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Antimicrobial resistance is a global threat, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the most representative drug-resistant pathogens. MRSA spread is increasing due to ...

Introduction The increase in penicillin susceptibility among Staphylococcus aureus (SA-PenS) might have therapeutic relevance. We aimed to study the current situation in our environment. Material and methods Over a 2.5 years period, all SA isolates from bacteraemia were analysed. For all isolates, antimicrobial susceptibility profile, beta-lactam resistance genes (blaZ, mecA) and Panton-Valentine leucocidine encoding-genes were studied. For SA-PenS-blaZnegative isolates, spa-type, MLST and the presence of other resistance genes were studied. Results Among 84 patients with SA bacteraemia (35.7% MRSA and 64.3% MSSA), 77 were analysed; 22.2% of MSSA isolates were PenS and blaZnegative (Pen-MIC ≤0.03 µg/mL) corresponding to 14.3% of the total SA. In MSSA-PenS-blaZnegative isolates, eight spa-types corresponding to seven clonal complexes were detected. Conclusion A high prevalence of MRSA/SA and MSSA-PenS-blaZnegative/MSSA was detected in blood cultures. Pen-MIC ≤0,3 µg/mL corresponded to MSSA-PenS-blaZnegative. This situation raises therapeutic options which should be further evaluated in larger studies and clinical trials.

The increase in penicillin susceptibility among Staphylococcus aureus (SA-PenOver a 2.5 years period, all SA isolates from bacteraemia in one hospital were analysed. For all isolates, antimicrobial susceptibility profile, beta-lactam resistance genes (blaZ, mecA) and Panton-Valentine leucocidine encoding-genes were studied. For SA-PenAmong 84 patients with SA bacteraemia (35.7% MRSA and 64.3% MSSA), 77 were analysed; 22.2% of MSSA isolates were PenA high prevalence of MRSA/SA and MSSA-Pen

Antimicrobial resistance is a global threat, with methicillin-resistant

[EN]: [Introduction]: Methicillin-resistant Staphylococcus aureus (MRSA) is considered a major cause of healthcare-associated (HA) and community-acquired (CA) infections. Considering non-β-lactam susceptibility as a potential marker for mecC-MRSA and CA-MRSA, the aim of this study was to determine the frequency and the associated genetic lineages of non-beta-lactam-antibiotic susceptible MRSA (NBLS-MRSA) strains in a multicenter study in Spain. [Methods]: A collection of 45 NBLS-MRSA strains recovered in the period from January to June 2016 from 12 Spanish hospitals was analyzed. Molecular typing through spa-type characterization, agr group and multi-locus-sequence typing was performed. Methicillin-resistant genes (mecA and mecC) as well as immune evasion cluster (scn-chp-sak-sea-sep, considering scn gene as the marker of IEC system) and Panton-Valentine leukocidin (PVL) genes were determined with PCR/sequencing. [Results]: The NBLS-MRSA phenotype was uncommon in the 12 hospitals analyzed (NBLS-MRSA/MRSA frequency: 0.3%-7.7%). All strains contained the mecA gene (and none contained mecC). Twenty-two different spa-types were detected among NBLS-MRSA strains, with spa-t008/agr-I the most prevalent (27%). The main clonal complexes were (CC/%): CC8/42.2%, CC5/33.3% and CC30/4.4%, with ST8 and ST5 as the main sequence types. The PVL toxin was present in 38% of strains (with spa-types t008, t024, t019, t044, t068, t318 and t3060). The IEC genes were detected in 78% of strains: IEC type-B (n = 17), type-F (n = 16), type-A (n = 1) and type-E (n = 1); 10 MRSA isolates were scn-negative. [Conclusion]: The NBLS-MRSA phenotype is uncommon in the analyzed hospitals; although no mecC-positive strains were detected, it could be a good marker for MRSA PVL-positive isolates (38%), frequently associated with CA-MRSA infections. [ES]: [Introducción]: Staphylococcus aureus resistente a meticilina (SARM) es una de las principales causas de infecciones tanto relacionadas con la asistencia sanitaria como asociadas a la comunidad (AC). Considerando la sensibilidad a antibióticos no-β-lactámicos como marcador potencial de SARM-mecC y SARM-AC, el objetivo de este estudio fue determinar la frecuencia y líneas genéticas de cepas SARM sensibles a antibióticos no-β-lactámicos (SARM-SNBL) en un estudio multicéntrico en España. [Métodos]: Se analizaron 45 cepas SARM-SNBL procedentes de 12 hospitales obtenidas durante enero-junio de 2016. El tipado molecular se realizó mediante caracterización del spa-tipo, grupo agr y multi-locus-sequence typing. Mediante PCR/secuenciación se determinaron los genes: de resistencia a meticilina (mecA y mecC), del sistema de evasión inmune humano (scn-chp-sak-sea-sep, usando scn como marcador del sistema IEC) y de la leucocidina de Panton-Valentine (LPV). [Resultados]: El fenotipo SARM-SNBL fue infrecuente en los 12 hospitales analizados (frecuencia SARM-SNBL/SARM: 0,3-7,7%). Todas las cepas fueron mecA-positivas (ninguna mecC). Se detectaron 22 spa-tipos diferentes, siendo el spa-t008/agr-I el prevalente (27%). Los principales complejos clonales fueron (CC/%): CC8/42,2%, CC5/33,3% y CC30/4,4%, destacando las secuencias tipo ST8 y ST5 como mayoritarias. El 38% de las cepas fue LPV-positiva (spa-tipos t008, t024, t019, t044, t068, t318 y t3060). El 78% de las cepas fue IEC-positivo: tipo-B (n = 17), tipo-F (n = 16), tipo-A (n = 1) y tipo-E (n = 1); 10 aislados fueron scn-negativos. [Conclusión]: El fenotipo SARM-SNBL es poco frecuente en los hospitales analizados; aunque no se detectaron cepas mecC-positivas, este fenotipo puede ser un buen marcador de aislados SARM LPV-positivos, frecuentemente asociados a infecciones por SARM-AC.

