Ml Of Sterile Water Research Articles

Nintedanib alleviates polypropylene mesh-induced adhesion in rabbit ventral herniorrhaphy

Abstract BACKGROUND: Nintedanib (previously known as BBIF 1120) was approved by the United States Food and Drug Administration and European Medical Agency to treat idiopathic lung fibrosis in the year 2014 and 2015, respectively. It is now gaining interest for its anti-fibrotic activity in other organs and disease conditions. Although a surgical mesh is used as a mainstay therapy for herniorrhaphy, postoperative peritoneal adhesion with a polypropylene mesh is a significant drawback. This study aims to assess the efficacy of nintedanib in preventing postoperative adhesion incited with a polypropylene mesh in a rabbit model of ventral hernia. MATERIALS AND METHODS: Ventral hernia was induced surgically in ten adult healthy New Zealand White rabbits of either sex. Hernioplasty was performed with a polypropylene mesh, and the rabbits were randomly allocated into two groups to receive either oral 1 ml sterile water or 100 mg nintedanib in 1 ml of sterile water for 7 days. The adhesion of the implanted mesh with the intra-abdominal organs was assessed clinically, by histological and ultrastructural studies with scanning electron microscopy (SEM) at 30 days postoperatively. RESULTS: All rabbits were clinically healthy for 30 days post-surgery with no complication at the site of surgery. The incidence of peritoneal adhesion and tenacity was less in the nintedanib group versus the control group (P = 0.00105). Histopathological and SEM evaluations also indicated less fibrosis and adhesion in the nintedanib-treated group versus control. CONCLUSION: Our results show successful prevention of mesh-associated adhesion with nintedanib, but further studies on the mechanistic pathway, pharmacokinetics, dose standardization, and evaluation of systemic side effects are warranted.

Midface Projection Using Biostimulatory Poly-l-Lactic Acid Injectable Implant: A Subgroup Analysis of the Cheek Wrinkle Trial.

Correction of cheek wrinkles using poly-l-lactic acid (PLLA-SCA) was demonstrated in a 12-month study. This analysis assessed change from baseline in lifting effect of PLLA-SCA using a 3D camera to provide additional quantified data. Subjects received PLLA-SCA (reconstituted in 8 mL of sterile water + 1 mL of 2% lidocaine) in both cheeks or no treatment (control). Assessments included the Galderma Cheek Wrinkle Scale (GCWS), aesthetic improvement, and 3D photography. In subjects with severe GCWS at baseline, Canfield software analyzed 3D images for change from baseline in midface volume projection and volume change of both cheeks at Month 9. The primary study showed a statistically significant higher at-rest GCWS responder rate at Month 12 for PLLA-SCA, 71.6%, versus control, 26.1% (p < .0001). In this Month 9 analysis, mean midface volume projection demonstrated a positive volume shift for PLLA-SCA (left: 0.45 mm; right: 0.34 mm; n = 46) versus control (left: 0.25 mm; right: -0.01 mm; n = 21). Midface volume and max projection changes for PLLA-SCA were +4.88 mL/+1.62 mm (left), +2.84 mL/+1.12 mm (right) versus +0.26 mL/+0.34 mm (left), +0.68 mL/+0.37 mm (right) for control. Poly-l-lactic acid-treatment of subjects with severe GCWS had a clinically meaningful lifting effect in both cheeks (positive volume shift, positive volume change, and max projection) in the midface. NCT04124692.

Citric Acid by-Product Fermentation by Bacillus subtilis I9: A Promising Path to Sustainable Animal Feed

Citric acid by-products in animal feed pose a sustainability challenge. Bacillus species are commonly used for fermenting and improving the nutritional quality of feedstuffs or by-products. An experiment was conducted to enhance the nutritional value of citric acid by-products through fermentation with Bacillus subtilis I9 for animal feed. The experiment was carried out in 500 mL Erlenmeyer flasks with 50 g of substrate and 200 mL of sterile water. Groups were either uninoculated or inoculated with B. subtilis I9 at 107 CFU/mL. Incubation occurred at 37 °C with automatic shaking at 150 rpm under aerobic conditions for 0, 24, 48, 72, and 96 h. Inoculation with B. subtilis I9 significantly increased Bacillus density to 9.3 log CFU/mL at 24 h (p &lt; 0.05). CMCase activity gradually increased, reaching a maximum of 9.77 U/mL at 72 h. After 96 h of fermentation with inoculated B. subtilis I9, the citric acid by-product exhibited a significant decrease (p &lt; 0.05) in crude fiber by 10.86%, hemicellulose by 20.23%, and cellulose by 5.98%, but an increase in crude protein by 21.89%. Gross energy decreased by 4% after inoculation with B. subtilis in comparison to the uninoculated control (p &lt; 0.05). Additionally, the non-starch polysaccharide (NSP) degradation due to inoculation with B. subtilis I9 significantly reduced (p &lt; 0.05) NSP by 24.37%, while galactose, glucose, and uronic acid decreased by 22.53%, 32.21%, and 18.11%, respectively. Amino acid profile content increased significantly by more than 12% (p &lt; 0.05), including indispensable amino acids such as histidine, isoleucine, lysine, methionine, phenylalanine, tryptophan, and valine and dispensable amino acids like alanine, aspartic acid, glutamic acid, glutamine, glycine, proline, and tyrosine. Furthermore, citric acid by-products inoculated with B. subtilis I9 exhibited changes in the cell wall structure under scanning electron microscopy, including fragmentation and cracking. These results suggest that fermenting citric acid by-products with B. subtilis I9 effectively reduces dietary fiber content and improves the nutritional characteristics of citric acid by-products for use in animal feed.

