Estimating Bordetella pertussis seroprevalence in adolescents and young adults in Mexico using the 2012 National Health and Nutrition Survey (ENSANUT).

BACKGROUND: Low vaccination rates and under-detection of pertussis infections in adolescents and young adults have an impact on the transmission of pertussis to infants. In this study, the proportion of adolescents and young adults with IgG antibodies against B. pertussis antigens, representing recent infection or vaccination, was estimated in a population-based probabilistic survey in Mexico. METHODS: Sera and data from 1,581 subjects, including 1,102 adolescents and 479 young adults (10-19 and 20-25 years old, respectively) randomly selected from Mexico's 2012 National Health and Nutrition Survey, were analyzed. IgG antibodies against pertussis toxin (PT) were measured with the CDC/FDA ELISA. A subset of 234 samples was additionally tested with Bp-IgG PT ELISA kit (EUROIMMUN AG, Lubeck, Germany). Threshold values from corresponding test kits were used to identify recent infection or vaccination. RESULTS: Overall anti-PT IgG seroprevalence was 3.9% (95% CI: 2.3-6.3); 3.1% (95% CI: 1.9-5.0) in adolescents, and 4.9% (95% CI: 2.2-11) in young adults. Seroprevalence did not significantly vary by sex, socioeconomic status, region or rural/urban location. Compared to the CDC/FDA ELISA, the EUROIMMUN test showed a 76% sensitivity and 88% specificity. The weighted estimates represent a considerable burden of recent infection in adolescents and young adults; however, most adolescents and adults were seronegative and, therefore, susceptible to pertussis infection. CONCLUSION: Since booster vaccination to B. pertussis after toddlerhood is not recommended in the Mexican national policy, anti-PT IgG seropositivity may be reasonably attributed to recent infection. Assessing pertussis seroprevalence requires careful consideration of the diagnostic test threshold interpretation and epidemiological model used.

Acinetobacter baumannii infections in a tertiary care hospital in Mexico over the past 13 years.

BACKGROUND: Acinetobacter baumannii has evolved from an opportunistic pathogen into a common and persistent nosocomial bacterium capable of causing severe infections during endemic and epidemic periods. METHODS: The study period extended from January 1999 to December 2011 and involved patients hospitalized at the Hospital Civil de Guadalajara, Fray Antonio Alcalde, Jalisco, Mexico. From each patient, a single isolate was obtained, and a total of 3,680 unique isolates were collected. Susceptibility tests were performed according to the guidelines of the Clinical and Laboratory Standards Institute. RESULTS: A. baumannii has disseminated throughout the Hospital Civil de Guadalajara, Fray Antonio Alcalde, since 1999. A. baumannii isolates obtained from patients treated in the adult intensive care unit represent the majority of the isolates that have been collected. In addition, A. baumannii was isolated from the adult neurosurgical ward and the adult internal medicine ward, and these isolates were frequently obtained from secretions. A persistent decrease in the susceptibility of A. baumannii isolates to meropenem (92% in 1999 to 12% in 2011), imipenem and amikacin has been observed. CONCLUSIONS: A. baumannii became an endemic nosocomial pathogen during the study period at the Hospital Civil de Guadalajara, Fray Antonio Alcalde, and has exhibited a persistent decrease in susceptibility to all categories of antimicrobial agents over the past 13 years.

Non-typhi Salmonella serovars found in Mexican zoo animals.

The aim of the present study was to determine the bacteriological prevalence of subclinical non-typhi Salmonella infections in zoo animals and to determine the most frequently isolated serovars of the bacteria. A total of 267 samples were analyzed, including fecal samples from zoo animals and rodents, insects (Musca domestica and Periplaneta americana) and samples of the zoo animal's food. Salmonella was detected in 11.6% of the samples analyzed. Characterization of the isolates was performed with serotyping and pulsed-field gel electrophoresis. The following serovars were isolated: S. San Diego, S. Oranienburg, S. Weltevreden, S. Braenderup, S. Derby, S. 6,7, H:en x:- and S. 3,10, H:r:-. The isolates showed seven pulsed-field gel electrophoresis patterns with a Jaccard coefficient≥0.75 indicating a possible common origin. The prevalence of asymptomatic infections caused by Salmonella spp. in zoo animals was high. These findings demonstrate the diversity of Salmonella serovars in several captive wild animal species.