Study Group of clinical LA-MRSA, [Background] Tetracycline resistance (TetR) is a marker of livestock-associated MRSA of lineage CC398., [Objectives] To determine the MRSA CC398 prevalence among TetR-MRSA recovered in Spanish hospitals located in regions with different pig-farming densities, and the influence of pig density as a key risk factor for its acquisition., [Methods] TetR-MRSA isolates (n = 232) recovered from clinical and epidemiological samples during January–June 2016 in 20 hospitals in 13 regions with different pig-farming densities were analysed. MRSA CC398 identification, detection of spa types, methicillin resistance genes and immune evasion cluster (IEC) genes were performed by PCR/sequencing. Statistical analyses were performed to establish the relationships between MRSA CC398 prevalence and pig density., [Results] The global MRSA prevalence was 29.7% (6.9% TetR-MRSA/MRSA), with 137 CC398 isolates recovered, representing 4.1% of total MRSA and 59.1% of TetR-MRSA. Among MRSA CC398, 16 different spa types were recorded (t011: 72.3%), and all but two strains were IEC negative. Higher pig-density regions were associated with significant MRSA CC398 increases in hospitals located in adjacent regions (P, [Conclusions] High pig density leads to a significant increase in MRSA CC398 in hospitals in Spain, and its combination with a high human population could help its dissemination. In Spain, the prevalence of the zoonotic CC398 lineage is closely related to pig-farming density; therefore, specific tools could be implemented in order to detect its dissemination., This work was supported by the Sociedad de Enfermedades Infecciosas del Norte (SEINORTE) and by the Agencia Estatal de Investigación (AEI) of Spain (project SAF2016-76571-R) and by the Fondo Europeo de Desarrollo Regional (FEDER). Sara Ceballos and Laura Ruiz-Ripa have predoctoral fellowships of the University of La Rioja, Spain.

Migrating birds have been implicated in pathogen dissemination over long distances. The lack of data on the intestinal microbiota of birds makes these animals a promising path in order to understand their potential role in the transmission of antibiotic-resistant bacteria. This study aimed to investigate the diversity of enterococcal species, and to analyse the antimicrobial-resistant phenotypes/genotypes, as well as the genetic lineages of isolates obtained from faecal and pellet samples of colonial wild birds in Tunisia. Seventy-nine enterococci were recovered from 150 wild birds, after inoculation of samples in Slanetz-Bartley agar, and were identified as E. faecalis (n = 53), E. faecium (n = 19) and E. casseliflavus (n = 7). Antimicrobial susceptibility was tested, and the following rates of resistance were found: tetracycline (46.8%); erythromycin (34.2%); chloramphenicol (8.8%); gentamicin and streptomycin (2.5-3.8%); ciprofloxacin, trimethoprim-sulfamethoxazole and kanamycin (12.7-21%); and ampicillin and linezolid (0%). The tet(M), tet(L), erm(B), erm(C), aac(6')-Ie-aph(2″)-Ia and cat genes were detected in most tetracycline-, erythromycin-, gentamicin- and chloramphenicol-resistant enterococci, respectively. Three vancomycin-resistant E. faecalis isolates were detected, two with the vanA gene (into Tn1546) and one with the vanB2 gene (into Tn5382); these isolates showed different sequence types determined by multi-locus sequence typing (ST9, ST16 and a new ST848). Seven E. casseliflavus isolates harbouring the intrinsic vancomycin resistance mechanism vanC2 were obtained. The gelE, ace, agg, esp and hyl virulence genes were detected among vanA/vanB2 enterococci. This study provides insight into the possible role of wild birds in the spread of certain antimicrobial resistance genes, particularly vanA/vanB2. To the authors' knowledge, this is the first report of vanB2-containing enterococci in Africa.

Plant diseases and pests cause a large loss of world agricultural production. Downy mildew is a major disease in grapevine. Conventional techniques for plant diseases evaluations are time-consuming and require expert personnel. This work investigates novel sensing technologies and artificial intelligence applications for assessing downy mildew in grapevine under laboratory conditions. In our methodology, machine vision is applied to assess downy mildew sporulation, while hyperspectral imaging is used to explore its potential capability towards early detection of this disease. Image analysis applied to RGB leaf disc images is used to estimate downy mildew (Plamopara viticola) severity in grapevine (Vitis vinifera L. cv Tempranillo). A determination coefficient (R2) of 0.76 ** and a root mean square error (RMSE) of 20.53% are observed in the correlation between downy mildew severity by computer vision and expert’s visual assessment. Furthermore, an accuracy of 81% is achieved to detect downy mildew early using hyperspectral images. These results indicate that non-invasive sensing technologies and computer vision can be applied for assessing and quantify sporulation of downy mildew in grapevine leaves. The severity of this key disease is evaluated in grapevine under laboratory conditions. In conclusion, computer vision, hyperspectral imaging and machine learning could be applied for important disease detection in grapevine., Project NoPest (Novel Pesticides for a Sustainable Agriculture)

Diseases and pests cause serious damage in crop production, reducing yield and fruit quality. Their identification is often time-consuming and requires trained personnel. New sensing technologies and artificial intelligence could be used for automatic identification of disease and pest symptoms on grapevine in precision viticulture. The aim of this work was to apply deep learning modelling and computer vision for the detection and differentiation of downy mildew and spider mite symptoms in grapevine leaves under field conditions. RGB images of grapevine canopy leaves with downy mildew symptoms, with spider mite symptoms and without symptoms were taken under field conditions in a commercial vineyard. The images were prepared using computer vision techniques to increase disease visual features. Finally, deep learning was used to train a model capable of differentiating leaf images of the three classes. An accuracy up to 0.94 (F1-score of 0.94) was obtained by classifying leaves with downy mildew, spider mite and without symptoms at the same time, using a hold-out validation. Additionally, accuracies between 0.89 and 0.91 (F1-scores between 0.89 and 0.91) were obtained in the binary classification of the disease and pest, obtaining the best results in differentiating downy mildew from spider mite symptoms. This high accuracy demonstrates the effectiveness of deep learning and computer vision techniques for the classification of grapevine leaf images taken under field conditions, automatically finding complex features capable of differentiating leaves with spider mite symptoms, with downy mildew symptoms and without any. These results prove the potential of these non-invasive techniques in the detection and differentiation of pests and diseases in commercial crop production.

