Comparing the effects of three in situ methods on nitrogen loss control, temperature dynamics and maturity during composting of agricultural wastes with a stage of temperatures over 70 °C.

The study investigated the effects of three in situ methods for controlling nitrogen loss and maturity with different mechanisms: struvite-based addition (K2HPO4 and MgO, MP), woody peat addition (WP) and intermittent aeration (IA), during composting of vegetable waste (cucumber vine) with temperature over 70 °C to inactivate potential viral pathogens. The experiment was conducted in a 200 L pilot-scale composting system, with which temperature and ammonia emission were recorded in real time, and solid samples were collected and analyzed during the process. The results indicated that the methods of MP and IA reduced the total nitrogen loss by 27.5% and 16.1%, respectively, without inhibitory effects on the temperature, nutrient availability and maturity. The WP method significantly decreased the nitrogen loss but could not maintain the thermophilic stage over 70 °C, because of its influence on the material physio-chemical characteristics caused by woody peat addition. In conclusion, all three methods could promote the maturity process, and 20 days should be adequate for vegetable waste composting with a good nutrient availability. Considering the two factors of reducing nitrogen loss and achieving high temperatures together, we recommended the struvite-based controlling method with the mechanism of chemisorption to reduce nitrogen loss during vegetable waste composting that requires temperatures over 70 °C.

Stable isotope labelling by amino acids in cell culture (SILAC) applied to quantitative proteomics of Edwardsiella tarda ATCC 15947 under prolonged cold stress.

Edwardsiella tarda poses a threat to human health and has resulted in enormous economic losses in aquaculture. Low temperatures are usually applied to contain the growth of this microorganism. In this study, stable isotope labelling by amino acids in cell culture (SILAC) was used to conduct comparative proteomic quantitation of E. tarda ATCC 15947 under cold stress for two weeks. We identified 1391 proteins, of which 898 were quantifiable. Of these, 72 proteins were upregulated and 164 were downregulated in response to cold stress. Even though E. tarda ATCC 15947 is not a psychrophile, several key proteins related to DNA synthesis and transcription were significantly upregulated. Additionally, proteins related to haemolytic activities and gluconeogenesis were upregulated, even though E. tarda ATCC 15497 is considered non-virulent in aquaculture. This study therefore delineated the specific proteomic response of this E. tarda ATCC 15947 to prolonged cold stress.

Proteomic Analysis of Vibrio parahaemolyticus Under Cold Stress.

Vibrio parahaemolyticus is a kind of food-borne pathogenic bacterium, which can seriously infect food, especially seafood causing gastroenteritis and other disease. We studied the global proteome responses of V. parahaemolyticus under cold stress by nano-liquid chromatography-tandem mass spectrometry to improve the present understanding of V. parahaemolyticus proteomics events under cold stress. A total of 1151 proteins were identified and 101 proteins were differentially expressed, of which 69 were significantly up-regulated and 32 were downregulated. Functional categorization of these proteins revealed distinct differences between cold-stressed and control cells. These proteins were grouped into 21 functional categories by the clusters of orthologous groups (COG) analysis. The most of up-regulated proteins were functionally categorized as nucleotide transport and metabolism, transcription, function unknown, and defense mechanisms. These up-regulated proteins play an important role under cold stress.

[Studies and application method and quality control system for detecting hepatitis A virus in water for fruits and vegetables production].

OBJECTIVE: To explore detecting methods and quality control standard system for hepatitis A virus( HAV) in water for fruits and vegetables production. METHODS: 100 µL of coliphage MS2( 2. 5 × 10~9 pfu / mL) was added into each water sample, and positively charged membrane filter method was used to capture virus. The virus extraction efficiency of each sample can be calculated according to standard curve of coliphage. Then the quality control system and real-time RT-PCR method were established. RESULTS: This research compared the extraction efficiency of HAV and coliphage MS2 which were added into the water samples at the same time. The extraction efficiency of HAV was from 1. 24% to 32. 68%, and coliphage MS2 1. 64% to 14. 24%. The efficiency met the requirements of ISO / TS 15216-2-2013. Meanwhile, the HAV in 30 water samples were detected, and one was positive. The extraction efficiency of coliphage MS2 was from1. 24% to 24. 19%, and the standard deviation was 0. 0612. CONCLUSION: This research establish a quality control system to ensure the quality of test results.

