First isolation of Leptospira noguchii serogroups Panama and Autumnalis from cattle.

Prevention and control of leptospirosis are based on the knowledge of locally circulating strains. Thus, efforts to obtain local isolates are paramount to the epidemiological understanding of leptospirosis. We report and discuss here the first isolation of members of serogroups Autumnalis and Panama from cattle, both belonging to Leptospira noguchii species. Urine samples (n = 167) were collected directly by puncture of the bladder from randomly selected cows from a slaughterhouse in Rio de Janeiro, Brazil, for bacteriological culture. Isolates were characterized by serogrouping and sequencing (rrs and secY genes). Overall, 10/167 positive urine samples (6%) were obtained. Sequencing of amplicons targeting for both rrs and secY genes identified two of them (2013_U73 and 2013_U232) as L. noguchii. Serogrouping of those strains indicated that 2013_U73 belonged to the Panama serogroup (titre 1600), and 2013_U232 to the Autumnalis serogroup (titre 12800). Both Panama and Autumnalis are known agents of incidental leptospirosis in cattle. This group of leptospires could be particularly important in tropical countries. This is the first report of members of serogroups Autumnalis and Panama belonging to L. noguchii species from cattle. Although related to previously reported strains, these isolates have been shown to be genetically diverse from them.

Antimicrobial resistance of Listeria monocytogenes strains isolated from humans in France.

Susceptibility to antibiotics of 4,816 clinical L. monocytogenes strains isolated since 1926 was studied, and the temporal evolution of susceptibility to antibiotics was analyzed through several decades. The mechanisms of resistance in each resistant strain were studied. The prevalence of resistant strains was estimated at 1.27% among isolates from humans. Resistance to tetracyclines+ and fluoroquinolones was more common and has recently emerged. Although acquired resistance in clinical L. monocytogenes did not implicate clinically relevant antibiotics, the possibility of resistance gene transfers, the description of the first clinical isolate with high-level resistance to trimethoprim, and the recent increase in penicillin MICs up to 2 microg/ml reinforce the need for microbiological surveillance.

Real-time PCR detection of gyrA and parC mutations in Streptococcus pneumoniae.

Fluoroquinolone resistance in Streptococcus pneumoniae mainly involves stepwise mutations predominantly in the parC and gyrA genes. We have developed a single-run real-time PCR assay for detection of the four most common mutations in the quinolone resistance-determining regions of these genes. This assay provides a useful tool for both clinical and epidemiological use.

First isolation of Yersinia entomophaga in human urinary tract.

Yersinia entomophaga is an insect pathogen first isolated from larvae of Coleoptera in New Zealand in 2011. We report here the first isolation of Y. entomophaga from human urine. Using whole-genome sequencing, we confirmed the presence of specific chromosomal virulence genes and identified a plasmid harbouring a quinolone resistance gene.

Molecular analysis of leptospires from serogroup Sejroe obtained from asymptomatic cattle in Rio de Janeiro - Brazil reveals genetic proximity to serovar Guaricura.

Bovine leptospirosis causes substantial reproductive failure in cattle, mainly due to infections with serovar (sv) Hardjo infection. Notwithstanding, other serovars from the serogroup (sg) Sejroe could also have important roles in bovine leptospirosis. The objective was to investigate genetic diversity of serogroup Sejroe isolates obtained from asymptomatic cattle in the state of Rio de Janeiro, Brazil. Urine and vaginal fluid (VF) were collected from clinically healthy cattle immediately after slaughter. Five isolates were recovered and characterized (serogrouping) as belonging to sg Sejroe. Sequencing of rrs and secY genes further identified them as Leptospira santarosai. Analysis of secY sequences indicated a high level of sequence homology to sv Guaricura strains. Based on culture and sequence data, we inferred that other members of sg Sejroe may be important in bovine leptospiral infection, particularly genotypes of L. santarosai serovar Guaricura.

Molecular Characterization and Serology of Leptospira kirschneri (Serogroup Grippotyphosa) Isolated from Urine of a Mare Post-Abortion in Brazil.