Background Livestock-associated (LA)-CC398-MRSA is closely related to pigs, being unfrequently detected in human invasive infections. CC398-MSSA is emerging in human invasive infections in some countries, but genetic and epidemiological characteristics are still scarcely reported. Objectives To determine the prevalence of Staphylococcus aureus (SA) CC398, both MRSA and MSSA, among blood cultures SA isolates recovered in Spanish hospitals located in regions with different pig-farming densities (PD) and characterize the recovered isolates. Methods One thousand twenty-two SA isolates (761 MSSA, 261 MRSA) recovered from blood cultures during 6–12 months in 17 Spanish hospitals (2018–2019) were studied. CC398 lineage identification, detection of spa-types, and antibiotic resistance, virulence and human immune evasion cluster (IEC) genes were analyzed by PCR/sequencing. Results Forty-four CC398-MSSA isolates (4.3% of SA; 5.8% of MSSA) and 10 CC398-MRSA isolates (1% of SA; 3.8% of MRSA) were detected. Eleven spa-types were found among the CC398-MSSA isolates with t571 and t1451 the most frequent spa-types detected (75%). Most of CC398-MSSA isolates were Immune-Evasion-Cluster (IEC)-positive (88.6%), tetracycline-susceptible (95.5%) and erythromycin/clindamycin–inducible-resistant/erm(T)-positive (75%). No statistical significance was detected when the CC398-MSSA/MSSA rate was correlated to PD (pigs/km2) (p = 0.108). On the contrary, CC398-MRSA isolates were all IEC-negative, predominately spa-t011 (70%), and the CC398-MRSA/MRSA rate was significantly associated to PD (p < 0.005). Conclusion CC398-MSSA is an emerging clade in invasive infections in Spanish hospitals. CC398-MRSA (mostly t011) and CC398-MSSA (mostly t571 and t1451) show important differences, possibly suggesting divergent steps in host-adaptation evolutionary processes. While CC398-MRSA is livestock-associated (lacking IEC-system), CC398-MSSA seems to be mostly livestock-independent, carrying human-adaptation markers.

El síndrome de Wolf-Hirschhorn (SWH) es un síndrome genético neuroevolutivo causado por una deleción del brazo corto del cromosoma 4 (4p16.3). Presentan un fenotipo variable con retraso del crecimiento pre y postnatal, epilepsia, retraso psicomotor, hipotonía, anomalías craneofaciales (facies en “casco griego” y defectos oclusivos), discapacidad intelectual y limitaciones en el uso del lenguaje tanto expresivo como comprensivo. El objetivo de este estudio es describir las habilidades comunicativas y lingüísticas de una niña con fenotipo atenuado de SWH a nivel de desarrollo fonológico, morfológico y léxico. Una niña española de 8;08 años diagnóstica de SWF mediante CGHarray, sin anomalías estructurales orofaciales significativas, salvo laringe muy inmadura. Sus habilidades lingüísticas se evalúan en situación de interacción durante 45 minutos, con pruebas estandarizadas y registro realizado por los padres. Se analiza la frecuencia y la naturaleza de los errores de producción en términos de procesos fonológicos. Los resultados muestran que sus habilidades lingüísticas receptivas equivalen a las de niños de 3;08 años. En situación de interacción produce menos de 50 palabras. No tiene adquirido todo el repertorio fonético español. La frecuencia de los procesos fonológicos es muy alta y característica de niños que se encuentran en los primeros momentos de la etapa verbal del desarrollo fonológico. La participante presenta habilidades fonológicas, morfológicas y léxicas propias de niños de 2-3 años con desarrollo típico. Su perfil lingüístico se ha observado en el retraso del lenguaje y en niños menores de 3;6 años con síndrome de Down. Determinar las características lingüísticas del SWH resulta imprescindible para la intervención logopédica., The Wolf-Hirschhorn syndrome (SWH) is a neurodevelopmental genetic syndrome caused by a deletion of the short arm of chromosome 4 (4p16.3). They present a variable phenotype with pre and postnatal growth delayed, epilepsy, psychomotor delayed, hypotonia, craniofacial anomalies (facies in “Greek helmet” and occlusive defects), intellectual disability and difficulties in the use of both expressive and comprehensive language. The aim of this study is to describe the communicative and linguistic abilities of a girl with an attenuated phenotype of SWH, both at the level of phonological, morphological and lexical development. A Spanish girl aged 8;08 years diagnosed by SWF using CGHarray, without significant structural orofacial anomalies, except very immature larynx participates in this study. Her linguistic abilities are evaluated in interaction situation for 45 minutes, with standardized tests and registration made by the parents. The frequency and nature of production errors in terms of phonological processes are analyzed. The results show that her receptive language skills are equivalent to those of children of 3;08 years old. In situation of interaction produces less than 50 words. She has not acquired all the Spanish phonetic repertoire. The frequency of phonological processes is very high and typical of children who are in the first moments of the verbal stage of phonological development. She presents phonological, morphological and lexical linguistic abilities typical of 2-3-year-old children with typical development. Her linguistic profile has been observed in language delay and in children under 3;6 years with Down syndrome. Determining the linguistic characteristics of SWH is essential for speech therapy intervention.

