CC8 MRSA Strains Harboring SCCmec Type IVc are Predominant in Colombian Hospitals

BACKGROUND: Recent reports highlight the incursion of community-associated MRSA within healthcare settings. However, knowledge of this phenomenon remains limited in Latin America. The aim of this study was to evaluate the molecular epidemiology of MRSA in three tertiary-care hospitals in Medellín, Colombia. METHODS: An observational cross-sectional study was conducted from 2008-2010. MRSA infections were classified as either community-associated (CA-MRSA) or healthcare-associated (HA-MRSA), with HA-MRSA further classified as hospital-onset (HAHO-MRSA) or community-onset (HACO-MRSA) according to standard epidemiological definitions established by the U.S. Centers for Disease Control and Prevention (CDC). Genotypic analysis included SCCmec typing, spa typing, PFGE and MLST. RESULTS: Out of 538 total MRSA isolates, 68 (12.6%) were defined as CA-MRSA, 243 (45.2%) as HACO-MRSA and 227 (42.2%) as HAHO-MRSA. The majority harbored SCCmec type IVc (306, 58.7%), followed by SCCmec type I (174, 33.4%). The prevalence of type IVc among CA-, HACO- and HAHO-MRSA isolates was 92.4%, 65.1% and 43.6%, respectively. From 2008 to 2010, the prevalence of type IVc-bearing strains increased significantly, from 50.0% to 68.2% (p = 0.004). Strains harboring SCCmec IVc were mainly associated with spa types t1610, t008 and t024 (MLST clonal complex 8), while PFGE confirmed that the t008 and t1610 strains were closely related to the USA300-0114 CA-MRSA clone. Notably, strains belonging to these three spa types exhibited high levels of tetracycline resistance (45.9%). CONCLUSION: CC8 MRSA strains harboring SCCmec type IVc are becoming predominant in Medellín hospitals, displacing previously reported CC5 HA-MRSA clones. Based on shared characteristics including SCCmec IVc, absence of the ACME element and tetracycline resistance, the USA300-related isolates in this study are most likely related to USA300-LV, the recently-described 'Latin American variant' of USA300

In Vitro and In Vivo Efficacy of Ether Lipid Edelfosine against Leishmania spp. and SbV-Resistant Parasites

Leishmaniasis represents a major international health problem, has a high morbidity and mortality rate, and is classified as an emerging and uncontrolled disease by the World Health Organization. The migration of population from endemic to nonendemic areas, and tourist activities in endemic regions are spreading the disease to new areas. Unfortunately, treatment of leishmaniasis is far from satisfactory, with only a few drugs available that show significant side-effects. Here, we show in vitro and in vivo evidence for the antileishmanial activity of the ether phospholipid edelfosine, being effective against a wide number of Leishmania spp. causing cutaneous, mucocutaneous and visceral leishmaniasis. Our experimental mouse and hamster models demonstrated not only a significant antileishmanial activity of edelfosine oral administration against different wild-type Leishmania spp., but also against parasites resistant to pentavalent antimonials, which constitute the first line of treatment worldwide. In addition, edelfosine exerted a higher antileishmanial activity and a lower proneness to generate drug resistance than miltefosine, the first drug against leishmaniasis that can be administered orally. These data, together with our previous findings, showing an anti-inflammatory action and a very low toxicity profile, suggest that edelfosine is a promising orally administered drug for leishmaniasis, thus warranting clinical evaluation

Eco-epidemiological analysis of rickettsial seropositivity in rural areas of Colombia: A multilevel approach

ABSTARCT: Rickettsiosis is a re-emergent infectious disease without epidemiological surveillance in Colombia. This disease is generally undiagnosed and several deadly outbreaks have been reported in the country in the last decade. The aim of this study is to analyze the eco-epidemiological aspects of rickettsial seropositivity in rural areas of Colombia where outbreaks of the disease were previously reported. A cross-sectional study, which included 597 people living in 246 households from nine hamlets in two municipalities of Colombia, was conducted from November 2015 to January 2016. The survey was conducted to collect sociodemographic and household characteristics (exposure) data. Blood samples were collected to determine the rickettsial seropositivity in humans, horses and dogs (IFA, cut-off = 1/128). In addition, infections by rickettsiae were detected in ticks from humans and animals by real-time PCR targeting gltA and ompA genes. Data was analyzed by weighted multilevel clog-log regression model using three levels (person, household and hamlets) and rickettsial seropositivity in humans was the main outcome. Overall prevalence of rickettsial seropositivity in humans was 25.62% (95%CI 22.11-29.12). Age in years (PR = 1.01 95%CI 1.01-1.02) and male sex (PR = 1.65 95%CI 1.43-1.90) were risk markers for rickettsial seropositivity. Working outdoors (PR = 1.20 95%CI 1.02-1.41), deforestation and forest fragmentation for agriculture use (PR = 1.75 95%CI 1.51-2.02), opossum in peridomiciliary area (PR = 1.56 95%CI 1.37-1.79) and a high proportion of seropositive domestic animals in the home (PR20-40% vs 40% vs <20% = 3.14 95%CI 2.43-4.04) were associated with rickettsial seropositivity in humans. This study showed the presence of Rickettsia antibodies in human populations and domestic animals. In addition, different species of rickettsiae were detected in ticks collected from humans and animals. Our results highlighted the role of domestic animals as sentinels of rickettsial infection to identify areas at risk of transmission, and the importance of preventive measures aimed at curtailing deforestation and the fragmentation of forests as a way of reducing the risk of transmission of emergent and re-emergent pathogens

