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MethodsX 2022•To bypass the problem of viable but non-culturable bacteria that cannot be isolated by culturable methods would be to isolate DNA from bacterial cells concentrated...
•To bypass the problem of viable but non-culturable bacteria that cannot be isolated by culturable methods would be to isolate DNA from bacterial cells concentrated from water samples used as a template for the polymerase chain reactions (PCR). DNA extraction protocol (Omar et al. 2010) was used as a foundation for extracting DNA from water. The method combinations i.e., guanidium thiocyanate, celite and home-made spin column were chosen because it has been shown to be reliable, rapid, simple, and inexpensive for routine analysis in developing country settings.•The following optimizations were included: (a) Polycarbonate (Poly) was statistically compared with Polyether sulphone (PES), Nitrocellulose acetate (NA) and Nitrocellulose (NC) membranes; (b) Various housing containers for the membranes were tested: plastic/glass petri-dish, Falcon tubes, Ogreiner cryovials; (c) various solutions was tested to add to the membrane to remove cells from membranes; (d) celite was chosen to bind the DNA because it had a higher DNA binding capacity compared to silicon dioxide; (e) incubation times and rotation speed were tested when adding reagents.•The optimized in-house DNA extraction method was validated with environmental water samples, high (dam water) and low (borehole) bacterial load to determine upper and lower detection limits of the method.
PubMed: 35313544
DOI: 10.1016/j.mex.2022.101653 -
Journal of Dental Research, Dental... 2023This study evaluated the cytotoxicity of four bioceramic root canal sealers (RCSs) in vivo. The embryonic zebrafish characteristics, such as mortality, survival,...
BACKGROUND
This study evaluated the cytotoxicity of four bioceramic root canal sealers (RCSs) in vivo. The embryonic zebrafish characteristics, such as mortality, survival, hatching, and general morphology, served as the parameters for assessing cytotoxicity.
METHODS
The RCSs, namely GuttaFlow Bioseal, MTA Fillapex, CeraSeal Bioceramic, and iRoot SP, were mixed according to the manufacturer's guidelines. The extract solution was prepared by immersing the set RCS into 1X dilution of E3 solution. Then, the extract solution was delivered into a Petri dish where zebrafish embryos were allowed to develop. Cytotoxicity was evaluated 24, 48, 72, and 96 hours after fertilization.
RESULTS
The Kruskal-Wallis test showed that except for GuttaFlow Bioseal, the mortality, survival, and hatching of zebrafish embryos for the remaining three bioceramic RCSs were significantly different from the negative controls (<0.05). Significant differences were also evident in the mortality, survival, and hatching of zebrafish embryos between GuttaFlow Bioseal and three other RCSs (<0.05).
CONCLUSION
GuttaFlow Bioseal was less cytotoxic than other bioceramics RCSs; MTA Fillapex, CeraSeal Bioceramic root canal sealer, and iRoot SP root canal sealer exhibited comparable cytotoxicity.
PubMed: 38584992
DOI: 10.34172/joddd.2023.39163 -
Poultry Science Feb 2022Dust present in poultry houses can disseminate bacteria in air and deposit them on surfaces. This study evaluated bacteria in settled dust during growout of broilers...
Dust present in poultry houses can disseminate bacteria in air and deposit them on surfaces. This study evaluated bacteria in settled dust during growout of broilers from 2 flocks (Flocks A and B). Dust samples for bacteria analyses were obtained during 6 wk of growout (Flocks A and B) and 1 wk after flock termination (Flock B) by environmental swabbing and collecting dust in petri dishes from multiple locations inside the poultry house. For weekly swabbing, dust deposited during each wk of the sampling period (noncumulatively, n = 12/wk) and cumulatively (n = 12/wk) throughout the sampling period was collected. Swabbed dust samples were analyzed for counts (log CFU/28 cm) of aerobic bacteria, E. coli, coliforms, and Salmonella recovery. For petri dish dust collection, dust was collected in weekly and bi-weekly time spans during the sampling period and then analyzed for Salmonella recovery. Data were analyzed by one-way ANOVA and Fisher's Exact Test and means were separated using LSD. Only aerobic plate counts changed over time in dust during growout (Flocks A and B; P < 0.0001). In noncumulatively settled dust, aerobic bacteria (Flocks A and B; P < 0.0001), E. coli (Flock A; P = 0.0432), and coliforms (Flock B; P = 0.0303) varied during growout with peak counts on wk 5 or wk 6, wk 4, and wk 4, respectively, after bird placement. Salmonella recovery did not vary in cumulatively (3/72, 10/84) and noncumulatively (0/12, 10/84) settled dust during growout in both flocks. In dust sampled by bi-weekly collection in petri dishes, Salmonella recovery was highest (5/6) between wk 2 to wk 4 for Flock B (P = 0.0118). Overall, this study displayed that settled dust bacteria levels can fluctuate during broiler growout, and dust can contain Salmonella.
