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2022 ◽  
Author(s):  
Tian-Ran. Li ◽  
Lan-Ping. Shi ◽  
Yong-Gang. Jia ◽  
Ricky Wing Tong. Lau ◽  
Xia-Zhen. Pan

Abstract Background:Although the circuit condensate, an ideal bacterial reservoir, may flow into the humidifier reservoir (HR), no study has investigated if HR-colonized bacteria colonize other circuit locations with airflow. Therefore, the objective of this study was to explore if bacterial growth in the HR leads to bacterial colonization in the ventilator circuit. Methods: A randomized controlled experiment was performed in a public tertiary hospital in Guangdong Province, China. In vitro mechanical ventilation models (n = 60), divided into sterile water samples (n = 30) and broth samples (n = 30), were established. Sterile water was used for humidification in the ventilation models. The sterile water group contained either Acinetobacter baumannii (n = 15) or Pseudomonas aeruginosa (n = 15) in humidifier water. The broth group was similar to the sterile water group, but brain heart infusion broth was added to the HR. After 24, 72, and 168 h of continuous ventilation, bacteria in the humidifier water and at different circuit locations were sampled and cultured, and the results were analyzed by the Chi-square test. The difference in bacterial concentration at the HR outlet was analyzed by the F test, and P < 0.05 was considered statistically significant.Results:Bacterial culture results of the sterile water samples were negative. Bacteria in the humidifier water continued to proliferate in the broth group, and the bacterial concentration at different times was not significantly different (P > 0.05). With prolonged ventilation, the bacterial concentration at the HR outlet increased (P < 0.05). During continuous ventilation, no bacterial growth occurred at 10 cm from the HR outlet and the Y-piece of the ventilator circuit. The bacterial concentration at the HR outlet was higher in the P. aeruginosa group than in the A. baumannii group (P < 0.05).Conclusions:Sterile water in the HR was not conducive to bacterial growth. Although bacteria grew in the HR and could reach the HR outlet, colonization of other circuit locations was unlikely.


Plant Disease ◽  
2022 ◽  
Author(s):  
Xiang Xie ◽  
Shiqiang Zhang ◽  
Qingjie Yu ◽  
Xinye Li ◽  
Yongsheng Liu ◽  
...  

