Publications by authors named "Ursula Höfle"

63 Publications

Genomic Analysis of of the Lineage CC130, Including -Carrying MRSA and MSSA Isolates Recovered of Animal, Human, and Environmental Origins.

Front Microbiol 2021 25;12:655994. Epub 2021 Mar 25.

Area of Biochemistry and Molecular Biology, OneHealth-UR Research Group, University of La Rioja, Logroño, Spain.

Most methicillin resistant (MRSA) isolates harboring gene belong to clonal complex CC130. This lineage has traditionally been regarded as animal-associated as it lacks the human specific immune evasion cluster (IEC), and has been recovered from a broad range of animal hosts. Nevertheless, sporadic -MRSA human infections have been reported, with evidence of zoonotic transmission in some cases. The objective of this study was to investigate the whole-genome sequences of 18 CC130 isolates [13 methicillin-resistant (-MRSA) and five methicillin-susceptible (MSSA)] from different sequences types, obtained from a variety of host species and origins (human, livestock, wild birds and mammals, and water), and from different geographic locations, in order to identify characteristic markers and genomic features. Antibiotic resistance genes found among MRSA-CC130 were those associated with the SSCXI element. Most MRSA-CC130 strains carried a similar virulence gene profile. Additionally, six MRSA-CC130 possessed and one MSSA-ST130 had . The MSSA-ST700 strains were most divergent in their resistance and virulence genes. The pan-genome analysis showed that 29 genes were present solely in MRSA-CC130 (associated with SCCXI) and 21 among MSSA-CC130 isolates (associated with phages). The SCCXI, PBP3, GdpP, and AcrB were identical at the amino acid level in all strains, but some differences were found in PBP1, PBP2, PBP4, and YjbH proteins. An examination of the host markers showed that the 3' region of the bacteriophage φ3 was nearly identical to the reference sequence. Truncated gene was also found in -negative strains (two of them carrying -type gene). The gene of wild rabbit isolates included novel mutations. The gene was found in the three MSSA-ST700 strains from small ruminants and in one MSSA-ST130 from a red deer; these strains also carried a -type gene, different from the human and equine variants. Finally, a phylogenetic analysis showed that the three MSSA-ST700 strains and the two MSSA-ST130 strains cluster separately from the remaining MRSA-CC130 strains with the gene as marker for the main lineage. The presence of the human IEC cluster in some -MRSA-CC130 strains suggests that these isolates may have had a human origin.
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http://dx.doi.org/10.3389/fmicb.2021.655994DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8027229PMC
March 2021

A multidisciplinary approach to the evaluation of the effects of foraging on landfills on white stork nestlings.

Sci Total Environ 2021 Feb 10;775:145197. Epub 2021 Feb 10.

SaBio Research Group, Instituto de Investigación en Recursos Cinegéticos IREC, (CSIC-UCLM-JCCM), Ciudad Real, Spain.

The use of landfills as foraging areas by white storks (Ciconia ciconia) is a recent well-known behaviour. While several studies have highlighted positive effects at a populational level others suggest that the presence of pollutants, pathogens and the lower presence of antioxidants in the food could pose a health risk for individuals. The objective of this study was to evaluate potential effects of the use of landfills as a food resource on the physiology and health of white stork nestlings, by a multidisciplinary approach based on the analysis of nutritional status, body condition, blood parameters, oxidative stress balance and the presence of pathogens. Results showed better body condition in individuals associated with landfills compared to the ones feeding on natural resources, as well as better nutritional status, as indicated by higher levels of albumin, cholesterol, and triglycerides in plasma. As many pollutants have a pro-oxidant effect, we evaluated oxidative stress balance, with no differences in the indicators of damage except for methaemoglobin (metHb), significantly higher in nestlings associated with landfill-origin food. Regarding antioxidants, GSH was higher in nestlings associated with landfills, which may suggest a hormetic response induced potentially by the presence of pollutants in waste. Nestlings fed food from landfills also had a higher presence of Escherichia coli with a multiresistant phenotype to antibiotics. In conclusion, our results show that nestlings fed with a higher proportion of food from landfills present a better nutritional status and body condition than those fed with a higher proportion of natural diet, being the only indicators of negative effects of the use of this food resource the higher percentage of metHb in the peripheral blood and the presence of antibiotic-resistant E. coli.
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http://dx.doi.org/10.1016/j.scitotenv.2021.145197DOI Listing
February 2021

Frequency and Characterization of Antimicrobial Resistance and Virulence Genes of Coagulase-Negative Staphylococci from Wild Birds in Spain. Detection of -Carrying Isolates.

Microorganisms 2020 Aug 29;8(9). Epub 2020 Aug 29.

Área de Bioquímica y Biología Molecular, Universidad de La Rioja, 26006 Logroño, Spain.

The objective of this study was to determine the prevalence and diversity of coagulase-negative staphylococci (CoNS) species from wild birds in Spain, as well as to analyze the antimicrobial resistance phenotype/genotype and the virulence gene content. During 2015-2016, tracheal samples of 242 wild birds were collected in different regions of Spain for staphylococci recovery. The species identification was performed using MALDI-TOF. The antimicrobial resistance phenotype and genotype was investigated by the disk diffusion method and by PCR, respectively. The presence of the virulence genes /-PV, , , , and was investigated by PCR. Moreover, CoNS carrying the gene were subjected to SCC typing. Of the tested animals, 60% were CoNS-carriers, and 173 CoNS isolates were recovered from the 146 positive animals, which belonged to 11 species, with predominance of ( = 118) and ( = 25). A total of 34% of CoNS isolates showed a multidrug resistance phenotype, and 42 -positive methicillin-resistant CoNS (MRCoNS) were detected. The isolates showed resistance to the following antimicrobials (percentage of resistant isolates/antimicrobial resistance genes detected): penicillin (49/ , ), cefoxitin (24/ ), erythromycin and/or clindamycin (92/ (B), (C), (43), (A), (C), (A), (B), (A) and (A)), gentamicin and/or tobramycin (5/ (6')-Ie-(2″)-Ia, (4')-Ia), streptomycin (12/), tetracycline (17/ (K), (L), (M)), ciprofloxacin (4), chloramphenicol (1/ ), fusidic acid (86/ , ) and trimethoprim-sulfamethoxazole (1/ ). None of the isolates harbored the /-PV, , , and genes, but two isolates (1%) carried the gene. Wild birds are frequently colonized by CoNS species, especially . We identified scavenging on intensively produced livestock and feeding on landfills as risk factors for CoNS carriage. High proportions of MRCoNS and multidrug resistant CoNS were detected, which coupled with the presence of important virulence genes is of concern.
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http://dx.doi.org/10.3390/microorganisms8091317DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7564563PMC
August 2020

Foraging at Solid Urban Waste Disposal Sites as Risk Factor for Cephalosporin and Colistin Resistant Carriage in White Storks ().

