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Background: Aedes albopictus has spread during the last few decades all over the world. This has increased significantly the risk of exotic arbovirus transmission (e.g. chikungunya, dengue, and Zika) also in temperate areas, as demonstrated by the Chikungunya 2007 and 2017 outbreaks in northeastern and central Italy. Insecticides are an important tool for limiting the circulation of these mosquito-borne viruses. The aim of the present study was to address the gap in current knowledge of pyrethroid insecticide resistance of European Ae. albopictus populations, focusing on populations from Italy, Albania and Greece., Results: Bioassays for resistance to permethrin (0.75%), α-cypermethrin (0.05%) or deltamethrin (0.05%) were performed according to World Health Organization (WHO) protocols and showed reduced susceptibility (<90% mortality) of some Italian populations to permethrin and α-cypermethrin, but not to deltamethrin., Conclusion: This study reports the first evidence of resistance to pyrethroids in adult Italian Ae. albopictus populations. Results refer to the season preceding the Chikungunya 2017 outbreak in central Italy and highlight the need to increase efforts to monitor the spread of insecticide resistance and the need to develop strategies to limit the spread of insecticide resistance, particularly in areas where extensive treatments have been carried out to contain disease outbreaks. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

The immature immune system of cats in their paediatric age (i.e., first six months of life) favours the establishment of infectious respiratory tract diseases mainly caused by well recognized viruses and bacteria species. Conversely, lungworm infections are less investigated during respiratory disorders in kittens. In the last decade, Troglostrongylus brevior has been found affecting the respiratory tract of cats, along with the better-known Aelurostrongylus abstrusus. Given the scant data available on the epidemiology of feline troglostrongylosis, faecal samples from 575 domestic animals living in three Italian municipalities (i.e., Bari, Messina and Siena) were screened for lungworm infection by Baermann and molecular tecniques. Animals were grouped according to their age as ≤6 months (i.e., paediatric patients), 6-24 months, or >24 months. Paediatric cats were further sub-divided in infant (2-6 weeks), weanling (6-12 weeks) and juvenile (3-6 months). Of the 575 animals tested, 241 (42.0%) were younger than 6 months, 188 (33%) were 6-24-month-old and 146 (25%) were older than 24 months. Lungworm infection was diagnosed in 84 (14.6%) of the examined cats. Of the 49 (20.3%) paediatric animals positive for lungworms, T. brevior was the nematode species most frequently diagnosed (n = 44; 89.8%), followed by A. abstrusus (n = 2; 4.1%), and three cats (6.1%) were co-infected by both species. The diagnosis of T. brevior infection was significantly associated with animals aging ≤6 months (18.2%; P < 0.01) than elder cats. Indeed, the prevalence of infection by T. brevior decreased in animals aging 6-24 months (3.2%) being not detected in cats older than two years. Results of this study indicate that paediatric cats are at higher risk of T. brevior infection compared to adults (P < 0.01)., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Cats that have outdoor access are highly exposed to ticks, fleas, mites and flying insects, though the risk to become infested by arthropods is less perceived in cats than in dogs. This has resulted in fewer treatment and prevention options being available for cats than for dogs. A collar containing a combination of 10% imidacloprid and 4.5% flumethrin (Seresto ® , Bayer Animal Health) is available for cats and licensed with claims against ticks and fleas for 7-8 months. Following the assessment of the efficacy of the collar against Leishmania infantum infection in privately owned cats living in the Aeolian archipelago, herein we report the efficacy of the collar in the treatment and prevention of tick and flea infestations in the same population of cats over a period of one year of observation. At the inclusion day (Study Day 0, SD 0), cats were visited and examined for ectoparasites (i.e., flea combing and tick thumb counts) and allocated to group 1 (G1; n=104; cats treated with Seresto ® collar) or group 2 (G2; n=100; untreated controls) and further checked at SDs 210, 270 and 360 (study closure). At SD 0, G1 and G2 had a comparable percentage of cats infested by fleas (45.2% and 49.0%; χ 2 =0.164; P=0.6859) and ticks (6.7% and 14.0%; χ 2 =2.946; P=0.0861). The number of cats infested by fleas was reduced in G1, being 8.3%, 0% and 3.8% on SDs 210, 270 and 360, respectively, resulting in efficacies against fleas of 79.4%, 100% and 93.6% on SDs 210, 270 and 360. None of the cats in G1 was found infested by ticks after the application of the collar, whereas in G2 ticks were observed on 15.7%, 4.8%, 17.5% of the cats at the different follow up visits, leading to an overall efficacy against ticks of 100%. A total of 375 ectoparasites were collected from cats, being 249 fleas (six Ctenocephalides canis, 240 Ctenocephalides felis and three Nosopsyllus fasciatus) and 126 ticks (87 Ixodes ventalloi and 39 Rhipicephalus pusillus). Field data gathered herein confirm a high efficacy of the collar in the prevention of tick and flea infestations on cats. This is of great importance both for the primary role of fleas and ticks as blood feeding parasites and, more importantly, because of their role as vectors of pathogens causing diseases of veterinary and medical importance., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

Introduction: Aelurostrongylus abstrusus is a well-known nematode affecting the respiratory system of felids worldwide. Snails and slugs act as intermediate hosts of this parasite, whereas rodents, birds and reptiles may serve as paratenic hosts. Periplaneta americana, the American brown cockroach, shares the same habitat and ecological features (e.g. nocturnal activity) with both snails and cats. The aim of this study was to evaluate the capability of P. americana to maintain alive A. abstrusus third stage larvae (L3s) after artificial inoculation., Material and Methods: Twenty-five specimens of P. americana were infected with 100 A. abstrusus L3s collected from experimentally infected Cornu aspersum snails, whereas five specimens were used as control group. After the infection, cockroaches were maintained in individual plastic boxes until dissection for the presence of L3s at 1 (T1), 5 (T5), 10 (T10), 15 (T15), and 20 (T20) days post-infection., Results: Except for T15, alive A. abstrusus L3s (n = 63) were found at all time-points, being 26, 19, 16 and 2 L3s retrieved at T1, T5, T10 and T20, respectively. Eleven (17.4%) L3s were found within the digestive tract, 10 (15.9%) in other-than-digestive organs and 42 (66.7%) in the exoskeleton and associated tissues. Nine out of the twenty-five experimentally inoculated cockroaches (36%) died soon after the artificial infection (T1), while in the control group, two out of the five (40%) died before the end of the study (T15) with no difference in the mortality rate between groups., Discussion: Results of this study suggest that P. americana could act as a paratenic host of A. abstrusus. Periplaneta americana cockroaches, have a ubiquitous distribution and may be preyed by cats, representing a potential source of infection to cats living in endemic areas., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Light traps are one of the most common attractive method for the collection of nocturnal insects. Although light traps are generally referred to as "CDC light traps", different models, equipped with incandescent or UV lamps, have been developed. A new light trap, named Laika trap 3.0, equipped with LED lamps and featured with a light and handy design, has been recently proposed into the market. In this study we tested and compared the capture performances of this new trap with those of a classical light trap model under field conditions. From May to November 2013, a Laika trap and a classical light trap were placed biweekly in an area endemic for sand flies. A total of 256 sand fly specimens, belonging to 3 species (Sergentomyia minuta, Phlebotomus perniciosus, Phlebotomus neglectus) were collected during the study period. The Laika trap captured 126 phlebotomine sand flies: P. perniciosus (n=38); S. minuta (n=88), a similar number of specimens (130) and the same species were captured by classical light trap which collected also 3 specimens of P. neglectus. No significant differences in the capture efficiency at each day of trapping, neither in the number of species or in the sex of sand flies were observed. According to results of this study, the Laika trap may be a valid alternative to classical light trap models especially when handy design and low power consumption are key factors in field studies., (Copyright © 2017 Elsevier B.V. All rights reserved.)

