Guillain-Barré syndrome associated with SARS-CoV-2 infection: A case series from 4 Colombian cities during the pandemic.

SARS-CoV-2 infection has been associated with multiple neurological manifestations. One such manifestation, which has been described since the early stages of the COVID-19 pandemic and is relevant for current neurological practice, is Guillain-Barré syndrome (GBS). The literature describes neurotoxic mechanisms of the virus itself and the possible pathways by which it may affect the peripheral nerves in experimental studies; however, we still lack information on the mechanisms causing the immune response that gives rise to GBS in the context of SARS-CoV-2 infection. Colombia is one of the Latin American countries worst affected by the pandemic, with the third-highest number of cases in the region; thus, it is essential to recognise GBS, as this potential postinfectious complication may severely compromise the patient's functional status in the absence of timely diagnosis and treatment. We present a series of 12 cases of GBS associated with SARS-CoV-2 infection from hospitals in 4 different Colombian cities and describe the clinical presentation, laboratory and electrophysiological study findings, and treatment.

En el año 2020 se declaro la pandemia ocasionada por la infección por el virus SARSCoV-2, virus de la familia del coronavirus, adoptándose el nombre de COVID-19 a la enfermedad 1. En Bogotá, Colombia, se confirmó el primer caso de COVID-19 el 6 de marzo de 2020 (2). Los principales síntomas reportados en la infección por SARSCoV-2 son fiebre (43.8% en la admisión y 88.7% durante la hospitalización) y tos (67.8%) (3). Otros síntomas encontrados son fatiga (38.1%), producción de esputo (33.7%) y cefalea (13.6%). Los principales signos neurológicos reportados en los pacientes con infección severa por SARS-Cov-2 son agitación (69%), compromiso en tracto corticoespinal (67%) y delirium (65%) (4). Las principales complicaciones neurológicas descritas asociadas a Covid 19 son: anosmia, disgeusia, encefalopatia, Síndrome de Guillain Barre, complicaciones cerebrovasculares y daño en musculo esquelético (5­8).En el presente articulo se presenta una serie de casos de pacientes con síndrome de Guillain-Barré asociado a infección por SARS-CoV-2. Se recolectaron casos de diferentes instituciones medicas de Colombia.

Gender Differences in Perceived Family Involvement and Perceived Family Control during Emerging Adulthood: A Cross-Country Comparison in Southern Europe.

The aim of the present study was to explore gender differences in perceived parental involvement and perceived psychological and behavioral control during emerging adulthood in two Southern European countries (Spain and Portugal). Data were collected from 491 Portuguese and 552 Spanish undergraduate emerging adults (53.7 % women and 46.3 % men) aged between 18 and 29 years (M = 20.24 and SD = 2.12). Results indicated that women perceived higher levels of parental involvement than men in both countries, and men perceived more behavioral control than women in Portugal. Furthermore, gender was found to moderate the association between perceived parental involvement and perceived psychological and behavioral control differently in each country. Taken together, our findings suggest that gender-differentiated socialization patterns persist during emerging adulthood and that these patterns may be affected by the sociocultural context.

Stability of the Antimicrobial Capacity of Human Milk Against Cronobacter Sakazakii During Handling.

BACKGROUND: Neonatal infections with Cronobacter sakazakii have recently been associated with the consumption of expressed human milk. STUDY AIMS: (1) To evaluate whether human milk has antimicrobial capacity against C. sakazakii and (2) to determine the stability of its capacity when it is subjected to various treatments. METHODS: The antimicrobial capacity of human milk against C. sakazakii was evaluated using an observational, cross-sectional, comparative design. Mature human milk samples (N = 29) were subjected to different treatments. After incubation at 37°C for 72 hr, samples were compared with fresh milk on the stability of their antimicrobial capacity. Two-way analysis of variance (ANOVA) was performed. RESULTS: In fresh milk, counts of C. sakazakii were reduced by 47.26% (SD = 6.74) compared to controls. In treated milk, reductions were: refrigeration at 4°C for 72 hr (M = 33.84, SD = 13.84), freezing at -20°C for 1, 2, and 3 months (M = 40.31, SD = 9.10; M = 35.96, SD = 9.39; M = 26.20, SD = 13.55, respectively), Holder pasteurization (M = 23.56, SD = 15.61), and human milk bank treatment with (M = 14.37, SD = 18.02) and without bovine fortifier (M = 3.70, SD = 23.83). There were significant differences (p < .05) between fresh and treated milk. CONCLUSIONS: Human milk has antimicrobial capacity against C. sakazakii. However, its capacity is negatively influenced by common preservation and hygienization methods. Milk should be stored refrigerated for a maximum of 72 hr or frozen for a short period of time.

