Emergence of ehrlichiosis by a new tick-borne Ehrlichia species in China.

OBJECTIVES: From March to June 2021, the reported number of clinically diagnosed endemic typhus in Anhui and Hubei provinces of China nearly increased four-fold compared with the monthly average numbers in last 5 years. An etiological and epidemiological investigation was initiated. METHODS: The clinical specimens from the reported patients and the potential vector ticks were collected for molecular and serological detection, as well as cell culturing assay to identify the potential pathogen. RESULTS: Polymerase chain reaction and sequence analysis of rrs and groEL showed that the pathogen from these patients was Ehrlichia sp., isolated from Haemaphysalis longicornis attached to these patients. The phylogenetic analysis based on 39 Ehrlichia genomes suggested that it should be taxonomically classified as a novel species, tentatively named "Candidatus Ehrlichia erythraense". A total of 19 of 106 cases were confirmed as Candidatus Ehrlichia erythraense infections by polymerase chain reaction, sequencing, and/or serological tests. The most frequent symptoms were fever (100%), rashes (100%), asthenia (100%), anorexia (100%), and myalgia (79%). CONCLUSION: The occurrence of the disease presenting with fever and rashes in Anhui and Hubei provinces was caused by a novel species of the genus Ehrlichia; physicians need to be aware of this newly-discovered pathogen to ensure appropriate testing, treatment, and regional surveillance.

Environmental fate of microplastics in an urban river: Spatial distribution and seasonal variation.

Rivers are recognized as an important pathway for transport of microplastics (MPs) from land to sea, but limited information is available on the spatial distribution and seasonal variation of riverine MPs from upper reaches to estuaries. Such information is critical for source apportionment and development of effective management measures for riverine MPs. To fill the knowledge gap, we investigated the occurrence of MPs in surface water along an urban river in Guangzhou, southern China in wet and dry seasons. The abundances of MPs from 16 sampling sites in the wet and dry seasons varied from 0.123 to 1.84 particles m-3 and from 0.046 to 4.21 particles m-3, respectively. The spatial distribution of MP abundances showed an increasing trend from upstream to midstream and a decreasing trend from midstream to downstream and estuaries. The abundances of MPs peaked at the midstream, which is surrounded by a highly urbanized region with high population density (∼2530 persons per km2). The large surface water runoff during the wet season elevated the MP abundance in riverine water, except for that flowing through the central urban area where the abundance of MPs collected in the dry season was higher than that in the wet season. This was mainly ascribed to the large input from extensive anthropogenic activities and slow water flow rate in urban areas. The estimated monthly riverine MP fluxes from Humen, Hongqili, and Jiaomen were 7.42, 2.38, and 2.3 billion particles, respectively, in the wet season, and 0.86, 0.71, and 0.19 billion particles, respectively, in the dry season. An increase of riverine MP fluxes from Humen, Hongqili, and Jiaomen in the past three years was evident. The results from the present study provide valuable information for source apportionment of riverine MPs and support the initialization of possible MPs controlling measures.

The effect of Ditylum brightwellii (Bacillariophyceae) on colony development of bloom forming species Phaeocystis globosa (Prymnesiophyceae) under nutrient-replete condition.

To improve our knowledge of the factors regulating Phaeocystis globosa colony formation, the effects of the diatom Ditylum brightwellii on P. globosa colony development were investigated using co-culture and cell-free filtrate approaches. The co-culture experiments showed the moderate abundance of D. brightwellii significantly increased the number and size of colonies, whereas a dramatically decreased effect from high abundance of D. brightwellii. The low abundance of D. brightwellii promoted early formation of P. globosa colony. The cell-free filtrate experiments indicated that culture-filtrates from the exponential phase of D. brightwellii were stimulatory for P. globosa colony formation with more and bigger colonies formed, whereas an inhibitory effect from its senescence phase filtrates. D. brightwellii may influence P. globosa colony formation by regulating the growth of P. globosa solitary cells. Our results suggest that D. brightwellii influences P. globosa colony development, but its effects vary according to its concentrations and growth phases.

Genetic Diversity, Antimicrobial Resistance, and Virulence Genes of Aeromonas Isolates from Clinical Patients, Tap Water Systems, and Food.