With the exception of Aelurostrongylus abstrusus, feline lungworms have been poorly studied. Information on their distribution is patchy and mostly limited to case reports. In this study, the occurrence of feline lungworms and co-infecting gastrointestinal parasites has been investigated in 12 European countries (i.e. Austria, Belgium, Bulgaria, France, Greece, Hungary, Italy, Portugal, Romania, Spain, Switzerland and the United Kingdom). An average of 10 domestic cats, with regular outdoor access, was sampled each month for 12months, and freshly passed faeces were collected. Stools were processed using a McMaster assay and a quantitative Baermann-Wetzel method. Animals positive for lungworms and/or gastrointestinal parasites were treated with a formulation containing fipronil, (S)-methoprene, eprinomectin, and praziquantel (Broadline®, Merial), and re-sampled 28days post-treatment. The association between lungworm infection and risk factors was analysed using statistical medians/means and the efficacy of the treatment against each lungworm species was assessed. Of 1990 cats sampled, 613 (30.8%) were positive for at least one parasite, while 210 (10.6%) were infected by lungworms. The prevalence of lungworm infection varied between the sampled sites, with the highest recorded in Bulgaria (35.8%) and the lowest in Switzerland (0.8%). None of the cats from Austria or the United Kingdom were infected by lungworms. Aelurostrongylus abstrusus was the species most frequently detected (78.1%), followed by Troglostrongylus brevior (19.5%), Eucoleus aerophilus (14.8%) and Oslerus rostratus (3.8%). The overall efficacy of the treatment was 99% for A. abstrusus and 100% for T. brevior, O. rostratus and E. aerophilus. Data presented provide a comprehensive account of the diagnosis, epidemiology and treatment of feline lungworms in Europe, as well as of the occurrence of co-infections by gastrointestinal parasites., (Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Leishmaniosis caused by Leishmania infantum is one of the most important vector-borne diseases affecting animals and humans worldwide. Dogs are considered main reservoirs of the zoonotic forms, though in the last years the role of cats as reservoirs has been increasingly investigated. Feline leishmaniosis (FeL) occurs in endemic areas and no specific preventive measures have been investigated so far. In this study the efficacy of a 10% imidacloprid/4.5% flumethrin polymer matrix collar, licensed for tick and flea prevention, has been assessed against FeL in a longitudinal study on 204 privately owned cats from the Aeolian islands (Sicily), an area highly endemic for the disease. From March to May 2015 [Study Day 0 (SD 0)], cats negative for FeL were collared (G1, n = 104) or left untreated (G2, n = 100). Diagnosis consisted of serology and qPCR on blood and conjunctival swabs, which were collected at baseline (SD 0) and at the end of the study (SD 360). Interim clinical examinations were performed on SD 210 (when collars were replaced in G1) and SD 270., Results: Of the 159 cats which completed the study, 5 in G1 and 20 in G2 were positive for L. infantum infection, in at least one of the diagnostic tests leading to a yearly crude incidence of 6.3% and 25.0% in G1 and G2, respectively (P = 0.0026). This translates into an efficacy of the collar of 75.0% in preventing feline Leishmania infection. The collar was generally well tolerated with no systemic adverse reactions and few local skin reactions were observed in the application area in four out of 104 treated cats (3.8%)., Conclusions: The 10% imidacloprid/4.5% flumethrin collar significantly reduced the risk of L. infantum infection in cats. To our knowledge, this is the first study in which a preventative strategy against feline Leishmania infection is assessed under natural conditions. These findings close a gap in veterinary medicine, in that they confirm this collar as a tool in reducing the risk of Leishmania infection in cats. Such a preventative tool could contribute to the reduction of the risk of the disease in animal and in human populations when included in integrated leishmaniosis control programmes.

Vector-borne diseases (VBDs) are prevalently investigated in dogs. Studies on feline VBDs are scant, though feline leishmaniosis (FeL) is increasingly recognised as a disease of cats in endemic areas. Comprehensive investigations on the distribution of VBDs in populations of cats and dogs living in relatively small geographical areas, such as islands, are currently lacking. In this study the prevalence of Leishmania infantum and other VBD pathogens was assessed in cohorts of cats and dogs living in the Aeolian Islands. Autochthonous animals (330 cats and 263 dogs) of different age and sex were sampled. Blood and conjunctival samples were collected from cats and dogs for serological and molecular testing. Eighty-five (25.8%) cats were positive for L. infantum, 13 (3.9%) for Bartonella spp. and 1 (0.3%) for Hepatozoon felis. One-hundred and ten dogs (41.8%) were positive for L. infantum and three (1.1%) for Hepatozoon canis. The incidence of L. infantum infection in cats positive after one season of exposure to sand fly was 14.7%. Leishmania infantum prevalence and year incidence were higher in dogs than in cats (p=0.0001 and p=0.0003, respectively). Thirty-four cats (10.3%) scored positive for ticks (mean intensity rate of infestation, 2.03±1.4), which were identified to the species level as Ixodes ventalloi and Rhipicephalus pusillus. Conversely, Rhipicephalus sanguineus sensu lato (s.l.) was the only species identified in dogs (10.6%). A larger prevalence of infestation by Ctenocephalides felis was recorded in cats (n=91; 27.6%) than in dogs (n=33; 12.5%) (p=0.0001). In addition, one female Nosopsyllus fasciatus (syn. Ceratophyllus fasciatus) and one male Spilopsyllus cuniculi were also identified in flea-infected cats. VBDs are endemic in the Aeolian Islands being L. infantum the most prevalent vector-borne pathogen circulating between cats and dogs. The overall seroprevalence of FeL herein recorded is higher than that assessed, only by IFAT, in populations of cats in Greece and in Spain. Because L. infantum and VBDs are more commonly associated with dogs, the recognition of cats as hosts of different vector-borne pathogens is of paramount importance towards a better management of these diseases in both animals and humans., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

Despite their medical and veterinary importance, some tick species are so poorly studied, that their role within pathogen vector transmission cycles is difficult to assess. The tick Ixodes ventalloi is one such species, and its biology and phylogenetic status remain an issue of debate. In the present study, specimens of adult I. ventalloi (n = 65 females; n = 31 males) infesting cats in the Lipari Island (Aeolian archipelago, Sicily, southern Italy) were characterized morphologically and molecularly, the latter based on mitochondrial 16S rRNA and cytochrome c oxidase subunit 1 (cox1) genes. The genetic data and phylogenetic analyses for both mitochondrial genes suggest the existence of two distinct genogroups. The ecological and epidemiological significance of the genetic structure within the I. ventalloi endemic population remains to be determined. The results highlight the need for further analysis of this tick species, including whole mitochondrial genome sequencing and crossbreeding studies, which will be pivotal to complement features of its status as a vector of pathogens.

Afoxolaner (AFX) plus milbemycin oxime (MO) combination chewable tablets (NexGard Spectra®, Merial) were evaluated for safety and efficacy against naturally acquired nematode infections in domestic dogs in a multi-centre, positive control, blinded field study using a randomized block design based on the order of presentation for allocation. In total, 408 dogs confirmed positive for naturally acquired infections of intestinal nematodes by pre-treatment faecal examination were studied in ten countries in Europe (Albania, Austria, Bulgaria, France, Germany, Hungary, Italy, Lithuania, Romania and Slovakia). Pre-treatment faecal examination revealed Toxocara, Toxascaris, hookworm, Trichuris and/or Capillaria nematode infections in 134, 30, 223, 155 and 14 dogs, respectively. Dogs were allocated to one of two treatment groups in a ratio of 1, AFX + MO chewables (≥2.5 mg AFX + ≥0.5 mg MO per kg body weight, according to dose bands; 207 dogs), and 1, MO plus praziquantel (PRZ) chewables (Milbemax®, Novartis; ≥0.5 mg MO + ≥5 mg PRZ per kg body weight, according to the manufacturer's instructions; 201 dogs) and treated once. For evaluation of efficacy based on reduction of faecal nematode egg counts, two faecal samples, one collected prior to treatment and one collected 9 to 21 days after treatment, were examined using modified McMaster techniques. For evaluation of systemic safety, dogs were examined by a veterinarian before treatment administration and at study end, and dog owners observed the health status of their dogs until the end of the study and reported any abnormal observation. For dogs treated with AFX + MO chewables, the efficacy was 99.7, 99.7, 97.2, 99.7 and 99.7 % for Toxocara, Toxascaris, hookworm, Trichuris and Capillaria, respectively; and the efficacy was 99.5, 99.4, 94.3, 99.9 and 98.0 %, respectively, for the MO + PRZ-treated dogs (p ≤ 0.002 for all nematodes and both treatments). For Toxocara, hookworm and Trichuris, non-inferiority analysis demonstrated that the efficacy of AFX + MO chewable tablets was equal to or better than that of MO + PRZ. In spite that both treatments were ≥98 % efficacious against Toxascaris and Capillaria, a hypothesis of non-inferiority for both genera could not be established due to the low number of dogs infected with these parasites. No treatment-related adverse experiences were observed throughout the study. For both treatments, all dogs were given a systemic safety score of 'excellent' apart from one dog in each treatment group which received a score of 'acceptable'. AFX + MO combination chewables were shown to be safe and demonstrated a high level of efficacy when administered once to dogs infected with a broad range of parasitic nematodes under field conditions.

