A bioinformatic approach to identify confirmed and probable CRISPR-Cas systems in the Acinetobacter calcoaceticus-Acinetobacter baumannii complex genomes.

Introduction: The Acinetobacter calcoaceticus-Acinetobacter baumannii complex, or Acb complex, consists of six species: Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter nosocomialis, Acinetobacter pittii, Acinetobacter seifertii, and Acinetobacter lactucae. A. baumannii is the most clinically significant of these species and is frequently related to healthcare-associated infections (HCAIs). Clustered regularly interspaced short palindromic repeat (CRISPR) arrays and associated genes (cas) constitute bacterial adaptive immune systems and function as variable genetic elements. This study aimed to conduct a genomic analysis of Acb complex genomes available in databases to describe and characterize CRISPR systems and cas genes. Methods: Acb complex genomes available in the NCBI and BV-BRC databases, the identification and characterization of CRISPR-Cas systems were performed using CRISPRCasFinder, CRISPRminer, and CRISPRDetect. Sequence types (STs) were determined using the Oxford scheme and ribosomal multilocus sequence typing (rMLST). Prophages were identified using PHASTER and Prophage Hunter. Results: A total of 293 genomes representing six Acb species exhibited CRISPR-related sequences. These genomes originate from various sources, including clinical specimens, animals, medical devices, and environmental samples. Sequence typing identified 145 ribosomal multilocus sequence types (rSTs). CRISPR-Cas systems were confirmed in 26.3% of the genomes, classified as subtypes I-Fa, I-Fb and I-Fv. Probable CRISPR arrays and cas genes associated with CRISPR-Cas subtypes III-A, I-B, and III-B were also detected. Some of the CRISPR-Cas systems are associated with genomic regions related to Cap4 proteins, and toxin-antitoxin systems. Moreover, prophage sequences were prevalent in 68.9% of the genomes. Analysis revealed a connection between these prophages and CRISPR-Cas systems, indicating an ongoing arms race between the bacteria and their bacteriophages. Furthermore, proteins associated with anti-CRISPR systems, such as AcrF11 and AcrF7, were identified in the A. baumannii and A. pittii genomes. Discussion: This study elucidates CRISPR-Cas systems and defense mechanisms within the Acb complex, highlighting their diverse distribution and interactions with prophages and other genetic elements. This study also provides valuable insights into the evolution and adaptation of these microorganisms in various environments and clinical settings.

Characterization of Cronobacter sakazakii and Cronobacter malonaticus Strains Isolated from Powdered Dairy Products Intended for Consumption by Adults and Older Adults.

The objective of this study was to characterize Cronobacter spp. and related organisms isolated from powder dairy products intended for consumption by adults and older adults using whole-genome sequencing (WGS), and to identify genes and traits that encode antibiotic resistance and virulence. Virulence (VGs) and antibiotic resistance genes (ARGs) were detected with the Comprehensive Antibiotic Resistance Database (CARD) platform, ResFinder, and MOB-suite tools. Susceptibility testing was performed using disk diffusion. Five presumptive strains of Cronobacter spp. were identified by MALDI-TOF MS and ribosomal MLST. Three C. sakazakii strains were of the clinical pathovar ST1, one was ST31, and the remaining isolate was C. malonaticus ST60. In addition, Franconibacter helveticus ST345 was identified. The C. sakazakii ST1 strains were further distinguished using core genome MLST based on 2831 loci. Moreover, 100% of the strains were resistant to cefalotin, 75% to ampicillin, and 50% to amikacin. The C. sakazakii ST1 strains were multiresistant (MDR) to four antibiotics. Additionally, all the strains adhered to the N1E-115 cell line, and two invaded it. Eighteen ARGs mainly involved in antibiotic target alteration and antibiotic efflux were detected. Thirty VGs were detected and clustered as flagellar proteins, outer membrane proteins, chemotaxis, hemolysins, and genes involved in metabolism and stress. The pESA3, pSP291-1, and pCMA1 plasmids were detected, and the prevalent mobile genetic elements (MGEs) were ISEsa1, ISEc52, and IS26. The isolates of C. sakazakii and C. malonaticus exhibited multiresistance to antibiotics, harbored genes encoding various antibiotic resistance proteins, and various virulence factors. Consequently, these contaminated powdered dairy products pose a risk to the health of hypersensitive adults.