Topics: Animals; Chickens; Dust; Escherichia coli; Poultry Diseases; Salmonella Infections, Animal
PubMed: 34936956
DOI: 10.1016/j.psj.2021.101602 -
Plants (Basel, Switzerland) Nov 2022In this study, the chemical composition of the essential oils (EOs) obtained from different aerial parts (flowers, leaves, and stems) of Guss., a wild species endemic...
In this study, the chemical composition of the essential oils (EOs) obtained from different aerial parts (flowers, leaves, and stems) of Guss., a wild species endemic of Sicily, was investigated. Furthermore, the EOs' biocidal effects towards two pests of stored products, and , were evaluated. This activity was evaluated in Petri dish bioassays to establish the survival rate of adults treated with the EOs comparing them with solvent and a commonly used insecticide (pyrethrum). The data obtained from the toxicity bioassay evidenced that stems' EOs and leaves' EOs have a contact/fumigation effect towards the two insect species tested, while the EOs from the flowers did not exhibit a different mortality than the solvent. The EOs from the stem and leaves of , tested at 10 mg/petri dish, determined a LT50 of 53.38 and 42.97 h, respectively, on adults, and of 45.23 and 42.97 h, respectively, on adults. The promising bioactivity of leaves' EOs and stems' EOs toward and is encouraging in the perspective to test these oils and their main constituents for further experiments in the laboratory and field.
PubMed: 36432776
DOI: 10.3390/plants11223047 -
Plant Disease Mar 2021The panicle branch, which is the key node for transport of photosynthesis products from source to sink, is vulnerable to many diseases caused by fungal pathogens, such...
The panicle branch, which is the key node for transport of photosynthesis products from source to sink, is vulnerable to many diseases caused by fungal pathogens, such as Magnaporthe oryzae, Cochliobolus miyabeanus. Among these diseases, rice blast is the most important one which causes devastating losses in many regions. In 2019 and 2020, panicle branch rot of rice with a symptom which could be mistaken with rice blast was observed in a paddy field, where is not traditional epidemical region of rice blast, in Fuyang, Zhejiang province. In 2020, similar symptom was also observed in Hubei and Anhui Province. In a paddy field in Fuyang, the symptom appeared on more than 30% investigated panicles. Diseased panicle exhibited brown to black lesions on primary or secondary branches as well as pedicels, however the grain and the neck of spike could not be infected which is the most obvious difference with rice blast. Obviously, the disease can't destroy the entire function of branch and blank grain was rarely observed, so its damage is not comparable with neck blast. Normally, it caused incomplete grain filing commonly leading to 5% - 25% grain weight loss. During the booting stage of rice, local solar irradiation time and temperature were fewer and lower than common years which may be responsible for losses caused by this disease. After surface sterilized, lesion parts cut from infected branches from 25 panicle samples were cultured on 2% water agar at 28℃ for 24-28 h, and fungi were isolated and purified by mycelial tip transferring. Among 31 isolates, 26 showed similar cultural characters. The wool-like mycelia were luxuriant and grew rapidly on PDA spreading the whole 9 cm petri dish in less than a week at 28 ℃. The mycelia were white to ashen at beginning and gradually turned black from center of the plate after 5 days culture at 28 ℃. Hyphae were smooth, branched, septate, hyaline or pale brown. Conidia were single-celled, black, spherical to subspherical, and 10.2 to 14.6 × 12.2 to 15.7 μm (n=50) in dimension and born on tip of hyaline and ampulliform conidiophores. The fungus showed similar morphological characteristics with Nigrospora oryzae (1). ITS sequences of 6 representative strains of the fungus were amplified, sequenced with primer pair, ITS1/ITS4 (2), and submitted into GenBank with an accession number, MW228165. Phylogenetic analysis was conducted with sequences of reference strains (3). The result showed that the fungus obtained in this study was fallen into the same group with N.oryzae. In view of above both morphological and molecular analysis, the strains were finally identified as N. oryzae. Pathogenicity tests were conducted in triplicate with rice panicles in initial heading stage. Fifty panicles were wounded on branches with needles and inoculated by spreading the conidia suspension (10μl, 1 × 106 conidia ml-1) on the wounds. The panicles used as control were treated in same way with 10μl of sterile water. The inoculated and control plants were kept in dark, 25 ℃ and relative humidity of more than 85% for 24 h in culture chamber. Symptoms appeared on 44 of 50 inoculated panicles which were basically similar with those observed in paddy field, while negative controls remained symptomless. The fungi re-isolated from inoculated panicles were also confirmed as N. oryzae by both morphological and molecular analysis. To the best of our knowledge, this is the first report of N. oryzae causing panicle branch rot disease on Oryza sativa (rice). This disease not only cause yield losses and lower milling quality, but could also be mistaken as rice blast incurring unnecessary fungicides spray.
