Comparative study of cefixime and tetracycline as an evaluation policy driven by the antibiotic resistance crisis in Indonesia.

Antibiotic resistance is a serious threat that occurs globally in the health sector due to increased consumption of inappropriate antibiotics. Guidelines for prescribing antibiotics for ARTIs have been issued in general practice to promote rational antibiotic prescribing. This study was conducted to compare the effectiveness of cefixime and tetracycline as a solution to improve monitoring of appropriate antibiotic use in the treatment of ARTIs. All stock isolates were rejuvenated first, and cultured on standard media and Kirby-Bauer disc diffusion method was used for susceptibility testing in accordance with the Clinical and Laboratory Standard Institute's (CLSI) recommendations. Identification of bacteria from a single isolate was carried out to determine which bacteria were resistant to cefixime and tetracycline. A total of 466 single isolates of bacteria were analyzed, which showed a percentage of resistance to cefixime 38.0%, and tetracycline 92.86%. Bacterial isolates were resistant to cefixime and tetracycilne was a genus of Haemophilus, Streptococcus, Corynebacterium, Staphylococcus, and bordetella. Cefixime compared to tetracycline was proven to be superior in terms of the effectiveness of ARIs treatment.

Isolation of Bordetella trematum from the respiratory tract of a patient with lung cancer: a case report.

We report the case of isolation of Bordetella trematum from the respiratory tract of a patient with lung carcinoma. This gram-negative, opportunistic rod was firstly described in 1996. To date, only several strains of Bordetella trematum have been isolated and reported, mostly from skin and soft tissue infections. The patient was admitted to the ICU of the Pulmonary Department in incipient septic shock with respiratory failure. Intravenous fluid resuscitation and non-invasive ventilation were administered immediately. A broad spectrum antibiotic piperacillin/tazobactam was administered empirically after sampling of material for microbiological examination. The bronchoscopy showed a large cavern of decayed tumour invading into mediastinum. Both sample cultures showed significant quantities of gram-negative non-fermenting bacteria. The isolate was identified using MALDI-TOF MS as Bordetella trematum and the identification was confirmed using 16S ribosomal RNA sequencing. In the last few years, routine bacterial identification using MALDI-TOF MS has enabled correct discrimination of this species. Nevertheless, isolation of Bordetella trematum in clinical samples is still very uncommon, and it is appropriate to confirm the species identification via 16S ribosomal RNA sequencing. To our knowledge, this is the first case of B. trematum isolated from the human respiratory tract since its first description. The clinical significance of Bordetella trematum in the rapid deterioration of the patient's status remains unclear.

Spondylodiscitis caused by Bordetella holmesii, a misrecognized pathogen emerging in invasive infections.

We report a case of spondylodiscitis caused by Bordetella holmesii, an emergent pathogen. This small Gram-negative rod was first known as a cause of invasive infections on asplenic patients. This case describes a spondylodiscitis due to this bacterium in an immunocompetent patient. This article underlines the interest of prolonged incubation for specimens in case of spondylodiscitis and shows us the contributions of mass spectrometry for easy and rapid identification of such bacterium.

Cooperativity between antibiotics and antiseptics: testing the bactericidal effect.

OBJECTIVE: Treatment with antibiotics together with local application of antiseptics is common in wound care. We investigated the effectiveness of an antiseptic in two variations: octenidine (Oct) and octenidine+ (Oct+ with isotonic glucose addition). METHOD: Using the agar diffusion test with cultures of pathogenic Staphylococcus aureus and the non-pathogenic Bordetella petrii, we compared the effectiveness of octenidine to the classical antiseptics beta-isodona (povidone-iodine; PI), chlorhexidine (Chl) and taurolin (Tau) alone, and in combination with various common antibiotics to uncover cooperativity between antiseptics and antibiotics. RESULTS: We detected strong interactions between antibiotics and antiseptics, that either enhanced or reduced the bactericidal efficiency. Effectiveness was dependent on the type of organism tested. Oct applied together with ineffective antibiotics frequently led to effective growth inhibition of Bordetella petrii. With Staphylococcus aureus we did not find such an effect. To this end, we reason that positively charged Oct may associate with antibiotic compounds via electrostatic interactions and guide it more efficiently to the bacterial cell wall. Interaction with antibiotics sometimes led to sequestration and reduced availability of some antiseptic/antibiotic combinations, but never with Oct. CONCLUSION: These data provide new arguments for decision planning concerning the choice of agent in the treatment of wound infections.

