Suspension In Distilled Water Research Articles

Comparison of different molecular protocols for the detection of Uncinaria stenocephala infection in dogs

The present study aims to assess the performance of different molecular targets using various matrices of samples for the detection of Uncinaria stenocephala (US) in hookworm infected dogs. To this end, the DNA extraction was performed on the following matrices of samples: (i) larvae of US obtained from experimentally infected dogs with US with different larvae counts per microliter (µl); (ii) pure US eggs suspension in distilled water with different egg counts per µl; (iii) spiked dog fecal samples with different US eggs per gram (EPG) of feces; (iv) feces from dogs naturally infected with hookworm eggs; (v) fecal suspension with hookworm eggs recovered from the FLOTAC apparatus. All the samples were tested with four different PCR protocols targeting specific regions for the detection of both hookworms US and AC as follows: Protocol A (ITS1, 5.8 S, ITS2) and Protocol B (18 S) for the detection of both species, Protocol C (ITS1) for the detection of AC and Protocol D (ITS1) for the detection of US. The best results were obtained with DNA extracted from US larvae matrix obtained from experimentally infected dogs, showing a detection limit of 3.5 larvae/ml for the protocols A, B and D. A moderate correlation was found between the FLOTAC technique and PCR protocols B and D with respect to fecal samples from dogs naturally infected with hookworms. Indeed, PCR protocols B (18 S) and D (ITS1) gave the best results for feces and fecal suspension from naturally infected dogs. However, all the PCR protocols used showed lower sensitivity than FLOTAC technique. Perhaps, isolating US eggs in advance could help to obtain better quality and quantity of DNA, avoiding some notable factors such as inhibitors present in faecal samples. However, a further study is needed to evaluate and standardise a protocol for the recovery of parasitic elements, that could be applied prior to DNA extraction. Therefore, this could lead to a better amplification of US eggs DNA. In conclusion, our results showed that the type of sample (sample-matrix) used for the DNA extraction samples is crucial, as this affects the diagnostic sensitivity of the technique.

Assessment and mapping of soil salinity and groundwater quality in Nijhum Island, Hatiya Upazila, Noakhali

Crop yield is heavily influenced by soil salinity. The objective of the research is to assess the depth wise soil salinity distribution pattern and groundwater quality by measuring physico-chemical parameters in the agricultural land of Nijhum Island, Noakhali, Bangladesh. This study also explicates the spatial variation of soil salinity in the study area. Soil samples were collected from three different depths to estimate the Electrical Conductivity (EC) using a paste of 1:5 soil and distilled water suspension (1:5 weight-to-volume method). Spatial analyst tool of ArcGIS was used and observed that the land of Nijhum Island was affected with severe salinity in almost 85% of sampling plots and 15% of sampling plots were extremely saline. The soil salinity process has been noticed due to seawater intrusion from the Bay of Bengal through the Meghna estuary which threatens crop production and soil fertility, threatening the population's livelihood. This study provides a baseline understanding of soil salinity in Nijhum Island to help decision-makers and smallholder farmers improve their livelihoods. Thus, groundwater depth did not affect soil salinity significantly. The result contradicts most studies that found a strong influence on soil salinity. Because the study area is coastal, most of the salt comes from seawater inundation and capillary rise, which reduces groundwater availability.

A Rapid and Robust DNA Extraction Method for PCR-Based Diagnosis of V. cholerae

Cholera, caused by _V. cholerae_, needs rapid diagnosis because of its threat for rapid spread. Recent molecular diagnosis by PCR assay is more advantageous than the conventional methods. The bottleneck in PCR diagnosis lies in the delay in template DNA extraction of _V. cholerae_, which is the basic requirement. This study describes a simple, less expensive, and rapid template DNA extraction method to lessen the total time for PCR diagnosis of _V. cholerae._ To obtain template DNA, our method involves boiling of _V. cholerae_ suspension in distilled water (takes 18-24 hours) instead of boiling the suspension in broth medium, which is a lengthy process (takes 72 hours). The validation of our template DNA was conducted using 40 _V. cholerae_ O1 strains that were confirmed previously, and its usefulness was checked on 20 clinical strains of _V. cholerae_ O1 isolated from acute diarrhea patients; results were compared with those of conventional template DNA. The analysis of our template DNA in simplex and quadruplex PCR assays revealed the presence of _ctxA_, _tcpA _(El Tor), _rfb _(O1), _ToxR_, _ctxB_, _zot_, _ace,rst _, and _OmpW _genes in _V. cholerae _O1. The comparison of the results of PCR assays using the template DNA from both sources showed equal results and matched correctly. The sensitivity of our template DNA was 1.5x103 CFU per assay. Our template DNA extraction method is simple, less expensive, and rapid, and it can be employed for early diagnosis of _V. cholerae_ during outbreaks, in research laboratories, and in hospital setups where infrastructures are available.