PubMed: 33728954
DOI: 10.1094/PDIS-11-20-2423-PDN -
Plant Disease Jun 2021In mid-August 2020, small circular tan-colored lesions were detected in leaves of corn, Zea mays, in Sussex county Delaware. Symptoms were observed in many fields,...
In mid-August 2020, small circular tan-colored lesions were detected in leaves of corn, Zea mays, in Sussex county Delaware. Symptoms were observed in many fields, affecting multiple hybrids that were at approximately the dough growth stage (R4). Lesions were present on most plants and individual ear leaf severity ratings ranged from 3 to 14 %. Symptomatic corn leaves were selected from the field and kept in a moist chamber for 24 hrs at 25 °C. In lesions, melanized spores were observed ranging from 21.94 µm to 30.51 µm in length and 7.83µm to 11.70µm in width (n=20). One cm2 leaf sections were extracted and sterilized in a 0.85% sodium hypochlorite solution for 30s followed by a sterile water rinse for 30s. Leaf pieces were then plated onto potato dextrose agar amended with 50µg/ml penicillin-G-sodium salt and streptomycin sulfate. Petri dishes were incubated at 25 °C with a 12-hr photoperiod for 14 days. Colonies were brown-black to lighter gray in color. Media was stained from orange, to dark brown, or not at all. Conidia were melanized and curved with three transverse septa matching the size above. Colony and spore morphology were consistent with the description of Curvularia lunata (Garcia-Aroca et al. 2018). Pure cultures were obtained, and a representative isolate was sequenced to confirm fungal identity by amplifying the internal transcribed spacer (ITS) region with primers ITS4 and ITS5. BLAST search results confirmed 100% similarity (483/531 bp) to the C. lunata reference sequence (MK623264). The sequence was deposited in GenBank as accession number MW794323. To complete Koch's postulates, corn (hybrid: Hubner H6187RCSS) was planted into 10 cm pots and maintained at 25°C and a 12 hr photoperiod. A conidial suspension was made by soaking a Petri dish of a 3-week-old fungal colony with 20 ml of sterile water and a drop of Tween-20 solution then, scraping mycelia to dislodge spores. The conidial concentration was calibrated to 5 x 105 spores/ml and 15 ml were applied to each whorl of four three-collar stage plants using a Preval sprayer (Nakoma Products, Bridgeview, IL). Four plants were inoculated with 15 ml sterile DI water as a control. This experiment was repeated twice. Each plant was moved to an incubator and covered with a plastic bag for 24 hr to maintain humidity. Conditions in the incubator were maintained at 25 °C with a 12-hr photoperiod. Inoculated plants displayed small, oval-shaped lesions within four days. No symptoms were observed on the control plants. Symptomatic leaves were harvested and placed into a moist chamber for 24 hrs to sporulate and lesions were plated onto PDA as described above. Culture morphology was consistent with the original isolate, with spores slightly larger ranging from 22.35 µm to 39.29 µm in length, and 8.36 µm to 11.69 µm in width (n=20). Isolates obtained from inoculated plants were sequenced and maintained 100% identity with the reference sequence described above. C. lunata was first reported in Louisiana in 2017 (Garcia-Aroca et al. 2018) and in Kentucky in 2018 (Anderson et al. 2019). This is the first report of Curvularia leaf spot on corn in Delaware. Symptoms developed late in the season, so it is unlikely that yield was affected in 2020. However, the economic impact of this disease in the United States is still unclear, it will be important to monitor potential impacts of this disease in Delaware corn production. References 1.Anderson NR, et al., 2019, Plant Disease, 103, 2692, doi: 0.1094/PDIS-03-19-0629-PDN. 2. Garcia-Aroca T, et al., 2018, Plant health progress, 19, 140, doi: 10.1094/PHP-02-18-0008-BR.
PubMed: 34170757
DOI: 10.1094/PDIS-04-21-0742-PDN -
MicroLife 2023On 9-13 July 2023, the 10th FEMS Congress took place in Hamburg, Germany. As part of this major event in European microbiology, the European Academy of Microbiology...
On 9-13 July 2023, the 10th FEMS Congress took place in Hamburg, Germany. As part of this major event in European microbiology, the European Academy of Microbiology (EAM) organized two full sessions. One of these sessions aimed to highlight the research of four recently elected EAM fellows and saw presentations on bacterial group behaviours and development of resistance to antibiotics, as well as on new RNA viruses including bacteriophages and giant viruses of amoebae. The other session included five frontline environmental microbiologists who showcased real-world examples of how human activities have disrupted the balance in microbial ecosystems, not just to assess the current situation but also to explore fresh approaches for coping with external disturbances. Both sessions were very well attended, and no doubt helped to gain the EAM and its fellows more visibility.