A strain of Leptospira kirschneri (serogroup Grippotyphosa) was cultured from urine of a mare post-abortion in Brazil and characterized by serogrouping, multiple-locus variable-number tandem repeat analysis, PGFE, and sequencing of genes rrs and secY. Strains of L. kirschneri have apparently never been recovered from horses in tropical area, only in Europe and USA. Knowledge of local epidemiology is important to interpret genetic profiles of leptospires circulating in an area.

Detection, seroprevalence and antimicrobial resistance of Yersinia enterocolitica and Yersinia pseudotuberculosis in pig tonsils in Northern Italy.

Yersiniosis is the third most common reported zoonoses in Europe, with Y. enterocolitica and Y. pseudotuberculosis responsible for 98.66% and 0.94% of the confirmed human cases in 2013. From June 2013 to October 2014, 201 pigs at slaughter belonging to 67 batches were tested for Y. enterocolitica and Y. pseudotuberculosis in tonsils. Diaphragm muscle samples were tested for antibodies against Yersinia by a commercially available ELISA test. Y. enterocolitica 4/O:3 was detected in 55/201 pig tonsils (27.4%; 95% CI 23.1-37.1). The positive pigs came from 38/67 batches (56.7%) and were reared in 36/61 farms (59.0%). There was no statistical difference between farrow-to-finish and finishing farms. The mean count of Y. enterocolitica was 3.56±0.85log10CFU/g with a minimum of 2.0log10CFU/g and a maximum of 4.78log10CFU/g. Y. pseudotuberculosis was isolated from 4/201 pig tonsils (2.0%; 95% CI 0.0-4.5). Three isolates belonged to serotype O:3 and one to serotype O:1. The positive pigs belonged to 4/67 batches (6.0%) and came from finishing farms only. Y. pseudotuberculosis could be enumerated in one sample only (4.27log10CFU/g). The ELISA test demonstrated that 56.1% of the meat juice samples were positive for Yersinia antibodies. Serological positivity was found in 67.9% (36/53) of the Y. enterocolitica- and 75.0% (3/4) of the Y. pseudotuberculosis positive pigs. A significant association was found between serological results and the presence of Y. enterocolitica in tonsils (OR=1.97, p=0.044). All the Y. enterocolitica 4/O:3 isolates were susceptible to amoxicillin-clavulanic acid, gentamicin, ceftazidime, ertapenem and meropenem, 94.5% to cefotaxime, 89.1% to kanamycin and 78.2% to tetracycline. The highest resistance rates were observed for ampicillin (100%), sulphonamides (98.2%) and streptomycin (78.2%). Y. pseudotuberculosis strains were sensitive to all the antimicrobials tested, i.e. amoxicillin, amoxicillin/clavulanic acid, azithromycin, cephalothin, cefoxitin, ceftriaxone, ciprofloxacin, nalidixic acid, sulphonamide, tetracycline and ticarcillin. The study shows that Italian fattening pigs are frequently infected with human pathogenic Y. enterocolitica 4/O:3. Although the isolation rate is slightly lower than in other European countries, the serological test demonstrates that the infection is widespread among pig population. In fact, seroprevalence is similar to other EU countries. The detection of Y. pseudotuberculosis serotypes O:1 and O:3 in pig tonsils is of concern. Since tonsils may represent a contamination source for pig meat at slaughter, further studies regarding human infections by both microbial species are strongly recommended.

In vitro antibacterial activity of ceftobiprole against clinical isolates from French teaching hospitals: proposition of zone diameter breakpoints.

