Carmelo Ortega

Correlation of Borrelia burgdorferi Sensu Lato Prevalence in Questing Ixodes ricinus Ticks with Specific Abiotic Traits in the Western Palearctic

ABSTRACT This meta-analysis of reports examining ticks throughout the Western Palearctic region indicates a distinct geographic pattern for Borrelia burgdorferi sensu lato prevalence in questing nymphal Ixodes ricinus ticks. The greatest prevalence was reported between the 5°E and 25°E longitudes based on an analysis of 123 collection points with 37,940 nymphal tick specimens (87.43% of total nymphs; 56.35% of total ticks in the set of reports over the target area). Climatic traits, such as temperature and vegetation stress, and their seasonality correlated with Borrelia prevalence in questing ticks. The greatest prevalence was associated with mild winter, high summer, and low seasonal amplitude of temperatures within the range of the tick vector, higher vegetation indices in the May-June period, and well-connected vegetation patches below a threshold at which rates suddenly drop. Classification of the target territory using a qualitative risk index derived from the abiotic variables produced an indicator of the probability of finding infected ticks in the Western Palearctic region. No specific temporal trends were detected in the reported prevalence. The ranges of the different B. burgdorferi sensu lato genospecies showed a pattern of high biodiversity between 4°W and 20°E, partially overlapping the area of highest prevalence in ticks. Borrelia afzelii and Borrelia garinii are the dominant species in central Europe (east of ∼25°E), but B. garinii may appear alone at southern latitudes and Borrelia lusitaniae is the main indicator species for meridional territories.

Identification of novel vga(A)-carrying plasmids and a Tn5406-like transposon in meticillin-resistant Staphylococcus aureus and Staphylococcus epidermidis of human and animal origin.

Nine staphylococcal strains of human and animal origin with a lincomycin-resistant/erythromycin-susceptible phenotype and carrying vga genes were characterised to determine the genetic elements involved in the dissemination of these uncommon resistance genes. These strains were typed by multilocus sequence typing (MLST), staphylococcal cassette chromosome mec (SCCmec) and/or spa typing. Antimicrobial susceptibility was studied by disk diffusion and agar dilution methods. Presence of the genes lnu(A), lnu(B), vga(A), vga(A)v, vga(B), vga(C), vga(E), lsa(B) and cfr was studied by PCR. Transformation experiments were carried out in all strains, and the plasmid or chromosomal gene location was determined by Southern blot analysis. Genetic environments of the vga genes were analysed by PCR mapping or inverse PCR and sequencing. Five meticillin-resistant Staphylococcus aureus (MRSA) ST398 strains and three Staphylococcus epidermidis strains harboured the gene vga(A), and one MRSA-ST8 strain contained the gene vga(A)v. One MRSA-ST398 strain, which also contained the gene lnu(A), showed the highest minimum inhibitory concentration (MIC) to lincomycin. The vga(A)v-positive strain presented lower MIC values than the vga(A)-positive strains. Presence of the pVGA plasmid was confirmed in two MRSA-ST398 strains. Four novel vga(A)-carrying plasmids were detected: pUR2355 (in two MRSA and one meticillin-susceptible S. epidermidis); pUR4128 (one MRSA); pUR3036 [one meticillin-resistant S. epidermidis (MRSE)]; and pUR3937 (one MRSE). The plasmid pUR4128 was very similar to pUR2355. Plasmids pUR3036 and pUR3937 were related and were very similar to plasmid pSE-12228-06. The gene vga(A)v was located in a transposon analogous to Tn5406. Therefore, four novel vga(A)-carrying plasmids and a variant of Tn5406 were identified in this study.

Wild Birds, Frequent Carriers of Extended-Spectrum β-Lactamase (ESBL) Producing Escherichia coli of CTX-M and SHV-12 Types

To get a better insight into the role of birds as reservoirs of extended-spectrum β-lactamase (ESBL) and plasmidic AmpC β-lactamase (pAmpC) Escherichia coli producers, 100 fecal samples belonging to 15 different wild avian species from Northern Spain were analyzed. Cefotaxime-resistant (CTXR) E. coli isolates were identified in 16 of the 100 tested birds, which corresponded to 9 animal species (Gyps fulvus—griffon vulture, Larus michahellis—yellow-legged gull, Milvus migrans—black kite, Milvus milvus—red kite, Ciconia ciconia—white stork, Sturnus unicolor—spotless starling, Aquila chrysaetos—golden eagle, Cuculus canorus—common cuckoo, Tyto alba—barn owl). Fifteen isolates harbored ESBL or pAmpC-encoding genes (number of isolates): blaSHV-12 (9), blaCTX-M-1 (3), blaCTX-M-14 (2), and blaCMY-2 (1). The last CTXR isolate presented a −42-point-mutation in the chromosomal ampC promoter. Eleven out of 15 ESBL/pAmpC E. coli isolates were multiresistant (most common resistance phenotype: β-lactams-quinolones-tetracycline-sulfamethoxazole/trimethoprim). A plasmid-mediated quinolone resistance determinant (qnrS1) was identified in one E. coli from a barn owl. High genetic diversity was observed among ESBL/pAmpC E. coli isolates, with 12 different sequence types (STs), including several strains of STs frequently detected among human clinical isolates (ST38/D, ST131/B2, ST155/B1, ST10/A). The ST131 isolate belonged to the emergent ciprofloxacin-resistant H30R subclone. This study reveals a high percentage of bird as carriers of ESBL/pAmpC E. coli isolates in Spain, highlighting the elevated rate among storks, kites, and vultures. Wild birds can contribute to the global spread of ESBL/pAmpC-producing E. coli in natural ecosystems.

