A new spray-based method for the in-vitro development of dry-surface biofilms.

The inanimate environment immediately surrounding the patient in healthcare facilities is a reservoir of microorganisms embedded in dry-surface biofilms (DSB). These biofilms, first highlighted in 2012, are increasingly studied, but currently available in-vitro models only allow for the growth of semi-hydrated biofilms. We developed a new in-vitro method under actual dehydration conditions based on the hypothesis that surface contamination is mainly due to splashes of respiratory secretions. The main objective of this study was to show that the operating conditions we have defined allowed the growth of DSB with a methicillin resistant Staphylococcus aureus strain. The second objective was to show that extended-spectrum beta-lactamase-producing Enterobacteriaceae, that is, Klebsiella pneumoniae and Enterobacter cloacae were also able to grow such biofilms under these conditions. Monobacterial suspensions in sterile artificial saliva (SAS) were sprayed onto polyethylene surfaces. Nutrients and hydration were provided daily by spraying SAS enriched with 20% of Brain Heart Infusion broth. The primary outcome was mean surface coverage measured by image analysis after crystal violet staining. The method applied to S. aureus for 12 days resulted in reproducible and repeatable DSB consisting of isolated and confluent microcolonies embedded in extracellular polymeric substances as shown in scanning electron microscopy images. Similar DSB were obtained with both Enterobacteriaceae applying the same method. No interspecies variation was shown between the three strains in terms of surface coverage. These first trials are the starting point for a 3-year study currently in process.

COVID-19 and Pasteurella multocida Pulmonary Coinfection: A Case Series.

OBJECTIVES: In COVID-19 patients, bacterial and fungal pulmonary coinfections, such as Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, or Aspergillus, have been reported, but to our knowledge, no case has been reported due to Pasteurella multocida. PATIENTS AND METHODS: We describe three cases of Pasteurella multocida coinfections occurring during the 4th wave of COVID-19 in Martinique (French West Indies). RESULTS: All three cases were fatal; thus, Pasteurella multocida has to be considered as a potentially severe coinfection agent. CONCLUSIONS: Alteration of the epithelial-endothelial barrier due to a SARS-CoV-2 infection probably promotes the expression of a Pasteurella infection. In addition, the SARS-CoV-2 infection induced immunosuppression, and an inflammatory cascade could explain the infection's severity. The use of corticosteroids, which are part of the first-line therapeutic arsenal against COVID-19, may also promote the pathogenicity of this agent.

Case Report: Pulmonary Leptospirosis Misdiagnosed as COVID-19.

We report the case of an 83-year-old woman with acute, febrile respiratory failure resulting from interstitial pneumonia that required high-flow oxygen therapy. This clinical picture, associated with the ongoing epidemiological situation, initially guided us toward a diagnosis of COVID-19. Based on SARS-CoV-2 reverse transcription-polymerase chain reaction negativity and the absence of anti-SARS-CoV-2 antibodies, a search for a differential diagnosis was conducted that led us to conclude a diagnosis of severe pulmonary leptospirosis This case highlights the need to engage in early discussions about differential diagnoses, including neglected tropical and subtropical diseases, during the COVID-19 era.

Infectious Complications Following Snakebite by Bothrops lanceolatus in Martinique: A Case Series.

Infections secondary to snakebite occur in a number of patients and are potentially life-threatening. Bothrops lanceolatus bites in Martinique average 30 cases per year and may result in severe thrombotic and infectious complications. We aimed to investigate the infectious complications related to B. lanceolatus bite. A retrospective single-center observational study over 7 years (2011-2018) was carried out, including all patients admitted to the hospital because of B. lanceolatus bite. One hundred seventy snake-bitten patients (121 males and 49 females) were included. Thirty-nine patients (23%) presented grade 3 or 4 envenoming. Twenty patients (12%) developed wound infections. The isolated bacteria were Aeromonas hydrophila (3 cases), Morganella morganii (two cases), group A Streptococcus, and group B Streptococcus (one case each). Patients were treated empirically with third-generation cephalosporin (or amoxicillin-clavulanate), aminoglycoside, and metronidazole combinations. Outcome was favorable in all patients. The main factor significantly associated with the occurrence of infection following snakebite was the severity of envenoming (P < 0.05). Our findings clearly point toward the frequent onset of infectious complications in B. lanceolatus-bitten patients presenting with grade 3 and 4 envenoming. Thus, based on the bacteria identified in the wounds, we suggest that empiric antibiotic therapy including third-generation cephalosporin should be administered to those patients on hospital admission.