Evaluation of the possibilities of using albumin microspheres for radioaerosol study of the deposition of inhaled substances and mucociliary clearance

The need to study and evaluate the leading protective mechanisms of the respiratory system remains one of the key, but unresolved problems of modern medicine, concerning both radiation hygiene and toxicology, and pulmonology. The model of the respiratory tract presented by the ICRP only partially addresses them, significantly limiting the feasibility and possibilities of its use. The solution of this problem undoubtedly contributes to the improvement of methods of prevention and treatment from the effects of incorporative radionuclides entering by inhalation, as well as methods for diagnosing and treating respiratory diseases aimed at the earliest stages of their pathogenesis, given that the processes of deposition of inhaled substances and lung clearance directly depend both therapeutic and pathogenic effects. This is especially relevant when there is a danger of contamination with radioactive waste, when there is an urgent need to deal with highly pathogenic influence and its consequences. The most informative method for studying the processes of deposition inhalant and mucociliary clearance is radioaerosol. However, due to the circumstances, there are no radiopharmaceuticals that could be used for such a study since the 2000s, which prompted us to carry out this work. At the same time, albumin microspheres labeled with 99mTc are considered potentially suitable, but not studied radiopharmaceuticals in this regard. The aim of the work was to study the aerodynamic properties of albumin microspheres and evaluate the possibilities of their use for radioaerosol study of the processes of deposition of inhaled substances and mucociliary clearance. The aerodynamic properties of albumin micro[1]spheres were studied using optical and scanning electron microscopy by studying their dry dispersion, initial and obtained from its suspension in distilled water, generated by an ultrasonic inhaler according to the developed technology. The results indicated that the dispersible particles were characterized by sphericity, clear boundary differentiation and a relatively homogeneous distribution of particles with a relatively narrow size range (spread 1-2.5 µm) and little conglomeration, as well as a smooth surface. Conclusion. The investigated albumin microspheres are suitable for radioaerosol study of the processes of deposition of inhaled substances and mucociliary clearance.

Development of laboratory experiments to determine critical moisture condition of CLT constructions

There is an increased interest in using Cross-laminated timber (CLT) in construction, but many buildings are erected without weather protection, which poses a risk of moisture impact if wood is exposed to precipitation during construction. The construction industry argues that there are no documented critical moisture levels for CLT and no specific test method. In the study, a laboratory test set-up was developed to study mould growth under realistic and controlled climatic conditions after exposure to distilled water and spore suspension. In the experiments, small test specimens of CLT structures were exposed to distilled water for 1 day or 1 week. During the development of the method it was found that exposed for one day and then given the opportunity for open drying did not give rise to mould growth. On the other hand, growth occurred on surfaces that could not dry immediately, for example at connection points. For specimens exposed for one week, mould growth arose regardless of whether the surfaces could dry immediately or not. The conclusions apply primarily to the climates studied. The methodology needs to be further developed, with other scenarios being studied, and calibrated against samples exposed to outdoor air, dust, dirt and rainwater.

First report of Phakopsora euvitis causing leaf rust disease on grapevine (Vitis labrusca L.) in India.

Grapevine (Vitis labrusca L.), a member of the family Vitaceae and native of North America, is grown as a table grape. During the survey for the grapevine diseases in May 2022, we noticed numerous yellow pustules of rust on lower side of leaves of 'Bangalore Bule' in Nandi village (13°22'59.7"N77°42'33.4"E), Chikkaballapur district of Karnataka state of India. The crop was at maturity stage and the rust disease severity was determined using the scale given by Angelotti et al. (2008), which was up to 10%. The disease symptoms were numerous small raised yellow pustules on the abaxial surface corresponding to adaxial surface chlorotic spots. In severe conditions, spots cover the entire leaf and defoliation occurs. Similar disease symptoms were reported by Ono (2000); Weinert et al. (2003); and Primiano et al. (2017). The pathogenicity test was performed on cuttings of 'Bangalore Bule' grapevine in a glasshouse at 25±1°C. The urediniospores were collected from diseased leaves using a brush, 3×104 ml-1 suspension in distilled water was used for inoculation on the abaxial surface of leaves. Control plants were sprayed with distilled water. The leaves developed symptoms in 15-17 days after inoculation, and the pathogen was confirmed by symptomatology and microscopic observation of urediniospores. Urediniospores were short-pedicellate, sessile, obovoid to obovoid-ellipsoid, and uniformly echinulate with 42.98-32.54 x 31.37-25.15 µm in size. The aecial stage of the Phakopsora has been reported on an alternate host, Meliosma simplicifolia (Hosagoudar 1988). As the internal transcribed spacer (ITS) region offers some utility in the molecular detection of the Phakopsora genus (Rush et al. 2019), the pathogen was confirmed by studying the different regions in the ITS such as ITS1, 5.8S rRNA, and ITS2. Total DNA was extracted from urediniospore mass using the Macherey-Nagel kit (Duren, Germany) by following the manufacturer's protocol. The quantity of isolated DNA was checked using an Qubit 3.0 fluorometer (Invitrogen) before being subjected to polymerase chain reaction (PCR) amplification in a thermocycler (Eppendorf-vapo.protect) using ITS1 and ITS4 primers (IDT, Singapore) targeting ITS1, 5.8S rRNA, and ITS2 regions, and the obtained amplicon (~700 bp) was purified using Macherey- Nagel Nucleospin gel and PCR clean-up kit (Duren, Germany) as per the manufacturer's protocol and sequenced by Sanger's dideoxy chain-termination method [ABI 3730 (48 capillaries) electrophoresis]. The sequence was edited in BioEdit (https://bioedit.software.informer.com/7.2/) and aligned in MUSCLE, and the phylogenetic tree was constructed in MEGA 11 using the neighbor-joining method by following the maximum likelihood criterion (Kumar et al. 2018). The sequence data was deposited at NCBI (accession number OP221661). The basic local alignment search tool (BLAST) search sequence of the isolate Nandi-KA in GenBank revealed 97.91% homology with sequence of Phakopsora sp. (accession number KC815548.1) and 96.87% with Phakopsora euvitis (accession number AB354790.1). Based on disease symptoms, fungal morphology, the pathogenicity test, and ITS sequence, the fungus was identified as Phakopsora euvitis, the pathogen causing grapevine leaf rust disease. Though there are similar disease symptoms observed on grapevine in India (EPPO 2016); the pathogen was not confirmed. To our knowledge, this is the first report of Phakopsora euvitis causing leaf rust disease in grapevine (V. labrusca) in India.

