National survey of colistin resistance among carbapenemase-producing Enterobacteriaceae and outbreak caused by colistin-resistant OXA-48-producing Klebsiella pneumoniae, France, 2014

From January 2014 to December 2014, 972 consecutive non-replicate carbapenemase-producing Enterobacteriaceae isolates from colonised or infected patients were collected at the Associated French National Reference Centre as part of the French national survey on antimicrobial resistance. It included 577 Klebsiella spp. (59%), 236 Escherichia coli (24%), 108 Enterobacter spp. (11%), 50 Citrobacter spp. (5%), and a single Salmonella spp. isolate (0.1%). Of 561 K. pneumoniae isolates, 35 were found to be resistant to colistin (6.2%). PFGE analysis revealed a clonal outbreak involving 15 K. pneumoniae isolates belonging to sequence type ST11, recovered in a single hospital in the Picardie region in northern France. Those clonally related isolates showed variable levels of resistance to colistin, ranging from 4 to 64 mg/L. They harboured the blaOXA-48 carbapenemase gene and the blaCTX-M-15 extended-spectrum beta-lactamase gene. Among the 91 Enterobacter cloacae isolates, seven were resistant to colistin and produced different types of carbapenemases. Surprisingly, none of the E. coli and Citrobacter spp. isolates showed resistance to colistin. This national survey including carbapenemase-producing isolates recovered in 2014 reported a high rate of colistin resistance in K. pneumoniae and E. cloacae (6.2% and 7.7%, respectively) in France

Antimicrobial Susceptibility Trends Observed in Urinary Pathogens Obtained From New York State

International guidelines recommend using local susceptibility data to direct empiric therapy for acute uncomplicated cystitis. We evaluated outpatient urinary isolate susceptibility trends in New York State. Nitrofurantoin had the lowest resistance prevalence whereas trimethoprim-sulfamethoxazole and fluoroquinolones had higher prevalences. This study highlights the need for local outpatient antimicrobial stewardship programs

Differences in microbiota between two multilocus lineages of the sugarcane Aphid (Melanaphis sacchari) in the continental United States

The sugarcane aphid (SCA), Melanaphis Sacchari (Zehntner) (Hemiptera: Aphididae), has been considered an invasive pest of sugarcane in the continental United States since 1977. Then, in 2013, SCA abruptly became a serious pest of U.S. sorghum and is now a sorghum pest in 22 states across the continental United States. Changes in insect-associated microbial community composition are known to influence host-plant range in aphids. In this study, we assessed whether changes in microbiota composition may explain the SCA outbreak in U.S. sorghum. We characterized the SCA bacterial microbiota on sugarcane and grain sorghum in four U.S. states, using a metabarcoding approach. In addition, we used taxon-specific polymerase chain reaction (PCR) primers to screen for bacteria commonly reported in aphid species. As anticipated, all SCA harbored the primary aphid endosymbiont Buchnera aphidicola, an obligate mutualistic bacterial symbiont. Interestingly, none of the secondary symbionts, facultative bacteria typically associated with aphids (e.g., Arsenophonus, Hamiltonella, Regiella) were present in either the metabarcoding data or PCR screens (with the exception of Rickettsiella and Serratia, which were detected by metabarcoding at low abundances <1%). However, our metabarcoding detected bacteria not previously identified in aphids (Arcobacter, Bifidobacterium, Citrobacter). Lastly, we found microbial host-associated differentiation in aphids that seems to correspond to genetically distinct aphid lineages that prefer to feed on grain sorghum (MLL-F) versus sugarcane (MLL-D)

Superficieibacter electus gen. nov., sp. nov., an extended-spectrum β-lactamase possessing member of the enterobacteriaceae family, isolated from Intensive Care Unit surfaces

