ABSTRACT Neonatal sepsis leads to severe morbidity and occasionally death among neonates within the first week following birth, particularly in low- and middle-income countries. Empirical therapy includes antibiotics recommended by WHO. However, these have been ineffective against antimicrobial multidrug-resistant bacterial strains such as Klebsiella spp, Escherichia coli, and Staphylococcus aureus species. To counter this problem, new molecules and alternative sources of compounds with antibacterial activity are sought as options. Actinobacteria, particularly pathogenic strains, have revealed a biotechnological potential still underexplored. This study aimed to determine the presence of biosynthetic gene clusters and the antimicrobial activity of actinobacterial strains isolated from clinical cases against multidrug-resistant bacteria implicated in neonatal sepsis. In total, 15 strains isolated from clinical cases of actinomycetoma were used. PCR screening for the PKS-I, PKS-II, NRPS-I, and NRPS-II biosynthetic systems determined their secondary metabolite-producing potential. The strains were subsequently assayed for antimicrobial activity by the perpendicular cross streak method against Escherichia fergusonii Sec 23, Klebsiella pneumoniae subsp. pneumoniae H1064, Klebsiella variicola H776, Klebsiella oxytoca H793, and Klebsiella pneumoniae subsp. ozaenae H7595, previously classified as multidrug-resistant. Finally, the strains were identified by 16S rRNA gene sequence analysis. It was found that 100% of the actinobacteria had biosynthetic systems. The most frequent biosynthetic system was NRPS-I (100%), and the most frequent combination was NRPS-I and PKS-II (27%). All 15 strains showed antimicrobial activity. The strain with the highest antimicrobial activity was Streptomyces albus 94.1572, as it inhibited the growth of the five multidrug-resistant bacteria evaluated. birth low middleincome middle income countries WHO However multidrugresistant multidrug resistant spp coli species problem options Actinobacteria underexplored total 1 used PKSI, PKSI PKS I, I PKS-I PKSII, PKSII II, II NRPSI, NRPSI NRPS NRPSII metaboliteproducing metabolite producing 23 subsp H1064 H H776 H793 H7595 multidrugresistant. resistant. Finally S analysis 100 100%, , (100%) 27%. 27 27% . (27%) 941572 94 1572 94.1572 evaluated 2 H106 H77 H79 H759 10 (100% (27% 94157 9 157 94.157 H10 H7 H75 (100 (27 9415 94.15 H1 (10 (2 941 94.1 (1 ( 94.

ABSTRACT Mycetoma is a neglected tropical disease (NTD) declared by the World Health Organization (WHO) in 2016. It is characterized by the progressive growth of nodules and granulomatous lesions on the legs, arms, and trunk. It is potentially disfiguring and causes disability or amputations in working-age people from marginalized areas. The causative agents can be fungi (eumycetoma) or actinobacteria (actinomycetoma), the latter being the most common in America and Asia. Nocardia brasiliensis is the most important causal agent of actinomycetoma in the Americas. Taxonomic problems have been reported when identifying this species, so this study aimed to detect the 16S rRNA gene variations in N. brasiliensis strains using an in silico enzymatic restriction technique. The study included strains from clinical cases of actinomycetoma in Mexico, isolated from humans and previously identified as N. brasiliensis by traditional methods. The strains were characterized microscopically and macroscopically, then subjected to DNA extraction and amplification of the 16S rRNA gene by PCR. The amplification products were sequenced, and consensus sequences were constructed and used for genetic identification and in silico restriction enzyme analysis with the New England BioLabs® NEBcutter program. All study strains were molecularly identified as N. brasiliensis; however, in silico restriction analysis detected a diversity in the restriction patterns that were finally grouped and subclassified into 7 ribotypes. This finding confirms the existence of subgroups within N. brasiliensis. The results support the need to consider N. brasiliensis as a complex species. NTD (NTD WHO (WHO 2016 legs arms trunk workingage working age areas eumycetoma (eumycetoma actinomycetoma, , (actinomycetoma) Asia Americas species S N technique Mexico methods macroscopically PCR sequenced BioLabs program however ribotypes 201 (actinomycetoma 20 2