Effect of intrafollicular administration of PGE2 or PGF2α in early estrus on ovulation, hemorrhagic anovulatory follicles formation, progesterone secretion and pregnancy outcome in the mare

This experiment was performed to evaluate whether intrafollicular treatment of PGE2 or PGF2α administered in early estrus would induce normal ovulation, progesterone production (Experiment 1) and pregnancy (Experiment 2). In Experiment 1, mares in estrus after 2 days of endometrial edema were injected in all largest dominant follicles (28–35 mm in diameter) with 0.5 mL of sterile water containing 500 μg PGE2 (n = 6), 125 μg PGF2α (n = 6) or placebo (n = 7) (Hour 0). Ultrasound examinations were performed daily, until ovulation or anovulation was detected, and daily blood samples were taken for 8 days. In Experiment 2, mares with a dominant follicle ≥35 mm after at least three days of slight-to-moderate endometrial edema, were injected with 500 μg PGE2 diluted in 0.5 mL of sterile water for injection in the follicle (PGE2 group; n = 9 mares and 11 dominant follicles). No puncture was performed in the control group (n = 9 mares and 11 dominant follicles). Mares from both groups were inseminated. In Experiment 1, all mares (6/6) in the PGE2 group ovulated within 24 h of treatment. The mean interval from intrafollicular injection to ovulation was shorter (P < 0.001) in PGE2 mares (24 ± 0 h) than in control mares (77 ± 9 h). Mares from the PGF2α group developed hemorrhagic anovulatory follicles (HAF) more often (7/7) than control mares (2/7); P < 0.05). The progesterone concentration in mares from the PGF2α group was lower (P < 0.004) than control mares in the early post-ovulatory period. The first significant increase in post-ovulatory progesterone concentration occurred earlier (P < 0.05) in mares from the control group than in mares from the PGF2α and PGE2 groups. In Experiment 2, more mares from the control group (7/9, 78 %) became pregnant than from the PGE2 group (2/9, 22 %) (P = 0.015). In conclusion, PGE2 alone induced follicle collapse in all treated mares within 24 h of administrations, while PGF2α blocked ovulation and induced formation of HAFs. However, the post-ovulatory rise in progesterone production was delayed and the fertility reduced in mares with ovulation induced by PGE2 compared to control mares.

Assessment of chemical contamination by cancer drugs during use of the RIVATM compounding robot: A pilot study.

Many hospitals are now investing in robotic compounding system for the preparation of cytotoxic agents. The objective of the present study was to describe contamination by cytotoxics inside and outside the RIVATM robot (ARxIUM, Winnipeg, Canada). We applied a risk analysis to determine which locations inside and outside the compounding robot should be monitored. Samples were collected by swabbing with a wet swab (using 0.1 mL of sterile water) before the robots was cleaned. Ten cytotoxics compounded with the robot were screened for using LC-MS/MS. We determined the percentage contamination rates inside (CRin) and outside (CRout) the robot and the amounts of each contaminant (in ng/cm²). If a sample was found to be positive, a corrective action was implemented. Our risk analysis highlighted 10 locations inside the robot and 7 outside. Ten sampling campaigns (10 samples per campaign) were performed. The mean CRin (40%) was significantly higher than the mean CRout (2%; p < 10-4). Gemcitabine and cyclophosphamide were the main contaminants. After the implementation of corrective measures (such as daily cleaning with SDS/isopropyl alcohol), the CRin fell from 60% to 10%. The frequency of contamination was lower for robotic compounding than for manual compounding in an isolator. However, robotic compounding tended to generated larger mean amounts of contaminant; this was related to incidents such as splashing when syringes were disposed of after the compounding. The implementation of corrective actions effectively reduced the CRs. Further longer-term studies are required to confirm these results.

First report of two Pantoea species causing bacterial leaf blight on wheat in the United States.

Wheat (Triticum aestivum) is an important crop worldwide, contributing to about one third of the global caloric intake. In June 2021, leaves with bacterial blight symptoms, including yellow and necrotic lesions running parallel to veins, were found in several fields across five counties in eastern Colorado (Weld, Morgan, Sedgwick, Baca, and Kit Carson). Plants exhibiting these symptoms were scattered throughout fields, but symptoms appeared consistent across counties. To determine the causal agent and complete Koch's postulates, a 1 cm2 symptomatic leaf area was excised and macerated in 0.5 mL of sterilized water from four field samples. The lysate was spread on yeast extract dextrose calcium carbonate medium (YDC agar, 1% yeast extract, 2% dextrose, 2% calcium carbonate, 1.5% agar) to isolate bacteria. Single colonies of yellow, mucoid morphology were selected and streaked on new YDC plates. Isolate genomic DNA was extracted (Zymo Research Quick-DNA Fungal/Bacterial Miniprep Kit, #D6005), and ~30 ng of gDNA was used to amplify the 16S rRNA, gyrB, and rpoB genes of all four isolates (Barret et al., 2015; Delétoile et al., 2009; Krawczyk et al., 2020; Ogier et al., 2019). Amplified PCR products were cleaned (Zymo DNA Clean & Concentrator kit, #D4033) and Sanger sequenced, and all sequences have been deposited in NCBI (16S rRNA: OR707336, OR707337, OR707338, OR707339), (gyrB: PP407951, PP407952, PP407953, PP407954), (rpoB: PP407955, PP407956, PP407957, PP407958). A BLAST search against whole genomes identified one isolate from Kit Carson county (CO314) and two isolates from Baca county (CO316 and CO317) as Pantoea agglomerans with 100% identity for the 16S rRNA, gyrB, and rpoB genes, and one isolate from Weld county (CO315) was 100% identical to Pantoea allii for all three genes. To complete Koch's postulates and confirm Pantoea sp. as the causal disease agents, isolates were grown as lawns on DifcoTM Nutrient Agar (NA) medium (48h, 28℃), suspended in 10 mM MgCl2 using a final optical density of 0.1 (~109 colony forming units per milliliter (CFU/mL)), and syringe-infiltrated into the entire leaf area of 10-day-old wheat seedling leaves (var. Hatcher). Treatments of 10mM MgCl2 and a field isolate that does not cause symptoms, identified as Pseudomonas synxantha by 16S rRNA and gyrB sequencing, were negative controls. Inoculated wheat plants were transferred to a growth chamber (22℃, 90% relative humidity). Symptoms developed 14 days post inoculation (dpi), with the most severe appearing 21 dpi. Each of the four Pantoea isolates were re-isolated from symptomatic leaves by grinding them in a Tissue Lyser II (Qiagen) with two metal beads and diluting with 0.4 mL of sterile water. A 20 µL sample of each isolate was plated on NA (24h, 28℃). The colonies appeared phenotypically identical to the original isolates, and Sanger sequencing confirmed the identities of the isolates. To our knowledge, this is the first report of P. agglomerans causing disease in wheat in the United States, and the first report of P. allii as a wheat disease-causing agent. This report is consistent with previous communications showing P. agglomerans causing wheat disease in China (Gao et al., 2023), and P. ananatis in Poland (Krawczyk et al., 2020). The growing numbers of reports of Pantoea spp. as pathogens in recent years suggests increasing novel disease emergence on cereals worldwide.