Detección de Brucella abortus por PCR en sangre y leche de vacunos

Objetivo. Evaluar el uso de la Reacción en Cadena de la Polimerasa (PCR) para la detección de Brucella abortus en muestras de sangre y leche de vacunos. Materiales y métodos. Este estudio de tipo descriptivo fue realizado durante los años 2004 y 2005. Se analizaron 136 animales de tres fincas localizadas en el municipio de Durania, Norte de Santander, Colombia. Se evaluó la presencia de anticuerpos en la leche mediante la prueba del anillo (PAL). Se amplifico el fragmento de 223pb del gen BCSP31. Se emplearon los cebadores B4 y B5 de la región interna de la secuencia del gen BCSP31 (GenBank, número de M20404). Resultados. En aquellos animales positivos se obtuvo una muestra de sangre y leche para el análisis por PCR, la sangre no fue analizada por serología. Se evaluaron diferentes métodos de extracción de ADN. Se encontró que un 13.2% (18/136) de las muestras de leche fueron positivas a la PAL. Se analizaron 33 muestras de leche negativas por PAL de las cuales el 30.3% (10/33) resultaron positivas por PCR. Al analizar las muestras de sangre de los animales positivos por PAL el 94.1% (16/17) fueron positivas por PCR, mientras que el 47% (8/17) de las muestras de leche positivas por PAL, fueron positivas por PCR. Conclusiones. Se demostró la amplificación de un fragmento de ADN de Brucella abortus en muestras de sangre y leche de vacunos. Los resultados preliminares demuestran que es posible usar PCR como prueba diagnóstica de brucelosis en Colombia

Peri-articular injection of an analgesic mixture in primary total hip arthroplasty: an effective strategy for pain control during the first post-operative day

Background: Previous studies of soft tissue infiltration in hip arthroplasty present variable results. The purpose of this study is to identify whether injection of an analgesic mixture improves pain management during the immediate post-operative period. Materials and methods: This cohort study compared 129 patients that received peri-articular soft tissue injection with 20 ml of 0.25% bupivacaine and 2 ml of ketorolac (30 mg/1 ml) in 28 ml of saline solution, with 71 patients who did not received injections. Pain intensity in the Verbal Analog Scale (VAS), opioid titration, and consumption (mg morphine equivalents) in the post-anaesthetic care unit (PACU) and during the first post-operative day were assessed for both groups. All patients received the same analgesia protocol. Results: Median VAS score in the PACU was 4 (IQR 2–7) in the injection group and 7 (IQR 4–8) in the non-injection group (p = 0.001). Median opioid titration was 0 mg for the injection group and 2.6 mg for the non-injection group (p = 0.011). In the first post-operative day, the difference in VAS scores between groups was statistically significant (p = 0.009), but there was no difference in opioid consumption. Conclusion: Soft tissue injection with local anesthetics and non-steroidal anti-inflammatory drugs allows adequate pain control in the immediate post-operative period and reduces the requirement for opioid consumption. We recommend the implementation of this safe and effective strategy in post-operative pain management after primary hip arthroplasty. Level of evidence: Level II, cohort study. © 2018, SICOT aisbl

