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1.
Res Vet Sci ; 164: 105027, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37776608

ABSTRACT

Rickettsia africae causes zoonotic African tick bite fever, which is a disease of "One Health" importance. There have been reported cases of tourists from Europe and Asia who have been bitten by ticks whilst visiting South Africa's nature reserves, and on their return to their countries, the display African Tick Bite Fever sickness. Hence, the aim of this study was to determine the occurrence of Rickettsia africae in Amblyomma hebraeum ticks infesting livestock in the North West Province. A total of 358 A. hebraeum ticks were collected from 60 ruminants (cattle, sheep and goats) in Mafikeng City of North West Province, South Africa. Ticks were identified morphologically and further confirmed by sequencing of their ITS2 gene. DNA was extracted from 60 pools of ticks which consisted of 5-6 adult ticks that were from the same ruminant host. Infections with Rickettsia spp. were found in 48%, 40%, and 32% of cattle, sheep, and goats, respectively, in amplification by PCR using the ompA gene. The ompA gene sequences showed that the Rickettsia spp. were identified as R. africae. Although the animals from whom the ticks were collected did not exhibit any clinical symptoms, it is well recognised that R. africae is a disease with significant zoonotic potential. Thus, it is important to use the "One Health" approach to formulate prevention and control measures for this pathogen for animal and human health as well as the tourism sector due to the ecotourism importance of the resultant disease.


Subject(s)
Goat Diseases , Rickettsia Infections , Rickettsia , Sheep Diseases , Spotted Fever Group Rickettsiosis , Ticks , Animals , Cattle , Humans , Sheep , Amblyomma , Rickettsia Infections/epidemiology , Rickettsia Infections/veterinary , Rickettsia Infections/microbiology , South Africa/epidemiology , Rickettsia/genetics , Goats , Spotted Fever Group Rickettsiosis/veterinary , Goat Diseases/epidemiology , Sheep Diseases/epidemiology
2.
Sensors (Basel) ; 23(9)2023 Apr 29.
Article in English | MEDLINE | ID: mdl-37177600

ABSTRACT

The present work developed an electrochemical genosensor for the detection of virulence outer membrane protein A (ompA, tDNA) gene of Cronobacter sakazakii (C. sakazakii) by exploiting the excellent glucose-oxidase-mimicking activity of copper Metal-organic frameworks (Cu-MOF) doped with gold nanoparticle (AuNPs). The signal nanotags of signal probes (sDNA) that biofunctionalized AuNPs@Cu-MOF (sDNA-AuNPs@Cu-MOF) were designed using an Au-S bond. The biosensor was prepared by immobilization capture probes (cDNA) onto an electrodeposited AuNPs-modified glassy carbon electrode (GCE). AuNPs@Cu-MOF was introduced onto the surface of the GCE via a hybridization reaction between cDNA and tDNA, as well as tDNA and sDNA. Due to the enhanced oxidase-mimicking activity of AuNPs@Cu-MOF to glucose, the biosensor gave a linear range of 1.0 × 10-15 to 1.0 × 10-9 mol L-1 to tDNA with a detection limit (LOD) of 0.42 fmol L-1 under optimized conditions using differential pulse voltammetry measurement (DPV). It can be applied in the direct detection of ompA gene segments in total DNA extracts from C. sakazakii with a broad linear range of 5.4-5.4 × 105 CFU mL-1 and a LOD of 0.35 CFU mL-1. The biosensor showed good selectivity, fabricating reproducibility and storage stability, and can be used for the detection of ompA gene segments in real samples with recovery between 87.5% and 107.3%.