Risk Factors for Nontyphi Salmonella Bacteremia Over 10 Years in Fort-de-France, Martinique, West Indies.

Nontyphoidal Salmonella infections can result in bacteremia. This study was undertaken to determine the predictive factors for bacteremia in children aged less than 16 years. Medical data were collected for every child with positive nontyphoidal Salmonella cultures in blood or stools at the University hospital of Martinique, French West Indies, between January 2005 and December 2015. Among 454 patients, 333 were included; 156 cases had confirmed bacteremia, and 177 were included as control group with nontyphoidal Salmonella only isolated in stools. Age at diagnosis, delay before consulting, prematurity, immunosuppression, or hyperthermic seizures were not significantly associated with bacteremia. C-reactive protein was higher in cases of bacteremia (P = 0.01); however, after adjusting to the threshold of 30 mg/L, there was no longer any difference. There were also significant relations for electrolytes such as hyponatremia (odds ratio (OR) = 2.08 [95% CI = 1.31-3.95]; P < 0.01), high urea level (OR = 0.53 [95% CI = 0.32-0.88], P < 0.01). The infecting serotype was the most discriminant risk factor (P < 10-4). Among 28 serotypes isolated between 2005 and 2015, Salmonella panama was the most common serotype: 122 strains (78.2%) were isolated from bacteremic patients versus 60 (33.9%) from nonbacteremic patients (P < 10-4). Salmonella panama was the most important risk factor for bacteremia (OR = 7.37 [95% CI = 3.18-17.1], P < 10-4) even after multivariate analysis (OR = 13.09 [95% CI = 5.42-31.59], P < 10-4). After adjusting for bacteremia, S. panama was associated with a significantly higher body temperature than other Salmonella: 39°C (standard deviation [SD] = 0.92) versus 38.2°C [SD = 1.1], linear regression P < 10-3. Children with Salmonella serotype panama infection were at higher risk of bacteremia than children infected with other Salmonella serotypes.

Achromobacter xylosoxidans in hospital environments: still waters run deep!

BACKGROUND: Hospital reservoirs of Achromobacter xylosoxidans, responsible for nosocomial infections, are poorly known. METHODS: We examined the growth, survival and biofilm formation of five A. xylosoxidans strains for up to 2 y in distilled, dialysis or microfiltered water. Each strain was inoculated at 102 CFU/ml without adding nutrients. RESULTS: All strains grew at a level of 3x103 to 1.5x107 CFU/ml; each strain showed a preferred water type. Strains isolated from quaternary ammoniums showed the highest ability to grow and form biofilms in nutrient-poor waters. CONCLUSION: Medical waters and notably sterile distilled water bottles appear to be long-lasting reservoirs of A. xylosoxidans.

Achromobacter spp. healthcare associated infections in the French West Indies: a longitudinal study from 2006 to 2016.

BACKGROUND: Bacteria of the Achromobacter genus, more particularly xylosoxidans species, are responsible for various healthcare associated infections (HAI) which are increasingly described since the last decade. Cystic fibrosis (CF) patients are considered as potential reservoirs in hospitals. We performed a retrospective study to estimate the frequencies of Achromobacter spp. HAI among patients from French West Indies, to determine characteristics of infected patients and establish a possible link between CF and infections. METHODS: All adults with at least one Achromobacter spp. positive sample and infection criteria in accordance with European official definitions of HAI, hospitalized in University Hospital of Martinique from 2006 to 2016 for more than 48 h, were included. Patient clinical features, immune status and underlying diseases were obtained from medical files. A list of CF patients was given by clinicians. Antibiotic-susceptibility profiles of the strains were determined using an automated method. RESULTS: Mean incidence density was 0.038/1000 days of hospitalization. Achromobacter spp. HAI evolved as an endemic situation with a low but pretty much stable incidence rate over the 11-year observation period. An epidemic peak was noticed in 2013. Among the 66 included patients, 56.1% were immunocompetent and no one had CF. Pneumonia and bacteraemia were the two main HAI. Among the 79 isolated strains, 92.4% were resistant to at least 1 major antibiotic and 16.4% met the definition of multidrug-resistant bacteria. CONCLUSIONS: This microorganism, little known in our country because of the scarcity of CF patients, represents a threat for both immunosuppressed and immunocompetent patients and a therapeutic challenge because of its high resistance.