Camellia oleifera, a major tree species for producing edible oil, is originated in China. Its oil is also called ‘‘eastern olive oil’’ with high economic value due to richness in a variety of healthy fatty acids (Lin et al. 218). However, leaves are susceptible to leaf spot disease (Zhu et al. 2014). In May 2021, we found circular to irregular reddish-brown lesions, 4-11 mm in diameter, near the leaf veins or leaf edges on 30%-50% leaves of 1/3 oil tea trees in a garden of Hefei City, Anhui Province, China (East longitude 117.27, North latitude 31.86) (Figure S1 A). To isolate the causal agents, symptomatic leaves were cut from the junction of diseased and healthy tissues (5X5 mm) and treated with 70 % alcohol for 30 secs and 1 % NaClO for 5 min, and subsequently inoculated onto PDA medium for culture. After 3 days, hyphal tips were transferred to PDA. Eventually, five isolates were obtained. Then the isolates were cultured on PDA at 25°C for 7 days and the mycelia appeared yellow with a white edge and secreted a large amount of orange-red material to the PDA (Figure S1 B and C). Twenty days later, the mycelium appeared reddish-brown, and sub-circular (3-10 mm) raised white or yellow mycelium was commonly seen on the Petri dish, and black particles were occasionally seen. Meanwhile, the colonies on the PDA produced abundant conidia. Microscopy revealed that conidia were globular to pyriform, dark, verrucose, and multicellular with 14.2 to 25.3 μm (=19.34 μm, n = 30) diameter (Figure S1 D). The morphological characteristics of mycelial and conidia from these isolates are similar to that of Epicoccum layuense (Chen et al.2020). To further determine the species classification of the isolates, DNA was extracted from 7-day-old mycelia cultures and the PCR-amplified fragments were sequenced for internal transcribed spacer (ITS), beta-tubulin and 28S large subunit ribosomal RNA (LSU) gene regions ITS1/ITS4, Bt2a/Bt2b and LR0R/LR5, followed by sequencing and molecular phylogenetic analysis of the sequences analysis (White et al. 1990; Glass and Donaldson 1995; Vilgalys and Hester 1990). Sequence analysis revealed that ITS, beta-tubulin, and LSU divided these isolates into two groups. The isolates AAU-NCY1 and AAU-NCY2, representing the first group (AAU-NCY1 and AAU-NCY5) and the second group (AAU-NCY2, AAU-NCY3 and AAU-NCY4), respectively, were used for further studies. Based on BLASTn analysis, the ITS sequences of AAU-NCY1 (MZ477250) and AAU-NCY2 (MZ477251) showed 100 and 99.6% identity with E. layuense accessions MN396393 and KY742108, respectively. And, the beta-tubulin sequences (MZ552310; MZ552311) showed 99.03 and 99.35% identity with E. layuense accessions MN397247 and MN397248, respectively. Consistently, their LSU (MZ477254; MZ477255) showed 99.88 and 99.77% identity with E. layuense accessions MN328724 and MN396395, respectively. Phylogenetic trees were built by maximum likelihood method (1,000 replicates) using MEGA v.6.0 based on the concatenated sequences of ITS, beta-tubulin and LSU (Figure S2). Phylogenetic tree analysis confirmed that AAU-NCY1 and AAU-NCY2 are closely clustered with E. layuense stains (Figure S2). To test the pathogenicity, conidial suspension of AAU-NCY2 (106 spores/mL) was prepared and sterile water was used as the control. Twelve healthy leaves (six for each treatment) on C. oleifera tree were punched with sterile needle (0.8-1mm), the sterile water or spore suspension was added dropwise at the pinhole respectively (Figure S1 E and F). The experiment was repeated three times. By ten-day post inoculation, the leaves infected by the conidia gradually developed reddish-brown necrotic spots that were similar to those observed in the garden, while the control leaves remained asymptomatic (Figure S1 G and H). DNA sequences derived from the strain re-isolated from the infected leaves was identical to that of the original strain. E. layuense has been reported to cause leaf spot on C. sinensis (Chen et al. 2020), and similar pathogenic phenotypes were reported on Weigela florida (Tian et al. 2021) and Prunus x yedoensis Matsumura in Korea ( Han et al. 2021). To our knowledge, this is the first report of E. layuense causing leaf spot on C. oleifera in Hefei, China.


Plant Disease ◽  
2022 ◽  
Author(s):  
Hongyang Wang ◽  
Chuanzhi Kang ◽  
Wang Yue-Feng ◽  
Sheng Wang ◽  
Zhang Yan ◽  
...  