Design and application of a 23-gene panel by next-generation sequencing for inherited coagulation bleeding disorders.

INTRODUCTION: Molecular testing of Inherited bleeding coagulation disorders (IBCDs) not only offers confirmation of diagnosis but also aids in genetic counselling, prenatal diagnosis and in certain cases genotype-phenotype correlations are important for predicting the clinical course of the disease and to allow tailor-made follow-up of individuals. Until recently, genotyping has been mainly performed by Sanger sequencing, a technique known to be time consuming and expensive. Currently, next-generation sequencing (NGS) offers a new potential approach that enables the simultaneous investigation of multiple genes at manageable cost. AIM: The aim of this study was to design and to analyse the applicability of a 23-gene NGS panel in the molecular diagnosis of patients with IBCDs. METHODS: A custom target enrichment library was designed to capture 31 genes known to be associated with IBCDs. Probes were generated for 296 targets to cover 86.3 kb regions (all exons and flanking regions) of these genes. Twenty patients with an IBCDs phenotype were studied using NGS technology. RESULTS: In all patients, our NGS approach detected causative mutations. Twenty-one pathogenic variants were found; while most of them were missense (18), three deletions were also identified. Six novel mutations affecting F8, FGA, F11, F10 and VWF genes, and 15 previously reported variants were detected. NGS and Sanger sequencing were 100% concordant. CONCLUSION: Our results demonstrate that this approach could be an accurate, reproducible and reliable tool in the rapid genetic diagnosis of IBCDs.

Determination of lipolytic and proteolytic activities of mycoflora isolated from dry-cured teruel ham.

Fungi play a key role in dry-cured ham production because of their lipolytic and proteolytic activities. In the present study, 74 fungal strains from dry-cured Teruel hams and air chambers were tested for proteolytic and lipolytic activities, with a view to their possible use as starter cultures. Lipolytic activity of fungi was studied against lauric, palmitic, stearic and oleic acids, whereas proteolytic activity was studied against casein and myosin. Of the 74 fungal strains tested, most of them demonstrated lipolytic activity (94.59 %). Lipolytic activity against lauric and oleic acids was stronger than against palmitic and stearic acids. 39 strains (52.70 %) demonstrated proteolytic activity against casein and the 6 highest proteolytic strains were also tested for pork myosin proteolysis. Some strains belonging to Penicillium commune, Penicillium chrysogenum, Penicillium nalgiovense and Cladosporium cladosporioides were selected because of their significant proteolytic and lipolytic activities and could be suitable to use as starters in dry-cured ham.

Mycobiota and toxigenic Penicillium species on two Spanish dry-cured ham manufacturing plants.