This study investigated the efficacy of two collars for the treatment and prevention of flea infestations. Additionally the effect of these collars on the incidence of Leishmania infantum infection as compared with a group of vaccinated dogs was evaluated. A total of 224 young dogs from private animal shelters were enrolled in April/May into four groups: G1, 55 dogs treated with 10% imidacloprid + 4.5% flumethrin collar (Seresto, Bayer Animal Health); G2, 60 dogs treated with 4% deltamethrin collar (Scalibor protector band, MSD Animal Health); G3, 54 dogs vaccinated with CaniLeish (Virbac Animal Health); and G4, 55 dogs left non-treated as controls. Dogs were followed up at days 120 (September), 210 (December), and 360 (April-May). At those time points, clinical assessments, ectoparasite counts and blood, bone marrow and skin samples, to detect the presence of L. infantum, were performed. The efficacy of Seresto in protecting dogs from flea infestation was 100% (P < 0.01) on day 120 and 210, while animals treated with Scalibor showed a prevalence of the infestation ranging from 23.3% to 33.3% on day 120 and 210, respectively. At the end of the study, the incidence of L. infantum infection in collared dogs-based on animals being positive in any of the tests-was 5.5% in Seresto-treated dogs and 20% in Scalibor-treated dogs, resulting in overall efficacy of prevention of 88.3% for Seresto and 61.8% for Scalibor. No statistical difference was detected in L. infantum positive dogs for bone marrow PCR and/or cytology at day 360 between the CaniLeish (15.4%) and non-treated control dogs (10.0%). Both collars proved to be effective (P < 0.01) in preventing L. infantum infection throughout one transmission season, whereas no significant difference was recorded in the frequency of active infections between dogs vaccinated with CaniLeish and control dogs, emphasizing the importance of using repellent/insecticide actives as a priority measure for protection against canine leishmaniosis., Competing Interests: I have read the journal's policy and the authors of this manuscript have the following competing interests: The study was supported by Bayer Animal Health GmbH (Germany) and Dr. Dorothee Stanneck is an employee of Bayer Animal Health GmbH, Leverkusen, Germany. The study was monitored by Arccoblu s.r.l (Italy); Dr. Fabrizio Solari Basano and Roberto Nazzari are employees of Arcoblu s.r.l. There are no patents, products in development or marketed products to declare.

The laboratory maintenance of parasitic life cycles is crucial to support research in many fields of parasitology. The land snail Helix aspersa (syn. Cornu aspersum), an intermediate host of feline lungworms Aelurostrongylus abstrusus and Troglostrongylus brevior, is adopted to produce infective stages of those nematodes in laboratory condition. The aim of this study was to compare the most common methods of experimental infection of H. aspersa with first stage larvae (L1) of A. abstrusus (i.e., contact of the snail foot with the L1) with the injection of these larvae in the foot of the snail, instrumental to reduce the infection time and to maximize the output of third-stage larvae (L3). Three groups (i.e., A, B, C) of 15 H. aspersa snails were infected with L1 of A. abstrusus (n=250 for each snail), whereas a fourth group (group D) was not infected (control). Snails were individually placed for 48h on a microfilm containing L1 (group A), on a potato slice previously irrigated with a suspension of L1 (group B), or they were inoculated by injection of L1 in the posterior-ventral portion of the foot (group C). Eighteen days after the infection all snails were analyzed and tissues were digested to recover L3. No difference in mortality rate was recorded among snail groups and the mean number of retrieved L3 was significantly larger in group C (71.5±52.9) compared to group B (38.2±44.9; p=0.0161) and group A (19±23.3; p<0.0001). The injection of A. abstrusus L1 in the foot of H. aspersa proved to be a fast, easy to apply and effective method, resulting in the largest number of infective larvae retrieved., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Red fox (Vulpes vulpes) is considered as reservoir of different cardiorespiratory parasites of veterinary and medical importance. Since data on cardiorespiratory parasites in foxes in Bosnia and Herzegovina are still lacking, the aims of the present study were to (i) investigate the prevalence and geographical distribution of these parasites, (ii) determine genetic diversity of detected parasite species, and (iii) to estimate the role of foxes in the transmission cycle to companion animals and humans. Four species, morphologically and molecularly identified as Eucoleus boehmi (64.6%; 51/79), Eucoleus aerophilus (69.7%; 154/221), Crenosoma vulpis (45.7%; 101/221) and Linguatula serrata (1.3%; 1/79) were retrieved from nasal cavity and lungs in 184 (83.3%) animals. The occurrence of heartworms, Angiostrongylus vasorum and Dirofilaria immitis was not detected by necropsy or PCR. Furthermore, three distinct haplotypes of E. aerophilus (I, III, XV) and two of C. vulpis (I, II) previously reported in pet animals and wild carnivores were confirmed in this study. A new haplotype of C. vulpis (designated as haplotype V) was also identified based on 12S rRNA gene for the first time. The present study indicates a high prevalence and wide distribution of nasal and lung nematodes in fox population in Bosnia and Herzegovina, and supports the existence of transmission patterns between wildlife and pet animals., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: The Comunity Health Centre (CHC) primary care model is a team-based health care delivery model intended to provide comprehensive and continuous medical care to patients within a defined community. The CHC, Case della Salute in Italian, model was introduced in the Emilia-Romagna Region in 2010., Methods: We present updated data on the implementation on the CHC Case della Salute primary care model in the Emilia-Romagna Region., Results: There are 67 operating CHCs in Emilia-Romagna (update March 2015); 26 small (39%), 24 medium (36%) and 17 large (25%). Since 2011 the number of operating CHCs has increased by 60%, reaching 55% of the target planned CHCs (n. = 122). There is, on average, one running CHC per 66.524 inhabitants. 16% of total general practitioners (GPs) and 8.4% of total family paediatricians working in Emilia-Romagna have their practice in CHCs. CHCs offer primary and specialist integrated care, prevention services, health education and social care., Discussion: Although preliminary results suggest CHCs have fostered primary care's quality and efficiency, more research is needed to assess their impact on improving clinical, social and economic outcomes.