Bordetella bronchiseptica infection.

OBJECTIVE: To collect data of all patients admitted to hospital with a positive test to Bordetella bronchiseptica between 2001 and 2015. METHODS: We performed a retrospective monocentric study of all hospitalized patients over the past 15 years with a positive test to B. bronchiseptica. RESULTS: Nine patients were included between 2001 and 2015; two presented with infectious relapses, i.e. a total of 14 positive test samples were observed. Age, induced immunodeficiency, and preexisting respiratory illnesses are risk factors. All patients showed symptoms at sample collection and the infection was exclusively respiratory. The diagnosis was obtained through a cytobacteriological test of sputum, bronchial aspiration, or bronchial fibroscopy with a bronchoalveolar lavage. The drug susceptibility test revealed a natural resistance to cephalosporins including ceftazidime, monobactam, and fosfomycin. There were cases of resistance to penicillin A and to the trimethoprim/sulfamethoxazole association. The classically used antibiotic treatment for community-acquired pneumonia is based on probability and may thus fail. Four patients died. The duration and nature of the antibiotics to use have not been codified. CONCLUSION: B. bronchiseptica infection mainly affects the elderly. All patients should be treated, regardless of the importance of the inoculum, and all infected animals should be treated.

Bordetella pseudohinzii as a Confounding Organism in Murine Models of Pulmonary Disease.

A group studying acute lung injury observed an increased percentage of neutrophils in the bronchoalveolar lavage (BAL) fluid of mice. BAL was performed, and lung samples were collected sterilely from 5 C57BL/6 mice that had been bred inhouse. Pure colonies of bacteria, initially identified as Bordetella hinzii were cultured from 2 of the 5 mice which had the highest percentages of neutrophils (21% and 26%) in the BAL fluid. Subsequent sequencing of a portion of the ompA gene from this isolate demonstrated 100% homology with the published B. pseudohinzii sequence. We then selected 10 mice from the investigator's colony to determine the best test to screen for B. pseudohinzii in the facility. BAL was performed, the left lung lobe was collected for culture and PCR analysis, the right lung lobe and nasal passages were collected for histopathology, an oral swab was collected for culture, and an oral swab and fecal pellets were collected for PCR analysis. B. pseudohinzii was cultured from the oral cavity, lung, or both in 8 of the 10 mice analyzed. All 8 of these mice were fecal PCR positive for B. pseudohinzii; 7 had increased neutrophils (5% to 20%) in the BAL fluid, whereas the 8th mouse had a normal neutrophil percentage (2%). Active bronchopneumonia was not observed, but some infected mice had mild to moderate rhinitis. B. pseudohinzii appears to be a microbial agent of importance in mouse colonies that can confound pulmonary research. Commercial vendors and institutions should consider colony screening, routine reporting, and exclusion of B. pseudohinzii.

Bordetella trematum sepsis with shock in a diabetic patient with rapidly developing soft tissue infection.

Bordetella is a gram-negative, glucose non-fermenting bacillus, consisting of many host-associated species. B. trematum has previously been identified in wound infections, but rarely known to be a source of bacteremia. Currently, 16S rRNA sequencing represents the reference standard method by which identification is made. Herein, we present a case of fatal B. trematum bacteremia with septic shock. The presumed primary site of the infection was a rapidly developing left leg deep soft tissue infection without necrotizing fasciitis. B. trematum should now be considered as a significant pathogen in sepsis.

Bordetella holmesii: Still Emerging and Elusive 20 Years On.

