scholarly journals Differential Survivability of Two Genetically Similar Salmonella Thompson Strains on Pre-harvest Sweet Basil (Ocimum basilicum) Leaves

2021 ◽  
Vol 12 ◽  
Author(s):  
Ye Htut Zwe ◽  
Michelle Mei Zhen Ten ◽  
Xinyi Pang ◽  
Chun Hong Wong ◽  
Dan Li
Keyword(s):  
Gene Expression ◽  
Fresh Produce ◽  
Ocimum Basilicum ◽  
Survival Strategies ◽  
Post Inoculation ◽  
Human Pathogens ◽  
Adaxial Surface ◽  
Macroscopic Lesion ◽  
Better Than ◽  
Expression Polymorphism

Although conventionally considered an animal pathogen, recent evidence increasingly suggests that fresh produce may act as significant transmission vehicles and alternative hosts to Salmonella. This study reports the differential survivability of two genetically similar Salmonella Thompson strains (ST 889B and ST 688C) on the adaxial surface of pre-harvest basil (Ocimum basilicum) leaves. Upon inoculation, two distinct phenomena, a dried water-print or a macroscopic lesion, were observed within 24 h. ST 889B survived better than ST 688C on healthy-looking leaves without lesions, possibly due to its higher biofilm-forming ability. Both strains survived better on the leaves with lesions than on the healthy-looking leaves (ST 688C: 4.39 ± 0.68 vs. 2.18 ± 0.29; ST 889B: 4.78 ± 0.12 vs. 2.83 ± 0.18 log CFU per sample at 6 days post-inoculation). ST 889B caused the formation of lesions at a higher frequency [70/117 leaves (59.8%)] than ST 688C [35/96 leaves (36.5%)]. Thus, we highlighted two distinct Salmonella survival strategies in the basil pathosystem and demonstrated gene expression polymorphism (variations in the expression of the same set of genes) as an indispensable strategy in the colonization of plants as hosts by the human pathogens.

scholarly journals PSIV-11 Effects of very low-dose antibiotics on gene expression profiles in ileal mucosa of weaned pigs infected with a pathogenic E. coli

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 286-286
Author(s):  
Kwangwook Kim ◽  
Sungbong Jang ◽  
Yanhong Liu
Keyword(s):  
Gene Expression ◽  
Systemic Inflammation ◽  
Low Dose ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Post Inoculation ◽  
Growth Promoter ◽  
Weaned Pigs ◽  
Ileal Mucosa ◽  
E Coli

Abstract Our previous studies have shown that supplementation of low-dose antibiotic growth promoter (AGP) exacerbated growth performance and systemic inflammation of weaned pigs infected with pathogenic Escherichia coli (E. coli). The objective of this experiment, which is extension of our previous report, was to investigate the effect of low-dose AGP on gene expression in ileal mucosa of weaned pigs experimentally infected with F18 E. coli. Thirty-four pigs (6.88 ± 1.03 kg BW) were individually housed in disease containment rooms and randomly allotted to one of three treatments (9 to 13 pigs/treatment). The three dietary treatments were control diet (control), and 2 additional diets supplemented with 0.5 or 50 mg/kg of AGP (carbadox), respectively. The experiment lasted 18 d [7 d before and 11 d after first inoculation (d 0)]. The F18 E. coli inoculum was orally provided to all pigs with the dose of 1010 cfu/3 mL for 3 consecutive days. Total RNA [4 to 6 pigs/treatment on d 5; 5 to 7 pigs/treatment on 11 post-inoculation (PI)] was extracted from ileal mucosa to analyze gene expression profiles by Batch-Tag-Seq. The modulated differential gene expression were defined by 1.5-fold difference and a cutoff of P < 0.05 using limma-voom package. All processed data were statistically analyzed and evaluated by PANTHER classification system to determine the biological process function of genes in these lists. Compared to control, supplementation of recommended-dose AGP down-regulated genes related to inflammatory responses on d 5 and 11 PI; whereas, feeding low-dose AGP up-regulated genes associated with negative regulation of metabolic process on d 5, but down-regulated the genes related to immune responses on d 11 PI. The present observations support adverse effects of low-dose AGP in our previous study, indicated by exacerbated the detrimental effects of E. coli infection on pigs’ growth rate, diarrhea and systemic inflammation.

