Novel breeding approach for Japanese flounder using atmosphere and room temperature plasma mutagenesis tool.

BACKGROUND: Artificial induction of mutagenesis is effective for genetic resource innovation and breeding. However, the traditional mutation methods for fish breeding are not convenient or safe for daily use. Hence, development of a simple, safe and effective mutagenesis method with a high mutation rate and applicability to multiple fish species, is needed. RESULTS: We reported the first successful mutagenesis in a marine aquaculture fish species, Japanese flounder, Paralichthys olivaceus, using a novel atmosphere and room temperature plasma (ARTP) mutagenesis tool. ARTP treatment time was optimized for the fertilized eggs and sperm, respectively. Eggs fertilized for 60 min were treated by ARTP with a radio-frequency power input of 120 W, and the ARTP treatment time was 25 min. Under an ARTP radio-frequency power input of 200 W, the optimal treatment time for sperm diluted with Ringer's solution by 1:40 v/v was 10 min. The ARTP-treated group presented differences in morphological traits such as body height, total length among individuals at day 90 after hatching. Whole-genome sequencing was used to reveal the mutation features of ARTP-treated individuals collected at day 120 after hatching. In total, 69.25Gb clean data were obtained from three controls and eight randomly selected ARTP-treated individuals, revealing 240,722 to 322,978 SNPs and 82,149 to 86,798 InDels located in 17,394~18,457 and 12,907~13,333 genes, respectively. The average mutation rate reached 0.064% at the genome level. Gene ontology clustering indicated that genes associated with cell components, binding function, catalytic activity, cellular process, metabolic process and biological regulation processes had higher mutation rates. CONCLUSIONS: ARTP mutagenesis is a useful method for breeding of fish species to accelerate the selection of economically important traits that would benefit the aquaculture industry, given the variety of mutations detected.

Heat and light stresses affect metabolite production in the fruit body of the medicinal mushroom Cordyceps militaris.

Cordyceps militaris is a highly valued edible and medicinal fungus due to its production of various metabolites, including adenosine, cordycepin, N6-(2-hydroxyethyl)-adenosine, and carotenoids. The contents of these metabolites are indicative of the quality of commercially available fruit body of this fungus. In this work, the effects of environmental abiotic factors, including heat and light stresses, on the fruit body growth and metabolite production in C. militaris were evaluated during the late growth stage. The optimal growth temperature of C. militaris was 20 °C. It was found that a heat stress of 25 °C for 5-20 days during the late growth stage significantly promoted cordycepin and carotenoid production without affecting the biological efficiency. Light stress at 6000 lx for 5-20 days during the late growth stage significantly promoted cordycepin production but decreased the carotenoid content. Both heat and light stresses promoted N6-(2-hydroxyethyl)-adenosine production. In addition, gene expression analysis showed that there were simultaneous increases in the expression of genes encoding a metal-dependent phosphohydrolase (CCM_04437) and ATP phosphoribosyltransferase (CCM_04438) that are involved in the cordycepin biosynthesis pathway, which was consistent with the accumulation of cordycepin during heat stress for 5-20 days. A positive weak correlation between the cordycepin and adenosine contents was observed with a Pearson correlation coefficient of 0.338 (P < 0.05). The results presented herein provide a new strategy for the production of a superior quality fruit body of C. militaris and contribute to further elucidation of the effects of abiotic stress on metabolite accumulation in fungi.

Suitability of coconut bran and biochar as a composite substrate for lettuce cultivation in aquaponic systems.

