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Integrons are prokaryotic genetic elements known to carry and exchange antibiotic resistance gene cassettes through a site-specific recombinase called integrase. In this work, 107 Aeromonas isolates from environmental origin, including fish, water, and sediments, were investigated for the presence of integrons. Using specific primers for Class 1, 2 and 3 integrases, only Class 1 and Class 2 integrons were detected. Detection of Class 2 integrases and their associated variable regions required two rounds of polymerase chain reaction (PCR). Sequencing of the intI2 amplicons confirmed them as integrase-derived products. Class 1 integrons were detected in 26 out of 107 isolates. PCR amplification of the variable regions associated to these integrons revealed an outstanding homogeneity, 25 of them having variable regions with an identical dfrA12-orfF-aadA2 cassette array and one integron carrying only the dfrA16 cassette. To assess clone diversity, chromosomal DNA from isolates was subjected to enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR), which discarded clonality in all instances. Class 2 integrons were surprisingly more prevalent than Class1 integrons, being detected in 60 out of 107 isolates. Forty-six of them showed a unique ERIC profile, while the remaining 14 strains displayed profiles that could be grouped in five different patterns. Cassette arrangements of all Class 2 variable regions were those described as the most prevalent (dfrA1-sat2-aadA1). A rather startling result of this work is the sensitivity to trimethoprim, streptomycin, and streptothricin of most strains, despite the presence of the cognate resistance genes. To know the integron distribution in environmental Aeromonas species, a phylogenetic reconstruction was done using rpoD/gyrB or rpoD/gyrA gene sequences. Isolates bearing these elements corresponded to Aeromonas hydrophila, Aeromonas veronii, Aeromonas salmonicida, Aeromonas dhakensis, Aeromonas sanarellii, Aeromonas taiwanensis, Aeromonas media, Aeromonas caviae, Aeromonas jandaei, and Aeromonas sp. This work revealed an unusual high incidence of Class 2 integrons and a low variability of cassette arrangements in environmental Aeromonas species.

As dendritic cells (DCs) are among the first cells to encounter antigens, these cells trigger both innate and T cell responses, and are the most potent antigen-presenting cells. Brucella spp., which is an intracellular facultative and stealthy pathogen, is able to evade the bactericidal activities of professional phagocytes. Several studies have demonstrated that Brucella can survive and replicate intracellularly, thereby provoking impaired maturation of DCs. Therefore, the interaction between DCs and Brucella becomes an interesting model to study the immune response. In this review, we first will describe the most common techniques for DCs differentiation in vitro as well as general features of brucellosis. Then, the interaction of DCs and Brucella, including pathogen recognition, molecular mechanisms of bacterial pathogenesis, and intracellular trafficking of Brucella to subvert innate response, will be reviewed. Finally, we will debate diversity in immunological DC response and the controversial role of DC activation against Brucella infection.

El agua es el vehículo por el cual se transportan diversos tipos de desechos provenientes de diferentes actividades productivas, de tal forma que ríos, canales, lagunas y mares se contaminan con éstos. Esta situación se ha presentado en México desde hace más de 200 años, y en la actualidad se están generando nuevos contaminantes no reglamentados en las aguas de reúso o tratadas. En el caso de residuos de fármacos, su persistencia y entrada continua a mantos acuíferos originan efectos negativos sobre la vida y el ambiente acuático, debido a que no se eliminan totalmente en los procesos de tratamiento de aguas residuales a causa de su estabilidad química. Este tipo de agua se reutiliza para la producción de agua potable o el riego de zonas agrícolas, de manera que la salud humana y animal se encuentran expuestas a los efectos toxicológicos de este tipo de sustancias químicas. Se realizó la búsqueda de reportes acerca de la contaminación de residuos farmacológicos en diferentes tipos de agua —residual o no— y suelos agrícolas, tanto internacionales como nacionales, de 1984 a 2018. Se concluyó que no hay hasta hoy una normatividad nacional ni internacional idónea sobre la forma en que debe realizarse la eliminación de este tipo de desechos para al medio ambiente en la menor medida posible.