First detection of methicillin-resistant Staphylococcus aureus ST398 and Staphylococcus pseudintermedius ST68 from hospitalized equines in Spain.

Eight coagulase‐positive staphylococci from equines with different pathologies obtained between 2005 and 2011 were investigated. Isolates were characterized by different molecular techniques (spa‐, agr‐, MLST), and clonal relatedness of strains was investigated by ApaI and SmaI PFGE. Anti‐microbial resistance and virulence profiles were determined. Six isolates were identified as Staphylococcus aureus, and two as Staphylococcus pseudintermedius. Of these, four isolates were methicillin‐resistant S. aureus (MRSA) ST398 and one S. pseudintermedius was mecA positive and typed as ST68. One MRSA ST398 strain was isolated in 2005 and might be one of the earliest MRSA ST398 descriptions in Spain. All 5 mecA‐positive strains were multidrug resistant and were isolated from hospitalized equines. Three MRSA ST398 strains carried the recently described transposon Tn559 within the chromosomal radC gene. The mecA‐positive S. pseudintermedius ST68 strain was also multidrug resistant and harboured the erm(B)‐Tn5405‐like element. This ST68 strain presented a clear susceptible phenotype to oxacillin and cefoxitin regardless of the presence of an integral and conserved mecA gene and mecA promoter, which enhances the need for testing the presence of this gene in routine analysis to avoid treatment failures. These data reflect the extended anti‐microbial resistance gene acquisition capacities of both bacterial species and evidence their pathogenic properties. The first detection of MRSA ST398 and S. pseudintermedius ST68 in horses in Spain is reported.

The Impact of Climate Trends on a Tick Affecting Public Health: A Retrospective Modeling Approach for Hyalomma marginatum (Ixodidae).

The impact of climate trends during the period 1901–2009 on the life cycle of Hyalomma marginatum in Europe was modeled to assess changes in the physiological processes of this threat to public health. Monthly records of temperature and water vapour at a resolution of 0.5° and equations describing the life cycle processes of the tick were used. The climate in the target region affected the rates of the life cycle processes of H. marginatum: development rates increased, mortality rates in molting stages decreased, and the survival rates of questing ticks decreased in wide territories of the Mediterranean basin. The modeling framework indicated the existence of critical areas in the Balkans, central Europe, and the western coast of France, where the physiological processes of the tick improved to extents that are consistent with the persistence of populations if introduced. A spatially explicit risk assessment was performed to detect candidate areas where active surveys should be performed to monitor changes in tick density or persistence after a hypothetical introduction. We detected areas where the critical abiotic (climate) and biotic (host density) factors overlap, including most of the Iberian peninsula, the Mediterranean coast of France, eastern Turkey, and portions of the western Black Sea region. Wild ungulate densities are unavailable for large regions of the territory, a factor that might affect the outcome of the study. The risk of successfully establishing H. marginatum populations at northern latitudes of its current colonization range seems to be still low, even if the climate has improved the performance of the tick in these areas.

New Approaches for Education and Training in Veterinary Public Health: The SAPUVET Projects

Continued contact between humans and animals, in combination with the ever-increasing movement of human and animal populations that is one effect of globalization, contributes to the spread of diseases, often with detrimental effects on public health. This has led professionals involved in both animal health and public health to recognize veterinary public health (VPH) as a key area for their activities to address the human-animal interface. Veterinarians, a profession with major involvement in this field, are in need of specific knowledge and skills to prevent and control public-health problems. As a result, VPH must be directly integrated into veterinary educational programs. At present, only few veterinary schools have specific VPH programs; in most institutions, VPH does not feature as a specific subject in either undergraduate or post-graduate curricula. SAPUVET and SAPUVETNET II are network projects supported by the ALFA program of the European Union (EU). Their main objectives are to reach a common understanding between European and Latin American universities in the definition of the areas in which VPH is important in their respective countries, and to design a harmonized training program for veterinarians in VPH, by making use of new technological applications and innovative teaching methodologies. The elaboration of educational material in combination with case studies presenting real-life problems provides a basis to apply the knowledge acquired on VPH. It is envisaged that the material and modules developed during the two projects will be integrated into the veterinary curricula of the participating universities, as well as in other partner organizations.