Achromobacter xylosoxidans resistance to antiseptics and disinfectants is far from obvious.

BACKGROUND: Achromobacter xylosoxidans is described as being resistant to antiseptics and disinfectants. We studied in vitro the ability of five strains to survive and grow in such solutions, with and without starvation. METHODS: Bacterial suspensions in rich media and in distilled water were inoculated into eight antiseptics or disinfectants under conditions of use. RESULTS: All strains from cultures in distilled water survived in aqueous chlorhexidine and only environmental strains survived in a quaternary ammonium-based disinfectant. Survival did not exceed 30 min and no growth was observed. CONCLUSIONS: This study highlights a relationship between starvation and survival in antiseptics and disinfectants.

Oral Microbiota of the Snake Bothrops lanceolatus in Martinique.

In Martinique, Bothrops lanceolatus snakebite, although relatively uncommon (~30 cases/year), may result in serious complications such as systemic thrombosis and local infections. Infections have been hypothesized to be related to bacteria present in the snake's oral cavity. In this investigation, we isolated, identified, and studied the susceptibility to beta-lactams of bacteria sampled from the oral cavity of twenty-six B. lanceolatus specimens collected from various areas in Martinique. Microbiota from B. lanceolatus oral cavity was polymicrobial. Isolated bacteria belonged to fifteen different taxa; the most frequent being Aeromonas hydrophyla (present in 50% of the samples), Morganella morganii, Klebsiella pneumoniae, Bacillus spp., and Enterococcus spp. Analysis of antibiotic susceptibility revealed that 66.7% of the isolated bacteria were resistant to amoxicillin/clavulanate. In contrast, the majority of isolated bacteria were susceptible to the third-generation cephalosporins (i.e., 73.3% with cefotaxime and 80.0% with ceftazidime). Microbiota from B. lanceolatus oral cavity is polymicrobial with bacteria mostly susceptible to third-generation cephalosporins but rarely to amoxicillin/clavulanate. In conclusion, our findings clearly support that first-line antibiotic therapy in the B. lanceolatus-bitten patients, when there is evidence of infection, should include a third-generation cephalosporin rather than amoxicillin/clavulanate.

Mediastinitis superinfected by Achromobacter xylosoxidans. A case report.

We describe an extremely rare case of mediastinitis superinfected by emerging Achromobacter xylosoxidans. After mitral and aortic valves replacement, the patient first developed a Staphylococcus aureus mediastinitis, and five days after starting adapted antibiotic therapy, superficial pus analysis revealed the presence of Achromobacter xylosoxidans. This superinfection was considered superficial and focus was made on Staphylococcus aureus mediastinitis. Three weeks later, no more Staphylococcus aureus was found in pus samples and the sepsis seemed under control. Unfortunately, blood cultures were again positive for Achromobacter xylosoxidans three weeks later and the patient died from septic shock.

Infective Endocarditis in French West Indies: A 13-Year Observational Study.

