Antifungal Activity of Shirazi Thyme (Zataria multiflora Boiss.) Essential Oil against Hypomyces perniciosus, a causal agent of wet bubble disease of Agaricus bisporus

Wet bubble disease (WBD) caused by Hypomyces perniciosus is a major constraint of button mushroom (Agaricus bisporus) cultivated worldwide. A few synthetic chemical fungicides are used to control WBD. In our study, the potential of essential oil (EO) from Zataria multiflora in inhibition of H. perniciosus was evaluated as an alternative to chemical fungicides. An isolate of H. perniciosus was isolated from wet bubble diseased A. bisporus and pathogenicity of the mycoparasite was determined under artificially inoculated conditions. The mycoparasitic fungus was identified using sequences of the internal transcribed spacer (ITS) region of ribosomal DNA. The EO was extracted from the aerial parts of Z. multiflora by microwave extraction method and evaluated in vitro for its antifungal activity against H. perniciosus. The EO of Z. multiflora (ZEO) at the tested concentrations (50% and 100%) inhibited the growth of H. perniciosus in the agar diffusion test. The minimum inhibitory concentration (MIC) of ZEO was 0.04% as assessed by the poisoned food technique. The chemical composition of ZEO was determined by gas chromatography-mass spectrometry analysis. A total of 23 compounds were identified. Among them, the most abundant compounds were Linalool (20.3%) and Bornyl acetate (15.5%). Linalool at the tested concentrations of 0.25% and 0.125% completely inhibited the mycelial growth of H. perniciosus in an in vitro assay. These results suggest that ZEO can be exploited for control of WBD

Assessment of aflatoxin B1 content and aflatoxigenic molds in imported food commodities in Muscat, Oman

Aflatoxins, mainly produced by Aspergillus flavus and A. parasiticus are considered as serious food safety and human health issues due to their hepatotoxic effects. In the present study, the occurrence of aflatoxin B1 (AFB1), the most potent human liver carcinogen, and prevalence of toxigenic isolates of Aspergillus spp. were assessed in 140 food commodities in Muscat markets, Oman, and the 95 quarantined imported food commodities. These samples consisted of rice, corn, peanut, red chilli powder, soybean, dried dates and tree nuts. AFB1 was analyzed using competitive ELISA/LC-MS and the aflatoxigenic fungi were detected using plating technique followed by molecular identification. No AFB1 was detected in 89 (63.6%) samples collected from local markets, while 44 (31.4%) samples contained 1-5 ppb and the remaining 7 (5%) samples (red chili powder) contained 6-10 ppb. None of the samples exceeded the maximum permissible limit of 10 ppb set for foods by Oman legislation. Of the 95 quarantined samples, only 17 (17.9%) samples were positive and contained AFB1 at concentrations ranging from 1-3.4 ppb. Four isolates of Aspergillus pp. were isolated from the collected samples and were identified as Aspergillus flavus (A14, A16 and A23) and A. chevalieri (A46) on the basis of internal transcribed spacer (ITS) sequences of ribosomal DNA. Among them, A. flavus strain A14 alone produced AFB1 (7.6 ppb), while A16, A23, and A46 were non-toxigenic. This is the first detailed report on the occurrence of AFB1 in food commodities imported into Oman

Development and morphological changes in leaves and branches of acid lime (Citrus aurantifolia) affected by witches’ broom

Witches’ broom (WB), associated with the presence of ‘Candidatus Phytoplasma aurantifolia’, is one of the most serious diseases of acid lime. This study determined incidence, distribution, and development of the disease, and morphological changes in leaves and branches of affected host plants. Survey in different parts of Oman showed that WB occurs in most regions in the country, where 108 out of 158 (68%) surveyed farms were found to have diseased trees. A survey of 6,926 acid lime trees showed that severity of WB was positively related (r = 0.948; P<0.01) to tree age. The mean percentage of symptomatic branches was 1% in 3-year-old trees compared to 63% in 12-year-old trees. To further characterize morphological changes in WB-affected limes, apical stems (40 cm long) were collected from three infected trees during the autumn of 2009 and spring of 2010. Increases in the numbers of leaves (1,208%), numbers of branches (309%) and total length of branches (712%) were recorded for symptomatic branches relative to non-symptomatic branches. In the spring of 2009 these respective increases were 159%, 243% and 121%.Overall area of leaves in the symptomatic branches was 81% less than for non-symptomatic branches in the autumn of 2009 and 34% less in the spring of 2010. This study is the first to characterize morphological changes in leaves and branches of acid lime affected by WB

Witch’s Broom Disease of Lime (Candidatus Phytoplasma aurantifolia):Identifying High-Risk Areas by Climatic Mapping