Growth substrates are essential for aquaponic systems and play an important role in vegetable growth and water quality. In this study, we explored an innovative combination of coconut bran and coconut shell biochar (CSB) as a composite growth substrate for lettuce cultivation in aquaponic systems. The study included the control (100 % coconut bran as the growth substrate) and treatment groups (T1-T5; containing 10 %, 20 %, 30 %, 40 %, and 50 % CSB as the growth substrate, respectively). The substrate properties; lettuce growth performance; and soil enzyme activity, nitrogen content, and abundance of microbial communities in the substrate were analyzed to determine the optimal substrate. Our findings indicated that CSB incorporation significantly altered the properties of the substrate, resulting in increased dry and bulk densities, pH, and water-holding capacity, and decreased electrical conductivity, water-absorption capacity, and porosity. Furthermore, the fresh weight of lettuce was notably increased in the treatment groups. The activities of fluorescein diacetate hydrolase, urease, nitrate reductase, and hydroxylamine reductase initially increased and further decreased, reaching the maximum in the T3 group. Conversely, the activity of nitrite reductase and contents of available nitrogen, nitrate-nitrogen, and ammonium-nitrogen in the substrates initially decreased and further increased, with the minimum values observed in the T3 group. The microbial sequencing results indicated that CSB incorporation significantly increased the microbial diversity and relative abundance of microorganisms associated with nitrogen transformation. Moreover, 30 % CSB incorporation exhibited the greatest effect on lettuce growth, with a 34.5 % and 31.6 % increase in fresh weight compared to the control during the growth and harvest periods, respectively. This study indicated the enormous potential of biochar in the research and development of green technologies for substrate amendment in aquaponic systems.

Soil fertility, salinity and nematode diversity influenced by Tamarix ramosissima in different habitats in an arid desert oasis.

The aim of this paper was to assess the influence of tamarisk shrubs on soil fertility, salinity and nematode communities in various habitats located in an arid desert-oasis region in northwest China. Three habitats were studied: sand dune, riparian zone and saline meadow, where tamarisk shrubs have been established in recent decades in order to vegetation restoration used as desertification control and saline land rehabilitation projects and become the dominant plant community. The parameters measured include soil organic carbon (SOC), total nitrogen, available phosphorus (P) and potassium (K), pH, salt component, and nematode community characteristics. Enrichment ratios (a comparison of the soil measurements between soils under canopy and in the open interspaces) for soil nutrients and salinity were used to evaluate fertility and salinity islands underneath the tamarisk shrubs. The soil nematode community was used as a biological indicator of soil condition. SOC and available P and K were higher beneath the plant canopy than in the open interspaces outside that canopy. The enrichment ratios for SOC and nutrients were highest for the sand dune habitat and tamarisk shrubs clearly created islands of greater salinity under the canopies. Nematode abundance per 100 g dry soil varied considerably between the locations and habitats, with the highest abundance found in sand dune and the lowest in saline meadow. A significantly higher nematode abundance and a lower trophic diversity were found in soils under the canopy compared to the soils in the open interspaces. With the exception of saline meadow, the abundance of bacterivores increased and fungivores decreased under the canopy relative to the open interspaces, and bacterivores dominated under the canopies in the sand dune and riparian habitats. The enrichment ratios for salinity were higher than for fertility, suggesting that improved soil fertility can not limit the impact of salinization beneath tamarisk shrubs. The adverse effect of salt accumulation on the soil environment should be taken into account when using tamarisk as restoration plant species, especially in saline meadow and controlling of tamarisk density should be considered when undertaking re-vegetation projects in the arid desert oasis regions.

GbWRKY1, a novel cotton (Gossypium barbadense) WRKY gene isolated from a bacteriophage full-length cDNA library, is induced by infection with Verticillium dahliae.

WRKY transcription factor proteins play important roles in diverse stress responses. In this study, we first cloned a novel WRKY from our constructed bacteriophage full-length cDNA library for cotton (Gossypium barbadense). The plants were stressed by exposure to a defoliating strain of Verticillium dahliae. The capacity of primary cDNA library was 1.28 x 10(6) PFU and the titer of the amplified cDNA library was > 10(10) PFU mL(-1). The recombination rate of the library was 94% and average insert size was about 1.1 kb. This novel gene, named GbWRKY1 was 1971 bp long and encodes a protein of 489 amino acids. It contains two characteristic WRKY domains and two zinc finger motifs. The sub-cellular assay indicated that GbWRKY1-GFP fusion protein was localized in the nucleus. Furthermore, Northern blot analysis showed that expression pattern of GbWRKY1 was similar among tissue types (roots, stems and leaves), but differed between pathogen-infiltrated and Czapek medium-infiltrated (untreated control) plants. Quantitative real-time PCR showed that GbWRKY1 could also be induced by salicylic acid (SA), methyl jasmonate (MeJA) and 1-aminocyclopropane-1-carboxylic acid (ACC). These findings clearly suggest that as a pathogen-inducible transcription factor GbWRKY1 plays an important role in plant defense responses.