Atractylodes lancea is an important traditional Chinese medicinal plant whose rhizome is used for treating complaints such as rheumatic diseases, digestive disorders, night blindness and influenza. Jiangsu Province is the optimal cultivation location for high-quality A. lancea rhizome. Since June 2019, symptoms of crown rot and leaf rot were observed in about 10-20% of the A. lancea in a plantation (31° 36' 1" N, 119° 6' 40" W) in Lishui, Jiangsu, China. Lesions occurred on the stem near the soil line and on the leaves (Fig. 1A). Disease incidence reached approximately 80-90% by September, 2021 (Fig. 1B) and resulted in severe loss of rhizome and seed yields. For pathogen isolation, ten samples of symptomatic stem segments and ten diseased leaves were collected, surface-sterilized using 5% NaClO solution, rinsed with sterile water, cut into 0.5-2 cm segments, and plated to potato dextrose agar (PDA), and then incubated at 30°C in darkness. Pure cultures of four isolates showing morphological characteristics of Paraphoma spp. were obtained, identified as a single P. chrysanthemicola strain, and named LSL3f2. Newly formed colonies initially consisted of white mycelia; the five-day-old colonies developed a layer of whitish grey mycelia with a grey underside. 20-day-old colonies had white mycelium along the margin and with a faint yellow inner circular part with irregular radial furrows, and the reverse side looking caramel and russet (Fig. 1C). Pycnidia were subglobose (diameter: 5 to 15 μm; Fig. 1D). Unicellular, bicellular or strings of globose or subglobose chlamydospores developed from hyphal cells (Fig. 1E and 1F). The internal transcribed spacer (ITS) region and large subulin-28S of LSL3f2 were cloned using primers ITS1/ITS4 and LR0R/LR7 (Aveskamp et al. 2010, Li et al. 2013), and deposited in GenBank (OK559658 and OK598973, respectively). BLASTn search and phylogenetic analysis showed the highest identity between LSL3f2 and P. chrysanthemicola sequences (Fig. 1G) and confirmed LSL3f2 as P. chrysanthemicola. Koch’s postulates were completed using one-month-old vegetatively propagated A. lancea plantlets growing on autoclaved vermiculite/peat mixture at 26°C with a light/dark cycle of 12/12 hours. Each plantlet was inoculated with 5 ml of conidial suspension in water (1 × 108 cfu/ml) by applying to soil close to the plantlet, with sterile water used as a mock control (n = 10). By 20 days post-inoculation, inoculated plantlets showed a range of disease symptoms consistent to those observed in infested fields (Fig. 1H). Pathogenicity was additionally confirmed using detached leaves inoculated with a colonized agar plug of LSL3f2 or an uninoculated control comparison (diameter = 5 mm) and incubated at 26℃ in the dark. Five to seven days post-inoculation, detached leaves showed leaf rot symptoms including lesions, yellowing and withering consistent with those in infested fields, while control leaves remained healthy (n = 10, Fig. 1I). The pathogen was reisolated from the diseased plantlets and detached leaves, in both cases demonstrating the micromorphological characteristics of LSL3f2. P. chrysanthemicola has been reported to cause leaf and crown rot on other plants such as Tanacetum cinerariifolium (Moslemi et al. 2018), and leaf spot on A. japonicain (Ge et al. 2016). However, this is the first report of P. chrysanthemicola causing crown and leaf rot on A. lancea in China.


2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Ryan Tsz-Hei Tse ◽  
Hongda Zhao ◽  
Christine Yim-Ping Wong ◽  
Angel Wing-Yan Kong ◽  
Ronald Cheong-Kin Chan ◽  
...  

AbstractUrinary bladder cancer is a common cancer worldwide. Currently, the modality of treating and monitoring bladder cancer is wide. Nonetheless, the high recurrence rate of non-muscle-invasive bladder cancer after surgical resection is still unsatisfactory. Hereby, our study demonstrated whether the intra-operative and post-operative environments will affect bladder cancer recurrence utilizing in vitro cell line model. Bladder cancer cell lines were submerged in four different irrigating fluids for assessing their tumorigenic properties. Our results showed that sterile water performed the best in terms of the magnitude of cytotoxicity to cell lines. Besides, we also investigated cytotoxic effects of the four irrigating agents as well as mitomycin C (MMC) in normothermic and hyperthermic conditions. We observed that sterile water and MMC had an increased cytotoxic effect to bladder cancer cell lines in hyperthermic conditions. Altogether, our results could be translated into clinical practice in the future by manipulating the intra-operative and post-operative conditions in order to lower the chance of residual cancer cells reimplant onto the bladder, which in turns, reducing the recurrence rate of bladder cancers.


Plant Disease ◽  
2021 ◽  
Author(s):  
Di Yang ◽  
Chan Juan Du ◽  
Yunfeng Ye ◽  
Lian Fu Pan ◽  
Jin Zhang ◽  
...  

