scholarly journals Hybrid Genotype of Anisakis simplex (s.s.) and A. pegreffii Identified in Third- and Fourth-Stage Larvae from Sympatric and Allopatric Spanish Marine Waters

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2458
Author(s):  
Xavier Roca-Geronès ◽  
M. Magdalena Alcover ◽  
Carla Godínez-González ◽  
Isabel Montoliu ◽  
Roser Fisa
Keyword(s):  
Marine Mammals ◽  
Rflp Analysis ◽  
Its Region ◽  
Anisakis Simplex ◽  
Fourth Stage ◽  
Growth Behaviour ◽  
Marine Waters ◽  
Hybrid Genotype ◽  
North East

The sibling species Anisakis simplex (s.s.) and Anisakis pegreffii are parasites of marine mammals and fish worldwide and the main causative agents of human anisakiasis. In sympatric areas, a hybrid genotype between the two species has been identified, mainly in third-stage larvae, but rarely in fourth-stage and adult forms. The aim of this study was to confirm the presence of hybrid genotypes in larvae parasitizing fish caught in sympatric and allopatric Spanish marine waters, the North-East Atlantic and West Mediterranean, respectively, and to study possible differences in the growth behaviour between genotypes. Of the 254 molecularly analysed larvae, 18 were identified as hybrids by PCR-RFLP analysis of the rDNA ITS region, 11 of which were subsequently confirmed by EF1 α-1 nDNA gene sequencing. These results therefore indicate an overestimation of hybrid genotypes when identification is based only on the ITS region. We also report the detection of a hybrid specimen in a host from the West Mediterranean, considered an allopatric zone. Additionally, fourth-stage larvae with a hybrid genotype were obtained in vitro for the first time, and no differences were observed in their growth behaviour compared to larvae with A. simplex (s.s.) and A. pegreffii genotypes.

scholarly journals Correlation of NHR-48 Transcriptional Modulator Expression with Selected CYP Genes’ Expression during Thiabendazole Treatment of Anisakis simplex (s.l.)?—An In Vitro Study

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1030
Author(s):  
Elżbieta Łopieńska-Biernat ◽  
Robert Stryiński ◽  
Łukasz Paukszto ◽  
Jan P. Jastrzębski
Keyword(s):  
Marine Mammals ◽  
Hormone Receptors ◽  
Wide Spectrum ◽  
Cytochromes P450 ◽  
In Vitro Study ◽  
Biological Species ◽  
Anisakis Simplex ◽  
Complex Life Cycle ◽  
Parasitic Nematodes

Anisakis simplex (s.l.) is a complex of three sibling (biological) species of parasitic nematodes of marine mammals, including A. berlandi, A. pegreffii and A. simplex (s.s.). It is characterized by a complex life cycle in which humans can become accidental hosts by consuming dishes made of raw or undercooked fish containing L3 larvae, which in many regions of the world is related to the national or regional culinary tradition. This has spurred scientific efforts to develop new methods for treating the disease, called anisakiasis, and to neutralize invasive L3. Thiabendazole (TBZ) is a wide-spectrum anthelminthic with a higher efficacy than albendazole, a drug whose long-term use induces resistance in many parasitic species. Cytochromes P450 participate in TBZ metabolism, and the expression of their genes is controlled by nuclear hormone receptors (NHR). This study aimed to examine the effects of TBZ on the above-described pathway in invasive larvae of A. simplex (s.l.). The efficacy of TBZ against A. simplex (s.l.) larvae was observed for the first time. Larvae were cultured in vitro for 72 h in a medium containing TBZ at five concentrations from 0.5 to 1.5 mM. However, the survival curves did not significantly differ from each other. This means that all of the concentrations of TBZ had a similar effect on the A. simplex (s.l.) L3 larvae during in vitro culture. Nevertheless, TBZ modified the expression of nhr-48, cyp13a3 and cyp1a1 genes in the L3 of A. simplex (s.l.).

scholarly journals Angiostrongylus cantonensis (Nematode: Metastrongiloidea): In Vitro Cultivation of Infective Third-Stage Larvae to Fourth-Stage Larvae

PLoS ONE ◽  
2013 ◽  
Vol 8 (8) ◽  
pp. e72084 ◽  
Author(s):  
Rong-Jyh Lin ◽  
Jie-Wen He ◽  
Li-Yu Chung ◽  
June-Der Lee ◽  
Jiun-Jye Wang ◽  
...  
Keyword(s):  
Angiostrongylus Cantonensis ◽  
In Vitro Cultivation ◽  
Fourth Stage ◽  
Third Stage

scholarly journals Two new species of Undifilum, fungal endophytes of Astragalus (locoweeds) in the United States

