Search

Bartonella species are fastidious Gram negative vector-borne bacteria with a wide range of mammalian reservoirs. While it is understood that some species Bartonella are human pathogens, the extent of human exposure to Bartonella species (both pathogenic and non-pathogenic) has yet to be fully understood. To this end, residual sera from participants enrolled in undifferentiated fever studies in Cambodia, Ghana, Laos, and Peru were screened for the presence of IgG antibodies against B. quintana and B. henselae, using the FOCUS diagnostics Dual Spot- Bartonella IgG Immunofluorescence assay. Forty-eight patients with suspected or confirmed B. bacilliformis exposure or infection in Peru, were screened to assess cross-reactivity of the FOCUS assay for IgG against other Bartonella species. Ten of 13 patients with confirmed B. bacilliformis infection were Bartonella-specific IgG positive and overall, 36/48 of the samples were positive. Additionally, 79/206, 44/200, 101/180, and 57/100 of samples from Peru, Laos, Cambodia, and Ghana, respectively were Bartonella-specific IgG positive. Further, ectoparasites pools from Cambodia, Laos, and Peru were tested using quantitative real-time PCR (qPCR) for the presence of Bartonella DNA. Of the sand-fly pools collected in Peru, 0/196 were qPCR positive; 15/140 flea pools collected in Cambodia were qPCR positive; while 0/105 ticks, 0/22 fleas, and 0/3 louse pools collected in Laos tested positive for Bartonella DNA. Evidence of Bartonella in fleas from Cambodia supports the possibility that humans are exposed to Bartonella through this traditional vector. However, Bartonella species were not found in fleas, ticks, or lice from Laos, or sandflies from Peru. This could account for the lower positive serology among the population in Laos and the strictly localized nature of B. bacillformis infections in Peru. Human exposure to Bartonella species and Bartonella as a human pathogen warrants further investigation.

Rickettsia and Leptospira spp. are under-recognized causes of acute febrile disease worldwide. Rickettsia species are often placed into the spotted fever group rickettsiae (SFGR) and typhus group rickettsiae (TGR). We explored the antibody prevalence among humans for these two groups of rickettsiae in four regions of Peru (Lima, Cusco, Puerto Maldonado, and Tumbes) and for Leptospira spp. in Puerto Maldonado and Tumbes. We also assessed risk factors for seropositivity and collected serum samples and ectoparasites from peri-domestic animals from households in sites with high human seroprevalence. In total, we tested 2,165 human sera for antibodies (IgG) against SFGR and TGR by ELISA and for antibodies against Leptospira by a microscopic agglutination test. Overall, human antibody prevalence across the four sites was 10.6% for SFGR (ranging from 6.2% to 14.0%, highest in Tumbes) and 3.3% for TGR (ranging from 2.6% to 6.4%, highest in Puerto Maldonado). Factors associated with seroreactivity against SFGR were male gender, older age, contact with backyard birds, and working in agriculture or with livestock. However, exposure to any kind of animal within the household decreased the odds ratio by half. Age was the only variable associated with higher TGR seroprevalence. The prevalence of Leptospira was 11.3% in Puerto Maldonado and 5.8% in Tumbes, with a borderline association with keeping animals in the household. We tested animal sera for Leptospira and conducted polymerase chain reaction (PCR) to detect Rickettsia species among ectoparasites collected from domestic animals in 63 households of seropositive participants and controls. We did not find any association between animal infection and human serostatus.

Drinking water contamination is a frequent problem in developing countries and could be associated with bacterial pathogen carriage in feces. We evaluated the association between the risk of drinking water and bacterial carrier status in children younger than 5 years in a cross-sectional study conducted in 199 households from three Peruvian rural communities. Fecal samples from children were screened for pathogenic Aeromonas, Campylobacter, and Vibrio species, as well as for Enterobacteriaceae, including pathogenic Escherichia coli. The drinking water risk was determined using E. coli as an indicator of contamination. Nineteen (9.5%) children were colonized with pathogens and classified as carriers, all without diarrhea symptoms. Of 199 drinking water samples, 38 (19.1%) were classified as very high risk because of high fecal contamination (> 100 E. coli/100 mL). Shared-use water sources, daily washing of containers, and washing using only water were associated with higher prevalence of bacterial carriage, whereas there was no association between households reporting boiling and chlorination of water and carrier status. The prevalence of carriage in children exposed to very high-risk water was 2.82 (95% CI: 1.21-6.59) times the prevalence of those who consumed less contaminated water, adjusted by the water source and daily washing. Our results suggest that household drinking water plays an important role in the generation of carriers with diarrheal pathogens. Our findings also highlight the importance of interventions to ensure the safety of drinking water. Further studies are needed to validate the observed association and determine its significance with respect to diarrhea in the community.