Screening of Antibiotic and Virulence Genes from Whole Genome Sequenced Cronobacter sakazakii Isolated from Food and Milk-Producing Environments.

The objective of this study was to use whole-genome sequencing (WGS) to screen for genes encoding for antibiotic resistance, fitness and virulence in Cronobacter sakazakii strains that had been isolated from food and powdered-milk-producing environments. Virulence (VGs) and antibiotic-resistance genes (ARGs) were detected with the Comprehensive Antibiotic Resistance Database (CARD) platform, ResFinder and PlasmidFinder tools. Susceptibility testing was performed using disk diffusion. Fifteen presumptive strains of Cronobacter spp. were identified by MALDI-TOF MS and ribosomal-MLST. Nine C. sakazakii strains were found in the meningitic pathovar ST4: two were ST83 and one was ST1. The C. sakazakii ST4 strains were further distinguished using core genome MLST based on 3678 loci. Almost all (93%) strains were resistant to cephalotin and 33% were resistant to ampicillin. In addition, 20 ARGs, mainly involved in regulatory and efflux antibiotics, were detected. Ninety-nine VGs were detected that encoded for OmpA, siderophores and genes involved in metabolism and stress. The IncFIB (pCTU3) plasmid was detected, and the prevalent mobile genetic elements (MGEs) were ISEsa1, ISEc52 and ISEhe3. The C. sakazakii isolates analyzed in this study harbored ARGs and VGs, which could have contributed to their persistence in powdered-milk-producing environments, and increase the risk of infection in susceptible population groups.

Are Enterobacteriaceae and Enterococcus Isolated from Powdered Infant Formula a Hazard for Infants? A Genomic Analysis.

Powdered infant formulas (PIF) are the most used dietary substitutes that are used in order to supplement breastfeeding. However, PIF are not sterile and can be contaminated with different microorganisms. The objective of this study was to genomically characterize Enterobacteriaceae (ENT) and Enterococcus strains that were isolated from PIF. Strains were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and whole-genome sequencing (WGS). Genomic typing, detection of virulence, and resistance profiles and genes were performed with the Ridom SeqSphere+ software; the comprehensive antibiotic resistance database (CARD) platform; ResFinder and PlasmidFinder tools; and by the disk diffusion method. Nineteen isolates from PIF were analyzed, including ENT such as Kosakonia cowanii, Enterobacter hormaechei, Franconibacter helveticus, Mixta calida, and lactic acid bacteria such as Enterococcus faecium. The strains exhibited resistance to beta-lactams, cephalosporins, and macrolides. Resistance genes such as AcrAB-TolC, marA, msbA, knpEF, oqxAB, fosA, blaACT-7, blaACT-14,qacJ, oqxAB,aac(6')-Ii, and msr(C); and virulence genes such as astA, cheB, cheR, ompA ompX, terC, ironA, acm, and efaAfm, adem were also detected. All the analyzed strains possessed genes that produced heat-shock proteins, such as IbpA and ClpL. In PIF, the presence of ENT and Enterococcus that are multiresistant to antibiotics-together with resistance and virulence genes-pose a health risk for infants consuming these food products.

Maternal Factors Associated with Non-Exclusive Breastfeeding in Haitian Immigrant Women in Southern Chile.

There is limited knowledge concerning factors that affect non-exclusive breastfeeding (NEBF) practices in immigrant populations, especially in Latin America. The objective of the present study was to determine the association between maternal factors and the prevalence of NEBF in Haitian immigrant women in southern Chile. This was an analytical cross-sectional study. The probabilistic sample consisted of 173 Haitian women who gave informed consent. Sociodemographic and dietary-nutritional information was collected from all participants. Bivariate (χ2) and multivariate (logistic regression) inferential statistics were applied. All analyses were performed with the STATA 16.0 statistical software, and the significance level was established as α < 0.05. The prevalence of EBF at 6 mo was 54.3%. Maternal factors associated with a lower prevalence of EBF were not having permanent residency (OR: 2.34, CI: 2.18−2.83), residency <12 months (OR: 2.23, CI: 2.09−2.78), limited knowledge of breastfeeding (OR: 1.96; CI: 1.81−2.27), and low educational attainment (OR: 1.78; CI: 1.61−2.11). The protective factors were employment (OR: 0.36, CI: 0.28−0.40), access to basic services (OR: 0.32; CI: 0.22−0.48), and Spanish proficiency (OR: 0.29; CI: 0.20−0.51). Haitian immigrant women without legal residency, recently arrived, with low educational attainment, and poor knowledge of breastfeeding have more risk of not providing exclusive breastfeeding. Targeted interventions for mothers with these risk factors may help improve EBF rates.

