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1.
Emerg Infect Dis ; 29(7): 1-9, 2023 07.
Article in English | MEDLINE | ID: mdl-37347519

ABSTRACT

We provide incidences (cases/10 million persons) in the Netherlands during 2009-2019 for pathogens listed as potential bioterrorism agents. We included pathogens from the highest categories of the European Medicines Agency or the US Centers for Disease Control and Prevention. Notifiable diseases and recently published data were used to calculate the average annual incidence. Coxiella burnetii had the highest incidence because of a Q fever epidemic during 2007-2010. Incidence then decreased to 10.8 cases/. Pathogens with an incidence >1 were Brucella spp. (2.5 cases), Francisella tularensis (1.3 cases), and Burkholderia pseudomallei (1.1 cases). Pathogens with an incidence <1 were hemorrhagic fever viruses (0.3 cases), Clostridium botulinum (0.2 cases), and Bacillus anthracis (0.1 cases). Variola major and Yersinia pestis were absent. The generally low incidences make it unlikely that ill-meaning persons can isolate these pathogens from natural sources in the Netherlands. However, the pathogens are stored in laboratories, underscoring the need for biosecurity measures.


Subject(s)
Bacillus anthracis , Francisella tularensis , Biological Warfare Agents , Bioterrorism/prevention & control , Netherlands/epidemiology
2.
Microorganisms ; 11(4)2023 Mar 30.
Article in English | MEDLINE | ID: mdl-37110328

ABSTRACT

Francisella tularensis is a zoonotic bacterium that is endemic in large parts of the world. It is absent in the standard library of the most applied matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) systems: the Vitek MS and the Bruker Biotyper system. The additional Bruker MALDI Biotyper Security library contains F. tularensis without subspecies differentiation. The virulence of F. tularensis differs between the subspecies. The F. tularensis subspecies (ssp.) tularensis is highly pathogenic, whereas the subspecies holarctica displays lower virulence and subspecies novicida and F. tularensis ssp. mediasiatica are hardly virulent. To differentiate the Francisellaceae and the F. tularensis-subspecies, an in-house Francisella library was built with the Bruker Biotyper system and validated together with the existing Bruker databases. In addition, specific biomarkers were defined based on the main spectra of the Francisella strains supplemented with in silico genome data. Our in-house Francisella library accurately differentiates the F. tularensis subspecies and the other Francisellaceae. The biomarkers correctly differentiate the various species within the genus Francisella and the F. tularensis subspecies. These MALDI-TOF MS strategies can successfully be applied in a clinical laboratory setting as a fast and specific method to identify F. tularensis to subspecies level.

3.
Clin Infect Dis ; 75(12): 2250-2252, 2022 12 19.
Article in English | MEDLINE | ID: mdl-35653425

ABSTRACT

A patient was diagnosed with Brucella canis following exposure to infected dogs in her breeding facility. Transboundary spread of B. canis through (illegal) import of infected dogs to non-endemic countries in Europe suggest that B. canis infection should be considered in European patients with occupational exposure to dogs.


Subject(s)
Brucella canis , Brucellosis , Dog Diseases , Animals , Dogs , Female , Humans , Brucellosis/diagnosis , Brucellosis/veterinary , Dog Diseases/diagnosis , Europe , Netherlands
4.
Emerg Infect Dis ; 28(4): 883-885, 2022 04.
Article in English | MEDLINE | ID: mdl-35318921

ABSTRACT

We used national registry data on human cases of Francisella tularensis subspecies holarctica infection to assess transmission modes among all 26 autochthonous cases in the Netherlands since 2011. The results indicate predominance of terrestrial over aquatic animal transmission sources. We recommend targeting disease-risk communication toward hunters, recreationists, and outdoor professionals.


