Experimental Study of the Mechanical Transmission of Rabbit Hemorrhagic Disease Virus (RHDV2/b) by Aedes albopictus (Diptera: Culicidae) and Phlebotomus papatasi (Diptera: Psychodidae).

Rabbit hemorrhagic disease (RHD) is caused by a lagovirus mainly affecting European rabbits (Oryctolagus cuniculus), although other European and North American lagomorph species are also susceptible to fatal infection by the new viral variant RHDV2/b. In the present work, direct mechanical transmission of the rabbit hemorrhagic disease virus (RHDV2/b variant) by the hematophagous Diptera Aedes albopictus (Skuse) (Diptera: Culicidae) and the sand fly Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) was tested. For each species, six and three laboratory rabbits were exposed to bites of dipterous females partially fed on RHDV2/b viral suspension 2 h and 24 h prior to exposure, respectively. The rabbits were then monitored for clinical changes and mortality for 35 d, and seroconversion was assessed by indirect ELISA. No rabbit died or showed clinical signs of disease, and seroconversion was recorded in two rabbits challenged with P. papatasi females fed the viral suspension 2 h prior to exposure. The number of RHDV2/b RNA copies/female was higher in Ae. albopictus than in P. papatasi but the decrease over time of RNA load in Ae. albopictus was greater than that in P. papatasi. The results of this study suggest the inability of Ae. albopictus to serve as a direct mechanical vector of RHDV2/b, but sand flies could play a role in the local transmission of RHD.

Epidemiological patterns of bovine besnoitiosis in an endemic beef cattle herd reared under extensive conditions.

Bovine besnoitiosis is a parasitic disease caused by the protozoan Besnoitia besnoiti. Described many decades ago, recent epidemiological studies reveal its important spread within Europe in the last years. To date, many epidemiological aspects related to life cycle, routes of transmission, incidence rates and associated risk factors are lacking; hence, the establishment of appropriate disease control programmes poses an important challenge. Thus, the aim of the present study was to determine the epidemiological pattern of the disease in an endemic herd reared under extensive conditions (Spanish Pyrenees) by identifying main factors associated with infection and clinical disease dynamics. The study population consisted of 276 Brown Swiss and Pirenaica adult animals and 145 calves born and weaned at the farm during the study. Three sampling time frames were used: January 2010, September 2010 and February 2011, which allowed us to differentiate two periods designated as mountain and valley periods. The data related to animals (breed, sex and age) and herd management (animal grouping and time in housing) were recorded. The data collection methodology was mainly based on clinical examinations and defining the serological status against bovine besnoitiosis by the immunofluorescent antibody testing of blood samples. The total prevalence among adult animals was 38.34% (CI95%: 34.53-42.07), with 18.54% of seropositive animals showing clinical signs. In regard to the cumulative incidence, 34.57% of new infections were detected during the mountain period, in contrast to the 24.59% observed in the valley period. The incidence density was 0.058 and 0.061 new infections per animal-month for the mountain and valley periods, respectively. According to the seroepidemiological study, the seroconversion probability of B. besnoiti infection was directly associated with the number of seropositive cows with whom an animal had been stabled as well as the housing period duration, supporting horizontal transmission by close contact as one of the most important methods of disease spread. In addition, the risk of developing the clinical course increased with age, and the presence of clinical signs was related to higher antibody responses. Among calves (from 3.1 to 7.1 months old) sampled once at weaning, the total seroprevalence was 15.17% (CI95%: 9.36-21.04), and the chronic stage was observed in three animals, supporting the ability of B. besnoiti to infect and even cause disease in animals less than 6 months old. Finally, the risk of calf seroconversion was positively related to the serological status of the cows, suggesting postnatal transmission between dams and offspring by contact during the suckling period.

Variability of the egg hatch assay to survey benzimidazole resistance in nematodes of small ruminants under field conditions.

The egg hatch assay (EHA) is one of the main in vitro methods for detection of benzimidazole resistance in nematode parasites of small ruminants. However, although the EHA has been standardised at the laboratory level, the diagnostic performance of this method has not been fully characterised for field screenings. In the present work, monthly variation of benzimidazole resistance estimated by EHA was surveyed over two years in three sheep flocks and in one goat and an additional sheep flock sharing the same pastures. Resistance was measured by calculating both the effective dose of thiabendazole (TBZ) that inhibited hatching of ≥50% of parasite eggs (ED50) and the proportion (Pdd) of eggs hatching at a discriminating dose of 0.1 µg/ml TBZ. Pdd exhibited higher variability than ED50, in agreement with the higher sensitivity of Pdd to changes in resistance levels. Both resistance parameters, however, were highly correlated, and their variation was similarly related to the same factors. Resistance levels differed among sheep flocks, and the resistance level of the goat flock was higher than that measured for the sheep flock sharing the same pasture. Moreover, monthly variation of resistance in goats did not mirror that recorded in sheep. Resistance levels varied seasonally, with the highest values recorded in the spring, and they were inversely related to the number of days that samples were stored under anaerobic conditions. In addition, they were directly associated with the relative abundance of Teladorsagia spp. but inversely related to the relative abundance of Haemonchus spp. After controlling for the effects of these identified factors for variation, inter-monthly sampling variation due to unknown factors was the main source of variability, accounting for more than 60-70% of variance for both resistance parameters and yielding absolute estimation errors higher than 0.06 for ED50 or 0.2 for Pdd when resistance was estimated from a single sampling. Optimum sample size, estimated from variance components, suggested that at least 4-5 samplings would be needed to halve this absolute error, whereas additional samplings would slightly increase precision but at the cost of substantially increasing sampling effort. More research is needed to identify the main factors involved in this inter-sampling variation to standardise the implementation of EHA under field conditions.