Subject(s)
Biosensing Techniques , Cronobacter sakazakii , Metal Nanoparticles , Metal-Organic Frameworks , Metal-Organic Frameworks/chemistry , Gold/chemistry , Copper/chemistry , DNA, Complementary , Glucose Oxidase , Reproducibility of Results , Limit of Detection , Metal Nanoparticles/chemistry , Carbon/chemistry , Glucose , Electrochemical Techniques , Electrodes
3.
Food Chem ; 391: 133268, 2022 Oct 15.
Article in English | MEDLINE | ID: mdl-35640339

ABSTRACT

This work presented an electrochemical biosensor for the detection of virulence outer membrane protein A (ompA) gene of Cronobacter sakazakii (C. sakazakii), which was based on the mimic peroxidase activity of boron doped quantum dots-Au nanoparticles (BQDs-AuNPs) and a signal amplification strategy of exonuclease III (Exo III)-assisted target-recycling (EATR). The electrochemical signal was come from the electrochemical reduction of H2O2 by BQDs-AuNPs nanozyme. Due to the enhanced peroxidase-mimic electrocatalytic efficiency of BQDs-AuNPs and the EATR strategy, the biosensor showed a broad linear range (1.0 × 10-15 - 1.0 × 10-8 mol L-1) and a low limit of detection (LOD, 4.0 × 10-17 mol L-1). The constructed biosensor could also be applied in direct detection of extracted DNA from C. sakazakii. A good linear relationship (7.8 - 7.8 × 106 CFU mL-1) between the logarithm concentration of C. sakazakii and electrochemical signal was obtained with a LOD of 2.6 CFU mL-1. The biosensor was applied in the detection of impA gene segments in contaminated infant formula with recoveries ranged in 83.4 - 108.2%.


Subject(s)
Biosensing Techniques , Cronobacter sakazakii , Metal Nanoparticles , Quantum Dots , Bacterial Outer Membrane Proteins , Boron , Carbon , Electrochemical Techniques , Exodeoxyribonucleases , Gold , Humans , Hydrogen Peroxide , Infant , Peroxidases , Virulence
4.
Arch Microbiol ; 202(3): 473-482, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31705142

ABSTRACT

The aim of this study is to evaluate the antibiofilm and antibacterial effects of auranofin against WT-ETBF, rETBF, WT-NTBF and clinically isolated Bacteroides fragilis strains. The minimum inhibitory concentration and biofilm inhibitory concentration of 0.25 µg/ml for auranofin against B. fragilis were determined, and the biofilm eradication concentration was 1 µg/ml. At an auranofin concentration of 0.5 µg/ml, little cellular metabolic activity was found. Confocal laser scanning microcopy results confirmed the inhibition of biofilm by auranofin. The effects of auranofin on the outer membrane protein (ompA) gene and the RND-type efflux pump (bmeB3) gene were investigated using quantitative real-time polymerase chain reaction (qRT-PCR). The qRT-PCR results showed that treatment with auranofin significantly reduced the gene expression compared to controls without auranofin. These data indicate the applicability of auranofin as a repurposed drug due to its inhibitory effect against biofilm formation of B. fragilis. Therefore, our study demonstrates that auranofin, already approved for human use, is a promising drug that has strong antibiofilm and antibacterial activity against B. fragilis.


Subject(s)
Anti-Bacterial Agents/pharmacology , Auranofin/pharmacology , Bacteroides Infections/microbiology , Bacteroides fragilis/drug effects , Biofilms/drug effects , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Bacteroides Infections/drug therapy , Bacteroides fragilis/genetics , Bacteroides fragilis/physiology , Drug Repositioning , Gene Expression Regulation, Bacterial/drug effects , Humans , Microbial Sensitivity Tests
5.
Iran J Basic Med Sci ; 22(6): 669-675, 2019 Jun.
Article in English | MEDLINE | ID: mdl-31231495