Botany ◽  
2012 ◽  
Vol 90 (9) ◽  
pp. 866-875 ◽  
Author(s):  
Deana L. Baucom ◽  
Marie Romero ◽  
Robert Belfon ◽  
Rebecca Creamer
Keyword(s):  
New Species ◽  
New Mexico ◽  
Random Amplified Polymorphic Dna ◽  
Phylogenetic Analyses ◽  
Fungal Endophytes ◽  
Morphological Characteristics ◽  
Its Region ◽  
The United States ◽  
Neighbor Joining

New species of Undifilum , from locoweeds Astragalus lentiginosus Vitman and Astragalus mollissimus Torr., are described using morphological characteristics and molecular phylogenetic analyses as Undifilum fulvum Baucom & Creamer sp. nov. and Undifilum cinereum Baucom & Creamer sp. nov. Fungi were isolated from dried plants of A. lentiginosus var. araneosus , diphysus , lentiginosus , and wahweapensis collected from Arizona, Oregon, and Utah, USA, and A. mollissimus var. biglovii , earleii , and mollissimus collected from New Mexico, Oklahoma, and Texas, USA. Endophytic fungi from Astragalus locoweeds were compared to Undifilum oxytropis isolates obtained from dried plant material of Oxytropis lamberteii from New Mexico and Oxytropis sericea from Arizona, Colorado, New Mexico, Utah, and Wyoming. Extremely slow growth in vitro was observed for all, and conidia, if present, were ellipsoid with transverse septa. However, in vitro color, growth on four different media, and conidium size differed between fungi from Astragalus spp. and U. oxytropis. Neighbor-joining analyses of internal transcribed spacer (ITS) region and glyceraldehyde-3-phosphate dehydrogenase (GPD) gene sequences revealed that U. fulvum and U. cinereum formed a clade distinct from U. oxytropis. This was supported by neighbor-joining analyses of results generated from random amplified polymorphic DNA (RAPD) fragments using two different primers.

scholarly journals New Isolated Metschnikowia pulcherrima Strains from Apples for Postharvest Biocontrol of Penicillium expansum and Patulin Accumulation

Toxins ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 397
Author(s):  
Laura Settier-Ramírez ◽  
Gracia López-Carballo ◽  
Pilar Hernández-Muñoz ◽  
Angélique Fontana ◽  
Caroline Strub ◽  
...  
Keyword(s):  
Liquid Medium ◽  
Fungal Growth ◽  
Its Region ◽  
Antagonistic Activity ◽  
Penicillium Expansum ◽  
Metschnikowia Pulcherrima ◽  
Potential Activity ◽  
Different Strains ◽  
Main Producer

Wild yeasts isolated from the surface of apples were screened for antagonistic activity against Penicillium expansum, the main producer of the mycotoxin patulin. Three antagonistic yeasts (Y33, Y29 and Y24) from a total of 90 were found to inhibit P. expansum growth. Identification by ITS region sequence and characterization showed that three selected isolates of yeast should be different strains of Metschnikowia pulcherrima. Several concentrations of the selected yeasts were used to study their in vitro antifungal effectivity against P. expansum on Petri dishes (plates with 63.6 cm2 surface) whereas their potential activity on patulin reduction was studied in liquid medium. Finally, the BCA that had the best in vitro antifungal capacity against P. and the best patulin degradation capacity was selected to be assessed directly on apples. All the selected strains demonstrated antifungal activity in vitro but the most efficient was the strain Y29. Isolated strains were able to reduce patulin content in liquid medium, Y29 being the only strain that completely reduced patulin levels within 120 h. The application of Y29 as biocontrol agent on the surface of apples inoculated with P. expansum, inhibited fungal growth and patulin production during storage. Therefore, the results shown that this yeast strain could be used for the reduction of P. expansum and its mycotoxin in apples or apple-based products by adapting the procedure application.

Radiation activates the NORAD expression to promote ESCC radioimmunotherapy resistance via EEPD1/ATR/Chk1 signaling by inhibiting the pri-miR-199 processing and exosomal transfer of miR-199a-5p

2021 ◽  
Author(s):  
Yuchen Sun ◽  
Jizhao Wang ◽  
Xuanzi Sun ◽  
Jing Li ◽  
Xu Zhao ◽  
...  
Keyword(s):  
Its Region ◽  
Luciferase Reporter ◽  
In Vivo Experiments ◽  
Cycle Arrest ◽  
Non Coding Rna ◽  
Irradiation Treatment ◽  
Recombination Repair ◽  
Reporter Assays