Hepatoprotective Potential of Phytoconstituents and Vitamins against Antitubercular Drugs Induced Hepatotoxicity in Albino Rats

Background: This study aims to develop a new hepatoprotective drug from common plants and vitamins that are potent, nontoxic, and cost-effective. The literature search found that Curcuma longa, Zingiber officinale, Terminalia, chebula, and Vitamin A (Vitamin C + Vitamin E) provide hepatoprotective action against drugs-induced hepatotoxicity. A major side effect of antitubercular drugs (ATD) is liver toxicity, which reduces their effectiveness. Aim: In this study, Vitamins and Phytoconstituents (Combined extract) were evaluated for their potential hepatoprotective effects against hepatotoxicity induced by antitubercular drugs in Wistar albino rats of either sex. Materials and Methods: In the study, ethanolic extract of rhizomes of Curcuma longa and Zingiber officinale, fruits of Terminalia chebula, and vitamins C and E were used. As a standard drug, silymarin was also used. For 30 days, albino rats received 7.5 mg/kg isoniazid, 10 mg/kg rifampicin, and 35 mg/kg pyrazinamide orally as a suspension in distilled water. Result: A combined extract plus vitamins (500mg/kg) treatment significantly reduced the hepatic toxicity caused by antitubercular drugs (P<0.05-P<0.001). A combination of extracts + vitamins (500mg/kg) eliminates hepatotoxicity, and the results are close to those of Silymarin, a standard drug. Conclusion: As a result of this study, extracts+vitamins provide protection against liver injury attributed to their hepatoprotective activity, which supports their traditional use.

Development and Experimental Evaluation of Some Silver Nanoparticles with Antimicrobial Potential

By adjusting the synthesis process, silver nanoparticles (AgNp) of various shapes, sizes, and structures can be obtained, all of which have a substantial impact on the biological effect, notably, the regulation of antibacterial activity in the present circumstances of growing bacterial resistance. Due to their relatively small size, nanoparticles may be disseminated evenly throughout the body of the experimental animal, even at low doses, and exert more potent antibacterial activities. Our research was centered on the synthesis, production, and biological evaluation of antibacterial silver nanoparticles. Using the Turkevich method, we were able to effectively synthesize and characterize nanoscale silver particles, with an average crystallite size of 9.49 nm. We examined their acute toxicity and pharmacokinetic characteristics in rats after administering a single dosage. In addition, we evaluated the biological effect of topical AgNp suspension on the progression of burn-type lesions in the experimental animals. The pharmacokinetic profile demonstrated that the plasma concentration of silver nanoparticles, as well as their clearance rate, and dispersion throughout the body, are significantly enhanced in large rodent species. The restorative effect of synthesized silver nanoparticles in the form of a suspension in distilled water was corroborated by the values of the hematological parameters. These results demonstrated an intense stimulation of the cellular and molecular processes of the local immune defense, which has resulted in significantly faster regeneration in the AgNp-treated group.