ABSTRACT

OBJECTIVES: Acinetobacter baumannii is one the most dangerous opportunistic pathogens in hospitalized infections. This bacterium is resistant to 90% of commercial antibiotics. Therefore, developing new strategies to cure A. baumannii-infections is urgent. The DNA vaccines new approach in which the immunogen can be directly expressed inside the target cells through cloning of immunogen into an expression vector. The outer membrane protein A(OmpA) is one the critical factors in pathogenicity of A. baumannii which has been repeatedly described as a powerful immunogen to trigger the immune responses. As the pure form of the OmpA is insoluble, vaccine delivery is very hard. MATERIALS AND METHODS: We previously cloned the ompA gene from A. baumannii into the eukaryotic expression vector pBudCE4.1 and observed that the OmpA protein has been considerably expressed in eukaryotic cell model. In current study, the immunogenic potential of pBudCE4.1-ompA has been evaluated in mice model of experimental. The serum levels of IgM, IgG, IL-2, IL-4, IL-12 and INF-γ were measured by enzyme-linked immunosorbent assay (ELISA) after immunization with ompA-vaccine. The protective efficiency of the designed-DNA vaccine was evaluated following intranasal administration of mice with toxic dose of A. baumannii. RESULTS: Obtained data showed the elevated levels of IgM, IgG, IL-2, IL-4, IL-12 and INF-γ in serum following the vaccine administration and mice who immunized with recombinant vector were survived more than control group. CONCLUSION: These findings indicate ompA-DNA vaccine is potent to trigger humoral and cellular immunity responses although further experiments are needed.

6.
New Microbes New Infect ; 25: 3-6, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29988839

ABSTRACT

Rickettsia felis, the causative agent of flea-borne spotted fever, occurs on all continents except Antarctica, owing to the cosmopolitan distribution of its cat flea vector. In this study, cat fleas were collected in two countries where the occurrence of R. felis was either unknown (Malta) or where accurate prevalence data were lacking (Israel). Altogether 129 fleas were molecularly analysed for the presence of rickettsial DNA. On the basis of three genetic markers, R. felis was identified in 39.5% (15/38) of the cat fleas from Malta. Sequences showed 100% identity to each other and to relevant sequences in GenBank. Among the 91 cat fleas from Israel, two (2.2%) contained the DNA of Candidatus Rickettsia senegalensis. Phylogenetically, the R. felis and Candidatus R. senegalensis identified here clustered separately (with high support) but within one clade, which was a sister group to that formed by the typhus group and spotted fever group rickettsiae. This is the first record of R. felis in Malta and of Candidatus R. senegalensis outside its formerly reported geographical range including Africa, Asia and North America.

7.
Indian J Microbiol ; 58(2): 174-181, 2018 Jun.
Article in English | MEDLINE | ID: mdl-29651176

ABSTRACT

Antibiotic resistant features of Acinetobacter baumannii is partly due to the decreased outer membrane proteins (OMPs) permeability. The OmpA is one of the most conserved proteins among A. baumannii with a considerable antigenic potential to stimulate the multidimensional immune system responses. The present study was aimed to clone the ompA gene into the eukaryotic expression vector with potential as DNA vaccine. The ompA gene of A. baumannii was amplified using polymerase chain reaction (PCR). The target DNA was cloned and sub-cloned into the pTZ57R/T and pBudCE4.1 vectors, respectively. The recombinant vectors containing ompA were then validated using colony PCR, vector sequencing and double-digestion strategies. The pBudCE4.1-ompA recombinant plasmid was transfected into the human dermal fibroblast cells (HDF) and presence of ompA transcript and protein was evaluated using reverse transcribed-PCR (RT-PCR) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Our finding from colony PCR, sequencing and enzyme double digestion result confirmed that target gene has been successfully inserted into the pTZ57RT and pBudCE4.1. The presence of an expected band (1112 bp) in RT-PCR as wells as a ~ 38 kDa band during SDS-PAGE showed that the recombinant pBudCE4.1-ompA construct was efficiently transfected into the HDF cells and expressed. Altogether, our observation demonstrated that the recombinant pBudCE4.1-ompA construct was successfully produced although further experiments are needed.