PRNP mutations in a series of apparently sporadic neurodegenerative dementias in China.

Mutations in prion protein gene (PRNP) may lead to genetic prion disease, which usually has a broad range of phenotypic presentations that overlap with other neurodegenerative dementias. In this study, we screened the PRNP gene to evaluate the frequency of PRNP mutations and their correlations with clinical phenotype in 185 sporadic neurodegenerative dementia cases and 310 control subjects. Samples of DNA from each subject underwent polymerase chain reaction (PCR) amplification and direct sequencing of PRNP. The clinical characteristics of patients carrying PRNP mutations were detailed. We identified five different PRNP mutations in five patients, of which three were novel (S97N, F198V, and R208C) and two were known (D178N-129M and M232R). The rate of PRNP mutation was 2.70% in our sample. Though future studies confirming the correlation between PRNP mutations and clinical phenotype need to be undertaken, PRNP genotyping may be a valuable tool to differentiate between prion disease and other neurodegenerative dementias.

Effect of HNO3 concentration on a novel silica-based adsorbent for separating Pd(II) from simulated high level liquid waste.

A new kind of silica-based (Crea + TODGA)/SiO2-P adsorbent with high selectivity adsorption for palladium (Pd) was synthesized to examined the applicability for partitioning process of high level liquid waste (HLLW). Adsorption behavior of Pd(II) towards (Crea + TODGA)/SiO2-P adsorbent and stability of adsorbent against HNO3 solution were investigated by batch method. The degradation parts of (Crea + TODGA)/SiO2-P dissolved in liquid phase were estimated by total organic carbon (TOC) analyzer. (Crea + TODGA)/SiO2-P adsorbent showed good selectivity adsorption for Pd(II) and reached equilibrium within 24 hr. The adsorption ability of (Crea + TODGA)/SiO2-P for Pd(II) and the content of TOC leaked decreased with the increasing of HNO3 concentration. In 3 M HNO3, the average of K d values were 85.03 cm3/g and 26.10 cm3/g after contact time one to 28 days at 298 K and 323 K, respectively. While the content of TOC leaked from the adsorbent after 28 days were 1095 ppm (298 K) and 2989 ppm (323 K), respectively. Therefore, the adsorbent showed good stability at 298 K after contact with nitric acid for a long time. All results indicated (Crea + TODGA)/SiO2-P can be proposed as an applicable and efficient absorbent for separation of Pd(II) in 3 M HNO3 at 298 K.

[Surveillance of schistosomiasis transmission risk in Jingmen City, Hubei Province].

OBJECTIVE: To investigate the risk factors of schistosomiasis transmission in Jingmen City. METHODS: The Oncomelania hupensis snails, the wild animal feces, and infection source were selected as the monitoring objects to carry out the schistosomiasis risk monitoring. I-III levels of risk environments were treated with appropriate measures. RESULTS: A total of 52 environments and three water systems were monitored and 1 542 snails were dissected but no Schistosoma infected snails were found. Nine fecal samples were collected from the areas with snails, and no eggs of Schistosoma were found. Eighty-nine samples of cattle/sheep faces, and mice and dogs were collected, and three samples of cattle feces were found with Schistosoma eggs. Five environments were assessed as Grade II, and 48 environments were assessed as Grade III, and 2 environments were assessed as no risk of schistosomiasis transmission. CONCLUSIONS: In Jingmen City, the mollusciciding work from May to June could decrease the density of snails and the risk of schistosomiasis transmission efficiently. The schistosome-infected cattle were the main infection source, and therefore, the cattle and snails should be administrated simultaneously.