Outer membrane vesicles (OMVs) from Gram-negative bacteria were first described more than 50 years ago. However, the molecular mechanisms involved in biogenesis began to be studied only in the last few decades. Presently, the biogenesis and molecular mechanisms for their release are not completely known. This review covers the most recent information on cellular components involved in OMV biogenesis, such as lipoproteins and outer membrane proteins, lipopolysaccharide, phospholipids, quorum-sensing molecules, and flagella. CONACYTConsejo Nacional de Ciencia y Tecnologia (CONACyT) [61529]; SIP-IPN [20182152, 20195737, 20200594]; SAGARPA-CONACYTConsejo Nacional de Ciencia y Tecnologia (CONACyT) [2017-02-291311]; PIFI-IPN scholarships; COFAA-IPN; SIP-EDI; SNI-CONACYTConsejo Nacional de Ciencia y Tecnologia (CONACyT) This work was funded by CONACYT 61529, SIP-IPN 20182152, 20195737, 20200594, and SAGARPA-CONACYT 2017-02-291311. EA-C was supported by CONACYT and PIFI-IPN scholarships. MA-A, ER, ZG-L, and AC-R were supported by fellowships from COFAA-IPN, SIP-EDI, and SNI-CONACYT.

The Rio Bravo (Rio Grande) adjoins various states in the Mexican region and has a great importance in water distribution in the northeast Tamaulipas (Mexico). In this work 161 strains were isolated, identified and characterized from the water samples taken from the flow of the Rio Bravo and the two inner canals that cover Reynosa city. The strains were identified as Vibrio cholerae (74·5%), Vibrio spp. (1·2%) and Vibrio mimicus (0·6%). Furthermore, the detected virulence genes in the V. cholerae strains, were the hlyA, ompU, tcpA, toxR genes in 78·3, 62·5, 15·8 and 90·8% respectively. Only the ompU and vmh genes were detected in the V. mimicus strain. These results indicate the presence of multi-toxigenic V. cholerae strains in the Rio Bravo/Grande and in the water bodies from Reynosa city, which could represent a risk for the exposed population. Significance and impact of the study Water quality is associated with public health, as it plays an important role in the transmission and epidemiology of pathogens such as Vibrio, since this species have been responsible for human diseases around the world. This study demonstrated the presence of toxigenic Vibrio species in water bodies in Reynosa surroundings, indicating that water bodies may be a source of public health risk.

Integrons are prokaryotic genetic elements known to carry and exchange antibiotic resistance gene cassettes through a site-specific recombinase called integrase. In this work, 107

The Embp protein of Staphylococcus epidermidis inhibits the hemagglutination of the H1N1 influenza virus and protects birds from a viral respiratory infection. Several species of Coagulase-negative Staphylococcus (CoNS) are present in the respiratory cavity, particularly in nostrils. We hypothesize that non-epidermidis CoNS found in animals can have the same function as observed in S. epidermidis. Thirty Non-epidermidis CoNS isolates were obtained from poultry, sheep, goat, pig, and dairy cow nostrils. Haemagglutination inhibition (HI) activity was assayed in bacteria-free supernatants from non-epidermidis CoNS against Newcastle disease virus (NDV) and bovine parainfluenza virus type 3 (BPIV). In 13 of the 30 strains (43.3 %), bacteria-free supernatants showed HI activity for NDV and BPIV-3. Staphylococcus xylosus supernatants from poultry (one isolate), sheep (two isolates), goat (one isolate), and dairy cow (three isolates) had the highest frequency of HI activity on NDV and BPIV-3, followed by Staphylococcus sp. supernatants from goat (one isolate), dairy cow (two isolates), and finally Staphylococcus equorum, Staphylococcus chromogens and Staphylococcus gallinarum supernatants with single isolation from poultry, pig and poultry, respectively. Nine isolates had the homologous gene to the embp gene of S. epidermidis, and it was associated with HI activity in the studied viruses. By Pulsed-field gel electrophoresis, S. xylosus isolates showed to be different clones and related to the origin of isolation and HI activity. These results demonstrate that non-epidermidis CoNS supernatants from different animals and origins have the ability of HI on NDV and BPIV-3, indicating that not only S. epidermidis has the same function.