Banana (Musa spp.) is a popular fruit all over the world, and it’s also an important cash crop with a planting area of 358,924 ha in southern China. In July 2020, a peduncle soft rot disease occurred on dwarf banana (Musa sp. cv. Guangfen) in Guigang city (N22°50'29″, E109° 43'34″), Guangxi province, China. More than 20% plants were infected in the banana plantation. The first external sign of the disease appeared on the incisional wound after the flower bud was cut off from the peduncle. The symptom initially appeared as a black lesion on the wound, then extended into the internal tissue of the whole peduncle. In the later stages, the internal tissue became soft and rot, occasionally formed a necrotic cavity, and eventually led to the black rot of the whole peduncle with a foul smell. To isolate the pathogen, the internal lesion tissues of 5 mm × 5 mm were collected between the border of symptomatic and healthy tissue, treated with 75% ethanol for 10 s, and 0.1% HgCl2 for 3 min, then rinsed with sterile water for three times. Sterilized tissue fragments were cut to pieces with sterilized surgical shears and soaked in 5 mL sterile water, then shaken for 10 min in a vortex oscillator. The suspension was diluted 1000 times with sterilized water,then plated on nutrient-agar medium and incubated at 28℃ in darkness for 24 h. Among the 32 isolates, 23 pure bacterial cultures with similar morphology were predominantly obtained from the samples. These bacteria were gram-negative, and their colonies were initially yellowish white with irregular edges and smooth surfaces, then turned to grayish blue after 72 h incubated at 28℃. The representative isolates GZF2-2 and GZF1-8 were selected for further identification. Genomic DNA was isolated from the bacteria and the 16S rDNA was amplified with primers 27F/1492R (Weisburg et al. 1991) and sequenced. The obtained sequences (GenBank Accession No. MZ768922 and OK668082) showed >99% identities to several records of Dickeya fangzhongdai deposited in NCBI GenBank (1400/1404 bps for GZF2-2 to KT992690, 1409/1417 bps for GZF1-8 to MT613398) based on BLAST analysis. In addition, the recA, fusA, gapA, purA, rplB, dnaX genes and the 16S-23S intergenic spacer (IGS) regions of the two isolates were also amplified and sequenced (GenBank Accession Nos. OK634381-OK634382, OK634369- OK634370, OK634373-OK634374, OK634377-OK634378, OK634385-OK634386, OK634365- OK634366 and OK631722-OK631723) as described by Tian et al. (2016). All the DNA sequences matched that of D. fangzhongdai strains JS5T (percent identities>99.06%), PA1 and ECM-1 in GenBank. Neighbor-joining phylogenetic analysis by software MegaX (Kumar et al. 2018) based on the 16S rDNA sequences revealed that the two isolates were in the same clade with reported D. fangzhongdai strains. Multilocus sequence analysis of the other seven regions also showed the two representative isolates were belong to D. fangzhongdai. Therefore, the isolates were identified as D. fangzhongdai. Pathogenicity of isolate GZF2-2 was investigated to demonstrate Koch’s postulate. The end of the banana peduncles of 6 healthy plants were cut off, and 10 mL bacterial suspension (108 CFU/mL) was inoculated to the fresh wound on the plants using sterile brushes. Six control plants were inoculated with sterilized water. All the inoculated peduncles were covered with plastic bags to maintain high humidity. After 28 days, all the peduncles inoculated with strain GZF2-2 showed soft rot symptoms similar to those observed in the field, while the controls remained symptomless. The same bacteria were re-isolated from the symptomatic peduncles and confirmed by sequencing the 16S rDNA. D. fangzhongdai has been reported to cause soft rot on onion (Ma et al. 2020) and bleeding cankers on pear trees (Chen et al. 2020). To the best of our knowledge, this is the first report of D. fangzhongdai causing peduncle soft rot on banana in China.


Author(s):  
M. Schimmoeller ◽  
K. Wulf ◽  
C. Walker ◽  
B. Barrett-Reis ◽  
M. Vurma

BACKGROUND: There is little published data on how to prepare probiotic supplements for enteral delivery in the NICU. The objective of this study was to determine how a three-strain probiotic blend (Bb-02, TH-4 ® and BB-12 ®) would behave when mixed and held for 4 hours with saline water, sterile water, dextrose 5% in water (D5W), 24 kcal preterm formula, and human milk. METHODS: A packet of a three-strain probiotic supplement was mixed with 3 ml of saline water, sterile water, D5W, 24 kcal preterm formula, and human milk (tested at 3 ml and 2 ml). Samples were stored at room temperature for 60 minutes then refrigerated for 180 minutes. Probiotic survival, using quantitative enumeration, and pH were monitored over 4 hours. Samples were passed through a 5 French (Fr) feeding tube at the end of the study to evaluate viscosity. RESULTS: The largest variation in total cell count from 0-time was sterile water with a + 0.26 log(CFU)/mL change at 90 minutes and typical variation is considered±0.50 log units indicating no significant change between samples in 4 hours. Saline water had the lowest final pH at 4.88. All samples easily passed through a 5 Fr feeding tube. CONCLUSION: The study showed minimal change in cell counts across solutions for 4 hours of storage, indicating health care facilities may be able to prepare probiotic supplements with a variety of solutions in pharmacies or milk rooms. This allows greater flexibility for probiotic delivery to preterm infants.