The blaSHV-5 gene is encoded in a compound transposon duplicated in tandem in Enterobacter cloacae.

The presence of bla(SHV-5) is described in a compound transposon, duplicated in tandem and flanked by IS26 copies on a 70-kb conjugative plasmid (pHNM1), in an Enterobacter cloacae strain associated with a nosocomial outbreak that occurred in Mexico.

Lipopopeptidephosphoglycan from Entamoeba histolytica activates human macrophages and dendritic cells and reaches their late endosomes.

Lipopopeptidephosphoglycan (LPPG) is a complex macromolecule from the surface of Entamoeba histolytica trophozoites. We analysed the interaction between LPPG and human macrophages and dendritic cells (DCs) and found that LPPG is internalized by these cells and activates them. The internalization process involves intracellular traffic from the cell membrane to late endosomes, as shown by co-localization of LPPG with late endosomes marked with FITC-dextran and LAMP-1. LPPG-activated DCs have increased expression of co-stimulatory molecules CD80, CD86 and CD40 and produce pro-inflammatory cytokines TNF-alpha, IL-8 and IL-12. Taken together, these results show that LPPG activates antigen-presenting cells and reaches intracellular compartments that are involved in antigen presentation.

Multiresistant extended-spectrum beta-lactamase-producing Klebsiella pneumoniae causing an outbreak of nosocomial bloodstream infection.

This article describes an outbreak of bloodstream infection due to clonal dissemination of multiresistant Klebsiella pneumoniae in a neonatal area, during August 1999, in Mexico City General Hospital. The intestinal tract was the likely reservoir, and intensification of Contact Precaution measures contained the outbreak.

Outbreak of nosocomial sepsis and pneumonia in a newborn intensive care unit by multiresistant extended-spectrum beta-lactamase-producing Klebsiella pneumoniae: high impact on mortality.

We describe a case-control study of a small outbreak of nosocomial sepsis and pneumonia with high mortality due to clonal dissemination of a multiresistant Klebsiella pneumoniae in the neonatal intensive care unit of a Mexican institution. Our study helped to change nosocomial infection control policy in this hospital.

Rapid and complete fusion of macrophage lysosomes with phagosomes containing Salmonella typhimurium.

The virulence of Salmonella typhimurium for mice results, in part, from its ability to survive after phagocytosis by macrophages. Although it is generally agreed that intracellular bacteria persist in membrane-bound phagosomes, there remains some question as to whether these phagosomes fuse with macrophage lysosomes. This report describes the maturation of phagosomes containing S. typhimurium inside mouse bone marrow-derived macrophages. Macrophages were infected briefly and incubated for various intervals; then they were examined by fluorescence microscopy for colocalization of bacteria with lysosomal markers. These markers included LAMP-1, cathepsin L, and fluorescent proteins or dextrans preloaded into lysosomes by endocytosis. By all measures, phagosomes containing S. typhimurium merged completely with the lysosomal compartment within 20 min of phagocytosis. The rate of phagosome-lysosome fusion was similar to the rate for phagocytosed latex beads. Phagolysosomes remained accessible to fluid-phase probes and contained lysosomal markers for many hours. Moreover, a large percentage of the wild-type bacteria that were viable 20 min after infection survived longer incubations inside macrophages, indicating that the survivors were not a minor subpopulation that avoided phagosome-lysosome fusion. Therefore, we conclude that S. typhimurium survives within the lysosomal compartments of macrophages.

Spacious phagosome formation within mouse macrophages correlates with Salmonella serotype pathogenicity and host susceptibility.

