Brucellosis is a zoonosis with non-specific clinical symptoms involving multiple systems and organs. Its prevalence is low in most of EU countries, which can lead to the difficulties in laboratory and clinical diagnostic. Due to its relationship to the Ochrobactrum spp., it may be misclassified in rapid identification systems. We present a case of a 13-year-old immunocompetent girl who was examined several times for fever, fatigue, night sweats and weight loss; laboratory results showed mildly elevated C-reactive protein, anaemia and leukopenia. Four weeks before the onset of symptoms, she had been on a family holiday in Egypt. Given her symptoms, a haemato-oncological or autoimmune disease was considered more likely. The diagnosis of Brucella spondylitis was made after 4 months. The main reasons for this delay were as follows: low specificity of clinical symptoms, delay in completing the travel history, inconclusive initial serological results and misidentification of the blood culture isolate as Ochrobactrum sp. Even in countries with a low incidence of brucellosis, it is essential to educate healthcare professionals about the disease. Low specificity of symptoms and limited experience of laboratory staff may lead to late diagnosis with risk of complications and poor outcome. If Ochrobactrum spp. is detected in clinical specimens by rapid identification, careful re-evaluation must follow and all measures to prevent laboratory-acquired infections must be taken until Brucella spp. is unequivocally excluded.
- MeSH
- bakteriemie * diagnóza mikrobiologie MeSH
- Brucella izolace a purifikace klasifikace MeSH
- brucelóza * diagnóza mikrobiologie MeSH
- chybná diagnóza * MeSH
- gramnegativní bakteriální infekce diagnóza mikrobiologie MeSH
- horečka * mikrobiologie etiologie MeSH
- lidé MeSH
- mladiství MeSH
- Ochrobactrum * genetika izolace a purifikace MeSH
- spondylitida mikrobiologie diagnóza MeSH
- Check Tag
- lidé MeSH
- mladiství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- Geografické názvy
- Egypt MeSH
Brucellosis is one of the most important zoonoses worldwide, primarily affecting livestock but also posing a serious threat to public health. The major Brucella species are known to cause a feverish disease in humans with various clinical signs. These classical Brucella species are (re-)emerging, but also novel strains and species, some of them transmitted from rodents, can be associated with human infections. As a result of our review on rodent-borne brucellosis, we emphasise the need for more comprehensive surveillance of Brucella and especially Brucella microti in rodent populations and call for further research targeting the ecological persistence of rodent-associated Brucella species in the environment, their epizootic role in wild rodents and their virulence and pathogenicity for wildlife.
- MeSH
- Brucella * izolace a purifikace MeSH
- brucelóza * epidemiologie veterinární mikrobiologie přenos MeSH
- hlodavci * mikrobiologie MeSH
- lidé MeSH
- veřejné zdravotnictví * MeSH
- zoonózy * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Brucellosis is a zoonosis caused by Brucella, which poses a great threat to human health and animal husbandry. Pathogen surveillance is an important measure to prevent brucellosis, but the traditional method is time-consuming and not suitable for field applications. In this study, a recombinase polymerase amplification-SYBR Green I (RPAS) assay was developed for the rapid and visualized detection of Brucella in the field by targeting BCSP31 gene, a conserved marker. The method was highly specific without any cross-reactivity with other common bacteria and its detection limit was 2.14 × 104 CFU/mL or g of Brucella at 40 °C for 20 min. It obviates the need for costly instrumentation and exhibits robustness towards background interference in serum, meat, and milk samples. In summary, the RPAS assay is a rapid, visually intuitive, and user-friendly detection that is highly suitable for use in resource-limited settings. Its simplicity and ease of use enable swift on-site detection of Brucella, thereby facilitating timely implementation of preventive measures.
- MeSH
- Brucella * genetika izolace a purifikace MeSH
- brucelóza * diagnóza mikrobiologie MeSH
- DNA bakterií genetika MeSH
- lidé MeSH
- limita detekce MeSH
- mléko mikrobiologie MeSH
- rekombinasy * metabolismus genetika MeSH
- senzitivita a specificita MeSH
- skot MeSH
- techniky amplifikace nukleových kyselin * metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Background:Brucella microti is a pathogen of rodents and wild mammals. Here, we report the first probable infection with B. microti in a mammalogist. Materials and Methods: In the study, we provided complete clinical description as well as laboratory analysis of probable human infection caused by B. microti. Results: Considering the clinical course of the infection, the obvious epidemiological link (a bite by an infected rodent), the isolation of a pathogen from a sick vole that was affected by clinical infection with B. microti, and the specific serological response (slow agglutination test) in human patient, we can conclude that the human disease described here was probably caused by B. microti, an emerging bacterial pathogen transmitted by rodents. Conclusion: Rodents and other wildlife need to be monitored not only for established zoonotic agents such as hantaviruses, lymphocytic choriomeningitis virus, Leptospira spp., Francisella tularensis, but also for Brucella microti and other atypical rodent-borne brucellae.
