Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 6 de 6
Filter
Add more filters










Database
Language
Publication year range
1.
PLoS Negl Trop Dis ; 17(11): e0011777, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37983257

ABSTRACT

BACKGROUND: There are limited longitudinal data on the acquisition of Giardia lamblia infections in childhood using molecular assays to detect and type assemblages, and measure effects of infections on diarrhea risk and childhood growth. METHODS: We analysed stool samples from a surveillance sample within a birth cohort in a rural district in tropical Ecuador. The cohort was followed to 8 years of age for the presence of G. lamblia in stools by quantitative PCR and A and B assemblages by Taqman assay or Sanger sequencing. We explored risk factors associated with infection using generalized estimating equations applied to longitudinal binary outcomes, and longitudinal panel data analysis to estimate effects of infection on diarrhea and growth trajectories. RESULTS: 2,812 stool samples collected between 1 month and 8 years of age from 498 children were analyzed and showed high rates of infection: 79.7% were infected at least once with peak prevalence (53.9%) at 5 years. Assemblage B was accounted for 56.8% of genotyped infections. Risk factors for infection included male sex (P = 0.001), daycare attendance (P<0.001), having a household latrine (P = 0.04), childhood (P<0.001) and maternal soil-transmitted helminth (P = 0.029) infections, and exposures to donkeys (age interaction P = 0.034). G. lamblia was associated with increased risk of diarrhea (per episode, RR 1.03, 95% CI 1.01-1.06, P = 0.011) during the first 3 years of life and a transient impairment of weight (age interaction P = 0.017) and height-for-age (age interaction P = 0.025) trajectories between 1 and 4 years of age. There was no increased risk of either assemblage being associated with outcomes. CONCLUSION: Our data show a relatively high edemicity of G. lamblia transmission during childhood in coastal Ecuador, and evidence that infection is associated with a transiently increased risk of diarrhea during the first 3 years of life and impairment of weight and height between 1 and 4 years.


Subject(s)
Giardia lamblia , Giardiasis , Child , Humans , Male , Child, Preschool , Infant, Newborn , Giardiasis/epidemiology , Giardia lamblia/genetics , Birth Cohort , Ecuador/epidemiology , Giardia/genetics , Diarrhea/epidemiology , Feces
2.
Parasit Vectors ; 13(1): 141, 2020 Mar 18.
Article in English | MEDLINE | ID: mdl-32188497

ABSTRACT

BACKGROUND: Enteric parasites are transmitted in households but few studies have sampled inside households for parasites and none have used sensitive molecular methods. METHODS: We collected bed and living room dust samples from households of children participating in a clinical trial of anthelmintic treatment in rural coastal Ecuador. Dust was examined for presence of DNA specific for 11 enteric parasites (Ascaris lumbricoides, Trichuris trichiura, Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Toxocara canis and T. cati, Giardia lamblia, Blastocystis hominis, Cryptosporidium spp., and Entamoeba histolytica) by quantitative PCR (qPCR). RESULTS: Of the 38 households sampled, 37 had positive dust for at least one parasite and up to 8 parasites were detected in single samples. Positivity was greatest for B. hominis (79% of household samples) indicating a high level of environmental fecal contamination. Dust positivity rates for individual pathogens were: S. stercoralis (52%), A. lumbricoides (39%), G. lamblia (39%), Toxocara spp. (42%), hookworm (18%) and T. trichiura (8%). DNA for Cryptosporidium spp. and E. histolytica was not detected. Bed dust was more frequently positive than floor samples for all parasites detected. Positivity for A. lumbricoides DNA in bed (adjusted OR: 10.0, 95% CI: 2.0-50.1) but not floor dust (adjusted OR: 3.6, 95% CI: 0.3-37.9) was significantly associated with active infections in children. CONCLUSIONS: To our knowledge, this is the first use of qPCR on environmental samples to detect a wide range of enteric pathogen DNA. Our results indicate widespread contamination of households with parasite DNA and raise the possibility that beds, under conditions of overcrowding in a humid tropical setting, may be a source of transmission.


Subject(s)
Beds/parasitology , DNA/analysis , Dust/analysis , Family Characteristics , Intestinal Diseases, Parasitic/transmission , Parasites/genetics , Adolescent , Animals , Child , Controlled Clinical Trials as Topic , Environment , Feces/parasitology , Female , Humans , Male , Real-Time Polymerase Chain Reaction , Rural Population
3.
Microb Biotechnol ; 11(2): 302-316, 2018 03.
Article in English | MEDLINE | ID: mdl-29027367

ABSTRACT

Bacillus pumilus strain 15.1 was previously found to cause larval mortality in the Med-fly Ceratitis capitata and was shown to produce crystals in association with the spore. As parasporal crystals are well-known as invertebrate-active toxins in entomopathogenic bacteria such as Bacillus thuringiensis (Cry and Cyt toxins) and Lysinibacillus sphaericus (Bin and Cry toxins), the B. pumilus crystals were characterized. The crystals were composed of a 45 kDa protein that was identified as an oxalate decarboxylase by peptide mass fingerprinting, N-terminal sequencing and by comparison with the genome sequence of strain 15.1. Synthesis of crystals by a plasmid-cured derivative of strain 15.1 (produced using a novel curing strategy), demonstrated that the oxalate decarboxylase was encoded chromosomally. Crystals spontaneously solubilized when kept at low temperatures, and the protein produced was resistant to trypsin treatment. The insoluble crystals produced by B. pumilus 15.1 did not show significant toxicity when bioassayed against C. capitata larvae, but once the OxdD protein was solubilized, an increase of toxicity was observed. We also demonstrate that the OxdD present in the crystals has oxalate decarboxylate activity as the formation of formate was detected, which suggests a possible mechanism for B. pumilus 15.1 activity. To our knowledge, the characterization of the B. pumilus crystals as oxalate decarboxylase is the first report of the natural production of parasporal inclusions of an enzyme.