Sensitivity of Culicoides obsoletus (Meigen) (Diptera: Ceratopogonidae) to deltamethrin determined by an adapted WHO standard susceptibility test.

Bluetongue is a disease of major economic concern in Europe. Its causative agent, bluetongue virus (BTV), is transmitted by several Culicoides species (mainly Culicoides imicola and Culicoides obsoletus in Europe). The application of insecticides on animals may reduce transmission of BTV, however, no formulation is currently licensed specifically against Culicoides midges. The present study assesses the susceptibility of C. obsoletus to deltamethrin using an adapted World Health Organization (WHO) susceptibility test. Midges were exposed to different dosages of deltamethrin for 1 h, and mortality after 1 h and 24 h was recorded. Results indicated that deltamethrin is highly toxic to C. obsoletus since a dose of 1·33×10(-4)% was enough to kill 50% of the population (LD50) in 24 h. The deltamethrin concentration needed to kill 90% of the population (LD90) was 5·55×10(-4)%. The results obtained in the present work could help to create a system that can be used to assess insecticide resistance and susceptibility of Culicoides biting midges.

Abundance modelling of invasive and indigenous Culicoides species in Spain.

In this paper we present a novel methodology applied in Spain to model spatial abundance patterns of potential vectors of disease at a medium spatial resolution of 5 x 5 km using a countrywide database with abundance data for five Culicoides species, random regression Forest modelling and a spatial dataset of ground measured and remotely sensed eco-climatic and environmental predictor variables. First the probability of occurrence was computed. In a second step a direct regression between the probability of occurrence and trap abundance was established to verify the linearity of the relationship. Finally the probability of occurrence was used in combination with the set of predictor variables to model abundance. In each case the variable importance of the predictors was used to biologically interpret results and to compare both model outputs, and model performance was assessed using four different accuracy measures. Results are shown for C. imicola, C. newsteadii, C. pulicaris group, C. punctatus and C. obsoletus group. In each case the probability of occurrence is a good predictor of abundance at the used spatial resolution of 5 x 5 km. In addition, the C. imicola and C. obsoletus group are highly driven by summer rainfall. The spatial pattern is inverse between the two species, indicating that the lower and upper thresholds are different. C. pulicaris group is mainly driven by temperature. The patterns for C. newsteadii and C. punctatus are less clear. It is concluded that the proposed methodology can be used as an input to transmission-infection-recovery (TIR) models and R0 models. The methodology will become available to the general public as part of the VECMAP™ software.

Survey of flea infestation in cats in Spain.

Fleas are a common cause of feline skin disorders as well as vectors of zoonotic diseases. This study evaluated the flea species infesting domestic cats in Spain and assessed factors influencing their distribution. Fleas from 217 cats from 57 localities in Spain were identified and associations between abundance, and host-dependent, host habitat and environmental factors were examined. Variations in infracommunity and component community structure were also explored. Three species were present, of which Ctenocephalides felis (Bouché) (Siphonaptera: Pulicidae) was the most abundant (98.4%), followed by Ctenocephalides canis (Curtis) (1.1%) and Pulex irritans (L.) (Siphonaptera: Pulicidae) (0.5%). Overall abundance and abundances of both C. felis and C. canis were higher on farms than in apartments, but overall flea abundance and abundances of both C. felis and C. canis were lower in rural than urban environments. Overall abundance and C. felis abundance were lower during the warmest months, and mean annual rainfall was positively correlated with overall, C. felis and C. canis abundances. No relationship between the number of species per cat and any host, habitat or physiographical variable was found. Species richness was not correlated with mean annual temperature or rainfall. Flea abundance was mainly associated with host habitat and environmental factors.

Collection of Culicoides spp. with four light trap models during different seasons in the Balearic Islands.