Subject(s)
Francisella tularensis , Tularemia , Animals , Humans , Netherlands/epidemiology , Tularemia/epidemiology
5.
Microorganisms ; 10(2)2022 Feb 14.
Article in English | MEDLINE | ID: mdl-35208889

ABSTRACT

Shigella spp. and E. coli are closely related and cannot be distinguished using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) with commercially available databases. Here, three alternative approaches using MALDI-TOF MS to identify and distinguish Shigella spp., E. coli, and its pathotype EIEC were explored and evaluated using spectra of 456 Shigella spp., 42 E. coli, and 61 EIEC isolates. Identification with a custom-made database resulted in >94% Shigella identified at the genus level and >91% S. sonnei and S. flexneri at the species level, but the distinction of S. dysenteriae, S. boydii, and E. coli was poor. With biomarker assignment, 98% S. sonnei isolates were correctly identified, although specificity was low. Discriminating markers for S. dysenteriae, S. boydii, and E. coli were not assigned at all. Classification models using machine learning correctly identified Shigella in 96% of isolates, but most E. coli isolates were also assigned to Shigella. None of the proposed alternative approaches were suitable for clinical diagnostics for identifying Shigella spp., E. coli, and EIEC, reflecting their relatedness and taxonomical classification. We suggest the use of MALDI-TOF MS for the identification of the Shigella spp./E. coli complex, but other tests should be used for distinction.

6.
Travel Med Infect Dis ; 42: 102080, 2021.
Article in English | MEDLINE | ID: mdl-33933687

ABSTRACT

A 27 years-old Dutch male returning from Nepal presented with a painful abscess on the left forearm without fever or other systemic complications. Signs and symptoms consisted of culture of the abscess material revealed Burkholderia pseudomallei. Laboratory results, chest X-ray and CT scan of the abdomen were without abnormalities. The patient was initially treated with 2 weeks of ceftazidime and continued with a 6-week oral eradication phase with trimethoprim-sulfamethoxazole. The patient recovered without complications. Melioidosis is encountered relatively infrequently as an imported condition, mainly from Southeast Asia with focus on Thailand. Melioidosis from Nepal is a rarity and has previously been described in only four cases, with possible acquisition abroad in three of those.


Subject(s)
Burkholderia pseudomallei , Melioidosis , Adult , Anti-Bacterial Agents/therapeutic use , Ceftazidime/therapeutic use , Humans , Male , Melioidosis/diagnosis , Melioidosis/drug therapy , Nepal
7.
Front Microbiol ; 11: 564103, 2020.
Article in English | MEDLINE | ID: mdl-33193150

ABSTRACT

Shigella spp. and entero-invasive Escherichia coli (EIEC) can cause mild diarrhea to dysentery. In Netherlands, although shigellosis is a notifiable disease, there is no laboratory surveillance for Shigella spp. and EIEC in place. Consequently, the population structure for circulating Shigella spp. and EIEC isolates is not known. This study describes the phenotypic and serological characteristics, the phenotypic and genetic antimicrobial resistance (AMR) profiles, the virulence gene profiles, the classic multi-locus sequence types (MLST) and core genome (cg)MLST types, and the epidemiology of 414 Shigella spp. and EIEC isolates collected during a cross-sectional study in Netherlands in 2016 and 2017. S. sonnei (56%), S. flexneri (25%), and EIEC (15%) were detected predominantly in Netherlands, of which the EIEC isolates were most diverse according to their phenotypical profile, O-types, MLST types, and cgMLST clades. Virulence gene profiling showed that none of the isolates harbored Shiga toxin genes. Most S. flexneri and EIEC isolates possessed nearly all virulence genes examined, while these genes were only detected in approximately half of the S. sonnei isolates, probably due to loss of the large invasion plasmid upon subculturing. Phenotypical resistance correlated well with the resistant genotype, except for the genes involved in resistance to aminoglycosides. A substantial part of the characterized isolates was resistant to antimicrobials advised for treatment, i.e., 73% was phenotypically resistant to co-trimoxazole and 19% to ciprofloxacin. AMR was particularly observed in isolates from male patients who had sex with men (MSM) or from patients that had traveled to Asia. Furthermore, isolates related to international clusters were also circulating in Netherlands. Travel-related isolates formed clusters with isolates from patients without travel history, indicating their emergence into the Dutch population. In conclusion, laboratory surveillance using whole genome sequencing as high-resolution typing technique and for genetic characterization of isolates complements the current epidemiological surveillance, as the latter is not sufficient to detect all (inter)national clusters, emphasizing the importance of multifactorial public health approaches.