<p>Two Gram-negative bacilli strains, designated BP-1(T) and BP-2, were recovered from two different Intensive Care Unit surfaces during a longitudinal survey in Pakistan. Both strains were unidentified using the bioMerieux VITEK MS IVD v2.3.3 and Bruker BioTyper MALDI-TOF mass spectrometry platforms. To more precisely determine the taxonomic identity of BP-1(T) and BP-2, we employed a biochemical and phylogenomic approach. The 16S rRNA gene sequence of strain BP-1(T) had the highest identity to Citrobacter farmeri CDC 2991-81(T) (98.63%) Citrobacter amalonaticus CECT 863(T) (98.56%), Citrobacter sedlakii NBRC 105722(T) (97.74%) and Citrobacter rodentium NBRC 105723(T) (97.74%). The biochemical utilization scheme of BP-1(T) using the Analytic Profile Index for Enterobacteriaceae (API20E) indicated its enzymatic functions are unique within the Enterobacteriaceae but most closely resemble Kluyvera spp., Enterobacter cloacae and Citrobacter koseri/farmeri. Phylogenomic analysis of the shared genes between BP-1(T), BP-2 and type strains from Kluyvera, Citrobacter, Escherichia, Salmonella, Kosakonia, Siccibacter and Shigella indicate that BP-1(T) and BP-2 isolates form a distinct branch from these genera. Average Nucleotide Identity analysis indicates that BP-1(T) and BP-2 are the same species. The biochemical and phylogenomic analysis indicate strains BP-1(T) and BP-2 represent a novel species from a new genus within the Enterobacteriaceae family, for which the name Superficieibacter electus gen. nov., sp. nov., is proposed. The type strain is BP-1(T) (= ATCC BAA-2937, = NBRC 113412).</p

Bayesian estimation of diagnostic accuracy of fecal culture and PCR-based tests for the detection of Salmonella enterica in California cull dairy cattle.

Epidemiological studies of low prevalence disease problems are often hindered by the high cost of diagnostic testing. The objective of this study was to evaluate PCR screening of both individual and pooled fecal samples from culled dairy cows for the invA gene of Salmonella followed by culture to determine if the sensitivity and specificity were comparable to the results from traditional culture methods applied to individual samples. Cows from six different dairies were sampled in all four seasons. A total of 240 individual cow fecal samples, 24 fecal pools and 24 pools of 24-hour tetrathionate enrichment broth were tested. Diagnostic sensitivity of PCR screening followed by culture of PCR positive or indeterminate samples (i.e PCR-CUL method) was lower than that of culture (CUL) when applied to individual fecal samples (94.8%, 99.5%), however the specificity was comparable (99.6% and 97.7% respectively). For pools of five fecal samples and pools of five, 24 h tetrathionate broth samples, the specificity of both tests were comparable (∼98%); however, their sensitivity was only comparable in pooled fecal samples (∼93%) but greater for culture compared to PCR-CUL in pooled broth samples (∼99% versus ∼93%). Compared to culture results from testing of individual fecal samples, testing pooled fecal samples by culture had a relative sensitivity of 74% and relative specificity of 96%, testing pooled fecal samples by PCR-CUL resulted in relative sensitivity of 90% and relative specificity of 96%. Testing of pooled 24-hour enrichment broth by PCR-CUL increased the relative sensitivity and specificity to 100%. PCR testing followed by culture of positive or indeterminate samples is a time saving alternative to traditional methods. In addition, pooling of samples may be a useful method for decreasing cost if study aims can accommodate a moderate loss of relative sensitivity

Prevalence of Gram-negative Pathogens and their antimicrobial susceptibility in bacterial meningitis in pediatric cases

The present study was conducted to find out the prevalence and spectrum of Gram negative pathogens causing bacterial meningitis and their antimicrobial susceptibility pattern in a tertiary care hospital. The cerebrospinal fluid (CSF) (3-5 ml) was collected from 638 admitted children clinically suspected of septic meningitis. Bacterial isolates were identified and antimicrobial susceptibility was assessed by the Kirby-Bauer disk diffusion method. Of the 638 samples tested 102 (15.99%) were culture positive. Male to female (M:F) ratio was 1.62:1. The maximum incidence of 45 (44.12%) cases was found in children (1-12 yrs); in institutional deliveries the incidence was 58 (56.86%) cases. Further, the incidence of 51 cases was found from May to August. Escherichia coli (E. coli) were commonest, seen in 9 (25%) cases followed by Acinetobacter spp., Citrobacter spp. and Klebsiella spp. with 6 (16.67%) cases each. Enterobacter spp., Neisseria spp. and Pseudomonas aeruginosa were isolated in 3 (8.33%) cases each. E. coli, Acinetobacter spp, Citrobacter spp and Klebsiella spp isolates were 100% susceptible to meropenem, piperacillin-tazobactam and cefoperazone-sulbactam and 100% resistant to cotrimoxazole and tetracycline. All strains of Neisseria spp, Enterobacter spp and Pseudomonas spp. were 100% susceptible to meropenem followed by gatifloxacin. These were 100% resistant to tetracycline and cotrimoxazole. Neisseria spp. were also 100% susceptible to pristinamycin. In septic meningitis Gram negative organisms are less common (35.29%). Of the isolates, more common Gram negative isolates included E. coli, Acinetobacter Spp., Citrobacter Spp., and Klebsiella spp. and these isolates were 100% susceptible to meropenem, piperacillin-tazobacatam and cefoperazone-sulbactam. Hence, empirical therapy should be formulated according to antimicrobial susceptibility patterns