Морфологическая характеристика внутренних органов мышей при длительном потреблении микропластика

Introduction.Microplastics (MP) pollute the environment and can pose a danger to living organisms including humans. Experimental studies on mice and rats demonstrated that microplastics can enter the internal environment, causing structural damage to and dysfunction of various organs. However, data on this issue are scarce and contradictory. The aim of this paper was to characterize morphological changes in the internal organs in mice after prolonged MP consumption in different doses. Materials and methods. We formed four groups (n=5 each): three experimental and one control group. Mice of the experimental groups were given a 5-μm polystyrene particle suspension in distilled water at a concentration of 0.1, 1, and 10 mg/l for 4 weeks. The daily consumption doses averaged 0.023, 0.23, and 2.3 mg of microparticles per kg of animal body weight, respectively. The control group received distilled water throughout the experiment. We conducted a morphological examination of the colon, spleen, liver, kidneys, lungs, thymus, mesenteric lymph nodes, brain, heart, testicles, and thigh muscles. In the colon, morphometric methods were used to evaluate inflammatory infiltration, the number of endocrine and goblet cells, and the content of neutral and highly sulfated mucins in the latter. Results. On morphological examination under the influence of 5-μm polystyrene particle consumption in different doses, we revealed pathological changes only in the spleen and colon. However, a decrease in the goblet cell volume fraction in the colon mucosa was found in the group with low dose of microplastics (0.023 mg/kg/day). The group that consumed 2.3 mg/kg/day of microplastics showed the activation of both spleen compartments, an increased endocrine cells count, and a decreased proportion of highly sulfated mucins in goblet cells. Conclusion. Polystyrene microparticles with a 5-μm diameter consumed for 4 weeks at doses of 0.023–2.3 mg/kg/day cause adaptive morphological and functional changes in the colon and spleen. Keywords: microplastics, polystyrene, spleen, colon, internal organs

Experimental research on droplets releasing characteristics of the bubble bursting behavior at a free surface with an aerosol

Bubble burst phenomena exist widely in industrial and environmental processes and exert a subtle influence. Bubbles may be produced by gas injection or boiling and will burst after floating at the free surface of the pool for a while. The produced film droplets and jet droplets will entrain the liquid substance into the gas, especially in reactor accidents. When aerosols retain in the pool, radioactive aerosol-laden droplets may exert a long-lasting source term that needs to be addressed to evaluate this source more precisely. The bubble bursting and film droplets producing behavior at the free surface under different conditions are investigated by visualization. Image processing software is applied to measure the size, number, and speed of film droplets. By comparing the bubble cap film thickness and the film roll-up speed to the number of droplets under different liquid temperatures both in distilled water and aerosol suspension, we show that the increase of the cap film thickness (decrease of the bubble lifetime) attributes to the decline of the number and average speed of film droplets and the increase of average size. The normalized droplet speed obeys the multimodal distribution. The normalized droplet size distribution obeys the gamma distribution and the Rayleigh distribution. The goodness of fit for gamma distribution of distilled water can reach 88%, and aerosol suspension is 70%. The increase of the liquid temperature will slightly increase the number and average size of film droplets but decrease the average speed for distilled water and aerosol suspension. Under low temperature, adding TiO2 aerosol may increase the number of film droplets; under high temperature, the aerosol may reduce the number of droplets.

Highly antimicrobial and strong cellulose-based biocomposite film prepared with bacterial cellulose powders, Uncaria gambir, and ultrasonication treatment

This work characterized bacterial cellulose (BC)/Uncaria gambir (G) biocomposite film prepared with ultrasonication treatment. Films were prepared from BC powder suspensions in distilled water without and with various loadings (0.05 g, 0.1 g, 0.2 g, 2 g) of G powder then treated using an ultrasonic probe at 1000 W for one hour. The results revealed that the ultrasonication treatment of the suspension greatly increased tensile strength (TS), elongation at break (EB), and toughness (TN) of a BC film by 3097%, 644%, and 32,600%, respectively, compared to non-sonicated BC film. After adding 0.05 g G into the sonicated BC powder suspension, TS, EB, and TN of the biocomposite film were improved to 105.6 MPa, 14.3%, and 8.7 MJ/m3, respectively. The addition of the G increased in antimicrobial activity of the film. This study indicates that biocomposite film is potentially useful for nanopaper production with good antimicrobial and high tensile properties.

Konya İli Böğürtlen, Ahududu ve Kuşburnu Yetiştiriciliğinde Potansiyel Bir Tehdit: Ateş Yanıklığı Hastalığı

In the present study, totally 49 samples, which showed the symptoms of leaf and shoot blight and cankers with brown discoloration of necrotic tissues on mature branches, were collected from 22 districts and areas of Konya Province between 2017 and 2019. Presence rate of E. amylovora in collected samples, showing symptoms of the disease, from the province was determined to be 40% for blackberry and raspberry and 33% rosehip for rosehip in three years. Bacteria consistently isolated from the diseased tissues were identified on the basis of biochemical, physiological, and molecular tests, comparing with a reference strain of E. amylovora, isolated from blackberry (Kbb 371). Twenty seven representative bacterial strains were gram-negative, rod-shaped, mucoid, fermentative, positive for levan formation and acetoin production, no growth at 36°C, positive for gelatin hydrolysis, and negative for esculin hydrolysis, indole, urease, catalase, oxidase, arginine dehydrolase, reduction of nitrate, acid production from lactose, and inositol. All strains induced a hypersensitive response in tobacco (Nicotiana tobacum cv. White Burley) 24 h after inoculation with a 108 CFU ml-1 bacterial suspension in sterile distilled water. The strains were identified as E. amylovora using the species-specific primers set A/B (1), which amplified a 1-kb DNA fragment in PCR, and the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) method. In order to fulfill the Koch postulates, pathogenicity test was confirmed by injecting bacterial suspensions of 108 CFU ml-1 in sterile distilled water into the shoot tips of 3-year-old blackberry R. fruticosus cv. Chester, raspberry R. idaeus cv. Heritage and rosehip R. canina. All tests were repeated three times. The bacterium was re-isolated from inoculated plants and identified as E. amylovora. Phytosanitary measures are needed to prevent any further spread of the bacterium as potential inoculum sources to new blackberry, raspberry and rosehip growing areas.