Since the first description of Bordetella holmesii in 1995, almost 100 publications have contributed to the increasing knowledge of this emerging bacterium. Although first reported to induce bacteremia mainly in immunocompromised patients, it has also been isolated in healthy persons and has shown the capacity to induce pertussis-like symptoms and other clinical entities, such as meningitis, arthritis, or endocarditis. Respiratory diseases are generally less severe than those induced by Bordetella pertussis. However, B. holmesii was found to have a higher capacity of invasiveness given the various infection sites in which it was isolated. The diagnosis is difficult, particularly as it is a slow-growing organism but also because respiratory infections are systematically misdiagnosed as B. pertussis. Treatment is delicate, as its susceptibility to macrolides (prescribed in respiratory infections) and ceftriaxone (used in invasive disease) is challenged. Regarding prevention, there is no consensus on prophylactic treatment following index cases and no vaccine is available. Epidemiological data are also sparse, with few prevalence studies available. In this chapter, we provide an overview of the current state of knowledge on B. holmesii.

Molecular and antimicrobial analyses of non-classical Bordetella isolated from a laboratory mouse.

Accurate identification and separation of non-classical Bordetella species is very difficult. These species have been implicated in animal infections. B. hinzii, a non-classical Bordetella, has been isolated from mice in experimental facilities recently. We isolated and characterized one non-classical Bordetella isolate from the trachea and lung of an ICR mouse. Isolate BH370 was initially identified as B. hinzii by 16S ribosomal DNA and ompA sequencing. Additionally, isolate BH370 also displayed antimicrobial sensitivity profiles similar to B. hinzii. However, analyses of nrdA sequences determined its identity as Bordetella genogroup 16. The isolation of BH370 from a healthy mouse suggests the possibility of it being a commensal. The nrdA gene was demonstrated to possess greater phylogenetic resolution as compared with 16S ribosomal DNA and ompA for the discrimination of non-classical Bordetella species.

Occurrence of 3 Bordetella species during an outbreak of cough illness in Ohio: epidemiology, clinical features, laboratory findings and antimicrobial susceptibility.

BACKGROUND: An increase in laboratory diagnosis of pertussis was noted in central Ohio during 2010. Diagnosis was made using a polymerase chain reaction assay targeting the multicopy insertion sequence IS481, which is found in both Bordetella pertussis (Bp) and Bordetella holmesii (Bh). An increase in specimens testing positive for Bordetella parapertussis (Bpp) using insertion sequence IS1001 was also noted. METHODS: Nasopharyngeal swab specimens submitted April 1, 2010, to March 31, 2011, were tested using a multiplex polymerase chain reaction assay for Bp/Bh (IS481) and Bpp followed by singleplex assays for Bp and Bh. A subgroup of specimens was also cultured for Bordetella species, and antimicrobial susceptibility testing was performed on recovered organisms. Demographic and clinical features were compared for patients with Bp, Bh and Bpp. RESULTS: Of 520 IS481-positive specimens, 214 (41.1%) were positive for Bp, 79 (15.2%) were positive for Bh and 5 (1.0%) were positive for both Bp and Bh; 222 (42.7%) were negative for both targets. An additional 220 specimens were positive for Bpp. Among a sample of 155 IS481-positive specimens, 40, 15 and 0 were culture positive for Bp, Bh and Bpp, respectively. Among a sample of 55 BparaIS1001-positive (Bpp) specimens, 22, 0 and 0 were culture positive for Bpp, Bp and Bh, respectively. All Bordetella species were susceptible to macrolide antibiotics. Patients with Bh were older than patients with Bp, who were older than those positive for Bpp (mean ages: 12.0, 8.0 and 4.2 years, respectively; P < 0.001). One or more classic signs of pertussis (ie, paroxysmal cough, whoop, post-tussive emesis) were seen in 55.9% of 263 patients (59 Bp, 24 Bh, 80 Bpp and 100 negative for Bordetella species), but did not differ statistically among the groups (χ = 5.1, P = 0.17). CONCLUSIONS: All 3 Bordetella species, Bp, Bh and Bpp, were detected during on outbreak of pertussis-like cough illness. There were noted differences in age and seasonality, but clinical features at the time of presentation did not allow clear differentiation of these infections. All Bordetella species recovered from culture and tested were susceptible in vitro to macrolide antibiotics. Additional study is necessary to further characterize epidemiologic and clinical characteristics of Bh-associated cough illness and to determine potential co-occurrence of Bordetella species with other bacterial and viral respiratory tract pathogens.

