Detection of Phytophthora erythroseptica in Soil Using Nightshade as Bait Combined with PCR Techniques

Enteric viruses are the leading cause of diarrhea in children globally. Identifying viral agents and understanding their genetic diversity could help to develop effective preventive measures. This study aimed to determine the detection rate and genetic diversity of four enteric viruses in Gabonese children aged below five years. Stool samples from children <5 years with (n = 177) and without (n = 67) diarrhea were collected from April 2018 to November 2019. Norovirus, astrovirus, sapovirus, and aichivirus A were identified using PCR techniques followed by sequencing and phylogenetic analyses. At least one viral agent was identified in 23.2% and 14.9% of the symptomatic and asymptomatic participants, respectively. Norovirus (14.7%) and astrovirus (7.3%) were the most prevalent in children with diarrhea, whereas in the healthy group norovirus (9%) followed by the first reported aichivirus A in Gabon (6%) were predominant. The predominant norovirus genogroup was GII, consisting mostly of genotype GII.P31-GII.4 Sydney. Phylogenetic analysis of the 3CD region of the aichivirus A genome revealed the presence of two genotypes (A and C) in the study cohort. Astrovirus and sapovirus showed a high diversity, with five different astrovirus genotypes and four sapovirus genotypes, respectively. Our findings give new insights into the circulation and genetic diversity of enteric viruses in Gabonese children.

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Nutrient Deficiency Promotes the Entry of Helicobacter pylori Cells into Candida Yeast Cells

Biology ◽

10.3390/biology10050426 ◽

2021 ◽

Vol 10 (5) ◽

pp. 426

Author(s):

Kimberly Sánchez-Alonzo ◽

Fabiola Silva-Mieres ◽

Luciano Arellano-Arriagada ◽

Cristian Parra-Sepúlveda ◽

Humberto Bernasconi ◽

...

Keyword(s):

Helicobacter Pylori ◽

Nutrient Deficiency ◽

Gastric Epithelium ◽

Yeast Cells ◽

Nutrient Concentrations ◽

Fetal Bovine ◽

H Pylori ◽

Pcr Techniques ◽

Strain Dependent

Helicobacter pylori, a Gram-negative bacterium, has as a natural niche the human gastric epithelium. This pathogen has been reported to enter into Candida yeast cells; however, factors triggering this endosymbiotic relationship remain unknown. The aim of this work was to evaluate in vitro if variations in nutrient concentration in the cultured medium trigger the internalization of H. pylori within Candida cells. We used H. pylori–Candida co-cultures in Brucella broth supplemented with 1%, 5% or 20% fetal bovine serum or in saline solution. Intra-yeast bacteria-like bodies (BLBs) were observed using optical microscopy, while intra-yeast BLBs were identified as H. pylori using FISH and PCR techniques. Intra-yeast H. pylori (BLBs) viability was confirmed using the LIVE/DEAD BacLight Bacterial Viability kit. Intra-yeast H. pylori was present in all combinations of bacteria–yeast strains co-cultured. However, the percentages of yeast cells harboring bacteria (Y-BLBs) varied according to nutrient concentrations and also were strain-dependent. In conclusion, reduced nutrients stresses H. pylori, promoting its entry into Candida cells. The starvation of both H. pylori and Candida strains reduced the percentages of Y-BLBs, suggesting that starving yeast cells may be less capable of harboring stressed H. pylori cells. Moreover, the endosymbiotic relationship between H. pylori and Candida is dependent on the strains co-cultured.

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Characterization of Ocular Surface Microbial Profiles Revealed Discrepancies between Conjunctival and Corneal Microbiota

Pathogens ◽

10.3390/pathogens10040405 ◽

2021 ◽

Vol 10 (4) ◽

pp. 405

Author(s):

Anna Matysiak ◽

Michal Kabza ◽

Justyna A. Karolak ◽

Marcelina M. Jaworska ◽

Malgorzata Rydzanicz ◽

...

