fatty acids (1.12%) was low compared to polyunsaturated fatty acids (PUFAs) (52.87%). Furthermore, n-3 PUFAs accounted for (12.37%) of total PUFAs with the presence of α-linolenic acid (2.16%) and cis-11,14,17-eicosatrienoic acid (2.16%). The PUFA-rich extract did not exhibit a cytotoxic effect on normal cells. Treatment with the PUFA-rich extract (150 µg/mL) significantly reduced cell viability in MCF-7 (31.58%) and A549 (62.56%) cells after the 48 h treatment. Furthermore, treatment of MCF-7 with fatty acid extracts (125 and 150 µg/mL) showed a significant reduction in MDA levels, increase in catalase activities and decrease in GSH level compared to untreated cells. However, a slight decrease in MDA level was observed in A549 cells after the 48 h treatment. There are no significant changes in catalase activities and GSH level in treated A549 cells. However, a slight reduction of NO levels was observed in treated MCF-7 and A549 cells. These results indicate the potentials of PUFA-rich extracts from Chlorella sp. S14 to reduce viability and modulate redox status in A549 and MCF-7 cells.

Applications of Lactic Acid Bacteria and Their Bacteriocins against Food Spoilage Microorganisms and Foodborne Pathogens.

Lactic acid bacteria (LAB) are Gram-positive and catalase-negative microorganisms used to produce fermented foods. They appear morphologically as cocci or rods and they do not form spores. LAB used in food fermentation are from the Lactobacillus and Bifidobacterium genera and are useful in controlling spoilage and pathogenic microbes, due to the bacteriocins and acids that they produce. Consequently, LAB and their bacteriocins have emerged as viable alternatives to chemical food preservatives, curtesy of their qualified presumption of safety (QPS) status. There is growing interest regarding updated literature on the applications of LAB and their products in food safety, inhibition of the proliferation of food spoilage microbes and foodborne pathogens, and the mitigation of viral infections associated with food, as well as in the development of creative food packaging materials. Therefore, this review explores empirical studies, documenting applications and the extent to which LAB isolates and their bacteriocins have been used in the food industry against food spoilage microorganisms and foodborne pathogens including viruses; as well as to highlight the prospects of their numerous novel applications as components of hurdle technology to provide safe and quality food products.

Characterization of DNA methylation-based markers for human body fluid identification in forensics: a critical review.

Body fluid identification in crime scene investigations aids in reconstruction of crime scenes. Several studies have identified and reported differentially methylated sites (DMSs) and regions (DMRs) which differ between forensically relevant tissues (tDMRs) and body fluids. Diverse factors affect methylation patterns such as the environment, diets, lifestyle, disease, ethnicity, genetic variation, amongst others. Thus, it is important to analyse the stability of markers employed for forensic identification. Furthermore, even though epigenetic modifications are described as stable and heritable, epigenetic inheritance of potential markers for body fluid identification needs to be assessed in the long term. Here, we discuss the current status of reported DNA methylation-based markers and their verification studies. Such thorough investigation is crucial to develop a stable panel of DNA methylation-based markers for accurate body fluid identification.

Biotransformation of pentachlorophenol by an indigenous Bacillus cereus AOA-CPS1 isolated from wastewater effluent in Durban, South Africa.

Pentachlorophenol (PCP) is a recalcitrant biocide that bioaccumulates in the environment due to its persistent nature and has been listed as a priority pollutant due to its toxicological and health effects. In this study, a novel PCP-degrading Bacillus cereus strain AOA-CPS1 (BcAOA) was isolated from wastewater and characterized for PCP biotransformation in a batch reactor. The degradation kinetics were elucidated via substrate inhibition models, while PCP biotransformation was established by spectrophotometric and GC-MS analysis. BcAOA shared 95% sequence homology with Bacillus cereus strain XS2 and is closely related to some B. cereus strains which are previously reported to degrade PCP and other related pollutants. BcAOA degraded 74% of 350 mg l-1 of PCP within 9 days in a batch culture. The biotransformation of PCP by BcAOA followed the first and zero-order kinetics at low and high PCP concentration, respectively, with biokinetic constants: maximum biotransformation rate (0.0996 mg l-1 h-1); substrate inhibition constant (723.75 mg l-1); half-saturation constant (171.198 mg l-1) and R2 (0.98). The genes (pcpABCDE, cytochrome P450) encoding the enzymes involved in the biodegradation of PCP were amplified from the genomic DNA of BcAOA. Further, depending upon the genes amplified and identified metabolites using GC-MS, two different PCP biotransformation pathways were proposed in this study. Cloning and expression of the catabolic genes are underway to map out the concise pathway for PCP biotransformation by BcAOA.