The present study reports the natural mycobiota occurring in dry-cured hams, and in particular on the incidence of mycotoxin-producing fungi. A total of 338 fungal colonies were isolated from three stages of production, these being the post-salting, ripening and aging stages in two manufacturing plants. The results show that fungi were more frequently isolated from the aging stage and that the predominant filamentous fungal genus isolated was Penicillium. Seventy-four of the 338 fungal strains were selected for identification at the species level by using morphological criteria and internal transcribed spacers sequencing. Of the 74 fungal strains, 59 were Penicillium strains. Sixteen Penicillium species were identified, with P. commune (24 strains) and P. chrysogenum (13 strains) being the most abundant. The potential ability to produce cyclopiazonic acid (CPA) and ochratoxin A (OTA) was studied by isolating the culture followed by HPLC analysis of these mycotoxins in the culture extracts. The results indicated that 25 (33.7%) of the 74 fungal strains produced CPA. Worth noting is the high percentage of CPA-producing strains of P. commune (66.6%) of which some strains were highly toxigenic. P. polonicum strains were also highly toxigenic. With respect to OTA-producing fungi, a low percentage of fungal strains (9.5%) were able to produce OTA at moderate levels. OTA-producing fungi belonged to different Penicillium species including P. chrysogenum, P. commune, P. polonicum and P. verrucosum. These results indicate that there is a possible risk factor posed by CPA and OTA contamination of dry-cured hams.

Efficacy of low-dose tylvalosin for the control of clostridiosis in broilers and its effect on productive parameters.

The study was carried out under field conditions in a commercial farm, and 1,440 as-hatched Ross-308 broilers were included. Broilers were randomly distributed into 24 experimental 4-m(2) pens (60 broilers/pen). Pens were randomized to the 3 treatment groups: a) tylvalosin 10 mg/kg of live BW during 2 d, b) positive control (tylosin during 2 d), and c) negative control (no treatment). The drugs were provided in the water supply. Mortality, individual BW, and feed intake were assessed. Clostridium presence was assessed in fecal and cecal samples, coccidian oocyst counts were assessed in fecal samples, and bacterial diversity was assessed in ileal content. Live BW at 42 d old was significantly better in the tylvalosin group than in tylosin and no-treatment groups, with tylvalosin-treated broilers reaching 80 to 100 g higher final live weight. Average daily gain results mirrored BW findings. The improvement of feed conversion rate with tylvalosin amounted to 0.13 and to 0.10 versus tylosin and no-treatment, respectively, with mortality being similar in all groups. Significantly reduced sulfite-reducing Clostridium and Clostridium perfringens counts in tylvalosin and tylosin groups versus the no-treatment group were observed in cecum content samples. In conclusion, according to the present study results, tylvalosin, at doses substantially lower than registered for poultry in Europe, has proven effective in controlling the colonization of the cecum by Clostridium ssp. in broilers, improving some productive performances.

Vitamina D: fisiopatología y aplicabilidad clínica en pediatría / Vitamin D: pathophysiology and clinical applicability in paediatrics

La vitamina D se ha relacionado siempre con el metabolismo fosfocálcico, pero se han encontrado receptores de la vitamina D o de sus metabolitos en diferentes células del organismo, lo que indica que puede estar implicada en otros mecanismos fisiológicos. El déficit de vitamina D se ha relacionado con un mayor riesgo de infecciones, enfermedades autoinmunitarias, diabetes, síndrome metabólico, obesidad, asma y ciertas enfermedades neurológicas, como la esquizofrenia. Actualmente existen diferentes técnicas para la determinación del 25 (OH) colecalciferol en sangre y hay discrepancias entre ellas en cuanto a los resultados obtenidos en una misma muestra. Es importante conseguir una uniformización de las técnicas que permitan estudios de investigación comparables. Todavía no se han establecido los valores adecuados de vitamina D en el organismo que aseguren todas sus funciones, pero se considera que no debe ser inferior a 20 ng/ml (50 nml/l), estando en discusión cómo conseguir estos niveles mínimos en toda la población. Dado que la principal fuente de vitamina D es la irradiación solar, hay que buscar estrategias que no entren en contradicción con los mensajes de prevención del cáncer de piel. En los últimos años se han producido variaciones en cuanto las recomendaciones de ingesta de vitamina D que implican a las actividades profilácticas que se realizan en el ámbito de la atención primaria. Este texto revisa la fisiología, las acciones, la determinación de laboratorio, los niveles deseables en el organismo y las recomendaciones de ingesta de la vitamina D, y resalta las numerosas cuestiones que plantean nuevas investigaciones(AU)