Leptospirosis is a neglected zoonotic disease with worldwide endemicity and continues to be a significant public health burden on resource-limited populations. Previously, we produced three highly purified recombinant antigens (rLipL32, rLipL41, and rLigA-Rep) and evaluated their performance of detecting Leptospira-specific antibodies in enzyme-linked immunosorbent assay (ELISA) as compared with the microscopic agglutination test (MAT). The overall sensitivity of this assay approached 90%. Recently, another recombinant antigen (rLigB-Rep) was prepared. We tested each individual antigen and a 1:1:1:1 mixture of these four antigens for the detection of Leptospira-specific antibodies in ELISA. The performance of these recombinant antigens was evaluated with a much larger febrile patient panel (337 MAT-confirmed positive sera and 92 MAT-negative sera from febrile patients). Combining the detection results of immunoglobulin M and immunoglobulin G from these four individual antigens, the overall sensitivity was close to 90% but the specificity was only 66%, based on the MAT reference method. The overall sensitivity and specificity of the four-antigen mixture were 82% and 86%, respectively. The mixture of four antigens also exhibited a broader reactivity with MAT-positive samples of 18 serovars from six major pathogenic Leptospira species. Given the limitations of MAT, the data were further analyzed by Bayesian latent class model, showing that ELISA using a 1:1:1:1 mixture still maintained high sensitivity (79%) and specificity (88%) as compared with the sensitivity (90%) and specificity (83%) of MAT. Therefore, ELISA using a mixture of these four antigens could be a very useful test for seroprevalence studies.

Health-care–associated infections (HAI) and antimicrobial resistance are of great public health concern worldwide.1 Bacterial pathogens causing HAI have become increasingly more resistant over the past 10–15 years as a result of various mechanisms, including gene-mediated enzymes such as class A, B, and D β-lactamases.2 Numerous factors, from the horizontal transfer of gene-encoded enzymes to global travel, have allowed resistance and resistant organisms to rapidly spread with great clinical and epidemiological impact.3,4 The recently described carbapenemase, New Delhi metallo-β-lactamase (NDM-1) class B, is of particular epidemiological and clinical concern. Since its discovery in 2008, NDM-1 has rapidly spread worldwide and confers resistance to almost all lactams, with the exception of aztreonam, leaving limited therapeutic options against pathogens harboring NDM-1, typically colistin, tigecycline, and fosfomycin.2,5,6 In Latin America, the blaNDM-1 gene was first reported in 2011 from Guatemala and Colombia, and later from Mexico in 2012, Brazil in 2013, and Uruguay in 2013, with all instances from Enterobacteriaceae.5 The blaNDM-1 gene in non-fermentative pathogens was first reported in Latin America from Honduras (Acinetobacter baumannii) and Paraguay (Acinetobacter pittii) in 2012, and later from Brazil (A. baumannii) in 2014, Cuba (Acinetobacter soli) in 2015, Argentina (Acinetobacter junii) in 2016, and Colombia (A. baumannii) in 2017.5–8 In Peru, the first report of blaNDM-1 was in May 2017 in a set of nine Klebsiella pneumoniae infecting or colonizing critically ill neurological patients from one hospital in Lima.9 Here, we describe the identification of the first three strains of A. baumannii harboring blaNDM-1 in Peru as part of a strain-based surveillance project carried out by the Naval Medical Research Unit No Six (NAMRU-6) in Lima, Peru.