scholarly journals Measuring fast gene dynamics in single cells with time-lapse luminescence microscopy

2014 ◽  
Vol 25 (22) ◽  
pp. 3699-3708 ◽  
Author(s):  
Anyimilehidi Mazo-Vargas ◽  
Heungwon Park ◽  
Mert Aydin ◽  
Nicolas E. Buchler
Keyword(s):  
Gene Expression ◽  
Fluorescent Proteins ◽  
Single Cells ◽  
Time Lapse ◽  
Photon Flux ◽  
Luminescence Microscopy ◽  
Cell Cycle Genes ◽  
Light Excitation ◽  
Better Than

Time-lapse fluorescence microscopy is an important tool for measuring in vivo gene dynamics in single cells. However, fluorescent proteins are limited by slow chromophore maturation times and the cellular autofluorescence or phototoxicity that arises from light excitation. An alternative is luciferase, an enzyme that emits photons and is active upon folding. The photon flux per luciferase is significantly lower than that for fluorescent proteins. Thus time-lapse luminescence microscopy has been successfully used to track gene dynamics only in larger organisms and for slower processes, for which more total photons can be collected in one exposure. Here we tested green, yellow, and red beetle luciferases and optimized substrate conditions for in vivo luminescence. By combining time-lapse luminescence microscopy with a microfluidic device, we tracked the dynamics of cell cycle genes in single yeast with subminute exposure times over many generations. Our method was faster and in cells with much smaller volumes than previous work. Fluorescence of an optimized reporter (Venus) lagged luminescence by 15–20 min, which is consistent with its known rate of chromophore maturation in yeast. Our work demonstrates that luciferases are better than fluorescent proteins at faithfully tracking the underlying gene expression.

scholarly journals Gene expression polymorphism underpins evasion of host immunity in an asexual lineage of the Irish potato famine pathogen

2018 ◽  
Vol 18 (1) ◽  
Author(s):  
Marina Pais ◽  
Kentaro Yoshida ◽  
Artemis Giannakopoulou ◽  
Mathieu A. Pel ◽  
Liliana M. Cano ◽  
...  
Keyword(s):  
Gene Expression ◽  
Irish Potato ◽  
Host Immunity ◽  
Asexual Lineage ◽  
Potato Famine ◽  
Irish Potato Famine ◽  
Expression Polymorphism

scholarly journals Expression analysis of defense-related genes in cucumber (Cucumis sativus L.) against Phytophthora melonis

2020 ◽  
Author(s):  
Lida Hashemi ◽  
Ahmad Reza Golparvar ◽  
Mehdi Nasr Esfahani ◽  
Maryam Golabadi
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Molecular Mechanisms ◽  
Crown Rot ◽  
Economic Losses ◽  
Post Inoculation ◽  
Expression Ratio ◽  
Phytophthora Melonis ◽  
Maximum Expression

AbstractPhytophthora melonis is the causal agent of damping-off or crown rot, one of the most destructive cucumber diseases that causes severe economic losses in Iran and some other parts of the world. Despite intense research efforts made in the past years, no permanent cure currently exists for this disease. With the aim to understand the molecular mechanisms of defense against P. melonis, root collars and leaves of four cucumber genotypes consisting of resistant Ramezz; moderately resistant Baby and very susceptible Mini 6-23 and Extrem, were monitored for quantitative gene expression analysis of five antifungal and/or anti-oomycete genes (CsWRKY20, CsLecRK6.1, PR3, PR1-1a and LOX1) at three points after inoculation with P. melonis. The gene expression analysis indicated that P. melonis strongly enhanced the expression of these genes after inoculation in both leaves and root collars. Further, not only the transcript levels of these genes were significantly higher in the resistant and moderately resistance genotypes, but also the time point of the highest relative expression ratio for the five genes was different in the four cucumber genotypes. CsWRKY20 and PR3 showed the maximum expression in Ramezz at 48 hours post inoculation (hpi) while CsLecRK6.1, and LOX1 showed the highest expression at 72 hpi. In addition, PR1-1a showed the maximum expression in the Baby at 72 hpi. Root collars responded faster than leaves and some responses were more strongly up-regulated in root collars than in leaves. The genes found to be involved in disease resistance in two different organs of cucumber after pathogen infection. The results suggest that increased expression of these genes led to activation of defense pathways and could be responsible for a reduced P. melonis colonization capacity in Ramezz and Baby. Overall, this work represents a valuable resource for future functional genomics studies to unravel the molecular mechanisms of C. sativus- P. melonis interaction.

