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Projeto de investigação
PHENOTYPIC AND GENETIC CARACTERIZATION OF CULICOIDES DIPTERA: CERATOPOGONIDAE IN PORTUGAL AND COMPARISON OF THE EFFECT OF INSECTICIDE MOLECULES IN THEIR CONTROL
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Phenotypic and genetic characterization of Culicoides (DIPTERA: CERATOPOGONIDAE) in Portugal and comparison of the effect of pyrethroid insecticides in their control
Publication . Ramilo, David Wilson Russo; Sampaio, Isabel Maria Soares Pereira da Fonseca de; Curdi, Javier Lucientes
Culicoides genus is of major importance in animal and human health since hematophagous females are vectors of several pathogens, like viruses (Bluetongue, African Horse Sickness and Schmallenberg) and filarial nematodes, among others. As a consequence, female biting midges are responsible for huge economical losses worldwide. A deeper knowledge of Culicoides fauna present in each country and their ecological preferences is required, so different control strategies can be applied efficiently.
The present work was based on Culicoides species captures during the National Entomologic Surveillance Program for Bluetongue disease in mainland Portugal and Azores and Madeira archipelagos (2005-2013), using miniature CDC light traps, and in 2015, with OVI traps, in the framework of VectorNet European network.
Biting midges were evaluated phenotypically and genetically, showing the variation of Culicoides species in Portuguese territory. Twenty-two Culicoides species were mentioned for the first time in Portugal, including C. dewulfi and C. montanus in mainland Portugal and species of Obsoletus group in the islands of Azores archipelago where they were never reported, as well as a description of Culicoides paradoxalis, a new species for science. Moreover, a detailed study focused the morphology, genetics and ecology of Obsoletus group, showing that the distribution of those species was unequal in mainland Portugal. Plus, a redefinition of the 3rd palpus segment length/width ratio and spermathecae size intervals, aiming Obsoletus group species identification, together with the reference of anatomical aberrations in these species, was performed, allowing the reduction of errors during midges studies. An identification key for all known Portuguese midges was also created.
In this thesis, elaboration of risk assessment maps based on the association of some abiotic variables with the occurrence of C. imicola, C. pulicaris, C. punctatus, C. newsteadi and species from Obsoletus group in mainland Portugal were also performed.
Finally, evaluation of C. imicola morphological modifications in sensorial organs localized in the 3rd palpus segment, used for host detection, was performed after an assay with pyrethroid insecticides (permethrin and deltamethrin) at different concentrations, showing complete destruction of sensorial organs with probable feeding implications.
The results presented in this scientific work contributed to a better knowledge of Culicoides genus in Portugal, being some of them of worldwide relevance.
Uropathogenic Proteus mirabilis and Klebsiella pneumoniae in companion animals : molecular epidemiology, antimicrobial resistance and zoonotic potential
Publication . Marques, Cátia; Pomba, Maria Constança Matias Ferreira
Urinary tract infections (UTI) are frequently diagnosed in companion animals and the increase
in antimicrobial resistance leads to therapeutic limitations and public health concerns.
The study of the geographic distribution of antimicrobial resistance in bacteria (n= 22 256)
causing UTI in companion animals from 14 European countries showed that, in 2012-2013,
the frequency of Escherichia coli and Proteus mirabilis antimicrobial resistance in Southern
countries (Portugal, Spain, Italy, Greece) was significantly higher than in Northern countries
(Denmark, Sweden). In a retrospective study conducted in Portugal (Lisbon), the antimicrobial
resistance of clinical Enterobacteriaceae from companion animals with UTI increased
significantly over 16 years (1999-2014; P < 0.001). Bacteria from companion animals with UTI
harboured important antimicrobial resistance mechanisms and belonged to high-risk clonal
lineages, namely third-generation cephalosporin (3GC)-resistant E. coli O25b:H4-B2-ST131,
CC23 and ST648, methicilin-resistant Staphylococcus aureus CC5, methicilin-resistant
Staphylococcus epidermidis CC5, high-level gentamicin-resistant Enterococcus faecalis CC6
and ampicillin-resistant Enterococcus faecium CC17.
The blaCTX-M-15 gene was disseminated among 3GC-resistant K. pneumoniae from companion
animals and humans with UTI. Most K. pneumoniae from companion animals where 3GC,
multidrug-resistant (MDR) and belonged to the high-risk clonal lineage ST15. K. pneumoniae
high-risk clonal lineages ST11, ST37 and ST147 were also detected in companion animals.
3GC-resistance in P. mirabilis from companion animals with UTI was associated with the
presence of blaCMY-2, which increased significantly over time. A high number of PFGE clusters
(43.6%, n = 17/39) included P. mirabilis strains from companion animals and humans.
Gut colonisation by K. pneumoniae and P. mirabilis was detected in healthy dogs and humans;
however, none of the strains was MDR. K. pneumoniae faecal strains from dogs belonged to
ST17, ST188, ST252, ST281, ST423, ST1093, ST1241, ST3398 and ST3399. Remarkably,
two colonised dogs were found to shared PFGE undistinguishable K. pneumoniae strains with
one co-living human. P. mirabilis gut colonisation was significantly higher in dogs (P = 0.0329).
One human/dog and one dog/dog pair shared PFGE undistinguishable P. mirabilis strains.
The antimicrobial resistance frequencies reported in these studies support the need to
implement antimicrobial stewardship programmes in veterinary medicine. The detection of
MDR high-risk clonal lineages causing UTI in companion animals and the similarities detected
in K. pneumoniae and P. mirabilis from companion animals and humans raises public health
concerns and highlights the need for a One Health approach.
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Fundação para a Ciência e a Tecnologia
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SFRH/BD/77268/2011