Influence of Inorganic Metal (Ag, Cu) Nanoparticles on Biological Activity and Biochemical Properties of Brassica napus Rhizosphere Soil

Two separate forms of application of silver and copper nanoparticles (AgNP and CuNP in a nanocolloidal suspension) to winter oilseed rape seeds were used: (1) seed soaking (S) for 1 h at 20 °C in a NP suspension and (2) additional seed soaking and spraying plants 21-day-old seedlings (SP) with NP. The AgNP and CuNP colloidal suspensions in sterile distilled water were applied in three different NP concentrations (50, 100, 150 mg L−1). However, the changes in the biology and biochemistry of the Brassica napus rhizospheric soil after the application of CuNPs and AgNPs are not considerable, although mostly statistically significant, and the application of CuNPs is more beneficial for this activity than the application of AgNPs. The number of CFUs (colony–forming units) of the tested groups of culturable microorganisms (fungi and copiotrophic, oligotrophic, and siderophore-producing bacteria) indicates the following trend: the abundance of all the tested groups was slightly positively correlated with CuNPs and clearly negatively correlated with AgNPs in each version of application. The soil pH value and tested biochemical soil parameters (IAA: indole-3-acetic acid, PhC: phenolic compounds, FeCC: Fe–chelating compounds) were negatively correlated with AgNPs applied to the seeds (S) at all the tested concentrations and to the seeds and plants (SP) at the concentration of 50 mg L−1. In turn, these parameters were strongly positively correlated with CuNPs applied to the S and SP groups at the concentration of 50 mg L−1 as well as Ag applied to SP at 100 mg L−1. Decrease in dehydrogenase activity (DHA) was lower after the application of CuNPs and AgNPs in S than in the SP way, and the DHA activity was equal to the activity in the control sample after the CuNP application in 100 and 150 mg L−1 concentrations.

Stemphylium lycopersici Causing Leaf Spot of Watermelon (Citrullus lanatus) in China.

Watermelon (Citrullus lanatus) is an important cucurbit crop in China. During September 2020, an unknown leaf spot disease was observed onwatermelon in two greenhouses (640m2 per greenhouse) of Sangzi town, Jizhou district, in Tianjin,China(117°10'E, 39°55'N), where approximately 10% of plants were infected. Disease symptoms began as small, circular, brown spots on leaves. As these spots increased in size, they developed confluent, irregular lesions surrounded by dark brown edges. Severely affected plants had many wilted leaves followed by defoliation. Ten symptomatic leaves were collected for pathogen isolation. Diseased tissues (3×3 mm) were cut from the margins of lesions and surface disinfected with 1% NaClO for 1 min, rinsed three times with sterile distilled water and then placed on potato dextrose agar (PDA) at 25±2°C with a 12-h photoperiod for 7 to 10 days. Seven morphologically similar isolates were obtained from the ten infected leaves and purified by single-spore culturing for further study. The initial growth of the isolates on PDA appeared grayish white in obverse and bright yellow pigmentation in reverse. Colony color gradually deepened to grayish brown in obverse and brownish red in reverse. Conidia (n=50) were solitary, light brown, oblong to long elliptic, pointed or obtuselyrounded atthetop, constricted at the transverse septum, with verrucous processes on the surface, 36.3 to 64.2×16.6 to 25.1 μm, and the L/W ratio of conidia was 1.5-2.5. All characteristics were consistent with the description of Stemphylium lycopersici (Ellis 1971; Woudenberg et al. 2017). Total genomic DNA was extracted from a representative isolate (XG2-2) using a Fungal DNA Kit (GBCBIO, Guangzhou, China). The internal transcribed spacer (ITS) and translation elongation factor 1-α (EF1-α) genes (Sun et al. 2015) were amplified and sequenced with the primer pairs ITS1/ITS4 (5'-TCCGTAGGTGAACCTGCGG-3'/5'-TCCTCCGCTTATTGATATGC-3') and EF-1α-F/EF-1α-R(5'-TCACTTGATCTACAAGTGCGGTGG-3'/5'-CGATCTTGTAGACATCCTGGAGG-3'), respectively. The two sequences of strain XG2-2 (GenBank Accession No. MW362344 and MW664941) showed 100% and 99% identity toS. lycopersici strain 01 and strain KuNBY1 (GenBank Accession No. KR911814 and AB828256) respectively. The phylogenetic analysis using MEGA7 based on the sequences of ITS and EF1-α regions showed that the isolate XG2-2 was clustered withS. lycopersiciisolates (strain 01 and strain KuNBY1). For the pathogenicity test, a spore suspension (1×106 spores/ml) in sterile distilled water from a 7-day-old culture of the fungus grown on PDA and counted with a hemacytometer was sprayed on leaves and stems of five healthy watermelon plants, grown for 2 months in the greenhouse at 25 to 30 °C, with 85% relative humidity. Conditions remained the same for inoculation experiments. Negative controls were healthy plants inoculated with sterile distilled water. The experiment was repeated twice. Six days after inoculation, typical leaf spot symptoms were observed on inoculated leaves, whereas control leaves remained symptomless. To satisfy Koch's postulates, the causal fungus was re-isolated from the lesions of inoculated plants, with morphological and cultural characteristics identical with the original isolate. Stemphylium lycopersici is a common fungus with a relatively extensive host range (Kee et al. 2018). In recent years, new host plants infected by S. lycopersici have been reported in Asia including Physali (Yange et al. 2020), common bean (Li et al. 2019), and potato (Kee et al. 2018). To our knowledge, this is a new host record for S. lycopersici causing leaf spot on watermelon in China. Sangzi watermelon is a special local product in the Jizhou district of Tianjin. At present the cultivated area in 1000 ha including 667 ha in controlled conditions and 333 ha of field-grown plants with a total annual output of 45000 Mg. In this survey, we found the disease caused by S. lycopersici on watermelon only in these two greenhouses, but cannot rule out the possibility of large-scale spread in the future. Therefore, integrated management strategies for this fungus need to be developed to reduce economic losses in commercial cultivation.