Differential Survival of Generic E. coli and Listeria spp. in Northeastern U.S. Soils Amended with Dairy Manure Compost, Poultry Litter Compost, and Heat-Treated Poultry Pellets and Fate in Raw Edible Radish Crops

2021 ◽  
Author(s):  
Marie Limoges ◽  
Deborah A. Neher ◽  
Thomas R. Weicht ◽  
Patricia D. Millner ◽  
Manan Sharma ◽  
...  
Keyword(s):  
Fresh Produce ◽  
Poultry Litter ◽  
Dairy Manure ◽  
Animal Origin ◽  
Most Probable Number ◽  
Post Inoculation ◽  
Probable Number ◽  
E Coli ◽  
Manure Compost ◽  
Heat Treated

Composted or heat-treated Biological Soil Amendments of Animal Origin (BSAAO) can be added to soils to provide nutrients for fresh produce. These products lower the risk of pathogen contamination of fresh produce when compared with use of untreated BSAAO; however, meteorological conditions, geographic location, and soil properties can influence the presence of pathogenic bacteria, or their indicators (e.g., generic E. coli) and allow potential for produce contamination. Replicated field plots of loamy or sandy soils were tilled and amended with dairy manure compost (DMC), poultry litter compost (PLC), or no compost (NoC) over two different field seasons, and non-composted heat-treated poultry pellets (HTPP) during the second field season. Plots were inoculated with a three-strain cocktail of rifampicin-resistant E. coli (rE.coli) at levels of 8.7 log CFU/m2. Direct plating and most probable number (MPN) methods measured the persistence of rE.coli and Listeria spp. in plots through 104 days post-inoculation. Greater survival of rE. coli was observed in PLC plots in comparison to DMC plots and NoC plots during year 1 (P < 0.05). Similar trends were observed for year 2, where rE. coli survival was also greater in HTPP amended plots (P < 0.05). Survival of rE. coli was dependent on soil type, where water potential and temperature were significant covariables. Listeria spp. were found in NoC plots, but not in plots amended with HTPP, PLC or DMC. Radish data demonstrate that PLC treatment promoted the greatest level of rE.coli translocation when compared to DMC and NoC treatments (P  < 0.05). These results are consistent with findings from studies conducted in other regions of the US and informs Northeast produce growers that composted and non-composted poultry-based BSAAO supports greater survival of rE. coli in field soils. This result has the potential to impact the food safety risk of edible produce grown in BSAAO amended soils as a result of pathogen contamination.

scholarly journals Vitamin B6 Is Required for Full Motility and Virulence in Helicobacter pylori

mBio ◽  
2010 ◽  
Vol 1 (3) ◽  
Author(s):  
Alexandra Grubman ◽  
Alexandra Phillips ◽  
Marie Thibonnier ◽  
Maria Kaparakis-Liaskos ◽  
Chad Johnson ◽  
...  
Keyword(s):  
Gene Expression ◽  
Helicobacter Pylori ◽  
Virulence Factors ◽  
Bacterial Pathogens ◽  
Therapeutic Targets ◽  
Vitamin B ◽  
Human Pathogens ◽  
Chronic Colonization ◽  
H Pylori