Anaerobic bacteria of the genus Bacteroides are a large group of commensal microorganisms that colonize the human and animal digestive tract. The genus Bacteroides and the closely related genus Parabacteroides include the Bacteroides fragilis group (BFG) of potentially pathogenic bacteria which are frequently isolated from patients with anaerobic infections.The aim of this study was to assess the antimicrobial resistance of environmental strains of the Bacteroides fragilis group. Strains were isolated from human feces, hospital wastewater, influent (UWW) and effluent (TWW) wastewater from a wastewater treatment plant (WWTP), and from the feces of lab rats as a negative control to monitor the entire route of transmission of BFG strains from humans to the environment. The resistance of 123 environmental BFG strains to six antibiotic groups was analyzed with the use of culture-dependent methods. Additionally, the presence of 25 genes encoding antibiotic resistance was determined by PCR.The analyzed environmental BFG strains were highly resistant to the tested antibiotics. The percentage of resistant strains differed between the analyzed antibiotics and was determined at 97.56% for ciprofloxacin, 49.59% for erythromycin, 44.71% for ampicillin, 35.77% for tetracycline, 32.52% for amoxicillin/clavulanic acid, 26.83% for chloramphenicol, 26.01% for clindamycin, 11.38% for moxifloxacin, and 8.94% for metronidazole. The highest drug-resistance levels were observed in the strains isolated from UWW and TWW samples. The mechanisms of antibiotic-resistance were determined in phenotypically resistant strains of BFG. Research has demonstrated the widespread presence of genes encoding resistance to chloramphenicol (100% of all chloramphenicol-resistant strains), tetracyclines (97.78% of all tetracycline-resistant strains), macrolides, lincosamides and streptogramins (81.97% of all erythromycin-resistant strains). Genes encoding resistance to β-lactams and fluoroquinolones were less prevalent. None of the metronidazole-resistant strains harbored the gene encoding resistance to nitroimidazoles. BFG strains isolated from UWW and TWW samples were characterized by the highest diversity of antibiotic-resistance genes and were most often drug-resistant and multidrug-resistant.The present study examines the potential negative consequences of drug-resistant and multidrug-resistant BFG strains that are evacuated with treated wastewater into the environment. The transmission of these bacteria to surface water bodies can pose potential health threats for humans and animals; therefore, the quality of treated wastewater should be strictly monitored. Keywords: Bacteroides fragilis group, Antimicrobial susceptibility, Antibiotic-resistance genes, Environmental strains

Tuberculosis continues to be a public health problem in the world, and drug resistance has been a major obstacle in its treatment. Quinoxaline 1,4-di-N-oxide has been proposed as a scaffold to design new drugs to combat this disease. To examine the efficacy of this compound, this study evaluates methyl, ethyl, isopropyl, and n-propyl esters of quinoxaline 1,4-di-N-oxide derivatives in vitro against Mycobacterium tuberculosis (pansusceptible and monoresistant strains). Additionally, the inhibitory effect of esters of quinoxaline 1,4-di-N-oxide on M. tuberculosis gyrase supercoiling was examined, and a stability analysis by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS) was also carried out. Results showed that eight compounds (T-007, T-018, T-011, T-069, T-070, T-072, T-085 and T-088) had an activity similar to that of the reference drug isoniazid (minimum inhibitory concentration (MIC) = 0.12 µg/mL) with an effect on nonreplicative cells and drug monoresistant strains. Structural activity relationship analysis showed that the steric effect of an ester group at 7-position is key to enhancing its biological effects. Additionally, T-069 showed a high stability after 24 h in human plasma at 37 °C.