Pre-Miocene birth of the Yangtze River.

The development of fluvial systems in East Asia is closely linked to the evolving topography following India-Eurasia collision. Despite this, the age of the Yangtze River system has been strongly debated, with estimates ranging from 40 to 45 Ma, to a more recent initiation around 2 Ma. Here, we present (40)Ar/(39)Ar ages from basalts interbedded with fluvial sediments from the lower reaches of the Yangtze together with detrital zircon U-Pb ages from sand grains within these sediments. We show that a river containing sediments indistinguishable from the modern river was established before ~23 Ma. We argue that the connection through the Three Gorges must postdate 36.5 Ma because of evaporite and lacustrine sedimentation in the Jianghan Basin before that time. We propose that the present Yangtze River system formed in response to regional extension throughout eastern China, synchronous with the start of strike-slip tectonism and surface uplift in eastern Tibet and fed by strengthened rains caused by the newly intensified summer monsoon.

Strain-level visualized analysis of cold-stressed Vibrio parahaemolyticus based on MALDI-TOF mass fingerprinting.

In this study, strain-level visualized analysis of cold-stressed Vibrio parahaemolyticus based on matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass fingerprinting was investigated. All the peptide mass fingerprinting profiles obtained were analyzed by self-organized map (SOM) and cluster analysis. Our results showed that the peptide mass fingerprinting profiles of V. parahaemolyticus substantially changed under cold stress at strain level. The cold-stressed V. parahaemolyticus strains were distributed to 14 neurons by SOM classification, almost totally different from the controls. This is the first time that so many strains had been chosen to study bacterial cold stress responses, which can help promote an overall understanding to stress responses of cold-stressed strains.

Proteomic analysis of protein expression in the induction of the viable but Nonculturable State of Vibrio harveyi SF1.

Vibrio harveyi has been reported to enter into a viable but nonculturable (VBNC) state. One marine V. harveyi strain, SF1 became nonculturable when incubated in seawater microcosm at 4 °C within 60 days. We investigated protein expression in the exponential phase of V. harveyi SF1 and compared it to the VBNC state. Cytosolic proteins were resolved by two-dimensional polyacrylamide gel electrophoresis using pH 4-7 linear gradients. Among these proteins, sixteen proteins which were strongly downregulated or upregulated in the VBNC cells were identified by MALDI-TOF-TOF mass spectrometry. The results indicated that the differentially expressed proteins were mainly focused on stress response proteins and key components of central and intermediary metabolism, like carbohydrate metabolism, transport, and translation. This study provided clues for understanding the mechanism of adaptation to the VBNC state.

[Identification of Vibrio metschnikovii from Homarus americanus and its changes in membrane fatty acid composition in response to low temperature].

OBJECTIVE: Bacterial strain F5-1 isolated from the Homarus americanus was characterized and its changes in membrane fatty acid composition in response to low temperature were also studied. METHODS: The physiological and biochemical characteristics were carried out by using VITEK 2 compact automated microbiology system. The 16S rRNA gene was sequenced and subjected to phylogenetic analysis. Fatty acids were detected by gas chromatography-mass spectrometry (GC-MS). RESULTS: Strain F5-1 was Gram-negative and susceptible to the vibriostatic agent O/129. Strain F5-1 was resistant to Penicillin. The isolated strain exhibited the highest levels of 99% probability to Vibrio metschnikovii based on the conventional physiological test. The sequence analysis of 16S rRNA gene of F5-1 isolation and comparison with that of other related vibrios showed that F5-1 was very close to V. metschnikovii (GenBank No. HQ658055). The similarity was 99%. The major fatty acids were C12:0, C14:0, C16:0 and C16:1 (n-7). Palmitoleic acid was the dominant unsaturated fatty acids. The major change in fatty acid composition occurred in response to low temperature, with an increase in palmitoleic acid from 34% to 40%. CONCLUSION: Bacterial strain F5-1 isolated from Homarus americanus was identified as V. metschnikovii and was sensitive to multiple drugs. The fatty acid composition of F5-1 was different from V. metschnikovii isolated from a drinking water reservoir near Vladivostok City in the Russia Far East. Results of this study indicated that environmental conditions allowed modulation of the fatty acid composition of V. metschnikovii.