Rickettsia and Leptospira spp. are under-recognized causes of acute febrile disease worldwide. Rickettsia species are often placed into the spotted fever group rickettsiae (SFGR) and typhus group rickettsiae (TGR). We explored the antibody prevalence among humans for these two groups of rickettsiae in four regions of Peru (Lima, Cusco, Puerto Maldonado, and Tumbes) and for Leptospira spp. in Puerto Maldonado and Tumbes. We also assessed risk factors for seropositivity and collected serum samples and ectoparasites from peri-domestic animals from households in sites with high human seroprevalence. In total, we tested 2,165 human sera for antibodies (IgG) against SFGR and TGR by ELISA and for antibodies against Leptospira by a microscopic agglutination test. Overall, human antibody prevalence across the four sites was 10.6% for SFGR (ranging from 6.2% to 14.0%, highest in Tumbes) and 3.3% for TGR (ranging from 2.6% to 6.4%, highest in Puerto Maldonado). Factors associated with seroreactivity against SFGR were male gender, older age, contact with backyard birds, and working in agriculture or with livestock. However, exposure to any kind of animal within the household decreased the odds ratio by half. Age was the only variable associated with higher TGR seroprevalence. The prevalence of Leptospira was 11.3% in Puerto Maldonado and 5.8% in Tumbes, with a borderline association with keeping animals in the household. We tested animal sera for Leptospira and conducted polymerase chain reaction (PCR) to detect Rickettsia species among ectoparasites collected from domestic animals in 63 households of seropositive participants and controls. We did not find any association between animal infection and human serostatus.

Leptospirosis is a neglected zoonotic disease with worldwide endemicity and continues to be a significant public health burden on resource-limited populations. Previously, we produced three highly purified recombinant antigens (rLipL32, rLipL41, and rLigA-Rep) and evaluated their performance of detecting Leptospira-specific antibodies in enzyme-linked immunosorbent assay (ELISA) as compared with the microscopic agglutination test (MAT). The overall sensitivity of this assay approached 90%. Recently, another recombinant antigen (rLigB-Rep) was prepared. We tested each individual antigen and a 1:1:1:1 mixture of these four antigens for the detection of Leptospira-specific antibodies in ELISA. The performance of these recombinant antigens was evaluated with a much larger febrile patient panel (337 MAT-confirmed positive sera and 92 MAT-negative sera from febrile patients). Combining the detection results of immunoglobulin M and immunoglobulin G from these four individual antigens, the overall sensitivity was close to 90% but the specificity was only 66%, based on the MAT reference method. The overall sensitivity and specificity of the four-antigen mixture were 82% and 86%, respectively. The mixture of four antigens also exhibited a broader reactivity with MAT-positive samples of 18 serovars from six major pathogenic Leptospira species. Given the limitations of MAT, the data were further analyzed by Bayesian latent class model, showing that ELISA using a 1:1:1:1 mixture still maintained high sensitivity (79%) and specificity (88%) as compared with the sensitivity (90%) and specificity (83%) of MAT. Therefore, ELISA using a mixture of these four antigens could be a very useful test for seroprevalence studies.

Introduction. Pseudomonas aeruginosa has the ability to acquire plasmids and other mobile genetic elements that confer resistance to antibiotics. Bacterial genes encoding different β-lactamases (bla), such as metallo-β-lactamases (MBLs) and extended-spectrum β-lactamases (ESBL), can confer resistance to multiple classes of β-lactam antibiotics. Case presentation. An 83 year old female was admitted in 2012 to the Peruvian Naval Hospital, Centro Medico Naval ‘Cirujano Mayor Santiago Tavara’ (CEMENA), in Lima, Peru. A midstream urine sample was collected and sent to the local CEMENA laboratory for routine urine culture. P. aeruginosa was isolated and initial antibiotic susceptibility testing showed it to be sensitive to imipenem. The clinicians started a course of meropenem, but the patient did not improve. After 5 days, a second urine culture was performed and a P. aeruginosa was isolated again, but this time the strain showed resistance to imipenem. The treatment course was changed to fosfomycin and the patient improved. Phenotypic and molecular laboratory testing to characterize the antibiotic resistance were performed, demonstrating the presence of both MBL and ESBL genes. Conclusion. To our knowledge, this is the first report of a P. aeruginosa XDR clinical isolate that co-expresses an MBL (VIM-2), OXA-1 beta-lactamase and the ESBL (GES-1) in Peru. It is also the first report of a VIM carbapenemase in Peru.

Microarrays are a valuable tool for analysis of both bacterial and eukaryotic nucleic acids. As many of these applications use non-specific amplification to increase sample concentration prior to analysis, the methods used to fragment and label large amplicons are important to achieve the desired analytical selectivity and specificity. Here, we used eight sequenced ESKAPE pathogens to determine the effect of two methods of whole genome amplicon fragmentation and three methods of subsequent labeling on microarray performance; nick translation was also assessed. End labeling of both initial DNase I-treated and sonication-fragmented amplicons failed to provide detectable material for a significant number of sequence-confirmed genes. However, processing of amplicons by nick translation, or by sequential fragmentation and labeling by Universal Labeling System or Klenow fragment/random primer provided good sensitivity and selectivity, with marginally better results obtained by Klenow fragment labeling. Nick-translation provided 91–100% sensitivity and 100% specificity in the tested strains, requiring half as many manipulations and less than 4h to process samples for hybridization; full sample processing from whole genome amplification to final data analysis could be performed in less than 10h. The method of template denaturation before amplification did affect detection sensitivity/selectivity of nick-labeled amplicons, however.

