Effect of Low Temperature on the Aggressiveness of Rhizoctonia solani AG 2-2 Isolates on Sugar Beet (Beta vulgaris) Seedlings

Rhizoctonia solani anastomosis group (AG) 2-2 can cause seedling damping-off in sugar beets and substantial losses may occur in all regions where beets are grown. Sugar beets are planted early in the season when soil temperatures are low in order to maximize the length of the growing season and minimize the risk of damping-off. However, predictive models that indicate there is little to no risk of Rhizoctonia damping-off at temperatures <15°C may not be entirely reliable. We tested this possibility by inoculating sugar beet seedlings in a growth chamber at 11°C with 35 R. solani AG 2-2 isolates that were representative of the genetic diversity present in AG 2-2. Although disease progress and growth rate were greatly reduced at 11°C, considerable disease symptoms did develop in inoculated plants. Three weeks after inoculation, 16% of the plants were dead and 77% of the isolates tested had average disease severity scores that were significantly greater than those of the mock inoculated control. This confirms our concern about the possibility for low-temperature infection of sugar beets and indicates that waiting until the soil warms up to above 15°C to apply fungicide could leave the crop at risk. Aggressiveness does not appear to be related to subgroup or growth rate but rather depends on the response of the specific isolate to low temperature.

The Impact of Anastomosis Location in Patients with Lower Thoracic Esophageal Squamous Cell Carcinoma: A Retrospective Observational Study

BackgroundSeveral kinds of anastomoses with varying locations that can be performed after the surgical resection of lower thoracic esophageal squamous cell carcinoma. In this study, we evaluated the prognostic impact of anastomosis locations in these patients who underwent radical esophagectomy.MethodsLower thoracic esophageal squamous cell carcinoma patients which underwent radical esophagectomy and confirmed as microscopically complete resection were retrospectively enrolled. Anastomoses below the aortic arch or below the azygos arch were defined as low anastomosis. Other anastomoses were defined as high anastomosis. Overall survival of these two kinds of anastomoses were analyzed using the log-rank test and Cox regression model.ResultsOf the 781 patients enrolled, 196 and 585 were classified as the low anastomosis and high anastomosis groups, respectively. Overall, the survival time in low anastomosis group (median OS, 36.1 versus 65.4; P=0.01) was shorter than high anastomosis group but no statistical difference was observed in multivariate analysis (P=0.195). Again, no significant difference in survival between low anastomosis and high anastomosis group (median OS, 140.9 versus 124.8; P=0.345) were observed in pT1-T2 subgroup. In pT3-T4 subgroups, patients with low anastomosis group had significantly poorer survival that those with high anastomosis (median OS, 27.1 versus 42.9, P=0.003), even after controlling for other confounders (P=0.026). Notably, the impact of anastomosis location on long-term survival in pT3-4 patients was not significantly modified by nodal status. The internal validation of patients undergoing Sweet approach shown that pT3-T4 patients with high anastomosis had survival advantages (adjusted HR=0.711, 95%CI, 0.601 0.990, P=0.041)ConclusionsFor lower thoracic esophageal squamous cell carcinoma with declared T3-4 status, low anastomosis is associated with worse prognosis and should be avoided.

A Blumgart-Anastomosis Based Half-Invagination Pancreaticoenterostomy With Better Applicability to Laparoscopy and Lower Incidence of Pancreatic Leakage

Background The Blumgart anastomosis has been established as one of the safest anastomoses for pancreas remnant reconstruction, with low rates of postoperative pancreatic fistula (POPF) and postoperative complications. However, how to make laparoscopic pancreaticoenterostomy easier and safer is still a subject to be discussed. Methods Data of patients undergoing laparoscopic pancreaticoduodenectomy from April 2014 to December 2019 were retrospectively analyzed. Results 20 cases of half-invagination anastomosis (Group HI) and 26 cases of Cattell-warren anastomosis (Group CW) were included. Intraoperative blood loss, Operative time and Postoperative drainage tube placement time of Group HI was significantly less than those of Group CW. Patients of Clavien-Dindo grade III and above in Group HI was significantly less than Control Group. Group Incidence of POPF in Group HI was significantly lower than that in Group CW. No high-risk group emerged during Fistula risk score analytical phase, and the highest risk of moderate-risk group was pancreatic leakage. Respectively, incidence of pancreatic leakage in Group HI and Group CW was 7.7% and 46.67%, while incidence of Group HI was significantly lower than that in the Group CW. Conclusions The Blumgart-anastomosis based half-invagination pancreaticoenterostomy with better applicability to laparoscopy can effectively reduce the incidence of postoperative pancreatic leakage.