We conducted an observational study to describe the characteristics of infective endocarditis (IE) in French West Indies (FWI) and to identify variables associated with in-hospital case fatality. The records of the patients admitted for IE to the University Hospital of Martinique between 2000 and 2012 were collected using an electronic case report form. Only Duke-Li definite cases were considered for this analysis. Variables associated with in-hospital mortality were tested using univariate logistic regression analysis. The analysis included 201 patients (median age 58 years, sex ratio: two males to one female). There was no previously known heart disease in 45.8%, a prosthetic valve in 21.4%, and previously known native valve disease in 32.8% of the cases. Community-acquired IE represented 59.7% of all cases, health-care-associated IE represented 38.3% and injection-drug-use-acquired IE represented 1.5%. Locations of IE were distributed as follows: 42.3% were mitral valve IE, 34.8% were aortic valve IE, and 7% were right-sided IE. Microorganisms recovered from blood cultures included 30.4% streptococci, 28.9% staphylococci, and 5% enterococci. Blood cultures were negative in 20.9% of the cases. Surgical treatment was performed in 53% of the patients. In-hospital case fatality rate was 19%. Advanced age, Staphylococcus aureus IE, and health-care-associated IE were associated with in-hospital case fatality. The epidemiological and microbiological profile of IE in FWI is in between those observed in developed countries and developing countries: patients were younger, blood cultures were more frequently negative, and IE due to group D streptococci and enterococci were less common than in industrialized countries.

Evaluation of different serological assays for early diagnosis of leptospirosis in Martinique (French West Indies).

BACKGROUND: Leptospirosis is a potentially life-threatening but curable zoonosis whose prognosis depends on accurate and timely diagnosis. Because of its non-specific clinical presentation, laboratory testing is essential to confirm the diagnosis. Here, we aimed to assess the performance of two enzyme-linked immunosorbent assays (ELISAs) (ELISA Serion and ELISA-Hb Pasteur) and one immunodot (GenBio) using quantitative PCR (qPCR) as gold standard, instead of the traditional microscopic agglutination test, for the diagnosis of acute leptospirosis in an endemic area. METHODOLOGY/PRINCIPAL FINDINGS: Between January 2011 and December 2012, a total of 122 patients were diagnosed with leptospirosis, as confirmed by qPCR at the University Hospital of Martinique. Among them, 103 had at least one serum sample available for analysis. Performance of each serological assay was evaluated according to days' post onset of symptoms (DPO) and local species diversity (which included L. santarosai, L. interrogans, L. kirschneri, L. borgpetersenii, L. noguchii, and L. kmetyi). Several thresholds were tested to optimize accuracy. When considering the manufacturer's threshold, the sensitivity of ELISA Serion, ELISA-Hb Pasteur and GenBio immunodot was 75%, 67% and 64%, while specificity was 92%, 98% and 100%, respectively. Moreover, the threshold optimization allowed a significant improvement in specificity for the ELISA Serion from 92% to 99% (p<0.05). During the first 5 DPO, sensitivities were 35%, 30% and 42% for ELISA Serion, ELISA-Hb Pasteur and GenBio immunodot, respectively. However, between 6─10 DPO, these sensitivities dramatically increased to reach 86%, 76% and 67%, respectively. Performances of the three assays were not affected by the species studied. CONCLUSIONS/SIGNIFICANCE: All these serological assays showed the potential for diagnosing leptospirosis after (but not before) 6 days' post onset of symptoms. In a high prevalence setting, where highest specificities are needed, threshold optimizing should be performed for this purpose.

Factors Associated with Severe Leptospirosis, Martinique, 2010-2013.

To identify factors associated with disease severity, we examined 102 patients with quantitative PCR-confirmed leptospirosis in Martinique during 2010-2013. Associated factors were hypotension, chest auscultation abnormalities, icterus, oligo/anuria, thrombocytopenia, prothrombin time <68%, high levels of leptospiremia, and infection with L. interrogans serovar Icterohaemorrhagiae/Copenhageni.

Major West Indies MRSA clones in human beings: do they travel with their hosts?

BACKGROUND: Descriptions of the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) have seldom been produced in the Caribbean, which is a major tourism destination. MATERIALS AND METHODS: Using DNA microarrays and spa typing, we characterized 85 MRSA isolates from human skin and soft-tissue infections from five different islands. RESULTS: In the French West Indies (n = 72), the most frequently isolated clones were the same clones that are specifically isolated from mainland France [Lyon (n = 35) and Geraldine (n = 11) clones], whereas the clones that were most frequently isolated from the other islands (n = 13) corresponded with clones that have a worldwide endemic spread [Vienna/Hungarian/Brazilian (n = 5), Panton Valentine leukocidin-positive USA300 (n = 4), New York/Japan (n = 2), and pediatric (n = 1) clones]. CONCLUSION: The distribution of the major MRSA clones in the French (Guadeloupe and Martinique) and non-French West Indies (Jamaica, Trinidad, and Tobago) is different, and the clones most closely resemble those found in the home countries of the travelers who visit the islands most frequently. The distribution might be affected by tourist migration, which is specific to each island.