- MeSH
- Arvicolinae mikrobiologie MeSH
- Bacteria MeSH
- Brucella * MeSH
- divoká zvířata MeSH
- hlodavci MeSH
- lidé MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In the present study, bacterial isolates were screened for arsenic resistance efficiency. Environmental isolates were isolated from arsenic-rich soil samples (i.e., from Rajnandgaon district of Chhattisgarh state, India). Amplification and sequencing of 16S rRNA gene revealed that the isolates were of Bacillus firmus RSN1, Brevibacterium senegalense RSN2, Enterobacter cloacae RSN3, Stenotrophomonas pavanii RSN6, Achromobacter mucicolens RSN7, and Ochrobactrum intermedium RSN10. Arsenite efflux gene (arsB) was successfully amplified in E. cloacae RSN3. Atomic absorption spectroscopy (AAS) analysis showed an absorption of 32.22% arsenic by the RSN3 strain. Furthermore, results of scanning electron microscopy (SEM) for morphological variations revealed an initial increase in the cell size at 1 mM sodium arsenate; however, it was decreased at 10 mM concentration in comparison to control. This change of the cell size in different metal concentrations was due to the uptake and expulsion of the metal from the cell, which also confirmed the arsenite efflux system.
Due to limitations in commercial diagnostic methods, this study aimed to develop a reliable real-time polymerase chain reaction (Rt-PCR) assay for early diagnosis of brucellosis. Optimization of the Rt-PCR method was performed on serum samples spiked by Brucella melitensis with different densities ranging from 101 to 108 colony-forming units (cfu)/mL; each density was prepared in ten samples. The limit of detection was investigated by using Thermo DNA extraction kit with Maxima SYBR Green Rt-PCR and two TaqMan probe-based Rt-PCR protocols performed by QuantiTect and TEMPase multiplex PCR master mixes in two thermal cyclers, which were Rotor-Gene and Bio-Rad. The validation of the optimized protocol was carried on 20 brucellosis-negative samples and 20 samples spiked with B. melitensis by using a combination of Thermo DNA extraction kit with TEMPase PCR master mix. SYBR Green Rt-PCR yielded positive results on all samples having ≥ 104 cfu/mL of B. melitensis in both thermal cyclers. Its limit of detection was 112 DNA copies per reaction. The positivity of both probe-based Rt-PCR protocols was 100% and 80% on the samples having 103 cfu/mL and 102 cfu/mL of B. melitensis, respectively. The limit of detection of probe-based protocols was defined as 4 DNA copies per reaction. The optimized Rt-PCR protocol showed high-level accuracy, precision, specificity, and sensitivity, each having a rate of 100%. The current study indicated that the TaqMan probe-based Rt-PCR protocol optimized and validated with serum samples can be reliably used for early diagnosis of brucellosis.
The evolutionary "success" of the genus Brucella depends on the ability to persist both in the environment as well as inside of even activated macrophages of the animal host. For that, the Brucellae produce catalase and superoxide dismutase to defend against oxidative stress. Since the deletion of the mglA gene in the B. abortus S19 vaccine strain resulted not only in an increased tolerance to H2O2 but also in the induction of cytokines in macrophages, we here investigated the effect of oxidative stress (Fe2+ and H2O2) on the survival of B. abortus S19 and the isogenic B. abortus S 19 ∆mglA 3.14 deletion mutant in comparison with B. neotomae 5K33, Brucella strain 83/13, and B. microti CCM4915. These Brucellae belong to different phylogenetic clades and show characteristic differences in the mgl-operon. From the various Brucellae tested, B. abortus S19 showed the highest susceptibility to oxidative stress and the lowest ability to survive inside of murine macrophages. B. abortus S19 ∆mglA 3.14 as well as B. neotomae, which also belongs to the classical core clade of Brucella and lacks the regulators of the mgl-operon, presented the highest degree of tolerance to H2O2 but not in the survival in macrophages. The latter was most pronounced in case of an infection with B. 83/13 and B. microti CCM4915. The various Brucellae investigated here demonstrate significant differences in tolerance against oxidative stress and different survival in murine macrophages, which, however, do not correlate directly.
