Genomic Characterization of Extended-Spectrum Cephalosporin-Resistant Salmonella enterica in the Colombian Poultry Chain.

Salmonella enterica serovars have been isolated from Colombian broilers and broiler meat. The aim of this study was to investigate the diversity of ESBL/pAmpC genes in extended-spectrum cephalosporin resistant Salmonella enterica and the phylogeny of ESBL/pAmpC-carrying Salmonella using Whole Genome Sequencing (WGS). A total of 260 cefotaxime resistant Salmonella isolates, obtained between 2008 and 2013 from broiler farms, slaughterhouses and retail, were included. Isolates were screened by PCR for ESBL/pAmpC genes. Gene and plasmid subtyping and strain Multi Locus Sequence Typing was performed in silico for a selection of fully sequenced isolates. Core-genome-based analyses were performed per ST encountered. bla CMY-2-like was carried in 168 isolates, 52 carried bla CTX-M-2 group, 7 bla SHV, 5 a combination of bla CMY-2-like-bla SHV and 3 a combination of bla CMY-2-like-bla CTX-M-2 group. In 25 isolates no ESBL/pAmpC genes that were screened for were found. WGS characterization of 36 selected strains showed plasmid-encoded bla CMY-2 in 21, bla CTX-M-165 in 11 and bla SHV-12 in 7 strains. These genes were mostly carried on IncI1/ST12, IncQ1, and IncI1/ST231 plasmids, respectively. Finally, 17 strains belonged to S. Heidelberg ST15, 16 to S. Paratyphi B variant Java ST28, 1 to S. Enteritidis ST11, 1 to S. Kentucky ST152 and 1 to S. Albany ST292. Phylogenetic comparisons with publicly available genomes showed separate clustering of Colombian S. Heidelberg and S. Paratyphi B var. Java. In conclusion, resistance to extended-spectrum cephalosporins in Salmonella from Colombian poultry is mainly encoded by bla CMY-2 and bla CTX-M-165 genes. These genes are mostly associated with IncI1/ST12 and IncQ1 plasmids, respectively. Evolutionary divergence is observed between Colombian S. Heidelberg and S. Paratyphi B var. Java and those from other countries.

High Heterogeneity of Escherichia coli Sequence Types Harbouring ESBL/AmpC Genes on IncI1 Plasmids in the Colombian Poultry Chain.

BACKGROUND: Escherichia coli producing ESBL/AmpC enzymes are unwanted in animal production chains as they may pose a risk to human and animal health. Molecular characterization of plasmids and strains carrying genes that encode these enzymes is essential to understand their local and global spread. OBJECTIVES: To investigate the diversity of genes, plasmids and strains in ESBL/AmpC-producing E. coli from the Colombian poultry chain isolated within the Colombian Integrated Program for Antimicrobial Resistance Surveillance (Coipars). METHODS: A total of 541 non-clinical E. coli strains from epidemiologically independent samples and randomly isolated between 2008 and 2013 within the Coipars program were tested for antimicrobial susceptibility. Poultry isolates resistant to cefotaxime (MIC ≥ 4 mg/L) were screened for ESBL/AmpC genes including blaCTX-M, blaSHV, blaTEM, blaCMY and blaOXA. Plasmid and strain characterization was performed for a selection of the ESBL/AmpC-producing isolates. Plasmids were purified and transformed into E. coli DH10B cells or transferred by conjugation to E. coli W3110. When applicable, PCR Based Replicon Typing (PBRT), plasmid Multi Locus Sequence Typing (pMLST), plasmid Double Locus Sequence Typing (pDLST) and/or plasmid Replicon Sequence Typing (pRST) was performed on resulting transformants and conjugants. Multi Locus Sequence Typing (MLST) was used for strain characterization. RESULTS: In total, 132 of 541 isolates were resistant to cefotaxime and 122 were found to carry ESBL/AmpC genes. Ninety-two harboured blaCMY-2 (75%), fourteen blaSHV-12 (11%), three blaSHV-5 (2%), five blaCTX-M-2 (4%), one blaCTX-M-15 (1%), one blaCTX-M-8 (1%), four a combination of blaCMY-2 and blaSHV-12 (4%) and two a combination of blaCMY-2 and blaSHV-5 (2%). A selection of 39 ESBL/AmpC-producing isolates was characterized at the plasmid and strain level. ESBL/AmpC genes from 36 isolates were transferable by transformation or conjugation of which 22 were located on IncI1 plasmids. These IncI1 plasmids harboured predominantly blaCMY-2 (16/22), and to a lesser extend blaSHV-12 (5/22) and blaCTX-M-8 (1/22). Other plasmid families associated with ESBL/AmpC-genes were IncK (4/33), IncHI2 (3/33), IncA/C (2/33), IncΒ/O (1/33) and a non-typeable replicon (1/33). Subtyping of IncI1 and IncHI2 demonstrated IncI1/ST12 was predominantly associated with blaCMY-2 (12/16) and IncHI2/ST7 with blaCTX-M-2 (2/3). Finally, 31 different STs were detected among the 39 selected isolates. CONCLUSIONS: Resistance to extended spectrum cephalosporins in E. coli from Colombian poultry is mainly caused by blaCMY-2 and blaSHV-12. The high diversity of strain Sequence Types and the dissemination of homogeneous IncI1/ST12 plasmids suggest that spread of the resistance is mainly mediated by horizontal gene transfer.