Genome Analyses of the Less Aggressive Rhizoctonia solani AG1-IB Isolates 1/2/21 and O8/2 Compared to the Reference AG1-IB Isolate 7/3/14

Rhizoctonia solani AG1-IB of the phylum Basidiomycota is known as phytopathogenic fungus affecting various economically important crops, such as bean, rice, soybean, figs, cabbage and lettuce. The isolates 1/2/21 and O8/2 of the anastomosis group AG1-IB originating from lettuce plants with bottom rot symptoms represent two less aggressive R. solani isolates, as confirmed in a pathogenicity test on lettuce. They were deeply sequenced on the Illumina MiSeq system applying the mate-pair and paired-end mode to establish their genome sequences. Assemblies of obtained sequences resulted in 2092 and 1492 scaffolds, respectively, for isolate 1/2/21 and O8/2, amounting to a size of approximately 43 Mb for each isolate. Gene prediction by applying AUGUSTUS (v. 3.2.1.) yielded 12,827 and 12,973 identified genes, respectively. Based on automatic functional annotation, genes potentially encoding cellulases and enzymes involved in secondary metabolite synthesis were identified in the AG1-IB genomes. The annotated genome sequences of the less aggressive AG1-IB isolates were compared with the isolate 7/3/14, which is highly aggressive on lettuce and other vegetable crops such as bean, cabbage and carrot. This analysis revealed the first insights into core genes of AG1-IB isolates and unique determinants of each genome that may explain the different aggressiveness levels of the strains.

Single Layer Extra-Mucosal Versus Double Layer Intestinal Anastomosis for Colostomy Closure: A Prospective Comparative Study

Background: The main objective was to compare the outcome of single layer interrupted extra-mucosal sutures with that of double layer suturing in the closure of colostomies. Subjects and Methods: Sixty-seven patients with closure colostomy were assigned in a prospective randomized fashion into either single layer extra-mucosal anastomosis (Group A) or double layer anastomosis (Group B). Primary outcome measures included mean time taken for anastomosis, immediate postoperative complications, and mean duration of hospital stay. Secondary outcome measures assessed the postoperative return of bowel function, and the overall mean cost. Chi-square test and student t-test did the statistical analysis.. Results: Thirty-two patients were allocated to group A and 35 patients to group B. The mean time taken for anastomosis was significantly shorter in group A (23.25 ± 1.20 min in group A vs. 36.71 ± 1.93 min in group B; P<0.001). A significant shorter duration of hospital stay was seen in group A (7.00 ± 1.778 days in group A vs. 9.74 ± 1.990 days in group B; P<0.001). The detection of bowel sound was substantially quicker in group A as compared to group B (4.56 ± 0.50 days in group A vs. 6.46±0.50 days in group B; P<0.001). There was no significant discrepancy between the two groups regarding anastomotic leak rates (P= 0.543). The mean cost of double layer intestinal anastomosis method was significantly higher than that of single layer anastomosis (P<0.001). Conclusions: The use of single layer extra-mucosal anastomosis of the intestine has the advantage of taking less time, less morbidity and cost-effective to perform with the same rate of anastomotic leak in the closure of colostomy.

Liver transplantation in pediatric patients under 15 kg; duct-to-duct vs. Roux-en-Y hepaticojejunostomy biliary anastomoses

Back ground. Liver transplantation is an effective treatment for acute or chronic liver failure and metabolic liver disease, which is associated with good quality of life in over 80 percent of recipients. We aimed to evaluate outcome of duct-to-duct vs. Roux-en-Y hepaticojejunostomy biliary anastomoses in pediatric liver transplant recipients below 15-kg.Methods. In this single-center retrospective study, all children less than 15 kg that have undergone liver transplantation at Nemazee Hospital Organ Transplant Center affiliated with Shiraz University of Medical Sciences from 2009 till 2019, were enrolled. Over a 10-yr period, 181 liver transplants were performed in patients with two techniques including duct-to-duct (Group 1) vs. Roux-en-Y hepaticojejunostomy biliary anastomoses (Group 2). All data was collected from patients’ medical records, operative notes, and post-transplant follow up notes. Data was analyzed by SPSS software V21.Results. Overall, 94 patients had duct to duct anastomosis (group 1) and 87 cases had Roux-en-Y hepaticojejunostomy (group 2). The mean age of the patients was 2.46 ± 1.5. The most common underlying diseases was biliary atresia (32%). The most prevalent complication after the surgery was infection in both groups. cardiopulmonary problems were significantly higher in group 2 (24.1% vs 4.3%) (p < 0.001). The rate of infection was significantly higher in group 2, as well.Conclusion. Our study showed a relatively high rate of post-operative infection which was the most among patients who had undergone Roux-en-Y hepaticojejunostomy. Except from biliary complications which were mostly observed in DD group, other complications were more common among Roux-en-Y group.