OBJECTIVE: This study aimed to evaluate the genetic diversity, virulence, and antimicrobial resistance of Aeromonas isolates from clinical patients, tap water systems, and food. METHODS: Ninety Aeromonas isolates were obtained from Ma'anshan, Anhui province, China, and subjected to multi-locus sequence typing (MLST) with six housekeeping genes. Their taxonomy was investigated using concatenated gyrB-cpn60 sequences, while their resistance to 12 antibiotics was evaluated. Ten putative virulence factors and several resistance genes were identified by PCR and sequencing. RESULTS: The 90 Aeromonas isolates were divided into 84 sequence types, 80 of which were novel, indicating high genetic diversity. The Aeromonas isolates were classified into eight different species. PCR assays identified virulence genes in the isolates, with the enterotoxin and hemolysin genes act, aerA, alt, and ast found in 47 (52.2%), 13 (14.4%), 22 (24.4%), and 12 (13.3%) of the isolates, respectively. The majority of the isolates (≥ 90%) were susceptible to aztreonam, imipenem, cefepime, chloramphenicol, gentamicin, tetracycline, and ciprofloxacin. However, several resistance genes were detected in the isolates, as well as a new mcr-3 variant. CONCLUSIONS: Sequence type, virulence properties, and antibiotic resistance vary in Aeromonas isolates from clinical patients, tap water systems, and food.

Ginkgolic acid (GA) suppresses gastric cancer growth by inducing apoptosis and suppressing STAT3/JAK2 signaling regulated by ROS.

Gastric cancer is a frequently occurring cancer with high mortality each year worldwide. Finding new and effective therapeutic strategy against human gastric cancer is still urgently required. Ginkgolic acid (GA), a botanical drug, is extracted from the seed coat of Ginkgo biloba L. with various bioactive properties, including anti-tumor. Unfortunately, if GA has antitumor effect on human gastric cancer and the underlying molecular mechanisms have yet to be investigated. In the present study, we found that GA markedly reduced the gastric cancer cell viability. Furthermore, GA treatment led to the reduced migration ability of gastric cancer cells, which was associated with the decreased protein expression levels of Rho-associated protein kinase 1 (ROCK1), matrix metalloproteinase-2 (MMP-2), MMP-9 and α-smooth muscle actin (α-SMA). In addition, GA dose-dependently induced apoptosis in gastric cancer cells through activating Caspase-9/-3 and poly(ADP-Ribose) polymerase (PARP), which was along with the reduced Bcl-2 and Bcl-xl expression levels, and the elevated Bax and Bad levels. Consistently, Cyto-c protein expression in cytoplasm was also up-regulated by GA. Moreover, the production of reactive oxygen species (ROS) was significantly induced by GA. The activation of signal transducer and activator of transcription 3/janus kinase 2 (Stat3/JAK2) signaling pathway was inhibited by GA treatment. Intriguingly, blocking Stat3/JAK2 activation could further promote apoptosis and reduce cell viability induced by GA. However, GA-induced cell death was clearly abolished by ROS scavenger NAC, while the activation of Stat3/JAK2 signaling was restored by NAC. In vivo, GA showed effective role in reducing gastric tumor growth. Together, the findings here indicated that GA could be considered as an effective therapeutic candidate against human gastric cancer progression in future.

Using a mixture of wastewater and seawater as the growth medium for wastewater treatment and lipid production by the marine diatom Phaeodactylum tricornutum.

This study was conducted to evaluate the potential of the marine diatom Phaeodactylum tricornutum in nutrient removal coupled with biodiesel production using different ratios of mixed municipal wastewater (MW) and seawater (SW) as the growth medium. The results indicated that P. tricornutum exhibited high nutrient removal efficiency with the ratios of MW: SW = 1:1 and MW: SW = 2:1, e.g. 87.7-89.9% for chemical oxygen demand (COD), 82.2-86.7% for total nitrogen (TN), 96.0-97.0% for total phosphorus, and 76.9-84.2% for ammonium (NH3-N). Significantly higher biomass and lipid productivity were obtained with aeration. The highest lipid productivity of P. tricornutum was 54.76 mg/L/day, which was obtained with a two-step cultivation using the ratio of MW: SW = 1:1 by diluting half of the mixture and bubbling with 5% CO2 during the second step. These results suggested that the marine diatom P. tricornutum exhibited great potential for using mixed wastewater for wastewater treatment and biodiesel production.