The in vitro activities of five quinazolinone antibacterials, compounds Q1 to Q5, were tested against 210 strains of methicillin-resistant Staphylococcus aureus (MRSA). The MIC50/MIC90 values (in μg/ml) were as follows: Q1, 0.5/2; Q2, 1/4; Q3, 2/4; Q4, 0.06/0.25; and Q5, 0.125/0.5., The in vitro activities of five quinazolinone antibacterials, compounds Q1 to Q5, were tested against 210 strains of methicillin-resistant Staphylococcus aureus (MRSA). The MIC50/MIC90 values (in μg/ml) were as follows: Q1, 0.5/2; Q2, 1/4; Q3, 2/4; Q4, 0.06/0.25; and Q5, 0.125/0.5. Several strains with high MIC values (from 8 to >32 μg/ml) for some of these compounds exhibited amino acid changes in the penicillin-binding proteins, which are targeted by these antibacterials.

The activities of four oxadiazoles were investigated with 210 methicillin-resistant Staphylococcus aureus (MRSA) strains. MIC50 and MIC90 values of 1 to 2 and 4 μg/ml, respectively, were observed. We also evaluated the activity of oxadiazole ND-421 against other staphylococci and enterococci and in the presence of oxacillin for selected MRSA strains. The MIC for ND-421 is lowered severalfold in combination with oxacillin, as they synergize. The MIC90 of ND-421 against vancomycin-resistant enterococci is ≤1 μg/ml.

The aim of this study was to determine the genetic lineages, and the frequency of antibiotic resistance and virulence determinants in methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) isolates recovered from milk of cows with clinical mastitis. Three hundred milk samples from bovine with clinical mastitis were obtained from 30 dairy farms in different regions of Tunisia. Fifteen of the 300 tested samples contained S. aureus (5%), in three cases were MRSA. Isolates (one/sample) were typed (S. aureus protein A [spa], multilocus sequence typing and accessory gene regulator [agr]). The presence of resistance and virulence genes was analyzed by PCR. The three MRSA isolates contained mecA and blaZ genes (one of them also the msr(A) gene), and carried the enterotoxin gene sen; they were typed as t10381-ST4114 or t267-ST4120, and corresponded to agr type-I. Twelve MSSA isolates were recovered and harbored the blaZ (7 strains) or erm(C) genes (1 strain). The MSSA isolates presented seven different spa-types, associated to new sequence types (STs): t426-ST4118, t267-ST4120, t1773-ST4115, t509-ST4119, t529-ST4117, t2844-ST4113, and t2802-ST4112; most isolates (8/12) were typed as t267/ST4120. All S. aureus isolates were scn-negative, except one MSSA of lineage ST4119 that exhibited the immune evasion cluster type D, and harbored the seg, sei, sem, seo, and seu enterotoxin genes. Four MSSA isolates carried the toxic shock syndrome toxin 1 gene (tst). S. aureus (including MRSA) is an important cause of bovine mastitis, showing isolates with high genetic diversity and high content in virulence genes.

Calanoides carinatus sensu lato (s.l.) (Krøyer, 1848) is known to occur in the North and South Atlantic, Indian, and tropical western Pacific Oceans, and the Mediterranean Sea. In highly productive areas, this species may dominate the copepod biomass and occupy a pivotal position in the pelagic food webs that support commercial fisheries. Despite the species´ ecological importance, its taxonomic status and biogeographic range are not certain. An international collaborative projectwas carried out to examine the molecular phylogeography and population genetics of C. carinatus of the Atlantic Ocean. A total of 55 specimenswere identified from13 samples collected from neritic populations of the SW (23°S?47°S), SE (16°S?34°S) and NE (11°N?44°N) Atlantic Ocean. A 708 base-pair (bp) region of the mitochondrial cytochrome c oxidase subunit I (mtCOI) gene was amplified. DNA sequences were trimmed to a final aligned length of 484 bp for analysis. A parsimony haplotype network (constructed with TCS Ver. 1.2.1) had no shared haplotypes between the SE and SW Atlantic samples. In contrast, SE and NE Atlantic samples had broadly shared haplotypes, although several individuals from the NE Atlantic had very divergent haplotypes. Based on all samples analyzed, neutrality tests were both positive and significant (Tajima´s D=2.374, p b 0.05; Fu´s FS=29.378, p b 0.001) and haplotype diversity (Hd) was 0.81. NE and SE Atlantic samples were pooled for analysis (based on average pairwise Tamura-Nei distance between C. carinatus s.l. individuals = 0.024). Between SW and NE/SE Atlantic populations, average pairwise distance = 0.556 and φST = 0.956 (p b 0.0001). Phylogeographic analysis of mtCOI sequence variation indicates that C. carinatus s.l. comprises two genetically divergent and geographically distinct species. Since the type locality of the species is in Brazilian waters, we consider the SW Atlantic type to represent C. carinatus sensu stricto (s.s.), with a biogeographical range restricted to the SW Atlantic Ocean. Further, we consider the NE/SE Atlantic type to be an undescribed, cryptic sibling species. Fil: Viñas, Maria Delia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Subsede Instituto Nacional de Investigación y Desarrollo Pesquero; Argentina Fil: Blanco Bercial, Leocadio. University of Connecticut; Estados Unidos Fil: Bucklin, Ann. University of Connecticut; Estados Unidos Fil: Verheye, Hans. Oceans and Coastal Research; Sudáfrica Fil: Bersano, José Guilerme F.. Universidade Federal do Paraná; Brasil Fil: Ceballos, Sara. Instituto Español de Oceanografía; España