8.
BMC Genomics ; 21(1): 138, 2020 Feb 10.
Article in English | MEDLINE | ID: mdl-32041522

ABSTRACT

BACKGROUND: We investigated the association of symptoms and disease severity of shigellosis patients with genetic determinants of infecting Shigella and entero-invasive Escherichia coli (EIEC), because determinants that predict disease outcome per individual patient could be used to prioritize control measures. For this purpose, genome wide association studies (GWAS) were performed using presence or absence of single genes, combinations of genes, and k-mers. All genetic variants were derived from draft genome sequences of isolates from a multicenter cross-sectional study conducted in the Netherlands during 2016 and 2017. Clinical data of patients consisting of binary/dichotomous representation of symptoms and their calculated severity scores were also available from this study. To verify the suitability of the methods used, the genetic differences between the genera Shigella and Escherichia were used as control. RESULTS: The isolates obtained were representative of the population structure encountered in other Western European countries. No association was found between single genes or combinations of genes and separate symptoms or disease severity scores. Our benchmark characteristic, genus, resulted in eight associated genes and > 3,000,000 k-mers, indicating adequate performance of the algorithms used. CONCLUSIONS: To conclude, using several microbial GWAS methods, genetic variants in Shigella spp. and EIEC that can predict specific symptoms or a more severe course of disease were not identified, suggesting that disease severity of shigellosis is dependent on other factors than the genetic variation of the infecting bacteria. Specific genes or gene fragments of isolates from patients are unsuitable to predict outcomes and cannot be used for development, prioritization and optimization of guidelines for control measures of shigellosis or infections with EIEC.


Subject(s)
Dysentery, Bacillary/diagnosis , Dysentery, Bacillary/microbiology , Escherichia coli Infections/diagnosis , Escherichia coli Infections/microbiology , Escherichia coli/genetics , Shigella/genetics , Cross-Sectional Studies , Escherichia coli/classification , Escherichia coli/isolation & purification , Genetic Markers , Genome-Wide Association Study , Humans , Phylogeny , Shigella/classification , Shigella/isolation & purification
9.
BMC Infect Dis ; 19(1): 1037, 2019 Dec 09.
Article in English | MEDLINE | ID: mdl-31818261

ABSTRACT

BACKGROUND: Shigella spp. and entero-invasive E. coli (EIEC) use the same invasive mechanism to cause diarrheal diseases. Public health regulations apply only to Shigella spp. infections, but are hampered by the lack of simple methods to distinguish them from EIEC. In the last decades, molecular methods for detecting Shigella spp. and EIEC were implemented in medical microbiological laboratories (MMLs). However, shigellosis cases identified with molecular techniques alone are not notifiable in most countries. Our study investigates the impact of EIEC versus Shigella spp. infections and molecular diagnosed shigellosis versus culture confirmed shigellosis for re-examination of the rationale for the current public health regulations. METHODS: In this multicenter cross-sectional study, fecal samples of patients suspected for gastro-enteritis, referred to 15 MMLs in the Netherlands, were screened by PCR for Shigella spp. or EIEC. Samples were cultured to discriminate between the two pathogens. We compared risk factors, symptoms, severity of disease, secondary infections and socio-economic consequences for (i) culture-confirmed Shigella spp. versus culture-confirmed EIEC cases (ii) culture positive versus PCR positive only shigellosis cases. RESULTS: In 2016-2017, 777 PCR positive fecal samples with patient data were included, 254 of these were culture-confirmed shigellosis cases and 32 were culture-confirmed EIEC cases. EIEC cases were more likely to report ingestion of contaminated food and were less likely to be men who have sex with men (MSM). Both pathogens were shown to cause serious disease although differences in specific symptoms were observed. Culture-negative but PCR positive cases were more likely report travel or ingestion of contaminated food and were less likely to be MSM than culture-positive cases. Culture-negative cases were more likely to suffer from multiple symptoms. No differences in degree of secondary infections were observed between Shigella spp. and EIEC, and culture-negative and culture-positive cases. CONCLUSIONS: No convincing evidence was found to support the current guidelines that employs different measures based on species or detection method. Therefore, culture and molecular detection methods for Shigella spp. and EIEC should be considered equivalent for case definition and public health regulations regarding shigellosis. Differences were found regarding risks factors, indicating that different prevention strategies may be required.