Effects of fundamental nutrient stresses on the lipid accumulation profiles in two diatom species Thalassiosira weissflogii and Chaetoceros muelleri.

Microalgae are considered as attractive feedstocks for biofuel production nowadays because of their high lipid contents and easy cultivation. In the present study, two diatoms, Thalassiosira weissflogii and Chaetoceros muelleri, were cultured under various nutrient-limitation conditions to explore their comprehensive lipid accumulation profiles for further commercialization. In T. weissflogii, the highest neutral lipid accumulation and highest lipid productivity (14.28 mg L-1 day-1) were both recorded under P-limitation. In C. muelleri, the highest lipid content (35.03% of dry cell weight), highest neutral lipid accumulation, and highest lipid productivity (29.07 mg L-1 day-1) were all recorded under N-limitation. Besides, the predominant fatty acids of T. weissflogii and C. muelleri were myristic acid (C14:0), palmitic acid (C16:0), and palmitoleic acid (C16:1), with the amounts of 58.4-74.4 and 74.1-87.7% of the total fatty acids, respectively. Moreover, nutrient limitations led to a lower proportion of polyunsaturated fatty acids (PUFA) than that of saturated fatty acid (SFA) and monounsaturated fatty acid (MUFA) in both species. The ratios of (SFA + MUFA) to PUFA were from 1.65 to 3.01 in T. weissflogii, and up to 3.61 to 8.59 in C. muelleri. Our results suggested the feasibility of C. muelleri as biodiesel feedstock due to its more suitable fatty acid composition and higher lipid productivity compared to T. weissflogii.

Differential cellular responses associated with oxidative stress and cell fate decision under nitrate and phosphate limitations in Thalassiosira pseudonana: Comparative proteomics.

Diatoms are important components of marine ecosystems and contribute greatly to the world's primary production. Despite their important roles in ecosystems, the molecular basis of how diatoms cope with oxidative stress caused by nutrient fluctuations remains largely unknown. Here, an isobaric tags for relative and absolute quantitation (iTRAQ) proteomic method was coupled with a series of physiological and biochemical techniques to explore oxidative stress- and cell fate decision-related cellular and metabolic responses of the diatom Thalassiosira pseudonana to nitrate (N) and inorganic phosphate (P) stresses. A total of 1151 proteins were detected; 122 and 56 were significantly differentially expressed from control under N- and P-limited conditions, respectively. In N-limited cells, responsive proteins were related to reactive oxygen species (ROS) accumulation, oxidative stress responses and cell death, corresponding to a significant decrease in photosynthetic efficiency, marked intracellular ROS accumulation, and caspase-mediated programmed cell death activation. None of these responses were identified in P-limited cells; however, a significant up-regulation of alkaline phosphatase proteins was observed, which could be the major contributor for P-limited cells to cope with ambient P deficiency. These findings demonstrate that fundamentally different metabolic responses and cellular regulations are employed by the diatom in response to different nutrient stresses and to keep the cells viable.

Seasonal variations of phytoplankton assemblages and its relation to environmental variables in a scallop culture sea area of Bohai Bay, China.

Seasonal variations of phytoplankton assemblages were examined in a scallop culture sea area of Bohai Bay (China) with regard to some major physical and chemical variables. Samples were collected at three stations from July 2011 to September 2013. A total of 134 species belong to 4 phyla were identified, of which 104 were diatoms, 27 were dinoflagellates, 1 was euglenophyte and 2 were chrysophytes. The cells abundance in autumn (55.44×103cells/L) was higher than that in summer (6.99×103cells/L), spring (3.46×103cells/L) and winter (2.69×103cells/L). The Shannon-Wiener diversity index was higher in summer (3.06), followed by spring (3.02) and winter (2.91), and low in autumn (1.40). Results of canonical correspondence analysis showed that phosphate, salinity, temperature, silicate and DIN/SiO2 ratio were the most important environmental factors influencing the variation of phytoplankton community structure. It is suggested that eutrophication resulted from scallop culture would cause a potential red tide risk.

Cellular responses associated with ROS production and cell fate decision in early stress response to iron limitation in the diatom Thalassiosira pseudonana.