Tuberculosis continues to be a public health problem in the world, and drug resistance has been a major obstacle in its treatment. Quinoxaline 1,4-di-N-oxide has been proposed as a scaffold to design new drugs to combat this disease. To examine the efficacy of this compound, this study evaluates methyl, ethyl, isopropyl, and n-propyl esters of quinoxaline 1,4-di-N-oxide derivatives in vitro against Mycobacterium tuberculosis (pansusceptible and monoresistant strains). Additionally, the inhibitory effect of esters of quinoxaline 1,4-di-N-oxide on M. tuberculosis gyrase supercoiling was examined, and a stability analysis by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS) was also carried out. Results showed that eight compounds (T-007, T-018, T-011, T-069, T-070, T-072, T-085 and T-088) had an activity similar to that of the reference drug isoniazid (minimum inhibitory concentration (MIC) = 0.12 &micro, g/mL) with an effect on nonreplicative cells and drug monoresistant strains. Structural activity relationship analysis showed that the steric effect of an ester group at 7-position is key to enhancing its biological effects. Additionally, T-069 showed a high stability after 24 h in human plasma at 37 &deg, C.

In the present study, Aeromonas isolates from diseased and healthy farmed rainbow trout (Oncorhynchus mykiss) in Mexico, were characterized phenotypically and identified to species level by using 16S rDNA RFLP-PCR. A total of 50 isolates were included in the study and 10 Aeromonas species identified. The species A. veronii biovar sobria (22%), A. hydrophila (20%) and A. bestiarum (20%) were the most predominant. All isolates (100%) were resistant to cephalothin.

Hospital-acquired infections (HAIs) are infections that develop in the hospital environment and can be acquired by a patient or hospital staff. They are complications that combine diverse risk factors that make an individual susceptible and are frequently caused by endogenous and exogenous bacterial agents. The most commonly studied etiological agents are bacteria and fungi, with the former representing the most common etiological agents reported to the Hospital Epidemiological Surveillance Network (RHOVE) between 2007 and 2012. Among these agents were Acinetobacter baumannii, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, coagulase-negative Staphylococci (CNS), Enterococcus spp., and Streptococcus pneumoniae. Although obligate anaerobic bacteria are also etiological agents of HAIs, clinical laboratories do not usually perform bacteriological tests to isolate and identify these bacteria. As a result, patients are at a greater risk of not surviving an infection and the epidemiology of this bacterial group is unknown. An important problem associated with HAIs is bacterial multiple drug resistance, which not only increases morbidity and mortality but also the cost of inpatient care. The aim of this review is to provide current information to healthcare professionals on the status of HAIs in Mexico with an emphasis on the etiology, diagnosis, and antimicrobial resistance.

There are major concerns worldwide regarding sexually transmitted infections (STI), bacterial vaginosis (BV), and candidiasis as a major cause of morbidity as they result in significant health and economic consequences, particularly in developing countries. This study was intended to obtain information about the prevalence of these pathologies in women considered to be at low risk using both traditional and in-house NAAT methods. Cervical and vaginal samples were collected all volunteers signed an informed consent form and completed a survey. BV, trichomoniasis, candidiasis, genital mycoplasmas colonization and Chlamydia trachomatis or Neisseria gonorrhoeae cervicitis were diagnosed. Candidiasis and abnormal vaginal flora associated with BV were very frequent. The high colonization with micoplasmas was detected. C. trachomatis cervicitis was found in 10.67% from which a third of the cases were of asymptomatic woman. No cases of gonorrhoea or trichomoniasis were diagnosed. In house NAAT’s used seems to viable tools for the cheap and reliable test for the diagnosis of gonorrhoea and chlamydial infections. Increased awareness of the importance of protected sexual intercourse is imperative to prevent the transmission of sexually transmitted. Further studies for a comprehensive understanding of the rates of these infections in Mexican women are necessary and should be an impulse for make community-based assessment of STI and RTI.