2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Min Li ◽  
Zufeng Chen ◽  
Lin Liu ◽  
Xiaoyun Ma ◽  
Jun Zou

Background. Vitamin C (Vc) has been found to promote corneal wound healing after alkali burns. However, the specific mechanism and functional modes are still unclear. The present study sought to assess the mechanisms of Vc function on corneal alkali burns. Methods. Eighty BALB/c mice were divided into four groups: a normal group without alkali injury (n = 10), an alkali injury group without any treatment (1-day group, n = 10), a Vc group treated with topical 10% Vc (Vc group, n = 30), and a control group treated with topical sterile water (control group, n = 30). Except in the blank control group, the alkali injuries were induced in one eye of each mouse. The mice in the treatment group were given Vc by topical application (q 1 h for 6 days), while those in the control group were given topical sterile water. The clinical evaluations, including corneal fluorescent staining, corneal opacity, and neovascularization, were assessed on days 1, 4, 7, and 10 using slit-lamp microscopy. Ten mice at each time point were sacrificed. The protein expressions in the corneas of p63, PCNA, CK3, MPO, CD31, and α-SMA were detected by immunohistochemistry to examine the corneal epithelial stem cells, corneal epithelium wound healing, corneal stroma inflammation, neovascularization, and fibrosis. Results. The scores of the corneal epithelium defects, corneal neovascularization, and corneal opacities in the Vc group were significantly decreased compared to the control group on day 10. We found that Vc promoted the activation of the corneal epithelial stem cells as shown by a higher number of p63-positive and PCNA-positive cells and an increased CK3 expression when compared with the control group p < 0.001 . The central corneal re-epithelialization was completed by day 10. Moreover, Vc inhibited MPO, CD31, and α-SMA expressions. These results first indicated that the frequent use of topical Vc in the first 6 days of corneal alkali burns alleviated corneal inflammatory cell infiltration, activated corneal epithelial stem cell activity, and reduced corneal neovascularization and fibrosis within 10 days. Conclusions. The study, therefore, showed the therapeutic benefits of Vc on corneal alkali burns and provided new insight into the mechanisms of Vc regulation on corneal wound healing.


2021 ◽  
Vol 8 (12) ◽  
pp. 217
Author(s):  
Riaz Ali ◽  
Justin Bartholomew ◽  
Randolph Coffey ◽  
Caroline Carrico ◽  
Todd Kitten ◽  
...  

Extracted human teeth provide the closest approximation to teeth in situ and play important roles in dental education and materials research. Since extracted teeth are potentially infectious, the Centers for Disease Control recommend their sterilization by autoclaving or disinfection by formalin immersion to ensure safe handling. However, autoclaving is not recommended for teeth with amalgam fillings and formalin is hazardous. The goal of the present study was to investigate the potential of peracetic acid (PA) as an alternative method to achieve reliable disinfection of freshly extracted teeth. A total of 80 extracted teeth were collected for this study. Whole teeth were incubated in one of four solutions for defined periods of time: sterile water (2 weeks), formalin (2 weeks), PA 1000 ppm (2 weeks), and PA 2000 ppm (1 week). After sectioning, the crown and root fragments were transferred into separate tubes containing brain–heart infusion broth and incubated at 37 °C under anaerobic conditions for 72 h. Absence of broth turbidity was used to assess effectiveness of disinfection. No turbidity was observed in any of the formalin-treated or peracetic acid-treated samples, signifying complete disinfection. Our results indicate that PA can effectively disinfect extracted human teeth, providing a reliable alternative to formalin and autoclaving.