Keyword(s):

Microbial Communities ◽

Ocular Surface ◽

Molecular Techniques ◽

Corneal Tissue ◽

Rna Seq ◽

Microbiome Composition ◽

Viral Sequences ◽

Pcr Techniques ◽

Unmapped Reads

The ocular microbiome composition has only been partially characterized. Here, we used RNA-sequencing (RNA-Seq) data to assess microbial diversity in human corneal tissue. Additionally, conjunctival swab samples were examined to characterize ocular surface microbiota. Short RNA-Seq reads, obtained from a previous transcriptome study of 50 corneal tissues, were mapped to the human reference genome GRCh38 to remove sequences of human origin. The unmapped reads were then used for taxonomic classification by comparing them with known bacterial, archaeal, and viral sequences from public databases. The components of microbial communities were identified and characterized using both conventional microbiology and polymerase chain reaction (PCR) techniques in 36 conjunctival swabs. The majority of ocular samples examined by conventional and molecular techniques showed very similar microbial taxonomic profiles, with most of the microorganisms being classified into Proteobacteria, Firmicutes, and Actinobacteria phyla. Only 50% of conjunctival samples exhibited bacterial growth. The PCR detection provided a broader overview of positive results for conjunctival materials. The RNA-Seq assessment revealed significant variability of the corneal microbial communities, including fastidious bacteria and viruses. The use of the combined techniques allowed for a comprehensive characterization of the eye microbiome’s elements, especially in aspects of microbiota diversity.

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In-Furrow Applications of Metalaxyl and Phosphite for Control of Pink Rot (Phytophthora erythroseptica) of Potato in New Brunswick, Canada

Plant Disease ◽

10.1094/pdis-91-10-1305 ◽

2007 ◽

Vol 91 (10) ◽

pp. 1305-1309 ◽

Cited By ~ 12

Author(s):

Khalil I. Al-Mughrabi ◽

Rick D. Peters ◽

H. W. (Bud) Platt ◽

Gilles Moreau ◽

Appanna Vikram ◽

...

Keyword(s):

New Brunswick ◽

Disease Incidence ◽

Field Trials ◽

Russet Burbank ◽

Viable Option ◽

Potato Plants ◽

Pink Rot ◽

Phytophthora Erythroseptica ◽

Sensitive Isolate ◽

The Mean

The efficacy of metalaxyl-m (Ridomil Gold 480EC) and phosphite (Phostrol) applied at planting in-furrow against pink rot (Phytophthora erythroseptica) of potato (Solanum tuberosum) ‘Shepody’ and ‘Russet Burbank’ was evaluated in field trials conducted in 2005 and 2006 in Florenceville, New Brunswick, Canada. Inoculum made from a metalaxyl-m-sensitive isolate of P. erythroseptica from New Brunswick was applied either in-furrow as a vermiculite slurry at planting or as a zoospore drench in soils adjacent to potato plants in late August. After harvest, the number and weight of tubers showing pink rot symptoms were assessed and expressed as percentages of the total tuber number and total weight of tubers. Metalaxyl-m applied in-furrow was significantly more effective against pink rot than phosphite. The mean percentage of diseased tubers as a percentage of total tuber weight was 1.5% (2005) and 1.2% (2006) for metalaxyl-m-treated plots and 9.6% (2005) and 2.8% (2006) for phosphite-treated plots, a percentage similar to that obtained in inoculated control plots with no fungicide treatment. The mean percentage of diseased tubers expressed as a percentage of the total number of tubers was 1.7% (2005) and 1.3% (2006) for metalaxyl-m-treated plots and 10.1% (2005) and 3.1% (2006) for phosphite-treated plots. Disease incidence was significantly higher using the late-season inoculation technique (respective means in 2005 and 2006 were 9.9 and 3.8% diseased tubers, by weight, and 10.6 and 3.9%, by number) than with the in-furrow inoculation method (respective means in 2005 and 2006 were 3.3 and 0.7% by weight, and 3.7 and 1.3%, by number). The potato cv. Shepody was significantly more susceptible to pink rot (9.9 and 3.3% diseased tubers, by weight, in 2005 and 2006, respectively, and 10.6 and 3.9%, by number) than Russet Burbank (respective means in 2005 and 2006 were 3.4,% and 1.2%, by weight, and 3.7,% and 1.2%, by number). Our findings indicate that metalaxyl applied in-furrow at planting is a viable option for control of pink rot caused by metalaxyl-sensitive strains of P. erythroseptica, whereas phosphite was ineffective.

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Limited Genetic Diversity in North American Isolates of Phytophthora erythroseptica Pathogenic to Potato Based on RAPD Analysis

Plant Disease ◽

10.1094/pd-89-0380 ◽

2005 ◽

Vol 89 (4) ◽

pp. 380-384 ◽

Cited By ~ 10

Author(s):

Rick D. Peters ◽

Rod J. Clark ◽

Albert D. Coffin ◽

Antony V. Sturz ◽

David H. Lambert ◽

...