Background Antimicrobial resistance (AMR) is a growing public health threat around the world and is not well characterized in the developing setting. Specifically, there is a lack of information regarding nasal colonization with S. aureus and methicillin-resistant Staphylococcus aureus (MRSA) in Latin America and Peru. Methods This is the report of the baseline findings of a prospective cohort study followed up over 1 year at four geographically and ecologically distinct Peruvian Air Force bases in order to determine S. aureus nasal colonization prevalence and risk factors. Additionally, all MRSA isolates underwent molecular analysis which included pulsed-field gel electrophoresis and determination of virulence and resistance genes. Results We enrolled 756 military personnel. Anterior nares colonization with Staphylococcus aureus was detected in 73 of 756 participants (9.7 %) and MRSA was detected in 2 of 756 (0.3 %). Colonization rates differed significantly (P = 0.02) between geographic enrollment sites: Talara-4.3 %, Iquitos-9.1 %, Arequipa-14.0 % and Lima-11.3 %. Risk factors for S. aureus colonization included being male and a reported history of respiratory disease. Conclusion Overall, we found low prevalence of S. aureus and MRSA nasal colonization in this Peruvian military population. These findings contribute to the overall epidemiological understanding of S. aureus and MRSA in Latin America. The colonization rates which varied based on geographical location warrants further study.

Author(s): Cortez, Valerie; Canal, Enrique; Dupont-Turkowsky, J Catherine; Quevedo, Tatiana; Albujar, Christian; Chang, Ti-Cheng; Salmon-Mulanovich, Gabriela; Guezala-Villavicencio, Maria C; Simons, Mark P; Margolis, Elisa; Schultz-Cherry, Stacey; Pacheco, Victor; Bausch, Daniel G | Abstract: BackgroundThe southern Amazon Basin in the Madre de Dios region of Peru has undergone rapid deforestation and habitat disruption, leading to an unknown zoonotic risk to the growing communities in the area.Methodology/principal findingsWe surveyed the prevalence of rodent-borne Leptospira and Bartonella, as well as potential environmental sources of human exposure to Leptospira, in 4 communities along the Inter-Oceanic Highway in Madre de Dios. During the rainy and dry seasons of 2014-2015, we captured a total of 97 rodents representing 8 genera in areas that had experienced different degrees of habitat disturbance. Primarily by using 16S metagenomic sequencing, we found that most of the rodents (78%) tested positive for Bartonella, whereas 24% were positive for Leptospira; however, the patterns differed across seasons and the extent of habitat disruption. A high prevalence of Bartonella was identified in animals captured across both trapping seasons (72%-83%) and the relative abundance was correlated with increasing level of land disturbance. Leptospira-positive animals were more than twice as prevalent during the rainy season (37%) as during the dry season (14%). A seasonal fluctuation across the rainy, dry, and mid seasons was also apparent in environmental samples tested for Leptospira (range, 55%-89% of samples testing positive), and there was a high prevalence of this bacteria across all sites that were sampled in the communities.Conclusions/significanceThese data indicate the need for increased awareness of rodent-borne disease and the potential for environmental spread along the communities in areas undergoing significant land-use change.

We produced three highly purified recombinant antigens rLipL32, rLipL41, and rLigA-Rep (leptospiral immunoglobulin-like A repeat region) for the detection of Leptospira-specific antibodies in an enzyme-linked immunosorbent assay (ELISA). The performance of these recombinant antigens was evaluated using 121 human sera. Among them, 63 sera were microscopic agglutination test (MAT)-confirmed positive sera from febrile patients in Peru, 22 sera were indigenous MAT-negative febrile patient sera, and 36 sera were from patients with other febrile diseases from Southeast Asia, where leptospirosis is also endemic. Combining the results of immunoglobulin M (IgM) and IgG detection from these three antigens, the overall sensitivity is close to 90% based on the MAT. These results suggest that an ELISA using multiple recombinant antigens may be used as an alternative method for the detection of Leptospira-specific antibodies.

