Occurrence and genetic variability of Phlebotomus papatasi in an urban area of southern Italy

<p>Abstract</p> <p>Background</p> <p>A phlebotomine sand fly was noticed in the second floor of an old building in a highly urbanized area of southern Italy. A short-term entomological survey was carried out in the subsequent weeks to this event, allowing the collection of additional phlebotomine sand flies that were later identified as <it>Phlebotomus papatasi</it>. We assessed the genetic variability among <it>P</it>. <it>papatasi </it>sequences obtained in this study and those available from Italy using a mitochondrial DNA (mtDNA) fragment (from <it>cytochrome b </it>gene to NADH1) and the internal transcribed spacer 2 (ITS2) as genetic markers.</p> <p>Results</p> <p>From 9 June to 19 July, eight males and seven females (two blood-fed) of <it>P</it>. <it>papatasi </it>were collected in the old town of Bari (southern Italy). The insects were found near the bed and in the bathroom and potential blood sources (e.g., pigeons and dogs) for them were common in the neighbourhood. Again, five females of <it>P</it>. <it>papatasi </it>collected in Valenzano, another urban area in the province of Bari, were also identified and included in the genetic study. The mtDNA sequences (945 bp) obtained from Bari and Valenzano were identical except for a single transition (T ↔ C) at the 793 nucleotide residue. Pairwise comparison of the last 440 bp of the mtDNA fragment analyzed herein with other sequences of <it>P. papatasi </it>from Italy revealed a nucleotide variation ranging from 0.2 to 1.3%. Three ITS2 sequence types were detected within specimens collected in Valenzano, one of them identical to that from Bari. Pairwise comparison of ITS2 sequences of <it>P. papatasi </it>from Italy revealed a nucleotide variation up to 1.8%.</p> <p>Conclusions</p> <p>This study reports the occurrence of <it>P</it>. <it>papatasi </it>in an urban area of southern Italy and shows a low nucleotide difference among ITS2 and mtDNA sequences of this species available from Italy. The presence of <it>P</it>. <it>papatasi </it>in urban areas might represent a risk for human health, particularly for the potential transmission of sandfly fever viruses.</p

Exon-intron structure and sequence variation of the calreticulin gene among Rhipicephalus sanguineus group ticks

Background: Calreticulin proteins (CRTs) are important components of tick saliva, which is involved in the blood meal success, pathogen transmission and host allergic responses. The characterization of the genes encoding for salivary proteins, such as CRTs, is pivotal to understand the mechanisms of tick-host interaction during blood meal and to develop tick control strategies based on their inhibition. In hard ticks, crt genes were shown to have only one intron with conserved position among species. In this study we investigated the exon-intron structure and variation of the crt gene in Rhipicephalus spp. ticks in order to assess the crt exon-intron structure and the potential utility of crt gene as a molecular marker. Methods: We sequenced the exon-intron region of crt gene in ticks belonging to so-called tropical and temperate lineages of Rhipicephalus sanguineus (sensu lato), Rhipicephalus sp. I, Rhipicephalus sp. III, Rhipicephalus sp. IV, R. guilhoni, R. muhsamae and R. turanicus. Genetic divergence and phylogenetic relationships between the sequences obtained were estimated. Results: All individuals belonging to the tropical lineage of R. sanguineus (s. l.), R. guilhoni, R. muhsamae, R. turanicus, Rhipicephalus sp. III and Rhipicephalus sp. IV analysed showed crt intron-present alleles. However, both crt intron-present and intron-absent alleles were found in Rhipicephalus sp. I and the temperate lineage of R. sanguineus (s. l.), showing the occurrence of an intraspecific intron presence-absence polymorphism. Phylogenetic relationships among the crt intron-present sequences showed distinct lineages for all taxa, with the tropical and temperate lineages of R. sanguineus (s. l.) being more closely related to each other. Conclusions: We expanded previous studies about the characterization of crt gene in hard ticks. Our results highlighted a previously overlooked variation in the crt structure among Rhipicephalus spp., and among hard ticks in general. Notably, the intron presence/absence polymorphism observed herein can be a candidate study-system to investigate the early stages of intron gain/loss before fixation at species level and some debated questions about intron evolution. Finally, the sequence variation observed supports the suitability of the crt gene for molecular recognition of Rhipicephalus spp. and for phylogenetic studies in association with other markers