8.
Infect Genet Evol ; 57: 1-7, 2018 01.
Article in English | MEDLINE | ID: mdl-29107656

ABSTRACT

Tick-borne rickettsioses is one of the oldest known vector-borne diseases and has been viewed as emerging or re-emerging disease in China. The causative agents have been increasingly recognized and exhibited a high degree of genetic diversity and widespread distribution. This study was conducted to determine the prevalence of spotted fever group rickettsiae in ticks from Qinghai Province, northwestern China. In total, 860 questing adult ticks representing six species were collected. The SFG rickettsiae were detected in Haemaphysalis qinghaiensis (19.6%, 79/404), Dermacentor abaensis (73.7%, 157/213), Dermacentor silvarum (50.0%, 47/94), Dermacentor nuttalli (67.4%, 97/144), and Ixodes crenulatus (100%, 3/3), with an overall infection rate of 44.5%. The infection rates of SFG rickettsiae were significantly higher in Dermacentor spp. than in Haemaphysalis spp. (p<0.05). Sequence analyses of the gltA and ompA genes revealed that five SFG rickettsiae are present in ticks in Qinghai, including R. sibirica subspecies sibirica, R. raoultii, "Candidatus Rickettsia tibetani", and "Candidatus Rickettsia gannanii" Y27 and F107. Moreover, a potential novel Rickettsia species (Rickettsia sp. 10CYF) was identified in D. nuttalli and I. crenulatus. These findings extend our knowledge of the potential vector spectrum and distribution of rickettsiae, and provided valuable information for assessing the potential risk for public health.


Subject(s)
Rickettsia/classification , Rickettsia/genetics , Spotted Fever Group Rickettsiosis/epidemiology , Spotted Fever Group Rickettsiosis/microbiology , Ticks/microbiology , Animals , China/epidemiology , Genes, Bacterial , Humans , Phylogeny , Prevalence
9.
APMIS ; 124(12): 1109-1115, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27859690

ABSTRACT

To determine the prevalence and distribution of Chlamydia trachomatis genovars in patients with infertility by PCR-RFLP and ompA gene sequencing. Prevalence of other etiological agents (viz., Ureaplasma spp. and Mycoplasma hominis) were also assessed. Endocervical swabs were collected from 477 women and urine was collected from 151 men attending the Infertility Clinic. The samples were screened for C. trachomatis by cryptic plasmid, ompA gene and nested ompA gene PCR. Genotyping was performed by PCR-RFLP and sequencing. Samples were screened for Ureaplasma spp. and M. hominis. The prevalence of C. trachomatis in infertile women and their male partners were 15.7% (75 of 477) and 10.0% (15 of 151) respectively. Secondary infertility was significantly associated with chlamydial infection. Genovar E was the most prevalent followed by genovar D and F. Twenty-four C. trachomatis strains were selected for ompA gene sequencing. No mixed infection was picked. Variability in ompA sequences was seen in 50.0%. Both PCR-RFLP and ompA gene sequencing showed concordant results. High prevalence of C. trachomatis in infertile couples warrants routine screening for C. trachomatis infection in all infertile couples. Genotyping of the ompA gene of C. trachomatis may be a valuable tool in understanding the natural history of C. trachomatis infection.


Subject(s)
Chlamydia Infections/epidemiology , Chlamydia trachomatis/classification , Chlamydia trachomatis/isolation & purification , Genotype , Infertility/etiology , Adult , Bacterial Outer Membrane Proteins/genetics , Chlamydia trachomatis/genetics , Female , Genotyping Techniques , Humans , India/epidemiology , Male , Mycoplasma Infections/epidemiology , Mycoplasma hominis/isolation & purification , Pilot Projects , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Prevalence , Sequence Analysis, DNA , Ureaplasma/isolation & purification , Ureaplasma Infections/epidemiology , Young Adult
10.
Clin Microbiol Infect ; 22(6): 574.e7-9, 2016 Jun.
Article in English | MEDLINE | ID: mdl-27040805

ABSTRACT

The French Reference Centre for chlamydiae uses two real-time PCRs targeting the pmpH gene of Chlamydia trachomatis to differentiate between L strains and variant L2b, responsible for a lymphogranuloma venereum outbreak in Europe. We compared the results obtained for 122 L2b C. trachomatis-positive specimens, using the two real-time PCRs, with the sequencing of the ompA gene. Only 91 specimens were confirmed as L2b. Our results demonstrate that the lymphogranuloma venereum outbreak is no longer dominated by the variant L2b, and that many L-positive specimens were misidentified as L2b with the method used, which raises the question of its specificity.