Isolation, Characterization, and Selection of Bacillus sp. from Shallot Rhizosphere that Inhibits Fusarium oxysporum Growth

Bacillus sp. is a Plant Growth Promoting Rhizobacteria (PGPR) species that lives in the rhizosphere. This bacterium can produce antifungal compounds that suppress pathogenic fungi, such as Fusarium oxysporum. The aim of this research was isolate, characterize, and obtain Bacillus sp. that can inhibit F. oxysporum which causes twisted disease in shallots. Isolation was done by sampling 10 g of soil from shallot roots, placing it in 9 mL of sterile water, and carrying out a series of dilutions. The first dilution was incubated at 80 °C for 20 minutes to obtain Bacillus sp. on the selective medium HiChrome. Colony morphology, shape, cell color, Gram staining, catalase and endospore staining were observed from obtained isolates. An antagonist test was done to determine the inhibitory effects of isolates against the pathogenic fungus F. oxysporum. Four isolates of Bacillus sp. were obtained with irregular colony shape, dull white color without muccus, edges varying in shape between lobate and undulate, and raised elevation. The results of the antagonist test showed that the four isolates had the ability to inhibit the growth of the pathogenic fungus F. oxysporum with the greatest inhibition shown by isolate BM1 at 30.12%, with an antibiosis mechanism and hyphae swelling.

Resistance of Red Maple and Freeman Maple Cultivars to Phytopythium vexans, a Causal Agent of Root and Crown Rot Disease.

Phytopythium root rot caused by Phytopythium vexans is an emerging threat to red maple and Freeman maple production that seriously impacts plant growth, aesthetic, and economic values. This study reports on the resistance of red maple and Freeman maple cultivars against root rot disease caused by P. vexans. Rooted cuttings were received from a commercial nursery and planted in 3-gallon containers. For each cultivar, six plants were inoculated by drenching 300 ml/plant of P. vexans suspension, prepared by blending two plates of 10-day-old P. vexans culture/liter sterile water. An equal number of plants remained noninoculated and were drenched with 300 ml of sterile water. Two trials were conducted for 4 months in the greenhouse during the summer of 2023. Plants were evaluated for growth, physiology, Phytopythium root rot severity (0 to 100% of roots affected), and pathogen reisolation frequency. Out of seven cultivars, Somerset had the lowest Phytopythium root rot severity and pathogen reisolation frequency. Cultivars Autumn Blaze, Brandywine, and October Glory were highly susceptible to P. vexans, whereas Sun Valley, Summer Red, and Celebration were found to have a partial resistance to P. vexans. Cultivars Autumn Blaze, Brandywine, and October Glory had significantly lower chlorophyll content, net photosynthesis, and stomatal conductance compared with the other three cultivars under pathogen inoculation. Phytopythium root rot reduced plant height, width, total plant, and root fresh weight. The disease severity was negatively correlated with width, chlorophyll content, net photosynthesis, and stomatal conductance. These results can aid growers and landscapers in developing effective P. vexans management strategies.

The effect of octreotide on the duration of postmastectomy wound drainage: A randomized controlled trial

Background: Following mastectomy, prolonged wound drainage delays adjuvant therapy and also prolongs hospitalization in an otherwise fit patient. This study determined the effect of octreotide on the duration of postmastectomy wound drainage. Methods: Patients scheduled for modified radical mastectomy were randomized into a treatment group that received 100 µg of octreotide and a control group that received 1 mL of sterile water intravenously 8 hourly for 5 days from the first postoperative day. Sociodemographic data, time to drain removal, total drain volume, and incidence of seroma were obtained. The groups were compared for the time to drain removal, total drain output, average drain output per day, and incidence of seroma. Statistical significance was set at P &lt; 0.05. Results: Forty-one female breast cancer patients were recruited. The mean age was 53.1 ± 13.0 years. The most common histologic variant was invasive carcinoma of no special type (n = 27; 65.9%). Although not statistically significant, patients in the intervention group had a lower median time to drain removal of 11 days (interquartile range [IQR] = 3) compared with 13 days (IQR = 3) in the control group (P = 0.106) and similarly had a lower median total drain output compared with controls (892.0 mL vs. 1182.8 mL; P = 0.095). Both groups were comparable in terms of postoperative complications and these occurred in 13 patients (31.7%), the most common being seroma (n = 11; 50.0%). Conclusion: Octreotide had no statistically significant effect on either the total drain output or the duration of postmastectomy wound drainage among breast cancer patients.

First Report of Bulb Rot Disease of Banana Caused by Klebsiella variicola in China.