Whole-Genome Sequences of Two Campylobacter coli Isolates from the Antimicrobial Resistance Monitoring Program in Colombia.

Campylobacter coli, along with Campylobacter jejuni, is a major agent of gastroenteritis and acute enterocolitis in humans. We report the whole-genome sequences of two multidrug-resistance C. coli strains, isolated from the Colombian poultry chain. The isolates contain a variety of antimicrobial resistance genes for aminoglycosides, lincosamides, fluoroquinolones, and tetracycline.

Differences on Primary Care Labor Perceptions in Medical Students from 11 Latin American Countries.

BACKGROUND: The shortage in Latin-American Primary Care (PC) workforce may be due to negative perceptions about it. These perceptions might be probably influenced by particular features of health systems and academic environments, thus varying between countries. METHODS: Observational, analytic and cross-sectional multicountry study that evaluated 9,561 first and fifth-year medical students from 63 medical schools of 11 Latin American countries through a survey. Perceptions on PC work was evaluated through a previously validated scale. Tertiles of the scores were created in order to compare the different countries. Crude and adjusted prevalence ratios were calculated using simple and multiple Poisson regression with robust variance. RESULTS: Approximately 53% of subjects were female; mean age was 20.4±2.9 years; 35.5% were fifth-year students. Statistically significant differences were found between the study subjects' country, using Peru as reference. Students from Chile, Colombia, Mexico and Paraguay perceived PC work more positively, while those from Ecuador showed a less favorable position. No differences were found among perceptions of Bolivian, Salvadoran, Honduran and Venezuelan students when compared to their Peruvian peers. CONCLUSIONS: Perceptions of PC among medical students from Latin America vary according to country. Considering such differences can be of major importance for potential local specific interventions.

Prevalence, resistance patterns, and risk factors for antimicrobial resistance in bacteria from retail chicken meat in Colombia.