First report of collar and root rot of faba bean caused by Rhizoctonia solani AG-2-2 IIIB in Bangladesh

Faba bean (Vicia faba L.) is an underutilized promising grain legume commercially grown in central and northern part of Bangladesh (Yasmin et al. 2020). In January 2021, faba bean plants exhibiting symptoms of collar and root rot and yellowing of leaves were observed in thirty plots of an experimental field at the Bangladesh Agricultural University (24.75° N, 90.50° E), Mymensingh, Bangladesh. Infected plants had dark brown to black lesions on the roots, extending above the collar region. An average disease incidence and severity was 7.16% and 6.91%, respectively. Eight diseased plants were collected from the field by uprooting one plant from each of eight randomly selected experimental plots and surface disinfected with sodium hypochlorite (0.2%) for 3 min followed by 1 min in ethanol (70%), and then rinsed three times with distilled water and dried on sterile paper towels. Collar and root pieces (5×5 mm) of symptomatic tissues were placed on Potato Dextrose Agar (PDA). Plates were incubated at 25°C for three days and isolates were purified from single-tip culture. The isolates produced brown colored mycelia often with brown sclerotia. Under microscope, fungal colonies exhibited right–angled branching with constriction at the base of hyphal branches and a septum near the originating point of hyphal branch consistent with the description of Rhizoctonia solani Kuhn (Sneh et al. 1991). The isolates grew at 35°C on PDA (5 mm/24). Molecular identification of the isolates BTRFB1 and BTRFB7 was determined by sequencing the rDNA internal transcribed spacer (ITS) region using primers ITS1 and ITS4 (White et al. 1990). A BLAST search showed that the sequences (GenBank Accession nos. MZ158299.1 and MZ158298.1) had 99.28% similarity with R. solani isolates Y1063 and SX-RSD1 (GenBank Accession nos. JX913811.1 and KC413984.1, respectively). Phylogenetic analysis revealed that the present isolates grouped with R. solani anastomosis group AG-2-2 IIIB. To confirm pathogenicity, both isolates were grown individually on sterile wheat kernels at 28°C for 6 days (D’aes et al. 2011). Faba bean seedlings were grown in plastic pots containing sterile potting mix (field soil/composted manure/sand 2:2:1 [v/v]). Two-week-old plants were inoculated by placing five infested wheat seeds adjacent to the roots. Control pots were inoculated with sterile wheat kernels using the same procedure. Plants were placed in a growth room with a 16 h/8 h light/dark photoperiod at 25 ± 2°C after inoculation. Fifteen days after inoculation, typical collar and root rot symptoms were developed on inoculated plants, similar to symptoms observed in the field. Control plants remained non-symptomatic. Finally, six isolates of R. solani were isolated from the symptomatic plants and identified by morphological and molecular analysis. Rhizoctonia solani is the causal agent of seed and root rot, hypocotyl canker, and seedling damping-off diseases of faba bean in many other countries (Rashid and Bernier 1993; Assunção et al. 2011). To our knowledge, this is the first confirmed report of Rhizoctonia solani causing collar and root rot of faba bean in Bangladesh. This finding will be helpful for the development of management strategies to control this disease and to expand the production of faba bean in Bangladesh.

Effect of Ozone on Apelin and APJ Expressions in Rat Peritonitis-Constituted With Colon Anastomosis

Background: Analyze the effect of ozone therapy on Apelin and APJ expressions in peritonitis constituted colon anastomosis.Methods: Eighteen male Wistar albino rats weighing 250-300 g were used in this study. The rats were randomly assigned into three groups. In the colonic tissue samples Hematoxylin-Eosin staining (HE) and the Apelin and APJ immunostaining was applied. Also, Apelin and APJ protein levels between groups were determined with the Western-Blot method.Results: In the ozone therapy group, Apelin and APJ immunoreactivity was decreased compared to the anastomosis group. The protein levels of Apelin and APJ according to Western-Blot analysis are consistent with immunostaining. Conclusion: As a result, increased levels of Apelin and APJ in cecal punctuation and colonic anastomosis process can be deduced and said to contribute to worsening of tissue while it may be involved in return to normal of with treatment.

First Report of Soybean Seedling Disease caused by Rhizoctonia solani AG-7 in Taiwan