[Schistosomiasis surveillance among migrant population in Haining City in 2015].

OBJECTIVE: To understand the schistosomiasis epidemic situation among migrant population, so as to assess the potential risk of schistosomiasis transmission in Haining City. METHODS: Five districts in Haining City were randomly selected to investigate the schistosomiasis endemic situation in migrant population from the schistosomiasis endemic provinces. RESULTS: Totally 1 207 persons were investigated and four positives were found in serological tests, with the positive rate of 0.33%, but no patients were found in pathologic tests. The positive cases in serological tests were mainly between 31-40 years old, with the positive rate of 1.68%, which was higher than that of other age groups (χ2 = 6.590, P < 0.05). CONCLUSIONS: The schistosomiasis surveillance work in migrant population still should be strengthened in Haining City.

[Statistics of causes of death and analysis of risk factors in a surgical intensive care unit].

OBJECTIVE: To summarize the causes of death and to analyze the risk factors in a surgical intensive care unit (SICU). METHODS: The relevant information of patients died in the SICU of Xijing Hospital of Fourth Military Medical University in past 15 years (from December 1999 to February 2015) was retrospectively analyzed. The gender, age, reason and date of hospitalization, date of transfer SICU, past medical history, whether or not admitted directly from emergency department or transferred from other department, operated or not, date of death, the main cause of death, acute physiology and chronic health evaluation II (APACHE II) score, the history of undergoing mechanical ventilation, continuous renal replacement therapy (CRRT), or antifungal therapy, as well as the ratio of the patients with body temperature higher than 39 °C, white blood cell (WBC) count higher than 10 x 109/L, platelet (PLT) count below 100 x 109/L, albumin (Alb) below 35 g/L of two periods, namely from December 1999 to July 2007 (the first period), and from August 2007 to February 2015 (the second period) were compared. The above parameters were compared with those of 201 survivors in SICU, and the risk factors leading to death were analyzed by logistic regression. RESULTS: From December 1999 to February 2015, 4 317 patients were taken care of in the SICU. Among them, the number of death was 186, and the mortality rate was 4.3%. In the first time period (from December 1999 to July 2007), the total number of patients was 1 356, and the number of death were 109 (the mortality rate was 8.0%). In the second period, i.e. from August 2007 to February 2015, the number of SICU patients was 2,961, and 77 died (the mortality rate was 2.6%). The difference of mortality rate between the two periods was statistically significant (χ² = 66.707, P = 0.001 ). The death rate of patients transferred directly from emergency department in the first period was 79.8% (87/109), and it was lower in the second period (51.9%, 40/77, χ² = 16.181, P = 0.001 ). The death rate of the patients with blood A1b below 35 g/L in the second period (59.7%, 46/77) was higher than that of the first period (41.3%, 45/109, χ² = 6.151, P = 0.017). The top three causes of death from December 1999 to February 2015 were sepsis (38.2%), trauma (16.7%), and operation for cancer (14.0%). In the first period, the top three causes of death were sepsis (35.8%), trauma (22.0%), and operation for cancer (13.8%). In the second period, the top three causes of death were sepsis (41.6%), damage of the central nervous system (16.9%), and operation for cancer (14.3%). Top three reasons for SICU admission were trauma (29.03%), abdominal pain (20.97%) and other reasons (18.82%). Top three departments from which the patients were transferred were the emergency department (19.35%), orthopedics department (17.20%), and hepatobiliary department (16.13%). Logistic regression analysis showed that age [odds ratio (OR) = 2.025, 95% confidence interval (95% CI) = 1.500-2.734, P = 0.000], mechanical ventilation (OR = 3.514, 95% CI = 1.701-7.259, P = 0.001), CRR T (OR = 5.604, 95% CI = 3.003-10.459, P = 0.000 ), body temperature higher than 39 °C (OR = 1.992, 95%CI = 1.052-3.771, P = 0.034) were the risk factors of death in SICU patients. CONCLUSION: Sepsis and severe trauma are the leading causes of death in severe SICU patients, to whom with risk factors of death enough attention should be given.