Molecular detection of Leishmania spp. in dogs and a cat from Doha, Qatar

Research Areas: Parasitology ;Tropical MedicineBackground: Canine and feline leishmanioses are increasingly reported worldwide and represent a threat to both animal and human health. Despite their relevance, data about leishmanioses in companion animals in the east-central part of the Arabian Peninsula are unavailable. Therefore, we investigated the occurrence of Leishmania spp. in dogs and cats from Qatar. Methods: From March 2016 to May 2018, 199 pets (120 dogs and 79 cats) living in Doha or its outskirts were included in this study. From each animal a blood sample was collected and tested for Leishmania spp. by quantitative real-time PCR. Results: Out of the 199 animals, four (2.0%) were positive for Leishmania spp., including three dogs (2.5%) and one cat (1.3%). Conclusions: All positive animals were born in Qatar and had not travelled overseas, suggesting that infection was locally acquired. Considering the occurrence of Leishmania spp. and its potential impact on the health of animals and humans, it is crucial to increase scientific knowledge in order to plan screening and regular prophylaxis against sand fly vectors to reduce the risk of infection.info:eu-repo/semantics/publishedVersio

Biological compatibility between two temperate lineages of brown dog ticks, Rhipicephalus sanguineus (sensu lato)

Background: The brown dog tick Rhipicephalus sanguineus (sensu stricto) is reputed to be the most widespread tick of domestic dogs worldwide and has also been implicated in the transmission of many pathogens to dogs and humans. For more than two centuries, Rh. sanguineus (s.s.) was regarded as a single taxon, even considering its poor original description and the inexistence of a type specimen. However, genetic and crossbreeding experiments have indicated the existence of at least two distinct taxa within this name: the so-called "temperate" and "tropical" lineages of Rh. sanguineus (sensu lato). Recent genetic studies have also demonstrated the existence of additional lineages of Rh. sanguineus (s.l.) in Europe and Asia. Herein, we assessed the biological compatibility between two lineages of Rh. sanguineus (s.l.) found in southern Europe, namely Rhipicephalus sp. I (from Italy) and Rhipicephalus sp. II (from Portugal). Methods: Ticks morphologically identified as Rh. sanguineus (s.l.) were collected in southern Portugal and southern Italy. Tick colonies were established and crossbreeding experiments conducted. Morphological, biological and genetic analyses were conducted. Results: Crossbreeding experiments confirmed that ticks from the two studied lineages were able to mate and generate fertile hybrids. Hybrid adult ticks always presented the same genotype of the mother, confirming maternal inheritance of mtDNA. However, larvae and nymphs originated from Rhipicephalus sp. I females presented mtDNA genotype of either Rhipicephalus sp. I or Rhipicephalus sp. II, suggesting the occurrence of paternal inheritance or mitochondrial heteroplasmy. While biologically compatible, these lineages are distinct genetically and phenotypically. Conclusions: The temperate lineages of Rh. sanguineus (s.l.) studied herein are biologically compatible and genetic data obtained from both pure and hybrid lines indicate the occurrence of paternal inheritance or mitochondrial heteroplasmy. This study opens new research avenues and raises question regarding the usefulness of genetic data and crossbreeding experiments as criteria for the definition of cryptic species in ticks

the eyeworm thelazia callipaeda in portugal current status of infection in pets and wild mammals and case report in a beech marten martes foina

Abstract Ocular thelaziosis is caused by nematodes of the genus Thelazia (Spirurida, Thelaziidae), which inhabit the surface of the eyes and associated tissues. Thelazia callipaeda affects a range of mammal species, including humans, and in the last two decades has been reported in multiple European countries, being classified as an emergent vector-borne pathogen. In Portugal T. callipaeda is endemic in north-eastern areas, where it has been reported in domestic dogs, cats, red foxes (Vulpes vulpes) and wild European rabbits (Oryctolagus cuniculus). We report, for the first time, T. callipaeda in a beech marten (Martes foina) from Portugal and highlight the presence of haplotype 1 as the only one found in Europe, irrespective of the host species and geographical area of provenience

Clinical case presentation and a review of the literature of canine onchocercosis by Onchocerca lupi in the United States