Identification and characterization of the pathogenic effect of a Vibrio parahaemolyticus-related bacterium isolated from clam Meretrix meretrix with mass mortality.

A mass mortality of clam, Meretrix meretrix, occurred in Jiangsu Province of China in the late September of 2007. Of the isolates obtained from the diseased clams, MM21 had the strongest virulence to the clam in the virulence test, with a LD50 value of approximately 6 x 10(6)CFU ml(-1). MM21 was identified as Vibrio parahaemolyticus by the VITEK 2 Compact system and 16S rDNA sequencing. Detection of virulence-associated genes by PCR indicated that MM21 was positive for toxR and tlh, and negative for tdh. Compared with control group, histiocytes from MM21-infected clams displayed a variety of cytopathological changes by transmission electron microscopy examination, which included increased lipid droplets in hepatocytes, deposition of granules in the mantle, excessive secretion in the gill. The results of our study suggested that MM21 may have been an etiological element in the mass mortalities of hard clam (M. meretrix) in Jiangsu Province of China in 2007.

Potential Biochemical Mechanisms of Brain Injury in Diabetes Mellitus.

The goal of this review was to summarize current biochemical mechanisms of and risk factors for diabetic brain injury. We mainly summarized mechanisms published in the past three years and focused on diabetes induced cognitive impairment, diabetes-linked Alzheimer's disease, and diabetic stroke. We think there is a need to conduct further studies with increased sample sizes and prolonged period of follow-ups to clarify the effect of DM on brain dysfunction. Additionally, we also think that enhancing experimental reproducibility using animal models in conjunction with application of advanced devices should be considered when new experiments are designed. It is expected that further investigation of the underlying mechanisms of diabetic cognitive impairment will provide novel insights into therapeutic approaches for ameliorating diabetes-associated injury in the brain.

Proteomic analysis of Vibrio metschnikovii under cold stress using a quadrupole Orbitrap mass spectrometer.

Vibrio metschnikovii is a food-borne pathogen found in seafood worldwide. We studied the global proteome responses of V. metschnikovii under cold stress by nano-flow ultra-high-performance liquid chromatography coupled to a quadrupole Orbitrap mass spectrometer. A total of 2066 proteins were identified, among which 288 were significantly upregulated and 572 were downregulated. Functional categorization of these proteins revealed distinct differences between cold-stressed and control cells. Quantitative reverse transcription polymerase chain reaction analysis was also performed to determine the mRNA expression levels of seventeen cold stress-related genes. The results of this study should improve our understanding of the metabolic activities of cold-adapted bacteria and will facilitate a better systems-based understanding of V. metschnikovii.

Visualized analysis of cellular fatty acid profiles of Vibrio parahaemolyticus strains under cold stress.

Vibrio parahaemolyticus is a common foodborne bacterial pathogen, which survives in cold environments and is sometimes difficult to culture. Fatty acid analysis under cold stress was conducted for several V. parahaemolyticus strains using gas chromatography/mass spectrometry, and the results were compared with those of the controls. All the fatty acid profiles obtained were visualized by multidimensional scaling (MDS) and self-organized map (SOM). It was observed that the fatty acid profiles of V. parahaemolyticus substantially changed under cold stress. The percentage of methyl palmitate remarkably decreased and that of methyl palmitoleate (except for two strains) and methyl oleate increased. These findings demonstrate the role of fatty acids in cold stress. The changes in the fatty acid profiles illustrated by MDS and SOM could differentiate strains under cold stress from the controls and can potentially lead to a method of detecting injured cold-stressed V. parahaemolyticus.