Wildcats are endangered felid species living in Europe, Asia, and Africa. Regrettably, scientific information on parasites of wildcats is particularly meager and they often rely on data gained by necropsies of a small number of animals. In the present study, scat collection was used to assess the parasite spectrum of European wildcats living in the Etna Park (Sicily, Italy). Scat collection was performed from May to September 2010 by weekly walking four transects for a total of 391 km. Samples were then analyzed by flotation and sedimentation techniques to investigate wildcat parasitic fauna. A total of 121 scats of wildcats were collected, and parasitic forms (i.e., oocysts, eggs, and larvae) were retrieved in 110 (90.9 %) of the samples. Parasites found were Physaloptera sp. (52.1 %), tapeworms (45.5 %), Toxocara cati (43.8 %), Eucoleus aerophilus (27.3 %), Ancylostoma sp. (22.3 %), Troglostrongylus brevior (15.7 %), trematodes (9.9 %), Isospora felis (4.1 %), Cylicospirura sp. (1.7 %), and Acanthocephala (0.8 %). The prevalence of endoparasitic infections herein recorded is similar to that described in other studies conducted using necropsy technique. The species richness of parasites found in the present survey, with a total of nine helminths and one protozoon, is the highest ever reported for wildcat in Europe. Scat collection and examination are reliable and rapid non-invasive tools which can be used in a systematic survey design to study the parasite spectrum of wildcat as well as that of other endangered wild species.

Ovitraps are regarded as a reliable system to monitor Aedes albopictus dynamics. However, the dimensions of the oviposition substrate are not standardized, and no studies have investigated which should be the most effective sizes. In this study, the effect of paddle sizes in tiger mosquito egg collection was evaluated. Egg count and density on the wide surfaces and margins of different-sized oviposition substrates have been evaluated in two studies (A and B). In study A, a total of 29,995 Ae. albopictus eggs was counted in 250 classic oviposition substrates. Eggs were found on both wide surfaces (53.1%) and margins (46.9%). Egg density was significantly larger in margins compared to wide surfaces. Overall in study B, 983 Ae. albopictus eggs were collected. According to paddle sizes, 51.8% of eggs were on large and 48.2% on small paddles. Mean egg density of wide surfaces was significantly larger in small paddles (0.25 eggs/cm(2) ) compared to large paddles (0.06 eggs/cm(2) ). Results indicate that wider oviposition substrates do not mean larger number of Ae. albopictus eggs. Indeed, on paddles four times thinner than others, the number of eggs counted was not statistically different. These findings suggest that small paddles may be routinely employed in ovitraps, thus allowing savings of materials and money., (© 2015 The Society for Vector Ecology.)

Over the last few decades, ecological factors, combined with everchanging landscapes mainly linked to human activities (e.g. encroachment and tourism) have contributed to modifications in the transmission of parasitic diseases from domestic to wildlife carnivores and vice versa. In the first of this two-part review article, we have provided an account of diseases caused by protozoan parasites characterised by a two-way transmission route between domestic and wild carnivore species. In this second and final part, we focus our attention on parasitic diseases caused by helminth and arthropod parasites shared between domestic and wild canids and felids in Europe. While a complete understanding of the biology, ecology and epidemiology of these parasites is particularly challenging to achieve, especially given the complexity of the environments in which these diseases perpetuate, advancements in current knowledge of transmission routes is crucial to provide policy-makers with clear indications on strategies to reduce the impact of these diseases on changing ecosystems., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Over the last few decades, the world has witnessed radical changes in climate, landscape, and ecosystems. These events, together with other factors such as increasing illegal wildlife trade and changing human behaviour towards wildlife, are resulting into thinning boundaries between wild canids and felids and their domestic counterparts. As a consequence, the epidemiology of diseases caused by a number of infectious agents is undergoing profound readjustements, as pathogens adapt to new hosts and environments. Therefore, there is a risk for diseases of wildlife to spread to domestic carnivores and vice versa, and for zoonotic agents to emerge or re-emerge in human populations. Hence, the identification of the hazards arising from the co-habitation of these species is critical in order to plan and develop adequate control strategies against these pathogens. In the first of this two-part article, we review the role that wild canids and felids may play in the transmission of protozoa and arthropod-borne agents to dogs and cats in Europe, and provide an account of how current and future progress in our understanding of the ecology and epidemiology of parasites, as well as of host-parasite interactions, can assist efforts aimed at controlling parasite transmission., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Snail-borne lungworms exert an enormous toll on the health and welfare of animals and humans. Of these parasites, Aelurostrongylus abstrusus and Troglostrongylus brevior affect the respiratory tract of felids. These lungworms share both the ecological niche and the species of snail (Helix aspersa) acting as intermediate host. Recently, the ability of H. aspersa to shed infective third-stage larvae (L3s) of A. abstrusus and T. brevior in the environment has been demonstrated, matching previous knowledge of mode of transmission of zoonotic lungworms. Here, we evaluated, for the first time, the ability of A. abstrusus and T. brevior L3s to infect new, susceptible snail hosts following their release from experimentally infected molluscs, and refer to this novel route of parasite transmission as intermediesis. The implications of snail-to-snail transmission in the epidemiology of snail-borne diseases are also discussed.

Background: Cyclospora is a protistan parasite that causes enteritis in several species of animals including humans. The aim of this study was to investigate the presence of Cyclospora in captive non-human primates., Methods: A total of 119 faecal samples from Pan troglodytes, Macaca sylvanus, Cercopithecus cephus, Erythrocebus patas, Chlorocebus aethiops and Macaca fascicularis from a wildlife animal rescue center as well as from Macaca fascicularis from an experimental primate research center were tested for the presence of Cyclospora by quantitative real-time PCR (qPCR) and single-strand conformation polymorphism (SSCP) analysis., Results: Cyclospora was detected in three Pan troglodytes (13.6%) and nine (9.3%) Macaca fascicularis., Conclusions: The present study represents the first record of Cyclospora in captive primates in Europe, suggesting the presence of Cyclospora cayetanensis, which is transmissible to humans.

The increasing reports of Aelurostrongylus abstrusus infection and the new information on Troglostrongylus brevior have spurred the interest of the scientific community towards the research of pharmaceutical compounds effective against both pathogens. A novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel (Broadline®, Merial) has been released for the treatment of a variety of feline parasitic infections. The present study reports the efficacy of this spot-on in treating cats naturally infected by feline lungworms. Client owned cats (n=191) were enrolled from three geographical areas of Italy and faecal samples were examined by floatation and Baermann techniques. Twenty-three individuals were positive for L1 of A. abstrusus (n=18) or T. brevior (n=3) or for both species (n=2) and they were topically treated with Broadline®. Seventeen of them were also concomitantly infected by other parasites. Four weeks after treatment, faecal samples were collected and examined to assess the efficacy of a single administration of the product. Based on lungworm larvae counts, the efficacy of the treatment was 90.5% or 100% for A. abstrusus or T. brevior, respectively. Cats released significantly lower amounts of lungworm larvae after treatment compared to pre-treatment (p<0.0001). All but three cats were negative for other nematodes after treatment and all cats recovered from respiratory signs. Results of this study indicate that a single administration of the topical combination fipronil, (S)-methoprene, eprinomectin and praziquantel is effective and safe for the treatment of A. abstrusus and/or T. brevior infections in cats living under field conditions., (Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Gastropod-borne parasites may cause debilitating clinical conditions in animals and humans following the consumption of infected intermediate or paratenic hosts. However, the ingestion of fresh vegetables contaminated by snail mucus and/or water has also been proposed as a source of the infection for some zoonotic metastrongyloids (e.g., Angiostrongylus cantonensis). In the meantime, the feline lungworms Aelurostrongylus abstrusus and Troglostrongylus brevior are increasingly spreading among cat populations, along with their gastropod intermediate hosts. The aim of this study was to assess the potential of alternative transmission pathways for A. abstrusus and T. brevior L3 via the mucus of infected Helix aspersa snails and the water where gastropods died. In addition, the histological examination of snail specimens provided information on the larval localization and inflammatory reactions in the intermediate host., Methodology/principal Findings: Twenty-four specimens of H. aspersa received ~500 L1 of A. abstrusus and T. brevior, and were assigned to six study groups. Snails were subjected to different mechanical and chemical stimuli throughout 20 days in order to elicit the production of mucus. At the end of the study, gastropods were submerged in tap water and the sediment was observed for lungworm larvae for three consecutive days. Finally, snails were artificially digested and recovered larvae were counted and morphologically and molecularly identified. The anatomical localization of A. abstrusus and T. brevior larvae within snail tissues was investigated by histology. L3 were detected in the snail mucus (i.e., 37 A. abstrusus and 19 T. brevior) and in the sediment of submerged specimens (172 A. abstrusus and 39 T. brevior). Following the artificial digestion of H. aspersa snails, a mean number of 127.8 A. abstrusus and 60.3 T. brevior larvae were recovered. The number of snail sections positive for A. abstrusus was higher than those for T. brevior., Conclusions: Results of this study indicate that A. abstrusus and T. brevior infective L3 are shed in the mucus of H. aspersa or in water where infected gastropods had died submerged. Both elimination pathways may represent alternative route(s) of environmental contamination and source of the infection for these nematodes under field conditions and may significantly affect the epidemiology of feline lungworms. Considering that snails may act as intermediate hosts for other metastrongyloid species, the environmental contamination by mucus-released larvae is discussed in a broader context.