Subject(s)
Bacillus pumilus/chemistry , Bacillus pumilus/pathogenicity , Bacterial Proteins/analysis , Carboxy-Lyases/analysis , Spores, Bacterial/chemistry , Spores, Bacterial/pathogenicity , Virulence Factors/analysis , Animals , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Biological Assay , Carboxy-Lyases/chemistry , Carboxy-Lyases/metabolism , Ceratitis capitata/drug effects , Ceratitis capitata/microbiology , Cold Temperature , Larva/drug effects , Mass Spectrometry , Proteolysis , Solubility , Survival Analysis , Virulence , Virulence Factors/chemistry , Virulence Factors/metabolism
4.
Appl Microbiol Biotechnol ; 100(8): 3637-54, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26782747

ABSTRACT

In the present work, the local isolate Bacillus pumilus 15.1 has been morphologically and biochemically characterized in order to gain a better understanding of this novel entomopathogenic strain active against Ceratitis capitata. This strain could represent an interesting biothechnological tool for the control of this pest. Here, we report on its nutrient preferences, extracellular enzyme production, motility mechanism, biofilm production, antibiotic suceptibility, natural resistance to chemical and physical insults, and morphology of the vegetative cells and spores. The pathogen was found to be ß-hemolytic and susceptible to penicillin, ampicillin, chloramphenicol, gentamicin, kanamycin, rifampicin, tetracycline, and streptomycin. We also report a series of biocide, thermal, and UV treatments that reduce the viability of B. pumilus 15.1 by several orders of magnitude. Heat and chemical treatments kill at least 99.9 % of vegetative cells, but spores were much more resistant. Bleach was the only chemical that was able to completely eliminate B. pumilus 15.1 spores. Compared to the B. subtilis 168 spores, B. pumilus 15.1 spores were between 2.67 and 350 times more resistant to UV radiation while the vegetative cells of B. pumilus 15.1 were almost up to 3 orders of magnitude more resistant than the model strain. We performed electron microscopy for morphological characterization, and we observed geometric structures resembling the parasporal crystal inclusions synthesized by Bacillus thuringiensis. Some of the results obtained here such as the parasporal inclusion bodies produced by B. pumilus 15.1 could potentially represent virulence factors of this novel and potentially interesting strain.


Subject(s)
Bacillus pumilus/physiology , Bacillus thuringiensis/metabolism , Ceratitis capitata/microbiology , Inclusion Bodies/metabolism , Animals , Bacillus pumilus/growth & development , Bacillus pumilus/radiation effects , Bacillus pumilus/ultrastructure , Bacillus thuringiensis/growth & development , Bacillus thuringiensis/radiation effects , Bacillus thuringiensis/ultrastructure , Inclusion Bodies/ultrastructure , Microscopy, Electron , Spores, Bacterial/metabolism , Spores, Bacterial/radiation effects , Spores, Bacterial/ultrastructure , Ultraviolet Rays
5.
Plasmid ; 82: 17-27, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26416357

ABSTRACT

The Bacillus pumilus 15.1 strain, a recently described entomopathogenic strain active against Ceratitis capitata, contains at least two extrachromosomal elements, pBp15.1S and pBp15.1B. Given that B. pumilus is not a typical entomopathogenic bacterium, the acquisition of this extrachromosomal DNA may explain why B. pumilus 15.1 is toxic to an insect. One of the plasmids present in the strain, the pBp15.1S plasmid, was sub-cloned, sequenced and analyzed using bioinformatics to identify any potential virulence factor. The pBp15.1S plasmid was found to be 7785 bp in size with a GC content of 35.7% and 11 putative ORFs. A replication module typical of a small rolling circle plasmid and a sensing and regulatory system specific for plasmids was found in pBp15.1S. Additionally, we demonstrated the existence of ssDNA in plasmid preparations suggesting that pBp15.1S replicates by the small rolling circle mechanism. A gene cluster present in plasmid pPZZ84 from a distantly isolated B. pumilus strain was also present in pBp15.1S. The plasmid copy number of pBp15.1S in exponentially growing B. pumilus cells was determined to be 33 copies per chromosome. After an extensive plasmid characterization, no known virulence factor was found so a search in the other extrachromosomal elements of the bacteria is needed.


Subject(s)
Bacillus/genetics , Gene Dosage/genetics , Open Reading Frames/genetics , Plasmids/genetics , Base Composition/genetics , Base Sequence , Cloning, Molecular , DNA, Single-Stranded/isolation & purification , Molecular Sequence Data , RNA, Untranslated/genetics , Sequence Analysis, DNA
6.
Genome Announc ; 3(5)2015 Sep 24.
Article in English | MEDLINE | ID: mdl-26404596

ABSTRACT

We present the draft whole-genome sequence of the entomopathogenic Bacillus pumilus 15.1 strain that consists of 3,795,691 bp and 3,776 predicted protein-coding genes. This genome sequence provides the basis for understanding the potential mechanism behind the toxicity and virulence of B. pumilus 15.1 against the Mediterranean fruit fly.

SELECTION OF CITATIONS
SEARCH DETAIL
...