Front Microbiol 2020 28;11:1397. Epub 2020 Jul 28.

IRTA, Centre de Recerca en Sanitat Animal (CReSA, IRTA-UAB), Universitat Autònoma de Barcelona (UAB), Bellaterra, Spain.

White stork () may act as a reservoir and vehicle of cephalosporin resistant (CR) . Between 2011 and 2014, we sampled white storks from colonies exposed to different degrees of anthropic pressure across the major areas of natural distribution of white storks in Spain. Cloacal swab samples ( = 467) were obtained from individuals belonging to 12 different colonies from six different regions. Additionally, 70 samples were collected from recently deposited droppings at the base of nesting platforms. We phenotypically characterized isolates, confirmed presence of CR genes and classified plasmids. Risk factors for acquiring these genes were assessed. Overall, 8.8% (41 out of 467) storks carried CR in their cloaca and five (7.1%) were identified from recently deposited droppings; therefore, 46 isolates were further characterized. Of them, 20 contained , nine , six , four , three , two , one together with , and one together with . All were multidrug-resistant, and four harbored the plasmid-mediated colistin resistance gene. CR genes were associated with the presence of IncI1, IncFIB, and IncN replicon families. I-macrorestriction analysis revealed a great diversity among most of the I-PFGE types, but indistinguishable types were also seen with isolates obtained from different locations. Clonal complex 10 was the most common among CR and two positive isolates were identified as B2-ST131. Carriage of CR was significantly higher in colonies located close to solid urban waste disposal sites in which foraging on human waste was more likely and in one case to cattle grazing. The co-occurrence of and -1 on plasmids of isolated from wild birds as early as 2011 is of note, as the earliest previous report of -1 in wild birds is from 2016. Our study shows that foraging at landfills and in association with cattle grazing are important risk factors for the acquisition of CR in white storks.
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http://dx.doi.org/10.3389/fmicb.2020.01397DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7399022PMC
July 2020

-Colistin Resistance Genes Mobilized by IncX4, IncHI2, and IncI2 Plasmids in of Pigs and White Stork in Spain.

Front Microbiol 2019 17;10:3072. Epub 2020 Jan 17.

Food and Drug Administration, Silver Spring, MD, United States.

Colistin has become the last-line antimicrobial for the treatment of multidrug resistant (MDR) in human medicine. To date, several colistin resistance genes have been described. Of them -1 is disseminated worldwide in of human and animal origin. The aim of this study was to characterize -mediated resistance plasmids from of animal origin in Spain. From our strain collection, 70 of pig origin collected between 2005 and 2014 (10 per year, except for years 2009-2010-2013) were randomly selected and screened for the presence of -genes. Additionally, 20 isolated in 2011 from white storks () from the same urban household waste landfill associated colony were also included. Whole genome sequencing of -positive isolates was carried out on a MiSeq (Illumina). Hybrid whole genome sequencing strategy combining nanopore and Illumina technologies were performed in a selection of isolates to close the genomes and plasmids and identify the presence of antimicrobial resistance genes. Minimum inhibitory concentration (MIC) was used to assess the susceptibility to colistin. Mating experiments were carried out to evaluate transferability of the -genes. A total of 19 -1 and one -4 positive isolates were detected, 15 from pigs distributed during the study period, and five from storks collected in 2011. No other -variants were found. The MICs for colistin ranged between 4 and >4 mg/L. High diversity of STs were detected among the positive isolates, with only ST-10 shared between pigs and white storks. Except for one isolate, all were genotypic and phenotypically MDR, and five of them also harbored cephalosporin resistance genes (, , and three ). -1 genes were mobilizable by conjugation, associated with IncX4, IncHI2, and IncI2 plasmids. In our study, -1 genes have been circulating in pig farms since 2005 harbored by a variety of clones. Its persistence may be driven by co-selection since plasmids containing -1 also exhibit resistance to multiple drugs used in veterinary medicine. Furthermore, this is the first report of the presence of -1 gene in isolates from white storks in Spain. This finding highlights the potential importance of wildlife that forage at urban household waste landfills in the transmission and spread of colistin resistance genes.
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http://dx.doi.org/10.3389/fmicb.2019.03072DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978640PMC
January 2020

Antioxidant supplementation slows telomere shortening in free-living white stork chicks.

Proc Biol Sci 2020 01 15;287(1918):20191917. Epub 2020 Jan 15.

Groningen Institute for Evolutionary Life Sciences, University of Groningen, Groningen, The Netherlands.

Telomere length (TL) and shortening is increasingly shown to predict variation in survival and lifespan, raising the question of what causes variation in these traits. Oxidative stress is well known to accelerate telomere attrition , but its importance is largely hypothetical. We tested this hypothesis experimentally by supplementing white stork () chicks with antioxidants. Individuals received either a control treatment, or a supply of tocopherol (vitamin E) and selenium, which both have antioxidant properties. The antioxidant treatment increased the concentration of tocopherol for up to two weeks after treatment but did not affect growth. Using the telomere restriction fragment technique, we evaluated erythrocyte TL and its dynamics. Telomeres shortened significantly over the 21 days between the baseline and final sample, independent of sex, mass, size and hatching order. The antioxidant treatment significantly mitigated shortening rate of average TL (-31% in shorter telomeres; percentiles 10th, 20th and 30th). Thus, our results support the hypothesis that oxidative stress shortens telomeres .
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http://dx.doi.org/10.1098/rspb.2019.1917DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7003462PMC
January 2020

A vaccinology Approach to the Identification and Characterization of Candidate Protective Antigens for the Control of Poultry Red Mite Infestations.

Vaccines (Basel) 2019 Nov 20;7(4). Epub 2019 Nov 20.

SaBio. Instituto de Investigación en Recursos Cinegéticos, IREC (CSIC-UCLM-JCCM), Ronda de Toledo 12, 13071, Ciudad Real, Spain.

The poultry red mite (PRM), , is a hematophagous ectoparasite considered as the major pest in the egg-laying industry. Its pesticide-based control is only partially successful and requires the development of new control interventions such as vaccines. In this study, we follow a vaccinology approach to identify PRM candidate protective antigens. Based on proteomic data from fed and unfed nymph and adult mites, we selected a novel PRM protein, calumenin (Deg-CALU), which is tested as a vaccine candidate on an on-hen trial. Subolesin (Rhm-SUB) was chosen as a positive control. Deg-CALU and Rhm-SUB reduced the mite oviposition by 35 and 44%, respectively. These results support Deg-CALU and Rhm-SUB as candidate protective antigens for the PRM control.
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http://dx.doi.org/10.3390/vaccines7040190DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6963798PMC
November 2019

Reduction in Oviposition of Poultry Red Mite () in Hens Vaccinated with Recombinant Akirin.