Subject(s)
Dysentery, Bacillary/epidemiology , Dysentery, Bacillary/microbiology , Escherichia coli Infections/epidemiology , Escherichia coli Infections/microbiology , Adolescent , Adult , Bacteriological Techniques/methods , Cross-Sectional Studies , Diarrhea/microbiology , Dysentery, Bacillary/etiology , Escherichia coli/genetics , Escherichia coli/isolation & purification , Escherichia coli/pathogenicity , Escherichia coli Infections/etiology , Feces/microbiology , Female , Gastroenteritis/microbiology , Homosexuality, Male/statistics & numerical data , Humans , Incidence , Male , Middle Aged , Netherlands/epidemiology , Polymerase Chain Reaction , Public Health , Shigella/genetics , Shigella/isolation & purification , Shigella/pathogenicity , Young Adult
11.
J Clin Microbiol ; 56(10)2018 10.
Article in English | MEDLINE | ID: mdl-30021824

ABSTRACT

Identification of Shigella spp., Escherichia coli, and enteroinvasive E. coli (EIEC) is challenging because of their close relatedness. Distinction is vital, as infections with Shigella spp. are under surveillance of health authorities, in contrast to EIEC infections. In this study, a culture-dependent identification algorithm and a molecular identification algorithm were evaluated. Discrepancies between the two algorithms and original identification were assessed using whole-genome sequencing (WGS). After discrepancy analysis with the molecular algorithm, 100% of the evaluated isolates were identified in concordance with the original identification. However, the resolution for certain serotypes was lower than that of previously described methods and lower than that of the culture-dependent algorithm. Although the resolution of the culture-dependent algorithm is high, 100% of noninvasive E. coli, Shigella sonnei, and Shigella dysenteriae, 93% of Shigella boydii and EIEC, and 85% of Shigella flexneri isolates were identified in concordance with the original identification. Discrepancy analysis using WGS was able to confirm one of the used algorithms in four discrepant results. However, it failed to clarify three other discrepant results, as it added yet another identification. Both proposed algorithms performed well for the identification of Shigella spp. and EIEC isolates and are applicable in low-resource settings, in contrast to previously described methods that require WGS for daily diagnostics. Evaluation of the algorithms showed that both algorithms are capable of identifying Shigella species and EIEC isolates. The molecular algorithm is more applicable in clinical diagnostics for fast and accurate screening, while the culture-dependent algorithm is more suitable for reference laboratories to identify Shigella spp. and EIEC up to the serotype level.


Subject(s)
Algorithms , Dysentery, Bacillary/diagnosis , Escherichia coli Infections/diagnosis , Escherichia coli/isolation & purification , Molecular Diagnostic Techniques/standards , Shigella/isolation & purification , Bacteriological Techniques , Diagnosis, Differential , Dysentery, Bacillary/microbiology , Enterotoxigenic Escherichia coli/classification , Enterotoxigenic Escherichia coli/genetics , Enterotoxigenic Escherichia coli/isolation & purification , Escherichia coli/classification , Escherichia coli/genetics , Escherichia coli Infections/microbiology , Genes, Bacterial/genetics , Humans , Phylogeny , Serogroup , Shigella/classification , Shigella/genetics , Whole Genome Sequencing/standards
12.
J Microbiol Methods ; 131: 10-15, 2016 12.
Article in English | MEDLINE | ID: mdl-27693908

ABSTRACT

An inter-laboratory collaborative trial for the evaluation of diagnostics for detection and identification of Shigella species and Entero-invasive Escherichia coli (EIEC) was performed. Sixteen Medical Microbiological Laboratories (MMLs) participated. MMLs were interviewed about their diagnostic methods and a sample panel, consisting of DNA-extracts and spiked stool samples with different concentrations of Shigella flexneri, was provided to each MML. The results of the trial showed an enormous variety in culture-dependent and molecular diagnostic techniques currently used among MMLs. Despite the various molecular procedures, 15 out of 16 MMLs were able to detect Shigella species or EIEC in all the samples provided, showing that the diversity of methods has no effect on the qualitative detection of Shigella flexneri. In contrast to semi quantitative analysis, the minimum and maximum values per sample differed by approximately five threshold cycles (Ct-value) between the MMLs included in the study. This indicates that defining a uniform Ct-value cut-off for notification to health authorities is not advisable.