Introduction Staphylococcus aureus and Staphylococcus pseudintermedius are highly important due to their capacity for producing diseases in humans and animals, respectively. The aim of the study was to investigate and characterize the coagulase positive Staphylococcus (CoPS) carriage in a Primary Healthcare Center population. Methods Nasal swabs were obtained from 281 non-infectious patients. The CoPS isolates recovered were typed, and their resistance phenotype and genotype, as well as their virulence profiles, were analyzed. Results CoPS isolates were recovered from 56/281 patients (19.9%). Fifty-five were S. aureus (19.6%), 54 were methicillin susceptible (MSSA) and one was methicillin resistant (MRSA). The remaining isolate was S. pseudintermedius (0.4%). A high diversity of spa-types (n = 40) was detected, with 6 of them being new ones. The multi-locus-sequence-typing of 13 MSSA and one MRSA selected isolates was performed and the STs detected were: ST8, ST15, ST30, ST34, ST121, ST146, ST398, ST554, ST942, ST2499, and ST2500 (the last two STs being new). One MSSA isolate was typed as t1197-ST398-(Clonal complex)CC398. The MRSA isolate was typed as t002-ST146-CC5-SCCmec-IVc, and exhibited a multiresistance phenotype. The detected resistances were: penicillin (76%), macrolides (7%), tetracycline (7%), trimethoprim-sulfamethoxazole (7%), quinolones (7%), and lincosamides (5%). Five isolates contained lukF/lukS-PV genes, 17 tst gene, one eta gene, and two etb gene. The S. pseudintermedius isolate presented a new spa-type (t57) (belonging to a new ST180) and the genes lukS/F-I, siet, se-int, and expB. Conclusions A high genetic diversity of S. aureus was detected. Mention must be made of the identification of MSSA CC398 and S. pseudintermedius isolates in two patients, one of them with animal contact. The detection of the genes lukF/lukS-PV and tst should be noted.

Tetracycline-resistance (Tet(R)) has been postulated as a marker of the livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) lineage CC398.to determine the spa-types and assigned MLST clonal complexes (CCs) among all 98 MRSA-Tet(R) strains recovered during 2011-2012 (from different patients) in a Spanish Hospital, analyzing the possible correlation with livestock-contact of the patients. All 98 strains were assigned to 9 CCs: CC398 (60.2%), CC1 (19.4%), CC5 (12.2%), and other CCs (8.2%). The 98 patients were classified into three groups: (A) contact with livestock-animals (n=25); (B) no-contact with livestock-animals (n=42); (C) no information about animal contact (n=31). A significant higher percentage of CC398 strains was obtained in group A (76%) than in group B (50%) (p0.05), being the percentage in group C of 61.3%. Most of MRSA-Tet(R)-CC398 strains presented a multi-resistance phenotype, including erythromycin, clindamycin, and ciprofloxacin, and the most prevalent detected genes were tet(M) and erm(C). Three strains presented the phenotype macrolide-susceptibility/lincosamide-resistance and contained the vga(A) gene. MRSA-CC1 strains showed higher percentages of erythromycin/clindamycin resistance (95%/89%) than MRSA-CC398 strains (58%/63%), and this resistance was usually mediated by erm(C) gene. Most of MRSA-CC5 strains showed resistance to ciprofloxacin, tobramycin/kanamycin and erythromycin. None of the strains presented the genes lukF/lukS-PV, tsst-1, eta, etb or etd. All MRSA-CC398 strains lacked the genes of the immune-evasion-cluster, but MRSA-CC1 strains carried these genes (type E). In conclusion, although MRSA CC398 is detected in a significant higher proportion in patients with livestock-contact; its detection in people without this type of contact also indicates its capacity for human-to-human transmission.

Sexual selection is potentially important in marine zooplankton, presumably the most abundant metazoans on earth, but it has never been documented. We examine the conditions for sexual selection through mate choice and describe mating preferences in relation to size in a marine zooplankter, the pelagic copepod Acartia tonsa. Males produce spermatophores at a rate (~1 day(-1)) much lower than known female encounter rates for most of the year and the decision to mate a particular female thus implies lost future opportunities. Female egg production increases with female size, and males mating larger females therefore sire more offspring per mating event. Similarly, females encounter males more frequently than they need to mate. Large males produce larger spermatophores than small males and the offspring production of female increases with the size of the spermatophore she receives. Additionally, large spermatophores allow females to fertilize eggs for a longer period. Thus, mating with large males reduces the female's need for frequent matings and she may sire sons that produce more offspring because size is heritable in copepods. Finally, we show that both males and females mate preferentially with large partners. This is the first demonstration of sexual selection by mate choice in a planktonic organism.

The optimal allocation of resources to repair vs. reproduction in an organism may depend on the magnitude and pattern of the external mortality it is experiencing, which, in turn, may depend on its feeding and mate-finding behavior. Thus, the fundamental activities of an organism, i.e., to feed, to survive, and to reproduce, are interrelated through trade-offs. Here, we use small planktonic copepods to examine how adult longevity and ageing patterns in a protected laboratory environment relate to the feeding mode (active searching vs. passive ambush feeding), mate-finding behavior, and spawning mode of the species. We show that average adult longevity varies between species by an order of magnitude and is independent of body size. Ambush feeders that carry their eggs have longer average life spans and experience higher mortality later in life relative to active feeders that broadcast their eggs. Males generally have shorter life spans and experience higher mortality earlier in life than females, and this difference may be accentuated in species where dangerous mate-finding is male biased. We finally show a trade-off between longevity and fecundity, with ambush feeders producing eggs at a rate five to 10 times lower than the active feeders, consistent with predictions from optimal resource allocation theory.