Malnutrición por exceso en niños de 5 a 10 años y su asociación con el estado nutricional pre y gestacional, lactancia materna y patología materna

La malnutrición por exceso es un problema de salud pública relevante, con origen multifactorial y factores asociados como el estado nutricional pregestacional (ENP), durante y post gestación, ganancia excesiva de peso (EGP) gestacional y duración de la lactancia materna. El objetivo del estudio fue evaluar la malnutrición por exceso en niños de 5 a 10 años y su asociación con el estado nutricional pre y gestacional, lactancia materna exclusiva (LME) y patologías maternas. Se realizó un estudio con 213 niños y niñas estudiantes de 5 a 10 años. Los datos biosociodemográficos se obtuvieron mediante entrevistas con las madres y datos maternos previos y el embarazo con el carné de control. El estimó estado nutricional se estimó con z-score del peso para la talla e IMC/E, según sexo. Para los análisis estadísticos se usó el software STATA 15.0. El 56% de los estudiantes tuvo malnutrición por exceso. Un 53,1% de las madres presentó malnutrición por exceso previo al embarazo que aumentó a 74,6% post gestación. El 71,4% de los niños tuvo una LME ≥6 meses. Se relacionó estadísticamente la edad materna ≥35 años (p= 0,044) y la malnutrición por exceso pregestacional (p= 0,014). La edad materna ≥35 años aumentó el riesgo de malnutrición por exceso casi dos veces (OR= 1,78; IC: 1,029-3,046), al igual que el ENP (OR= 2,11; IC: 1,193-3,693) y en patologías maternas (OR= 1,41; IC: 1, 073-2,694). En conclusión, los niños preescolares de 5 a 10 años con factores de edad materna ≥35, ENP y patologías maternas tuvieron entre 1,4 y 2,11 veces más riesgo de presentar malnutrición por exceso comparado con aquellos niños sin estos factores.

Overnutrition is a relevant public health problem with a multifactorial origin. Associated factors include maternal nutritional status before, during, and after gestation, excessive gestational weight gain, and breastfeeding duration. The objective of the study was to evaluate the association between overnutrition in children aged 5-10 years and pre- and gestational nutritional status, exclusive breastfeeding, and maternal pathologies. The study consisted of 213 schoolchildren aged 5-10 years. Bio-socio-demographic data were obtained through interviews with mothers, previous maternal data and pregnancy follow-up records. Nutritional status was measured using weight-for-height and BMI z-score by age and sex. The STATA 15.0 software was used for statistical analysis. A total of 56% of the schoolchildren had overnutrition. Meanwhile, 53.1% of the mothers demonstrated pre-pregnancy overnutrition, which increased to 74.6% post-pregnancy. It was found that 71.4% of schoolchildren had experienced exclusive breastfeeding ≥6 months. Maternal age ≥35 years (p= 0.044) and pre-gestational nutritional status (p= 0.014) were statistically related. Maternal age ≥35 years increased overnutrition by almost two-fold (OR= 1.78; IC: 1.029-3.046), as did pre-gestational nutritional status (OR= 2.11; IC: 1.193-3.693) and maternal pathologies (OR= 1.41; IC: 1.073-2.694). In conclusion, schoolchildren aged 5-10 years with mothers ≥35 years of age, who had overnutrition in the pre-gestational period, and a pathology were 1.4 to 2.44 times more at risk of developing overnutrition than children without these factors.

Association between Maternal Factors, Preterm Birth, and Low Birth Weight of Chilean Singletons.