Light microscopic studies indicated a correlation between the virulence for mice of different Salmonella serotypes and the ability to form or maintain spacious phagosomes (SP) within mouse macrophages. Although Salmonella typhimurium induced membrane ruffling, macropinocytosis, and SP formation in macrophages from BALB/c mice, serotypes which are nonpathogenic for mice produced markedly fewer SP. SP formation correlated with both serotype survival within mouse macrophages and reported lethality for mice. Time-lapse video microscopy demonstrated that the human pathogen S. typhi induced generalized macropinocytosis and SP formation in human monocyte-derived macrophages, indicating a similar morphology for the initial phases of this host-pathogen interaction. In contrast to bone marrow-derived macrophages from BALB/c mice, macrophages from S. typhimurium-resistant outbred (CD-1) and inbred (CBA/HN) mice did not initiate generalized macropinocytosis after bacterial infection and formed markedly fewer SP. These deficiencies were not due to the Ihy resistance genotype of these mice, as macrophages from mice that were congenic except for the Ihy locus demonstrated equal SP formation in response to S. typhimurium. The observation that S. typhimurium-resistant CD-1 and CBA/HN mice are deficient in the ability to form and/or maintain SP indicates that a variable host component is important for SP formation and suggests that the ability to induce or form SP affects susceptibility to S. typhimurium. When serotypes nonpathogenic for mice were used to infect BALB/c macrophages, or when CD-1 or CBA/HN mouse macrophages were infected by S. typhimurium, some of the SP that formed shrank within seconds. This rapid shrinkage suggests that SP maintenance is also important for S. typhimurium survival within macrophages. These studies indicate that both host and bacterial factors contribute to SP formation and maintenance, which correlate with Salmonella intracellular survival and the ability to cause lethal enteric (typhoid) fever.

Characterization of the Salmonella typhimurium pagC/pagD chromosomal region.

The PhoP/PhoQ two-component system regulates Salmonella typhimurium genes that are essential to bacterial virulence and survival within macrophages. The best characterized of these PhoP-activated genes (pag) is pagC, which encodes a 188-amino-acid envelope protein (W. S. Pulkkinen and S. I. Miller, J. Bacteriol. 173:86-93, 1991). We here report the identification of four genes (pagD, envE, msgA, and envF) located 5' to pagC. Each gene is transcribed from its own promoter, two of which (msgA and pagD) were defined by primer extension analysis. Three of these genes (pagD, envE, and envF) are predicted to encode envelope proteins. The pagD gene is transcribed in a direction opposite from that of and adjacent to pagC and is positively regulated by PhoP/PhoQ. Transposon insertions within pagD and msgA attenuate bacterial virulence and survival within macrophages; however, deletion of pagD has no effect on virulence. The product of the envF gene is predicted to be a lipoprotein on the basis of the presence of a consensus lipid attachment site. The low G + C content of these genes and the homology of msgA to Shigella plasmid DNA suggest that this region may have been acquired by horizontal transmission.

Salmonella stimulate macrophage macropinocytosis and persist within spacious phagosomes.

Light microscopic studies of phagocytosis showed that Salmonella typhimurium entered mouse macrophages enclosed in spacious phagosomes (SP). Viewed by time-lapse video microscopy, bone marrow-derived macrophages exposed to S. typhimurium displayed generalized plasma membrane ruffling and macropinocytosis. Phagosomes containing Salmonella were morphologically indistinguishable from macropinosomes. SP formation was observed after several methods of bacterial opsonization, although bacteria opsonized with specific IgG appeared initially in small phagosomes that later enlarged. In contrast to macropinosomes induced by growth factors, which shrink completely within 15 min, SP persisted in the cytoplasm, enlarging often by fusion with macropinosomes or other SP. A Salmonella strain containing a constitutive mutation in the phoP virulence regulatory locus (PhoPc) induced significantly fewer SP. Similar to Yersinia enterocolitica, PhoPc bacteria entered macrophages in close-fitting phagosomes, consistent with that expected for conventional receptor-mediated phagocytosis. These results suggest that formation of SP contributes to Salmonella survival and virulence.

The PhoP virulence regulon and live oral Salmonella vaccines.