Application of probabilistic neural network in bacterial identification by biochemical profiles.

An algorithm of Probabilistic Neural Network (PNN) for bacterial identification based on the probability matrix of API 20E system as a case study is reported. The PNN shows the correct identification of all the taxa belonging to the training and test sets and possesses merits over the conventional methods.

Adenovirus-vectored shRNAs targeted to the highly conserved regions of VP1 and 2B in tandem inhibits replication of foot-and-mouth disease virus both in vitro and in vivo.

Foot-and-mouth disease is a highly contagious and economically important disease of cloven-hoofed animals. RNA interference (RNAi) can be used as a rapid and specific antiviral approach. It was shown that treatment with recombinant adenovirus (Ad(VP1-2B)) carrying shRNAs targeted to the VP1 and 2B genes of FMDV expressed in tandem had marked antiviral effects against FMDV both in IBRS-2 cells and guinea pigs. Treatment with Ad(VP1-2B) both before and after FMDV infection was most effective in IBRS-2 cells, as the FMDV RNA transcripts could not be detected within 48 h post-challenge (hpc), and the viral RNA copy number at 72 hpc was only 0.02% of that in the positive control group. Delivery of Ad(VP1-2B) reduced significantly the susceptibility of guinea pigs to FMDV infection. All guinea pigs were protected within 3 days post challenge (dpc) when they were injected twice with the same dose of Ad(VP1-2B), and a third treatment with the same dose of Ad(VP1-2B) at 3 dpc was necessary to confer longer lasting protection (up to 6 dpc). In conclusion, application of such a adenovirus vector to inhibit more than one viral gene may be an advantageous method for prevention and therapy of FMDV infection.

Application of loop-mediated isothermal amplification for detection of Yersinia enterocolitica in pork meat.

A loop-mediated isothermal amplification (LAMP) method was developed for the detection of Yersinia enterocolitica isolates in both pure bacterial cultures and pork meat. The LAMP primers, which corresponded to the gyrB gene, accurately identified 4 different bioserotypes of Y. enterocolitica. These primers failed to detect Y. pseudotuberculosis, Y. frederiksenii, and 17 non-Yersinia strains. The sensitivity of the LAMP assay for the detection of Y. enterocolitica in pure culture was 65 CFU/mL (31.6 fg of genomic DNA). The LAMP assay was conducted for the detection of Y. enterocolitica strains in 21 pig tonsil samples and 73 pork meat samples obtained from 94 slaughtered pigs belonging to 4 different herds. Y. enterocolitica was found to be present in 4 tonsil samples and none in meat samples. This is the first report in which the LAMP assay was employed for the detection of Y. enterocolitica in food samples.

Rapid quantitative detection of Vibrio parahaemolyticus in seafood by MPN-PCR.

This study aimed to adopt MPN-PCR (most probable number-polymerase chain reaction) for rapid detection of the quantity of Vibrio parahaemolyticus in seafood. V. parahaemolyticus in seafood could be quantitated by MPN statistics according to PCR products. The sensitivity of MPN-PCR was 100 times higher than that of direct PCR. Of 225 seafood samples from Qingdao, 165 were positive for the presence of V. parahaemolyticus, with an MPN value of >719 per gram, and about 41.5% of samples were positive for tdh gene-possessing cells. Eighty muscle tissues from the 225 seafood samples were investigated by direct PCR and MPN-PCR, but no V. parahaemolyticus was detected. The MPN-PCR test could be completed in less than 16 h from the time of sample preparation. It was rapid, sensitive, and reliable for comprehensive detection and quick quantitative determination of V. parahaemolyticus in seafood and it revealed the potential risk of illness associated with their consumption.