Human infection with Shewanella putrefaciens and S. algae: report of 16 cases in Martinique and review of the literature.

Shewanella spp. are saprophytic bacteria that are part of the marine microflora in warm climates and are rarely pathogenic. However, Shewanella spp. infections are being increasingly reported, and there has been no comprehensive review of the literature describing these infections. This article reports 16 cases of Shewanella spp. infections in Martinique since 1997 and reviews another 239 cases reported in the literature since 1973. Patients experienced soft tissue infections, ear infection, or abdominal and biliary tract infections. A skin or mucosal portal of entry was found for 53% of the patients and exposure to the marine environment was reported for 44%; 79% of patients had an underlying condition. Bacteriema were frequent (28%). Most (87%) patients recovered, although ear infections can become chronic. Death occurred in 13% of the patients. Most Shewanella spp. isolates are susceptible to cefotaxime (95%), piperacillin and tazobactam (98%), gentamicin (99%), and ciprofloxacin (94%).

Serovar diversity of pathogenic Leptospira circulating in the French West Indies.

BACKGROUND: Leptospirosis is one of the most important neglected tropical bacterial diseases in Latin America and the Caribbean. However, very little is known about the circulating etiological agents of leptospirosis in this region. In this study, we describe the serological and molecular features of leptospires isolated from 104 leptospirosis patients in Guadeloupe (n = 85) and Martinique (n = 19) and six rats captured in Guadeloupe, between 2004 and 2012. METHODS AND FINDINGS: Strains were studied by serogrouping, PFGE, MLVA, and sequencing 16SrRNA and secY. DNA extracts from blood samples collected from 36 patients in Martinique were also used for molecular typing of leptospires via PCR. Phylogenetic analyses revealed thirteen different genotypes clustered into five main clades that corresponded to the species: L. interrogans, L. kirschneri, L. borgpetersenii, L. noguchi, and L. santarosai. We also identified L. kmetyi in at least two patients with acute leptospirosis. This is the first time, to our knowledge, that this species has been identified in humans. The most prevalent genotypes were associated with L. interrogans serovars Icterohaemorrhagiae and Copenhageni, L. kirschneri serovar Bogvere, and L. borgpetersenii serovar Arborea. We were unable to identify nine strains at the serovar level and comparison of genotyping results to the MLST database revealed new secY alleles. CONCLUSIONS: The overall serovar distribution in the French West Indies was unique compared to the neighboring islands. Typing of leptospiral isolates also suggested the existence of previously undescribed serovars.

Clinical and microbiologic characteristics of children treated at the Fort de France university hospital after the 2010 Haiti earthquake.

Few data related to the care of children injured in an earthquake are available. The objective of our study was to analyze the results of clinical and microbiologic characteristics of children treated in a Fort de France hospital after the Haiti earthquake. Bacteria were mainly Gram-negative bacteria. Some of these infections seemed to be environmental; a significant portion was related to acquisition during hospitalization.

Bacteremia caused by Aeromonas species [corrected] complex in the Caribbean Islands of Martinique and Guadeloupe.

Aeromonas species are Gram-negative bacilli of the water environment whose survival appears facilitated by warm climates. There have been no reports on Aeromonas species in the [corrected] Caribbean to date. Our aim was to describe clinical and bacteriological features in patients presenting with such bacteremia in Martinique and Guadeloupe. During a 14-year period, we retrospectively identified 37 patients. The mean age was 55 years and in 89% of cases underlying disease such as digestive diseases, cutaneous wounds, and malignancy were identified. One case was related to severe strongyloidiasis and one with snake bite. Polymicrobial bacteremia was identified in 38%, essentially with Enterobacteriaceae. All Aeromonas isolates were resistant to amoxicillin but extended-spectrum beta-lactam and fluoroquinolone were active against more than 95%. During hospitalization 10 patients died (27%). Older age, occurrence of multiorgan failure, and impaired renal function were associated with in-hospital mortality.