- Volume 64, Issue 5, 2015

Metals such as mercury, arsenic, copper and silver have been used in various forms as antimicrobials for thousands of years with until recently, little understanding of their mode of action. The discovery of antibiotics and new organic antimicrobial compounds during the twentieth century saw a general decline in the clinical use of antimicrobial metal compounds, with the exception of the rediscovery of the use of silver for burns treatments and niche uses for other metal compounds. Antibiotics and new antimicrobials were regarded as being safer for the patient and more effective than the metal-based compounds they supplanted. Bacterial metal ion resistances were first discovered in the second half of the twentieth century. The detailed mechanisms of resistance have now been characterized in a wide range of bacteria. As the use of antimicrobial metals is limited, it is legitimate to ask: are antimicrobial metal resistances in pathogenic and commensal bacteria important now? This review details the new, rediscovered and ‘never went away’ uses of antimicrobial metals; examines the prevalence and linkage of antimicrobial metal resistance genes to other antimicrobial resistance genes; and examines the evidence for horizontal transfer of these genes between bacteria. Finally, we discuss the possible implications of the widespread dissemination of these resistances on re-emergent uses of antimicrobial metals and how this could impact upon the antibiotic resistance problem.

Helicobacter pylori (H. pylori) may enter into host cells, maybe as a facultative intracellular pathogen. This study aims to reveal the roles of internalized H. pylori in the bacterial pathopoiesis. Transmission electron microscopy was used to observe the invasion of H. pylori. Invasion rates of H. pylori (two standard strains and 43 clinical strains) were examined by gentamicin invasion assay. The cagA, cagE and vacA genes of H. pylori were detected by PCR. The cagA 3′region (cagA-EPIYA) of each strain was sequenced. The secretion of IL-8 from AGS cells and activity of NF-κB induced by intracellular H. pylori were tested by ELISA and the dual-luciferase reporter assay system, respectively. It was found that H. pylori could adhere to and invade AGS cells, then continue to survive and multiply in the cytoplasm. The average invasion rate of H. pylori gastric cancer plants and that of ulcer plants were both higher than that of gastritis plants (P≈0.0001). In the clinical strains, cagA, vacA and cagE were all positive; cagA-EPIYA genotypes included ABD 90.7 % (39/43) and ABBD 9.3 % (4/43), all without comparability. Notably, the average invasion rate of H. pylori vacA s1c-i1-m1b plants was higher than that of vacA s1c-i1-m2 plants (P = 0.0445). In addition, the intracellular H. pylori all could induce IL-8 secretion, which was decreased after cells were pretreated with anti-β1-integrin antibody or SN-50 (an NF-κB inhibitor). The intracellular H. pylori all activated NF-κB, which would be inhibited after cells were pretreated with anti-β1-integrin antibody. These results demonstrate that H. pylori invasive ability and disease severity have a positive correlation, and this intension of invasive ability is associated with the vacA mid-region, not with cagA, cagA-EPIYA or cagE. It is possible that cagA and cagE are essential for the bacterial invasion. Internalized H. pylori can activate NF-κB signal pathway and induce IL-8 secretion, which suggests that H. pylori invasion may be an important strategy to play a role in the development of H. pylori associated diseases.

Considering the prevalence of Leptospira santarosai infections in the Americas and the scarce information about the species, we aimed to apply a multilocus variable number tandem repeat (VNTR) analysis (MLVA) for the molecular typing of L. santarosai isolates from various sources. Amplification of three VNTR loci selected from L. santarosai genome sequences resulted in a wide range of sizes for the amplified products amongst the 21 L. santarosai strains analysed. This suggested a variation in tandem repeat copy numbers in the VNTR loci. secY sequencing also showed a high nucleotide diversity, confirming the MLVA data. In conclusion, this novel MLVA provided a high level of discrimination between L. santarosai isolates, and this new typing tool could be used to investigate leptospirosis in regions where L. santarosai predominates.

Minocycline (MIN) and tigecycline (TIG) are antibiotics currently used for treatment of multidrug-resistant nosocomial pathogens. In this work, we show that blue light, as well as white light, modulates susceptibility to these antibiotics in a temperature-dependent manner. The modulation of susceptibility by light depends on the content of iron; an increase in iron results in a reduction in antibiotic susceptibility both under light and in the dark, though the effect is more pronounced in the latter condition. We further provide insights into the mechanism by showing that reduction in susceptibility to MIN and TIG induced by light is likely triggered by the generation of 1O2, which, by a yet unknown mechanism, would ultimately lead to the activation of resistance genes such as those coding for the efflux pump AdeABC. The clinical relevance of these results may lie in surface-exposed wound infections, given the exposure to light in addition to the relatively low temperatures recorded in this type of lesion. We further show that the modulation of antibiotic susceptibility occurs not only in Acinetobacter baumannii but also in other micro-organisms of clinical relevance such as Escherichia coli and Staphylococcus aureus. Overall, our findings allow us to suggest that MIN and TIG antibiotic treatments may be improved by the inclusion of an iron chelator, in addition to keeping the wounds in the dark, a condition that would increase the effectiveness in the control of infections involving these micro-organisms.