Keyword(s):

Genetic Diversity ◽

North America ◽

New Brunswick ◽

Prince Edward Island ◽

Random Amplified Polymorphic Dna ◽

Geographic Origin ◽

The United States ◽

Vitro Assay ◽

Phytophthora Erythroseptica

Pink rot of potato (Solanum tuberosum), caused by Phytophthora erythroseptica, is found wherever potatoes are grown, and in the last decade, it has reemerged as an economically important disease in Canada and the United States. A selection of isolates of P. erythroseptica from major potato-growing regions in North America, namely Prince Edward Island and New Brunswick, Canada, and Maine and Idaho, U.S.A., was assessed for genetic diversity with randomly chosen decanucleotide primers which were used to amplify regions of DNA to reveal polymorphisms among templates (random amplified polymorphic DNA [RAPD]). The isolates varied in their geographic origin as well as in their sensitivity to mefenoxam, as determined by an in vitro assay. In three separate RAPD screens (I, II, and III) with 23 isolates of P. erythroseptica chosen from a larger collection, 1,410, 369, and 316 robust, scorable bands were amplified, respectively. However, among the bands amplified in screens I, II, and III, only 3, 1, and 3 bands, respectively, were polymorphic. When three primers yielding polymorphisms were used to screen 106 isolates from Prince Edward Island and New Brunswick, or a representative collection of 32 isolates from Prince Edward Island, New Brunswick, Maine, and Idaho, no major variation was discovered. RAPD markers were not correlated with geographic origin or mefenoxam sensitivity of the isolates. From an evolutionary standpoint, the absence of genetic diversity among the isolates of P. erythroseptica we examined may be attributable to the relatively recent introduction of a small founding population of the pathogen in North America.

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The Progesterone Receptor in Human Term Amniochorion and Placenta Is Isoform C

Endocrinology ◽

10.1210/en.2005-0510 ◽

2006 ◽

Vol 147 (2) ◽

pp. 687-693 ◽

Cited By ~ 20

Author(s):

Anthony H. Taylor ◽

Penny C. McParland ◽

David J. Taylor ◽

Stephen C. Bell

Keyword(s):

Progesterone Receptor ◽

Western Blotting ◽

Cell Types ◽

Specific Role ◽

Fetal Membranes ◽

Rt Pcr ◽

Reproductive Tissues ◽

Pcr Techniques ◽

Human Parturition ◽

Extraembryonic Tissues

The mechanism that initiates human parturition has been proposed to be functional progesterone withdrawal whereby the 116-kDa B isoform of the progesterone receptor (PR-B) switches in favor of the 94-kDa A isoform (PR-A) in reproductive tissues. Recently other PR isoforms, PR-S, PR-C, and PR-M generated from the same gene have been identified and partially characterized. Using immunohistochemical, Western blotting, and RT-PCR techniques, evidence is provided that the major PR isoform present in human term fetal membranes (amnion and chorion) and syncytiotrophoblast of the placenta is neither of the classical nuclear PR-B or PR-A isoforms but is the N terminally truncated 60-kDa PR-C isoform. Evidence is also provided that the PR-C isoform resides in the cytoplasm of the expressing cell types. Data are also presented to show that PR-B, PR-A, and PR-S isoforms are essentially absent from the amnion and chorion, whereas PR isoforms A, B, C, and S are all present in the decidua, with PR-A being the major isoform. The syncytiotrophoblast of the placenta contains the cytoplasmic PR-C isoform but not PR-A, PR-B, or PR-S. The major PR isoform in the amnion, chorion, and placenta is PR-C, suggesting that the cytoplasmic PR-C isoform has a specific role in extraembryonic tissues and may be involved in the regulation of human parturition.

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Expression of β-catenin in human trophoblast and its role in placenta accreta and placenta previa

Journal of International Medical Research ◽

10.1177/0300060518799265 ◽

2018 ◽

Vol 47 (1) ◽

pp. 206-214

Author(s):

Qing Han ◽

Lianghui Zheng ◽

Zhaodong Liu ◽

Jinying Luo ◽

Rongxin Chen ◽

...