The aims of this study were to determine the in vitro activity profile of ceftobiprole, a pyrrolidinone cephalosporin, against a large number of bacterial pathogens and to propose zone diameter breakpoints for clinical categorisation according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) minimum inhibitory concentration (MIC) breakpoints. MICs of ceftobiprole were determined by broth microdilution against 1548 clinical isolates collected in eight French hospitals. Disk diffusion testing was performed using 30 µg disks according to the method of the Comité de l'Antibiogramme de la Société Française de Microbiologie (CA-SFM). The in vitro activity of ceftobiprole, expressed by MIC(50/90) (MICs for 50% and 90% of the organisms, respectively) (mg/L), was as follows: meticillin-susceptible Staphylococcus aureus, 0.25/0.5; meticillin-resistant S. aureus (MRSA), 1/2; meticillin-susceptible coagulase-negative staphylococci (CoNS), 0.12/0.5; meticillin-resistant CoNS, 1/2; penicillin-susceptible Streptococcus pneumoniae, ≤ 0.008/0.03; penicillin-resistant S. pneumoniae, 0.12/0.5; viridans group streptococci, 0.03/0.12; ß-haemolytic streptococci, ≤ 0.008/0.016; Enterococcus faecalis, 0.25/1; Enterococcus faecium, 64/128; Enterobacteriaceae, 0.06/32; Pseudomonas aeruginosa, 4/16; Acinetobacter baumannii, 0.5/64; Haemophilus influenzae, 0.03/0.12; and Moraxella catarrhalis, 0.25/0.5. According to the regression curve, zone diameter breakpoints could be 28, 26, 24 and 22 mm for MICs of 0.5, 1, 2 and 4 mg/L respectively. In conclusion, this study confirms the potent in vitro activity of ceftobiprole against many Gram-positive bacteria, including MRSA but not E. faecium, whilst maintaining a Gram-negative spectrum similar to the advanced-generation cephalosporins such as cefepime. Thus ceftobiprole appears to be well suited for the empirical treatment of a variety of healthcare-associated infections.

Klebsiella pneumoniae LO10 producing extended-spectrum beta-lactamase SHV-12 in Saudi Arabia.

Klebsiella pneumoniae LO10 was responsible for an outbreak that occurred in the neonatal unit at Security Forces Hospital, Kingdom of Saudi Arabia. Over a period of eight months nine cases of bacteremia resulted in two deaths. Resistance to third generation cephalosporins was transferred from strain LO10 to E. coli by both conjugation and transformation. Sequence determination of the plasmid gene from two transconjugants and one transformant indicated that resistance was carried by a ca.100-kb plasmid encoding beta-lactamase SHV-12. This is the first description of a K. pneumoniae producing a type SHV-12 extended spectrum beta-lactamase in Riyadh. Long term exposure to antibiotics, prolonged stay, and heavy use of third generation cephalosporins contributed to the spread of the resistant strain in the unit. Strict infection control measures led to control of the outbreak.

[Qnr-type quinolone resistance in extended-spectrum beta-lactamase producing enterobacteria in Abidjan, Ivory Coast]. / Résistance aux quinolones de type qnr chez les entérobactéries productrices de bêta-lactamases à spectre élargi à Abidjan en Côte d'Ivoire.

The aim of the study was to show the emergence of the qnr genes in extended spectrum beta-lactamases producing enterobacteria in Abidjan between 2005 and 2006. The whole of 151 strains of extended spectrum beta-lactamases producing enterobacteria were studied: 64 Escherichia coli, 66 Klebsiella pneumoniae, seven Klebsiella oxytoca and 14 Enterobacter spp. isolated from various biological products and from in- and out-patients. The techniques of disks diffusion, double-disk synergy, E-test were respectively used for the antimicrobial susceptibility test, the detection of extended spectrum beta-lactamases and the minimal inhibiting concentration. The bla genes(SHV, TEM, CTXM groups 1, 2, 8, 9), and AmpC were determined by PCR and characterized by sequencing. A global prevalence of 27,2 % (41/151) and rates of 9,9, 14,6, 2,7 % for the qnr genes A, B, A and S were observed. The distribution was 42,9 % for Enterobacter spp, 31,2 % for Escherichia coli, 20,5 % for Klebsiella; 30 strains expressed at least two bla genes; four strains were associated with AmpC. The strains were resistant to the cotrimoxazole (97,6 %), to the céfépime (73,2 %), to the céfoxitine (56,1 %), to the imipénème (0 %) and 43,9 % to all the aminosides. This high qnr gene prevalence associated with several types of bla genes in epidemic matter, the high level of resistance to antibiotics make fear a high risk of the transmission of multi-resistants bacteria and challenge the authorities for a resistance monitoring policy.