Vaccines (Basel) 2019 Sep 19;7(3). Epub 2019 Sep 19.

SaBio, Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), Ronda de Toledo 12, 13071 Ciudad Real, Spain.

The poultry red mite (PRM), , is a hematophagous ectoparasite of birds with worldwide distribution that causes economic losses in the egg-production sector of the poultry industry. Traditional control methods, mainly based on acaricides, have been only partially successful, and new vaccine-based interventions are required for the control of PRM. Vaccination with insect Akirin (AKR) and its homolog in ticks, Subolesin (SUB), have shown protective efficacy for the control of ectoparasite infestations and pathogen infection/transmission. The aim of this study was the identification of the akr gene from (), the production of the recombinant Deg-AKR protein, and evaluation of its efficacy as a vaccine candidate for the control of PRM. The anti-Deg-AKR serum IgY antibodies in hen sera and egg yolk were higher in vaccinated than control animals throughout the experiment. The results demonstrated the efficacy of the vaccination with Deg-AKR for the control of PRM by reducing mite oviposition by 42% following feeding on vaccinated hens. A negative correlation between the levels of serum anti-Deg-AKR IgY and mite oviposition was obtained. These results support Deg-AKR as a candidate protective antigen for the control of PRM population growth.
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http://dx.doi.org/10.3390/vaccines7030121DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6789658PMC
September 2019

A Recombinant Subviral Particle-Based Vaccine Protects Magpie () Against West Nile Virus Infection.

Front Microbiol 2019 5;10:1133. Epub 2019 Jun 5.

Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain.

The mosquito-borne West Nile virus (WNV) is a highly neurovirulent currently representing an emergent zoonotic concern. WNV cycles in nature between mosquito vectors and birds that act as amplifier hosts and play an essential role in virus ecology, being, thus, WNV a threat to many species. Availability of an efficient avian vaccine would benefit certain avian populations, both birds grown for hunting and restocking activities, as well as endangered species in captive breeding projects, wildlife reservations, and recreation installations, and would be useful to prevent and contain outbreaks. Avian vaccination would be also of interest to limit WNV spillover to humans or horses from susceptible bird species that live in urbanized landscapes, like magpies. Herein, we have addressed the efficacy of a single dose of a WNV recombinant subviral particle (RSP) vaccine in susceptible magpie (). The protective capacity of the RSP-based vaccine was demonstrated upon challenge of magpies with 5 × 10 plaque forming units of a neurovirulent WNV strain. A significant improvement in survival rates of immunized birds was recorded when compared to vehicle-inoculated animals (71.4 vs. 22.2%, respectively). Viremia, which is directly related to the capacity of a host to be competent for virus transmission, was reduced in vaccinated animals, as was the presence of infectious virus in feather follicles. Bird-to-bird transmission was recorded in three of six unchallenged (contact) magpies housed with non-vaccinated WNV-infected birds, but not in contact animals housed with vaccinated WNV-infected magpies. These results demonstrate the protective efficacy of the RSP-based vaccine in susceptible birds against WNV infection and its value in controlling the spread of the virus.
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http://dx.doi.org/10.3389/fmicb.2019.01133DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6560071PMC
June 2019

Naturally Avian Influenza Virus-Infected Wild Birds Are More Likely to Test Positive for spp. and spp.

Avian Dis 2019 03;63(sp1):131-137

NEIKER-Instituto Vasco de Investigación y Desarrollo Agrario, Animal Health Department, Bizkaia Science and Technology Park 812L, 48160 Derio (Bizkaia), Spain,

Wild birds often harbor infectious microorganisms. Some of these infectious microorganisms may present a risk to domestic animals and humans through spillover events. Detections of certain microorganisms have been shown to increase host susceptibility to infections by other microorganisms, leading to coinfections and altered host-to-host transmission patterns. However, little is known about the frequency of coinfections and its impact on wild bird populations. In order to verify whether avian influenza virus (AIV) natural infection in wild waterbirds was related to the excretion of other microorganisms, 73 AIV-positive samples (feces and cloacal swabs) were coupled with 73 AIV-negative samples of the same sampling characteristics and tested by real-time PCR specific for the following microorganisms: West Nile virus, avian avulavirus 1, spp., , , subspecies, complex, and spp. Concurrent detections were found in 47.9% (35/73) of the AIV-positive samples and in 23.3% (17/73) of the AIV-negative samples ( = 0.003). spp. and spp. were found to be significantly more prevalent among the AIV-positive samples than among the AIV-negative samples (42.9% . 22.8%; = 0.024 and 15.2% . 0.0%; = 0.0015, respectively). Prevalence of concurrent detections differed significantly among sampling years ( = 0.001), host families ( = 0.002), host species ( = 0.003), AIV subtypes ( = 0.003), and type of sample ( = 0.009). Multiple concurrent detections (more than one of the tested microorganisms excluding AIV) were found in 9.6% (7/73) of all the AIV-positive samples, accounting for 20% (7/35) of the concurrent detection cases. In contrast, in AIV-negative samples we never detected more than one of the selected microorganisms. These results show that AIV detection was associated with the detection of the monitored microorganisms. Further studies of a larger field sample set or under experimental conditions are necessary to infer causality in these trends.
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http://dx.doi.org/10.1637/11866-042518-Reg.1DOI Listing
March 2019

Detection of MRSA of Lineages CC130-mecC and CC398-mecA and Staphylococcus delphini-lnu(A) in Magpies and Cinereous Vultures in Spain.

Microb Ecol 2019 Aug 29;78(2):409-415. Epub 2019 Jan 29.

Área de Bioquímica y Biología Molecular, Universidad de La Rioja, Madre de Dios 51, 26006, Logroño, Spain.