ABSTRACTDespite recent advances in our understanding of howHelicobacter pyloricauses disease, the factors that allow this pathogen to persist in the stomach have not yet been fully characterized. To identify new virulence factors inH. pylori, we generated low-infectivity variants of a mouse-colonizingH. pyloristrain using the classical technique ofin vitroattenuation. The resulting variants and their highly infectious progenitor bacteria were then analyzed by global gene expression profiling. The gene expression levels of five open reading frames (ORFs) were significantly reduced in low-infectivity variants, with the most significant changes observed for ORFs HP1583 and HP1582. These ORFs were annotated as encoding homologs of theEscherichia colivitamin B6biosynthesis enzymes PdxA and PdxJ. Functional complementation studies withE. coliconfirmedH. pyloriPdxA and PdxJ to bebona fidehomologs of vitamin B6biosynthesis enzymes. Importantly,H. pyloriPdxA was required for optimal growthin vitroand was shown to be essential for chronic colonization in mice. In addition to having a well-known metabolic role, vitamin B6is necessary for the synthesis of glycosylated flagella and for flagellum-based motility inH. pylori. Thus, for the first time, we identify vitamin B6biosynthesis enzymes as novel virulence factors in bacteria. Interestingly,pdxAandpdxJorthologs are present in a number of human pathogens, but not in mammalian cells. We therefore propose that PdxA/J enzymes may represent ideal candidates for therapeutic targets against bacterial pathogens.IMPORTANCEApproximately half of the world’s population is infected withH. pylori, yet howH. pyloribacteria establish chronic infections in human hosts remains elusive. From gene array studies, we identified two genes as representing potentially novel colonization factors forH. pylori. These genes encoded enzymes involved in the synthesis of vitamin B6, an important molecule for many metabolic reactions in living organisms. Little is currently known regarding vitamin B6biosynthesis in human pathogens. We showed that mutantH. pyloribacteria lacking an enzyme involved inde novovitamin B6biosynthesis, PdxA, were unable to synthesize motility appendages (flagella) and were unable to establish chronic colonization in mice. Thus, this work identifies vitamin B6biosynthesis enzymes as novel virulence factors for bacterial pathogens. Interestingly, a number of human pathogens, but not their mammalian hosts, possess these genes, which suggests that Pdx enzymes may represent ideal candidates for new therapeutic targets.

scholarly journals Thermal Resistance and Gene Expression of both Desiccation-Adapted and Rehydrated Salmonella enterica Serovar Typhimurium Cells in Aged Broiler Litter

2017 ◽  
Vol 83 (12) ◽  
Author(s):  
Zhao Chen ◽  
Xiuping Jiang
Keyword(s):  
Gene Expression ◽  
Heat Treatment ◽  
Thermal Resistance ◽  
High Temperatures ◽  
Human Pathogens ◽  
Wild Type ◽  
Broiler Litter ◽  
Content Type ◽  
Serovar Typhimurium ◽  
Trehalose Synthesis