Banana (Musa spp.) is an economically important fruit and food crop globally as well as in China. In March 2023, a bulb rot disease was observed on more than 20% of cultivated dwarf bananas in a plantation in Wuming County of Guangxi Province, a major hub of banana production in China. Infected plants showed crackles at the basal part of stem and were relatively dwarf, while yellowing of the leaves was not observed. When the rhizomes were cut open, water-soaked lesions with a yellow or black margin can be seen in the bulb. In severe infections, the internal tissue became dry or wet rot, and there was typical dark-brown cavity formation in the bulb. The rot was limited to the bulb. To isolate the causal agent, dissected diseased tissues (5×5 mm) were surface sterilized with 75% ethanol (30 s) and 2% NaClO (3 min), followed by three rinses with sterile water. The sterilized sections were soaked in 2 mL of sterile water and shaken for 5 min in a vortex oscillator. The suspension was streaked on Luria-Bertani (LB) agar medium, and incubated at 28℃ for 24 h. Single colonies were re-streaked three times to obtain purified isolation. Twelve pure bacterial cultures with similar morphology were isolated from three plants taken from the field. The bacterial colonies were yellowish white, mucoid, round, and raised with translucent surfaces on the LB agar plate. Three strains Gxkv1, Gxkv2 and Gxkv3 were selected for further analyses. The 16S rDNA gene (GenBank Accession OR461756, PP094726 and PP109349) were amplified using primer pair 27F/1492R (Frank et al. 2008). Comparing 16S sequences against GenBank showed 99.86%-100% sequence identity to Klebsiella variicola strain (MZ475068) for the three isolates Gxkv1 (1,398/1,398 bp), Gxkv2 (1,398/1,396 bp) and Gxkv3 (1,398/1,398 bp). A multilocus phylogenetic analysis was conducted by neighbor-joining method (1,000 bootstrap values) based on three housekeeping gene sequences of gyrA (GenBank Accession No. OR515493, PP105747, PP105748), rpoB (OR515494, PP105751, PP105752 ) and infB (OR515495, PP105749, PP105750) genes which were amplified by gyrA-A/gyrA-C, CM31b/CM7 and infB867F/infB1819R primer sets, respectively (Rosenblueth et al. 2004). The results of phylogenetic analysis showed the three strains belong to the K. variicola clade. A pathogenicity test was conducted on six healthy 3-month-old dwarf banana plants by spraying 10 mL of bacterial suspensions of Gxkv1 (108 CFU/mL) into the rhizome which wounded with a sterilized needle; another six healthy control plants were sprayed with 10 mL of sterile water. Following inoculation, the plants were placed in a greenhouse at 28-32°C. After 30 days, all inoculated plants showed symptoms similar to those observed in the field, while the control plants remained healthy. Bacteria were successfully reisolated from the symptomatic tissues and identified to be K. variicola by PCR mentioned above. K. variicola has been reported to cause rhizome rot of banana in India (Loganathan et al. 2021), and to cause plantain soft rot in Haiti (Fulton et al. 2021). Besides, previous reports from China only showed K. variicola causing banana sheath rot (Fan et al. 2015, Sun et al. 2023). To our knowledge, this is the first report of bulb rot disease of banana caused by K. variicola in Guangxi Province, China. This finding will provide important information for studying the epidemiology and management of this pathogen.

A comparative evaluation of the effect of addition of 8 mg dexamethasone to 2% lignocaine with adrenaline in mandibular third molar surgery: a split mouth randomised double blind study.

Impacted lower third molar surgeries involve trauma in a highly vascularized zone with loose connective tissue leading to inflammatory sequelae including postoperative pain, swelling, trismus and generalised oral dysfunction during the post-operative phase. In minor oral surgical procedures, an all-inclusive method to protract anaesthesia and reduce the inevitable post-operative sequelae is yet to be explored substantially. To evaluate the efficacy of dexamethasone added to local anaesthetics in extending the depth and duration of anaesthesia and decreasing the postoperative complications after surgical removal of impacted third molars. A controlled, randomized, split-mouth, double-blind prospective study involving lower third molar surgery was performed in 35 patients wherein the test group (Group I) received 8 mg dexamethasone added to 2 ml of 2% lignocaine with epinephrine and the control group (Group II) received 2 ml of sterile water added to 2 ml of 2% lignocaine with epinephrine. Onset and duration of anaesthesia were evaluated; followed by evaluation of pain, swelling and trismus for 7 days post-surgery, using independent t-test and ANOVA for repeated measures. Test group had a faster onset of anaesthesia by 69 s and a lengthier duration of 128.4 min (p < 0.001). Pain scores (Visual Analogue Scale) in the first 24 h were 4.9 and 7.5 in the test and control group respectively (p < 0.001). The average dosing of analgesics until postoperative day 7 in the test and control group were 12.6 and 18.4 respectively (p < 0.001). The swelling was significantly lesser in the test group, in addition, trismus was significantly lesser by 1 cm on postoperative days 1 and 2 and 0.2 cm on day 7. The addition of dexamethasone to lignocaine in the nerve block reduces the time of onset and significantly prolongs the duration of anaesthesia with decreased pain, swelling and trismus. Steroids mixed directly with the local anaesthetic agent can minimise the post-operative sequelae associated with third molar surgery with a single needle prick.

First report of bacterial leaf spot on Hami melon caused by Pseudomonas aeruginosa in China.