The aim of this study was to determine the carriage rate of coagulase-positive staphylococci (CoPS) in wild birds and to characterize recovered isolates. Tracheal samples from 324 wild birds, obtained in different Spanish regions during 2015-2016, were screened for CoPS carriage. The antimicrobial resistance profile and the virulence gene content were investigated. Molecular typing was performed by spa, agr, MLST, SCCmec, and S. delphini group classification. CoPS were recovered from 26 samples of wild birds (8.3%), and 27 isolates were further characterized. Two CoPS species were detected: S. aureus (n = 15; eight cinereous vultures and seven magpies) and S. delphini (n = 12; 11 cinereous vultures and one red kite). Thirteen S. aureus were methicillin-resistant (MRSA) and the remaining two strains were methicillin-susceptible (MSSA). Twelve MRSA were mecC-positive, typed as t843-ST1583/ST1945/ST1581/ST1571 (n = 11) and t1535-ST1945 (n = 1) (all of clonal-complex CC130); they were susceptible to the non-β-lactams tested. The remaining MRSA strain carried the mecA gene, was typed as t011-ST398-CC398-agrI-SCCmec-V, and showed a multiresistance phenotype. MSSA isolates were ascribed to lineages ST97-CC97 and ST425-CC425. All S. aureus lacked the studied virulence genes (lukS/F-PV, tst, eta, etb, and etd), and the IEC type E (with scn and sak genes) was detected in four mecC-positive and one MSSA isolates. S. delphini strains were methicillin-susceptible but showed resistance to at least one of the antimicrobials tested, with high penicillin (75%, with blaZ gene) and tetracycline [58%, with tet(K)± tet(L)] resistance rates. All S. delphini isolates presented the virulence genes lukS-I, siet, and se-int, and four carried the clindamycin-resistance lnu(A) gene.
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http://dx.doi.org/10.1007/s00248-019-01328-4DOI Listing
August 2019

Long-term avian influenza virus epidemiology in a small Spanish wetland ecosystem is driven by the breeding Anseriformes community.

Vet Res 2019 Jan 17;50(1). Epub 2019 Jan 17.

Animal Health Department, NEIKER-Instituto Vasco de Investigación y Desarrollo Agrario, Parque tecnológico de Bizkaia P-812, 48160, Derio, Bizkaia, Spain.

During 2007-2009 and 2012-2014, avian influenza virus (AIV) was studied in a wild avian community of a northern Spanish wetland using non-invasive sampling methods and host identification by COI barcoding. The aim of this longitudinal study was to evaluate AIV dynamics in a natural wetland ecosystem, taking into account both virological aspects and ecological traits of hosts. Global AIV prevalence decreased significantly during the second sampling period (0.3%) compared to the first (6.6%). Circulating subtype distributions were also different between periods, with a noteworthy H5 and H7 subtype richness during the first sampling period. Mallard Anas platyrhynchos was identified as the main AIV host, although not all positive samples could be ascribed to the host. We modelled AIV prevalence with regard to the avian host community composition and meteorological data from the wetland. Statistical analysis revealed seasonal differences in AIV detection, with higher prevalence during the breeding season compared to other phenological events. The model also shows that the lower AIV prevalence during the second study period was associated with a significant reduction of breeding Anseriformes in the wetland, revealing a long-term fluctuation of AIV prevalence driven by the breeding Anseriformes community. This longitudinal study on AIV epidemiology in a natural ecosystem reveals that although prevalence follows seasonal and annual patterns, long-term prevalence fluctuation is linked to the breeding community composition and size. These results are relevant to understanding the influence of host ecology on pathogen transmission for preventing and managing influenza emergence.
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http://dx.doi.org/10.1186/s13567-019-0623-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6337815PMC
January 2019

High susceptibility of magpie (Pica pica) to experimental infection with lineage 1 and 2 West Nile virus.

PLoS Negl Trop Dis 2018 04 10;12(4):e0006394. Epub 2018 Apr 10.

Departamento de Biotecnología. INIA, Madrid, Spain.

West Nile virus (WNV), a zoonotic pathogen naturally transmitted by mosquitoes whose natural hosts are birds, has spread worldwide during the last few decades. Resident birds play an important role in flavivirus epidemiology, since they can serve as reservoirs and facilitate overwintering of the virus. Herein, we report the first experimental infection of magpie (Pica pica) with two strains of West Nile virus, lineages 1 (NY-99) and 2 (SRB Novi-Sad/12), which are currently circulating in Europe. Magpies were highly susceptible to WNV infection, with similar low survival rates (30% and 42.8%) for both lineages. All infected magpies developed viremia detectable at 3 days post-infection with titers above those necessary for successful transmission of WNV to a mosquito. Neutralizing antibodies were detected at all time points analyzed (from 7 to 17 days post-infection). WNV genome was detected in the brains and hearts of all magpies that succumbed to the infection, and, in some of the surviving birds. WNV-RNA was amplified from swabs (oral and cloacal) at 3, 6 and 7 days post-infection and feather pulps, from 3 to 17 days post-infection, of infected animals. Even more, infectious virus was recovered from swabs up to 7 days post-infection and from feather pulps up to 10 days post infection. Sham-infected control animals were negative for viremia, viral RNA, and antibodies. These results suggest that the magpie, which is one of the most abundant corvid species in Europe, could represent a source of WNV transmission for birds and humans. Our observations shed light on the pathogenesis, transmission, and ecology of WNV and can benefit the implementation of surveillance and control programs.
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http://dx.doi.org/10.1371/journal.pntd.0006394DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5909923PMC
April 2018

Absence of protection from West Nile virus disease and adverse effects in red legged partridges after non-structural NS1 protein administration.

Comp Immunol Microbiol Infect Dis 2018 Feb 9;56:30-33. Epub 2018 Jan 9.

Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria - Centro de Investigación en Sanidad Animal (INIA-CISA), Ctra Algete-El Casar s/n, 208130, Valdeolmos, Madrid, Spain; Centro de Investigación Biomédica en Red de Epidemiologia y Salud Pública (CIBERESP), Spain. Electronic address:

The red-legged partridge (Alectoris rufa) is a competent host for West Nile virus (WNV) replication and highly susceptible to WNV disease. With the aim to assess in this species whether the inoculation of non-structural protein NS1 from WNV elicits a protective immune response against WNV infection, groups of partridges were inoculated with recombinant NS1 (NS1 group) or an unrelated recombinant protein (mock group), and challenged with infectious WNV. A third group received no inoculation prior to challenge (challenge group). The NS1 group failed to elicit detectable antibodies to NS1 while in the mock group a specific antibody response was observed. Moreover, no protection against WNV disease was observed in the NS1 group, but rather, it showed significantly higher viral RNA load and delayed neutralizing antibody response, and suffered a more severe clinical disease, which resulted in higher mortality. This adverse effect has not been observed before and warrants further investigations.
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http://dx.doi.org/10.1016/j.cimid.2017.12.006DOI Listing
February 2018

Avian Influenza Virus Surveillance in South-Central Spain Using Fecal Samples of Aquatic Birds Foraging at Landfills.

Front Vet Sci 2017 23;4:178. Epub 2017 Oct 23.

SaBio Working Group, Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), Ciudad Real, Spain.