Subject(s)
Bacterial Outer Membrane Proteins/genetics , Chlamydia trachomatis/isolation & purification , Diagnostic Errors , Lymphogranuloma Venereum/diagnosis , Molecular Diagnostic Techniques/methods , Real-Time Polymerase Chain Reaction/methods , Disease Outbreaks , Europe/epidemiology , Humans , Lymphogranuloma Venereum/epidemiology , Male , Sensitivity and Specificity , Sequence Analysis, DNA
11.
Parasit Vectors ; 9: 146, 2016 Mar 12.
Article in English | MEDLINE | ID: mdl-26968160

ABSTRACT

BACKGROUND: Rickettsia spp. are obligate intracellular bacteria and well known as transmitted by arthropods. These pathogens have a broad geographic distribution and a high degree of biological and clinical diversity. This study was conducted to determine the prevalence and molecular characterization of Rickettsia spp. in ticks collected from Gansu, where Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum were previously reported in ticks and ruminants. METHODS: A total of 1,583 questing Haemaphysalis qinghaiensis ticks were collected and tested for the presence of Rickettsia spp. gltA gene by PCR. Samples positive for gltA were examined by specific primers targeted for the ompA gene of SFG rickettsiae. The infections were further validated by sequencing and positive samples were genetically characterized based on the gltA and ompA genes. RESULTS: In total, Rickettsia spp. infection was found in 179 (18.5 %) H. qinghaiensis tick pools by using PCR and primers specific for the gltA gene. Of those, 157 (16.3 %) tick pools were positive for SFG rickettsiae by PCR based on ompA gene. Amplification and molecular analysis of the nucleotide sequences of gltA and ompA genes indicated three potential novel spotted fever group rickettsiae in H. qinghaiensis ticks. These three potential novel spotted fever group rickettsiae were clustered together in a subgroup, which represents a sister taxon to and separates from other known four SFG rickettsiae subgroups. CONCLUSIONS: This study revealed a high infection rate of SFG rickettsiae in H. qinghaiensis ticks in northwest China. Three potential novel spotted fever group rickettsiae classified into a novel SFG rickettsiae subgroup were identified and named "Candidatus Rickettsia gannanii" related strains in recognition of the location where it was first detected.


Subject(s)
Ixodidae/microbiology , Rickettsia/classification , Rickettsia/isolation & purification , Animals , China , Cluster Analysis , DNA, Bacterial , Genes, Bacterial , Phylogeny , Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology
12.
Iran J Vet Res ; 17(4): 281-285, 2016.
Article in English | MEDLINE | ID: mdl-28224015

ABSTRACT

Avian chlamydiosis is caused by Chlamydiophila psittaci with the highest infection rate in parrots (Psittacidae) and pigeons (Columbiformes). A two-year-old Congo African grey parrot was examined since the bird had shown clinical signs of anorexia, depression, diarrhea, and mild dyspnea and based on biochemical and hemathological analysis the bird was diagnosed as having anemia, leukocytosis, heterophilia, lymphopenia and monocytosis. With regards to clinical and paraclinical findings, the case was diagnosed to be carrying Chlamydiophila spp. In addition, choanal cleft and cloaca swabs were positive for Chlamydiophila spp. in a diagnostic polymerase chain reaction (PCR) (600 bp amplicon). Polymerase chain reaction products were typed by ompA gene-based PCR, using CTU/CTL primers (1050 bp amplicon). The PCR product sequence was compared with the sequences obtained from GenBank. The phylogenetic tree has revealed 100% identity with genotype B obtained from previous studies. The bird was hospitalized and treated with doxycycline regimen for 45 days, with a weekly sampling process to trace the presence of C. psittaci DNA in faecal and choanal swabs, this process continued to the point where the specimens turned negative after two weeks. Laboratory and radiology results were within normal limits after the treatment. Genotype B is predominantly isolated from Columbidae and there have not been any reports regarding the clinically affected African gray parrot with this genotype. Subsequently, to the best of our knowledge, this is the first report of chlamydiosis by genotype B on Congo African grey parrot.