Background: Natural infection with a species of Angiostrongylus has been reported only once in wildcats from central Italy by Biocca in 1957. The causative species of this infection was identified as Angiostrongylus chabaudi. Following this report, this parasite had never been found in either wild or domestic cats., Findings: The lungs and the pulmonary arteries of an adult female cat (Felis silvestris catus), road-killed in Sardinia, Italy, were macroscopically examined and dissected under a light microscope for the presence of parasites. A slender nematode was detected and its morphometrical features were consistent with those of A. chabaudi. Morphological data were supplemented by sequencing of the partial cytochrome oxidase c subunit 1 (cox1) gene, as well as the internal transcribed spacer 2 (ITS2) of the rDNA. Nucleotide sequences displayed 99% homology with the ITS2 sequence [GenBank KM216825.1] of a specimen of Angiostrongylus sp. recovered recently from the pulmonary artery of a wildcat in Germany and 91% with cox1 sequence [GenBank GU138118.1] of Angiostrongylus vasorum., Conclusion: The results of the present study indicate, for the first time, that A. chabaudi may also infect domestic cats, and thus should be considered in the diagnosis of metastrongyloid species infecting their cardio-pulmonary system.

To investigate larval development of Acanthocheilonema reconditum in the cat flea Ctenocephalides felis, fleas were fed through an artificial feeding system with dog blood containing different concentrations of microfilariae (i.e. low, group L = 250; medium, group M = 500; high, group H = 1500 microfilariae per mL) or no microfilariae (group C). Fleas were sampled at 12 different time-points throughout the study period (D1-D28) and A. reconditum was detected by dissection, PCR and histology. Of 2105 fleas fed with infected dog blood, 891 (38·7%) died during the study before being sampled whilst the remaining (n = 1214) were examined for A. reconditum. Upon dissection, first-stage larvae (L1) were identified after 2 days post infection (D2), second-stage (L2) at D13 and infective third-stage larvae (L3) at D15. Eighteen (30%) of 60 pools of fleas molecularly examined tested positive. Histologically, L2 were detected at D13 in the sub-cuticle region embedded in the back muscle of one female flea. This study provides original data on larval development of A. reconditum in C. felis and reports on the usefulness of the artificial feeding system.

Oslerus rostratus syn. Anafilaroides rostratus (Strongylida: Filaroididae) is a metastrongyloid transmitted by snails, which localizes in peri-bronchial tissues and in the lung parenchyma of wild as well as domestic cats. In Europe, this nematode has been reported only on two occasions, being diagnosed in cats from Majorca Island and in northern Spain. Here, we describe a case of O. rostratus infection in a necropsied 4-year-old cat in Sicily (southern Italy). At the inspection of lungs, slender and greyish nematodes (four females and two males) were found embedded in the peri-bronchial tissues and in the bronchial walls. Parasites were morphological and molecularly identified as O. rostratus, with their 18S sequences being identical among them and showing a high homology (99%) with those available in public databases. At the histology, nematodes were encapsulated in a pseudo-cystic formation surrounded by an interstitial inflammatory process and fibrous tissue. Lung lesions were mainly represented by peri-luminal fibrosis, hyperplasia and hypertrophy of the bronchial mucosa and glands, respectively. This first record of O. rostratus infection from Italy indicates that this parasite should be included in the differential diagnosis of feline of lungworm infection.

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., (Copyright © 2014. Published by Elsevier B.V.)

This study investigates the organizational culture and associated characteristics of the newly established primary care units (PCUs)-collaborative teams of general practitioners (GPs) who provide patients with integrated health care services-in the Emilia-Romagna Region (RER), Italy. A survey instrument covering 6 cultural dimensions was administered to all 301 GPs in 21 PCUs in the Local Health Authority (LHA) of Parma, RER; the response rate was 79.1%. Management style, organizational trust, and collegiality proved to be more important aspects of PCU organizational culture than information sharing, quality, and cohesiveness. Cultural dimension scores were positively associated with certain characteristics of the PCUs including larger PCU size and greater proportion of older GPs. The presence of female GPs in the PCUs had a negative impact on collegiality, organizational trust, and quality. Feedback collected through this assessment will be useful to the RER and LHAs for evaluating and guiding improvements in the PCUs., (© 2013 by the American College of Medical Quality.)

Troglostrongylus brevior is a neglected feline lungworm species, which has been increasingly reported in the Mediterranean area, although scant data are available on the respiratory alterations it causes in cats. Therefore, we describe the gross and histological lesions of a 20-week old kitten that succumbed due to the onset of a fulminant respiratory failure. At necropsy, a catarrhal exudate was observed in the airways, along with nematodes in the trachea and bronchi. The lungs were processed for histological examination and serial pulmonary sections were performed. A total of 14 nematodes were collected, being all morphologically and molecularly identified as T. brevior. Lungworms were histologically localized within the bronchial lumen, surrounded by an eosinophilic infiltrate. The presence of T. brevior in the airways has been histologically documented for the first time and its life-threatening potential is discussed., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: The efficacy of a slow-release insecticidal and repellent collar containing 10% imidacloprid and 4.5% flumethrin (Seresto, Bayer Animal Health) in preventing Leishmania infantum infection was evaluated in a large population of dogs living in a hyper-endemic area of Sicily (Italy)., Methods: A total of 219 dogs, negative for L. infantum were enrolled in a multicentre, controlled study. Dogs were divided into two homogeneous groups, defined as G1 (n = 102) and G2 (n = 117). Before the start of the sand fly season, dogs in G1 were treated with the collar while animals in G2 were left untreated, serving as negative controls. Dogs were serially sampled on day D90, D180, D210 and D300 in order to assess Leishmania infection by IFAT, PCR on skin (D210-D300) and bone marrow (D300) and cytology on bone marrow aspirate (D300)., Results: Three dogs (2.9%) in G1 and 41 (40.2%) in G2 became positive for L. infantum in at least one of the diagnostic tests employed in the study. The number of seropositive dogs in G2 increased in the course of the study from 15 (D90) to 41 (D300), with some of them also positive in other diagnostic tests. Eight (19.6%) of the seropositive dogs in G2 showed an increase in antibody titers ranging from 1:160 to 1:1,280. At the last follow-up, some of dogs in G2 displayed overt clinical signs suggestive of leishmaniosis. The mean incidence density rate at the final follow-up was 4.0% for G1 and 60.7% for G2, leading to a mean efficacy of the collar in protecting dogs at both sites of 93.4%., Conclusions: The slow-release collar tested in this study was shown to be safe and highly effective in preventing L. infantum infection in a large population of dogs. Protection conferred by a single collar (up to eight months) spanned an entire sand fly season in a hyper-endemic area of southern Italy. The regular use of collars, at least during the sand fly season, may represent a reliable and sustainable strategy for the prevention of leishmaniosis in dogs living in or travelling to an endemic area.