Bronchitis caused by Bordetella holmesii in a child with asthma misdiagnosed as mycoplasmal infection.

We report a case of a bronchitis caused by Bordetella holmesii in a 2-year-old girl with asthma. The patient had a moderate fever and productive cough, and her condition was initially diagnosed as mycoplasmal bronchitis on the basis of her clinical symptoms and rapid serodiagnosis of mycoplasmal infection. She was treated with a bronchodilator and clarithromycin, which resulted in complete recovery. However, after the initial diagnosis, nucleic acid amplification tests of her sputum showed the absence of both Mycoplasma pneumoniae and Bordetella pertussis infections. Sputum culture showed the presence of a slow-growing, gram-negative bacillus in pure culture on Bordetella agar plates; the bacillus was later identified as B. holmesii. B. holmesii infection is rare in immunocompetent children; however, the organism is a true pathogen that can cause bronchitis in young children with asthma.

Bordetella holmesii meningitis in a 12-year-old anorectic girl.

We describe a 12-year-old anorectic girl with Bordetella holmesii meningitis, the techniques used for its identification, and minimum inhibitory concentrations of antibiotics for 7 B. Holmesii strains collected in the Netherlands during the past 12 years. B. holmesii meningitis has not been previously reported.

Bordetella petrii infection with long-lasting persistence in human.

We report the repeated isolation of Bordetella petrii in the sputum of a 79-year-old female patient with diffuse bronchiectasis and persistence of the bacterium for >1 year. The patient was first hospitalized due to dyspnea, which developed into severe cough with purulent sputum that yielded B. petrii on culture. After this first episode, the patient was hospitalized an additional 4 times with bronchorrhea symptoms. The isolates collected were analyzed by using biochemical, genotypic, and proteomic tools. Expression of specific proteins was analyzed by using serum samples from the patient. The B. petrii isolates were compared with other B. petrii isolates collected from humans or the environment and with isolates of B. pertussis, B. parapertussis, B. bronchiseptica, and B. holmesii, obtained from human respiratory tract infections. Our observations indicate that B. petrii can persist in persons with chronic pulmonary obstructive disease as has been previously demonstrated for B. bronchiseptica.

The Bordetella type III secretion system effector BteA contains a conserved N-terminal motif that guides bacterial virulence factors to lipid rafts.

The Bordetella type III secretion system (T3SS) effector protein BteA is necessary and sufficient for rapid cytotoxicity in a wide range of mammalian cells. We show that BteA is highly conserved and functionally interchangeable between Bordetella bronchiseptica, Bordetella pertussis and Bordetella parapertussis. The identification of BteA sequences required for cytotoxicity allowed the construction of non-cytotoxic mutants for localization studies. BteA derivatives were targeted to lipid rafts and showed clear colocalization with cortical actin, ezrin and the lipid raft marker GM1. We hypothesized that BteA associates with the cytoplasmic face of lipid rafts to locally modulate host cell responses to Bordetella attachment. B. bronchiseptica adhered to host cells almost exclusively to GM1-enriched lipid raft microdomains and BteA colocalized to these same sites following T3SS-mediated translocation. Disruption of lipid rafts with methyl-beta-cyclodextrin protected cells from T3SS-induced cytotoxicity. Localization to lipid rafts was mediated by a 130-amino-acid lipid raft targeting domain at the N-terminus of BteA, and homologous domains were identified in virulence factors from other bacterial species. Lipid raft targeting sequences from a T3SS effector (Plu4750) and an RTX-type toxin (Plu3217) from Photorhabdus luminescens directed fusion proteins to lipid rafts in a manner identical to the N-terminus of BteA.