Keyword(s):

Western Blotting ◽

Cell Invasion ◽

Placenta Accreta ◽

Placenta Previa ◽

Control Group ◽

Rt Pcr ◽

Lesion Area ◽

Human Trophoblast ◽

Chorionic Villi ◽

Pcr Techniques

Objectives To investigate the expression of β-catenin in chorionic villi, and to explore its roles in placenta accreta and placenta previa. Methods We compared β-catenin expression in the control group, placenta accreta group (lesion area and normal zones), and placenta previa group (placental central and placental edge zones) by immunohistochemistry, Western blotting, and RT-PCR techniques. Results Compared with the normal group, the placenta accreta group had a longer length of stay, greater bleeding volume, and lower newborn birth weight. Further, the expression of β-catenin was lower in both placenta previa and placenta accreta groups than in the control group, as measured by immunohistochemistry. Compared with the control group, expression of β-catenin was significantly lower in the placenta previa and placenta accreta groups by Western blotting and RT-PCR. Importantly, the level of placental β-catenin was significantly different when compared between the lesion and normal zones of placenta. Conclusion The expression of β-catenin in placenta accreta might play an important role in the regulation of placental cell invasion; low expression of β-catenin in placenta accreta might be responsible for excessive trophoblastic invasion.

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Use of immunodot blot and multiplex reverse transcriptase-polymerase chain reaction in dengue virus detection in macerates of Aedes aegypti larvae

Microbiology Research ◽

10.4081/mr.2012.e13 ◽

2012 ◽

Vol 3 (1) ◽

pp. 13

Author(s):

Aline T.A. Chagas ◽

Michelle D. Oliveira ◽

Jose M.S. Mezencio ◽

Eduardo A.M. Silva ◽

Leandro L. Oliveira ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Aedes Aegypti ◽

Dengue Virus ◽

Reverse Transcriptase ◽

Epidemiological Surveillance ◽

Rt Pcr ◽

Chain Reaction ◽

Denv Serotypes ◽

Polymerase Chain ◽

Pcr Techniques

The <em>Dengue virus</em> is the main arbovirus that affects man in terms of morbidity and mortality. The detection of the virus is very important for epidemiological surveillance, so here we propose to standardize and compare the immunodot blot (IDB) and multiplex reverse transcriptase-polymerase chain reaction (M-RT-PCR) techniques to detect and characterize the dengue virus (DENV) serotypes in samples of <em>Aedes aegypti</em> larvae. Thus, the IDB and M-RT-PCR techniques were standardized using macerated samples of larvae collected in nature. The use of monoclonal antibodies in IDB has not shown great results, but DENV detection through this method was possible using polyclonal antibodies. The distinction of serotypes 1, 2 and 3 was carried out by M-RT-PCR.

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Comparison and Utilization of Repetitive-Element PCR Techniques for Typing Lactobacillus Isolates from the Chicken Gastrointestinal Tract

Applied and Environmental Microbiology ◽

10.1128/aem.01150-09 ◽

2009 ◽

Vol 75 (21) ◽

pp. 6764-6776 ◽

Cited By ~ 26

Author(s):

David P. Stephenson ◽

Robert J. Moore ◽

Gwen E. Allison

Keyword(s):

Gastrointestinal Tract ◽

Lactobacillus Reuteri ◽

Repetitive Element ◽

Necrotic Enteritis ◽

High Throughput Analysis ◽

Strain Diversity ◽

Disease Induction ◽

First Time ◽

Pcr Techniques ◽

Type Strains

ABSTRACT Three repetitive-element PCR techniques were evaluated for the ability to type strains of Lactobacillus species commonly identified in the chicken gastrointestinal tract. Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) produced species- and strain-specific profiles for Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus johnsonii, and Lactobacillus reuteri isolates. The technique typed strains within these species equally as well as pulsed-field gel electrophoresis. DNA concentration and quality did not affect the ERIC-PCR profiles, indicating that this method, unlike other high-resolution methods, can be adapted to high-throughput analysis of isolates. Subsequently, ERIC-PCR was used to type Lactobacillus species diversity of a large collection of isolates derived from chickens grown under commercial and necrotic enteritis disease induction conditions. This study has illustrated, for the first time, that there is great strain diversity within each Lactobacillus species present and has revealed that chickens raised under commercial conditions harbor greater species and strain diversity than chickens raised under necrotic enteritis disease induction conditions.

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