As a step toward implementing the Colombian Integrated Program for Antimicrobial Resistance Surveillance (COIPARS), this study aimed to establish the baseline antimicrobial resistance patterns of Salmonella serovars, Escherichia coli, and Enterococcus spp. isolates in retail poultry meat from independent stores and from a main chain distributor center. MICs of the isolates were determined for antimicrobials used both in humans and animals, using an automated system. Salmonella serovars were isolated from 26% of the meat samples and E. coli from 83%, whereas Enterococcus faecalis and Enterococcus faecium were detected in 81 and 13% of the meat samples, respectively. A principal finding of concern in this study was that almost 98% of isolates tested were multidrug resistant. Ceftiofur, enrofloxacin, nalidixic acid, and tetracycline were the antimicrobials that showed the highest frequency of resistance among Salmonella and E. coli isolates. For enterococci, 61.5% of E. faecium isolates were found to be resistant to quinupristin-dalfopristin; this is significant because it is used to treat nosocomial infections when vancomycin resistance is present. Vancomycin resistance was detected in 4% of the E. faecalis isolates. The results of our study highlight the need for rapid implementation of an integrated program for surveillance of antimicrobial resistance by the Colombian authorities in order to monitor trends, raise awareness, and help promote practices to safeguard later generation antimicrobial agents.

Molecular characterization of Salmonella paratyphi B dT+ and Salmonella Heidelberg from poultry and retail chicken meat in Colombia by pulsed-field gel electrophoresis.

Salmonella Paratyphi B dT+ variant (also termed Salmonella Java) and Salmonella Heidelberg are pathogens of public health importance that are frequently isolated from poultry. As a step toward implementing the Colombian Integrated Program for Antimicrobial Resistant Surveillance, this study characterized molecular patterns of Salmonella Paratyphi B dT+ and Salmonella Heidelberg isolated from poultry farms, fecal samples, and retail chicken meat using pulsed-field gel electrophoresis (PFGE). The objective of this study was to determine the genetic relationship among isolates and to determine potential geographically predominant genotypes. Based on PFGE analysis, both serovars exhibited high heterogeneity: the chromosomal DNA fingerprints of 82 Salmonella Paratyphi B dT+ isolates revealed 42 PFGE patterns, whereas the 21 isolates of Salmonella Heidelberg revealed 10 patterns. Similar genotypes of both serovars were demonstrated to be present on farms and in retail outlets. For Salmonella Paratyphi B dT+, closely genetically related strains were found among isolates coming from different farms and different integrated poultry companies within two departments (Santander and Cundinamarca) and also from farms located in the two geographically distant departments. For Salmonella Heidelberg, there were fewer farms with genetically related isolates than for Salmonella Paratyphi B dT+. A possible dissemination of similar genotypes of both serovars along the poultry production chain is hypothesized, and some facilitating factors existing in Colombia are reviewed.

Counts, serovars, and antimicrobial resistance phenotypes of Salmonella on raw chicken meat at retail in Colombia.

The objective of this study was to determine Salmonella counts, serovars, and antimicrobial-resistant phenotypes on retail raw chicken carcasses in Colombia. A total of 301 chicken carcasses were collected from six departments (one city per department) in Colombia. Samples were analyzed for Salmonella counts using the most-probable-number method as recommended by the U.S. Department of Agriculture, Food Safety Inspection Service protocol. A total of 378 isolates (268 from our previous study) were serotyped and tested for antimicrobial susceptibility. The overall Salmonella count (mean log most probable number per carcass ± 95% confidence interval) and prevalence were 2.1 (2.0 to 2.3) and 37%, respectively. There were significant differences (P < 0.05) by Salmonella levels (i.e., counts and prevalence) by storage temperature (i.e., frozen, chilled, or ambient), retail store type (wet markets, supermarkets, and independent markets), and poultry company (chicken produced by integrated or nonintegrated company). Frozen chicken had the lowest Salmonella levels compared with chicken stored at other temperatures, chickens from wet markets had higher levels than those from other retail store types, and chicken produced by integrated companies had lower levels than nonintegrated companies. Thirty-one Salmonella serovars were identified among 378 isolates, with Salmonella Paratyphi B tartrate-positive (i.e., Salmonella Paratyphi B dT+) the most prevalent (44.7%), followed by Heidelberg (19%), Enteritidis (17.7%), Typhimurium (5.3%), and Anatum (2.1%). Of all the Salmonella isolates, 35.2% were resistant to 1 to 5 antimicrobial agents, 24.6% to 6 to 10, and 33.9% to 11 to 15. Among all the serovars obtained, Salmonella Paratyphi B dT+ and Salmonella Heidelberg were the most antimicrobial resistant. Salmonella prevalence was determined to be high, whereas cell numbers were relatively low. These data can be used in developing risk assessment models for preventing the transmission of Salmonella from chicken to humans in Colombia.