[Endemic situation of schistosomiasis in a surveillance site of river and lake regions in Haining City, 2012-2014].

OBJECTIVE: To understand the endemic dynamics and situation of schistosomiasis in a provincial surveillance site in Haining City, Zhejiang Province. METHODS: The Oncomelania hupensis snail status, schistosome infection situations of permanent residents and floating population in Qinmin Village, Haining City were monitored from 2012 to 2014 according to The Surveillance Programs of Schistosomiasis in Surveillance Sites of Zhejiang Province. RESULTS: Totally 600 permanent residents were examined from 2012 to 2014. The total positive rate of the serum antibody against Schistosoma japonicum was 3.17% (19/600). The rates in each year were 2.50% (5/200), 0 (0/200), and 7.00% (14/200), respectively, and the rate in 2014 were significantly higher than that in 2013 (χ2 = 14.508, P < 0.01), but no positives were found in the fecal examination. Totally 1 591 floating population were involved in the surveillance, the positive rate of serum antibody were 0.94% (15/1 591). The rates in active surveillance and passive surveillance were 0.92% (14/1 521) and 1.43% (1/70), respectively, and there were no statistically significant difference between them (χ2 = 1.659, P > 0.05). The positive rates of serum antibody among each year were 1.50% (8/534), 0.56% (3/531), and 0.76% (4/526), respectively, and the difference was not statistically significant (χ2 = 2.752, P >0.05), but no positives were found in the fecal examination. The snail surveillance showed that there were no environments with infected snails and imported snails. CONCLUSIONS: There exist the infection sources of schistosomiasis in Haining City. Therefore, we should be on high alert for the potential of local endemic of the disease. Meanwhile, we still should strengthen the snail surveillance as well as the infection surveillance among the floating population.

Blocking abdominal lymphatic flow attenuates acute hemorrhagic necrotizing pancreatitis -associated lung injury in rats.

BACKGROUND: Acute hemorrhagic necrotizing pancreatitis (AHNP) is a severe acute inflammatory of pancreas that can lead to extrapancreatic organ disfunction. The lung and intestine is the most common involved organs, and abdominal lymphatic flow may contribute to AHNP- associated organ injury. In this study, we investigated the impact of thoracic duct ligation and drainage on lung and intestine injury in rats with AHNP. METHODS: Thirty-two male Wistar rats were randomly divided into 4 groups: sham operation group, AHNP group, AHNP + ligation group and AHNP + drainage group. Rat AHNP model was induced by retrograde injection of 3.5% sodium deoxycholate into the biliopancreatic duct, and the sham operation group was injected only with saline. In AHNP + ligation group and AHNP + drainage group, thoracic duct was ligated or drainaged before model induction. At 6 h after model induction, the bronchoalveolar lavage fluid (BALF) were collected for determination of tumor necrosis factor-alpha (TNF-alpha), and the tissues of lung, intestine and pancrease were harvested individually for pathohistological evaluation and the myeloperoxidase (MPO) activity determination. In addition, the activity of serum amylase and diamine oxidase (DAO) was determined in each group. RESULTS: The pathohistological damage and MPO of lung, intestine and pancrease, TNF-alpha of BALF, serum amylase and DAO were all increased in AHNP group compared to those in sham operation group (P < 0.05). In AHNP + ligation group, the pathohistological damage and MPO of lung and TNF-alpha of BALF were reduced, but the pathohistological damage and MPO of intestine and pancrease were increased compared with AHNP group (P < 0.05), however the activity of serum amylase and DAO was no changed. In AHNP + drainage group, the pathohistological damage and MPO of lung, intestine and pancrease, TNF-alpha of BALF, serum amylase and DAO were all reduced compared to those in AHNP group (P < 0.05). CONCLUSIONS: Our finding suggest that thoracic duct ligation can reduce neutrophil infiltration and TNF-alpha release and then attenuates lung injury in rats with AHNP, but aggravates the injury of intestine and pancrease. While thoracic duct drainage attenuates the injury of lung, intestine and pancreaseat the same time in rats with AHNP.