Subject(s)
Bacterial Typing Techniques/methods , Dysentery, Bacillary/diagnosis , Escherichia coli Infections/diagnosis , Escherichia coli/genetics , Molecular Diagnostic Techniques/methods , Shigella/genetics , Bacterial Typing Techniques/standards , DNA, Bacterial , Diagnosis, Differential , Diagnostic Self Evaluation , Dysentery, Bacillary/microbiology , Escherichia coli/classification , Escherichia coli/growth & development , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Feces/microbiology , Humans , Laboratories , Molecular Diagnostic Techniques/standards , Netherlands , Shigella/classification , Shigella/growth & development , Shigella/isolation & purification , Shigella flexneri/genetics , Shigella flexneri/growth & development , Shigella flexneri/isolation & purification
13.
Int J Syst Evol Microbiol ; 66(9): 3420-3425, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27259517

ABSTRACT

Bacteria were isolated from an industrial water circuit in the Netherlands. These strains were identified using API 20 NE as possible, or likely, Burkholderia pseudomallei. With VITEK 2 some of these strains scored 'low discrimination' for Francisella tularensis, amongst others. A total of twenty-six strains were assigned to the species Pseudomonas brenneri, Pseudomonas gessardii or Pseudomonas proteolytica. Because of the possibility of misidentification of these environmental species as medical- and public-health relevant B. pseudomallei and F. tularensis, an emended description, based on tests results more customarily used in clinical laboratories, was suitable. For this reason, the strains in this study, including the type strains DSM 15294T, DSM 17152T and DSM 15321T, were subjected to a polyphasic identification procedure. This procedure consisted of multiple phenotypic tests, fatty acid analysis, 16S rDNA sequence analysis, matrix-assisted laser desorption ionization time-of-flight mass spectronomy and various species-specific molecular tests. Based on the results of the polyphasic procedures, the species descriptions of P. brenneri, P. gessardii and P. proteolytica have been emended.


Subject(s)
Burkholderia pseudomallei/classification , Francisella tularensis/classification , Phylogeny , Pseudomonas/classification , Water Microbiology , Bacterial Typing Techniques , Burkholderia pseudomallei/isolation & purification , DNA, Bacterial/genetics , Fatty Acids/chemistry , Francisella tularensis/isolation & purification , Netherlands , Pseudomonas/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Species Specificity , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Water Supply
14.
Stand Genomic Sci ; 8(3): 539-47, 2013 Jul 30.
Article in English | MEDLINE | ID: mdl-24501637

ABSTRACT

Francisella tularensis is a facultative intracellular bacterium in the class Gammaproteobacteria. This strain is of interest because it is the etiologic agent of tularemia and a highly virulent category A biothreat agent. Here we describe the draft genome sequence and annotation of Francisella tularensis subsp. holarctica BD11-00177, isolated from the first case of indigenous tularemia detected in The Netherlands since 1953. Whole genome DNA sequence analysis assigned this isolate to the genomic group B.FTNF002-00, which previously has been exclusively reported from Spain, France, Italy, Switzerland and Germany. Automatic annotation of the 1,813,372 bp draft genome revealed 2,103 protein-coding and 46 RNA genes.

