First report of New Delhi metallo-ß-lactamase-1-producing strains in Slovakia.

Occurrence of carbapenemase-producing organisms, including New Delhi metallo-ß-lactamase-1 (NDM-1) is increasingly reported worldwide. The aim of this study was to assess the distribution of carbapenemase producers among multidrug-resistant Gram-negative bacteria isolated from blood cultures. All carbapenem-resistant strains collected from December 2011 to December 2012 were analyzed. Presence of carbapenemases was assessed with combined disc test and Carba NP test followed by polymerase chain reaction for carbapenemase genes. Altogether, 30 strains were detected, of which 7 were positive for VIM (23.3%), 6 for NDM-1 (20%), 5 for IMP (16.7%), and KPC was present in one isolate (3.3%). Four Pseudomonas aeruginosa strains were found to produce more than one carbapenemase. We also present the case report of a patient with Acinetobacter baumannii ventilator-associated pneumonia, followed by sepsis due to Enterococcus faecalis and pan-resistant NDM-1-producing P. aeruginosa. Despite the inappropriate therapy, the patient was successfully treated. This is the first report of NDM-1-producing strains in Slovakia and it contributes to a number of studies mapping the distribution of carbapenemase producers in Europe.

[New antibiotics: do we need new molecules or new strategies?]. / Nové antibiotiká: potrebujeme nové molekuly, alebo nové stratégie?

UNLABELLED: Even though the situation in the field of research and development of new antimicrobial agents is not ideal, the years of stagnation, especially among anti-gramnegative agents, seem to be over. During the years 2011-2012 we have witnessed a movement towards a development of some new antibiotic agents not only with anti-gramposi-tive activity but with anti-gramnegative too, as well as new antituberculotics and antifungals. Here we present some of these new antiinfectives, presented in conferences during the last year, of which many are being shifted to the level of clinical trials of phase II-III, while others are in stage of preclinical studies. KEYWORDS: antibiotics - antimycotics - antituberculotics.

Transferable fluoroquinolone resistance in Enterobacteriaceae and Pseudomonas aeruginosa isolated from hemocultures.

BACKGROUND: The main mechanisms causing high-level resistance to fluoroquinolones (FQ) are encoded chromosomally; that includes mutations in genes coding DNA-gyrase, but overexpression of efflux pumps contributes to increased minimum inhibitory concentration (MIC) of FQ as well. However, genes responsible for FQ-resistance may be harboured in transferable/conjugative plasmids. For some time, there was an assumption that resistance to FQ cannot be transferable in conjugation due to their synthetic origin, until 1998, when plasmid-mediated resistance transmission in Klebsiella pneumoniae was proved. We aimed to detect the occurrence of transferable FQ-resistance among Gram- negative bacteria isolated from patients in Czech and Slovak hospitals. METHODS: In this study, we tested 236 clinical isolates of Gram-negative bacteria for transferable resistance. Among relevant isolates we performed PCR detection of transferable fluoroquinolone genes (qnr). RESULTS: We have observed transfer of determinants of cephalosporin-resistance, aminoglycoside resistance as well as FQ-resistance (in 10 cases; 4.24%) not only intra-species but inter-species too. The presence of qnr gene was detected in two isolates of forty tested (5%). We have also observed that determinants of cephalosporin-resistance and aminoglycoside-resistance were linked to those of FQ-resistance and were transferred en block in conjugation. CONCLUSION: We have proved that resistance to fluoroquinolones can be transferred horizontally via conjugation among Gram-negative bacteria of different species and is associated with resistance to other antibiotics.

Nasopharyngeal bacterial colonisation in HIV-positive children in Cambodia.

OBJECTIVE: To determine patterns of nasopharyngeal colonisation in HIV-positive children. METHODS: Nasopharyngeal, nasal and ear swabs were prospectively taken from all children living in two paediatric nursing homes for HIV-positive orphans in Cambodia from 2004 to 2011. RESULTS: A total of 882 swabs were taken, of which 586 tested positive for bacteria. Staphylococcus aureus was the most frequently isolated species (178 isolates; 30.4%) followed by Streptococcus pneumoniae (103 isolates; 17.6%) and Klebsiella pneumoniae (99 isolates; 16.9%). The rate of S. pneumoniae decreased in 2009 when a vaccination programme was introduced. CONCLUSIONS: The respiratory tract of HIV-positive children receiving highly active antiretroviral therapy is commonly colonised by S. aureus and S. pneumoniae, while other species normally found in the respiratory tract, such as Moraxella catarrhalis, are far less frequent.

Tropical neuroinfections in south sudanese rural hospitals - analysis of 8,709 patients.

OBJECTIVE: Infections involving the central nervous system have very serious consequences and affect thousands of people in Africa. Despite the availability of new antibiotics and vaccines, neuroinfections act as dangerous and life-threatening conditions. The most frequent neuroinfections which are of the greatest importance for public health systems are viral diseases (such as HIV, encephalitis, poliomyelitis, rabies), bacterial diseases (bacterial meningitis, neurological complications of leprosy and tuberculosis) and parasitic infections (cerebral malaria, sleeping sickness, trypanosomiasis, schistosomiasis, toxoplasmosis etc.). METHODS: A descriptive study to assess the occurrence of neuroinfections in two rural hospitals in Sudan (Mapuordit in Yirol and Gordim in Aweil) was performed in two periods of two years: (i) 2005-2006 and (ii) 2010-2011. We obtained data on patients from Mapuordit and from Gordim by studying their medical records. RESULTS: Several cases of neuroinfections were observed during both periods; those were represented by tetanus, meningococcal meningitis, leprosy with neuropathy (altogether 442 patients) in Mapuordit. Also in Gordim, severe neuroinfections such as cerebral malaria were very rare (1 case), as well as tetanus (1 case), meningococcal meningitis (8 cases) and sleeping sickness (9 cases). However, the incidence of neuroinfections decreased from 44/1000 in 2005-2006 to 2/1000 in 2010-2011. CONCLUSIONS: Decreased incidence of serious neuroinfections (cerebral malaria, sleeping sickness, meningococcal meningitis) in Sudan may be related to improvement of effective therapeutic options, represented by (i) intermittent preventive therapy (IPT) for malaria, (ii) by suppression of sleeping sickness vectors and (iii) by better accessibility of antibiotics.