There has been an increase in preterm (PT) births in Western countries in recent years, which is associated with low-birthweight (LBW) children. The aim of this study was to determine the association between maternal factors and PT and LBW Chilean newborns. Methods: This was an analytical cross-sectional study of a national sample of 903,847 newborns and their mothers. The newborn gestational age, birth weight, maternal age, marital status, education, employment situation, and residence were analyzed. A multivariate logistic regression model was applied (α = 0.05) (STATA v.15). The prevalence was 6.8% and 5.0% for PT and LBW, respectively. The probability of the newborns being PT and LBW was 1.18 and 1.22 times if their mothers had <12 years of education and 1.38 and 1.29 times if the mothers were ≥35 years old, respectively. Mothers with <12 years education and ≥35 years were risk factors for PT and LBW newborns. Maternal educational attainment was a protective factor for the Chilean newborns, and a maternal age ≥35 years was a risk factor for PT and LBW.

Genomic Characterization of Cronobacter spp. and Salmonella spp. Strains Isolated From Powdered Infant Formula in Chile.

This study characterized five Cronobacter spp. and six Salmonella spp. strains that had been isolated from 155 samples of powdered infant formula (PIF) sold in Chile and manufactured in Chile and Mexico in 2018-2020. Two strains of Cronobacter sakazakii sequence type (ST) ST1 and ST31 (serotypes O:1 and O:2) and one strain of Cronobacter malonaticus ST60 (O:1) were identified. All Salmonella strains were identified as Salmonella Typhimurium ST19 (serotype O:4) by average nucleotide identity, ribosomal multilocus sequence typing (rMLST), and core genome MLST (cgMLST). The C. sakazakii and C. malonaticus isolates were resistant to cephalothin, whereas the Salmonella isolates were resistant to oxacillin and ampicillin. Nineteen antibiotic resistance genes were detected in the C. sakazakii and C. malonaticus isolates; the most prevalent were mcr-9.1, blaCSA , and blaCMA . In Salmonella, 30 genes encoding for aminoglycoside and cephalosporin resistance were identified, including aac(6')-Iaa, ß-lactamases ampH, ampC1, and marA. In the Cronobacter isolates, 32 virulence-associated genes were detected by WGS and clustered as flagellar proteins, outer membrane proteins, chemotaxis, hemolysins, invasion, plasminogen activator, colonization, transcriptional regulator, survival in macrophages, use of sialic acid, and toxin-antitoxin genes. In the Salmonella strains, 120 virulence associated genes were detected, adherence, magnesium uptake, resistance to antimicrobial peptides, secretion system, stress protein, toxin, resistance to complement killing, and eight pathogenicity islands. The C. sakazakii and C. malonaticus strains harbored I-E and I-F CRISPR-Cas systems and carried Col(pHHAD28) and IncFIB(pCTU1) plasmids, respectively. The Salmonella strains harbored type I-E CRISPR-Cas systems and carried IncFII(S) plasmids. The presence of C. sakazakii and Salmonella in PIF is a health risk for infants aged less than 6 months. For this reason, sanitary practices should be reinforced for its production and retail surveillance.

Obese Women Have a High Carbohydrate Intake without Changes in the Resting Metabolic Rate in the Luteal Phase.

Hormonal changes are caused by the menstrual cycle phases, which influence resting metabolic rate and eating behavior. The aim of the study was to determine resting metabolic rate (RMR) and its association with dietary intake according to the menstrual cycle phase in lean and obese Chilean women. This cross-sectional analytical study included 30 adult women (15 lean and 15 with obesity). Body composition was measured with a tetrapolar bioelectrical impedance meter. Nutritional status was determined by adiposity. A 24-h recall of three nonconsecutive days verifies dietary intake. The RMR was measured by indirect calorimetry. All measurements were performed in both the follicular and luteal phases of the menstrual cycle. Statistical analyses were performed with STATA software at a significance level, which was α = 0.05. The RMR (ß = 121.6 kcal/d), temperature (ß = 0.36 °C), calorie intake (ß = 317.1 kcal/d), and intake of lipids (ß = 13.8 g/d) were associated with the luteal phase in lean women. Only extracellular water (ß = 1.11%) and carbohydrate consumption (ß = 45.2 g/d) were associated in women with obesity. Lean women showed increased RMR, caloric intake, and lipid intake during the luteal phase. For women with obesity, carbohydrate intake increased but not RMR.