Selective screening media for the detection and identification of Aeromonas strains are needed to guide primary isolation procedures in the clinical laboratory. This study compared the selective CromoCen® AGN chromogenic agar medium for the detection and identification of Aeromonas strains that were isolated from various samples against the conventional selective agar media that are commonly used for the isolation of this organism in food, environmental and clinical samples. The Miles and Misra and ecometric methods were used to evaluate the microbiological performance of CromoCen® AGN chromogenic agar medium, which was shown to be satisfactory. A total of 14 reference Aeromonas strains, 44 wild strains and 106 clinical stool specimens were examined using both non-chromogenic selective agars that are commonly used for Aeromonas isolation and CromoCen® AGN agar. The latter exhibited 94.73% sensitivity and 100% specificity for the various samples. On CromoCen® AGN agar medium, Aeromonas formed colonies with light green, greenish and salmon pigments with or without a surrounding wide transparent zone (halo) of 2–3 mm in diameter around the entire border. This medium is recommended for the isolation and potential identification of the Aeromonas genus.

In contrast to other countries, Mexico lacks precise estimations on the prevalence and risk factors associated with the Chlamydia trachomatis bacterium as well as the impact caused by this infection in different groups of women. There are multiple difficulties, but one of the main reasons is, without a doubt, that cervicovaginal cervicitis caused by this bacterium is not mandatorily reported to epidemiology departments. On the other hand, lymphogranuloma venereal (LGV) is mandatorily reported (including the number of female/male cases, federal entities/districts and age) and the diagnosis is performed using the syndromic management recommended by the World Health Organization (WHO), which is a medical practice with low sensitivity and specificity. A second cause could be the relative difficulty in establishing a diagnosis, considering the laboratory methodology is technically demanding and expensive. A literature review was performed of Mexican research studies on this bacterium, which were presented in congresses or publications, allowing for the identification of a very important number of studies on female patients who visit different governmental and private facilities. These studies report the laboratory methods used to perform diagnostics; furthermore some of these evaluate the sensitivity and specificity. The frequency of this bacterium is varying, which most likely results from the diverse methodologies applied in the cited works. The latest estimations/projections published by the WHO in 2008 indicate that the number of treatable sexually transmitted infection (STI) cases in adults is approximately 498 million cases, and only 105.7 million cases correspond to Chlamydia. Americas accounted for 25.2 million of these cases. Nevertheless, the WHO acknowledges that “There is, however, a great deal of uncertainty surrounding the global and regional STI estimates. If these uncertainties are to be reduced a determined effort is needed to obtain relevant data, in particular: prevalence data disaggregated by age and sex and estimates of the duration of infection disaggregated by pathogen, the health care seeking behaviour of the population, and access to health care”. While C. trachomatis is the most common bacterial cause of Sexually transmitted infections (STIs) in the United States, Mexico provides little to the estimations reported by the WHO.

A comparative study of 109 Aeromonas clinical isolates belonging to the 3 species most frequently isolated from patients with diarrhea in Mexico and Spain was performed to investigate the distribution of 3 prominent toxin genes and the gene encoding flagellin of lateral flagella; 4 well-established virulence factors in the genus Aeromonas. The aerolysin–hemolysin toxin genes were the most prevalent, being present in 89% of the total isolates. The ast toxin gene was conspicuously absent from the Aeromonas caviae and Aeromonas veronii groups but was present in 91% of the Aeromonas hydrophila isolates. Both the alt toxin gene and the lafA flagellin gene also had a low incidence in A. caviae and A. veronii. Differences in the prevalence of alt and lafA were observed between isolates from Mexico and Spain, confirming genus heterogeneity according to geographic location. Carriage of multiple toxin genes was primarily restricted to A. hydrophila isolates, suggesting that A. caviae and A. veronii isolates circulating in Mexico and Spain possess a limited array of virulence genes. Enterobacterial repetitive intergenetic consensus – polymerase chain reaction showed that the Aeromonas populations sampled lack dominant clones and were genetically heterogeneous, with A. caviae being the most diverse species. Further surveys of virulence determinants in genetically heterogeneous populations of Aeromonas isolates circulating worldwide are required to enhance the understanding of their capacity to cause disease.