Efficacy of standardized novel Boswellia serrata extract in the dextran sodium sulfate-induced colitis model - potential use in gut health management

Background: Objective of this study was to evaluate anti-inflammatory properties of a novel standardized Boswellia serrata extract–bsRx (developed using natural excipients and designed to have specific ratio of its major actives, viz. AKBA and BBA) in dextran sodium sulfate (DSS)-induced IBD model in BALB/c mice.Methods: Animals (BALB/c mice) in control (CL) group were administered vehicle; DSS-induced colitis group (DSS group), 2.5 % DSS; and Boswellia serrata group (BS group) received DSS, for inducing colitis, together with a novel standardized extract of Boswellia serrata (41 mg/kg, 4.1 mg/ml solution in distilled water) for 10 days. Reference group (SS group) received DSS with sulfasalazine (30 mg/kg, 3.0 mg/ml suspension in distilled water) for 10 days. Clinical assessment for disease activity index (DAI), histopathological examination and hematological assessments were performed.Results: Treatment with Boswellia serrata showed significant reduction in the DAI score on day 10 compared to the DSS group (2.49±0.93 versus 3.63±0.55, p≤0.05). Body weight (18.54±2.21 gm versus 17.05±3.53 gm) and colon length (6.8±0.9 cm versus 7.6±0.6 cm, p≤0.05) also improved in the BS group compared to DSS group, respectively. Histological scoring of colitis was lower in the BS group (10.1±1.37). There was no difference in leukotriene levels between groups (p>0.05).Conclusions: Treatment with novel Boswellia serrata extract improved colon length, DAI and histological scoring index in DSS-induced colitis in IBD mice models. Our results indicate the promising potential of novel Boswellia extract in IBD and gut health management.

First Report of Fusarium Root and Stem Rot Caused by Fusarium oxysporum f. sp. radicis-cucumerinum on Greenhouse Cucumbers in Saudi Arabia.