13.
Infection ; 43(6): 739-41, 2015 Dec.
Article in English | MEDLINE | ID: mdl-25940434

ABSTRACT

Chlamydia trachomatis genogroups using ompA and multilocus sequence typing (MLST) were determined in consecutive isolates from school students aged 18 or older in the district of Brescia, Italy, 2012-2013. Among 40 samples, 4 ompA genovars and 18 STs were identified. Genovar E predominated (70 %) including five STs derived from ST59 (29 % of all isolates). This study, combining ompA and MLST typing of C. trachomatis school teenagers, suggests limited mixing and sexual interchange in this population.


Subject(s)
Chlamydia Infections/microbiology , Chlamydia trachomatis/classification , Chlamydia trachomatis/genetics , Genotype , Adolescent , Bacterial Outer Membrane Proteins/genetics , Chlamydia Infections/epidemiology , Chlamydia trachomatis/isolation & purification , Cross-Sectional Studies , Female , Humans , Italy/epidemiology , Male , Molecular Epidemiology , Multilocus Sequence Typing , Schools , Students
14.
Asian Pac J Trop Med ; 8(2): 94-7, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25902021

ABSTRACT

OBJECTIVE: To detect and characterize Chlamydophila psittaci (C. psittaci) in asymptomatic feral pigeons in central Thailand. METHODS: A total 814 swabs from the trachea and cloacae of 407 non-clinical feral pigeons in central Thailand were collected and tested for the presence of C. psittaci. RESULTS: A 10.8% of feral pigeons in the sample group were positive as determined by nested PCR primer specific to C. psittaci. The outer membrane protein A (ompA) gene of positive samples exhibited amino acid identity of C. psittaci ranging from 71 to 100% and were grouped in genotype B. Exceptionally, BF1676-56 isolate was closely related to Chlamydia avium with 99% identification of the 16S ribosomal (r) RNA gene. CONCLUSIONS: This is the first report on C. psittaci isolated from asymptomatic feral pigeons in Thailand, which provides knowledge for the disease status in pigeon populations in Thailand.

15.
Enferm Infecc Microbiol Clin ; 33(7): 458-63, 2015.
Article in Spanish | MEDLINE | ID: mdl-25533744

ABSTRACT

INTRODUCTION: Diagnosis of Chlamydia trachomatis infection in newborns is difficult; however, this diagnosis is performed by cell culture or by detection of IgM antibodies against C. trachomatis. Detection of C. trachomatis DNA in peripheral blood leukocytes using polymer chain reaction (PCR) may be a better tool for the diagnosis of infection by this pathogen. MATERIAL AND METHODS: A total of 44 premature newborns, all weighing less than 2500g, were included in the study. A blood sample and nasopharyngeal lavages were obtained from each newborn. Leukocyte DNA was obtained by phenol-chloroform extraction technique. Detection of C. trachomatis was performed by amplifying the ompA gene using the PCR endpoint. Cell culture tests and the detection of IgM antibodies against C. trachomatis by microimmunofluorescence assay were also performed. RESULTS: Twenty newborns were PCR-positive (45.5%), with this test being significantly associated with the presence of pneumonia (RR=2.28; 95%CI: 1.01 to 5.17; P=.035). The cell culture of nasopharyngeal lavage was positive in only 7 samples and no significant association was observed with any clinical or laboratory data. The titer of IgM antibodies against C. trachomatis associated with PCR-positive was 1:32 (RR=2.74; 95%CI: 1.21 to 6.23; P=.008), however this titer was not associated with the presence of pneumonia. CONCLUSION: DNA detection in peripheral blood leukocytes could be useful for diagnosis of C. trachomatis infection.