Biological invasions of vectorborne diseases can be devastating. Bioclimatic modeling provides an opportunity to assess and predict areas at risk from complex multitrophic interactions of pathogens, highlighting areas in need of increased monitoring effort. Here, we model the distribution of an economically critical vectorborne plant pathogen ‘Candidatus Phytoplasma aurantifolia’, the etiological agent of Witches’ Broom Disease of Lime. This disease is a significant limiting factor on acid lime production (Citrus aurantifolia, Swingle) in the Middle East and threatens its production globally. We found that temperature, humidity, and the vector populations significantly determine disease distribution. Following this, we used bioclimatic modeling to predict potential novel sites of infections. The model outputs identified potential novel sites of infection in the citrus producing regions of Brazil and China. We also used our model to explore sites in Oman where the pathogen may not be infectious, and suggest nurseries be established there. Recent major turbulence in the citrus agricultural economy has highlighted the importance of this work and the need for appropriate and targeted monitoring programs to safeguard lime production

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

The effect of salt-tolerant antagonistic bacteria from tomato rhizosphere on plant growth promotion and damping-off disease suppression under salt-stress conditions

A total of 24 morphologically distinct salt-tolerant bacteria were isolated from the tomato rhizosphere soil and tested in vitro against Pythium aphanidermatum, causing damping-off disease in tomato. Among them, five bacterial isolates viz., BTR1.0, BTR1.1, BTR4.4, BTR7.0 and BTR8.6 were found to inhibit the mycelial growth of P. aphanidermatum. The maximum antagonistic effect was shown by BTR1.0 followed by BTR7.0 and BTR1.1. Based on the MALDI Biotyper analysis, these bacterial isolates were identified as Serratia marcescens (BTR1.0 and BTR1.1), Pseudomonas aeruginosa (BTR7.0 and BTR8.6) and Alcaligenes faecalis (BTR4.4). Analysis of plant growth stimulating activity of bacterial strains revealed that A. faecalis BTR4.4 recorded the highest vigour index followed by S. marcescens BTR1.0. The biocontrol activity of bacterial antagonists against tomato damping-off was tested under salt-water irrigation. Among the bacterial strains tested, S. marcescens BTR1.1 was found to be highly effective under non-saline water irrigation, which suppressed the disease by 68% compared to infected control. However, at 50 mM NaCl concentration, S. marcescens strain BTR1.0, P. aeruginosa strain BTR7.0 and A. faecalis BTR4.4 provided a significant level of control of the disease. At 100 mM NaCl concentration, no significant effect of the bacterial strains on the disease incidence was observed

Etiology, development and reaction of muskmelon to vine decline under arid conditions of Oman

Muskmelon vine decline is considered the most important factor limiting muskmelon production in Oman. This study was conducted to characterize incidence, development, causal agents and response of muskmelon cultivars to this disease. A survey showed that incidence of the disease ranged from 0 to 15% (mean 5%) in spring 2011, 1 to 80% (mean 18%) in autumn 2011 and 0 to15% (mean 10%) in spring 2012. Isolations from 168 affected plants yielded Pythium aphanidermatum (56% of diseased plants sampled), Fusarium spp. (46%), Monosporascus cannonballus (27%), Rhizoctonia solani (22%) and Macrophomina phaseolina (1%). In pathogenicity tests, R. solani, M. cannonballus and P. aphanidermatum were found to be pathogenic to muskmelon. In another experiment over three seasons, M. cannonballus, P. aphanidermatum and R. solani were consistently isolated from muskmelon plants on a weekly basis from 14 days after sowing until the end of the season. However, symptom development only began with the onset of fruiting, which suggests that fruiting stress may be a factor in vine decline disease development. Field assessment of 11 muskmelon cultivars showed that ‘Shahd F1’ was one of the cultivars least susceptible to vine decline and was relatively high yielding. This is the first record of M. cannonballus as a causal agent of muskmelon vine decline in Oman

Potential of indigenous antagonistic rhizobacteria in the biological control of Monosporascus root rot and vine decline disease of muskmelon

Monosporascus root rot and vine decline (MRVD) of muskmelon, caused by Monosporascus cannonballus, is an economically important disease worldwide. The objectives of this study were to isolate native rhizobacterial strains and to evaluate their ability to promote plant growth and to provide protection against M. cannonballus. Thirty eight native bacterial isolates from the rhizosphere soil of muskmelon and cucumber were screened for their antagonism against M. cannonballus in a dual culture assay. Among them, five isolates viz., Bacillus amyloliquefaciens (B4), Pseudomonas mendocina (B7), Bacillus endophyticus (B10), Pseudomonas resinovorans B11 and P. aeruginosa AT3, identified based on the 16S rRNA gene sequence analysis, showed a significant level of antagonism and the inhibition zone ranged from 5.6 mm to 25.9 mm. Scanning electron microscopic observation of the hyphae of M. cannonballus at the inhibition zone revealed morphological abnormalities including shrinkage, loss of turgidity, pit formation and deformation. These bacterial isolates showed compatibility with one another and with Trichoderma viride. Greenhouse experiments revealed that P. resinovorans B11 was the most effective among the bacterial antagonists in controlling MRVD in melon. When applied as seed treatment and soil application, P. resinovorans B11 reduced the incidence of MRVD by 93.1%, relative to the infected control