Background: Onchocerca lupi, a filarioid of zoonotic concern, infects dogs and cats causing ocular lesions of different degrees, from minor to severe. However, infected animals do not always display overt clinical signs, rendering the diagnosis of the infection obscure to the majority of veterinarians. Canine onchocercosis has been reported in the Old World and the information on its occurrence in the United States, as well as its pathogenesis and clinical management is still meagre. This study reports on the largest case series of O. lupi infection from the United States and reviews previous cases of canine onchocercosis in this country. Methods: Information on the clinical history of a series of eight cases of O. lupi infection in dogs diagnosed in Minnesota, New Mexico, Colorado and Florida, from 2011 to 2014, was obtained from clinical records provided the veterinary practitioners. Nematodes were morphologically identified at species level and genetically analyzed. Results: All dogs displayed a similar clinical presentation, including subconjunctival and episcleral nodules, which were surgically removed. Each dog was subjected to post-operative therapy. Whitish filaria-like parasites were morphologically and molecularly identified as O. lupi. Conclusions: This study confirms that O. lupi is endemic in the United States, indicating that the distribution of the infection is probably wider than previously thought. With effect, further studies are urgently needed in order to improve the diagnosis and to assess the efficacy of therapeutic protocols, targeting the parasite itself and/or its endosymbionts

Zoonotic Abbreviata caucasica in wild chimpanzees (Pan troglodytes verus) from Senegal

Abbreviata caucasica (syn. Physaloptera mordens) has been reported in human and various non-human primates including great apes. The identification of this nematode is seldom performed and relies on egg characterization at the coproscopy, in the absence of any molecular tool. Following the recovery of two adult females of A. caucasica from the feces of wild Senegalese chimpanzees, morphometric characteristics were reported and new data on the width of the esophagus (0.268-0.287 mm) and on the cuticle structure (0.70-0.122 mm) were provided. The molecular characterization of a set of mitochondrial (cox1, 16S rRNA, 12S rRNA) and nuclear (18S rRNA and ITS2) partial genes was performed. Our phylogenetic analysis indicates for the first time that A. caucasica is monophyletic with Physaloptera species. A novel molecular tool was developed for the routine diagnosis of A. caucasica and the surveillance of Nematoda infestations. An A. caucasica-specific qPCR targeting the 12S gene was assessed. The assay was able to detect up to 1.13 × 10−3 eggs/g of fecal matter irrespective of its consistency, with an efficiency of 101.8% and a perfect adjustment (R2 = 0.99). The infection rate by A. caucasica in the chimpanzee fecal samples was 52.08%. Only 6.19% of the environmental samples were positive for nematode DNA and any for A. caucasica. Our findings indicate the need for further studies to clarify the epidemiology, circulation, life cycle, and possible pathological effects of this infestation using the molecular tool herein developed

National quality assurance guidelines for architectural education - help or hindrance in curriculum development and innovation

This paper will outline the nature of a national guideline for quality assurance and standards in architectural education. It will also report on the application of the guidelines in testing the curriculum of the Bachelors and Masters programmes of the Cork Centre for Architectural Education. Finally it will evaluate the robustness of the guidelines in accommodating curriculum change in the context of technology change, by considering the specific case of incorporating into the curriculum, education and training for architects and students aimed at assisting them in contributing to the development of the "intelligent" building. The author will draw upon recent experience serving on an Expert Committee charged with drawing up guidelines for the Quality Assurance in architectural education in the Republic of Ireland. This work was undertaken at the behest of HETAC, the Higher Education and Training Awards Council, a body established by the Irish government with responsibility for the maintenance of quality and standards in many parts of the third level (post secondary school) education. The preparation of guidelines on quality and standards in architectural education was timely given some significant changes in the context of professional practice and architectural education in Ireland. The country introduced legislation in 2007, in the Building Act, which protects the title of "architect" and its usage. The professional body, the Royal Institute of the Architects of Ireland, has been appointed as the agency responsible for the registration of architects within the terms of the new law, and for the accreditation of their training and education. This responsibility also extends to control at the national level, of entry to the profession within the European Union, as defined in "Professional Qualifications" (EU Directive 2005/36). Furthermore, after a prolonged period of unprecedented growth in the construction sector and growth in demand for architectural services, lasting till 2007, the number of architectural schools in the country grew from 2003, from just two, long-established schools in the capital city, Dublin, to a total of five in 2006, with the new schools being distributed across the country. The new HETAC document attempts to identify the abilities, skills, and knowledge that graduates should achieve at different levels in their architectural education. These guidelines are used as an index to test the curriculum at the author's own school, the Cork Centre for Architectural Education. The results will be used to evaluate the utility of the HETAC document in evaluating curricula, and in supporting the maintenance of quality, and facilitating innovation. To investigate the latter issues of innovation and curriculum development, the author will take as a case study, the extent to which the new guidelines encourage, or inhibit the incorporation of curriculum change driven by technology change. Specifically it will examine the application of recommendations for the education of architects and architectural students being developed within the NEMBES project, an Irish Government funded (PRTLI-IV), multi-disciplinary project aimed at developing a Centre of Excellence in Cork for the application of embedded technology in the built environment. This curriculum change is intended to enable architects to contribute to the development of "intelligent" environments through utilising electronic sensors and actuators enabling performance and configurationally changes in built enclosures, in response to environmental parameters and occupant behaviour