Subject(s)
Bacteremia/blood , Chlamydia trachomatis/isolation & purification , Chlamydial Pneumonia/blood , DNA, Bacterial/blood , Infant, Newborn/blood , Infant, Premature, Diseases/blood , Infant, Premature/blood , Leukocytes/microbiology , Antibodies, Bacterial/blood , Bacteremia/diagnosis , Bacteremia/microbiology , Bacterial Outer Membrane Proteins/genetics , Birth Weight , Chlamydia trachomatis/genetics , Chlamydia trachomatis/immunology , Chlamydial Pneumonia/diagnosis , Chlamydial Pneumonia/microbiology , Coinfection , Female , Humans , Immunoglobulin M/blood , Infant, Premature, Diseases/diagnosis , Infant, Premature, Diseases/microbiology , Male , Mycoplasma Infections/blood , Mycoplasma Infections/epidemiology , Nasopharynx/microbiology , Polymerase Chain Reaction/methods , Therapeutic Irrigation
16.
Article in English | WPRIM (Western Pacific) | ID: wpr-820395

ABSTRACT

OBJECTIVE@#To detect and characterize Chlamydophila psittaci (C. psittaci) in asymptomatic feral pigeons in central Thailand.@*METHODS@#A total 814 swabs from the trachea and cloacae of 407 non-clinical feral pigeons in central Thailand were collected and tested for the presence of C. psittaci.@*RESULTS@#A 10.8% of feral pigeons in the sample group were positive as determined by nested PCR primer specific to C. psittaci. The outer membrane protein A (ompA) gene of positive samples exhibited amino acid identity of C. psittaci ranging from 71 to 100% and were grouped in genotype B. Exceptionally, BF1676-56 isolate was closely related to Chlamydia avium with 99% identification of the 16S ribosomal (r) RNA gene.@*CONCLUSIONS@#This is the first report on C. psittaci isolated from asymptomatic feral pigeons in Thailand, which provides knowledge for the disease status in pigeon populations in Thailand.

17.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-500562

ABSTRACT

Objective:To detect and characterizeChlamydophila psittaci(C. psittaci) in asymptomatic feral pigeons in centralThailand.Methods:A total814 swabs from the trachea and cloacae of407 non-clinical feral pigeons in centralThailand were collected and tested for the presence ofC. psittaci.Results:A10.8% of feral pigeons in the sample group were positive as determined by nestedPCR primer specific toC. psittaci.The outer membrane proteinA(ompA) gene of positive samples exhibited amino acid identity ofC. psittaci ranging from71 to100% and were grouped in genotypeB.Exceptionally,BF1676-56 isolate was closely related toChlamydia avium with 99% identification of the16S ribosomal(r)RNA gene.Conclusions:This is the first report onC. psittaci isolated from asymptomatic feral pigeons inThailand, which provides knowledge for the disease status in pigeon populations inThailand.

18.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-951549

ABSTRACT

Objective: To detect and characterize Chlamydophila psittaci (C. psittaci) in asymptomatic feral pigeons in central Thailand. Methods: A total 814 swabs from the trachea and cloacae of 407 non-clinical feral pigeons in central Thailand were collected and tested for the presence of C. psittaci. Results: A 10.8% of feral pigeons in the sample group were positive as determined by nested PCR primer specific to C. psittaci. The outer membrane protein A (ompA) gene of positive samples exhibited amino acid identity of C. psittaci ranging from 71 to 100% and were grouped in genotype B. Exceptionally, BF1676-56 isolate was closely related to Chlamydia avium with 99% identification of the 16S ribosomal (r) RNA gene. Conclusions: This is the first report on C. psittaci isolated from asymptomatic feral pigeons in Thailand, which provides knowledge for the disease status in pigeon populations in Thailand.