Neurologic complications and sequellae of infectious diseases in Uganda and Kenya: Analysis of 288 cases from two rural hospitals.

OBJECTIVE: Many infections occurring in area of Sub-Saharan Africa are associated with more or less serious neurologic symptoms or complications. The aim of this study was to assess the incidence of selected infectious diseases in the equatorial part of Uganda and Kenya and to monitor potential neurological complications of these infections. METHODS: The study was performed for May - August 2008. Patients suffering from cerebral malaria, AIDS, meningitis, typhoid, tuberculosis (TB), syphilis, leprosy, and trypanosomiasis patients were enrolled. Besides of standard examination, lumbar puncture (LP) and cerebrospinal fluid (CSF) examination was performed, and the occurrence of neurological disorders and sequellae was recorded and assessed. RESULTS: Altogether 288 patients with neurological manifestation were enrolled. Malaria was the most prevalent disease in this study (102 cases, 35.42%), followed by typhoid (47 cases, 16.2%) and meningitis (38 cases, 13.2%). Leprosy and trypanosomiasis were only rarely detected (2.3% and 1.4%, respectively). CONCLUSION: In malaria and HIV hyper-endemic area of rural Uganda, cerebral malaria is the leading tropical neuroinfection. Also, meningitis is still frequent probably due to insufficient access to vaccination.

Numbers of non-severe as well as cerebral malaria cases are decreasing with higher altitude in Burundi Highlands, but still remains high.

OBJECTIVE: We aimed to assess the occurrence of malaria-positive cases in 4 rural Burundian hospitals during December 2011 placed at different altitudes above sea level. METHODS: Diagnosis of malaria was made upon considering clinical symptoms, microscopic evaluation and rapid diagnostic test results. We performed analysis of patient's clinical data collected in 4 hospitals in Burundi to compare the occurrence of malaria at different altitudes. RESULTS: The lowest incidence of malaria during December 2011 was detected at Murago Hospital (606 cases per month, 47.6%), which is located in the highest altitude, and the highest occurrence was in Gasura (1,559 cases, 91.3%), then in Rutovu (732 cases, 81.2%) and Buraniro (4,436 cases, 78.6%). Compared with other types of consultations (gynecological, HIV/AIDS, other tropical diseases), malaria was the most frequent reason for medical consultation. CONCLUSION: We have observed the lowest occurrence of malaria at hospitals located in the highest altitude. Despite the lower number of malaria cases in higher altitudes, its impact on public health should not be underestimated.

Susceptibility to caspofungin and fluconazole and Als1/Als3 gene expression in biofilm and dispersal cells of Candida albicans.

The biofilm of Candida albicans has been implicated as a source of bloodstream infections. Dispersal cells, as the final biofilm stage, are responsible for its spread. The aim of this study was to compare the susceptibility of biofilm and dispersal cells vs. planktonic cells (overnight liquid culture) of C. albicans to caspofungin (CAS) and fluconazole (FLU) when the drugs were added: i) at the beginning of the experiment; ii) after 1.5 h (adherence stage); iii) after 24 h (early mature biofilm). The findings were evaluated after 48 h (mature biofilm) using the XTT reduction assay. Later administration of the drug increased biofilm sessile minimal inhibitory concentration (SMIC(80)) of both FLU and CAS from 1 µg mL(-1) to over 64 µg mL(-1) and from 0.125 µg mL(-1) to over 16 µg mL(-1), respectively. Susceptibility of dispersal cells also decreased with time of administration. We also determined the expression of the Als1 and Als3 genes in 48-h sessile biofilm and dispersal cells of C. albicans SC5314 and compared it to planktonic cells. The expression was normalised to the standard Act1 gene in every condition tested. Quantitative real-time PCR revealed a strong up-regulation of the Als1 gene in the dispersal cells but not in biofilm and high expression of the Als3 gene in both biofilm and dispersal cells. High expression of both Als1 and Als3 genes supports the hypothesis that dispersal cells pose a high-risk of infection.

Cdr2p contributes to fluconazole resistance in Candida dubliniensis clinical isolates.

The development of resistance to azole antifungals used in the treatment of fungal infections can be a serious medical problem. Here, we investigate the molecular mechanisms associated with reduced susceptibility to fluconazole in clinical isolates of Candida dubliniensis , showing evidence of the trailing growth phenomenon. The changes in membrane sterol composition were studied in the presence of subinhibitory fluconazole concentrations. Despite lanosterol and eburicol accumulating as the most prevalent sterols after fluconazole treatment, these ergosterol precursors still support growth of Candida isolates. The overexpression of ABC transporters was demonstrated by immunoblotting employing specific antibodies against Cdr1p and Cdr2p. The presence of a full-length 170 kDa protein Cdr1p was detected in two isolates, while a truncated form of Cdr1p with the molecular mass of 85 kDa was observed in isolate 966/3(2). Notably, Cdr2p was detected in this isolate, and the expression of this transporter was modulated by subinhibitory concentrations of fluconazole. These results suggest that C. dubliniensis can display the trailing growth phenomenon, and such isolates express similar molecular mechanisms like that of fluconazole-resistant isolates and can therefore be associated with recurrent infections.