Chronic polyarthritis associated to Cercopithifilaria bainae infection in a dog.

Despite the widespread distribution of Cercopithifilaria bainae among canine and tick populations worldwide, this filarioid is currently considered of 'minor importance' in veterinary medicine, particularly when compared to related filarioids, such as Dirofilaria immitis and Dirofilaria repens. To date, only a single case of dermatological alterations possibly associated to infection by C. bainae had been reported in a dog. In the present study, we describe the first case of systemic alterations associated to C. bainae infection in a dog suffering from diffused chronic polyarthritis. The animal had a previous history of reluctance to move and stiff gait and displayed multiple joint pain during manipulation of limbs. No biochemical, haematological and X-ray alterations were detected; microfilariae were observed in the synovial fluids collected from the joints. In spite of the morphological and molecular identification of these microfilariae as C. bainae, the dog did not respond to multiple microfilaricidal treatments with milbemicyn oxyme. The potential role of C. bainae in the pathogenesis of this clinical condition is discussed. Given the potential pathogenicity of this parasite, improved knowledge of this little known tick-borne nematode is warranted in order to assist the development of novel and effective treatment strategies.This is the author accepted manuscript. The final version can be found on the publisher's website at: http://www.sciencedirect.com/science/article/pii/S0304401714003744# © 2014 Published by Elsevier B.V

Vertical transmission of Anaplasma platys and Leishmania infantum in dogs during the first half of gestation.

BACKGROUND: Leishmania infantum is a canine zoonotic vector-borne protozoan pathogen transmitted by phlebotomine sand flies, whereas Anaplasma platys is a bacterium most likely transmitted by ticks. While vertical transmission of L. infantum from pregnant bitches to their offspring has been documented, thus far no studies have explored the possibility of vertical transmission of A. platys in dogs. This study investigated the occurrence of vertical transmission of L. infantum and A. platys in sheltered dogs during the first half of gestation, in an area of southern Italy characterised by a high incidence of infection by both pathogens. METHODS: The study population included 20 bitches (n = 10 pregnant, at 25-35 days of pregnancy; n = 10 non-pregnant), all subjected to ovariohysterectomy, which were examined for the presence of L. infantum and A. platys via cytological screening of bone marrow and whole blood samples. Infection by L. infantum and A. platys was also tested by immunofluorescence antibody test (IFAT) and quantitative real-time PCR (qPCR) targeting both pathogens. Selected tissue samples (n = 210) collected during surgical procedures from bitches and foetuses (n = 20) were assessed for the presence of L. infantum and A. platys by qPCR targeting a fragment of the kinetoplast minicircle DNA (kDNA) and the 16S rRNA gene, respectively. RESULTS: Leishmania infantum DNA was not amplified from either uteri or ovaries from pregnant bitches or foetal tissue samples, whereas a subset of ovarian (n = 2) and uterine (n = 4) tissue samples from non-pregnant bitches were infected, with parasite loads of up to 3.09 × 10 and 7.51 parasite/PCR reaction, respectively. Conversely, uterine (n = 10) and ovarian (n = 8) tissues from both pregnant and non-pregnant bitches, together with a subset (n = 5) of foetal tissue samples were qPCR positive for A. platys. Leishmania infantum and A. platys nucleic acids were amplified from two uteri from non-pregnant bitches, with parasite loads of up to 2.32 × 10(-3) and 2.05 parasite/per PCR reaction, respectively. CONCLUSIONS: Results from this study suggest that, in contrast to L. infantum, A. platys can be transmitted from pregnant dogs to their offspring during the first half of gestation. This hypothesis remains to be verified, for instance via direct observation of parasites in postpartum foetal tissues.The paper has been sponsored by Bayer Animal Health in the framework of the 11th CVBD World Forum Symposium