Bordetella bronchiseptica Diguanylate Cyclase BdcA Regulates Motility and Is Important for the Establishment of Respiratory Infection in Mice.

Bacteria can be motile and planktonic or, alternatively, sessile and participating in the biofilm mode of growth. The transition between these lifestyles can be regulated by a second messenger, cyclic dimeric GMP (c-di-GMP). High intracellular c-di-GMP concentration correlates with biofilm formation and motility inhibition in most bacteria, including Bordetella bronchiseptica, which causes respiratory tract infections in mammals and forms biofilms in infected mice. We previously described the diguanylate cyclase BdcA as involved in c-di-GMP synthesis and motility regulation in B. bronchiseptica; here, we further describe the mechanism whereby BdcA is able to regulate motility and biofilm formation. Amino acid replacement of GGDEF with GGAAF in BdcA is consistent with the conclusion that diguanylate cyclase activity is necessary for biofilm formation and motility regulation, although we were unable to confirm the stability of the mutant protein. In the absence of the bdcA gene, B. bronchiseptica showed enhanced motility, strengthening the hypothesis that BdcA regulates motility in B. bronchiseptica We showed that c-di-GMP-mediated motility inhibition involved regulation of flagellin expression, as high c-di-GMP levels achieved by expressing BdcA significantly reduced the level of flagellin protein. We also demonstrated that protein BB2109 is necessary for BdcA activity, motility inhibition, and biofilm formation. Finally, absence of the bdcA gene affected bacterial infection, implicating BdcA-regulated functions as important for bacterium-host interactions. This work supports the role of c-di-GMP in biofilm formation and motility regulation in B. bronchiseptica, as well as its impact on pathogenesis.IMPORTANCE Pathogenesis of Bordetella spp., like that of a number of other pathogens, involves biofilm formation. Biofilms increase tolerance to biotic and abiotic factors and are proposed as reservoirs of microbes for transmission to other organs (trachea, lungs) or other hosts. Bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) is a second messenger that regulates transition between biofilm and planktonic lifestyles. In Bordetella bronchiseptica, high c-di-GMP levels inhibit motility and favor biofilm formation. In the present work, we characterized a B. bronchiseptica diguanylate cyclase, BdcA, which regulates motility and biofilm formation and affects the ability of B. bronchiseptica to colonize the murine respiratory tract. These results provide us with a better understanding of how B. bronchiseptica can infect a host.

CCL28 modulates neutrophil responses during infection with mucosal pathogens.

The chemokine CCL28 is highly expressed in mucosal tissues, but its role during infection is not well understood. Here, we show that CCL28 promotes neutrophil accumulation in the gut of mice infected with Salmonella and in the lung of mice infected with Acinetobacter. Neutrophils isolated from the infected mucosa expressed the CCL28 receptors CCR3 and, to a lesser extent, CCR10, on their surface. The functional consequences of CCL28 deficiency varied between the two infections: Ccl28-/- mice were highly susceptible to Salmonella gut infection but highly resistant to otherwise lethal Acinetobacter lung infection. In vitro, unstimulated neutrophils harbored pre-formed intracellular CCR3 that was rapidly mobilized to the cell surface following phagocytosis or inflammatory stimuli. Moreover, CCL28 stimulation enhanced neutrophil antimicrobial activity, production of reactive oxygen species, and formation of extracellular traps, all processes largely dependent on CCR3. Consistent with the different outcomes in the two infection models, neutrophil stimulation with CCL28 boosted the killing of Salmonella but not Acinetobacter. CCL28 thus plays a critical role in the immune response to mucosal pathogens by increasing neutrophil accumulation and activation, which can enhance pathogen clearance but also exacerbate disease depending on the mucosal site and the infectious agent.

High Prevalence of CTX-M-1-Like Enzymes in Urinary Isolates of Escherichia coli in Guayaquil, Ecuador.