Prevalence of Salmonella on retail broiler chicken meat carcasses in Colombia.

A cross-sectional study was performed to estimate the prevalence of Salmonella on retail market chicken carcasses in Colombia. A total of 1,003 broiler chicken carcasses from 23 departments (one city per department) were collected via a stratified sampling method. Carcass rinses were tested for the presence of Salmonella by conventional culture methods. Salmonella strains were isolated from 27 % of the carcasses sampled. Logistic regression analysis was used to determine potential risk factors for Salmonella contamination associated with the chicken production system (conventional versus free-range), storage condition (chilled versus frozen), retail store type (supermarket, independent, and wet market), poultry company (integrated company versus nonintegrated company), and socioeconomic stratum. Chickens from a nonintegrated poultry company were associated with a significantly (P < 0.05) greater risk of Salmonella contamination (odds ratio, 2.0) than were chickens from an integrated company. Chilled chickens had a significantly (P < 0.05) higher risk of Salmonella contamination (odds ratio, 4.3) than did frozen chicken carcasses.

A 25-Year Review of Cuba's Screening Program for Early Detection of Hearing Loss.

Introduction Early screening for hearing loss is currently recognized as an international healthcare standard. In Cuba, such a program was initiated in the capital, Havana, in 1983 and scaled up to national coverage in 1991. Objective Review the development of Cuba's national hearing screening program over the last 25 years (organizational structure, efficiency, coverage and impact on health), and the science and technology developed to sustain it. Intervention The program was organized in two steps: Step 1-clinical selection of children at different stages of development with multiple high-risk registers; Step 2-referral to territorial, hospitalbased centers for auditory brainstem evoked response (ABR) testing, diagnostic evaluation, and intervention. Prior to national scaling-up, the efficiency of this multiple targeted screening (MTS) protocol was evaluated in Havana. Technology and equipment were then developed, and personnel were trained to set up the national screening network. In 1996, the multiple auditory steady-state evoked response (MSSR) technique for objective audiogram estimation was introduced using AUDIX equipment, designed and produced in Cuba for this program. A semi-automated version for neonatal screening has been developed more recently. Several studies have been conducted to evaluate the program's efficiency, coverage, yield, and impact on health. Results During the first stage of implementation in Havana, the MTS protocol correctly identified 72.5% of children with congenital and preverbal hearing loss. Subsequent studies of different aspects of the program have shown that: 1) the mean age of hearing loss detection/intervention in one municipality was reduced from 4 years to 10 months; 2) hearing-impaired children who were screened showed improved language and cognitive development compared to those who were not screened; 3) the MSSR technique predicted type and severity of hearing loss more accurately than physiological techniques used previously and was also shown to be an effective screening method (92% to 96% sensitivity, 100% specificity); and 4) program coverage (25-86%), though reasonably high in some regions, is not complete and needs improvement, particularly in the country's remote and rural areas. Conclusions The MTS protocol can be considered a valid option for increasing the yield and effectiveness of a hearing screening program operating with limited resources. The MSSR technique provides valuable data for the diagnosis and treatment of children detected through a screening program and, with improvements, may also be useful as a screening method.

