Chitosan inhibits Penicillium expansum possibly by binding to DNA and triggering apoptosis.

Chitosan is a natural polysaccharide that is abundant, biocompatible and exhibits effective antifungal activity against various pathogenic fungi. However, the potential intracellular targets of chitosan in pathogenic fungi and the way of activity of chitosan are far from well known. The present work demonstrated that chitosan could inhibit Penicillium expansum, the principal causal agent of postharvest blue mold decay on apple fruits, by binding to DNA and triggering apoptosis. UV-visible spectroscopy, fluorescence spectroscopy and electrophoretic mobility assay proved the interaction between chitosan and DNA, while atomic force microscope (AFM) observation revealed the binding morphology of chitosan to DNA. Chitosan could inhibit in vitro DNA replication, and cell cycle analysis employing flow cytometry demonstrated that cell cycle was retarded by chitosan treatment. Furthermore, the reactive oxygen species (ROS) assay and membrane potential analysis showed that apoptosis was induced in P. expansum cells after exposure to chitosan. In conclusion, our results confirmed that chitosan interacts with DNA and induces apoptosis. These findings are expected to provide a feasible theoretical basis and practical direction for the promoting and implementing of chitosan in plant protection and further illuminate the possible antifungal mechanisms of chitosan against fungal pathogens.

Characterization of Bacillus velezensis E2 with abilities to degrade ochratoxin A and biocontrol against Aspergillus westerdijkiae fc-1.

Ochratoxin A (OTA), primarily produced by the fungi belonging to the species of Aspergillus and Penicillium, is one of the most common mycotoxins found in cereals and fruits. In addition to resulting in huge economic losses, OTA contamination also poses considerable threat to human and livestock health. Microbial degradation of mycotoxins has been considered with great potential in mycotoxins decontamination. In a previous study, Bacillus velezensis E2 was isolated by our laboratory and showed appreciable inhibitory effect on Aspergillus flavus growth and aflatoxin production in rice grains. In this study, B. velezensis E2 was investigated for its ability to remove OTA and biocontrol against the ochratoxigenic Aspergillus westerdijkiae fc-1. The results revealed that B. velezensis E2 has considerable inhibitory effect on A. westerdijkiae fc-1 both on PDA medium and pear fruits, with inhibitory rate of 51.7% and 73.9%, respectively. In addition, its ability to remove OTA was evaluated in liquid medium and the results showed that more than 96.1% of OTA with an initial concentration of 2.5 µg/mL could be removed by B. velezensis E2 in 48 h. Further experiments revealed that enzymatic transformation and alkaline hydrolysis might be the main mechanisms related to OTA degradation by B. velezensis E2, with ring open ochratoxin α (OP-OTα) as a possible degradation product. Our study indicated that the B. velezensis E2 strain could be a potential bacterial candidate in biodegradation of OTA and biocontrol against A. westerdijkiae fc-1.

Cellular uptake of chitosan and its role in antifungal action against Penicillium expansum.

Chitosan has wide-spectrum antimicrobial activity but knowledge of its antifungal mechanism is still incomplete. In this study, transcriptome of Penicillium expansum upon chitosan treatment was analyzed by RNA-Seq. KEGG enrichment analysis revealed that endocytosis as well as other physiological pathways was regulated by chitosan treatment. Clathrin adaptor protein mu-subunit (PeCAM) gene, which encodes a protein associated with clathrin-dependent endocytosis, was up-regulated after chitosan treatment. Deletion of PeCAM resulted in changes of conidial, hyphal and colonial morphology. Confocal microscopy images of the distribution of fluorescein isothiocyanate-labeled chitosan confirmed cellular internalization of chitosan. However, deletion of PeCAM almost completely blocked uptake of chitosan into fungal cells and ΔPeCAM mutant exhibited less sensitivity to chitosan compared with wild type, suggesting that chitosan uptake is mediated by clathrin-dependent endocytosis and internalized chitosan also plays an important role in its antifungal activity. Collectively, our results provide a new insight into the antifungal mechanism of chitosan.

Isolation and characterization of a Pseudomonas poae JSU-Y1 with patulin degradation ability and biocontrol potential against Penicillium expansum.

Patulin, one of the most common mycotoxins produced primarily by the Penicillium, Aspergillus and Byssochlamys species, is often associated with fruits and fruit-based products. Biodegradation by microbes is an effective method to remove or detoxify mycotoxins. In this study, a bacterial strain with patulin degradation capability was selectively isolated using oxindole, an analogue to patulin, as the sole carbon source, and identified as Pseudomonas poae JSU-Y1 by phylogenetic analysis on the basis of 16S rRNA sequence. This isolated bacterium could inhibit the growth of Penicillium expansum both on plate medium and apple fruit with inhibition ratio of 30.3% and 44.9%, respectively. Up to 87.7% of the initial patulin (2.5 µg/mL) was removed after incubation with Pseudomonas poae JSU-Y1 in liquid medium at 30 °C for 72 h. When challenged with apple juice, 79% of patulin could be degraded by this isolated strain. Additionally, ascladiol was tentatively identified as the patulin degradation intermediate by LC-MS analysis. Taken together, the experiment results indicated that the isolated Pseudomonas poae JSU-Y1 would be a promising bacterial resource to control patulin contamination and toxigenic fungal growth in agricultural products.