Vitamin D has always been associated with calcium -phosphate metabolism, but vitamin D receptors or its metabolites have been found in different body cells, indicating a possible involvement in other physiological mechanisms. Vitamin D deficiency has been associated with an increased risk of infections, autoimmune diseases, diabetes, metabolic syndrome, obesity, asthma and certain neurological diseases such as schizophrenia. Currently there are different techniques for measuring 25 (OH) cholecalciferol in blood, but the results are variable and controversial. It is important to achieve standardization of these techniques to be able to compare the results obtained in different studies. Normal physiological vitamin D levels have not yet been established, but they must be higher than 20 ng/ml (50 nmol/l) in order to perform it physiological function. It is still under discussion on how to achieve these minimum levels. Since the main source of vitamin D is sunlight, we should look for strategies that do not contradict the messages of prevention of skin cancer. In recent years, recommendations for vitamin D intake have changed, involving prophylactic activities carried out in Primary Care. This manuscript reviews the physiology, actions, laboratory determination, desirable levels, and vitamin D intake recommendations, and it highlights many questions raised by new research(AU)

[Vitamin D: pathophysiology and clinical applicability in paediatrics]. / Vitamina D: fisiopatología y aplicabilidad clínica en pediatría.

Vitamin D has always been associated with calcium -phosphate metabolism, but vitamin D receptors or its metabolites have been found in different body cells, indicating a possible involvement in other physiological mechanisms. Vitamin D deficiency has been associated with an increased risk of infections, autoimmune diseases, diabetes, metabolic syndrome, obesity, asthma and certain neurological diseases such as schizophrenia. Currently there are different techniques for measuring 25 (OH) cholecalciferol in blood, but the results are variable and controversial. It is important to achieve standardization of these techniques to be able to compare the results obtained in different studies. Normal physiological vitamin D levels have not yet been established, but they must be higher than 20 ng/ml (50 nmol/l) in order to perform it physiological function. It is still under discussion on how to achieve these minimum levels. Since the main source of vitamin D is sunlight, we should look for strategies that do not contradict the messages of prevention of skin cancer. In recent years, recommendations for vitamin D intake have changed, involving prophylactic activities carried out in Primary Care. This manuscript reviews the physiology, actions, laboratory determination, desirable levels, and vitamin D intake recommendations, and it highlights many questions raised by new research.

Genotypes, exotoxin gene content, and antimicrobial resistance of Staphylococcus aureus strains recovered from foods and food handlers.

Staphylococcal food poisoning, one of the most common food-borne diseases, results from ingestion of one or more staphylococcal enterotoxins (SEs) produced by Staphylococcus aureus in foods. In the present study, 64 S. aureus isolates recovered from foods and food handlers, associated or not associated with food-poisoning outbreaks in Spain, were investigated. They were assigned to 31 strains by spa typing, multilocus sequence typing (MLST), exotoxin gene content, and antimicrobial resistance. The strains belonged to 10 clonal complexes (CCs): CC5 (29.0%), CC30 (25.8%), CC45 (16.1%), CC8, CC15 (two strains each), CC1, CC22, CC25, CC59, and CC121 (one strain each). They contained hemolysin genes (90.3%); lukED (77.4%); exfoliatin genes eta, etd (6.5% each), and etb (3.2%); tst (25.8%); and the following enterotoxin or enterotoxin-like genes or clusters: sea (38.7%), seb (12.9%), sec (16.1%), sed-selj with or without ser (22.9%), selk-selq (6.5%), seh, sell, selp (9.7% each), egc1 (32.3%), and egc2 (48.4%). The number of se and sel genes ranged from zero to 12. All isolates carrying tst, and most isolates with genes encoding classical enterotoxins (SEA, SEB, SEC, and SED), expressed the corresponding toxin(s). Two CC5 isolates from hamburgers (spa type t002, sequence type 5 [ST5]; spa type t2173, ST5) were methicillin resistant and harbored staphylococcal cassette chromosome mec (SCCmec) IVd. Six (19.4%) were mupirocin resistant, and one (spa type t120, ST15) from a food handler carried mupA (MIC, 1,250 µg/ml). Resistance to ampicillin (blaZ) (61.3%), erythromycin (ermA-ermC or ermC) (25.8%), clindamycin (msrA-msrB or msrB) (16.1%), tetracycline (tetK) (3.2%), and amikacin-gentamicin-kanamycin-tobramycin (aphA with aacA plus aphD or aadD) (6.5%) was also observed. The presence of S. aureus strains with an important repertoire of virulence and resistance determinants in the food chain represents a potential health hazard for consumers and merits further observation.