The detection of vancomycin-resistant-enterococci (VRE) among wild animals represents a worrisome public health concern. The objectives of the study were to determine the possible presence of VRE in faecal samples of wild small mammals in Spain, to characterize the vancomycin resistance mechanisms and genetic lineages of recovered isolates and to know the diversity of enterococcal species in these animals. A total of 155 faecal samples from small mammals were inoculated in Slanetz-Bartley agar supplemented or not with vancomycin (Van-SB/SB plates). The antimicrobial susceptibility profile to 12 antimicrobials and the presence of 20 antimicrobial resistance genes was analyzed. The structure of Tn. 1546 and the presence of gelE, cylA, asa, esp and hyl genes was studied. Multilocus-sequence-typing (MLST) technique was also performed. VRE isolates were recovered in Van-SB plates in 11 samples. Two samples contained vanB2-positive E. faecalis isolates of lineage ST6, which showed a multiresistance phenotype and harboured the virulence genes gelE and asa. One sample contained a vancomycin-resistant E. faecium isolate of the new lineage ST915, with the vanA gene included into Tn. 1546 (truncated with IS. 1542 and IS. 1216 elements). The vanB2 and vanA isolates were obtained from Rattus rattus. The remaining eight VRE-positive samples contained species with intrinsic vancomycin-resistance mechanisms: E. casseliflavus (n= 5) and E. gallinarum (n= 3). One hundred and forty-seven vancomycin-susceptible-enterococcal isolates were obtained in SB plates, and E. faecalis and E. faecium were the most frequent detected species. This is the first report of vanB2-containing enterococci in wild animals., This work was supported by Projects SAF2012-35474 and CGL2011-30274 from the Ministerio de Economía y Competitividad (MINECO) of Spain and the Fondo Europeo de Desarrollo Regional (FEDER). C. Lozano has a contract associated with Project SAF2012-35474. F. Ruiz-Fons acknowledges funding from MINECO through a Ramon y Cajal contract reference RYC-2013-12528.

The reproductive activity and the physiological state of the calanoid copepods Calanus helgolandicus and Calanoides carinatus were investigated off the coast of NW Spain during autumn to evaluate the effect of short food resources on both populations. Phytoplankton biomass was low, and neither phytoplankton size distribution nor composition was suitable to support high reproductive rates. Accordingly, egg production rates (EPR) were much lower than maximum rates for both species, pointing to food limitation. The reproductive index (RI), which represents the proportion of females with mature gonads, was b50% at each of the three zones into which the sampling area was divided (coast, shelf and ocean). Potential recruitment rates were very low except at some nearshore stations, where the highest concentrations of chlorophyll-a (Chl-a), diatoms, dinoflagellates and large cells were found. EPR of C. helgolandicus and C. carinatus were correlated with phytoplankton biomass and unaffected by temperature. Phytoplankton carbon ingestion explained ca. 50% of the variability in EPR for both species. At most of the stations, herbivory was insufficient to cover the carbon requirements for reproduction and respiration, so females probably fed on heterotrophic prey to meet their demands. However, given the low fecundity observed, this omnivorous diet did not seem to be optimum for reproduction, and a severe food limitation is thus suggested. Furthermore, the high C/N values measured point to a notable lipid storage, but given the low EPR found, lipid reserves were probably invested into female maintenance rather than into gonad maturation. C. helgolandicus and C. carinatus populations did not mirror phytoplankton biomass distribution, but they correlated well when considering only copepodites V (CV). The CV could be preparing for the overwintering, storing lipid reserves to ensure a successful diapause, and they could also be advected by the poleward current detected during the study. Females showed a diel feeding rhythm, with highest ingestion rates during night. From our results, it follows that C. helgolandicus and C. carinatus females did not perform diel vertical migrations. We suggest that this behaviour is likely due to the food-limiting conditions, which make it more advantageous to remain at the surface during daytime.

The reproductive activity of 2 calanoid copepods, Calanus helgolandicus and Calanoides carinatus, under post-bloom conditions was investigated during a cruise in the central Cantabrian Sea (SW Bay of Biscay) in May 2000. Several mesoscale structures, such as an upwelling of rich waters associated with the topography of one submarine canyon, were observed on the continental shelf. Consequently, a high biomass of large phytoplankton was found in coastal and mid-shelf areas. These unexpected productive conditions after the spring bloom supplied copepods with favourable food resources. Accordingly, females showed mature gonads, high percentages of spawning and high egg production rates (EPRs). A surface saline current was detected flowing parallel to the coast along the slope. A microbial food web prevailed there and, unlike the coast and mid-shelf areas, low chlorophyll concentrations and a high proportion of small phytoplankton were found. The microbial food web did not seem to be suitable to support copepod production; thus, only a few females had mature gonads and fecundity was low. The number of eggs produced increased as herbivorous feed- ing increased, but feeding on phytoplankton did not cover the carbon requirements at all stations, and a mixed diet was suggested. The relationship between EPR and diatom concentration showed a saturation response for both species, but interestingly the saturation concentration was low and fecundity values were sub-maximal. Females had low lipid levels and low C:N ratios, indicating that egg production could be entirely fuelled by freshly ingested food. The low lipid reserves of females could also help to explain the sub-maximal EPR of C. helgolandicus. Nevertheless, low lipid storage was not a constraint for C. carinatus because it is well known that this species uses only freshly ingested food to support egg production. The implications of this extra productive time at the end of spring on the annual recruitment of C. helgolandicus and C. carinatus are discussed.

The seasonal reproductive dynamics of the calanoid copepods Calanus helgolandicus and Calanoides carinatus and the main driving factors that control their reproductive activity were evaluated through a three-year study performed at two shelf stations in the Cantabrian Sea (SW Bay of Biscay). As a general pattern, the seasonal dynamics showed two peaks of reproduction for both species: the highest peak was associated with the spring phytoplankton bloom, whereas a second and less intense peak coincided with the autumn bloom. Between these two blooms, copepod recruitment showed low rates, and the lowest reproductive activity was recorded in winter. Gonads started to mature in pre-bloom conditions, probably fuelled by stored lipids and concurrent feeding, but the highest RIs (Reproductive Index) and EPRs (egg production rates) were found during the spring blooms. Accordingly feeding seems to be crucial for the main recruitment event for both species. Temperature, Chlorophyll-a (Chl-a) concentration and diatom abundance explained part of the variability found in reproductive activity, with the phytoplankton biomass as the most relevant environmental factor. Temperature probably had an indirect effect on fecundity; temperature and female size were inversely correlated, and large females produced larger clutch sizes. The food assemblage seemed to be more adequate for egg hatching success (HS) than for EPR, because HS was high most of the time, but EPRs were often lower than maximum production rates, pointing to a food limitation. However, the nauplius production rates compared well with those in other areas with higher phytoplankton productivity. Interestingly, a trade-off between number of eggs produced and their viability was found; small clutch sizes with high viability were produced during inadequate food conditions. This strategy maximizes naupliar survival under short food resources by investing in egg quality rather than in quantity. The annual cycle and reproductive dynamics of C. carinatus , traditionally described for African upwelling systems, were compared with that of C. helgolandicus , a resident species in the Bay of Biscay. The high similarity between the species suggests that C. carinatus has developed a stable population in the Cantabrian Sea, which denotes a high plasticity of this species.