Abstract Background Radioresistance, a poorly understood phenomenon, results in the failure of radiotherapy and consequent local recurrence, threatening a large proportion of ESCC patients. To date, lncRNAs have been found to be involved in diverse biological processes, including radioresistance.Methods ELISA was used to evaluated the H3 modifications in radio-resistant ESCC cells. FISH and qRT-PCR were adopted to examine the expression and localization of lncRNA-NORAD, pri-miR-199a and miR-199a. Electron microscopy and Nanoparticle tracking analysis (NTA) was conducted to observe and identify exosomes. High-throughput RNA sequencing and TMT mass spectrometry were performed to identify the functional lncRNAs and proteins involved in ESCC radioresistance. A series of in vitro and in vivo experiments were performed to investigate the biological effect of NORAD. CHIP, qPCR-RIP, co-IP and dual-luciferase reporter assays were used to explore the interaction of related RNAs and proteins. Results We show here that a DNA damage activated non-coding RNA-NORAD, which is critical for ESCC radio-resistance. NORAD was highly expressed in radio-resistant ESCC cells and tissues. Irradiation treatment promotes NORAD expression via enhancing H3K4me2 enrichment on its region. NORAD knockdown cells exhibit significantly hypersensitivity to irradiation in vivo and in vitro. NORAD is required for initiating repair and restart of stalled forks, G2 cycle arrest and homologous recombination repair upon irradiation treatment. Mechanistically, NORAD inhibits miR-199a expression by competitively binding PUM1 from pri-miR-199a, inhibiting the process of pri-miR-199a. Mature miR-199a in NORAD-knockdown cells can be packaged into exosomes; miR-199a restores the radiosensitivity of radioresistant cells by targeting EEPD1, then inhibiting ATR/Chk1 signaling pathway. Simultaneously, NORAD knockdown blocks the ubiquitination of PD-L1, leads to the better response for radiation and anti-PD-1 treatment in mouse model.Conclusion This study raises the possibility that LncRNA-NORAD could be a potential treatment target for improving the efficiency of immunotherapy in combination with radiation in ESCC.

Cytosolic glutathione S-transferases of Oesophagostomum dentatum

Parasitology ◽  
2008 ◽  
Vol 135 (10) ◽  
pp. 1215-1223 ◽  
Author(s):  
A. JOACHIM ◽  
B. RUTTKOWSKI
Keyword(s):  
Amino Acids ◽  
Pcr Primers ◽  
Mrna Levels ◽  
Fourth Stage ◽  
Glutathione S Transferase ◽  
Cdna Sequences ◽  
Oesophagostomum Dentatum ◽  
Cytosolic Glutathione ◽  
Glutathione S Transferases

SUMMARYOesophagostomum dentatum stages were investigated for glutathione S-transferase (GST) expression at the protein and mRNA levels. GST activity was detected in all stages (infectious and parasitic stages including third- and fourth-stage larvae of different ages as well as males and females) and could be dose-dependently inhibited with sulfobromophthalein (SBP). Addition of SBP to in vitro larval cultures reversibly inhibited development from third- to fourth-stage larvae. Two glutathione-affinity purified proteins (23 and 25 kDa) were detected in lysates of exsheathed third-stage larvae by SDS-PAGE. PCR-primers were designed based on peptide sequences and conserved GST sequences of other nematodes for complete cDNA sequences (621 and 624 nt) of 2 isoforms, Od-GST1 and Od-GST2, with 72% nucleotide similarity and 75% for the deduced proteins. Genomic sequences consisted of 7 exons and 6 introns spanning 1296 bp for Od-GST1 and 1579 and 1606 bp for Od-GST2. Quantitative real-time-PCR revealed considerably elevated levels of Od-GST1 in the early parasitic stages and slightly reduced levels of Od-GST2 in male worms. Both Od-GSTs were most similar to GST of Ancylostoma caninum (nucleotides: 73 and 70%; amino acids: 80 and 73%). The first three exons (75 amino acids) corresponded to a synthetic prostaglandin D2 synthase (53% similarity). O. dentatum GSTs might be involved in intrinsic metabolic pathways which could play a role both in nematode physiology and in host-parasite interactions.

scholarly journals Antifungal activity of several isolates of Trichoderma against Cladosporium and Botrytis

2018 ◽  
Vol 8 (1) ◽  
pp. 88-91 ◽  
Author(s):  
M. Skaptsov ◽  
S. Smirnov ◽  
M. Kutsev ◽  
O. Uvarova ◽  
T. Sinitsyna ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Trichoderma Harzianum ◽  
Its Region ◽  
Botanical Garden ◽  
Antagonistic Activity ◽  
Antagonistic Effect ◽  
Soil Samples ◽  
Morphological Observation ◽  
The South