The activity of phlebotomine sand flies was monitored in a sub-urban area of Sicily in order to acquire data on seasonality and to elucidate the effect of the night time-intervals, height of traps from ground and lunar phases on the abundance of the capture. The study was conducted in the farm of the University of Messina (Italy). Light traps were placed as in the following: biweekly, from dusk to dawn, and from May to November; for three consecutive nights from 18:00 to 6:00, with the net bag being changed every 2h; for 30 days, at different heights from 18:00 to 6:00. A total of five species (i.e., Phlebotomus perniciosus, Phlebotomus neglectus, Phlebotomus sergenti, Phlebotomus perfiliewi, and Sergentomyia minuta), three of which are proven vectors of Leishmania infantum, were captured. The most abundant species was P. perniciosus (73.3%) followed by S. minuta (23.3%). The highest number of phlebotomine sand flies was collected in August and September with a peak of collection recorded in the evening (i.e., from 20:01 to 22.00). The number of phlebotomine sand flies collected at 50cm above the ground was significantly higher (P=0.041) than that captured at 150cm. Results of this study shed light on the ecology of main phlebotomine species in the Mediterranean area, and on the influence of some factors, such as time and height of traps, on the light trap capture efficiency., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Aelurostrongylus abstrusus (Strongylida, Angiostrongylidae) and Troglostrongylus brevior (Strongylida, Crenosomatidae) are regarded as important lungworm species of domestic felids, with the latter considered an emerging threat in the Mediterranean region. The present study aimed to assess their concurrent development in the mollusc Helix aspersa (Pulmonata, Helicidae). Thirty snails were infested with 100 first-stage larvae (L1) of A. abstrusus and T. brevior, isolated from a naturally infested kitten. Larval development was checked by digesting five specimens at 2, 6 and 11 days post infestation. Larvae retrieved were morphologically described and their identification was confirmed by specific PCR and sequencing. All H. aspersa snails were positive for A. abstrusus and T. brevior, whose larval stages were simultaneously detected at each time point. In addition, snails were exposed to outdoor conditions and examined after overwintering, testing positive up to 120 days post infestation. Data herein presented suggest that A. abstrusus and T. brevior develop in H. aspersa snails and may eventually co-infest cats. Data on the morphology of both parasitic species in H. aspersa provide additional information on their development and identification, to better understand the population dynamics of these lungworms in receptive snails and paratenic hosts.

Background: Cercopithifilaria bainae is a filarioid parasite that infects dogs, being transmitted by Rhipicephalus sanguineus group ticks in many countries of the Mediterranean basin. This study assessed the incidence density rate (IDR) of infection by C. bainae in dogs and the probability of co-infection with other tick-borne pathogens (i.e., Anaplasma platys, Babesia vogeli and Hepatozoon canis), in an area of high endemicity in southern Italy., Methodology/principal Findings: From March 2011 to October 2012, a field study involving 58 young dogs naturally exposed to tick infestation was conducted. Skin and blood samples obtained from each dog six times during an 18-month period were tested for C. bainae by parasite detection within skin snip sediments, with subsequent confirmation through PCR and DNA sequencing. Dogs examined monthly for ticks and A. platys, B. vogeli and H. canis were microscopically and/or molecularly diagnosed and after the first and the second summer seasons, the IDR for positive animal-month at risk was 3.8% and 1.7% in November 2011 and October 2012, respectively. All 58 C. bainae-infected dogs were simultaneously infected with at least one other tick-borne pathogen. After the first summer season (assessment in November 2011), a C. bainae-infected dog had a 33% probability of being infected with H. canis or A. platys, whereas after the second tick season (assessment in October 2012) the probability of co-infection was 78%, 22% and 11% for H. canis, A. platys and B. vogeli, respectively., Conclusions: Our data indicate that tick-infested dogs are at risk of acquiring infection by C. bainae. In addition, the detection of C. bainae microfilariae indicates a prior tick exposure and, should stimulate testing for other tick-borne disease causing pathogens.

In addition to Aelurostrongylus abstrusus (Strongylida: Angiostrongylidae), referred to as the feline lungworm, Troglostrongylus brevior (Strongylida: Crenosomatidae) has recently been identified as an agent of bronco-pulmonary infestations in cats. These two parasites have a similar biology, share ecological niches, potentially co-infesting cats, but are difficult to be differentiated due to the morphological similarities of their first-stage larvae (L1). This paper describes a molecular tool, based on single-step duplex polymerase chain reaction (duplex-PCR) on the ribosomal internal transcribed spacer 2 region (ITS-2) for the simultaneous detection and differentiation of T. brevior and A. abstrusus. L1 of both species were collected from faecal samples, morphologically identified, and single larval specimens isolated. An aliquot of faeces was used as a test sample for a case of mixed natural infestation. The duplex-PCR was performed using species-specific forward primer sets for the ITS-2 region (i.e., A. abstrusus: 220bp; T. brevior: 370bp). The detection limit of the molecular assay was also assessed by serial dilutions of DNA from single larvae of both species (from ≈ 4.0 to 4.0 × 10(-5) μg/μl). The duplex-PCR carried out on individual DNA samples was able to detect as low as 5.2 × 10(-3) μg/μl of DNA for A. abstrusus, 4.9 × 10(-3)μg/μl for T. brevior, and as low as 4.0 × 10(-3) μg/μl for samples containing both species. Species-specific bands of the expected sizes and two bands were simultaneously amplified from the faecal sample containing both species. The phylogenetic analyses of the ITS-2 sequences here examined and those available for other metastrongyloids were concordant in clustering them with those of other Troglostrongylus brevior and A. abstrusus sequences available in GenBank database. This molecular approach proved to be effective and highly sensitive for the simultaneous detection of the two lungworms species and it might be used for molecular epidemiological studies and for monitoring therapeutic protocols., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Dirofilaria repens is one of the most widespread mosquito-borne filarioid responsible for infestations in dogs, usually characterized by nodules and other skin lesions. Additionally, the zoonotic potential of this nematode is of public health concern, given that D. repens has been often associated to human dirofilariosis in the Old World. In the present study, the therapeutic efficacy against D. repens microfilariae of a protocol based on doxycycline hyclate (Ronaxan(®), Merial), 10mg/kg every day for 30 days, and ivermectin (Cardotek 30(®), Merial), 6 μg/kg every 15 days for 6 months, was preliminarily investigated in two naturally infested dogs. Blood samples were collected every 30 days during 6 months of treatment and examined by a Knott's modified test and a duplex real-time PCR. The results here presented indicate that this therapeutic protocol, applied monthly, could be effective for treating microfilariaemia in dogs affected by subcutaneous dirofilariosis., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Troglostrongylus brevior has recently been recognised as a neglected causative agent of broncho-pulmonary infestation in domestic cats. Nonetheless, information on the biology and ecology of this nematode is still scarce. In this study, the survival of T. brevior first-stage larvae (L1) was evaluated in water and in faecal samples at different controlled temperatures in the laboratory (i.e., 4 and 26°C) or in the outdoor environment (mean temperature 14±3.1°C). Vitality of larvae was microscopically assessed every 7days, until their death. Larval survival ranged from 7days in both water and faeces at 26°C up to 142days at 4°C, respectively. Larvae maintained in the environment (14°C) survived from 35 up to 63days in faeces and water, respectively. Data herein presented clearly indicate that the survival of T. brevior L1s is negatively correlated with temperature (P<0.001). According to the results of this study, the infection in mollusc intermediate hosts in the Mediterranean area may easily occur during winter/spring, while during hot and dry seasons a reduction of transmission should be expected., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Recently, dermal microfilariae of a Cercopithifilaria species (Spirurida, Onchocercidae), namely Cercopithifilaria bainae , were detected in dogs from several geographical areas of the Mediterranean basin. Evidence from both laboratory and field studies support the role of the brown dog tick, Rhipicephalus sanguineus sensu lato, as an intermediate host of this nematode. In the present study, we investigated the competence of Ixodes ricinus nymphs as vectors of C. bainae. On November 2012, fully engorged nymphs of I. ricinus (n = 174) and R. sanguineus s.l. (n = 10) were collected from a dog infected by C. bainae. The presence of C. bainae in I. ricinus was assessed by both microscopic dissection of specimens and detection of nematode DNA (PCR), at days 3, 10, 20 and 30 (T1-T4) post-collection; due to the small number of specimens available, R. sanguineus s.l. were examined using the same methods at T4 only. No developing larva of C. bainae was detected in I. ricinus specimens at different time points (T1-T4), even if four of these specimens were PCR-positive at T1. Seven out of ten R. sanguineus s.l. were positive for C. bainae third-stage larvae (L3) at both microscopical and molecular analysis at T4. This study indicates that C. bainae does not develop in I. ricinus nymphs, which preclude the role of this tick as an intermediate host of this parasite. Data presented herein provide new insights into the biology of this filarioid species and will lead to a better understanding of the role of different tick species as vectors of nematodes.