- MeSH
- adenosintrifosfát metabolismus MeSH
- bakteriální geny MeSH
- Brucella abortus fyziologie MeSH
- Brucella klasifikace fyziologie MeSH
- buněčné linie MeSH
- cytokiny metabolismus MeSH
- druhová specificita MeSH
- makrofágy imunologie mikrobiologie MeSH
- mikrobiální viabilita MeSH
- mutace MeSH
- myši MeSH
- oxidační stres * MeSH
- peroxid vodíku metabolismus MeSH
- počet mikrobiálních kolonií MeSH
- železo metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
As dendritic cells (DCs) are among the first cells to encounter antigens, these cells trigger both innate and T cell responses, and are the most potent antigen-presenting cells. Brucella spp., which is an intracellular facultative and stealthy pathogen, is able to evade the bactericidal activities of professional phagocytes. Several studies have demonstrated that Brucella can survive and replicate intracellularly, thereby provoking impaired maturation of DCs. Therefore, the interaction between DCs and Brucella becomes an interesting model to study the immune response. In this review, we first will describe the most common techniques for DCs differentiation in vitro as well as general features of brucellosis. Then, the interaction of DCs and Brucella, including pathogen recognition, molecular mechanisms of bacterial pathogenesis, and intracellular trafficking of Brucella to subvert innate response, will be reviewed. Finally, we will debate diversity in immunological DC response and the controversial role of DC activation against Brucella infection.
- MeSH
- Brucella imunologie patogenita MeSH
- brucelóza imunologie MeSH
- cytoplazma mikrobiologie MeSH
- dendritické buňky mikrobiologie MeSH
- interakce hostitele a patogenu imunologie MeSH
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
The aim of this work was to compare production of endotoxin and to determine susceptibility to antibiotics in two groups of specimens-wild-type strains Ochrobactrum anthropi isolated from the environment and the strains isolated from patients with cystic fibrosis. The determination of the endotoxin produced by the test strains was carried on by using a limulus amebocyte lysate test (LAL test). Determination of ATB sensitivity was accomplished by means of a broth dilution method in a microtiter plate (MIC). No significant difference was found between the group of ochrobacters isolated from the environment and the group of ochrobacters isolated from cystic fibrosis patients. Antibiotic sensitivity testing has indicated that the resistance to tigecycline, trimethoprim/sulfamethoxazole, and gentamicin was slightly higher in strains isolated from cystic fibrosis patients in comparison with strains isolated from the environment. In general, most of the test strains were sensitive to most of the antibiotics tested. Significant resistance has been demonstrated for cefotaxime. Resistance was also found for gentamicin in strains number 4 and 7. The MIC was equal to the breakpoint for this antibiotic (8000 mg/L).
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence MeSH
- cystická fibróza mikrobiologie MeSH
- dítě MeSH
- endotoxiny metabolismus MeSH
- gramnegativní bakteriální infekce mikrobiologie MeSH
- lidé MeSH
- Limulus test MeSH
- mikrobiální testy citlivosti MeSH
- mikrobiologie životního prostředí MeSH
- mladiství MeSH
- Ochrobactrum anthropi účinky léků izolace a purifikace metabolismus MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
Brucella canis infection is a neglected zoonotic disease and its seroprevalence in dogs and at-risk humans has not been previously studied in several countries including Jordan. The main aim of this study was to determine the seroprevalence and identify risk factors of B. canis infection in police, breeding and stray dogs and in at-risk humans in Jordan. A total of 169 sera samples from apparently healthy dogs and 185 samples from apparently healthy people (85 from dog handlers and 100 from the general population) were tested in the study. Antibodies against B. canis were tested using the canine D-Tec® CB Rapid Slide Agglutination Test (RSAT) kit with secondary 2-mercaptoethanol (ME-RSAT). Overall, 8.3% of the dog sera samples tested positive to antibodies against B. canis, and 37.8% of stray dogs tested positive. Seroprevalence was higher in male dogs than in females. Furthermore, none of the tested human samples was positive to antibodies against B. canis. There was a significant association between seropositivity and the type of dog. The study reports preliminary findings that suggest the presence of B. canis among stray dogs in Jordan. Thus, preventive measures should be taken to control the transmission of this pathogen from stray dogs to other dogs and humans as well.