<p><em>Trichoderma</em> isolates (SSBGT07, SSBGT08, SSBGT09, SSBGT10) were isolated from the soil samples of the South-Siberian Botanical Garden and identified using morphological observation and ITS region analysis as <em>Trichoderma harzianum</em>, <em>T. asperellum, T. ghanense</em>, and <em>T. longibranchiatum</em>. Antagonistic activity against <em>Cladosporium </em>sp. and<em> Botrytis </em>sp. was evaluated <em>in vitro</em>. All isolates showed antagonistic effect by competition against <em>Cladosporium </em>sp. <em>T. asperellum </em>and <em>T. longibranchiatum</em> showed antagonism against <em>Botrytis </em>sp. All isolates showed hyper sporulation on the sclerotia of <em>Botrytis</em> sp. (except the <em>T. ghanense</em>) and colonies of the <em>Cladosporium</em> sp. Our study provides new isolates that affect the <em>Cladosporium </em>sp. and<em> Botrytis </em>sp.</p>

scholarly journals Effective Management of Cucumber Powdery Mildew with Essential Oils

Agriculture ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1177
Author(s):  
Yasser S. Mostafa ◽  
Mohamed Hashem ◽  
Ali M. Alshehri ◽  
Saad Alamri ◽  
Ebrahem M. Eid ◽  
...  
Keyword(s):  
Powdery Mildew ◽  
Essential Oils ◽  
Disease Incidence ◽  
Its Region ◽  
Dry Weight ◽  
Positive Control ◽  
Powdery Mildew Fungus ◽  
Membrane Injury ◽  
Cucumber Powdery Mildew

This research evaluated the efficacy of essential oils in the management of cucumber powdery mildew. Essential oils of lemongrass, lemon, thyme, peppermint, abundance blend, purification blend, and thieves blend were tested in vitro and under greenhouse conditions in two separate experiments. The effects of essential oils were tested against powdery mildew disease at concentrations of 1.0–2.5 mL/L, and the consequent impact of the oils on plant growth was evaluated. Powdery mildew fungus, Podosphaera xanthii, was identified using sequencing of the ITS region. The essential oils significantly reduced disease incidence up to 77.3% compared with the positive control (p < 0.5). Moreover, the essential oils increased the plant length (up to 187 cm), leaf area (up to 27.5 cm2), fresh weight (up to 123 g), dry weight (up to 22.5 g), number of flowers (16.3), and metabolite content compared with the positive control (p < 0.5). Cell membrane injury decreased significantly in the oil-treated pants (p < 0.5), indicating the protective effect of essential oils. This study recommends the application of essential oils in an appropriate dose (2.5 mL/L) to protect cucumber plants against powdery mildew. Overdose of the oils (more than 2.5 mL/L) should be avoided due to adverse effects.

scholarly journals Inhibitory Effect of Trichoderma asperellum Isolate against Ralstonia solanacearum Causing Brinjal Wilt

2021 ◽  
Vol 24 (2) ◽  
pp. 107-120
Author(s):  
SMN Islam ◽  
SS Siddique ◽  
MZH Chowdhury ◽  
NJ Mishu
Keyword(s):  
Ralstonia Solanacearum ◽  
Disease Incidence ◽  
Its Region ◽  
Inhibitory Effect ◽  
Dual Culture ◽  
Trichoderma Asperellum ◽  
In Planta ◽  
Quantitative Test ◽  
Trichoderma Isolate

A native Trichoderma isolate was collected from the agricultural soil of Gazipur. This isolate was identified as a Trichoderma asperellum through morphology and analysis of internal transcribed spacer (ITS) region of ribosomal RNA gene sequence and reconstruction of the phylogenetic tree. The antagonistic effects of the newly identified T. asperellum isolate were assessed against brinjal bacterial wilt caused by Ralstonia solanacearum both in vitro and in planta. Both qualitative and quantitative bioassays were conducted in vitro. For qualitative tests, dual culture and antibacterial activity were carried out, and pathogen growth was observed visually. The antagonism of T. asperellum cell free culture filtrate on the growth of R. solanacearum was conducted in a quantitative test. Successful antagonism was recorded after both in vitro qualitative tests. In addition, the lowest colony forming unit was recorded in 100% of CFC (2.4±0.51 ×103 cfu/ml) in quantitative test. The T. asperellum inoculated plant showed low disease incidence (13.33%) when seedlings were challenged with R. solanacearum in planta experiment. Disease incidence was 100% for seedlings when treated with only R. solanacearum. The results showed that the isolated and identified T. asperellum isolate suppressed R. solanacearum growth in vitro and protected the seedling from wilting in planta. Therefore, this isolate could be considered as a potential isolate. Ann. Bangladesh Agric. (2020) 24(2) : 107-120

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