Hami melon (Cucumis melon var. saccharinus) is an economically important crop all over the world. It is being extensively planted in greenhouse in the southwest part of Hainan province, China. A new bacterial leaf spot was observed in a 20 hm2 Hami melon plantation in Huangliu town, Ledong county, Hainan province, in January 2022. The incidence of the disease was approximately 5%. Symptoms were irregularly shaped, brown lesions with yellow haloes on mature leaves, and first appeared as small, dark-green, water-soaking spots. Specimens from the lesion margin were disinfected by submersion in 0.1% mercuric chloride for 1 min, then soaked with 75% alcohol for 30 s, and rinsed with sterilized distilled water. The tissues were then ground in 2 ml of sterile water and allowed to stand for 5min. The supernatant was streaked onto nutrient agar (NA) and incubated for 48h at 30°C. Colonies were round, smooth, colorless, nearly transparent, bead-shaped at first, and then became lightly blue. After being cultured for days on NA at 30℃, the bacteria can turn the media brown. Yellow green pigments (pyoverdin) that fluoresce under ultraviolet light could be produced by the isolates in the Luria Broth. The bacteria were gram-negative, rod shaped with a single polar flagellum, 0.4 to 1.1 × 1.4 to 3.4 µm. Its physiological and biochemical characteristics were as follows: positive for the oxidase, aerobic, arginine dihydrolase, gelatin liquefaction, denitrification, lipase, growth at 41℃, utilization of mannitol, and production of pyocyanin tests; negative for the hydrolysis of starch, levan formation, lecithinase, growth at 4℃, growth in media supplemented with 8.5% NaCl, and utilization of maltose, xylose, and ethylene glycol tests. The 16S rRNA (1,437 bp), gyrB (1,181 bp), and rpoB genes (1,510 bp) were amplified with 27F/1492R (Zhang et al. 2016）, UP-1s/UP-2sr（Hannula M，2007）, and rpoB-F/rpoB-R (Ogier, JC. et al., 2019) primer sets respectively. One of the 5 isolates collected was sequenced. A BLASTn search of GenBank revealed that the sequence of 16S rRNA gene (OQ918303) had 99.7% identity and 98% coverage comparing with the best hit Pseudomonas aeruginosa strain DSM 50071(NR_117678.1), and both gyrB (OR261077) and rpoB (OR261078) had 99.9% identity and over 98% coverage comparing with P. aeruginosa E90 (CP044006.1). A pathogenicity test was conducted by spraying a suspension of the bacteria (108 CFU/mL) onto 10 Hami melon seedlings with two true leaves. Controls were inoculated with sterile water. All inoculated plants were maintained at 28℃ with 80 to 85% relative humidity in a greenhouse. Dark-green, water soaking spots appeared on the cotyledon and stems of treated seedlings 3-5 days after inoculation, and dark green lesions with halos were observed on the true leaves at the same time. Symptoms did not occur on the control plants. The bacteria which were re-isolated from the inoculated plants were confirmed as P. aeruginosa based on the 16S rRNA gene sequence. The bacterium was not isolated from control plants. P. aeruginosa has been reported to cause disease on a variety of plants including tomato (Zhang et al., 2021), poplar (Liu, et al., 2019), ginseng (Gao et al., 2014), tinda (Mondal et al., 2012), onion (Abd-Alla et al., 2011), tobacco (Yu et al., 2008) and sweet basil (Walker et al., 2004). As far as we know, this is the first report of P. aeruginosa causing leaf spot on Hami melon in China.. This report will contribute to the recognition and diagnosis of the new disease for the Hami melon growers.

First Report of Epicoccum nigrum Causing Brown Leaf Spot of Sweet Cherry (Prunus avium) in China

Sweet cherry (Prunus avium L.) has become an important economic fruit in China, mainly produced in Shandong Province. In recent years, the planting area of Aba Prefecture in Sichuan Province has increased. In June 2022, sweet cherry brown leaf spot was found in a cherry plantation (100ha) in Wenchuan County (30°54′50.21″N, 103°24′49.10″E), with an incidence of 50 - 70%. The symptoms appeared as brown circular spots on the leaf, gradually expanding until multiple lesions coalesced to form large irregular brown spots; eventually entire leaves were killed. To isolate the causal pathogens, 10 diseased trees were randomly selected from an orchard, one diseased leaf was taken from each tree, and samples (4×4 mm2) were cut from the border between diseased and healthy tissues of 10 diseased leaves, surface sterilized with 75% ethanol for 30 sec, washed three times with sterilized water, dried on sterilized filter paper and placed on potato dextrose agar (PDA). After 5d at 25℃, five morphologically similar colonies were obtained, colony appears yellow fluffy and released a large amount of red-orangepigment. Microscopy revealed circular to ovoid, verrucose, and multicellular conidia measuring 20×25 μm diameter (n = 30) were produced on the mycelia. The morphological characteristics were consistent with the description of Epicoccum nigrum (Lima et al 2011). To further identify the strains, the internal transcribed spacer (ITS), β-tubulin, and RNA polymerase second largest subunit (RPB2) gene regions were amplified with ITS1/ITS4 , Bt2a/Bt2b, and 5f2/7cr (White et al. 1990; Glass and Donaldson 1995; Sung et al. 2007), respectively. BLAST analysis revealed that the ITS, β-tubulin, and RPB2 sequences were 99.2%, 100% and 99.6% homologous, with those of E. nigrum (KU204750.1, OL782123.1, and MW602294.1), respectively. The sequences of the five isolates were identical; and those of representative strain TY3 were deposited in GenBank (ITS, OP410968; β-tubulin, OR502448; RPB2, OP484927). Maximum likelihood phylogenetic analyses were performed for the combined data set with ITS , β-tubulin and RPB2 using MEGA6 under the Tamura–Nei model (Tamura et al. 1993). Isolate TY3 clustered with E. nigrum type strain CBS 505.85. The pathogenicity of TY3 was tested on 10 sweet cherry trees aged 3 years (there were about 50 leaves per plant). Five plants were sprayed with 50 mL of spore suspension (1×105 spores/mL), while the controls (Five plants) were sprayed with 50 mL of sterile water. All plants were in closed plastic bags to maintain high humidity, placed in a greenhouse, and incubated at 25℃with a 12-h photoperiod. Twelve days after inoculation, 35% of the inoculated leaves showed lesions; that were consistent with those observed in the field, and the control group was asymptomatic. To confirm Koch´s postulates, two isolates were taken from the margins of leaf lesions and both were confirmed to be E. nigrum based on morphological observations and molecular identification using ITS β-tubulin, and RPB2 sequences. This is the first report of brown leaf spot caused by E. nigrum on P. avium in China. This discovery needs to be considered in developing and implementing disease management programs in sweet cherry production.