PubMed: 38107236
DOI: 10.1093/femsml/uqad045 -
Plants (Basel, Switzerland) Aug 2021The present study was aimed at investigating the allelopathic effects of a crude extract from (L.) R.M.King and H.Rob. (Siam weed). The effects of 70% crude ethanol...
The present study was aimed at investigating the allelopathic effects of a crude extract from (L.) R.M.King and H.Rob. (Siam weed). The effects of 70% crude ethanol extract from the whole plant, leaf, stem, and root on the germination and growth of and seedlings were evaluated using Petri-dish tests under laboratory conditions. Crude extracts from the leaf showed the highest inhibitory activity. The leaf extract (OR) was further separated by sequential solvent extraction to provide hexane (HX), ethyl acetate (ET), and butanol (BU) fractions, which were also evaluated using Petri-dish tests. The hexane fraction was significantly the most active; therefore, it was selected for formulation in a concentrated suspension and tested for its herbicidal characteristics. The formulation showed greater early post-emergence than post- and pre-emergence activities, respectively. The physiological mechanism of the formulation was tested against and showed that chlorophyll a and b and the carotenoid contents of the leaf dramatically decreased when the concentration was increased, suggesting its ability to disrupt the process of photosynthesis. As thiobarbituric acid reactive substances also occurred in the leaf of , this suggests lipid peroxidation and cell disruption. These results represent the possibility that extract contains inhibitory compounds with herbicidal activity and could be used as an early post-emergence herbicide for weed control.
PubMed: 34451653
DOI: 10.3390/plants10081609 -
Frontiers in Microbiology 2019is an ascomycete fungus that infects and contaminates corn, peanuts, cottonseed, and treenuts with acutely toxic and carcinogenic aflatoxins. The ecological function of...
is an ascomycete fungus that infects and contaminates corn, peanuts, cottonseed, and treenuts with acutely toxic and carcinogenic aflatoxins. The ecological function of aflatoxin production is not well understood; though not phytotoxic, aflatoxin may be involved in resisting oxidative stress responses from infection or drought stress in plants. Observation of aflatoxin stimulation in 48-well plates in response to increasing inoculated wells sparked an investigation to determine if volatiles influence aflatoxin production in neighboring colonies. Experiments controlling several culture conditions demonstrated a stimulation of aflatoxin production with increased well occupancy independent of pH buffer, moisture, or isolate. However, even with all wells inoculated, aflatoxin production was less in interior wells. Only one isolate stimulated aflatoxin production in a large Petri-dish format containing eight small Petri dishes with shared headspace. Other isolates consistently inhibited aflatoxin production when all eight Petri dishes were inoculated with . No contact between cultures and only shared headspace implied the fungus produced inhibitory and stimulatory gases. Adding activated charcoal between wells and dishes prevented inhibition but not stimulation indicating stimulatory and inhibitory gases are different and/or gas is inhibitory at high concentration and stimulatory at lower concentrations. Characterizing stimulatory and inhibitory effects of gases in headspace as well as the apparently opposing results in the two systems deserves further investigation. Determining how gases contribute to quorum sensing and communication could facilitate managing or using the gases in modified atmospheres during grain storage to minimize aflatoxin contamination.
PubMed: 32010096
DOI: 10.3389/fmicb.2019.03038 -
Nanomaterials (Basel, Switzerland) Jul 2022Heating has recently been applied as an alternative to electrical stimulation to modulate excitability and to induce neuritogenesis and the expression of neuronal...
Heating has recently been applied as an alternative to electrical stimulation to modulate excitability and to induce neuritogenesis and the expression of neuronal markers; however, a long-term functional differentiation has not been described so far. Here, we present the results obtained by a new approach for scalable thermal stimulation on the behavior of a model of dorsal root ganglion neurons, the F-11 cell line. Initially, we performed experiments of bulk stimulation in an incubator for different time intervals and temperatures, and significant differences in neurite elongation and in electrophysiological properties were observed in cultures exposed at 41.5 °C for 30 min. Thus, we exposed the cultures to the same temperature increase using a near-infrared laser to irradiate a disc of Prussian blue nanoparticles and poly-vinyl alcohol that we had adhered to the outer surface of the petri dish. In irradiated cells, neurites were significantly longer, and the electrophysiological properties (action potential firing frequency and spontaneous activity) were significantly increased compared to the control. These results show for the first time that a targeted thermal stimulation could induce morphological and functional neuronal differentiation and support the future application of this method as a strategy to modify neuronal behavior in vivo.
PubMed: 35808140
DOI: 10.3390/nano12132304