Author(s):  
Mayur B. Wanjari ◽  
Deeplata Mendhe ◽  
Pratibha Wankhede

Actilyse can break blood clots that form in the heart, blood arteries, or lungs during a heart attack. This medication is also given to stroke patients to improve recovery and reduce the likelihood of impairment. Recombinant DNA technology was used to create Activase, a tissue plasminogen activator. It is a sterile, purified glycoprotein of 527 amino acids. It is made by combining complementary DNA (cDNA) from a human melanoma cell line with the natural human tissue-type plasminogen activator. After reconstitution with Sterile Water for Injection, USP, Activase is a sterile, white to off-white lyophilized powder for intravenous injection.


Plant Disease ◽  
2021 ◽  
Author(s):  
Yiwen Xu ◽  
Zhenyan Cao ◽  
Yihua Yang ◽  
Xuping Shentu ◽  
Xiaoping Yu

Fritillaria thunbergii Miq. (Zhe beimu), is an oldest known homeopathic traditional folk medicinal plant in Zhejiang Province, China. The bulbs are medicinally important curing cough, inflammation, gastric ulcers, hypertension, diarrhea, and bronchitis (Nile et al. 2021). In April of 2018 and 2019, symptoms of blight on F. thunbergii were observed with an incidence of 20–25% in Cixi city, Zhejiang Province. At the early stage of this disease, the stalk turned brown, then the whole bulbs rotted within a few days. Symptomatic bulbs were cut into small pieces (1.0 cm × 1.0 cm) and disinfected successively by submersion in 75% ethanol for 30 seconds and 1% NaClO for 30 seconds under aseptic conditions. After rinsing with sterile water three times and air drying, segments were placed on potato dextrose agar (PDA). After incubation at 28 ℃ for 7 days in the dark, the hyphae were observed white fluffy, spreading from the middle to the whole plate. Macroconidia were falciform with zero to four septa and (11.0-39.0) × (3.0-5.0) μm in size. Microconidia were fusiform with zero to two septa (4.0-7.0) × (2.6-3.0) μm in size. Based on morphological characteristics of macroconidia, and microconidia, isolates were identified as F. oxysporum (Lombard, L. et al., 2019). The internal transcribed spacer (ITS) region, translation elongation factor (EF-1α), and RNA polymerase second largest subunit (RPB2) gene were amplified and sequenced respectively using ITS1/ITS4, EF1/EF2 and 5f2/7cr primers (O’Donnell et al., 2010). BLASTN analysis of FUSARIUM-ID using ITS (Accession NO.MZ268594), EF-1α (Accession NO.MZ292517) and RPB2(Accession NO.MZ292516) showed 95.2%, 100%, and 99.11% identity to F. oxysporum species complex isolates NRRL43730, NRRL38599 and NRRL38302, respectively. Based on the morphological and molecular characters, the pathogen was identified as F. oxysporum. To verify pathogenicity, ten healthy F. thunbergii plants were used for inoculation tests. One milliliter of a conidial suspension (106 conidia ml-1) was pipetted onto the soil around the base of F. thunbergii plants per pot. Ten plants, which were treated with sterile water, were used as the control. All plants were maintained in a climatic chamber (26 ± 1 ℃, 70–80% relative humidity and a photoperiod of 16:8 [L: D] h). Seven days later, all inoculated plants showed typical symptoms of blight identical to those observed in the fields. Control plants remained symptomless and healthy. The pathogenicity analysis was repeated three times. Pathogens re-isolated from symptomatic plants were identified as F. oxysporum by morphology observation and sequence analysis. To our knowledge, this is the first report of blight caused by F. oxysporum on F. thunbergii in Zhejiang Province, China. Acknowledgments: The author(s) declare no conflict of interest. Funding: This work was supported by Zhejiang Provincial Program for Science and Technology Development (2017C32006, 2018C02030) and the Student Science and Technology Innovation Project of China Jiliang University (2021YW95). References: Nile et al. 2021.J. Food and chemical toxicology, 153:112289. Hami, A. et al., 2021. J. Scientific Reports. 3610.11.1. Lombard, L. et al., 2019. PERSOONIA, 43:1-47. O’Donnell, K., et al. 2010. J. Clin. Microbiol. 48: 3708-3718.


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