Aquatic wild birds have been intensively studied to better understand their role in avian influenza virus (AIV) maintenance and spread. To date, AIV surveillance has primarily focused on natural aquatic environments where different bird species aggregate and viral survival is enhanced. However, artificial habitats such as landfills are attracting substantial numbers of wild birds, AIV reservoir species included. The use of landfills as a predictable food source has significantly influenced population size, migratory traits, and feeding behavior of white storks () and black-headed gulls () among others. Considering the proximity of landfills to urban settlements and frequently poultry-farms, targeted monitoring of AIV in bird species that forage at landfills but are known to also frequent urban and agricultural habitats could be a useful means for monitoring of AIV, especially during periods of bird aggregation. During the wintering season 2014-2015, the prevalence of AIV in five avian species at two landfills in South-Central Spain was explored by rRT-PCR and species related temporal variation in AIV prevalence determined. We collected and tested 1,186 fresh fecal samples from white storks ( = 689), cattle egrets ( = 116) and mixed flocks of gulls ( = 381) as well as cloacal and oral swabs from five birds found dead. Seven samples contained AIV, five from gulls and one each from a stork and a cattle egret. Overall, AIV prevalence was 0.60%. No significant temporal variation was observed in AIV prevalence. Prevalence differed significantly among the sampled taxonomic groups, being highest in gulls (1.31%). H16N3 subtype was detected from a cattle egret and H11N9 subtype from a white stork, whereas gulls harbored both subtypes in addition to H11N3 subtype. H16 subtype detection in a cattle egret evidences its host range may not be restricted to gulls. Our results indicate that wild birds foraging at landfills may carry different LPAIV subtypes.
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http://dx.doi.org/10.3389/fvets.2017.00178DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5662893PMC
October 2017

Spatio-temporal trends and risk factors affecting West Nile virus and related flavivirus exposure in Spanish wild ruminants.

BMC Vet Res 2016 Nov 9;12(1):249. Epub 2016 Nov 9.

Instituto de Investigación en Recursos Cinegéticos IREC, (CSIC-UCLM-JCCM), Ciudad Real, Spain.

Background: During the last decade, the spread of many flaviviruses (Genus Flavivirus) has been reported, representing an emerging threat for both animal and human health. To further study utility of wild ruminant samples in West Nile virus (WNV) surveillance, we assessed spatio-temporal trends and factors associated with WNV and cross-reacting flaviviruses exposure, particularly Usutu virus (USUV) and Meaban virus (MBV), in wild ruminants in Spain. Serum samples from 4693 wild ruminants, including 3073 free-living red deer (Cervus elaphus), 201 fallow deer (Dama dama), 125 mouflon (Ovis aries musimon), 32 roe deer (Capreolus capreolus) and 1262 farmed red deer collected in 2003-2014, were screened for WNV and antigenically-related flavivirus antibodies using a blocking ELISA (bELISA). Positive samples were tested for neutralizing antibodies against WNV, USUV and MBV by virus micro-neutralization tests.

Results: Mean flavivirus seroprevalence according to bELISA was 3.4 ± 0.5 % in red deer, 1.0 ± 1.4 % in fallow deer, 2.4 ± 2.7 % in mouflon and 0 % in roe deer. A multivariate logistic regression model revealed as main risk factors for seropositivity in red deer; year (2011), the specific south-coastal bioregion (bioregion 5) and presence of wetlands. Red deer had neutralizing antibodies against WNV, USUV and MBV.

Conclusions: The results indicate endemic circulation of WNV, USUV and MBV in Spanish red deer, even in areas without known flavivirus outbreaks. WNV antibodies detected in a free-living red deer yearling sampled in 2010, confirmed circulation this year. Co-circulation of WNV and USUV was detected in bioregions 3 and 5, and of WNV and MBV in bioregion 3. Sampling of hunted and farmed wild ruminants, specifically of red deer yearlings, could be a complementary way to national surveillance programs to monitor the activity of emerging flaviviruses.
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http://dx.doi.org/10.1186/s12917-016-0876-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5103426PMC
November 2016

Characterization of fecal vancomycin-resistant enterococci with acquired and intrinsic resistance mechanisms in wild animals, Spain.

Microb Ecol 2016 11 21;72(4):813-820. Epub 2015 Jul 21.

Área Bioquímica y Biología Molecular, Universidad de La Rioja, Madre de Dios 51, 26006, Logroño, Spain.

The objectives were to evaluate the presence of vancomycin-resistant enterococci with acquired (VRE-a) and intrinsic (VRE-i) resistance mechanisms in fecal samples from different wild animals, and analyze their phenotypes and genotypes of antimicrobial resistance. A total of 348 cloacal/rectal samples from red-legged partridges (127), white storks (81), red kites (59), and wild boars (81) (June 2014/February 2015) were inoculated in Slanetz-Bartley agar supplemented with vancomycin (4 μg/mL). We investigated the susceptibility to 12 antimicrobials and the presence of 19 antimicrobial resistance and five virulence genes. In addition, we performed multilocus sequence typing, detection of IS16 and studied Tn1546 structure. One VRE-a isolate was identified in one wild boar. This isolate was identified as Enterococcus faecium, harbored vanA gene included into Tn1546 (truncated with IS1542/IS1216), and belonged to the new ST993. This isolate contained the erm(A), erm(B), tet(M), dfrG, and dfrK genes. Neither element IS16 nor the studied virulence genes were detected. Ninety-six VRE-i isolates were identified (89 Enterococcus gallinarum and seven Enterococcus casseliflavus), with the following prevalence: red kites (71.2 %), white storks (46.9 %), red-legged partridges (7.9 %), and wild boars (4.9 %). Most E. gallinarum isolates showed resistance to tetracycline (66.3 %) and/or erythromycin (46.1 %). High-level resistance to aminoglycosides was present among our VRE-i isolates: kanamycin (22.9 %), streptomycin (11.5 %), and gentamicin (9.4 %). In general, VRE-i isolates of red kites showed higher rates of resistance for non-glycopeptide agents than those of other animal species. The dissemination of acquired resistance mechanisms in natural environments could have implications in the global spread of resistance with public health implications.
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http://dx.doi.org/10.1007/s00248-015-0648-xDOI Listing
November 2016

Use of wildlife rehabilitation centres in pathogen surveillance: A case study in white storks (Ciconia ciconia).

Prev Vet Med 2016 Aug 21;130:106-11. Epub 2016 Jun 21.