15.
BMC Microbiol ; 11: 267, 2011 Dec 23.
Article in English | MEDLINE | ID: mdl-22192890

ABSTRACT

BACKGROUND: The genus Brucella contains highly infectious species that are classified as biological threat agents. The timely detection and identification of the microorganism involved is essential for an effective response not only to biological warfare attacks but also to natural outbreaks. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is a rapid method for the analysis of biological samples. The advantages of this method, compared to conventional techniques, are rapidity, cost-effectiveness, accuracy and suitability for the high-throughput identification of bacteria. Discrepancies between taxonomy and genetic relatedness on the species and biovar level complicate the development of detection and identification assays. RESULTS: In this study, the accurate identification of Brucella species using MALDI-TOF-MS was achieved by constructing a Brucella reference library based on multilocus variable-number tandem repeat analysis (MLVA) data. By comparing MS-spectra from Brucella species against a custom-made MALDI-TOF-MS reference library, MALDI-TOF-MS could be used as a rapid identification method for Brucella species. In this way, 99.3% of the 152 isolates tested were identified at the species level, and B. suis biovar 1 and 2 were identified at the level of their biovar. This result demonstrates that for Brucella, even minimal genomic differences between these serovars translate to specific proteomic differences. CONCLUSIONS: MALDI-TOF-MS can be developed into a fast and reliable identification method for genetically highly related species when potential taxonomic and genetic inconsistencies are taken into consideration during the generation of the reference library.


Subject(s)
Bacterial Typing Techniques/methods , Brucella/classification , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Brucella/isolation & purification , Cluster Analysis , DNA, Bacterial/genetics , Genome, Bacterial , Minisatellite Repeats , Proteome/analysis , Species Specificity
16.
PLoS One ; 4(9): e6870, 2009 Sep 01.
Article in English | MEDLINE | ID: mdl-19721713

ABSTRACT

BACKGROUND: Caspase-12 functions as an antiinflammatory enzyme inhibiting caspase-1 and the NOD2/RIP2 pathways. Due to increased susceptibility to sepsis in individuals with functional caspase-12, an early-stop mutation leading to the loss of caspase-12 has replaced the ancient genotype in Eurasia and a significant proportion of individuals from African populations. In African-Americans, it has been shown that caspase-12 inhibits the pro-inflammatory cytokine production. METHODOLOGY/PRINCIPAL FINDINGS: We assessed whether similar mechanisms are present in African individuals, and whether evolutionary pressures due to plague may have led to the present caspase-12 genotype population frequencies. No difference in cytokine induction through the caspase-1 and/or NOD2/RIP2 pathways was observed in two independent African populations, among individuals with either an intact or absent caspase-12. In addition, stimulations with Yersinia pestis and two other species of Yersinia were preformed to investigate whether caspase-12 modulates the inflammatory reaction induced by Yersinia. We found that caspase-12 did not modulate cytokine production induced by Yersinia spp. CONCLUSIONS: Our experiments demonstrate for the first time the involvement of the NOD2/RIP2 pathway for recognition of Yersinia. However, caspase-12 does not modulate innate host defense against Y. pestis and alternative explanations for the geographical distribution of caspase-12 should be sought.


Subject(s)
Caspase 12/physiology , Gene Expression Regulation , Sepsis/metabolism , Yersinia Infections/metabolism , Yersinia pestis/pathogenicity , Cytokines/metabolism , Genotype , Humans , Immunity, Innate , Inflammation , Interleukin-1beta/metabolism , Mali , Nod2 Signaling Adaptor Protein/metabolism , Receptor-Interacting Protein Serine-Threonine Kinase 2/metabolism
17.
J Clin Microbiol ; 47(10): 3218-25, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19641058

ABSTRACT

Capnocytophaga canimorsus can be a virulent pathogen, whereas C. cynodegmi is of low virulence. Heterogeneity within these species, their frequency in dogs, and pathogenicity factors are largely unknown. Strains from blood cultures from patients presumptively identified as C. canimorsus (n = 25) and as C. cynodegmi by rrs analysis (n = 4), blood cultures from dogs (n = 8), blood cultures from cats (n = 2), and cultures from swabs from dog mouths (n = 53) were analyzed. PCR-restriction fragment length polymorphism (PCR-RFLP), a species-specific PCR on rpoB, and rrs sequencing were used. All 29 strains from human blood cultures could be grouped into three PCR-RFLP types. One included the C. canimorsus type strain, and the other types were closely related. Two canine strains were C. canimorsus and grouped into the least common RLFP pattern group. Five were C. cynodegmi and clustered with the reference strain. One canine and both feline strains were distinct. Four human strains that presumptively had been identified as C. cynodegmi by RNA gene sequence analysis clustered with the C. canimorsus strains by both PCR-RFLP and the sequence-specific PCR of the rpoB gene. C. canimorsus DNA was present in 73% (range, 61 to 85%) of dogs' mouths, and C. cynodegmi DNA was present in 96% (range, 94 to 100%) of dogs' mouths. As defined by rpoB PCR-RFLP and by PCRs using specific primers, all strains from human blood were C. canimorsus. The sequencing of rrs genes suggested the presence of different gene copies in a few strains, indicating that the method is less appropriate for species identification. Both species are present in the majority of dogs. Additional Capnocytophaga species occur in dogs' and cats' mouths.