Association of Prematurity and Low Birth Weight with Gestational Exposure to PM2.5 and PM10 Particulate Matter in Chileans Newborns.

Fetal growth can be affected by gestational exposure to air pollution. The aim of the study was to determine the association between prematurity and low birth weight (LBW) with gestational exposure to PM2.5 and PM10 particulate matter in Chileans newborns. This cross-sectional analytical study included 595,369 newborns. Data were extracted from the live newborn records of the Chilean Ministry of Health. Sex, gestational age, birth weight, and living variables were analyzed. We used the Air Quality Information System of the Chilean Ministry of the Environment to obtain mean PM2.5 and PM10 emissions. A multivariate logistic regression model was performed with STATA 15.0 software at α < 0.05. Prevalence was 7.4% prematurity and 5.5% LBW. Mean PM2.5 and PM10 concentrations were 25.5 µg/m3 and 55.3 µg/m3, respectively. PM2.5 was associated with an increased the risk of LBW (OR: 1.031; 95%CI: 1.004−1.059) when exposure occurred in the second trimester, while PM10 affected the whole pregnancy. In addition, PM10 exposure in any gestational trimester was associated with an increased the risk of prematurity. The PM10 particulate matter was associated with both prematurity and LBW in all of the trimesters of exposure. The PM2.5 particulate matter was only associated with LBW when exposure occurred in the second gestational trimester.

Profiling the Virulence and Antibiotic Resistance Genes of Cronobacter sakazakii Strains Isolated From Powdered and Dairy Formulas by Whole-Genome Sequencing.

Cronobacter sakazakii is an enteropathogen that causes neonatal meningitis, septicemia, and necrotizing enterocolitis in preterm infants and newborns with a mortality rate of 15 to 80%. Powdered and dairy formulas (P-DF) have been implicated as major transmission vehicles and subsequently the presence of this pathogen in P-DF led to product recalls in Chile in 2017. The objective of this study was to use whole genome sequencing (WGS) and laboratory studies to characterize Cronobacter strains from the contaminated products. Seven strains were identified as C. sakazakii, and the remaining strain was Franconibacter helveticus. All C. sakazakii strains adhered to a neuroblastoma cell line, and 31 virulence genes were predicted by WGS. The antibiograms varied between strains. and included mcr-9.1 and bla CSA genes, conferring resistance to colistin and cephalothin, respectively. The C. sakazakii strains encoded I-E and I-F CRISPR-Cas systems, and carried IncFII(pECLA), Col440I, and Col(pHHAD28) plasmids. In summary, WGS enabled the identification of C. sakazakii strains and revealed multiple antibiotic resistance and virulence genes. These findings support the decision to recall the contaminated powdered and dairy formulas from the Chilean market in 2017.

Draft Genome Sequences of Seven Cronobacter sakazakii Strains Carrying the mcr 9.1 Gene Isolated in Chile.

Cronobacter sakazakii is a pathogen that causes severe diseases such as meningitis and necrotizing enterocolitis in infants under 12 months, associated with the consumption of contaminated rehydrated powdered infant formula (PIF). We present seven C. sakazakii genome sequences isolated from PIF and dairy products in Chile in 2017.

Índice de alimentación saludable e indicador sodio/(potasio+calcio) en estudiantes universitarios chilenos / Healthy eating index and sodium/(potassium+calcium) indicator in Chilean university students