SaBio Working Group, Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), Ronda de Toledo 12, 13005 Ciudad Real, Spain. Electronic address:

More than 70% of new human pathogens are zoonotic and many originate from the wildlife reservoir. Wildlife rehabilitation centres (WRC) are an easily accessible source for sample and data collection for preventive surveillance, but data collected this way may be biased. We use white storks (Ciconia ciconia) as a model to compare pathogen prevalence obtained in the field and WRC. We address factors that may affect disease prevalence data like origin, the age group and the "diseased" state of WRC admissions. In this study we compared prevalence of Escherichia coli and Salmonella spp. in the digestive tract; antibodies against West Nile virus, avian influenza and Newcastle disease virus, and antimicrobial resistance patterns of E. coli between nestling and adult wild storks established in different habitats (n=90) and storks admitted to two different WRC (n=30) in the same region. When age groups and colonies of origin were disregarded, the mean enterobacteria (E. coli, Salmonella) and viral antibody prevalence of the wild population (n=90) were similar to prevalence observed in the individuals admitted to WRC (n=30). However, in fledgling juvenile storks admitted to WRC, the prevalence of Salmonella spp. (13.3%), E. coli showing resistance to cefotaxime (37.9%) and against two antimicrobials at once (41.4%) were more similar to the prevalence in stork nestlings from landfill-associated colonies (7.9%, 37.1% and 48.6%, respectively for prevalence of Salmonella spp. and E. coli displaying, cefotaxime resistance and resistance against two antimicrobials), and significantly higher than in colonies located in natural habitats (0%; 10.5% and 15.8%, respectively). Thus, pathogen surveillance in individuals from an abundant species admitted to WRC is useful to monitor overall mean prevalence, but for certain pathogens may not be sufficient to detect differences between local populations. In addition, the ecology of the tested species and the specific temporal, spatial and age group distribution of WRC admissions have to be taken into account.
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http://dx.doi.org/10.1016/j.prevetmed.2016.06.012DOI Listing
August 2016

Detection of MRSA ST3061-t843-mecC and ST398-t011-mecA in white stork nestlings exposed to human residues.

J Antimicrob Chemother 2016 Jan 21;71(1):53-7. Epub 2015 Oct 21.

Area of Biochemistry and Molecular Biology, University of La Rioja, Logroño, Spain

Objectives: The objective of this study was to analyse the prevalence of tracheal carriage of Staphylococcus aureus/MRSA in storks and to study the resistance and virulence genes in the obtained isolates.

Methods: Tracheal samples from 92 stork nestlings of two landfill-associated and two natural-habitat colonies were inoculated in specific media for S. aureus and MRSA recovery. Antimicrobial susceptibility was tested, and the presence of resistance, virulence and immune evasion cluster (IEC) genes was analysed by PCR. S. aureus isolates were characterized by spa and agr typing. Staphylococcal cassette chromosome (SCC) mec type was determined for mecC-positive isolates, and MLST was performed for 17 selected S. aureus isolates.

Results: S. aureus isolates were identified in 32/92 samples (34.8%), and 38 isolates were recovered. The prevalence of S. aureus was higher in nestlings from landfills (24/43, 55.8%) than in those from natural habitats (8/49, 16.3%). Three birds from landfill-associated colonies carried MRSA, two with mecA-positive strains [clonal complex (CC) 5-spa-t002 and CC398-spa-t011] and one with a mecC-positive strain [sequence type (ST) 3061-CC130-spa-t843-agr-III-SCCmecXI). None of the MRSA isolates presented IEC genes. Thirty-five MSSA isolates, which showed 18 different spa types (ascribed to CC5, CC7, CC22, CC30, CC45, CC59, CC133 and CC398), were obtained. The agr types detected were I (63%), II (29%) and III (8%). Resistance and virulence genes identified in MSSA were blaZ (n = 25), erm(T) (n = 9), erm(A) (n = 1), tet(M) (n = 2), fexA (n = 3), str (n = 2), tst (n = 2), eta (n = 1) and cna (n = 15). The IEC types B, C, D and G were found in MSSA isolates, and two new STs were identified (ST3060 and ST3061).

Conclusions: White storks are frequently tracheal carriers of S. aureus, including ST398 isolates. MRSA isolates of lineages CC398-mecA and CC130-mecC were detected in storks from landfill-associated colonies exposed to human residues.
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http://dx.doi.org/10.1093/jac/dkv314DOI Listing
January 2016

Long-Term Effect of Serial Infections with H13 and H16 Low-Pathogenic Avian Influenza Viruses in Black-Headed Gulls.

J Virol 2015 Nov 2;89(22):11507-22. Epub 2015 Sep 2.

Department of Viroscience, Erasmus MC, Rotterdam, the Netherlands

Unlabelled: Infections of domestic and wild birds with low-pathogenic avian influenza viruses (LPAIVs) have been associated with protective immunity to subsequent infection. However, the degree and duration of immunity in wild birds from previous LPAIV infection, by the same or a different subtype, are poorly understood. Therefore, we inoculated H13N2 (A/black-headed gull/Netherlands/7/2009) and H16N3 (A/black-headed gull/Netherlands/26/2009) LPAIVs into black-headed gulls (Chroicocephalus ridibundus), their natural host species, and measured the long-term immune response and protection against one or two reinfections over a period of >1 year. This is the typical interval between LPAIV epizootics in wild birds. Reinfection with the same virus resulted in progressively less virus excretion, with complete abrogation of virus excretion after two infections for H13 but not H16. However, reinfection with the other virus affected neither the level nor duration of virus excretion. Virus excretion by immunologically naive birds did not differ in total levels of excreted H13 or H16 virus between first- and second-year birds, but the duration of H13 excretion was shorter for second-year birds. Furthermore, serum antibody levels did not correlate with protection against LPAIV infection. LPAIV-infected gulls showed no clinical signs of disease. These results imply that the epidemiological cycles of H13 and H16 in black-headed gulls are relatively independent from each other and depend mainly on infection of first-year birds.

Importance: Low-pathogenic avian influenza viruses (LPAIVs) circulate mainly in wild water birds but are occasionally transmitted to other species, including humans, where they cause subclinical to fatal disease. To date, the effect of LPAIV-specific immunity on the epidemiology of LPAIV in wild birds is poorly understood. In this study, we investigated the effect of H13 and H16 LPAIV infection in black-headed gulls on susceptibility and virus excretion of subsequent infection with the same or the other virus within the same breeding season and between breeding seasons. These are the only two LPAIV hemagglutinin subtypes predominating in this species. The findings suggest that H13 and H16 LPAIV cycles in black-headed gull populations are independent of each other, indicate the importance of first-year birds in LPAIV epidemiology, and emphasize the need for alternatives to avian influenza virus (AIV)-specific serum antibodies as evidence of past LPAIV infection and correlates of protection against LPAIV infection in wild birds.
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http://dx.doi.org/10.1128/JVI.01765-15DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4645650PMC
November 2015

Identification and characterization of a novel tick-borne flavivirus subtype in goats (Capra hircus) in Spain.

J Gen Virol 2015 Jul 20;96(Pt 7):1676-81. Epub 2015 Feb 20.

1Animal and Plant Health Agency (APHA), Woodham Lane, New Haw, Surrey, KT15 3NB, UK 6Department of Clinical Infection, Microbiology and Immunology, University of Liverpool, Liverpool L69 7BE, UK 7NIHR Health Protection Research Unit in Emerging and Zoonotic Infections, University of Liverpool, Liverpool L69 7BE, UK.