From August to November 2020, reduced emergence and damping-off of soybean seedlings were observed in two fields (Benzhou and Wandan) in Taiwan. Disease incidence was approximately 40% in Benzhou by field scouting. The roots of damping-off seedlings were brown. Affected seedlings could be easily pulled out from the soil and the lesions on the roots/stem were generally dry and sunken. These symptoms suggested the possibility of Rhizoctonia infection. Soil surrounding symptomatic seedlings were collected to bait the potential pathogen and symptomatic plants were used for pathogen isolation. The diseased tissues were washed with tap water and surface-disinfected with 1% bleach before placing on the Dexon selection medium at 26°C for 2 days (Ko and Hora 1971). Hyphae were transferred to potato dextrose agar (PDA), and a brown colony with brown and irregular-shaped sclerotia grew from 90 out of 99 isolates. The hyphae exhibited typical characteristics of Rhizoctonia solani, including a constriction and a septum near the end of branching hyphae (Ajayi-Oyetunde and Bradely, 2018). Two isolates from Benzhou and two isolates from Wandan were tested for their pathogenicity, and eight surface-disinfected seeds were distributed evenly on the water agar plates covered by 2-day-old mycelia at 25°C in dark for 7 days. All isolates caused cotyledon rot and reduced germination. To verify their pathogenicity in pots, double-sterilized sorghum seeds were inoculated with two strains and incubated at 25°C for 2 weeks to be used as fungal inoculum (Ajayi-Oyetunde and Bradely, 2017). A layer of 15 ml of fungal inoculum was placed 5 cm beneath the soil surface in pots. Four soybean seeds were planted approximately 3 cm above the inoculum in each pot. After two weeks, reddish lesions on the hypocotyls or taproots of all seedlings in the inoculated pots were observed, while seedlings in the control pots inoculated with sterile sorghum seeds remained healthy. The pathogen was re-isolated from lesions and had identical morphology to the original isolates. To characterize the fungal identity, the internal transcribed spacer (ITS) was sequenced using the primers ITS1/ITS4 (Sharon et al., 2006). Using BLASTN in the NCBI database, the sequence (GenBank no. MW410857 and MW410858) showed 100% (639/639 bp) similarity to KF907734 and 99.83% (635/636 bp) similarity to AF354099, both belong to R. solani anastomosis group 7 (AG-7) (Hua et al. 2014; Gonzalez et al. 2001). Phylogenetic analysis comparing sequences with different AGs (Ajayi-Oyetunde and Bradely, 2017) grouped our isolates within the AG-7 clade with a 100% bootstrap confidence. In the anastomosis test, an incompatible zonation and unequal mycelial growth rates were observed when AG-7 isolates were paired with an AG-1 IA isolate. On the other hand, the compatible tuft reaction was observed when two AG-7 isolates were paired, and the compatible merge reaction was observed in the self-pairing tests (Macnish et al. 1997). Accordingly, the molecular and morphological characterizations confirmed the causal pathogen as R. solani AG-7. R. solani AG-7 was first reported on radishes in Japan (Homma et al., 1983), first found on carnation in Taiwan (Lo et al., 1990), and in field soils of various crops but not soybean (Chuang, 1997). It was suggested that Rhizoctonia diseases of soybean may be present in Taiwan, but molecular confirmation was lacking (Anonymus, 1979). As R. solani AG-7 causes diseases of soybean in the US and Japan (Baird et al., 1996), the importance of AG-7 as an endemic pathogen of soybean in Taiwan should be recognized and its prevalence determined as a first step to managing this disease.

Application value of hand-sewn anastomosis in totally laparoscopic total gastrectomy for gastric cancer

Background Digestive tract reconstruction in totally laparoscopic total gastrectomy can be divided into two types: instrument anastomosis and hand-sewn anastomosis. This study explored the feasibility and safety of hand-sewn sutures in esophagojejunostomy of totally laparoscopic total gastrectomy, compared with instrument anastomosis using an overlap linear cutter. Methods This retrospective cohort study was conducted from January 2017 to January 2020 at one institution. The clinical data of 50 patients who underwent totally laparoscopic total gastrectomy, with an average follow-up time of 12 months, were collected. The clinicopathologic data, short-term survival prognosis, and results of patients in the hand-sewn anastomosis (n=20) and the overlap anastomosis (n=30) groups were analyzed. Results There were no significant differences between the groups in sex, age, body mass index, American Society of Anesthesiologists score, tumor location, preoperative complications, abdominal operation history, tumor size, pTNM stage, blood loss, first postoperative liquid diet, exhaust time, or postoperative length of hospital stay. The hand-sewn anastomosis group had a significantly prolonged operation time (204±26.72min versus 190±20.90min, p=0.04) and anastomosis time (58±22.0min versus 46±15.97min, p=0.029), and a decreased operation cost (CNY 77,100±1700 versus CNY 71,900±1300, p<0.0001). Postoperative complications (dynamic ileus, abdominal infection, and pancreatic leakage) occurred in three patients (15%) in the hand-sewn anastomosis group and in four patients (13.3%) in the overlap anastomosis group (anastomotic leakage, anastomotic bleeding, dynamic ileus, and duodenal stump leakage). Conclusion The hand-sewn anastomosis method of esophagojejunostomy under totally laparoscopic total gastrectomy is safe and feasible and is an important supplement to linear and circular stapler anastomosis. It may be more convenient regarding obesity, a relatively high position of the anastomosis, edema of the esophageal wall, and short jejunal mesentery.