Prevalence of extended-spectrum beta-lactamase-producing Escherichia coli isolates in faecal samples of broilers

Seventy-six faecal samples were obtained from broilers at slaughterhouse level in Portugal. Samples were inoculated on cefotaxime-supplemented Levine agar plates. Cefotaxime-resistant Escherichia coli isolates were recovered from 32 samples (42.1%), obtaining a total of 34 E. coli isolates (one or two isolates per sample). Susceptibility to 16 antibiotics was studied by disk diffusion method, and 85% of the isolates presented a phenotype of multi-resistance that included antimicrobial agents of at least four different families. Extended-spectrum-beta-lactamases (ESBL) of the TEM and CTX-M groups were detected in 31 ESBL-positive E. coli isolates. Twenty-six isolates harboured the bla_TEM-52 gene and two of them also harboured bla_TEM-1b. The bla_CTX-M-14 gene was identified in three isolates (in association with bla_TEM-1b in one of them), and bla_CTX-M-32 was demonstrated in two additional isolates. Three of the 34 cefotaxime-resistant isolates (9%) did not produce ESBLs, and two of them presented mutations at positions −42 (C → T), −18 (G → A), −1 (C → T), and +58(C → T) of the promoter/attenuator region of ampC gene. tet(A) and/or tet(B) genes were detected in all 34 tetracycline-resistant isolates, aadA in all 26 streptomycin-resistant isolates; cmlA in 3 of 6 chloramphenicol-resistant isolates, and aac(3)-II or aac(3)-I + aac(3)-IV genes in all 4 gentamicin-resistant isolates. Different combinations of sul1, sul2 and sul3 genes were demonstrated among the 22 trimethoprim–sulfamethoxazole-resistant isolates. Amino acid changes in GyrA and ParC proteins were identified in all 18 ciprofloxacin-resistant isolates. The results of this study indicate that the intestinal tract of healthy poultry is a reservoir of ESBL-positive E. coli isolates.     

Risk factors for canine leishmaniasis in an endemic Mediterranean region

Human visceral leishmaniasis is an emergent/re-emergent parasitic zoonotic disease in Europe caused by Leishmania infantum, with domestic dog as its main reservoir host. This study presents the results of a canine epidemiological survey in a mediterranean region where human and canine leishmaniasis (CanL) are endemic - Portugal. The main goal was to identify risk factors, which can be relevant for Leishmania infection control. The national survey was carried out in January 2009 with a screening of 3974 dogs from all 18 districts of mainland Portugal. Direct Agglutination Test was used for the detection of anti-Leishmania antibodies in canine blood. An overall CanL true prevalence of 6.31% was observed. Apparent prevalence at district level ranged from 0.88% to 16.16%, with the highest prevalence in the interior regions. Identified risk factors for positivity were: dogs of 2 years and older (adjusted odds ratio OR=5.39); spending exclusively/most of the time outdoors (OR=2.51); origin from the interior of Portugal in comparison to littoral/coast districts (OR=2.51); not having long fur (OR=2.03); and being pure exotic (OR=1.67). The results confirm the leishmaniasis endemicity in Portugal and the dynamic character of prevalence as new foci emerged and old foci lost their importance. The dog's age, fur size, district and living outdoors as opposed to indoors were more important than dog breeds and insecticide treatment in the transmission of Leishmania infection. The future of CanL prevention and control rely on an integrated approach involving veterinarians, dog owners and health authorities in order to reduce the canine infection risk and consequently, the human zoonotic visceral leishmaniasis.     

Do domestic cats impose an unsustainable harvest on urban bird populations?

We assessed the impact of domestic cats on population persistence of native and exotic urban bird populations using a model adjusted for habitat-specific catch rates, cat ownership and hunting activity data. GPS-derived home ranges of 32 cats and resource selection indices demonstrated the degree of penetration and preference for native vegetation fragments. Owners reported on prey brought back by 144 domestic cats in Dunedin, New Zealand, during 12 months. One third of cats never brought back prey, and 21% returned more than one item/month. Cats brought back a mean of 13.4 prey items/year (median = 4), with cats aged <1 year returning more prey than older cats. Birds were the most common prey, followed by rodents. Although cats penetrated adjacent vegetation fragments they did not catch more birds and preferred garden habitat, suggesting that predation pressure may be reduced in fragments. Cat home range size appears to be constrained by cat density while the number of birds caught depends on the density of available prey. Estimates of city-wide catch for six bird species were either more than total urban population size estimates or close to lower confidence intervals. Modelling of three species indicated low likelihood of population persistence with cat predation. The observed persistence of these prey species suggests a meta-population structure with urban populations acting as sinks with source populations located on the city fringe.     