Resumen Introducción: Los hábitos alimentarios poco saludables de los estudiantes universitarios constituyen una preocupación importante para la atención medica-nutricional futura. Objetivo: Determinar el Índice de Alimentación Saludable (IAS) e indicador sodio/(potasio+calcio) en universitarios chilenos. Materiales y métodos: Investigación analítica, de corte transversal con una muestra no probabilística de 420 estudiantes, a quienes se aplicó encuesta de información sociodemográfica y antropométrica. Con el cuestionario de frecuencia de consumo y recordatorio de 24 horas se determinó el IAS e indicador sodio/(potasio+calcio). Se aplicó estadística descriptiva e inferencial utilizando el programa Statistical Package for the Social Sciences v22.0. Resultados: El peso, talla e Índice de Masa Corporal, presentaron diferencias estadísticamente significativas, según sexo (p<0,05). Misma situación fue encontrada en el promedio de ingesta de calorías, lípidos, carbohidratos, fibra, calcio y sodio (p<0,05). La ingesta de frutas, verduras y lácteos bajos en grasa fue mayor en mujeres (p<0,01). La mayoría de los sujetos presentaron alimentación poco saludable, siendo mayor en el caso de los hombres (p<0,05). Sólo el 6% de los hombres y 17% de las mujeres tenían una dieta cardio-protectora. Conclusiones: Los estudiantes presentan una dieta con deficiente ingesta de micronutrientes cardioprotectores, hecho que incrementa el riesgo de desarrollar enfermedades cardiovasculares.

Abstract Introduction: Unhealthy eating habits of university students constitutes an important concern for future medical and nutritional care. Objective: To determine the Healthy Eating Index (HEI) and the sodium/(potassium+calcium) indicator in Chilean university students. Materials and methods: An analytical cross-sectional research was carried out with a non-probabilistic sample of 420 students, who answered a survey covering sociodemographic and anthropometric issues. The HEI and sodium/(potassium+calcium) indicator were determined using the consumption frequency and 24-hour reminder questionnaire. A descriptive and inferential statistical analysis was applied through the SPSS (v22.0) program. Results: The weight, height and the body mass index (BMI) of the participants showed statistically significant differences according to their gender (p<0.05). A similar pattern was found in the average intake of calories, lipids, carbohydrates, fiber, calcium and sodium (p<0.05). Intake of fruits, vegetables and low-fat dairy products was higher in women (p<0.01). The majority of participating students had an unhealthy diet, with the situation being more severe in men (p<0.05). Only 6% of men and 17% of women had a cardio-protective diet. Conclusions: Students follow diets with a deficient intake of cardio-protective micronutrients, which increases the risk of developing cardiovascular diseases.

Virulence and Antibiotic Resistance Genes in Listeria monocytogenes Strains Isolated From Ready-to-Eat Foods in Chile.

Listeria monocytogenes is causing listeriosis, a rare but severe foodborne infection. Listeriosis affects pregnant women, newborns, older adults, and immunocompromised individuals. Ready-to-eat (RTE) foods are the most common sources of transmission of the pathogen This study explored the virulence factors and antibiotic resistance in L. monocytogenes strains isolated from ready-to-eat (RTE) foods through in vitro and in silico testing by whole-genome sequencing (WGS). The overall positivity of L. monocytogenes in RTE food samples was 3.1% and 14 strains were isolated. L. monocytogenes ST8, ST2763, ST1, ST3, ST5, ST7, ST9, ST14, ST193, and ST451 sequence types were identified by average nucleotide identity, ribosomal multilocus sequence typing (rMLST), and core genome MLST. Seven isolates had serotype 1/2a, five 1/2b, one 4b, and one 1/2c. Three strains exhibited in vitro resistance to ampicillin and 100% of the strains carried the fosX, lin, norB, mprF, tetA, and tetC resistance genes. In addition, the arsBC, bcrBC, and clpL genes were detected, which conferred resistance to stress and disinfectants. All strains harbored hlyA, prfA, and inlA genes almost thirty-two the showed the bsh, clpCEP, hly, hpt, iap/cwhA, inlA, inlB, ipeA, lspA, mpl, plcA, pclB, oat, pdgA, and prfA genes. One isolate exhibited a type 11 premature stop codon (PMSC) in the inlA gene and another isolate a new mutation (deletion of A in position 819). The Inc18(rep25), Inc18(rep26), and N1011A plasmids and MGEs were found in nine isolates. Ten isolates showed CAS-Type II-B systems; in addition, Anti-CRISPR AcrIIA1 and AcrIIA3 phage-associated systems were detected in three genomes. These virulence and antibiotic resistance traits in the strains isolated in the RTE foods indicate a potential public health risk for consumers.

Molecular Characterization of Cronobacter sakazakii Strains Isolated from Powdered Milk.