Analysis of Fusarium graminearum Species Complex from Freshly Harvested Rice in Jiangsu Province (China).

Members of Fusarium graminearum species complex (FGSC) are the major pathogens that cause Fusarium head blight (FHB) in cereals worldwide. Symptoms of FHB on rice, including dark staining or browning of rice glumes, were recently observed in Jiangsu Province, China. To improve our understanding of the pathogens involved, 201 FGSC isolates were obtained from freshly harvested rice samples and identified by phylogenetic analyses. Among the 201 FGSC isolates, 196 were F. asiaticum and the remaining 5 were F. graminearum. Trichothecene chemotype and chemical analyses showed that 68.4% of the F. asiaticum isolates were the 3-acetyldeoxynivalenol (3ADON) chemotype and the remainder were the nivalenol (NIV) chemotype. All of the F. graminearum isolates were the 15-acetyldeoxynivalenol chemotype. Pathogenicity assays showed that both the 3ADON and NIV chemotypes of F. asiaticum could infect wheat and rice spikes. FHB severity and trichothecene toxin analysis revealed that F. asiaticum with the NIV chemotype was less aggressive than that with the 3ADON chemotype in wheat, while the NIV-producing strains were more virulent than the 3ADON-producing strains in rice. F. asiaticum isolates with different chemotypes did not show significant differences in mycelial growth, sporulation, conidial dimensions, or perithecial production. These findings would provide useful information for developing management strategies for the control of FHB in China.

Neuroprotective effects of some seaweeds against Zn - induced neuronal damage in HT-22 cells via modulation of redox imbalance, inhibition of apoptosis and acetylcholinesterase activity.

Zinc plays an important role in neuronal signaling and neurotransmission. However, dyshomeostasis of this metal or its accumulation in the brain has been linked with neurological disorders such as Alzheimer's disease and Parkinson's disease. In this study, the neuroprotective effects of Ecklonia maxima (KPM), Gracilaria gracilis (GCL), Ulva lactuca (ULT) and Gelidium pristoides (MNP) in Zn -induced neurotoxicity in HT-22 cells was examined. Cells were treated with Zinc sulphate and/or aqueous - ethanol extracts and cell viability, apoptosis, acetylcholinesterase activity, including some antioxidant enzymes (catalase and superoxide dismutase activity) and glutathione (GSH) levels were determined. Malondialdehyde and nitric oxide levels produced in the Zn and/or seaweed extract treated cells were also determined. Prior treatment with the seaweed extracts improved cell viability and inhibited Zn - induced cell death. Acetylcholinesterase activity was significantly high in Zn treated cells compared to the control. Pre-treatment with the seaweed extracts triggered a decrease in acetylcholinesterase activity in Zn - treated cells. Furthermore, treatment with Zn caused a significant reduction in GSH levels as well as a decrease in superoxide dismutase and catalase activities. In contrast, the seaweed extract increased antioxidant enzyme activities and GSH levels. An increase in malondialdehyde and nitric oxide levels was also reversed after treatment with the seaweed extracts. These results suggest that the seaweed extracts improved cholinergic transmission disrupted by Zn - induced neurotoxicity and protected the cells against oxidative damage and neuroinflammation. The neuroprotective effects of the seaweed extracts could be linked to their bioactive constituents. Hence these seaweeds are potential sources of active ingredients with neuroprotective potentials and could be used for the development of functional foods and/or nutraceuticals.

Macroalgae as a Valuable Source of Naturally Occurring Bioactive Compounds for the Treatment of Alzheimer's Disease.

Alzheimer's disease (AD) is a neurological condition that affects mostly aged individuals. Evidence suggests that pathological mechanisms involved in the development of AD are associated with cholinergic deficit, glutamate excitotoxicity, beta-amyloid aggregation, tau phosphorylation, neuro-inflammation, and oxidative damage to neurons. Currently there is no cure for AD; however, synthetic therapies have been developed to effectively manage some of the symptoms at the early stage of the disease. Natural products from plants and marine organisms have been identified as important sources of bioactive compounds with neuroprotective potentials and less adverse effects compared to synthetic agents. Seaweeds contain several kinds of secondary metabolites such as phlorotannins, carotenoids, sterols, fucoidans, and poly unsaturated fatty acids. However, their neuroprotective effects and mechanisms of action have not been fully explored. This review discusses recent investigations and/or updates on interactions of bioactive compounds from seaweeds with biomarkers involved in the pathogenesis of AD using reports in electronic databases such as Web of science, Scopus, PubMed, Science direct, Scifinder, Taylor and Francis, Wiley, Springer, and Google scholar between 2015 and 2019. Phlorotannins, fucoidans, sterols, and carotenoids showed strong neuroprotective potentials in different experimental models. However, there are no data from human studies and/or clinical trials.