Mesozooplankton size-fractionated biomass, feeding, and metabolic rates were investigated during a cruise conducted off NW Spain in August 1998. Based on different hydrographic conditions observed throughout the study area (upwelling events nearshore and stratified waters offshore), the sampling area was divided into coastal and oceanic zones. Both phytoplankton and mesozooplankton biomass were higher in the coastal upwelling than in the offshore zone. Size structure analysis shows that the small fraction (200e500 mm) of mesozooplankton was more important than the larger fractions (500e1000 mm and O1000 mm) in the open ocean in terms of both biomass and grazing activity. The relationship between copepod ingestion rate and the concentration of chlorophyll a O5 mm resembled a type II functional response. Saturation occurred at the coastal stations, where copepod gut contents were highest. The grazing impact on both phytoplankton biomass and primary production seemed to be higher offshore (5.7 and 12.9%, respectively) than in upwelled waters (5.2 and 5.2%), although the differences were not statistically significant. Phytoplankton ingestion was enough to fulfil the basal metabolism of mesozooplankton in the upwelling area (102.1% of the minimum carbon requirements met) but not in the offshore (45.9%). Feeding activity and metabolic measurements on mesozooplankton point to a herbivorous food web in the coastal upwelling zone and a multivorous food web in the stratified open oceanic waters. The amount of metabolic end products released by mesozooplankton was higher in the coastal zone, but mesozooplankton played a more important role as nutrient regenerators in the multivorous food web (30.3 and 21.7% of the nitrogen and phosphorus phytoplankton demand accounted for ammonium and phosphate excretion, respectively) than in the herbivorous one (3.8% for ammonium and 2.6% for phosphate). 2004 Elsevier Ltd. All rights reserved.

Mesozooplankton distribution, metabolism and feeding were investigated in the slope water oceanic anticyclonic eddy (SWODDY) AE6 during the cruise Gigovi-0898, which was conducted in August 1998 in the Bay of Biscay. According to the distribution of isotherms at 200 m depth, the sampling area was divided into three zones: SWODDY centre (C), edge (E) and outside the SWODDY (O). Multivariate analysis identified four different zooplankton assemblages. Such separation was closely related to the structure of the SWODDY, with each grouping associated to a particular zone of this mesoscale structure. Thus, two of the groups were located at the SWODDY centre and outside (groups C and O, respectively), whereas the two remaining station groupings (TWE and TNS) were situated at the SWODDY margin. The assemblages C and O differed clearly from each other in their taxonomic composition. In contrast, TWE and TNS were separated because of the differences observed in numerical abundance rather than by a different composition. Some taxa were found exclusively outside the SWODDY, while others were confined to the centre, suggesting a large degree of isolation of the SWODDY with respect to the surrounding waters. Phytoplankton and mesozooplankton biomass followed a similar pattern, with highest values in the SWODDY centre, although the differences between zones were significant only for phytoplankton. The highest grazing impact for both phytoplankton standing stock (3.4%) and primary production (12.7%) was observed outside the SWODDY. Ammonium released by mesozooplankton accounted for between 25.3% (outside the SWODDY) and 44.4% (SWODDY centre) of total phytoplankton nitrogen demands. The percentage of total phosphorus required to be fulfilled by phosphate excretion ranged from 14.0% at the edge to 17.5% outside. Mesozooplankton underwent intense vertical migrations, especially at the SWODDY edge. The role of the mesozooplankton in the biological pump releasing metabolic end-products below 200 m at night time is proposed as one of the main sources of downward export of biogenic material in these systems.

Four diatoms, Thalassiosira rotula (THA), Thalassiosira weissflogii (TWEI), Skeletonema costatum (SKE) and Phaeodactylum tricornutum (PHA), and one dinoflagellate Prorocentrum minimum (PRO, control diet), were tested during the course of 14-day laboratory experiments examining the effects of these algae on the fecundity and egg viability in the copepod Temora stylifera . All algae were provided at the same mean carbon concentrations of 0.98 μg C ml −1 to normalize the effects due to differences in diatom size and nutrient concentrations, at least with regards to carbon, so that variations in egg production and hatching viability were presumably due to other chemical constituents of the cells. Our results show that only PRO supported high reproductive success, with stable egg production rates and egg viability close to 100%. By contrast, all four diatoms negatively impacted egg production and hatching success, with varying degrees of suppression. The two diets SKE and PHA induced dramatic effects on reproductive success; SKE was the worst diet with naupliar production ceasing altogether within only 3 days. With PHA, fecundity never reached zero, but at the end of the experiments, initial egg production rates had dropped to 6.6% of initial rates, and viability had declined to zero within 7 days. The two Thalassiosira species mainly affected egg viability, so that after 14 days of feeding, a decrease of 90% had occurred. However, egg production rates were high and were reduced only by 15–24% towards the end of the experiments. In terms of potential recruitment rates, even though PRO induced high mean naupliar production, a slight decrease was observed during the first 5 days, likely due to the time required for T. stylifera to acclimate to PRO. After day 5, potential recruitment rate was higher, comparable to in situ values. THA and TWEI inhibited potential recruitment up to 90–100%, but these effects were not evident over a short-term period, denoting the importance of introducing the time factor when evaluating diatom effects on copepod production. With the other two diatom diets, SKE and PHA, recruitment was zero after 2 and 6 days, respectively. Experiments testing the effects of unsaturated aldehydes in THA (2-trans-4-trans-decadienal) and SKE (2-trans-4-trans-octadienal and 2-trans-4-trans-heptadienal) on the hatching viability of T. stylifera indicate that diatom-derived aldehydes may differ in terms of biological activity. Of the three molecules tested, decadienal had somewhat stronger effects on hatching success, with total blockage occurring at concentrations of 2.0 μg ml −1 ; the concentrations to induce total blockage by octadienal and heptadienal were somewhat higher and ≥2.5 μg ml −1 . We suggest that the greater biological activity of SKE in terms of cell blockage may thus be due to the presence of antimitotic compounds other than these unsaturated aldehydes.