Cronobacter spp. are opportunistic pathogens of the Enterobacteriaceae family. The organism causes infections in all age groups, but the most serious cases occur in outbreaks related to neonates with meningitis and necrotizing enterocolitis. The objective was to determine the in silico and in vitro putative virulence factors of six Cronobacter sakazakii strains isolated from powdered milk (PM) in the Czech Republic. Strains were identified by MALDI-TOF MS and whole-genome sequencing (WGS). Virulence and resistance genes were detected with the Ridom SeqSphere+ software task template and the Comprehensive Antibiotic Resistance Database (CARD) platform. Adherence and invasion ability were performed using the mouse neuroblastoma (N1E-115 ATCCCRL-2263) cell line. The CRISPR-Cas system was searched with CRISPRCasFinder. Core genome MLST identified four different sequence types (ST1, ST145, ST245, and ST297) in six isolates. Strains 13755-1B and 1847 were able to adhere in 2.2 and 3.2 × 106 CFU/mL, while 0.00073% invasion frequency was detected only in strain 1847. Both strains 13755-1B and 1847 were positive for three (50.0%) and four virulence genes, respectively. The cpa gene was not detected. Twenty-eight genes were detected by WGS and grouped as flagellar or outer membrane proteins, chemotaxis, hemolysins, and invasion, plasminogen activator, colonization, transcriptional regulator, and survival in macrophages. The colistin-resistance-encoding mcr-9.1 and cephalothin-resis-encoding blaCSA genes and IncFII(pECLA) and IncFIB(pCTU3) plasmids were detected. All strains exhibited CRISPR matrices and four of them two type I-E and I-F matrices. Combined molecular methodologies improve Cronobacter spp. decision-making for health authorities to protect the population.

Estrés académico y estrategias de afrontamiento en estudiantes de carreras de salud de Santiago de Chile / Academic stress and coping strategies in students of health degrees in Santiago de Chile

RESUMEN Fundamento: el stress académico en los estudiantes universitarios puede desencadenar manifestaciones en el orden físico, psicológico y social. Objetivo: determinar el estrés académico y las estrategias de afrontamiento en estudiantes de carreras de salud en Santiago de Chile. Métodos: se realizó un estudio transversal-analítico en una institución de educación superior privada de Santiago de Chile durante el año 2019. Se emplearon los instrumentos de Estrés Académico modificado y la Escala de Afrontamiento del Estrés Académico a la muestra de estudiantes. Se utilizó estadística descriptiva e inferencial, utilizando ANOVA (one-way), test de posthoc de Tukey y Chi2. La significancia se estableció en α<0,05 y los análisis se realizaron con el software STATA 15.0. Resultados: más del 70 % de la muestra se encontraba en un nivel socioeconómico medio-bajo. La situación generadora de estrés predominante estuvo referida a la sobrecarga de trabajo, la cual fue diferente entre los estudiantes de Enfermería (4,1±0,7) en comparación con los de las otras carreras (p=0,032). Ellos presentaron principalmente reacciones físicas ante las situaciones del estrés académico (p=0,024) mientras que los de Terapia Ocupacional tuvieron mayormente reacciones psicológicas (p=0,21). Estos últimos refirieron la revaluación positiva como estrategia de afrontamiento, mientras que el resto de las carreras prefieren la búsqueda de apoyo (p=0.032). Conclusiones: se determinó el estrés académico y sus estrategias de afrontamiento en estudiantes de carreras de salud en Santiago de Chile, quienes identificaron la sobrecarga de trabajo como su génesis sin presentar diferencias entre programas de estudio, pero sí en su afrontamiento.