ABSTRACT The objective of this study was to investigate the thermal resistance and gene expression of both desiccation-adapted and rehydrated Salmonella enterica serovar Typhimurium cells in aged broiler litter. S. Typhimurium was desiccation adapted in aged broiler litter with a 20% moisture content (water activity [aw], 0.81) for 1, 2, 3, 12, or 24 h at room temperature and then rehydrated for 3 h. As analyzed by quantitative real-time reverse transcriptase PCR (qRT-PCR), the rpoS, proV, dnaK, and grpE genes were upregulated (P < 0.05) under desiccation stress and could be induced after 1 h but in less than 2 h. Following rehydration, fold changes in the levels of these four genes became significantly lower (P < 0.05). The desiccation-adapted ΔrpoS mutant was less heat resistant at 75°C than was the desiccation-adapted wild type (P < 0.05), whereas there were no differences in heat resistance between desiccation-adapted mutants in two nonregulated genes (otsA and PagfD) and the desiccation-adapted wild type (P > 0.05). Survival characteristics of the desiccation-adapted ΔPagfD (rdar [red, dry, and rough] morphotype) and ΔagfD (saw [smooth and white] morphotype) mutants were similar (P > 0.05). Trehalose synthesis in the desiccation-adapted wild type was not induced compared to a nonadapted control (P > 0.05). Our results demonstrated the importance of the rpoS, proV, dnaK, and grpE genes in the desiccation survival of S. Typhimurium. By using an ΔrpoS mutant, we found that the rpoS gene was involved in the cross-protection of desiccation-adapted S. Typhimurium against high temperatures, while trehalose synthesis or rdar morphology did not play a significant role in this phenomenon. In summary, S. Typhimurium could respond rapidly to low-aw conditions in aged broiler litter while developing cross-protection against high temperatures, but this process could be reversed upon rehydration. IMPORTANCE Physical heat treatment is effective in eliminating human pathogens from poultry litter used as biological soil amendments. However, prior to physical heat treatment, some populations of microorganisms may be adapted to the stressful conditions in poultry litter during composting or stockpiling, which may cross-protect them against subsequent high temperatures. Our previous study demonstrated that desiccation-adapted S. enterica cells in aged broiler litter exhibited enhanced thermal resistance. However, there is limited research on the underlying mechanisms of the extended survival of pathogens under desiccation conditions in animal wastes and cross-tolerance to subsequent heat treatment. Moreover, no information is available about the thermal resistance of desiccation-adapted microorganisms in response to rehydration. Therefore, in the present study, we investigated the gene expression and thermal resistance of both desiccation-adapted and rehydrated S. Typhimurium in aged broiler litter. This work will guide future research efforts to control human pathogens in animal wastes used as biological soil amendments.

Analysis of Native Microflora and Selection of Strains Antagonistic to Human Pathogens on Fresh Produce†

2001 ◽  
Vol 64 (8) ◽  
pp. 1110-1115 ◽  
Author(s):  
CHING-HSING LIAO ◽  
WILLIAM F. FETT
Keyword(s):  
Fresh Produce ◽  
Brain Heart Infusion ◽  
Erwinia Carotovora ◽  
Plate Count ◽  
Human Pathogens ◽  
Green Pepper ◽  
Significant Difference ◽  
Bell Peppers ◽  
Native Microflora ◽  
Baby Carrots

The native microflora of three types of produce (green bell peppers, Romaine lettuce, and prepeeled baby carrots) and two types of sprouting seeds (alfalfa and clover) were investigated. Aerobic plate count (APC) for each produce or seed type as determined on Pseudomonas agar F (PAF) with incubation at 28°C was in the range of 4 to 7 log CFU per g of tissue or seed. There was no significant difference (P ≥ 0.05) in APC when the determinations were made with three agar media including PAF, brain heart infusion agar, and plate count agar. However, the APC as determined from plates that were incubated at 28°C was significantly (P ≤ 0.05) higher than with incubation at 37°C. Fluorescent pseudomonads accounted for 23 to 73% of APC and 6 to 18% of APC recovered from carrots, pepper, and lettuce were pectolytic. Forty-eight strains of pectolytic bacteria were randomly isolated and identified, respectively, as members of the genera of Pseudomonas, Erwinia, Bacillus, Xanthomonas, or Flavobacterium. Lactic acid bacteria and/or yeast were consistently isolated from baby carrots, lettuce, and sprouting seeds (alfalfa or clover) but not from green bell peppers. Approximately 120 strains of indigenous microflora were tested for their ability to inhibit the growth of Salmonella Chester, Listeria monocytogenes, Escherichia coli, or Erwinia carotovora subsp. carotovora on PAF. Six isolates capable of inhibiting the growth of at least one pathogen were isolated and identified, respectively, as Bacillus spp. (three strains), Pseudomonas aeruginosa (one strain), Pseudomonas fluorescens (strain A3), and yeast (strain D1). When green pepper disks were inoculated with strains A3 and D1, the growth of Salmonella Chester and L. monocytogenes on the disks was reduced by 1 and 2 logs, respectively, over a period of 3 days. Application of strains A3 and D1 as potential biopreservatives for enhancing the quality and safety of fresh produce is discussed.

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