Isolation and identification of a novel cis-epoxysuccinate hydrolase-producing Bordetella sp. BK-52 and optimization of enzyme production.

OBJECTIVE: To isolate and identify a novel strain with cis-epoxysuccinate hydrolase (CESH) activity and to optimize its enzyme production. METHODS: The isolated strain was identified by electron microscopy, Biolog gram negative (GN) test, G+C (guanine plus cytosine) content measurement and 16S rDNA sequence. The purified enzymatic biotransformed product was identified by IR, 1H-NMR, 13C-NMR, MS and optical rotation analysis. Then the fermentation conditions for CESH production were optimized. RESULTS: A novel CESH-producing strain was isolated for biotransforming cis-epoxysuccinate to D(-)-tartaric acid. It was assigned to genus Bordetella and named Bordetella sp. BK-52. The optimal conditions were found to be 30 degrees C, pH 7.0, fermentation time 36 h, carbon source of saccharose, inorganic nitrogen source of ammonium sulfate and enzyme inducer of disodium cis-epoxysuccinate. Under these conditions, the maximum CESH activity reached 764 U/g biomass. CONCLUSION: The isolated Bordetella sp. BK-52 provided a new alternative for biosynthesis of D(-)-tartaric acid from cis-epoxysuccinate.

A UK clinical isolate of Bordetella hinzii from a patient with myelodysplastic syndrome.

What is believed to be the first clinical isolate of Bordetella hinzii in the UK, from a patient with myelodysplastic syndrome, is described. This patient had no known avian exposure, and the source of the organism remains unknown. It appears that the underlying immune deficiency of the patient increased the susceptibility to opportunistic infection with this organism. Human infection with B. hinzii is rare and this species is difficult to differentiate from Bordetella avium by routine phenotypic methods. Confirmation can be reliably achieved using genotypic methods, and the greater mutational variation of the ompA gene compared to other genes (e.g. 16S rRNA gene) allows unambiguous identification of this and other non-classical Bordetella species.

Bordetella petrii from a clinical sample in Australia: isolation and molecular identification.

The first isolation of Bordetella petrii from a patient with chronic suppurative mastoiditis is reported. Molecular characterization of the isolate was performed by sequencing the small-subunit rRNA gene, the Bordetella outer-membrane protein A gene (ompA) and the RisA response regulator gene (risA). This is the first reported case of B. petrii causing suppurative mastoiditis and only the second documented case of a clinically significant B. petrii isolate.

Bioremediation of toluene and naphthalene: development and validation of a GC-FID method for their monitoring.

Bioremediation of toluene and naphthalene in liquid cultures of bacteria grown in the presence of these aromatic compounds as unique sources of carbon was investigated by gas chromatography (GC). For this purpose, a method based on the use of GC with flame ionization detection was developed and validated. Validation was carried out in terms of limit of detection (LOD), limit of quantitation (LOQ), linearity, precision and trueness. In the case of naphthalene, LOD and LOQ values of 0.43 and 0.72 mg kg(-1) were achieved. Linearity was established over one order of magnitude in the range of interest, i.e. 10-100 mg kg(-1). Excellent precision was obtained both in terms of intra-day repeatability and between-day precision on two concentration levels (RSD% lower than 0.5%). A recovery of 97.9 +/- 0.2% (n=3) was calculated by addition of 640 mg kg(-1) of naphthalene to the Bushnell & Haas mineral salts basal solution containing the micro-organisms. Findings clearly showed a reduction of the naphthalene content equal to 50% and 75% after two and four weeks of contact with the micro-organisms, whereas a lower degradation was shown in the case of toluene. Finally bioremediation activity was ascribed to two different microbial populations, Bordetella Petrii and Bacillus Sphericus, which survived in the polluted medium.