Quality assessment of instant green tea using portable NIR spectrometer.

Caffeine and catechin are two main components of instant green tea, and are essential components of tea quality. This paper mainly focuses on the feasibility of rapidly determining instant green tea components by using a portable near infrared (NIR) spectrometer. The two main components (caffeine and catechin) were studied. In addition, the instrument performance levels of portable and benchtop NIR spectrometers were studied and compared. Quantitative models developed using portable and benchtop spectrometers for measuring caffeine, total catechins, and four individual catechins were established and compared. After preprocessing using standard normal variate (SNV), the Rp values of the caffeine, total catechins, (-)-epigallocatechin, (-)-epigallocatechin 3-gallate, (-)-epicatechin, and (-)-epicatechin gallate in the partial least squares models for a portable NIR spectrometer were 0.974, 0.962, 0.669, 0.945, 0.942 and 0.905, respectively. For a benchtop NIR spectrometer, Rp values were 0.993, 0.958, 0.883, 0.955, 0.966 and 0.936, respectively. Passing-Bablok regression method results indicated no significant differences between the two instruments. A genetic algorithm (GA) and the successive projections algorithm (SPA) were used to screen the wavelength of the NIR spectrum and establish the model. The GA obtained more robust modeling results. This study concludes that the developed portable spectroscopy system combined with appropriate variable selection methods can be effectively used for rapid determination of caffeine, total catechins, and four individual catechins in instant green tea.

Qualitative discrimination of Chinese dianhong black tea grades based on a handheld spectroscopy system coupled with chemometrics.

The evaluation of Chinese dianhong black tea (CDBT) grades was an important indicator to ensure its quality. A handheld spectroscopy system combined with chemometrics was utilized to assess CDBT from eight grades. Both variables selection methods, namely genetic algorithm (GA) and successive projections algorithm (SPA), were employed to acquire the feature variables of each sample spectrum. A partial least-squares discriminant analysis (PLS-DA) and support vector machine (SVM) algorithms were applied for the establishment of the grading discrimination models based on near-infrared spectroscopy (NIRS). Comparisons of the portable and benchtop NIRS systems were implemented to obtain the optimal discriminant models. Experimental results showed that GA-SVM models by the handheld sensors yielded the best predictive performance with the correct discriminant rate (CDR) of 98.75% and 100% in the training set and prediction set, respectively. This study demonstrated that the handheld system combined with a suitable chemometric and feature information selection method could successfully be used for the rapid and efficient discrimination of CDBT rankings. It was promising to establish a specific economical portable NIRS sensor for in situ quality assurance of CDBT grades.

Cognitive spectroscopy for evaluating Chinese black tea grades (Camellia sinensis): near-infrared spectroscopy and evolutionary algorithms.

BACKGROUND: Grading represents an essential criterion for the quality assurance of black tea. The main objectives of the study were to develop a highly robust model for Chinese black tea of seven grades based on cognitive spectroscopy. RESULTS: Cognitive spectroscopy was proposed to combine near-infrared spectroscopy (NIRS) with machine learning and evolutionary algorithms, selected feature information from complex spectral data and show the best results without human intervention. The NIRS measuring system was used to obtain the spectra of Chinese black tea samples of seven grades. The spectra acquired were preprocessed by standard normal variate transformation (SNV), multiplicative scatter correction (MSC) and minimum/maximum normalization (MIN/MAX), and the optimal pretreating method was implemented using principal component analysis combined with linear discriminant analysis algorithm. Three feature selection evolutionary algorithms, which were a genetic algorithm (GA), simulated annealing (SA) and particle swarm optimization (PSO), were compared to search the best preprocessed characteristic wavelengths. Cognitive models of Chinese black tea ranks were constructed using extreme learning machine (ELM), K-nearest neighbor (KNN) and support vector machine (SVM) methods based on the selected characteristic variables. Experimental results revealed that the PSO-SVM model showed the best predictive performance with the correlation coefficients of prediction set (Rp ) of 0.9838, the root mean square error of prediction (RMSEP) of 0.0246, and the correct discriminant rate (CDR) of 98.70%. The extracted feature wavelengths were only occupying 0.18% of the origin. CONCLUSION: The overall results demonstrated that cognitive spectroscopy could be utilized as a rapid strategy to identify Chinese black tea grades. © 2020 Society of Chemical Industry.

[Epidemiological analysis on leprosy in China, 2009].

OBJECTIVE: To study the epidemiological characteristics of leprosy and to provide evidence for specific strategies development on leprosy control. METHODS: Descriptive analysis of data from the national leprosy surveillance system was carried out in 2009. RESULTS: A total number of 1597 new cases were detected in 2009 with a case detection rate of 0.120 per 100 000 population. The proportions of children under 15, multibacillary and disability grade 2 were 2.4%, 84.3% and 22.8% respectively. A total number of 148 relapse cases were reported in 2009, among which 69 cases were those relapsed after multi-drug therapy. The existing cases were 6603 by the end of 2009 with a prevalence rate of 0.049 per 10 000 populations, among which 3332 cases were on multi-drug therapy. CONCLUSION: The overall leprosy situation is considered to be at low endemic in China but the distribution is unequal, with pocket areas in Yunnan, Sichuan and Guizhou. In order to reach the target on leprosy elimination, which was considered as a public health and social problem, the input and efforts are needed for many years to come.