OGC O09 Fluorescence lymphography using indocyanine green in the prevention of chyle leakage after minimally invasive esophagectomy

Abstract Background For oncological reasons, the thoracic duct is resected during esophagectomy for cancer. Its anatomy and vulnerability render it prone to injury and mainly side branches and collaterals are often hard to visualize. Chyle leakage is one of the most challenging complications in esophagectomy, with an incidence of 14-21%. The clinical implications for patients are prolonged thoracic drainage and hospital admission, additional reinterventions, unpleasant dietary restrictions, and decreased long-term survival. Fluorescence lymphography using indocyanine green (ICG) can accurately visualize the thoracic duct and collaterals, which may possibly reduce the incidence of chyle leakage and intra-operative detected chyle leakage can be treated directly. Methods A prospective observational cohort study will be performed. Patients aged over 18 years with resectable (cT1-4a, N0-3, M0) esophageal carcinoma undergoing a minimally invasive esophagectomy will be included. The primary endpoint of the study is postoperative chyle leakage. Secondary endpoints are overall morbidity and mortality, nature of dietary restrictions, reinterventions, hospital costs, 2 year survival rate and quality of life. Twenty-five milligram of ICG is dissolved in 10 ml of sterile water, obtaining a solution of 2.5 mg ICG per mL. In the McKeown esophagectomy, before the thoracic phase, the surgeon administers a bilateral bolus of 2 mL ultrasound-guided into inguinal lymph nodes. In an Ivor Lewis esophagectomy, 2mL of reconstituted ICG will be injected into the small bowel mesenteric root just under the peritoneum of the mesentery, at the level where the jejunostomy is placed. Additionally, 50 mL of cream is enterally administered through the just established feeding jejunostomy. Subsequently, the operating area is inspected for chyle leakage with fluorescence lymphography after thoracic duct ligation and just before terminating the procedure. Results We aim to include 50 patients to assess the feasibility of fluorescent lymphography using ICG and enteral cream to prevent chyle leakage after minimally esophagectomy. Inclusion is ongoing. At this time 39 of 50 patients are included. Results will be expected before the congress. Conclusions It is hypothesized that as a result of this technique the patients will benefit from lower incidence of postoperative chyle leakage, shorter thoracic drainage, reduced hospital admission, reduced dietary restriction, less re-interventions, and improved overall survival.

First Report of Diaporthe fruit rot and stem canker of hydroponic cucumber caused by Diaporthe sp. in the United States.

Multiple Diaporthe spp. cause root and fruit rots or stem lesions on Cucumis spp.: D. cucurbitae, D. melonis, D. longicolla (syn. D. eres), D. pterocarpi, D. sclerotioides, D. sojae, and D. ueckerae (Broge et al. 2020; Fukada et al. 2018; Udayanga et al. 2012, 2015). From May-August 2021, cucumbers (Cucumis sativus) 'Katrina' and 'Alcazar' were grown in a 24-plant, commercial Bato bucket system with rockwool blocks on a perlite substrate in a research greenhouse in Wooster, Ohio. At maturity, plants collapsed rapidly from stem lesions without foliar chlorosis (25% of 'Katrina' and 17% of 'Alcazar'). Lesions were 7.5 to 15 cm in length, tan to golden-brown with black pycnidia and located 5 to 15 cm above the crown. Stems shredded easily with vascular discoloration around the lesion. Two identical fungal strains were isolated on ½ acidified potato dextrose agar (APDA) following surface disinfestation with 0.6% sodium hypochlorite for 30 s and sterile water rinse. Fungal cultures were floccose, white to tan mycelia with pycnidia. Oblong, elliptical, biguttulate, aseptate alpha conidia were observed with mean dimensions: 8.0 µm (5.2-9.8 µm) by 3.1 µm (2.5-3.8 µm) on ½ APDA and 9.8 µm (6.6-12.4 µm) by 3.0 µm (1.9-5.3 µm) on petioles. On prune extract agar, beta conidia mean dimensions were: 19.7 µm (12.0-27.7 µm) by 1.2 µm (0.8-1.8 µm). Fungal DNA was amplified and sequenced bidirectionally with ITS (ITS4/ITS5), CAL (CAL228F/737R), HIS (CYLH3F/H3-1B), TEF1 (EF1-728f/EF1-986R), and TUB2 (Bt1a/Bt1b) primers (Carbone and Kohn 1999; Glass and Donaldson 1995) (GenBank: OP265712-13, OP288460-65, OQ418506-07). Based on a maximum likelihood phylogenetic tree of concatenated genes, this novel Diaporthe sp., most closely related to D. stewartii, has not been reported on Cucumis spp. Strains were deposited in the USDA-ARS Culture Collection (NRRL# 64461-62). Koch's postulates were conducted in a greenhouse with mean day temperature of 25°C and 12 hr supplemental lighting. One-month old cucumbers 'Katrina,' grown in rockwool cubes (5 plants per isolate) and potting mix (6 plants per isolate), were inoculated with a one-week-old culture of either strain. The second true leaf was cut and a pipette tip containing an inoculated plug of ½ APDA was placed on the remaining petiole (Mathew et al. 2018). Non-inoculated ½ APDA was used for controls. Plants were tarped for 24 hours to increase humidity and pipette tips removed after one week. After two weeks, petioles were shrunken, tan to golden brown with pycnidia. After 3-4 weeks, stem lesions matching those above were observed on inoculated plants, and plants collapsed. For fruit rot, three Beit Alpha cucumbers were rinsed with tap water, dried, a 5 mm plug was removed from the fruit and replaced with a 5 mm plug of one-week-old fungus on ½ APDA. After 3 days, fruits were water soaked and soft. For root rot, two plates of one-week-old cultures were macerated in 500 mL of sterile water and mixed with 1500 mL of vermiculite. Two seeds of cucumber 'Katrina' were planted into three reps of each isolate and control. All control seeds germinated, but all inoculated seeds experienced pre- or post-emergence damping off. No symptoms were ever observed on any controls. Fungi were isolated from all inoculated tissues as described above. Based on morphology, Diaporthe sp. was isolated from all inoculated plants but never from controls. This Diaporthe sp. may be a new constraint to hydroponic cucumber production, but incidence needs to be determined globally.