ABSTRACT Background: academic stress in university students can trigger manifestations in the physical, psychological and social order. Objective: to determine academic stress and coping strategies in students of health degrees in Santiago de Chile. Methods: a cross-sectional-analytical study was carried out in a private higher education institution in Santiago de Chile during 2019. The modified Academic Stress instruments and the Academic Stress Coping Scale were applied to the sample of students. Descriptive and inferential statistics were used, using ANOVA (one-way), Tukey's posthoc test and Chi2. Significance was established at α <0.05 and analyzes were performed with the STATA 15.0 software. Resultados: más del 70 % de la muestra se encontraba en un nivel socioeconómico medio-bajo. La situación generadora de estrés predominante estuvo referida a la sobrecarga de trabajo, la cual fue diferente entre los estudiantes de Enfermería (4,1±0,7) en comparación con los de las otras carreras (p=0,032). Ellos presentaron principalmente reacciones físicas ante las situaciones del estrés académico (p=0,024) mientras que los estudiantes Terapia Ocupacional tuvieron principalmente reacciones psicológicas (p=0,21). Estos últimos refirieron la revaluación positiva como estrategia de afrontamiento, mientras que el resto de las carreras prefieren la búsqueda de apoyo (p=0.032). Conclusiones: se determinó el estrés académico y sus estrategias de afrontamiento en estudiantes de carreras de salud en Santiago de Chile, quienes identificaron la sobrecarga de trabajo como su génesis sin presentar diferencias entre programas de estudio, pero sí en su enfrentamiento.

Presence of Listeria monocytogenes in Ready-to-Eat Artisanal Chilean Foods.

Ready-to-eat (RTE) artisanal foods are very popular, but they can be contaminated by Listeria monocytogenes. The aim was to determine the presence of L. monocytogenes in artisanal RTE foods and evaluate its food safety risk. We analyzed 400 RTE artisanal food samples requiring minimal (fresh products manufactured by a primary producer) or moderate processing (culinary products for sale from the home, restaurants such as small cafés, or on the street). Listeria monocytogenes was isolated according to the ISO 11290-1:2017 standard, detected with VIDAS equipment, and identified by real-time polymerase chain reaction (PCR). A small subset (n = 8) of the strains were further characterized for evaluation. The antibiotic resistance profile was determined by the CLSI methodology, and the virulence genes hlyA, prfA, and inlA were detected by PCR. Genotyping was performed by pulsed-field gel electrophoresis (PFGE). Listeria monocytogenes was detected in 7.5% of RTE artisanal foods. On the basis of food type, positivity in minimally processed artisanal foods was 11.6%, significantly different from moderately processed foods with 6.2% positivity (p > 0.05). All the L. monocytogenes strains (n = 8) amplified the three virulence genes, while six strains exhibited premature stop codons (PMSC) in the inlA gene; two strains were resistant to ampicillin and one strain was resistant to sulfamethoxazole-trimethoprim. Seven strains were 1/2a serotype and one was a 4b strain. The sampled RTE artisanal foods did not meet the microbiological criteria for L. monocytogenes according to the Chilean Food Sanitary Regulations. The presence of virulence factors and antibiotic-resistant strains make the consumption of RTE artisanal foods a risk for the hypersensitive population that consumes them.

Association between high pre-pregnancy body mass index, excess gestational weight gain, and poor breastfeeding with overweight in 3-4-year-old Chilean children.

OBJECTIVE: The aim of this study was to determine the association between prepregnancy body mass, gestational weight gain, and inadequate breastfeeding (BF) with overweight in Chilean children ages 3 to 4 y. METHODS: This was an analytical and cross-sectional study with 560 participants. Age, residence, BF, and weight gain information were collected from child care records. The children's nutritional status (NS) was determined according to the weight-for-height z-score for sex. Bivariate relationships were evaluated by the χ2 test, and a multivariate logistic regression model was applied with the Stata version 15 software at α < 0.05. RESULTS: Prepregnancy NS values were 37% normal and 63% overweight. Excess weight gain occurred in 75% of the mothers. The children's NS was related to the duration of BF (P = 0.002), prepregnancy NS (P = 0.002), and weight gain (P = 0.004). When adjusting the logistic regression model for sex and maternal age, the overweight prepregnancy NS increased up to twice the risk for OW in children (odds ratio [OR], 2; 95% confidence interval [CI], 1.3-4.1), as well as excess weight gain (OR, 2.3; 95% CI, 1.5-5.9), and non-exclusive BF (OR, 2.4; 95% CI, 1.3-4.4). CONCLUSION: Children showing risk factors such as non-exclusive BF, prepregnancy NS with overweight, and excess gestational weight gain faced between 2 and 2.4 times more risk for overweight than children without these factors.