19.
Vaccine ; 31(48): 5754-9, 2013 Nov 19.
Article in English | MEDLINE | ID: mdl-24012571

ABSTRACT

In the present study, poly (lactic-co-glycolic) acid (PLGA) was used as a carrier for a divalent fusion DNA vaccine encoding the Aeromonas veronii outer membrane protein A (ompA) and Aeromonas hydrophila hemolysins (hly) protein. The recombinant pET-28a-ompA-hly was constructed by inserting the ompA gene and hly gene into a pET-28a expression vector. Loading of ompA-hly antigen module on PLGA microspheres were accomplished by water-in-oil-in-water (W/O/W) encapsulation. The molecular weight and specificity of pET-28a-ompA-hly were detected by dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting. The microspheres showed an average particle size of 100-150 µm and a loading efficiency (LE) of 68.8%. Mice received ompA-hly antigen-loaded PLGA microspheres by intraperitoneal or intragastric administration mounted strong and sustained IgG response, which was significantly higher (p<0.05) than those achieved by pET-28a-ompA-hly antigen alone. OmpA-hly antigen-loaded PLGA microsphere vaccine uniquely conferred a long lasting (30 days) sterile immunity against challenge infection. Results indicated that ompA-hly antigen-loaded PLGA microsphere vaccine is a qualified candidate vector system for sterile protective immunity against A. hydrophila and A. veronii infections.


Subject(s)
Aeromonas/immunology , Bacterial Outer Membrane Proteins/immunology , Bacterial Vaccines/immunology , Drug Carriers/administration & dosage , Hemolysin Proteins/immunology , Lactic Acid/administration & dosage , Polyglycolic Acid/administration & dosage , Vaccines, DNA/immunology , Aeromonas/genetics , Animals , Antibodies, Bacterial/blood , Bacterial Outer Membrane Proteins/genetics , Bacterial Vaccines/administration & dosage , Bacterial Vaccines/genetics , Female , Hemolysin Proteins/genetics , Immunoglobulin G/blood , Mice , Mice, Inbred BALB C , Microspheres , Polylactic Acid-Polyglycolic Acid Copolymer , Time Factors , Vaccination/methods , Vaccines, DNA/administration & dosage , Vaccines, DNA/genetics , Vaccines, Synthetic/administration & dosage , Vaccines, Synthetic/genetics , Vaccines, Synthetic/immunology
20.
Rev. argent. microbiol ; 44(2): 65-68, jun. 2012. ilus, graf, tab
Article in Spanish | LILACS | ID: lil-657613

ABSTRACT

Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, Including humans, other mammals and birds. However, very little is known about chlamydial infections in birds in our region. In the present study, we examined 28 clinically normal birds In illegal captivity that were confiscated in the province of Córdoba, Argentina. The objective was to detect Chlamydophila spp. in cloacal swabs by genetic analysis of the ompA gene. Nested-PCR of the ompA gene identified five samples as Chlamydophila pecorum and the sequence analysis demonstrated the presence of the ompA gene of C. pecorum In these birds. On the other hand, Chlamydophila psittaci was not detected. These birds could be either asymptomatic reservoirs or subclinical carriers of C. pecorum. This is the first report of the detection of C. pecorum in Argentina.


Las bacterias que pertenecen a la familia Chlamydiaceae causan un extenso espectro de enfermedades en una amplia gama de huéspedes, incluidos los seres humanos, otros mamíferos y aves. Sin embargo, se sabe muy poco acerca de las infecciones por clamidias en aves de nuestra reglón. Esta Investigación examinó 28 aves clínicamente normales mantenidas en cautiverio ¡legal, que fueron confiscadas en Córdoba, Argentina. El objetivo fue detectar Chlamydophila spp. en hisopados cloacales por análisis del gen ompA. La PCR anidada del gen ompA reveló la presencia de Chlamydophila pecorum en cinco muestras. El análisis de secuencias demostró la presencia del gen ompA de C. pecorum en estas aves. Por el contrario, Chlamydophila psittaci no se detectó. Estas aves pueden ser reservónos asintomáticos o portadores subclínlcos de C. pecorum. Este es el primer informe de la detección de C. pecorum en la Argentina.


Subject(s)
Animals , Bacterial Outer Membrane Proteins/genetics , Carrier State/veterinary , Chlamydophila Infections/veterinary , Chlamydophila/genetics , Genes, Bacterial , Passeriformes/genetics , Amino Acid Sequence , Argentina/epidemiology , Carrier State/epidemiology , Carrier State/microbiology , Chlamydophila Infections/epidemiology , Chlamydophila Infections/microbiology , Chlamydophila/classification , Cloaca/microbiology , Molecular Sequence Data , Phylogeny , Sequence Alignment , Sequence Homology, Amino Acid , Species Specificity
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