In 2011, a neurological disease was reported in a herd of goats (Capra hircus) in Asturias, Spain. Initial sequencing identified the causative agent as louping ill virus (LIV). Subsequently, with the application of whole genome sequencing and phylogenetic analysis, empirical data demonstrates that the LIV-like virus detected is significantly divergent from LIV and Spanish sheep encephalitis virus (SSEV). This virus encoded an amino acid sequence motif at the site of a previously identified marker for differentiating tick-borne flaviviruses that was shared with a virus previously isolated in Ireland in 1968. The significance of these observations reflects the diversity of tick-borne flaviviruses in Europe. These data also contribute to our knowledge of the evolution of tick-borne flaviviruses and could reflect the movement of viruses throughout Europe. Based on these observations, the proposed name for this virus is Spanish goat encephalitis virus (SGEV), to distinguish it from SSEV.
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http://dx.doi.org/10.1099/vir.0.000096DOI Listing
July 2015

Pathology and tissue tropism of natural West Nile virus infection in birds: a review.

Vet Res 2013 Jun 3;44:39. Epub 2013 Jun 3.

SaBio Instituto de Investigación en Recursos Cinegéticos IREC, (CSIC-UCLM-JCCM) Ronda de Toledo s/n, Ciudad Real 13005, Spain.

West Nile virus (WNV) is a globally distributed arthropod-borne flavivirus capable of infecting a wide variety of vertebrates, with birds as its natural reservoir. Although it had been considered a pathogen of little importance for birds, from the 1990's, and especially after its introduction in the North American continent in 1999, thousands of birds have succumbed to West Nile infection. This review summarizes the pathogenesis and pathology of WNV infection in birds highlighting differences in lesion and antigen distribution and severity among bird orders and families. Despite significant species differences in susceptibility to infection, WNV associated lesions and viral antigen are present in the majority of organs of infected birds. The non-progressive, acute or more prolonged course of the disease accounts for part of the differences in lesion and viral antigen distribution and lesion severity. Most likely a combination of host variables and environmental factors in addition to the intrinsic virulence and pathogenicity of the infecting WNV strain influence the pathogenesis of the infection.
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http://dx.doi.org/10.1186/1297-9716-44-39DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3686667PMC
June 2013

Environmental factors influencing the prevalence of a Clostridium botulinum type C/D mosaic strain in nonpermanent Mediterranean wetlands.

Appl Environ Microbiol 2013 Jul 3;79(14):4264-71. Epub 2013 May 3.

Instituto de Investigación en Recursos Cinegéticos (IREC), CSIC-UCLM-JCCM, Ciudad Real, Spain.

Between 1978 and 2008, 13 avian botulism outbreaks were recorded in the wetlands of Mancha Húmeda (central Spain). These outbreaks caused the deaths of around 20,000 birds from over 50 species, including globally endangered white-headed ducks (Oxyura leucoceophala). Here, a significant association was found between the number of dead birds recorded in each botulism outbreak and the mean temperature in July (always >26°C). The presence of Clostridium botulinum type C/D in wetland sediments was detected by real-time PCR (quantitative PCR [qPCR]) in 5.8% of 207 samples collected between 2005 and 2008. Low concentrations of Cl(-) and high organic matter content in sediments were significantly associated with the presence of C. botulinum. Seventy-five digestive tracts of birds found dead during botulism outbreaks were analyzed; C. botulinum was present in 38.7% of them. The prevalence of C. botulinum was 18.2% (n = 22 pools) in aquatic invertebrates (Chironomidae and Corixidae families) and 33.3% (n = 18 pools) in necrophagous invertebrates (Sarcophagidae and Calliphoridae families), including two pools of adult necrophagous flies collected around bird carcasses. The presence of the bacteria in the adult fly form opens up new perspectives in the epidemiology of avian botulism, since these flies may be transporting C. botulinum from one carcass to another.
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http://dx.doi.org/10.1128/AEM.01191-13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3697518PMC
July 2013

Pathobiology and transmission of highly and low pathogenic avian influenza viruses in European quail (Coturnix c. coturnix).

Vet Res 2013 Mar 28;44:23. Epub 2013 Mar 28.

Centre de Recerca en Sanitat Animal (CReSA), UAB-IRTA, Campus de la Universitat Autònoma de Barcelona, Bellaterra, (Cerdanyola del Vallès), 08193, Spain.

European quail (Coturnix c. coturnix) may share with Japanese quail (Coturnix c. japonica) its potential as an intermediate host and reservoir of avian influenza viruses (AIV). To elucidate this question, European quail were experimentally challenged with two highly pathogenic AIV (HPAIV) (H7N1/HP and H5N1/HP) and one low pathogenic AIV (LPAIV) (H7N2/LP). Contact animals were also used to assess the viral transmission among birds. Severe neurological signs and mortality rates of 67% (H7N1/HP) and 92% (H5N1/HP) were observed. Although histopathological findings were present in both HPAIV-infected groups, H5N1/HP-quail displayed a broader viral antigen distribution and extent of microscopic lesions. Neither clinical nor pathological involvement was observed in LPAIV-infected quail. Consistent long-term viral shedding and effective transmission to naive quail was demonstrated for the three studied AIV. Drinking water arose as a possible transmission route and feathers as a potential origin of HPAIV dissemination. The present study demonstrates that European quail may play a major role in AI epidemiology, highlighting the need to further understand its putative role as an intermediate host for avian/mammalian reassortant viruses.
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http://dx.doi.org/10.1186/1297-9716-44-23DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3640924PMC
March 2013

Worldwide phylogenetic relationship of avian poxviruses.

J Virol 2013 May 13;87(9):4938-51. Epub 2013 Feb 13.

Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary.

Poxvirus infections have been found in 230 species of wild and domestic birds worldwide in both terrestrial and marine environments. This ubiquity raises the question of how infection has been transmitted and globally dispersed. We present a comprehensive global phylogeny of 111 novel poxvirus isolates in addition to all available sequences from GenBank. Phylogenetic analysis of the Avipoxvirus genus has traditionally relied on one gene region (4b core protein). In this study we expanded the analyses to include a second locus (DNA polymerase gene), allowing for a more robust phylogenetic framework, finer genetic resolution within specific groups, and the detection of potential recombination. Our phylogenetic results reveal several major features of avipoxvirus evolution and ecology and propose an updated avipoxvirus taxonomy, including three novel subclades. The characterization of poxviruses from 57 species of birds in this study extends the current knowledge of their host range and provides the first evidence of the phylogenetic effect of genetic recombination of avipoxviruses. The repeated occurrence of avian family or order-specific grouping within certain clades (e.g., starling poxvirus, falcon poxvirus, raptor poxvirus, etc.) indicates a marked role of host adaptation, while the sharing of poxvirus species within prey-predator systems emphasizes the capacity for cross-species infection and limited host adaptation. Our study provides a broad and comprehensive phylogenetic analysis of the Avipoxvirus genus, an ecologically and environmentally important viral group, to formulate a genome sequencing strategy that will clarify avipoxvirus taxonomy.
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http://dx.doi.org/10.1128/JVI.03183-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3624294PMC
May 2013

Tissue tropism and pathology of natural influenza virus infection in black-headed gulls (Chroicocephalus ridibundus).