Cucumber (Cucumis sativus L.) is an important vegetable crop in Saudi Arabia. During May 2018, 45 - 60% of 5-month-old cucumber plants showed symptoms of a previously unknown wilt in commercial greenhouses around Al Kharj area of Riyadh region. Symptoms consisted of crown and root rot, wilting and stem disintegration, along with yellowish brown to brown external discoloration extended throughout the affected tissues. As the disease progressed, a pinkish-orange mycelial growth was often observed at the basis of affected stems while vessels were discolored. Subsequently, the affected plants were collapsed and died. Crown, stem, and root fragments (4 × 4 mm) were cut from symptomatic tissues, surface sterilized in 2.5% NaOCl, cultured on potato dextrose agar (PDA) with 25 mg/liter of streptomycin sulfate, and incubated at 26°C in darkness for 6 days. Single-spored cultures produced white mycelium with pink, white, or purple pigmentation in the center. The mycelium produced sporodochia. Macroconidia were mainly slightly curved with three to five septa. Microconidia were single-celled oval and produced on short lateral phialides. Chlamydospores were single or in short chains. Morphologically, the isolated fungus was characterized as Fusarium oxysporum (Leslie and Summerell 2006). To further confirm the fungus identification, DNA was extracted from a single-spored culture. Three different fungal nuclear regions of internal transcribed spacer (ITS), elongation factor 1-α, (TEF1-α) and the second largest subunit of DNA-directed RNA polymerase II (rpb2) with the following primers: ITS4 and ITS5 (White et al. 2017), EF-1 and EF-2 (O'Donnell et al. 2008), and fRPB2-5F and fRPB2-7cR (Liu et al. 1999), respectively. The ITS, TEF1-α, and rpb2 sequences of the isolate FCKSU17 were submitted to GenBank (MT232918, MW471131, and MW449833 respectively). Phylogenetic analysis based on the alignment of the ITS, TEF1-α, and rpb2 sequences using MEGA7 placed this strain in the F. oxysporum clade. To confirm the forma specialis radicis-cucumerinum, amplification with the specific primers ForcF1/ForcR2 was conducted (Lievens et al. 2007). The amplified fragment (∼ 250-bp) was sent for sequencing, and the sequence was submitted to GenBank (MW471132). BLASTn analysis of the sequences showed 100% identity with F. oxysporum radicis-cucumerinum (KP746408). To fulfill Koch's postulates, pathogenicity test was conducted on 7-day-old plants of cucumber cultivar Beit Alpha grown into pots filled with soil mix (2:1 sandy loam-peat moss, vol/vol). The plants were inoculated through drenching with 100 ml of conidial suspension in sterile distilled water (106 spores/ml) per pot. Control plants were treated with sterile distilled water. Each treatment included 10 replicates (pots), with two plants per pot. The pathogenicity test was repeated once. Cucumber plants inoculated with the fungus showed early wilting symptoms within the first 2 weeks post inoculation. At the 6th week post inoculation, 90 to 100% of the inoculated plants developed typical symptoms. No symptoms were observed on the control plants. The pathogen was successfully re-isolated from the inoculated wilted plants and identified morphologically. To our knowledge, this is the first report of F. oxysporum f.sp. radicis-cucumerinum on cucumber in Saudi Arabia. It is recommended that preventive management should be considered as this disease may cause significant economic losses on cucumbers in Saudi Arabia.

First Report of Postharvest Fruit Rot Disease of Hardy Kiwifruit Caused by Diaporthe eres in China.

Hardy kiwifruit (Actinidia arguta), as an economically important fruit crop growing in Northeast China with thin, hairless and smooth skin, is susceptible to postharvest decay. In September 2018, infected cultivar Kwilv fruits were obtained from a commercial farm in Liaoning province, northeastern China. The occurring incidence of the rot disease varied from 20% to 90% according to the fruit number in each box during a 7-day-long storage at room temperature, and the initial symptom included a small, soft, chlorosis to light brown lesion and later watery brown lesions. Pure cultures of the same characteristics were obtained from the isolated strains in four rotten fruits on PDA medium. The isolates grew into transparent radial mycelium on PDA in the first two days followed by abundant white, fluffy aerial mycelium. After 14 days, colonies formed white to light brown aerial mycelial mats with gray concentric rings, and they produced gray and embedded pycnidia. Alpha conidia of 4.4 to 8.8 µm × 1.4 to 3.3 µm (n = 50) were abundant in culture, hyaline, aseptate, ellipsoidal to fusiform, while Beta conidia at 20.5 to 28.6 µm × 1.0 to 1.4 µm (n = 50) were hyaline, long, slender, curved to hamate. These morphological characteristics were similar to Diaporthe species (anamorph: Phomopsis spp.) (Udayanga et al. 2014). For identification, DNA was extracted from three single isolates respectively , and the internal transcribed spacer (ITS) region, β-tubulin (BT), and histone (HIS) H3 gene were amplified by using primers ITS1/ITS4 (White et al. 1990), T1/T22 (O'Donnell et al. 1997) and HIS1F/HISR (Gao et al. 2017), respectively. The three isolates produced identical sequences across all three gene regions, which were submitted to NCBI (Genbank accession numbers MT561361, MT561360 and MT855966). Nucleotide BLAST analysis revealed that the ITS sequence shared 99% homology with those of ex-type Diaporthe eres in NCBI GenBank (MG281047.1 and KJ210529.1), so did the BT sequence that had 98% identity to D. eres (MG281256.1 and KJ420799.1) and the HIS 99% identity to D. eres (MG28431.1 and MG281395.1) (Hosseini et al. 2020, Udayanga et al. 2014). Pathogenicity was tested by wound inoculation on the cv. Kwilv fruits. Five mature and healthy fruits were surface-sterilized with 1% NaClO solution, rinsed in sterile distilled water and dried. Every fruit was wounded by penetrate the peel 1-2 mm with a sterile needle, and inoculated with mycelium plugs (5 mm in diameter) of the isolate on PDA, with five inoculated with sterile PDA plugs as controls. Treated fruits were kept in sterilized transparent plastic cans separately under high humidity (RH 90 to 100%) at 28°C. After five days, the same rot symptoms were observed on all fruits inoculated with mycelium while the control remained symptomless. The fungi was re-isolated from the lesions of inoculated fruits and identified as D. eres by sequencing, thus fulfilling Koch's postulates. The pathogenicity experiment was re-performed using D. eres conidial suspension (107 conidia/ml) in sterile distilled water in October 2019 and the same results were obtained. D. eres was recently reported to cause European pear rot in Italy (Bertetti et al. 2018). To our knowledge, this is the first report of D. eres causing a postharvest rot in hardy kiwifruit in China, leading to severe disease and thus huge economic losses in Northeast China. Accordingly, effective measures should be taken to prevent its spreading to other production regions in China.