Antibiogram and molecular characterization of methicillin-resistant Staphylococcus aureus recovered from treated wastewater effluent and receiving surface water in Durban, South Africa.

Municipal wastewater treatment plants (WWTPs) may serve as a reservoir for potentially pathogenic and antibiotic resistant bacteria. The discharge of improperly treated wastewater effluent may lead to the spread of these bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) which is responsible for causing pneumonia, septicaemia and skin and soft tissue infections, into the receiving surface waters. This study aimed to determine the antibiogram and virulence gene profiles of MRSA isolates recovered from treated wastewater effluent and receiving surface waters. Genetic fingerprinting of the isolates was also carried out to determine the phylogenetic relationship between the isolates and selected antibiogram profiles. Eighty MRSA isolates were obtained from treated effluent and receiving rivers of two WWTPs in Durban, KwaZulu-Natal. Antibiotic resistance was observed towards lincomycin (100%), oxacillin (98.75%), cefoxitin and penicillin (97.50%), and ampicillin (96.25%). In addition, 72.50%, 66.25%, 52.50%, 40% and 33.75% of isolates showed resistance against cefozolin, azithromycin, amoxicillin/clavulanic acid, erythromycin and vancomycin, respectively. Antibiotic resistance genes detected in the isolates tested in this study: aac(6')/aph(2″) (56.25%), ermC (62.50%), msrA (22.50%), and blaZ and tetK (70%). The frequency of virulence genes: hla and sea was 57.50%, hld was 1.25%, while lukS P/V was 0%. Pulse Field Gel Electrophoresis analysis generated 13 pulsotypes (designated A-M) showing a correlation to their respective antibiograms. Findings from this study showed the presence of potentially pathogenic, multi-drug resistant MRSA in the treated effluent and receiving surface waters. This may have detrimental effects on the health of individuals who come into contact with these water resources.

Phenolic composition, antioxidant activity, anticholinesterase potential and modulatory effects of aqueous extracts of some seaweeds on ß-amyloid aggregation and disaggregation.

Context: Seaweeds contain bioactive compounds with different biological activities. They are used as functional ingredients for the development of therapeutic agents to combat degenerative diseases. Objective: This study investigated the phenolic composition, antioxidant activity, cholinesterase inhibitory and anti-amyloidogenic activities of aqueous extracts of Gracilaria beckeri (J.Agardh) Papenfuss (Gracilariaceae) (RED-AQ), Ecklonia maxima (Osbeck) Papenfuss (Lessoniaceae) (ECK-AQ), Ulva rigida (C.Agardh) Linnaeus (Ulvaceae) (URL-AQ) and Gelidium pristoides (Turner) Kützing (Gelidiaceae) (GEL-AQ). Materials and methods: Phenolic composition of the seaweed extracts was determined using liquid chromatography mass spectrometry. Radical scavenging and metal chelating activities were assessed in vitro. The effect of the extracts (21-84 µg/mL) on acetylcholinesterase and butyrylcholinesterase activities were also investigated using an in vitro colorimetric assay. Transmission electron microscope and thioflavin-T fluorescence assay were used to examine the anti-amyloidogenic activities of the extracts. Results: Phloroglucinol, catechin, epicatechin 3-glucoside were identified in the extracts. ECK-AQ (IC50=30.42 and 280.47 µg/mL) exhibited the highest OH• scavenging and metal chelating activities, while RED-AQ (41.23 and 334.45 µg/mL) exhibited the lowest. Similarly, ECK-AQ (IC50 = 49.41 and 52.11 µg/mL) exhibited higher inhibitory effects on acetylcholinesterase and butyrylcholinesterase activities, while RED-AQ (64.56 and 63.03 µg/mL) showed the least activities. Rapid formation of ß-amyloid (Aß1-42) fibrils and aggregates was observed in electron micrographs of the control after 72 and 96 h. The reduction of Aß1-42 aggregates occurred after co-treatment with the seaweed extracts. Discussion and conclusion: ECK-AQ, GEL-AQ, URL-AQ and RED-AQ may possess neuroprotective potential and could be explored for the management of Alzheimer's disease.