Broad-spectrum β-lactamases among Enterobacteriaceae of animal origin: molecular aspects, mobility and impact on public health

Broad-spectrum β-lactamase genes (coding for extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases) have been frequently demonstrated in the microbiota of food-producing animals. This may pose a human health hazard since these genes may be present in zoonotic bacteria, which would cause a direct problem. They can also be present in commensals, which may act as a reservoir of resistance genes for pathogens causing disease both in humans and animals. Broad-spectrum β-lactamase genes are frequently located on mobile genetic elements, such as plasmids, transposons and integrons, which often also carry additional resistance genes. This could limit treatment options for infections caused by broad-spectrum β-lactam-resistant microorganisms. This review addresses the growing burden of broad-spectrum β-lactam resistance among Enterobacteriaceae isolated from food, companion and wild animals worldwide. To explore the human health hazard, the diversity of broad-spectrum β-lactamases among Enterobacteriaceae derived from animals is compared with respect to their presence in human bacteria. Furthermore, the possibilities of the exchange of genes encoding broad-spectrum β-lactamases – including the exchange of the transposons and plasmids that serve as vehicles for these genes – between different ecosystems (human and animal) are discussed

Sepsis caused by bloodstream infection in patients in the intensive care unit: the impact of inactive empiric antimicrobial therapy on outcome

Background: Sepsis is one of the leading causes of death in the UK. Aims: The aims of this study were to identify the rate of inactive antimicrobial therapy (AMT) in the ICU and whether inactive AMT had an effect on in hospital mortality, ICU mortality, 90-day mortality and length of hospital stay. Additionally, we wanted to identify risk factors for receiving inactive AMT. Methods: This was a retrospective observational study conducted at Glasgow Royal Infirmary ICU between January 2010 and December 2013, with 12,000 blood cultures taken over this time period, of which n=127 were deemed clinically significant. Multivariate logistic regression was used to identify risk factors independently associated with mortality. To identify risk factors for receiving inactive AMT a univariable and a subsequent multivariate analysis was constructed. Results: The rate of inactive AMT was 47% (n =60). Our multivariate analysis showed that receiving antibiotics within the first 24 hours of ICU admission led to a reduced mortality (RR 1.70; 95% CI 1.19-2.44.) Furthermore, it showed that severity of illness (as defined by SIRS criteria sepsis vs septic shock) increased mortality (OR 9.87; 95% CI 1.73-55.5). However, inactive AMT did not increase mortality (OR 1.07; 95% CI 0.47-2.41) or length of hospital stay (53.2 vs 69.1 days p=0.348.) We identified fungal bloodstream infection as a risk factor for receiving inactive AMT (OR 5.10;95% CI 1.29-20.14. Conclusion: Mortality from sepsis is influenced by multiple factors. We were unable to demonstrates that inactive AMT had an effect on mortality in sepsis

Unique features of Plasmids among different Citrobacter species

The _Citrobacter_ plasmids are supposed to represent the host genetic association within the living bacterial cell. The plasmids impart various beneficial characteristics to the host, helping it to retain suitable characteristics for adaptation as well as evolution. The study aims at understanding the role of prophage in influencing host functional characteristics by horizontal gene transfer or as whole plasmids. The _Citrobacter_ plasmid can be understood by analyzing many hypothetical protein sequences within its genome. Our study included 82 hypothetical proteins in 5 _Citrobacter_ plasmids genomes. The function predictions in 31 hypothetical proteins and 3-D structures were predicted for 11 protein sequences using PS2 server. The probable function prediction was done by using Bioinformatics web tools like CDD-BLAST, INTERPROSCAN, PFAM and COGs by searching sequence databases for the presence of orthologous enzymatic conserved domains in the hypothetical sequences. This study identified many uncharacterized proteins, whose roles are yet to be discovered in _Citrobacter_ plasmids. These results for unknown proteins within plasmids can be used in linking the genetic interactions of _Citrobacter_ species and their functions in different environmental conditions