Effect of Clomiphene Citrate on Estrus Cycle and The Ova Shed in the Ovary of Female Sprague Dawley Rats

Purpose: Clomiphene citrate (CC) is used in the management of infertile females. Estrus Cycle indices are good parameters for evaluating reproductive changes, while actual ova shed represents evidence of ovulation. This study is aimed at investigating how different doses of CC can affect the Estrus Cycle (EC), Estrus Cycle Ratio, (ECR), and ova shed (OS) in adult female rats. &#x0D; Methodology: Twenty-five (50) female Sprague Dawley (SD) rats were divided into 5 groups. Group A was controlled while groups B, C, D, and E were treated groups. Group A received 0.5 ml of sterile water. Groups B, C, and D were given 0.2 mg/kg, 2 mg/kg, and 4 mg/kg respectively at diestrus. Group E was given 6mg/kg/day of CC dissolved in sterile water. The administration was done orally. A vaginal smear was evaluated for various estrus phases. ECR was calculated, and ova shed at the estrus phase was evaluated at autopsy. All data were presented as mean ± standard error of the mean (SEM). Statistical significance was taken at P &lt; 0.05&#x0D; Findings: There was a significant decrease (p&lt;0.05) in the number of proestrus, estrus, metestrus, and diestrus in treated groups compared with control. ECR value in all treated groups also decreased significantly (p&lt;0.05) when compared with control. This study further shows that all the animal groups (A-E) released ova but there was a significant decrease (p&lt;0.05) in the number of ova released in groups B, C, D, and E against the control group.&#x0D; Conclusion: CC decreases EC. It reduced ECR, hence prolonging luteogenesis. It decreases the number of eggs released. With a large dose and prolonged use, eggs were still released even when the ova released and counted were fewer compared with when this drug was not in use. &#x0D; Recommendation: Infertile couples should copulate more at the luteal phase of the menstrual cycle when on CC to increase their chances of pregnancy. CC decreases the number of eggs released but makes the few eggs released stay longer for fertilization.

The Role of Esophagography in the Diagnosis of Orthotopic Esophageal Squamous Cell Carcinoma in a Mouse Mode

Animal models play an irreplaceable role in pre-clinical studies of various tumors, mainly including xenogeneic/homograft subcutaneous tumor models. However, the subcutaneous transplanted tumor model is separated from the microenvironment of the primary tissue and fail to accurately simulate the growth of tumor cells in vivo. Therefore, orthotopic tumor models play an increasingly important role. For esophageal squamous cell carcinoma, chemotherapy-induced methods have successfully established mouse models of esophageal squamous cell carcinoma, but past experiments have found that it is still a challenge to confirm the success of the mouse model of esophageal cancer. The pathological diagnosis of the esophageal cancer tissue obtained from the killed mouse model remains the gold standard, but it also represents a lost opportunity to evaluate the efficacy of subsequent treatment. Notedly, esophagography is the primary examination for clinical patients with esophageal cancer. Therefore, the aim of this study is to evaluate whether esophagography can be used to establish a successful in situ mouse model of esophageal cancer. Referring to previous literature, we used the chemical drug 4-Nitroquinoline N-oxide (4-NQO) to induce orthotopic tumor model: A certain amount of 4-NQO was dissolved in propylene glycol to make a 2% stock solution, and then dissolved in 200ml of sterile water for the daily drinking water of mice. The water was kept in the dark and replaced once a week. After 16 weeks of induction, the drinking water was replaced by the same volume of sterile water and continued to be fed for 12 weeks. The mice were subsequently subjected to esophagography: First, the mice were fixed on a plastic plate, and the gavage needle was slowly inserted into the stomach, then slowly withdrawn out and gradually injected with meglumine diphosphate, with a total amount of 0.1-0.2ml. Then the radiolucent film was taken immediately, and three positions were selected, including the plain film, the standing film and the lateral film. After that, the mice were sacrificed and the esophagus of the mice was isolated and embedded in sections for pathological and immunohistochemical diagnosis. Finally, the results were compared with the imaging results. Based on the comparison of imaging and pathological results, we found that most of the esophageal segments with filling defects on esophagography were pathologically diagnosed as esophageal squamous cell carcinoma, and another part were diagnosed as esophageal papilloma. Compared with CT and MRI, esophagography has the advantages of simple operation and less consumable materials. We found that esophagography is a new non-invasive auxiliary diagnostic method for the successful establishment of an orthotopic mouse model of esophageal squamous cell carcinoma.

Implementation Status and Challenges Associated With Implementation of the Targeted Medication Safety Best Practices in a Tertiary Hospital.

Background The Institute for Safe Medication Practices (ISMP) is a well-known non-profit organization dedicated to preventing medication errors. Every two years they publish best practices that can reduce the occurrence of medication errors. This study aims to evaluate the implementation status of these best practices and to understand barriers associated with non-implementation at a tertiary hospital in Saudi Arabia. Methodology This was a two-phase qualitative study. First, a survey consisting of the ISMP best practices was sent to employees (mainly heads of departments) to fill out the implementation rate for each best practice. Then an interview or a focus group was conducted to further validate their answers and understand why some best practices were not implemented. Results Our study found that the highest implemented best practices were having strategies to improve safety with high-alert medications (best practice #19, 85.7%), having antidotes and reversal agents readily available (best practice #9, 75%), independent verification of sterile preparation (best practice #11, 75%), and limiting the number of removable medications from the automated dispensing unit by override (best practice #16, 75%). The least implemented best practices were ensuring that oral liquid medications are dispensed in a syringe (best practice #4, 12.5%), maximizing use of barcode verification (best practice #18, 12.5%), purchasing oral liquid dosing devices that display metric scale (best practice #5, 25%), eliminating glacial acetic acid from all areas of the hospital (best practice #6, 28.6%), and eliminating all 1,000 mL of sterile water from all areas outside of the pharmacy (best practice #10, 28.6%). Challenges associated with implementation were related to knowledge, motivation, and opportunity in the environment, with the latter being the highest barrier associated with non-implementation. Conclusions Healthcare providers need to have knowledge about the best practices and the rationale behind them, the motivation to perform them, and the necessary resources to implement the best practices in their hospital.