El mesiodens: una patología odontológica que debe tenerse en cuenta / Mesiodens: a dental pathology to consider

Se denomina mesiodens a un diente supernumerario localizado entre los dos incisivos medios superiores o, más raramente, entre los incisivos medios inferiores. Puede producirse en los dos tipos de dentición, pero es mucho más frecuente en la dentición permanente. Clínicamente, se manifiesta por una alteración de la posición o erupción de los incisivos; sólo se manifiesta mediante la erupción del propio mesiodens en un 20-25% de los casos. Su tratamiento suele mejorar el pronóstico de las complicaciones que puede causar, por lo que es conveniente el conocimiento por parte de los pediatras de esta patología para su detección precoz(AU)

A tooth temporary located between the two central superior incisors higher or, more rarely, between the central lower incisors is called mesiodens. It can occur in the two types of teething, but it is much more frequent in the permanent dentition. Clinically it is manifested by an alteration of the position or rash incisors, only manifested through the eruption of mesiodens itself in a 20%-25% of the cases. The treatment may improve the prognosis of the complications that it can cause; the knowledge of the pediatricians of this pathology is appropriate for its early detection(AU)

Virulence and resistance determinants of German Staphylococcus aureus ST398 isolates from nonhuman sources.

A series of 100 Staphylococcus aureus isolates ascribed to sequence type 398 (ST398) and recovered from different sources (healthy carrier and diseased pigs, dust from pig farms, milk, and meat) in Germany were investigated for their virulence and antimicrobial resistance genetic background. Antimicrobial resistance was determined by the disk diffusion method. Virulence and resistance determinants (37 and 31 genes, respectively) were tested by PCR. Only two virulence profiles, including the accessory gene regulator agrI and three or four hemolysin-encoding genes, were detected. In contrast, 33 resistance profiles were distinguished (only 11 were shown by more than one isolate). Fifty-nine isolates were multiresistant (four or more antimicrobial classes), and 98 were methicillin resistant (mecA positive). All of the ST398 isolates showed resistance to tetracycline [encoded by tet(M) alone or together with tet(K) and/or tet(L)]. In addition, 98% were resistant to other antimicrobials, including macrolide-lincosamine-streptogramin B (70%, encoded by ermA, ermB, and ermC, alone or in combination), trimethoprim (65%, mostly due to dfrK and dfrG), kanamycin and gentamicin [29% and 14%, respectively, mainly related to aac(6')-Ie-aph(2″)-Ia and/or ant(4')-Ia but also to aph(3')-IIIa], chloramphenicol (9%, fexA or cfr), quinupristin-dalfopristin (9%), ciprofloxacin (8%), and trimethoprim-sulfamethoxazole (4%). The heterogeneity of the resistance profiles underlines the ability of the ST398 clone to acquire multiple antimicrobial resistance genes. However, the virulence gene content of the tested isolates was low. Continuous surveillance is needed to clarify whether its pathogenicity potential for animals and humans will increase over time.

High heterogeneity within methicillin-resistant Staphylococcus aureus ST398 isolates, defined by Cfr9I macrorestriction-pulsed-field gel electrophoresis profiles and spa and SCCmec types.