Heuristic algorithms vs. linear programs for designing efficient conservation reserve networks: Evaluation of solution optimality and processing time

Systematic approaches to efficient reserve network design often make use of one of two types of site selection algorithm; linear programs or heuristic algorithms. Unlike with linear programs, heuristic algorithms have been demonstrated to yield suboptimal networks in that more sites are selected in order to meet conservation goals than may be necessary or fewer features are captured than is possible. Although the degree of suboptimality is not known when using heuristics, some researchers have suggested that it is not significant in most cases and that heuristics are preferred since they are more flexible and can yield a solution more quickly. Using eight binary datasets, we demonstrate that suboptimality of numbers of sites selected and biodiversity features protected can occur to various degrees depending on the dataset, the model design, and the type of heuristic applied, and that processing time is not dramatically different between optimal and heuristic algorithms. In choosing an algorithm, the degree of suboptimality may not always be as important to planners as the perception that optimal solvers have feasibility issues, and therefore heuristic algorithms might continue to be a popular tool for conservation planning. We conclude that for many datasets, feasibility of optimal algorithms should not be a concern and that the value of heuristic results can be greatly improved by using optimal algorithms to determine the degree of suboptimality of the results.     

Identification and typing of Vagococcus salmoninarum using genomic and proteomic techniques

This study reports on the characterization of Vagococcus salmoninarum using phenotypic, serological, antigenic, genetic and proteomic methods. All strains of V. salmoninarum were resistant to most of the antimicrobials tested, and only 10% of strains were sensitive to florfenicol. Serological analysis demonstrated a high antigenic homogeneity within the species. No cross‐reaction was detected with other fish pathogenic species causing streptococcosis (Lactococcus garvieae, Streptococcus parauberis, Streptococcus iniae, Streptococcus agalactiae, Carnobacterium maltaromaticum) using serum against V. salmoninarum CECT 5810. Electrophoretic analysis of cell surface proteins and immunoblot supported the antigenic homogeneity within V. salmoninarum strains. Moreover, limited diversity was detected using genomic (RAPD, ERIC‐PCR and REP‐PCR) and MALDI‐TOF‐MS analyses. The phenotypic, genomic and proteomic methods tested allowed the rapid differentiation of V. salmoninarum from the other species causing streptococcosis. However, MALDI‐TOF‐MS is the most promising method for typing and characterization of V. salmoninarum.     

High‐throughput identification and quantification of Vagococcus salmoninarum by SYBR Green I‐based real‐time PCR combined with melting curve analysis

This work describes a primer pair and a high‐throughput SYBR Green I‐based real‐time PCR protocol combined with melting curve analysis for identification and quantification of Vagococcus salmoninarum in bacterial cultures and infected fish tissues. The 16S rRNA gene was selected for the design of the primer pair (SalF and SalR). The sensitivity and specificity of this primer pair were compared with other previously designed for conventional PCR. Although both primer pairs showed 100% specificity using pure bacterial cultures or DNA extracted from bacteria or fish tissues, the primer pairs designed in this study showed the highest sensitivity with a detection limit of 0.034 × 10⁰ amplicon copies per assay (equivalent to 2 × 10^?11 ng/µl, C_q value of 30.49 ± 1.71). The developed qPCR protocol allowed the detection of V. salmoninarum in non‐lethal and lethal fish samples with detection levels of 0.17 × 10⁰ gene copies in tissues artificially infected and 0.02 × 10⁰ in tissues of fish experimentally infected with V. salmoninarum. The high sensitivity of the developed method suggests that it could be considered as a useful tool for diagnosis of vagococcosis and the detection of V. salmoninarum in asymptomatic or carrier fish.     

Avian pathogenic <Emphasis Type="Italic">Escherichia coli</Emphasis> bind fibronectin and laminin

Avian colisepticemia frequently occurs after respiratory tract damage, the primary site for infection allows bacteria to encounter an exposed basement membrane, where laminin and fibronectin are important components. We investigated the ability of an isolate of avian pathogenic Escherichia coli to bind fibronectin and laminin. Using Far-western dot blot analysis, we demonstrated the ability of this microorganism to bind basement membrane proteins fibronectin and laminin. Results from an ELISA-based approach indicate that the binding to these membrane proteins was bacterial-dose dependent. Furthermore, two specific E. coli polypeptides, of 32 kDa and 130 kDa, reacted with laminin and fibronectin, respectively. Further evaluation of these potential bacterial adhesins may provide insights into the pathogenesis of colibacillosis.