Aeromonas hydrophila strains recovered from clinical samples and ambient sources were phenotypically and genetically identified. In addition, the distribution of putative virulence factors was assayed. To determine the genetic diversity of these strains, random amplification of polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus (ERIC)-PCR markers were used. The discriminatory ability of the techniques, using Simpsons index, was 0.96 for both methods. The most consistent dendrogram was obtained when RAPD and ERIC data were combined. The genetic diversity revealed a high intra-specific genetic diversity (h = 0.364 ± 0.024 and I = 0.538 ± 0.030). The strains showed a tendency to cluster according to their origin of isolation (bestcut test 0.80 and bootstrap values >50%). The present study demonstrates and quantifies the high intra-specific diversity within this species and reveals a clear differentiation of strains according to their ecological origin. The distribution of virulence-related genes confirm that A. hydrophila is a genetically heterogeneous species that harbour ecotypes which have different pathogenic potential to human and other animals. 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

The aim of this study was to investigate the presence of markers of pathogenicity islands that may be informative to detect the virulent PAI carriers of clinical and environmental strains of Aeromonas spp. isolated in Mexico. virB2, virB9 and virB11 genes were found in Aeromonas strains isolated from environmental and clinical sources while cagE and tfc16 genes were only in strains of environmental origin. Having performed the wide screening presented in this study, we now have a set of strains to map and confirm the presence of a pathogenicity island in Aeromonas strains isolated in Mexico.

Forty-eight Aeromonas isolates from rainbow trout previously identified by the 16S rDNA-RFLP technique were re-identified using 2 housekeeping genes (gyrB and rpoD). After sequencing the prevalences of the species were A. veronii (29.2%), A. bestiarum (20.8%), A. hydrophila (16.7%), A. sobria (10.4%), A. media (8.3%), A. popoffii (6.2%), A. allosaccharophila (2.1%), A. caviae (2.1%), A. salmonicida (2.1%) and one isolate (2.1%) belongs to a candidate new species "Aeromonas lusitana". Coincident identification results to the 16S rDNA-RFLP technique were only obtained for 68.8% of the isolates. PCR amplification of the enterobacterial repetitive intergenic consensus (ERIC-PCR) indicated that the 48 isolates belonged to 33 different ERIC genotypes. Several genotypes were isolated from different farms and organs in the same fish, indicating a systemic dissemination of the bacteria. The presence of genes (blaIMP, blaCphA/IMIS, blaTEM, blaSHV and intI1) that encode extended-spectrum beta-lactamases (ESBLs), metallo-beta-lactamases (MBLs) and class 1 integrons were studied by PCR. Only 39.6% (19/48) of the strains showed the presence of one or more resistance genes. The gene blaCphA/IMIS was detected in 29.2% of the isolates, followed by the intI1 (6.2%) and blaSHV (4.2%) genes. The variable region of class 1 integrons of the 3 positive isolates was sequenced revealing the presence of the gene cassette aadA1 (aminoglycoside transferase) that plays a role in streptomycin/spectinomycin resistance.