ABSTRACT Heavy metals released into the environment cause deterioration of environmental health. Their removal by biological means is a strategy under study. This research aimed to isolate and identify halophilic bacteria from saline-sodic soils and determine their tolerance to the metalloid As3+ and the heavy metals Cr6+, Hg2+, and Pb2+. The strains were grown on halophilic medium (HM), without and with the presence of each metal, at 10% NaCl, pH 8.0 ± 0.2, and 37 ºC. The strains with the highest tolerance to each metal were identified by 16S rRNA gene sequencing and phylogenetic analysis. The minimum inhibitory concentration (MIC) was subsequently determined. Seventy-five halophilic strains were isolated. The isolates with the highest tolerance were Salinicococcus spp. strains TX3SA-2MHG1 and TX3SA-4MHG1 [MIC (Hg2+) of 0.1 mmol/L]; Halomonas sp. strain TXO4B-1SG9 [MIC (Pb2+) of 7.0 mmol/L]; Nocardiopsis sp. strains TXO7B-1SG12 and TXV10-3SG5 [MIC (As3+) of 8.5 mmol/L], and Nocardiopsis sp. strain TXV7-8SG2 [MIC (As3+) of 27.25 mmol/L; MIC (Cr6+) 1250.0 mmol/L; MIC (Hg2+) 0.075 mmol/L and MIC (Pb2+) 7.5 mmol/L]. It was possible to obtain and identify isolates of halophilic metal tolerant bacteria.

RESUMEN Los metales pesados liberados en el ambiente causan el deterioro de la salud ambiental. Su eliminación por medios biológicos es una estrategia en estudio. El objetivo de esta investigación fue aislar e identificar bacterias halófilas a partir de suelos salino-sódicos y determinar su tolerancia al metaloide As3+ y a los metales pesados Cr6+, Hg2+ y Pb2+. Las cepas fueron cultivadas en medio halófilo (MH), sin y con la presencia de cada metal, al 10 % de NaCl, pH 8.0 ± 0.2 y 37 ºC. Las cepas que mostraron mayor tolerancia a cada metal se identificaron por análisis de secuenciación del gen del ARNr 16S y por análisis filogenético, posteriormente se determinó la concentración mínima inhibitoria (CMI). Se aislaron 75 cepas halófilas. Los aislados con mayor tolerancia fueron Salinicoccus spp. TX3SA-2MHG1 y TX3SA-4MHG1 [CMI (Hg2+) de 0.1 mmol/L]; Halomonas sp. TXO4B-1SG9 [CMI (Pb2+) de 7.0 mmol/L]; Nocardiopsis sp. TXO7B-1SG12 y TXV10-3SG5 [CMI (As3+) de 8. 5 mmol/L] y Nocardiopsis sp. TXV7-8SG2 [CMI (As3+) de 27.25 mmol/L; CMI (Cr6+) 1 250.0 mmol/L; CMI (Hg2+) 0.075 mmol/L y CMI (Pb2+) 7.5 mmol/L]. Fue posible obtener e identificar aislados de bacterias halófilas metalo-tolerantes.

OBJECTIVE: The purpose of this paper is to describe some of the results of the Morelos HPV Study. The main objective of the Morelos HPV Study is to evaluate the use of human papillomavirus (HPV) DNA testing, as compared to the Papanicolaou (Pap) test, for cervical cancer (CC) screening. MATERIAL AND METHODS: The Morelos HPV Study is currently being conducted in Mexico, to examine the possibility of using HPV testing for CC screening. The HPV testing of self-collected vaginal and clinician-collected cervical specimens was evaluated as part of this study. The acceptability of the HPV testing of self-collected specimens was compared to that of the Pap test. A cost-effectiveness analysis (CEA) and cost-benefit analysis (CBA) was also performed. RESULTS: The Morelos HPV Study results indicate that HPV testing has a greater sensitivity to detect cervical intraepithelial neoplasia (CIN) 2/3 and CC than the Pap test. Our results also indicate an over-all lower acceptability of the Pap test as compared to the self-collected procedure. The results of the CEA and CBA indicate that screening women between the ages of 20-80 for CC using some type of HPV testing is always more cost-effective than screening for CC using the Pap test. CONCLUSIONS: Our results suggest that self- and clinician-collected HPV testing could be used in CC prevention programs, as an effective complement or substitute for the Pap test.