322. FLUORESCENCE LYMPHOGRAPHY USING INDOCYANINE GREEN IN THE PREVENTION OF CHYLE LEAKAGE AFTER MINIMALLY INVASIVE ESOPHAGECTOMY

Abstract Background Chyle leakage is one of the most challenging complications in esophagectomy, with an incidence of 14–21%. The clinical implications for patients are prolonged thoracic drainage and hospital admission, additional reinterventions, unpleasant dietary restrictions, and decreased long-term survival. The anatomy and vulnerability of the thoracic duct render it prone to injury, and mainly side branches and collaterals are often hard to visualize. Fluorescence lymphography using indocyanine green (ICG) can accurately visualize the thoracic duct, which may possibly reduce the incidence of chyle leakage and intra-operative detected chyle leakage can be treated directly. Methods A prospective observational cohort study is performed. Patients aged over 18 years with resectable (cT1-4a, N0–3, M0) esophageal carcinoma undergoing a minimally invasive esophagectomy are included. The primary endpoint of the study is postoperative chyle leakage. Secondary endpoints are overall morbidity and mortality, nature of dietary restrictions, reinterventions, hospital costs, 2 year survival rate and quality of life. Twenty-five milligram of ICG is dissolved in 10 mL of sterile water, obtaining a solution of 2.5 mg ICG per mL. In the McKeown esophagectomy, before the thoracic phase, the surgeon administers a bilateral bolus of 2 mL ultrasound-guided into inguinal lymph nodes. In an Ivor Lewis esophagectomy, 2 mL of reconstituted ICG will be injected into the small bowel mesenteric root just under the peritoneum of the mesentery, at the level where the jejunostomy is placed. Additionally, 50 mL of cream is enterally administered through the just established feeding jejunostomy. Subsequently, the operating area is inspected for chyle leakage with fluorescence lymphography after thoracic duct ligation and just before terminating the procedure. Results We aim to include 50 patients to assess the feasibility of fluorescent lymphography using ICG and enteral cream to prevent chyle leakage after minimally esophagectomy. Inclusion is ongoing, at this time, 31 of 50 patients are included. Results will be expected before the congress. Conclusion It is hypothesized that as a result of this technique the patients will benefit from lower incidence of postoperative chyle leakage, shorter thoracic drainage, reduced hospital admission, reduced dietary restriction, less re-interventions, and improved overall survival.

First Report of Root Rot Caused by Pleiocarpon algeriense on peony (Paeonia suffruticosa) in China.

Peony (Paeonia suffruticosa Andr.) is a perennial plant of Ranunculaceae. Its root bark (Danpi in Chinese) is a traditional Chinese medicine, which has the effects of clearing heat and cooling blood, promoting blood circulation to resolve blood stasis. Peony is mainly planted in the provinces of Anhui, Gansu, Henan and Shandong. Peony is also called Fengdan in the Fenghuang Mountain of Tongling, Anhui Province. In November 2021, a root rot-like disease was observed on the root of peony in several fields located in Tongling county, Anhui Province, China (118°0'51" N, 30°48'11" E). Approximately 20-40% of the peony plants were affected in the fields. The roots of the diseased plants were rotten and blackened, the bark of the roots was detached, and the leaves were withered, causing the whole plants to die. To isolate the pathogen, the symptomatic roots were sampled, and small pieces (5 × 5 mm) of diseased tissues were surface sterilized with 0.5% NaClO solution and 75% ethanol for 5 min, rinsed with sterile distilled water three times, and finally incubated on potato dextrose agar (PDA) at 28°C in the dark for 7 days. A total of 16 isolates were obtained from the infected tissues. Among isolates, six isolates were morphologically similar to B4. Colonies were passaged multiple times on fresh PDA medium, and pure isolate B4 exhibiting cinnamon-to-honey coloration on PDA with pale yellow aerial hyphae, was then selected. Microscopic observations revealed that microconidia were straight to curved, ellipsoid or subcylindrical, and ranged from 7.14 to 14.29 × 2.85 to 5.00 μm (n = 20). The morphological characteristics were similar to the description of Pleiocarpon algeriense by Aigoun-Mouhous et al. (2019). To further identify the taxonomic status of B4 strain, three genes of the internal transcribed spacer (ITS) region of rDNA, beta-tubulin (TUB2), and the RNA polymerase II second subunit (RPB2) were respectively amplified and sequenced using primers ITS1/ITS4 (White et al. 1990), T1/Bt-2b (O'Donnell and Cigelnik 1997), and 5F2/7cR (O'Donnell et al. 2007). Sequences for the isolate B4 were deposited in GenBenk (OP810684, ITS; OP882301, TUB2; OP863337, RPB2). BLAST analysis showed the ITS, TUB2, RPB2 sequences of B4 were 99.80% (505/506), 99.51% (609/612) and 100.00% (854/854) homology with those of P. algeriense Di3A-AP52 (MT613337, ITS; MT597145, TUB2; MT635004, RPB2). A phylogenetic tree was built using MEGA11 based on sequences of three genes showing that B4 strain was closely clustered with reference strain of P. algeriense, which has not been reported in peony in China. The pathogenicity test of the isolates was performed by inoculating 50 mL of conidial suspension (1 × 108 conidia/mL) on the roots of ten healthy peonies, ten peonies inoculated with 50 mL of sterile water were used as a control group. After one-month, typical symptoms of root rot appeared on the inoculated plants and the control plants were asymptomatic. The fungus (P. algeriense) was reisolated from the diseased roots and identified by sequencing of ITS gene, conforming to Koch's postulates. Pleiocarpon algeriense has been reported to cause stem and crown rot in avocado (Aiello et al. 2020). To the best of our knowledge, this is the first report of P. algeriense causing root rot in peony. Control methods of P. algeriense on peony fields will be studied in-depth in the future.