During recent years, the animal-associated methicillin-resistant Staphylococcus aureus clone ST398 has extensively been studied. The DNA of these isolates turned out to be refractory to SmaI restriction, and consequently, SmaI is unsuitable for subtyping this clone by standard pulsed-field gel electrophoresis (PFGE). Very recently, ST398 DNA was shown to be digested by Cfr9I, a neoschizomer of SmaI. In the present study, we employed Cfr9I PFGE on 100 German and 5 Dutch ST398 isolates and compared their PFGE profiles, protein A gene variable repeat regions (spa types), and types of the staphylococcal cassette chromosome mec (SCCmec). The isolates (from healthy carrier pigs, clinical samples from pigs, dust from farms, milk, and meat) were assigned to 35 profiles, which were correlated to the SCCmec type. A dendrogram with the Cfr9I patterns assigned all profiles to two clusters. Cluster A grouped nearly all isolates with SCCmec type V, and cluster B comprised all SCCmec type IVa and V* (a type V variant first identified as III) carriers plus one isolate with SCCmec type V. Both clusters also grouped methicillin-susceptible S. aureus isolates. The association of the majority of isolates with SCCmec type V in one large cluster indicated the presence of a successful subclone within the clonal complex CC398 from pigs, which has diversified. In general, the combination of Cfr9I PFGE with spa and SCCmec typing demonstrated the heterogeneity of the series analyzed and can be further used for outbreak investigations and traceability studies of the MRSA ST398 emerging clone.

Aspectos sociofamiliares de la población inmigrante en Cataluña / No disponible

Objetivos: conocer algunas características sociofamiliares de la población inmigranteque llega a las consultas pediátricas de Cataluña.Material y métodos: estudio prospectivo, transversal y multicéntrico. Población: niñosentre 6 meses y 15 años que hacía menos de un año que habían llegado a Cataluñaprocedentes de un país de baja renta (PBR). Variables: sexo, edad, país de procedencia,consanguinidad de los padres, fecha de migración y de primera visita, cuidador principal,familia completa o no, índice de hacinamiento, religión familiar y estudios de lospadres.Resultados: se estudiaron 1.226 pacientes, 659 (53,8%) varones y 567 (46,2%) mujeres.Procedían de 47 países, agrupados en 9 zonas de procedencia. Media de edad de la muestra:7,5 años. Mediana del tiempo entre la llegada a Cataluña y la primera visita al centro de asistenciaprimaria: 1 mes. Cuidador principal: madre o padre en el 96,9%. El 70,9% tenían lafamilia nuclear completa. Consanguinidad entre los padres: 13,2% de los casos. El mayor hacinamiento(índice de hacinamiento [IH] > 2) se da en familias indopakistaníes (49,7%), magrebíes(41,6%) y otros países de Asia (41,8%). Religión familiar: musulmana en el 45,9%de los casos, católica en el 38,8% y otras confesiones cristianas en el 10,5%. No habían ido aescuela: 5,5% de los padres y 16,4% de las madres.Conclusiones: las particularidades culturales y sociales de la población inmigrante hacennecesario su conocimiento para poder ofrecer la mejor asistencia posible a este colectivo(AU)

Objectives: to acquire a better understanding of socio-familiar features of the immigrantpopulation attended in the pediatric offices in Catalonia.Methodology: prospective, transversal and multicentric study. Population: children aged6 months to 15 years, who had arrived to Catalonia less than one year before, and who wereof low income countries’ origin. Variables: sex, age, country of origin, blood relationship betweenthe parents, date of migration and first visit to the medical office, main caregiver, complete/non-complete family, home overcrowding index, religion and parents’ education.Results: one thousand two hundred and twenty-six patients were studied, 659 males(53.8%) and 567 females (46.2%). They had come from 47 countries, which were groupedinto 9 different “origin” groups. Average age of the sample: 7.5 years. Average time betweenarrival to Catalonia and first visit to the doctor: 1 month. Main caregiver: mother or father in96.9% of the cases. In 70.9% of the cases, the family was complete. Parents’ blood relationship:13.2% of the cases. The biggest home overcrowding index (IH > 2) was foundedamong Indian-Pakistan (49.7%), Maghreb (41.6%) and other Asian (41.8%) families. Familyreligion: Muslims (45.9%), Catholic (38.8%) and other Christian doctrines (10.5%). Fivepoint five percent of parents and 16.4% of mothers had never gone to school.Conclusions: in order to provide the best medical assistance to immigrants, it is necessaryto have a good understanding of their cultural and social background(AU)

Clonal complexes and diversity of exotoxin gene profiles in methicillin-resistant and methicillin-susceptible Staphylococcus aureus isolates from patients in a Spanish hospital.