Avian Pathol 2012 Dec;41(6):547-53

Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), Ciudad Real, 13005, Ciudad Real, Spain.

Black-headed gulls (Chroicocephalus ridibundus) are a suitable host species to study the epidemiology of low-pathogenic avian influenza virus (LPAIV) infection in wild waterbirds because they are a common colony-breeding species in which LPAIV infection is detected frequently, limited mainly to the H13 and H16 subtypes. However, the sites of virus replication and associated lesions are poorly understood. We therefore performed virological and pathological analyses on tissues of black-headed gulls naturally infected with LPAIV. We found that 24 of 111 black-headed gulls collected from breeding colonies were infected with LPAIV (10 birds with H16N3, one bird with H13N8, 13 birds undetermined), based on virus and viral genome detection in pharyngeal and cloacal swabs. Of these 24 gulls, 15 expressed virus antigen in their tissues. Virus antigen expression was limited to epithelial cells of intestine and cloacal bursa. No histological lesions were detected in association with virus antigen expression. Our findings show that LPAIV replication in the intestinal tract of black-headed gulls is mainly a superficial infection in absence of detectable lesions, as determined recently for natural LPAIV infection in free-living mallards (Anas platyrhynchos). These findings imply that LPAIV in black-headed gulls has adapted to minimal pathogenicity to its host and that potentially the primary transmission route is faecal-oral.
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http://dx.doi.org/10.1080/03079457.2012.744447DOI Listing
December 2012

Ecological factors driving avian influenza virus dynamics in Spanish wetland ecosystems.

PLoS One 2012 12;7(11):e46418. Epub 2012 Nov 12.

Instituto de Investigación en Recursos Cinegéticos, UCLM-JCCM-CSIC, Ciudad Real, Spain.

Studies exploring the ecological interactions between avian influenza viruses (AIV), natural hosts and the environment are scarce. Most work has focused on viral survival and transmission under laboratory conditions and through mathematical modelling. However, more integrated studies performed under field conditions are required to validate these results. In this study, we combined information on bird community, environmental factors and viral epidemiology to assess the contribution of biotic and abiotic factors in the occurrence of low pathogenic AIV in Spanish wetlands. For that purpose, seven locations in five different wetlands were studied during two years (2007-2009), including seven sampling visits by location. In each survey, fresh faeces (n = 4578) of wild birds and water samples were collected for viral detection. Also, the vegetation structure, water physical properties of wetlands, climatic conditions and wild bird community composition were determined. An overall AIV prevalence of 1.7%±0.4 was detected in faecal samples with important fluctuations among seasons and locations. Twenty-six AIV were isolated from the 78 RRT-PCR positive samples and eight different haemagglutinines and five neuraminidases were identified, being the combination H3N8 the most frequent. Variation partitioning procedures identified the combination of space and time variables as the most important pure factor - independently to other factors - explaining the variation in AIV prevalence (36.8%), followed by meteorological factor (21.5%) and wild bird community composition/vegetation structure (21.1%). These results contribute to the understanding of AIV ecological drivers in Spanish ecosystems and provide useful guidelines for AIV risk assessment identifying potential hotspots of AIV activity.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0046418PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3495955PMC
May 2013

Natural Bagaza virus infection in game birds in southern Spain.

Vet Res 2012 Sep 11;43:65. Epub 2012 Sep 11.

Instituto de Investigación en Recursos Cinegéticos IREC, (CSIC-UCLM-JCCM), Ronda de Toledo s/n, 13071, Ciudad Real, Spain.

In late summer 2010 a mosquito born flavivirus not previously reported in Europe called Bagaza virus (BAGV) caused high mortality in red-legged partridges (Alectoris rufa) and ring-necked pheasants (Phasianus colchicus). We studied clinical findings, lesions and viral antigen distribution in naturally BAGV infected game birds in order to understand the apparently higher impact on red-legged partridges. The disease induced neurologic signs in the two galliform species and, to a lesser extent, in common wood pigeons (Columba palumbus). In red-legged partridges infection by BAGV caused severe haemosiderosis in the liver and spleen that was absent in pheasants and less evident in common wood pigeons. Also, BAGV antigen was present in vascular endothelium in multiple organs in red-legged partridges, and in the spleen in common wood pigeons, while in ring-necked pheasants it was only detected in neurons and glial cells in the brain. These findings indicate tropism of BAGV for endothelial cells and a severe haemolytic process in red-legged partridges in addition to the central nervous lesions that were found in all three species.
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http://dx.doi.org/10.1186/1297-9716-43-65DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3483237PMC
September 2012

Occurrence of avian pathogenic Escherichia coli and antimicrobial-resistant E. coli in red-legged partridges (Alectoris rufa): sanitary concerns of farming.

Avian Pathol 2012 ;41(4):337-44

Instituto de Investigación en Recursos Cinegéticos IREC, Ronda de Toledo s/n (CSIC-UCLM-JCCM), 13005, Ciudad Real, Spain.

Red-legged partridges (Alectoris rufa) are a significant part of the culture, diet and income for many people in central and southern Spain. Due to declining populations in the wild, intensive farming is common and 4 million juvenile partridges are released each autumn. Intensive management and high densities result in high prevalence of enteric disease and the use of antimicrobials as preventive measures on partridge farms and prior to restocking in the wild. We determined the occurrence of avian pathogenic Escherichia coli (APEC), and screened phenotypic resistance of E. coli against enrofloxacin, gentamicin and cefotaxim in farmed, restocked and wild partridges. Prevalence of APEC in farmed and restocked red-legged partridges was significantly higher than in natural populations. Phenotypic resistance against both gentamicin and enrofloxacin was significantly more frequent in farmed (75%) and restocked (43%) partridges than in wild partridges, while most E. coli isolated from natural populations were susceptible to all three antimicrobials tested (65%). This indicates that farmed and restocked partridges carry APEC that could be a reason for disease outbreaks on farms, and that E. coli carried by farmed and restocked partridges can acquire resistance to frequently used antimicrobials, thus being a concern for the environment, wild birds and consumers. Management in farms and restocking procedures may create a hazard not only for spreading APEC, but also as a potential source of resistant E. coli in the environment.
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http://dx.doi.org/10.1080/03079457.2012.687101DOI Listing
August 2014