Se aisló una cepa de Aeromonas hydrophila perteneciente al grupo de hibridación 3 (HG3) como único patógeno en una muestra de heces diarreicas proveniente de un niño de 1 año de edad que habita en el poblado de Acaxochitlán, Hidalgo. A nivel mundial, el aislamiento de cepas del HG3 (fenotipo A. hydrophila) a partir de heces diarreicas se reporta con una muy baja incidencia y el aislamiento que reportamos en el presente artículo es el primer reporte en México. La cepa se identificó por pruebas bioquímicas y por métodos genéticos (RFLP-16S rDNA y por secuenciación del 16S rDNA). Adicionalmente, se determinó su capacidad de producir diversas enzimas extracelulares relacionadas con la virulencia y se detectaron, vía PCR, los genes que las producen. En el aislamiento se detectaron fenotípica y genéticamente la aerolisina/ hemolisina, la enterotoxina citotónica lábil (alt), las lipasas y las DNasas. Mientras que los genes de la enterotoxina citotónica termoestable (ast) y el flagelo lateral (laf) no se detectaron. La cepa mostró susceptibilidad a cefalosporinas y quinolonas. En el presente artículo se documenta el potencial virulento de la cepa aislada y la posible importancia clínica de cepas de esta genoespecie. A pesar de la aparente baja frecuencia de aislamiento de cepas A. hydrophila del HG 3, éstas no deben subestimarse, ya que en general muestran el mismo potencial virulento y perfil de resistencia que las especies de Aeromonas que exhiben mayores frecuencias de aislamiento. El presente trabajo aporta información al conocimiento de estas cepas tan particulares.

Background Although sophisticated methodologies are available, the use of endpoint polymerase chain reaction (PCR) to detect 16S rDNA genes remains a good approach for estimating the incidence and prevalence of specific infections and for monitoring infections. Considering the importance of the early diagnosis of sexually transmitted infections (STIs), the development of a sensitive and affordable method for identifying pathogens in clinical samples is needed. Highly specific and efficient primers for a multiplex polymerase chain reaction (m-PCR) system were designed in silico to detect the 16S rDNA genes of four bacteria that cause genital infections, and the PCR method was developed. Methods The Genosensor Probe Designer (GPD) (version 1.0a) software was initially used to design highly specific and efficient primers for in-house m-PCR. Single-locus PCR reactions were performed and standardised, and then primers for each locus in turn were added individually in subsequent amplifications until m-PCR was achieved. Amplicons of the expected size were obtained from each of the four bacterial gene fragments. Finally, the analytical specificity and limits of detection were tested. Results Because they did not amplify any product from non-STI tested species, the primers were specific. The detection limits for the Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis and Ureaplasma urealyticum primer sets were 5.12 × 105, 3.9 × 103, 61.19 × 106 and 6.37 × 105 copies of a DNA template, respectively. Conclusions The methodology designed and standardised here could be applied satisfactorily for the simultaneous or individual detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis and Ureaplasma urealyticum. This method is at least as efficient as other previously described methods; however, this method is more affordable for low-income countries.

"The isolation of Aeromonas bestiarum from common carps (Cyprinus carpio L.), cultivated at Santa Maria Chapa de Mota, is reported for the first time. The genetic identification for differentiating A. bestiarum from A. salmonicida is here emphasized."

Administration of empirical antibiotic therapy prior to microbiological diagnosis is thought to be associated the failure of subsequent bacterial growth in culture. The aim of this study was to detect bacterial pathogens via direct amplification and sequencing of the 16S rDNA gene in samples showing negative culture results as alternative diagnostic tools to troubleshoot difficult samples. Twenty-three (7.66 %) positive samples were detected, most of which were monomicrobial infections; 15 of the cases were identified as HAIs, 6 had catheter colonisation, and 2 had sample colonisation. The pathogens identified included Escherichia, Salmonella, Pseudomonas spp., Enterococcus spp. and coagulase-negative staphylococci (CoNS). The most frequent infections were bacteraemia and urinary tract infection, but meningitis, warm infection and soft tissue infection were also documented. These findings emphasise the efficacy and usefulness of molecular diagnosis, thus 16S rDNA gene analysis is strongly indicated by HAIs diagnostics. Electronic supplementary material The online version of this article (doi:10.1186/s40064-016-1949-3) contains supplementary material, which is available to authorized users.

The isolation of Aeromonas bestiarum from common carps (Cyprinus carpio L.), cultivated at Santa Maria Chapa de Mota, is reported for the first time. The genetic identification for differentiating A. bestiarum from A. salmonicida is here emphasized.