Molecular epidemiology studies have allowed the identification of the methicillin (meticillin)-resistant (MRSA) and methicillin-susceptible (MSSA) clonal complexes (CCs) and clones of Staphylococcus aureus circulating in a Spanish hospital recently. Of 81 isolates tested, 32.1% were MRSA. Most of them carried staphylococcal cassette chromosome mec (SCCmec) IVc (88.5%) and belonged to CC5 (88.5%; multilocus sequence typing types ST125 [mainly associated with spa type t067], ST5, and ST228). A higher diversity was found among MSSA isolates (67.9%). Eighty percent shared the genetic background of major MRSA lineages (CC5 [38.2%; ST125 and ST5], CC30 [25.5%; ST30], CC45 [14.5%; ST45 and ST47], and CC8 [1.8%; ST8]), but CC12, CC15, CC51, and CC59 were also detected. Many exotoxin genes were present in each of the 81 isolates, independent of whether they were involved in sepsis (11 to 22) or other types of infections (13 to 21), and they appeared in 73 combinations. The relevant data are that (i) all isolates were positive for hemolysin and leukotoxin genes (98.8% for lukED and 25.9% for lukPV); (ii) all contained an enterotoxin gene cluster (egc with or without seu), frequently with one or more genes encoding classical enterotoxins; (iii) about half were positive for tst and 95% were positive for exfoliatin-encoding genes (eta, etb, and/or etd); and (iv) the four agr groups were detected, with agrII (55.6%) and agrIII (23.5%) being the most frequent. Taken together, results of the present study suggest a frequent acquisition and/or loss of exotoxin genes, which may be mediated by efficient intralineage transfer of mobile genetic elements and exotoxin genes therein and by eventual breakage of interlineage barriers.

Detection of Salmonella enterica serovar Typhimurium with pUO-StVR2-like virulence-resistance hybrid plasmids in the United Kingdom.

The aim of this study was to investigate the presence in the United Kingdom (UK) of Salmonella enterica serovar Typhimurium isolates carrying pUO-StVR2-like virulence-resistance hybrid plasmids that originated from pSLT. One hundred and fifty ampicillin-resistant isolates of S. Typhimurium, collected in different regions of the UK during 2006, were screened for the presence of bla (OXA-1) carried by an InH-like integron (2000 bp/bla (OXA-1)-aadA1) characteristic of pUO-StVR2. Positive isolates were tested for the presence of a large plasmid that hybridised with probes specific for the bla (OXA-1) and spvC genes, used as resistance and virulence markers of the hybrid plasmid, respectively. Eleven out of the 150 isolates fulfilled both criteria and were assigned to the S. Typhimurium pUO-StVR2 group. Nine were resistant to ampicillin, chloramphenicol, streptomycin/spectinomycin, sulfonamides and tetracycline, encoded by bla (OXA-1), catA1, aadA1-like, sul1 and tet(B), respectively, and carried a pUO-StVR2-like plasmid of ca. 130 kb. Two contained hybrid plasmids of smaller size and lacked resistance(s) to chloramphenicol or chloramphenicol and tetracycline. The eleven isolates, which showed five and six closely related XbaI and BlnI profiles, respectively, were resistant to nitrofurantoin. In conclusion, multidrug-resistant S. Typhimurium isolates of the pUO-StVR2 group, which are endemic in Spain, were also detected in the UK, albeit with a low frequency (7.3%).

Molecular epidemiology of emergent multidrug-resistant Salmonella enterica serotype Typhimurium strains carrying the virulence resistance plasmid pUO-StVR2.