Escherichia coli is one of the major causes of urinary tract infections in primary healthcare, and treatment is more complicated due to the increase in antibiotic resistance. Extended-spectrum ß-lactamases are the most common mechanism of resistance against third-generation cephalosporin, and CTX-M-like are among the most prevalent. The aim of our work is to investigate the prevalence of blaCTX-M in isolates of E. coli obtained from samples of patients without previous known contact with the hospital. Ninety-four E. coli isolates with resistance to third-generation cephalosporin were collected between 2008 and 2013 in Guayaquil, Ecuador. Polymerase chain reaction, followed by sequencing, was performed to identify the type of blaCTX-M-Like. Enterobacterial repetitive intergenic consensus (ERIC)-PCR was carried out to determine the clonal relationship between isolates. These results show an increase in resistance to third-generation cephalosporin from 10.58% to 23.96%. CTX-M-15 was the most prevalent mechanism of resistance being that the isolates were not clonal. Overall, these results show an increase in antibiotic resistance in the community over time, suggesting that more precise antibiotic stewardship needs to be implemented to control the dissemination of antibiotic-resistant bacteria in this region.

Early detection and control of an Acinetobacter baumannii multi-resistant outbreak in a hospital in Quito, Ecuador.

INTRODUCTION: Acinetobacter baumannii (ABA) is an important opportunistic pathogen associated with high mortality rates in intensive care units (ICUs). An outbreak in the ICU of a secondary-level hospital in Quito, Ecuador, occurred during April and May 2015 and was successfully controlled. METHODOLOGY: Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) and repetitive element palindromic (REP)-PCR was conducted on all isolates recovered from patients, as well as environmental samples, to confirm the presence of an outbreak. A case-control study was conducted by comparing the clinical histories of the affected patients and of control patients present in the ICU during the outbreak period who did not present a positive culture for ABA. RESULTS: Five patients were infected and two were colonized with the same clonal strain of ABA, which was also identified on the stethoscope and a monitor associated with an isolation room. Statistical analysis of case histories did not identify any additional risk factors, but the outbreak was initiated by one patient in the isolation room of the ICU who was infected with the outbreak strain. All patients who ocupied that room after the index case tested positive for at least one culture of ABA. The outbreak strain was found on the stethoscope, and a subclone was found on the monitor of that room. CONCLUSION: Having access to basic equipment will enable well-trained professionals to rapidly detect and initiate the control process of an outbreak, saving lives and money spent on nosocomial infection treatments.

Epidemiology of Tropical Neglected Diseases in Ecuador in the Last 20 Years.

BACKGROUND: Tropical and zoonotic diseases are major problems in developing countries like Ecuador. Poorly designed houses, the high proportion of isolated indigenous population and under developed infrastructure represent a fertile environment for vectors to proliferate. Control campaigns in Ecuador over the years have had varying success, depending on the disease and vectors targeted. AIMS: In our study we analyse the current situation of some neglected diseases in Ecuador and the efficiency of the control campaigns (by measuring changes in numbers of cases reported) that the Ecuadorian government has been running to limit the spread of these infectious and parasitic diseases. RESULTS: Our study reveals that Brucellosis, Chagas Disease, Rabies and Onchocerciasis have been controlled, but small outbreaks are still detected in endemic areas. Leptospirosis and Echinococcosis have been increasing steadily in recent years in Ecuador since the first records. The same increase has been reported world-wide also. Better diagnosis has resulted in a higher number of cases being identified, particularly with regard to the linking of outdoor activities and contact with farm animals as contributing vectors. Improvements in diagnosis are due to regular professional training, implementation of automatized systems, establishing diagnosis protocols and the creation of an epidemiological vigilance network that acts as soon as a case is reported. CONCLUSION: Control campaigns performed in Ecuador have been successful in recent years, although natural phenomena limit their efficiency. Leptospirosis and Echinococcosis infections remain a growing problem in Ecuador as it is worldwide.