Sorderas neurosensoriales no sindrómicas: análisis de la herencia en 10 familias / Non syndromic neurosensorial deafness: analysis of the hereditary in 20 families

Se reportan los árboles genealógicos de 10 probandos afectados con sorderas neurosensoriales no sindrómicas de aparición familiar. El análisis genético practicado permitió reconocer la clara segregación de un único gen de sordera en 7 familias (3 autosómicas recesivas, 2 autosómicas dominantes, 1 ligada al cromosoma X, 1 con herencia mitocondrial). En las 3 familias restantes resultó difícil el análisis y se propuso la herencia recesiva como la más probable, sobre la base, fundamentalmente, de las características de la pérdida auditiva (congénita, bilateral, severa o profunda). En general las sorderas autosómicas recesivas fueron las más frecuentes. Se corrobora que estos estudios suelen ser complicados por la gran heterogeneidad que pueden presentar a todos los niveles las sorderas neurosensoriales no sindrómicas

Mutaciones del gen de la conexina 26 (GJB2) en familias cubanas con sorderas no sindrómicas autosómicas recesivas / Mutations in the connexion 26 gen (GJB2) among cuban families with non-syndromic autosomal recessive deafness

Las mutaciones del gen de la conexina 26 (locus DFNB1, en el brazo largo del cromosoma 13) dan cuenta de 60 porciento de las familias con sorderas neurosensoriales no sindrómicas autosómicas recesivas en poblaciones caucásicas. La prueba para la detección de la mutación 35delG, el análisis de heteroduplex y la secuenciación de la región codificante del gen de la conexina 26 en miembros de 15 familias cubanas con este tipo de sordera, arrojaron los resultados siguientes: en 10 de 15 familias (66,66 porciento) se observaron mutaciones en ambos alelos de la conexina 26, por lo que se clasificaron como sorderas tipo DFNB1. en estas familias DFNB1, 25/32 cromosomas analizados contenían la mutación 35deIG para una frecuencia de 0,781. Las mutaciones E47X y W77R se observaron en heterocigosis con la 35deIG. En 2 hipoacúsicos se detectaron también en heterocigosis las mutaciones puntuales M34T/R143W y V95M/R184P, respectivamente. Los hallazgos permitieron apreciar la alta frecuencia del tipo de sordera DFNB1 entre las sorderas no sindrómicas autosómicas recesivas que existían en la población cubana. El aspecto más destacable de la investigación es la similitud de sus resultados con los encontrados en poblaciones caucásicas relacionadas con el origen étnico de la población cubana. Este trabajo constituye la primera comunicación de estos estudios en el medio cubano

Expresión fenotípica de una sordera familiar con deleción del gen POU3F4 / Phenotype expression of a familial deafness with deletion of POU3F4 gene

Se presenta una familia cubana con 5 miembros afectados por una hipoacusia bilateral, congénita, severa, mixta con componente neurosensorial predominante y sin alteraciones morfológicas de oído interno. El patrón de transmisión era compatible con la herencia recesiva ligada al cromosoma X. Los estudios moleculares detectaron una deleción en la región Xq21.1 que implica el gen POU3F4, responsable de la sordera de tipo DFN3. Se hacen comentarios sobre la evidente variabilidad clínica de las sorderas tipo DFN3

Interferón en papilomatosis laringea infantil / Interferon in infantile laryngeal papillomatosis

Se conoce que la papilomatosis laríngea es una enfermedad de origen viral, para la cual no ha existido un tratamiento médico específico. También se conoce que el interferón leucocitario humano es un agente antiviral específico que inhibe la replicación viral intracelular. En diferentes estudios se ha utilizado el interferón y se muestran resultados favorables con esta terapéutica. Persisten interrogantes en cuanto a los esquemas y dosis para controlar definitivamente la enfermedad. Se trataron 5 pacientes que presentaban papilomatosis laríngea, con interferón leucocitario humano por vía intramuscular en dosis crecientes. Los pacientes fueron examinados clínicamente todos los meses. Los 5 niños en tratamiento han evolucionado favorablemente. Sólo en 1 caso hubo proliferación de tejido papilomatoso. La voz y función respiratoria han mejorado ostensiblemente en el curso del tratamiento