OBJECTIVES: To evaluate the incidence of a distinct multidrug-resistant (MDR) grouping of Salmonella serotype Typhimurium strains carrying the hybrid virulence resistance plasmid pUO-StVR2, and its possible evolution in the region where it was first detected [Principality of Asturias (PA), Spain]. METHODS: pUO-StVR2-containing isolates were tentatively identified by two genetic markers: the bla(OXA-30) gene and the class 1 integron InH:2000 bp/bla(OXA-30)-aadA1a. Positive isolates were examined for resistance profile (RP), plasmid content, virulence profile (VP) and genomic polymorphisms using macrorestriction-PFGE. RESULTS: A total of 182 out of 248 Typhimurium clinical isolates recorded in the PA over 2001-02 were ampicillin-resistant and could be distributed into several MDR groupings. A MDR grouping carrying pUO-StVR2, with a defined RP (AMP/bla(OXA-30), CHL/catA1, [STR-SPT]/[strA/B,aadA1a], SUL/[sul1,sul2], TET/tet(B), qacEDelta1, merA, +/-TMP/dfrA12, and containing InH), was represented by 49 isolates. The VPs of these isolates (24 genes screened) differed from that of the type strain LT2 by the absence of the sopE1 and pef genes. Macrorestriction analysis established six combined XbaI/BlnI PFGE profiles, and supported a clonal relationship among most of the isolates. CONCLUSIONS: During 2001-02, the isolates carrying pUO-StVR2 constituted the second most frequent S. Typhimurium MDR grouping recorded in the PA, preceded only by the pandemic pentaresistant DT104. Polymorphisms on the genomic DNA, different phage types, different plasmid profiles and the detection of trimethoprim resistance in one isolate encoded by an additional plasmid, were consistent with both intra-cluster evolution and horizontal transfer of the hybrid plasmid.

Genetic basis of antimicrobial drug resistance in clinical isolates of Salmonella enterica serotype Hadar from a Spanish region.

The genetic bases of antimicrobial drug resistance (R) of 79 Salmonella enterica serotype Hadar clinical isolates (recovered during 1995-2001 in a Spanish region) was investigated. The isolates showed a limited genomic variation, as demonstrated by PFGE analysis using XbaI (three profiles, S>or=0.77) and BlnI (seven profiles, S>or=0.49; with 95% of the isolates falling into two clusters, S>or=0.75). Thirteen R-profiles, ranging from susceptible to multidrug resistant, were recognized. All susceptible isolates (14%) were recovered before or during 1998, when multidrug resistance (MDR) was still uncommon (20% from 1995-1998). In later years, the percentage of MDR increased considerably (92% in 2001). Resistance to nalidixic acid, tetracycline, streptomycin and ampicillin-cefalotin, encoded by gyrA-Asp87/Asn, tet(A), strA/B, and bla (TEM) genes, respectively, were the most common, appearing together in 38% of the isolates. In all tetracycline- and streptomycin-resistant isolates, strA/B and tet(A) were chromosomally located, whereas bla (TEM) was plasmid-born. Five different bla (TEM) plasmids (pUO-ShR1 to pUO-ShR5, of about 9.4, 23, 30, 45, and 95 kb, respectively) were identified. pUO-ShR3 and pUO-ShR5 harbored additional R-genes: [dfrA1] and [acc(3)IV-strA/B], respectively. pUO-Sh2, pUO-Sh3, pUO-ShR4, and pUO-Sh5 were self-transferable, and the latter could also mobilize pUOShR1. The reported data constitute a useful background for further epidemiological studies of MDR in S. Hadar.

Cytotoxin and pyrogenic toxin superantigen gene profiles of Staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles.

A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands-[lukED, seg, sei, sem, sen, seo +/- seu] with Sabeta-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively-while 11.9% of them carried both islands. Only one contained seb (together with upsilonSabeta-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Sabeta-1, SaPIbov, or both. Five SmaI profiles (S > or = 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (upsilonSabeta-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis.