Susceptibility in vitro of canine methicillin-resistant and -susceptible staphylococcal isolates to fusidic acid, chlorhexidine and miconazole: opportunities for topical therapy of canine superficial pyoderma

OBJECTIVES: Increasing multidrug resistance amongst canine pathogenic staphylococci has renewed interest in topical antibacterial therapy for skin infections in the context of responsible veterinary prescribing. We therefore determined the activity in vitro of three clinically relevant topical agents and synergism between two of them against Staphylococcus pseudintermedius and Staphylococcus aureus. METHODS: The MICs of fusidic acid (n = 199), chlorhexidine (n = 198), miconazole (n = 198) and a 1:1 combination of miconazole/chlorhexidine (n = 198) were determined for canine isolates [50 MRSA and 49 methicillin-resistant S. pseudintermedius (MRSP), 50 MSSA and 50 methicillin-susceptible S. pseudintermedius (MSSP)] collected from the UK and Germany using an agar dilution method (CLSI VET01-A4). Fractional inhibitory concentration (FIC) indices were calculated to assess the interaction of miconazole with chlorhexidine. RESULTS: MICs of each drug/combination were significantly (P < 0.0005) higher for S. aureus when compared with S. pseudintermedius. Most strains (n = 172) had an MIC of fusidic acid of ≤0.03 mg/L (MIC ≥64 mg/L, n = 5 MRSA). All strains had MICs of chlorhexidine of 0.5–4 mg/L, except for one MRSA (MIC = 8 mg/L). All but four strains had MICs of miconazole of 1–4 mg/L (MIC = 16 mg/L, n = 3; MIC = 256 mg/L, n = 1). Miconazole/chlorhexidine (1:1 ratio) had a synergistic effect against 49/50 MRSA, 31/50 MSSA, 12/49 MRSP and 23/49 MSSP. CONCLUSIONS: Since the majority of these staphylococci, including methicillin-resistant isolates, had MICs that should be readily exceeded by topical skin application of these agents, their therapeutic efficacy for canine superficial pyoderma should be assessed. The synergistic interaction shown in vitro supports further clinical evaluation of miconazole/chlorhexidine combination therapy for staphylococcal infection

A new routing protocol for ad hoc wireless networks design, implementation and performance evaluation

A collection of mobile nodes can form a multi-hop radio network with a dynamic topology and without the need for any infrastructure such as base stations or wired network. Such a Mobile Ad Hoc Networks (MANETs) maintain their structure and connectivity in a decentralised and distributed fashion. Each mobile node acts as both a router for other nodes traffic, as well as a source of traffic of its own In this thesis we develop and present a new hybrid routing protocol called Multipath Distance Vector Zone Routing Protocol, which is referred to as MDVZRP. In MDVZRP we assume that all the routes in the routing table are active and usable at any time, unless the node received or discovered a broken link. There is no need to periodically update the routing tables, therefore reducing the periodic update messages and hence reducing the control traffic in the entire network. The protocol guarantees loop freedom and alternative disjoint paths. Routes are immediately available within each routing zone. For destinations outside the zone, MDVZRP employs a route discovery technique known as routing information on demand. Once the node is informed by either the MAC layer or itself that it should discover the non- reachable nodes, MDVZRP adopts a new technique. First, we discuss the Ad Hoc networks and routing in general, then the motivation of MDVZRP regarding the nodes‟ flat view, and the selection and acquisition of multipath getting and selection. Furthermore, we describe the stages of MDVZRP and the protocol routing process with examples. The performance of MDVZRP is then evaluated to determine its operating parameters, and also to investigate its performance in a range of different scenarios. Finally, MDVZRP is compared with DSDV and AODV ordinary routing protocols (standard) delivering CBR traffic. Simulation results show that MDVZRP gives a better performance than DSDV in all circumstances, it is also better than AODV in most of the scenarios, especially at low mobility

MRSA carrying <i>mecC</i> in captive mara

This is the final published version. It first appeared at http://jac.oxfordjournals.org/content/early/2015/02/24/jac.dkv024.full.Objectives: To characterize the staphylococcal cassette chromosome mec (SCCmec), virulence and antimicrobial\ud susceptibility of Staphylococcus aureus ST130 isolated from mara (Dolichotis patagonum), a large rodent species\ud native to South America and kept in captivity at Copenhagen Zoo.\ud Methods: The presence of mecCwas confirmed by PCR in 15 S. aureus ST130 isolated from mara during a previous\ud study. WGS was performed on two randomly selected isolates to characterize their genomes with respect to\ud SCCmec, virulence and resistance gene content. Antimicrobial susceptibility was tested using commercial\ud broth microdilution tests.\ud Results: All the isolates belonged to spa type t528 ST130 and carried mecC. Based on WGS, mecC was 100%\ud identical to the prototype sequence of S. aureus strain LGA251. The sequence of SCCmec type XI in the mara\ud isolates had 23 SNPs compared with the one described in LGA251. The two sequenced strains harboured a set\ud of virulence factors and other genomic features previously observed in ST130. Both strains carried norA as the\ud only putative antimicrobial resistance gene in addition to mecC.\ud Conclusions: Our findings support the notion that a genetically conserved mecC-carryingMRSA ST130 clone iswidespread\ud in a variety of unrelated hosts in Denmark. Since the mara at Copenhagen Zoo have limited contact with\ud humans and other animal species, it remains unclear whether mara are natural hosts of ST130 or acquired this lineage\ud from unknown sources. The broad host range of MRSA ST130 supports its designation as a generalist lineage.This study was supported by internal funding (C. E.-G., A. M. and L. G.) and by the Medical Research Council (E. M. H. and M. A. H.; G1001787/1)

Evidence for the evolutionary steps leading to mecA-mediated ß-lactam resistance in staphylococci

The epidemiologically most important mechanism of antibiotic resistance in Staphylococcus aureus is associated with mecA–an acquired gene encoding an extra penicillin-binding protein (PBP2a) with low affinity to virtually all β-lactams. The introduction of mecA into the S. aureus chromosome has led to the emergence of methicillin-resistant S. aureus (MRSA) pandemics, responsible for high rates of mortality worldwide. Nonetheless, little is known regarding the origin and evolution of mecA. Different mecA homologues have been identified in species belonging to the Staphylococcus sciuri group representing the most primitive staphylococci. In this study we aimed to identify evolutionary steps linking these mecA precursors to the β-lactam resistance gene mecA and the resistance phenotype. We sequenced genomes of 106 S. sciuri, S. vitulinus and S. fleurettii strains and determined their oxacillin susceptibility profiles. Single-nucleotide polymorphism (SNP) analysis of the core genome was performed to assess the genetic relatedness of the isolates. Phylogenetic analysis of the mecA gene homologues and promoters was achieved through nucleotide/amino acid sequence alignments and mutation rates were estimated using a Bayesian analysis. Furthermore, the predicted structure of mecA homologue-encoded PBPs of oxacillin-susceptible and -resistant strains were compared. We showed for the first time that oxacillin resistance in the S. sciuri group has emerged multiple times and by a variety of different mechanisms. Development of resistance occurred through several steps including structural diversification of the non-binding domain of native PBPs; changes in the promoters of mecA homologues; acquisition of SCCmec and adaptation of the bacterial genetic background. Moreover, our results suggest that it was exposure to β-lactams in human-created environments that has driven evolution of native PBPs towards a resistance determinant. The evolution of β-lactam resistance in staphylococci highlights the numerous resources available to bacteria to adapt to the selective pressure of antibiotics

Characterization, mechanism of action and optimization of activity of a novel peptide-peptoid hybrid against bacterial pathogens involved in canine skin infections

Integumentary infections like pyoderma represent the main reason for antimicrobial prescription in dogs. Staphylococcus pseudintermedius and Pseudomonas aeruginosa are frequently identified in these infections, and both bacteria are challenging to combat due to resistance. To avoid use of important human antibiotics for treatment of animal infections there is a pressing need for novel narrow-spectrum antimicrobial agents in veterinary medicine. Herein, we characterize the in vitro activity of the novel peptide-peptoid hybrid B1 against canine isolates of S. pseudintermedius and P. aeruginosa. B1 showed potent minimum inhibitory concentrations (MICs) against canine S. pseudintermedius and P. aeruginosa isolates as well rapid killing kinetics. B1 was found to disrupt the membrane integrity and affect cell-wall synthesis in methicillin-resistant S. pseudintermedius (MRSP). We generated 28 analogues of B1, showing comparable haemolysis and MICs against MRSP and P. aeruginosa. The most active analogues (23, 26) and B1 were tested against a collection of clinical isolates from canine, of which only B1 showed potent activity. Our best compound 26, displayed activity against P. aeruginosa and S. pseudintermedius, but not the closely related S. aureus. This work shows that design of target-specific veterinary antimicrobial agents is possible, even species within a genus, and deserves further exploration

Prioritization of Companion Animal Transmissible Diseases for Policy Intervention in Europe

A number of papers have been published on the prioritization of transmissible diseases in farm animals and wildlife, based either on semiquantitative or truly quantitative methods, but there is no published literature on the prioritization of transmissible diseases in companion animals. In this study, available epidemiological data for diseases transmissible from companion animals to man were analysed with the aim of developing a procedure suitable for their prioritization within a European framework. A new method and its associated questionnaire and scoring system were designed based on methods described by the World Organisation for Animal Health (OIE). Modifications were applied to allow for the paucity of specific information on companion animal transmissible diseases. The OIE method was also adapted to the subject and to the regional scope of the interprofessional network addressing zoonotic diseases transmitted via companion animals in Europe: the Companion Animals multisectoriaL interprofessionaL Interdisciplinary Strategic Think tank On zoonoses (CALLISTO). Adaptations were made based on information collected from expert groups on viral, bacterial and parasitic diseases using a structured questionnaire, in which all questions were closed-ended. The expert groups were asked to select the most appropriate answer for each question taking into account the relevance and reliability of the data available in the scientific literature. Subsequently, the scoring of the answers obtained for each disease covered by the questionnaire was analysed to obtain two final overall scores, one for human health impact and one for agricultural economic impact. The adapted method was then applied to select the 15 most important pathogens (five for each pathogen group: viral, bacterial and parasitic) on the basis of their overall impact on public health and agriculture. The result of the prioritization exercise was a joint priority list (available at www.callistoproject.eu) of relevant pathogens according to these two criteria. As the scope of CALLISTO was comprehensive in terms of geographical area, animal species involved and impact of the diseases, the list of prioritized diseases had to accommodate the realities in different European countries and the differences in biology and animalehuman relationships in a wide range of species including cats and dogs, pet pigs and sheep as well as captive reptiles. The methodology presented in this paper can be used to generate accurate priority lists according to narrower and more specific objectives

Genome and Plasmid Sequences of Escherichia coli KV7, an Extended-Spectrum β-Lactamase Isolate Derived from Feces of a Healthy Pig.

We present single-contig assemblies for Escherichia coli strain KV7 (serotype O27, phylogenetic group D) and its six plasmids, isolated from a healthy pig, as determined by PacBio RS II and Illumina MiSeq sequencing. The chromosome of 4,997,475 bp and G+C content of 50.75% harbored 4,540 protein-encoding genes