Detection in silico of second use drugs with potential anti-leishmania activity

Actualmente las enfermedades tropicales son objeto de importantes investigaciones en las ciencias biomédicas, debido al impacto global que causa en poblaciones vulnerables. Sin embargo son comúnmente ignoradas por la industria farmacéutica debido a su bajo potencial de rentabilidad económica. Por esta razón, la búsqueda de nuevos tratamientos terapéuticos por medios costo-efectivos es esencial para la lucha contra parásitos tropicales como Leishmania spp. En este trabajo se hizo una búsqueda y selección de medicamentos depositados en bases de datos públicas, y cuyo blanco de acción demostrado son proteínas conocidas. Con estas proteínas blanco de medicamentos, se hizo una búsqueda de ortólogos en el proteoma de Leishmania, con el fi n de identifi car rápidamente medicamentos que pudieran tener acción también contra este parásito, implementando herramientas in silico, basadas en la Bioinformática. También en el caso de poseer la estructura de las proteínas de interés, se realizó análisis de docking para corroborar la interacción con el medicamento. Empleando esta estrategia, se identifi caron y seleccionaron 10 medicamentos que son evaluados actualmente en ensayos in vitro.Currently, tropical diseases are a major subject of research in biomedical sciences due to its global impact on vulnerable populations. However, these diseases are normally ignored by pharmaceutical companies due to low profi tability potential. For that reason, the search of new therapeutic treatments, that are cost-effective, is essential to fi ght against tropical parasites as Leishmania spp. In this work, a search and selection of drugs with known protein targets, which were deposited in public databases, was conducted. With these target proteins, a search for orthologs in the Leishmania proteome was carried out in order to identify drugs that could also have anti-leishmanial activity. For this purpose bioinformatics tools were implemented. In addition, in the case that Leishmania proteins have its tridimensional structure reported, docking analysis were simulated to corroborate interaction with the drug. At the end, a selection of 10 drugs was identifi ed and is currently being evaluated in in vitro tes

Differentiation of Leishmania (Viannia) panamensis and Leishmania (V.) guyanensis using BccI for hsp70 PCR-RFLP

Leishmania panamensis and Leishmania guyanensis are two species of the subgenus Viannia that are genetically very similar. Both parasites are usually associated with cutaneous leishmaniasis, but also have the potential to cause the mucocutaneous form of the disease. In addition, the study of foci and consequently the identification of vectors and probable reservoirs involved in transmission require a correct differentiation between both species, which is important at epidemiological level. We explored the possibility of identifying these species by using restriction fragment length polymorphisms (RFLP) in the gene coding for heat-shock protein 70 (hsp70). Previously, an hsp70 PCR-RFLP assay proved to be very effective in differentiating other Leishmania species when HaeIII is used as restriction enzyme. Based on hsp70 sequences analysis, BccI was found to generate species-specific fragments that can easily be recognized by agarose gel electrophoresis. Using the analysis of biopsies, scrapings, and parasite isolates previously grouped in a cluster comprising both L. panamensis and L. guyanensis, we showed that our approach allowed differentiation of both entities. This offers the possibility not only for identification of parasites in biological samples, but also to apply molecular epidemiology in certain countries of the New World, where several Leishmania species could coexist. Copyright 2009 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved

In Vitro and In Vivo Efficacy of Ether Lipid Edelfosine against Leischmania spp. and SbV-Resistant Parasites

BACKGROUND: The leishmaniases are a complex of neglected tropical diseases caused by more than 20 Leishmania parasite species, for which available therapeutic arsenal is scarce and unsatisfactory. Pentavalent antimonials (SbV) are currently the first-line pharmacologic therapy for leishmaniasis worldwide, but resistance to these compounds is increasingly reported. Alkyl-lysophospoholipid analogs (ALPs) constitute a family of compounds with antileishmanial activity, and one of its members, miltefosine, has been approved as the first oral treatment for visceral and cutaneous leishmaniasis. However, its clinical use can be challenged by less impressive efficiency in patients infected with some Leishmania species, including L. braziliensis and L. mexicana, and by proneness to develop drug resistance in vitro. METHODOLOGY/PRINCIPAL FINDINGS: We found that ALPs ranked edelfosine>perifosine>miltefosine>erucylphosphocholine for their antileishmanial activity and capacity to promote apoptosis-like parasitic cell death in promastigote and amastigote forms of distinct Leishmania spp., as assessed by proliferation and flow cytometry assays. Effective antileishmanial ALP concentrations were dependent on both the parasite species and their development stage. Edelfosine accumulated in and killed intracellular Leishmania parasites within macrophages. In vivo antileishmanial activity was demonstrated following oral treatment with edelfosine of mice and hamsters infected with L. major, L. panamensis or L. braziliensis, without any significant side-effect. Edelfosine also killed SbV-resistant Leishmania parasites in in vitro and in vivo assays, and required longer incubation times than miltefosine to generate drug resistance. CONCLUSIONS/SIGNIFICANCE: Our data reveal that edelfosine is the most potent ALP in killing different Leishmania spp., and it is less prone to lead to drug resistance development than miltefosine. Edelfosine is effective in killing Leishmania in culture and within macrophages, as well as in animal models infected with different Leishmania spp. and SbV-resistant parasites. Our results indicate that edelfosine is a promising orally administered antileishmanial drug for clinical evaluation

Rickettsial infection in ticks collected from humans and domestic animals.

<p>Rickettsial infection in ticks collected from humans and domestic animals.</p