OBJETIVO: Describir algunos de los resultados del Estudio de VPH en Morelos. El objetivo principal del Estudio de VPH en Morelos es evaluar el uso de la prueba del virus de papiloma humano (VPH), en relación con la prueba de Papanicolaou, para el tamizaje de cáncer cervical. MATERIAL Y MÉTODOS: El Estudio de VPH en Morelos actualmente se está llevando a cabo en México, para examinar la posibilidad de usar la prueba de VPH para la detección de cáncer cervical. Se evaluó el uso de la prueba de VPH en muestras auto-tomadas vaginales y en muestras cervicales tomadas por un clínico. Se comparó la aceptabilidad del uso de la prueba de VPH en muestras auto-tomadas al uso del Papanicolaou. También se realizó un análisis de costo-efectividad y de costo-beneficio. RESULTADOS: Los resultados del Estudio de VPH en Morelos indican que la prueba de VPH tiene una mayor sensibilidad para detectar los casos de neoplasia intraepitelial cervical 2/3 y cáncer cervical que la prueba de Papanicolaou. Los resultados también indican una aceptabilidad menor al uso de la prueba de Papanicolaou que al uso de la prueba de VPH auto-tomada. Los resultados del análisis de costo-efectividad y el análisis de costo-beneficio indican que el tamizaje con la prueba de VPH en mujeres de 20-80 años de edad siempre es más costo-efectivo que el tamizaje con el Papanicolaou. CONCLUSIONES: Nuestros resultados sugieren que la prueba del VPH (ya sea auto-tomada o clínica) podría ser utilizada en los programas de detección y prevención de cáncer cervical, como un complemento o un sustituto efectivo de la prueba de Papanicolaou.

OBJECTIVE: The purpose of this paper is to describe some of the results of the Morelos HPV Study. The main objective of the Morelos HPV Study is to evaluate the use of human papillomavirus (HPV) DNA testing, as compared to the Papanicolaou (Pap) test, for cervical cancer (CC) screening. MATERIAL AND METHODS: The Morelos HPV Study is currently being conducted in Mexico, to examine the possibility of using HPV testing for CC screening. The HPV testing of self-collected vaginal and clinician-collected cervical specimens was evaluated as part of this study. The acceptability of the HPV testing of self-collected specimens was compared to that of the Pap test. A cost-effectiveness analysis (CEA) and cost-benefit analysis (CBA) was also performed. RESULTS: The Morelos HPV Study results indicate that HPV testing has a greater sensitivity to detect cervical intraepithelial neoplasia (CIN) 2/3 and CC than the Pap test. Our results also indicate an over-all lower acceptability of the Pap test as compared to the self-collected procedure. The results of the CEA and CBA indicate that screening women between the ages of 20-80 for CC using some type of HPV testing is always more cost-effective than screening for CC using the Pap test. CONCLUSIONS: Our results suggest that self- and clinician-collected HPV testing could be used in CC prevention programs, as an effective complement or substitute for the Pap test.

OBJETIVO: Describir algunos de los resultados del Estudio de VPH en Morelos. El objetivo principal del Estudio de VPH en Morelos es evaluar el uso de la prueba del virus de papiloma humano (VPH), en relación con la prueba de Papanicolaou, para el tamizaje de cáncer cervical. MATERIAL Y MÉTODOS: El Estudio de VPH en Morelos actualmente se está llevando a cabo en México, para examinar la posibilidad de usar la prueba de VPH para la detección de cáncer cervical. Se evaluó el uso de la prueba de VPH en muestras auto-tomadas vaginales y en muestras cervicales tomadas por un clínico. Se comparó la aceptabilidad del uso de la prueba de VPH en muestras auto-tomadas al uso del Papanicolaou. También se realizó un análisis de costo-efectividad y de costo-beneficio. RESULTADOS: Los resultados del Estudio de VPH en Morelos indican que la prueba de VPH tiene una mayor sensibilidad para detectar los casos de neoplasia intraepitelial cervical 2/3 y cáncer cervical que la prueba de Papanicolaou. Los resultados también indican una aceptabilidad menor al uso de la prueba de Papanicolaou que al uso de la prueba de VPH auto-tomada. Los resultados del análisis de costo-efectividad y el análisis de costo-beneficio indican que el tamizaje con la prueba de VPH en mujeres de 20-80 años de edad siempre es más costo-efectivo que el tamizaje con el Papanicolaou. CONCLUSIONES: Nuestros resultados sugieren que la prueba del VPH (ya sea auto-tomada o clínica) podría ser utilizada en los programas de detección y prevención de cáncer cervical, como un complemento o un sustituto efectivo de la prueba de Papanicolaou.