A simple method for the extension of shelf life of cultures of Phytophthora species causing black pod disease of cacao (Theobroma cacao L.)

Black pod disease of cacao caused by Phytophthora palmivora and Phytophthora megakarya in Ghana take heavy toll of cacao production in the field. Intensive research has been carried out worldwide on these pathogens. However, viability of the cultures during prolonged storage has remained a major challenge in the research. This paper reports findings of assessment of six storage media viz sterilised distilled water (SDW), sterilised and unsterilized soil suspension (SSS and USS), vegetable 8 juice broth (V8JB), Oat Meal Agar slant under mineral oil (at 4°C) and empty tube. Viability of the cultures was assessed on V8JA and in tetrazolium chloride test. Ability of zoospores of the cultures to infect cacao leaf discs was used to assess growth vigour and pathogenicity. Phytophthora cultures stored in SDW (26 ± 2°C; alternating day light and night) were preserved for 60 days (5 years). Both P. palmivora and P. megakarya performed better on SDW and SSS than on USS due to removal of staling substances in the soil medium by the sterilisation. Vigour of growth and pathogenicity of the stored cultures required re-inoculation of host tissue (cacao pod) in order to maintain potency to continually infect host.

The inhibiting effect of microwave radiation on Paenibacillus larvae spores suspended in water

The aim of this paper was to investigate the effects of microwave radiation on the viability of Paenibacillus larvae spores and to study the relationship between the microwave power consumption, the exposure time and the number of spores in the examined suspensions. Sterile distilled water suspensions were made using larval detritus, to contain tens, hundreds and thousands of spores. The suspensions of all the dilutions were gradually exposed to a microwave radiation power of 170, 510 and 850 W. In all the cases, the exposure time was 1, 2, 3, 4, 5, 10, 15, 20, 25 and 30 minutes. After cooling, 0.1 ml of each exposed suspension was inoculated onto three modified MYP (mannitol egg yolk polymyxin) agar plates and incubated aerobically at 37 ± 1 °C for 120 hours. The statistical evaluation of the spore counts decreasing with time was performed with the use of the nonparametric Friedman’s variance test using the Unistat Statistical Package v6.5. The results showed that the rate of devitalisation of the spores is dependent on the microwave oven power consumption, but independent of the number of spores. Using a power consumption of 170, 510 and 850 W, the devitalisation of the spores occurred after 15, 3 and 2 min of exposure, respectively.

Stability of Reconstituted Amoxicillin and Potassium Clavulanate Oral Suspensions Marketed in the Democratic Republic of the Congo

Amoxicillin and Clavulanate potassium oral suspensions are one of antibiotics combination mainly available as dry powders for reconstitution. Made of a β-lactam molecule with a β-lactamase inhibitor, this combination existence can be justified by the potency of Clavulanate potassium which protects Amoxicillin against bacteria producing β-lactamases. Once reconstituted, suspension in single dose bag need to be taken by patient on daily basis following the medical prescription and the one in bottle to be taken according to the medical prescription and the remaining part (in the same container) kept refrigerated in order to get the optimal benefit from the drug. This study investigated the stability of Amoxicillin and Clavulanate potassium suspension both in bags (sachets) for single dose per daily-usage and in bottles for multiple dose usage. The last were reconstituted using distilled water and stored in refrigerator and the previous were instantly reconstituted before measurement. The stability evaluation was conducted on three brands (Six samples totally, two per brand including bottle and sachet for each brand) and for duration of seven days using a validated HPLC method. The evaluation was based on the measurement of pH values, color observation of reconstituted suspensions stored under refrigerator during 7 days and the active compounds concentrations determination. After analysis, no change was observed in pH values and a small change of color for multiple dose presentations stored in refrigerator. And the concentrations of suspensions evaluated in day 1 and day 7 showed over 90% up to seventh day under same storage conditions for both of the Amoxicillin and Clavulanate potassium pharmaceutical presentations. As the study aimed to assess the stability of the suspension in single-dose from bag presentation and in multiple-dose from suspension in bottle, we observed that the reconstituted suspension in distilled water of Amoxicillin + Clavulanate potassium stored in refrigerator is stable for seven days of use, independently from their pharmaceutical presentations either for oral suspensions (powders) in bottles or those in bags for single use after reconstitution.