Investigation of how diatoms cope with the rapid fluctuations in iron bioavailability in marine environments may facilitate a better understanding of the mechanisms underlying their ecological success, in particular their ability to proliferate rapidly during favorable conditions. In this study, using in vivo biochemical markers and whole-cell iTRAQ-based proteomics analysis, we explored the cellular responses associated with reactive oxygen species (ROS) production and cell fate decision during the early response to Fe limitation in the centric diatom Thalassiosira pseudonana. Fe limitation caused a significant decrease in Photosystem (PS) II photosynthetic efficiency, damage to the photosynthetic electron transport chain in PS I, and blockage of the respiratory chain in complexes III and IV, which could all result in excess ROS accumulation. The increase in ROS likely triggered programmed cell death (PCD) in some of the Fe-limited cells through synthesis of a series of proteins involved in the delicate balance between pro-survival and pro-PCD factors. The results provide molecular-level insights into the major strategies that may be employed by T. pseudonana in response to Fe-limitation: the reduction of cell population density through PCD to reduce competition for available Fe, the reallocation of intracellular nitrogen and Fe to ensure survival, and an increase in expression of antioxidant and anti-PCD proteins to cope with stress.

Biomass, total lipid production, and fatty acid composition of the marine diatom Chaetoceros muelleri in response to different CO2 levels.

The marine diatom Chaetoceros muelleri grown under air (0.03% CO2), 10%, 20%, and 30% CO2 conditions was evaluated to determine its potential for CO2 reduction coupled with biodiesel production. The results indicated that C. muelleri grows well with high CO2 aeration levels (10-20%) and is induced to accumulate lipids under 10-30% CO2. In particular, the highest values of the maximum biomass concentration (1.059gL(-1)), maximum specific growth rate (0.868d(-1)), maximum biomass productivity (0.272gL(-1)d(-1)), maximum CO2 biofixation (0.428gL(-1)d(-1)), and total lipid (43.40% dry weight) and neutral lipid contents were all obtained with 10% CO2 aeration. Moreover, the analysis of the fatty acid composition of C. muelleri revealed the predominance of C14-C18 fatty acids (>90%) and saturated and monounsaturated fatty acids (>80%) under all CO2 levels. The results suggest that C. muelleri has great potential to biodiesel production using flue gases.

Cellular metabolic responses of the marine diatom Pseudo-nitzschia multiseries associated with cell wall formation.

In this study a comparative proteomics approach involving a mass spectrometric analysis of synchronized cells was employed to investigate the cellular-level metabolic mechanisms associated with siliceous cell wall formation in the pennate diatom Pseudo-nitzschia multiseries. Cultures of P. multiseries were synchronized using the silicate limitation method. Approximately 75% of cells were arrested at the G2+M phase of the cell cycle after 48 h of silicate starvation. The majority of cells progressed to new valve synthesis within 5h of silicon replenishment. We compared the proteome of P. multiseries at 0, 4, 5, and 6h of synchronization progress upon silicon replenishment using two-dimensional gel electrophoresis. Forty-eight differentially expressed protein spots were identified in abundance (greater than two-fold change; P<0.005), some of which are predicted to be involved in intracellular trafficking, cytoskeleton, photosynthesis, lipid metabolism, and protein biosynthesis. Cytoskeleton proteins and clathrin coat components were also hypothesized to play potential roles in cell wall formation. The proteomic profile analysis suggests that P. multiseries most likely employs multiple synergistic biochemical mechanisms for cell wall formation. These results improve our understanding of the molecular mechanisms underlying silicon cell wall formation and enhance our understanding of the important role played by diatoms in silicon biogeochemical cycling.

iTRAQ-based proteomic analysis of the metabolism mechanism associated with silicon response in the marine diatom Thalassiosira pseudonana.

Silicon is a critical element for diatom growth; however our understanding of the molecular mechanisms involved in intracellular silicon responses are limited. In this study, an iTRAQ-LC-MS/MS quantitative proteomic approach was coupled with an established synchrony technique to reveal the global metabolic silicon-response in the model diatom Thalassiosira pseudonana subject to silicon starvation and readdition. Four samples, which corresponded to the time of silicon starvation, girdle band synthesis, valve formation, and right after daughter cell separation (0, 1, 5, 7 h), were collected for the proteomic analysis. The results indicated that a total of 1,831 proteins, representing 16% of the predicted proteins encoded by the T. pseudonana genome, could be identified. Of the identified proteins, 165 were defined as being differentially expressed proteins, and these proteins could be linked to multiple biochemical pathways. In particular, a number of proteins related to silicon transport, cell wall synthesis, and cell-cycle progress could be identified. In addition, other proteins that are potentially involved in amino acid synthesis, protein metabolism, and energy generation may have roles in the cellular response to silicon. Our findings provide a range of valuable information that will be of use for further studies of this important physiological response that is unique to diatoms.