14 pages, 8 figures, 2 tables, Reproduction in planktonic animals depends on numerous biotic and abiotic factors. One of them is predation pressure, which can have both direct consumptive effects on population density and sex ratio, and non-consumptive effects, for example on mating and migration behaviour. In copepods, predator vulnerability depends on their sex, motility pattern and mating behaviour. Therefore, copepods can be affected at multiple stages during the mating process. We investigated the reproductive dynamics of the estuarine copepod Eurytemora affinis in the presence and absence of its predator the mysid Neomysis integer in a mesocosm experiment. We found that the proportion of ovigerous females decreased in the presence of predators. This shift was not caused by differential predation as the absolute number of females was unaffected by mysid presence. Presence of predators reduced the ratio of males to non-ovigerous females, but not by predation of males. Our combined results suggest that the shift from ovigerous to non-ovigerous females under the presence of predators was caused by either actively delayed egg production or by shedding of egg sacs. Nauplii production was initially suppressed in the predation treatment, but increased towards the end of the experiment. The proportion of fertilized females was similar in both treatments, but constantly fell behind model predictions using a random mating model. Our results highlight the importance of non-consumptive effects of predators on copepod reproduction and hence on population dynamics. © 2014 Springer-Verlag Berlin Heidelberg, The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under Grant Agreement No. 228224, MESOAQUA (Grant Number: MESOAQUA 16. COPEPODSEX.HEUSCHELE). J. H. was funded by a Deutsche Forschungsgemeinschaft Research Fellowship (grant number HE 6050/1-1)

We review current knowledge and understanding of the biology and ecology of the calanoid copepod Calanus helgolandicus in European waters, as well as provide a collaborative synthesis of data from 18 laboratories and 26 sampling stations in areas distributed from the northern North Sea to the Aegean and Levantine Seas. This network of zooplankton time-series stations has enabled us to collect and synthesise seasonal and multi-annual data on abundance, body size, fecundity, hatching success and vertical distribution of C. helgolandicus. An aim was to enable comparison with its congener Calanus finmarchicus, which has been studied intensively as a key component of European and north east Atlantic marine ecosystems. C. finmarchicus is known to over-winter at depth, whereas the life-cycle of C. helgolandicus is less well understood. Overwintering populations of C. helgolandicus have been observed off the Atlantic coast between 400 and 800 m, while in the Mediterranean there is evidence of significant deep-water populations at depths as great as 4200 m. The biogeographical distribution of C. helgolandicus in European coastal waters covers a wide range of habitats, from open ocean to coastal environments, and its contribution to mesozooplankton biomass ranges from 6% to 93%. Highest abundances were recorded in the Adriatic and off the west coast of Spain. C. helgolandicus is generally found in 9–20 C water, with maximum abundances from 13–17 C. In contrast, C. finmarchicus is found in cooler water between 0 and 15 C, with peak abundances from 0 to 9 C. As water has warmed in the North Atlantic over recent decades, the range of C. helgolandicus and its abundance on the fringes of its expanding range have increased. This review will facilitate development of population models of C. helgolandicus. This will not only help answer remaining questions but will improve our ability to forecast future changes, in response to a warming climate, in the abundance and distribution of this important species. Versión del editor

Reproductive activity and production of the calanoid copepods Calanus helgolandicus and Calanoides carinatus were measured during a summer upwelling event off the coast of NW Spain. The upwelling pattern affected the distribution and fecundity of both species in the study area. The demographic composition of both populations and the stage of gonad maturation (e.g. the high abundance of fertilised females with mature ova) indicated active reproduction. C. carinatus, a highly fecund species associated with the African upwelling zones and considered as an “upwelling specialist”, showed low production rates (overall means of 15 eggs female−1 day−1 and 3% body C day−1), despite the fact that the food conditions (high phytoplankton biomass dominated by diatoms) seemed to be optimal for this species. By contrast, C. helgolandicus, a temperate species that shows a strong link between spring phytoplankton blooms and reproduction time, seems to be flexible enough to take full advantage of shorter-term, enhanced feeding conditions associated with the pulsed nature of the summer coastal upwelling. Both the egg and carbon-specific production rates attained by this species (overall means of 26 eggs female−1 day−1 and 12% body C day−1) were similar to values reported for a spring bloom situation. This high production would imply a long spring–summer recruitment event of C. helgolandicus in these waters. For both species the stage of gonad maturation was significantly correlated with their egg production rates and likely influenced by the food conditions; a species-specific nutritional requirement for final oogenesis is suggested. The carbon condition factor (carbon weight/prosome volume) of C. carinatus females was higher than that of C. helgolandicus, suggesting differential use of the carbon ingested; C. helgolandicus seems to use all ingested carbon to produce eggs at a high rates, whereas C. carinatus seems to store part of the ingested carbon as lipid reserves to ensure female survival and to support production during subsequent unfavourable food conditions.