The PhoP virulence regulon is essential to Salmonella typhimurium mouse typhoid fever pathogenesis and survival within macrophages. This virulence regulon is composed of the PhoP (transcriptional regulator) and PhoQ (environmental sensor) proteins and the genetic loci they positively (pags for PhoP activated genes) and negatively (prgs for PhoP repressed genes) regulate. Three regulated loci pagC, pagD, and prgH, when singly mutated, affect the virulence of S. typhimurium for mice. Strains with phoP locus mutations are effective as live vaccines in mice, and strains with a constitutive regulatory mutation, a point mutation in PhoQ, can protect mice against typhoid fever when only very few organisms are administered. The addition of various PhoP regulon mutations further attenuates aroA mutants of S. typhimurium, suggesting that these mutations would be useful in further attenuating vaccine strains with metabolic pathway mutations. The phoP, phoQ, pagC, and pagD genes are highly conserved between S. typhimurium and S. typhi and may be valuable as mutations in live vaccines for human typhoid fever. A plasmid suicide vector that allows deletion of the pagC gene and stable insertion of heterologous antigen genes within the deleted pagC locus has been constructed and used successfully in S. typhimurium and S. typhi.

Salmonella typhimurium activates virulence gene transcription within acidified macrophage phagosomes.

Survival of Salmonella typhimurium within macrophage phagosomes requires the coordinate expression of bacterial gene products. This report examines the contribution of phagosomal pH as a signal for expression of genes positively regulated by the S. typhimurium virulence regulators PhoP and PhoQ. Several hours after bacterial phagocytosis by murine bone marrow-derived macrophages, PhoP-activated gene transcription increased 50- to 77-fold. In contrast, no difference in PhoP-activated gene expression was observed after infection of cultured epithelial cells, suggesting that the membrane sensor PhoQ recognized signals unique to macrophage phagosomes. The increase in PhoP-regulated gene expression was abolished when macrophage culture medium contained NH4Cl or chloroquine, weak bases that raise the pH of acidic compartments. Measurements of pH documented that S. typhimurium delayed and attenuated acidification of its intracellular compartment. Phagosomes containing S. typhimurium required 4-5 hr to reach pH < 5.0. In contrast, within 1 hr vacuoles containing heat-killed bacteria were measured at pH < 4.5. The eventual acidification of phagosomes to pH < 5.0 correlated with the period of maximal PhoP-dependent gene expression. These observations implicate phagosome acidification as an intracellular inducer of PhoP-regulated gene expression and suggest that Salmonella survival is dependent on its ability to attenuate phagosome acidification.

[Antimicrobial sensitivity profile of Staphylococcus aureus at a pediatric hospital: prevalence of resistance to methicillin]. / Patrón de sensibilidad antimicrobiana de Staphylococcus aureus en un hospital pediátrico: prevalencia de resistencia a meticilina.

Methicillin-resistant Staphylococcus aureus (MRSA) has become a significant cause of nosocomial infections. In efforts to delineate the magnitude of this problem, we determined the prevalence of MRSA in community acquired (n = 382) and nosocomial strains (n = 207) of S. aureus isolated between Jan 1986 and March 1989. Antimicrobial susceptibility was evaluated using an agar dilution method (Muller-Hinton agar supplemented with 4% NaCl incubated to 35 degrees C for 24 h) and MIC breakpoints were determined according to NCCLS standards. We detected (24.2%) MRSA in nosocomial strains and (5%) MRSA in community acquired strains (P less than 0.05), with a global prevalence of 11.7%. The susceptibility of community acquired S. aureus was 90% or higher for dicloxacillin, cephalothin, sulbactam/ampicillin (S/A), clindamycin, rifampicin and amikacin; 85% for cefotaxime and SMX/TMP and only 75% for erythromycin. The susceptibility pattern of the nosocomial strains was consistent with the prevalence of MRSA but the susceptibility for cephalothin, amikacin and sulbactam/ampicillin was 84.4%, 89.4% and 86.5% respectively, significantly higher than for methicillin (P less than 0.05). Although the increased susceptibility for cephalothin and amikacin has been reported for MRSA before, the published reports using these antibiotics in the treatment of MRSA infections are controversial. The increased susceptibility of MRSA to S/A could be explained in part if the MR was mediated by "acquired MR" attributable to B-lactamase production. Our data provide a perspective on the magnitude of MRSA as a problem in a pediatric teaching hospital in Mexico. Moreover, if taken at face value, the in vitro susceptibility data point to various potential treatment options which warrant clinical evaluation.