Background: Dog communities living in kennels are at high risk of being infected by ticks and fleas. In spite of the indubitable efficacy of several topical ectoparasiticides registered for the control of ectoparasites in dogs, the short period of action and the price of these products limit their use in dog communities. This paper reports on the efficacy of imidacloprid/flumethrin slow release collars to cure dogs highly infested with ectoparasites and to prevent infestations for 8 months in a refuge with a history of unsuccessful environmental treatments., Methods: A total of 82 dogs were collared with slow release collars containing a formulation of imidacloprid 10% / flumethrin 4.5%. Dogs were followed-up for ectoparasite presence after 2, 7 and 14 days and thereafter biweekly up to 90 days. Furthermore, dogs were examined for ectoparasites whilst replacing collars 8 months after their application., Results: At the time of treatment 79 (96.3%) out of 82 included dogs were heavily infested by ticks and 53 (68.8%) out of the 77 combed dogs were infested by fleas. Tick infested dogs had an estimated mean intensity of 46.9 (± 65.7), while flea infested animals had a load between 20 and 50 fleas. In addition, some of the flea infested dogs (18.9%) were presenting signs of flea allergic dermatitis (FAD). Two days after treatment, 49 (60.5%) and 9 (11.7%) dogs were still infested by live ticks and fleas, respectively. However, the mean intensity of ticks decreased to 3.5 (± 4.3) with a reduction of 92.5%. Except for sporadic cases, no attached ectoparasites were found on dogs from the day 14 visit until the end of the investigation. Cases of FAD resolved without any other treatment within 30 days., Conclusions: Ticks and fleas represent a constant hazard for dog populations. Therefore, in particular settings, such as dog refuges, sustainable and long-term strategies to control ectoparasite infestations are needed. Based on the observed evidence of efficacy, long-term duration and safety, the imidacloprid/flumethrin slow release collars can be regarded as an efficacious and sustainable means for ectoparasite control and for treatment of FAD in high-risk dog communities.

Metastrongyloids of cats are emerging pathogens that may cause fatal broncho-pulmonary disease. Infestation of definitive hosts occurs after ingestion of intermediate or paratenic hosts. Among metastrongyloids of cats, Troglostrongylus brevior and Troglostrongylus subcrenatus (Strongylida: Crenosomatidae) have recently been described as agents of severe broncho-pulmonary disease. Here, we provide, for the first time, observational evidence suggesting the direct transmission of T. brevior from queen cat to suckling kittens. This new knowledge will have a significant impact on current scientific information of this parasite and shed new light into the biology and epidemiology of metastrongyloid nematodes.

Background: Three species of the genus Cercopithifilaria have been morphologically and molecularly characterized in dog populations in southern Europe: Cercopithifilaria grassii (Noè, 1907), Cercopithifilaria sp. sensu Otranto et al., 2011 (reported as Cercopithifilaria sp. I), and Cercopithifilaria sp. II sensu Otranto et al., 2012. The adults of Cercopithifilaria sp. I have remained unknown until the present study., Methods: The material originated from a dog from Sardinia (Italy) diagnosed with dermal microfilariae of Cercopithifilaria sp. I. The holotype and three paratypes of Cercopithifilaria bainae Almeida & Vicente, 1984, described from dogs in Brazil, were studied as comparative material. A cox1 (~689 bp) and 12S (~330 bp) gene fragments were amplified and phylogenetic analysis carried out., Results: The highest numbers of adult nematodes (82%) were collected in the sediment of the subcutaneous tissues of the trunk (n = 37) and forelimbs (n = 36). The morphology of the adult nematodes and microfilariae collected from the dog in Sardinia corresponded to those of C. bainae. All cox1 and 12S gene sequences showed a high homology (99-100%) with sequences from microfilariae of Cercopithifilaria sp., I Conclusions: The morphological and molecular identity of the microfilariae of C. bainae overlap those described previously as Cercopithifilaria sp. sensu Otranto et al., 2011 (=Cercopithifilaria sp. I). Therefore, the present study reports the occurrence of C. bainae in Europe, for the first time after its description and the single record in Brazil. C. bainae appears to be highly diffused in dog populations in southern Europe. The phylogenetic analyses based on cox1 and 12S do not reveal the three species of Cercopithifilaria parasitizing dogs as a monophyletic group, which suggests that they have derived independently by host switching.

Following the recent description of microfilariae of a Cercopithifilaria sp. in a dog from Sicily, Italy, (herein after referred to as Cercopithifilaria sp. I), numerous skin samples were collected from dogs in the Mediterranean region. In addition to Cercopithifilaria sp. I (185·7 ± 7·2 μm long), microfilariae of 2 other species were identified, namely Cercopithifilaria grassii (651·7 ± 23·6 μm long) and a yet undescribed microfilaria, Cercopithifilaria sp. II (264·4 ± 20·2 μm long, with evident lateral alae). The morphological differentiation among the 3 species of dermal microfilariae was confirmed by differences in cytochrome c oxidase subunit 1 and ribosomal 12S sequences examined (mean level of interspecific pairwise distance of 11·4%, and 17·7%, respectively). Phylogenetic analyses were concordant in clustering these with other sequences of Cercopithifilaria spp. to the exclusion of Dirofilaria spp., Onchocerca spp. and Acanthocheilonema spp. Dermal microfilariae collected (n = 132) were morphologically identified as Cercopithifilaria sp. I (n = 108, 81·8%), Cercopithifilaria sp. II (n = 17, 12·9%), whereas only 7 (5·3%) were identified as C. grassii. Mixed infestations were detected in all sites examined. The great diversity of these neglected filarioids in dogs is of biological interest, considering the complex interactions occurring among hosts, ticks and Cercopithifilaria spp. in different environments.

The aim of this study was to investigate the anatomical distribution of microfilariae of a recently described Cercopithifilaria sp. and the relationship with the preferred attachment sites of its vector, the brown dog tick Rhipicephalus sanguineus. Skin samples from 20 dogs were collected from eight anatomical sites and soaked twice in saline solution at 37°C. All samples were also molecularly processed for the specific amplification of partial cytochrome c oxidase 1 gene. Microfilariae were unevenly distributed on the body, with higher frequencies on interscapular region (n=13; 68.4%) and on the head (n=9; 47.4%). Larval abundance in dogs was positively correlated with the number of positive anatomical sites with a higher percentage of skin samples scoring positive at the microscopic examination of the first sediment (n=47; 30.9%) than of the second (n=8; 5.3%). Histological examination revealed that microfilariae were scattered in the dermis, in association with inflammatory cells. Molecular analysis of skin samples showed a lower frequency of positive sites (n=37; 24.3%) when compared to the microscopic examination (n=53; 34.9%) with five haplotypes sequenced. The results here presented suggested a close co-evolution of this filarioid with its vector., (Copyright © 2012 Elsevier B.V. All rights reserved.)