[An etiological survey on a foodborne disease epidemic outbreak caused by Salmonella enteritidis].

OBJECTIVE: To conduct an etiological molecular epidemiological survey and laboratory test on a foodborne disease epidemic outbreak to make clear of the cause and implement effective prevention and control on it. METHODS: On May 12th 2012, 135 kindergarten children were sent to Xuzhou City People's Hospital and Children's Hospital with gastrointestinal infection disease. A total of 34 anus swab samples and 4 vomit samples were collected from the patients. Real-time PCR rapid detection, strains separation and cultivation, phage lysis experiments, ATB automated identification system were used to make etiological detection and identification. The genomic DNA of salmonella enteritidis were typed with the pulsed-field gel electrophoresis (PFGE), cluster analysis were carried out together with the patterns of local Salmonella infections. RESULTS: Children in 20 classes were suffered from the gastrointestinal infection among the 21 classes. There were no significant aggregation of class distribution. Among the 135 patients, 76 were boys (56.3%) and 59 were girls (43.7%). The main symptoms were fever (above 38°C), diarrhea and bellyache. Through real-time PCR detection and strains separation, 19 salmonella enteritidis were isolated from 34 anus swab samples of suspected cases and the detection rate was 56%. There were no strains detected from vomit samples. All of the 19 salmonella enteritidis showed the same serological subtype, biochemical reaction, drug sensitivity and phage lysis pattern. The salmonella enteritidis had the identical PFGE pattern (100% similarity), and were different from the pattern of local sporadic infection cases. CONCLUSION: It was confirmed that this was an epidemic outbreak of foodborne disease caused by homologous salmonella enteritidis by epidemiological survey, clinical information, lab etiological test and molecular typing.

[Analysis of the variation and changes of Yersinia enterocolitica in Ningxia area from 1984 to 2011].

OBJECTIVE: To analyze the genetic evolution and bacterial type changes of Yersinia enterocolitica in the Ningxia area between year 1984 and 2011. METHODS: A total of 296 strains of Yersinia enterocolitica was collected from diarrhea patients, pig, rodents, sheep and dogs between year 1984 and 2011. The serotype, biotype, ail, ystA, ystB, yadA, virF and other toxic genes were detected. The PFGE subtypes of serotype O:3 and O:9 strains and the cluster features were analyzed. RESULTS: Out of 296 Yersinia enterocolitica strains, pig was the main host, accounting for 65.20% (193/296), followed by rodents, accounting for 32.43% (96/296). Serotype and biotype had their own respective dominant types in different periods. During 1984 and 1985, 2 strains of serotype O:3 and 3 strains of serotype O:9 were isolated, all belonged to biotype 3. Because of lack of strains, there were no obvious dominant types found. Between 1997 and 1999, 177 strains of serotype O:9 Yersinia enterocolitica were isolated as the dominant strain; and there were 178 strains of biotype 2 Yersinia enterocolitica were found. During 2007 and 2011, 54 strains of serotype O:3 Yersinia enterocolitica were isolated as dominant strain; followed by 26 strains of serotype O:5. There were separately 44 and 59 strains of biotype 1A and biotype 3. The PCR test divided the 248 strains into 4 types, including pathogenic strains as type I (ail(+), ystA(+), ystB(-), yadA(+), virF(+)). The PFGE divided the serotype O:3 into 12 types, in which K6GN11C30021 and K6GN11C30012 were the dominant types, accounting for 63.64% (42/66). The serotype O:9 were divided into 14 types, in which K6GN11C90010, K6GN11C90008, K6GN11C30018 and K6GN11C90003 were the dominant types, accounting for 89.01% (162/182). CONCLUSION: The different serotypes of isolated strains in Ningxia district showed different dominant bacteria in different periods; while the biotypes also changed with serotypes. The Yersinia enterocolitica isolated from different years showed great variation.