Clinical isolates of Klebsiella pneumoniae and Klebsiella oxytoca collected from 20 Japanese medical facilities between 2000 and 2010 were analysed to evaluate the mechanisms of resistance and antibacterial susceptibilities to 14 antimicrobials. Overall, eight of 484 (1.6 %) K. pneumoniae and 19 of 359 (5.3 %) K. oxytoca were determined to be extended-spectrum β-lactamase (ESBL) phenotype isolates, and the identified ESBLs amongst the K. pneumoniae isolates were CTX-M-2, -3, -14 and -15, and SHV-12. In contrast, overproduction of chromosomal β-lactamase OXY-2, which was due to a distinct mutation at the − 10 promoter region of this gene, conferred the ESBL phenotype to all the K. oxytoca isolates except one. Based on the Clinical and Laboratory Standards Institute breakpoints, all the ESBL phenotype K. pneumoniae were susceptible to doripenem, flomoxef, moxalactam (latamoxef), cefmetazole and tazobactam/piperacillin, whereas the ESBL phenotype K. oxytoca were susceptible to ceftazidime and ceftibuten in addition to the above, with the exception of tazobactam/piperacillin. Amongst the oral antimicrobials, ceftibuten was relatively effective against both ESBL phenotype Klebsiella species compared with levofloxacin and amoxicillin/clavulanic acid.

Norovirus is the leading cause of viral gastroenteritis globally. Norovirus genotype GII.4 is responsible for the majority of outbreaks, but new variants are continuously emerging. The objective of the study was to delineate the clinical manifestations and complications associated with these new norovirus GII.4 variants in children. We investigated norovirus infections from the community outbreak in October 2011–September 2012 and an earlier outbreak in 2006–2007, in northern Taiwan. Norovirus genotypes and their variants were validated using molecular methods. A norovirus outbreak started in mid-2011 and continued through 2012 in northern Taiwan. Hospitalized children infected by norovirus in 2012 showed a significantly higher incidence of intestinal haemorrhage, as indicated by grossly bloody faeces (P = 0.012) and occult blood in faeces (P <0.001), and also presented with more high fever >39 °C (P <0.001), fever >38.5 °C (P <0.001) and fever of any temperature >38 °C (P <0.001), compared with children hospitalized in 2006–2007. Analysis of 20 near-full-length genome sequences indicated an emergence of GII.4 2012 variants in 2011–2012. Circulating noroviruses can be divided into two clusters: GII.4 2012a, which is identical to the newly reported strain GII.4 Sydney 2012, and GII.4 2012b, which is close to GII.4 2006b, the earlier predominant strain. The emerging new variants of norovirus GII.4 caused a distinct clinical syndrome of acute gastroenteritis with severe fever and a high rate of intestinal haemorrhage in children. The genetic diversity associated with changing clinical manifestations poses major obstacles to norovirus control.

Chlamydia psittaci is prevalent in broiler chicken production. However, the role of C. psittaci in the respiratory disease complex needs to be clarified. Our aim was to identify the time point when a C. psittaci infection appeared on a broiler farm and to examine the presence of other respiratory pathogens at that time. We focused on the ‘major’ respiratory pathogens occurring in Belgian broilers, namely infectious bronchitis virus (IBV), avian metapneumovirus (aMPV), Ornithobacterium rhinotracheale, Mycoplasma gallisepticum and Mycoplasma synoviae, and examined their co-occurrence with C. psittaci on three commercial broiler farms. For all farms, 1-day-old broilers showed high maternal antibody titres against C. psittaci in the presence of viable C. psittaci. Maternal antibodies seemed to protect against respiratory signs. Maternal antibodies declined and clinical outbreaks could be identified serologically even before maternal antibodies completely disappeared. Mixed infections with genotypes B/C and B/C/D were observed. Broilers with C. psittaci antibody increases showed conjunctivitis, signs of upper respiratory disease and dyspnoea. C. psittaci always preceded an O. rhinotracheale infection. Infections with aMPV, IBV or Mycoplasma spp. were not observed. Evidence was provided that C. psittaci could occur at an early age in broilers without a predisposing respiratory infection. Both C. psittaci and O. rhinotracheale should be considered when developing prevention strategies for respiratory disease in broilers.