Subject(s)
Capnocytophaga/classification , Capnocytophaga/isolation & purification , Cat Diseases/microbiology , Dog Diseases/microbiology , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/veterinary , Polymerase Chain Reaction/methods , Animals , Blood/microbiology , Capnocytophaga/genetics , Cat Diseases/epidemiology , Cats , Cluster Analysis , DNA Fingerprinting , DNA, Bacterial/genetics , DNA-Directed RNA Polymerases/genetics , Dog Diseases/epidemiology , Dogs , Female , Gram-Negative Bacterial Infections/epidemiology , Humans , Male , Molecular Sequence Data , Polymorphism, Restriction Fragment Length , Prevalence , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
18.
Vet Microbiol ; 128(3-4): 395-9, 2008 Apr 30.
Article in English | MEDLINE | ID: mdl-18023543

ABSTRACT

Streptobacillus moniliformis is a zoonotic bacterium. We obtained positive S. moniliformis PCR results in oral swab samples from guinea pigs from an experimental colony and the breeding colony of origin. Comparison of the DNA sequence of an amplicon with deposited 16S rDNA sequences revealed that Leptotrichia sp. can be the source of a false positive S. moniliformis PCR outcome.


Subject(s)
Fusobacterium Infections/veterinary , Guinea Pigs/microbiology , Leptotrichia/isolation & purification , Polymerase Chain Reaction/veterinary , Rodent Diseases/diagnosis , Streptobacillus/isolation & purification , Animals , Base Sequence , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , False Positive Reactions , Fusobacterium Infections/diagnosis , Fusobacterium Infections/microbiology , Molecular Sequence Data , Polymerase Chain Reaction/methods , Polymerase Chain Reaction/standards , Rodent Diseases/microbiology , Sequence Alignment/veterinary , Species Specificity
19.
Antonie Van Leeuwenhoek ; 83(2): 135-48, 2003.
Article in English | MEDLINE | ID: mdl-12785307

ABSTRACT

Intestinal microorganisms play an important role in plant fiber degradation by larvae of the rose chafer Pachnoda marginata. In the hindgut of the larvae 2.5 to 7.4 x 10(8) bacteria per ml of gut content with xylanase or endoglucanase activity were found. Bacteria in the midgut were not (hemi)cellulolytic, but the alkaline environment in this part of the intestinal tract functions as a precellulolytic phase, solubilizing part of the lignocellulosic material. Accordingly, the degradation of lignocellulose-rich material in Pachnoda marginata larvae appeared to be a combination of a physico-chemical and microbiological process. A number of different facultative anaerobic and strictly anaerobic bacteria with (hemi)cellulolytic activity were isolated from the hindgut. A dominant (hemi)cellulolytic species was a Gram positive, irregular shaped, facultative aerobic bacterium. Further physiological identification placed the isolate in the genus Promicromonospora. Comparative 16S rDNA analysis and phenotypic features revealed that the isolate represented a new species for which the name Promicromonospora pachnodae is proposed. P. pachnodae produced xylanases and endoglucanases on several plant derived polymers, both under aerobic and anaerobic conditions.


Subject(s)
Actinomycetales/classification , Actinomycetales/isolation & purification , Cellulase , Cellulose/metabolism , Coleoptera/microbiology , Intestines/microbiology , Actinomycetales/genetics , Actinomycetales/growth & development , Animals , Bacterial Typing Techniques , Coleoptera/growth & development , DNA, Ribosomal/analysis , Fatty Acids/analysis , Glycoside Hydrolases/metabolism , Larva/microbiology , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Xylan Endo-1,3-beta-Xylosidase , Xylosidases/metabolism
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