In spite of its wide distribution among dogs and the evidence of its implication as a zoonotic agent, scant information is available on the biology of Acanthocheilonema reconditum (Spirurida, Onchocercidae). In this study, blood samples from 152 Sicilian dogs were examined for A. reconditum microfilariae at the beginning of the study and 1 year later. The periodicity of microfilaraemia was investigated by bleeding 2 highly microfilaraemic dogs twice a day for 10 days and, later on, every 2 weeks for 1 year and a third animal every 3 h for 96 h. Fleas and ticks infesting dogs were collected and dissected for the detection of A. reconditum larvae. The prevalence of infestation was 11·2% (17/152) and 13·3% (16/120) at the beginning and at the end of the study, with a 1 year cumulative incidence of 5·9%. Although dogs bled twice a day showed a higher number of microfilariae in most of the morning samples, the absence of any circadian rhythm was suggested by data of the third experiment conducted by bleeding a dog every 3 h for 4 days. A. reconditum developing forms were detected in 5·1% (4/78) of dissected fleas, but not in any of the 272 ticks. The study provides new insights into the biology and ecology of this dog filarioid in its definitive and intermediate hosts.

This study investigates sequence variation in mitochondrial cytochrome c oxidase subunit 1 gene within Cercopithifilaria sp. recorded recently in Italy. Fourteen sequence types (haplotypes) were characterized for 163 (7.7%) amplicons from 2111 Genomic DNA samples prepared from skin samples from dogs and from Rhipicephalus sanguineus (ticks) from different geographical areas of the Mediterranean basin (i.e., Italy, Spain and Greece). The most prevalent sequence types represented haplotypes I (70.5%) and X (16.0%), followed by haplotype VIII (4.9%) and other 11 haplotypes (8.6%). Three haplotypes (II, V and VI) were found exclusively in ticks. The overall intraspecific nucleotide variation among pcox1 haplotypes ranged from 0.4 to 3.5% (mean = 1.6%), whereas a mean interspecific difference of 9.5% was detected as compared with other onchocercids. Phylogenetic analysis of the nucleotide sequence data showed a clustering of Cercopithifilaria sp. with the other Cercopithifilaria species (with strong statistical support) to the exclusion of other onchocercids. The number of haplotypes identified here might be explained by complex ecology and transmission patterns as well as the high mutation rate of mitochondrial DNA and/or inbreeding associated with hosts and their vectors., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The present study reports the applicability of a multiplex PCR for the simultaneous detection and differentiation of common filarioids infecting dogs, i.e., Dirofilaria immitis, Dirofilaria repens, Acanthocheilonema reconditum and Cercopithifilaria sp. Amplicons of different sizes (i.e., 170 bp, 480 bp, 590 bp and 300 bp, respectively) of regions within the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene were amplified on a single-step multiplex PCR using a mix of species-specific forward primers coupled with a single reverse primer. Experiments were carried out by amplifying genomic DNA extracted from blood or skin samples test-positive for microfilariae (mff). The number of mff present in each blood sample was quantified (from 800 to 25,000 mff/ml for A. reconditum and D. repens, respectively) and mixed blood samples were tested for the simultaneous detection of DNA from these mff. Specific amplicons for blood-circulating mff of A. reconditum, D. immitis and D. repens and for those whose adults are localized in skin (i.e., A. reconditum and Cercopithifilaria sp.) were simultaneously detected on agarose gel up to a dilution of 250 mff/ml for D. repens. The specific identity of the amplicons was confirmed by sequencing. The multiplex PCR assay reported herein represents a new tool for the molecular detection and differentiation of canine filarioids in blood and skin samples., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Dermal microfilariae found in a dog from Sicily, Italy, were characterized morphologically and genetically and differentiated from those of all the other blood microfilariae commonly found in dogs. In particular, the microfilariae were short (mean length of 186.7 μm), presented a body flattened dorso-ventrally and a rounded head, bearing a tiny cephalic hook. The genetic identity of microfilariae herein studied was also assessed by molecular amplification, sequencing and analyzing of multiple ribosomal ITS-2 and mitochondrial (cox1 and 12S) target genes. Both morphologic and genetic characterization as well as the molecular phylogenetic history inferred using sequences of a barcoding dataset were concordant in supporting the identification of Cercopithifilaria at the genus level. Surprisingly, microfilariae here examined were well distinct from Cercopithifilaria grassii (Noè, 1907), from northern Italy, and resembled those of a species described in Brazil, Cercopithifilaria bainae Almeida & Vicente, 1984. This paper provides evidence for the existence of a Cercopithifilaria species infesting a dog from Sicily and also presents a PCR protocol on skin samples as a tool for further epidemiological studies, which could provide evidence on the aetiology and the natural history of this filarial species., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A trial to assess the efficacy of eprinomectin (EPM) against the lungworm Dictyocaulus arnfieldi was carried out on 15, naturally-infected donkeys. Ten animals were treated with a 'pour-on' EPM preparation (at a dose of 0.5 mg/kg bodyweight), and five animals acted as controls. Faecal larval counts were carried out two days before treatment, on day of treatment and 7, 14, 21 and 28 days post-treatment with the anthelmintic. EPM was 100% effective in eliminating faecal larvae from day 7, until the end of study at day 28. No adverse drug-reactions or side-effects were observed in any of the treated donkeys., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Wohlfahrtia magnifica (Schiner, 1862; Diptera: Sarcophagidae), known also as flesh fly or screwworm, is a fly distributed especially in south Europe, Middle East, North Africa and China. Maggots of W. magnifica are responsible for traumatic myiasis in warm-blooded vertebrate animals and humans. In Italy data on wohlfahrtiosis in animals and humans are scant. This paper reports three cases of genital myiasis by W. magnifica in Sicily that occurred in a goat, a ram and a dog, respectively. Maggots were found in the vulva of a goat, in a wound localised in the scrotum of a ram and in the prepuce of a dog. Although the disease was not recorded before in the region, data collected from a questionnaire specifically designed for the study suggest that the disease is endemic in the area for more than 20 years. Larger epidemiological studies in the region are needed to get more data on disease prevalence and pattern of infestation as well as more information and education is needed for both farmers and veterinarians on wohlfahrtiosis and its strategies of control and treatment.

Background: Thelazia callipaeda (Spirurida, Thelaziidae), eyeworms, are known as the causative agents of thelaziosis, initially described in Asia and, later on, over the last decade, also in some European countries (e.g., Italy, France, Germany and Switzerland). In June 2010, the first case of canine thelaziosis was observed in central western Spain (La Vera region, Cáceres) and subsequent epidemiological investigation is reported in the present study., Results: This study describes the first autochthonous cases of infection by T. callipaeda in dogs from central western Spain where the first case of eyeworm infection was reported.A total of 456 dogs was examined in this geographical area. Thelazia callipaeda eyeworms were observed in 182 (39.9%) animals, of which 28 showed apparent clinical signs (i.e., conjunctivitis, oedema, epiphora and petechiae). A total of 762 adult nematodes (214 males, 548 females; mean infection rate of 4.18; SD 4.74) were collected with cotton swabs or by flushing of the conjunctival sac of infected animals using physiological saline solution. Nematodes were identified as T. callipaeda according to the morphological keys and molecular analysis of sequences of a portion of the mitochondrial cytochrome c oxidase subunit 1 (cox 1) gene. The sequences were identical to those representing T. callipaeda haplotype 1, previously reported in Europe., Conclusions: The high infection rate of canine thelaziosis herein reported suggests that practitioners should include this eye infection amongst differential diagnoses of ocular diseases in dogs from this area of Spain or those moving across this area of Spain. Based on the high infection prevalence recorded, the potential public health risk to humans from this region is also discussed.

Dirofilaria repens, which is usually found in canine subcutaneous tissues, is the main causative agent of human dirofilariasis in the Old Word. However, a relationship between animal and human cases of dirofilariasis caused by D. repens in a given area has never been demonstrated. The uneven distribution of D. repens in provinces in Sicily, Italy represented the foundation for this study. We report a human case of ocular infection with D. repens from Trapani Province, where canine dirofilariasis is endemic. The nematode was morphologically and molecularly identified and surgical removal of the parasite was documented. The relationship between the prevalence of D. repens in dogs and the occurrence of human cases of ocular dirofilariasis is discussed on the basis of a review of the historical literature.