We investigated the etiology of acute diarrhea among Peruvian military recruits undergoing three months of basic combat training near the Amazonian city of Iquitos. From January through September 2002, 307 of 967 recruits were seen at the Health Post for diarrhea (attack rate [AR] = 31.8%, incidence = 1.28 95% confidence interval [CI] = 1.14-1.43] episodes/person-year). Shigella spp. were the most common bacterial pathogen recovered from recruits experiencing diarrhea episodes. These bacteria were isolated from 89 (40%) of 225 diarrheal stools examined (AR = 7.6%, incidence = 0.30 [95% CI = 0.24-0.38] episodes/person-year). Most (83 of 90; 92%) of the Shigella isolates were S. flexneri, of which 57 (69%) were serotype 2a. Seventy-six percent of Shigella isolates were resistant to sulfamethoxazole/trimethoprim and all were sensitive to ciprofloxacin. Peruvian soldiers may be an excellent population in which to test the efficacy of S. flexneri vaccines in advanced development.

While studying chronic verruga peruana infections in Peru from 2003, we isolated a novel Bartonella agent, which we propose be named Candidatus Bartonella ancashi. This case reveals the inherent weakness of relying solely on clinical syndromes for diagnosis and underscores the need for a new diagnostic paradigm in developing settings.

Background Leptospirosis is a potentially lethal zoonosis mainly affecting low-resource tropical countries, including Peru and its neighbouring countries. Timely diagnosis of leptospirosis is critical but may be challenging in the regions where it is most prevalent. The serodiagnostic gold standard microagglutination test (MAT) may be technically prohibitive. Our objective in this study was to assess the sensitivity, specificity, and predictive value of an IgM antibody capture enzyme-linked immunoassay (MAC-ELISA) derived from the M20 strain of Leptospira interrogans serovar Copenhageni (M20) by comparison to MAT, which was used as the gold standard method of diagnosis. Methods Acute and convalescent sera from participants participating in a passive febrile surveillance study in multiple regions of Peru were tested by both IgM MAC-ELISA and MAT. The sensitivity, specificity, positive and negative predictive value (PPV, NPV) of the MAC-ELISA assay for acute, convalescent and paired sera by comparison to MAT were calculated. Results The sensitivity, specificity, PPV and NPV of the MAC-ELISA assay for acute sera were 92.3%, 56.0%, 35.3% and 96.6% respectively. For convalescent sera, the sensitivity, specificity, PPV and NPV of the MAC-ELISA assay were 93.3%, 51.5%, 63.6% and 89.5% respectively. For paired sera, the sensitivity, specificity, PPV and NPV of the MAC-ELISA assay were 93.6%, 37.5%, 59.2%, 85.7% respectively. Conclusions The M20 MAC-ELISA assay performed with a high sensitivity and low specificity in the acute phase of illness. Sensitivity was similar as compared with MAT in the convalescent phase and specificity remained low. Paired sera were the most sensitive but least specific by comparison to MAT serodiagnosis. NPV for acute, convalescent and paired sera was high. The limited specificity and high sensitivity of the MAC-ELISA IgM suggests that it would be most valuable to exclude leptospirosis in low-resource regions that lack immediate access to definitive reference laboratory techniques such as MAT.

Antimicrobial resistance (AMR) is a growing public health threat around the world and is not well characterized in the developing setting. Specifically, there is a lack of information regarding nasal colonization with S. aureus and methicillin-resistant Staphylococcus aureus (MRSA) in Latin America and Peru. This is the report of the baseline findings of a prospective cohort study followed up over 1 year at four geographically and ecologically distinct Peruvian Air Force bases in order to determine S. aureus nasal colonization prevalence and risk factors. Additionally, all MRSA isolates underwent molecular analysis which included pulsed-field gel electrophoresis and determination of virulence and resistance genes. We enrolled 756 military personnel. Anterior nares colonization with Staphylococcus aureus was detected in 73 of 756 participants (9.7 %) and MRSA was detected in 2 of 756 (0.3 %). Colonization rates differed significantly (P = 0.02) between geographic enrollment sites: Talara-4.3 %, Iquitos-9.1 %, Arequipa-14.0 % and Lima-11.3 %. Risk factors for S. aureus colonization included being male and a reported history of respiratory disease. Overall, we found low prevalence of S. aureus and MRSA nasal colonization in this Peruvian military population. These findings contribute to the overall epidemiological understanding of S. aureus and MRSA in Latin America. The colonization rates which varied based on geographical location warrants further study.