Bluetongue (BT) is a viral disease that affects ruminants, being especially pathogenic in certain breeds of sheep. Its viral agent (bluetongue virus; BTV) is transmitted by several species of Culicoides biting midges (Diptera: Ceratopogonidae). Different models of suction light traps are being used in a number of countries for the collection of BTV vector species. To determine the relative effectiveness of different light traps under field conditions, four traps (Onderstepoort, Mini-CDC, Rieb and Pirbright) were compared. These traps were rotated between four sites on a cattle farm in Mallorca (Balearic Islands, Spain) for several non-consecutive nights. Results showed remarkable disparities in the efficacy of the traps for the collection of Culicoides midges. The highest number of midges collected in the Onderstepoort trap (x¯±SD=62±94.2) was not significantly different from that collected in the Mini-CDC (x¯±SD=58±139.2). The Rieb trap collected the lowest number of midges (x¯±SD=3±4.0). Significantly higher mean numbers of midges were collected in the Onderstepoort than in either the Pirbright (P=0.002) or Rieb traps (P=0.008). There were also differences in the Culicoides species composition as determine with the various traps. These results indicate that the Onderstepoort or Mini-CDC traps will be more effective than either the Rieb or Pirbright traps for the collection of large numbers of Culicoides midges.

Host feeding patterns of Culicoides species (Diptera: Ceratopogonidae) within the Picos de Europa National Park in northern Spain.

Blood meal identification can provide information about the natural host-feeding patterns or preferences of Culicoides species. Such information could indirectly provide data indicating which reservoirs are significant in associated vector-borne diseases. We positively identified the host species through DNA sequencing of the cytochrome b gene in 144 of the 170 (84.7%) blood meal specimens tested. In the remaining samples, identification of the blood-meal source was unsuccessful, possibly due to the post-ingestion time prior to sampling or the availability of the species-specific cytochrome b gene sequences in the database. The majority of identified blood meals were derived from mammalian blood (95.8%), and only six contained chicken blood. We identified five species as mammalian hosts for Culicoides spp.: sheep (87.7%), human (6.5%), cattle (3.7%) and Savi's Pine Vole (Micrototus savii) (2.1%). The results suggested that large mammals, specifically ruminants, were most frequently fed upon by biting midges (Culicoides spp.), but evidence of opportunistic feeding behaviour was also found. Host feeding behaviour of Culicoides species may also be influenced by the relative abundance of a particular host species in the area being studied. In this sense, Savi's Pine Vole, a wild species, was found to be a locally relevant host and a putative reservoir for viruses transmitted by species of biting midges belonging to the Culicoides genus. Finally, feeding on multiple potential host species was observed. One midge acquired blood meals from human and chicken hosts, while four other midges fed on two different sheep.

Management and environmental factors related to benzimidazole resistance in sheep nematodes in Northeast Spain.

A survey to determine the level of parasite resistance to benzimidazoles (BZ) under field conditions was performed on 107 commercial sheep farms located in the Aragon region of northeast Spain. Resistance was measured using the discriminant dose, a simplified form of the in vitro egg hatch assay (EHA). Taking into account the spatial structure of the data, a multivariate approach was applied to management and environmental variables as well as to their relationships with BZ resistance levels compiled from each flock. Results estimated that 11% of flocks had resistant parasite populations, although we suspected the presence of BZ-resistant parasite strains in 98% of the sample. Resistance levels were more similar among the nearest flocks, suggesting a contagious spatial distribution of resistance (i.e., resistance at neighbouring farms was not independent from one another). Management variables such as frequency of deworming, grazing in private pastures and underdosing were positively related to resistance levels, whereas only the use of BZ was negatively related to resistance levels, likely because BZ were replaced by other anthelmintics in flocks where reduced BZ efficacy was suspected. In addition to climatic conditions and seasonality, land use was an environmental variable associated with observed BZ resistance levels. Generally, resistance was highest in cooler and wetter areas but was lower in flocks sampled during January-March compared to flocks sampled in April-June or October-December. Variation partitioning procedures showed that the variation of resistance explained by the effect of environmental variables was higher than management variables. The effects of both variable groups, however, highly overlapped with the spatial structure of resistant levels, which suggests that a considerable amount of the effects attributable to both variable groups may be actually due to the spatial distribution of resistance. The resistance variation explained by the spatial component suggested that other uncontrolled factors acting at short spatial scale (e.g., common management and environmental variables; the importation of resistant strains and their posterior spread across neighbouring flocks; the selection history of the worms carried out by historical management events previous to this survey; and genetic, physiological or both types of parasite population variation) could yield this contagious spatial structure of BZ resistance. Although further research is needed, both seasonal variation and the dependence of resistance levels among neighbouring flocks should be taken into account in the design of future research or observational resistance programmes to minimise spatial and temporal pseudo-replication. Thus, research would avoid biased estimations of resistance prevalence or of its relationship with putative factors.

Widespread exposure to Sarcoptes scabiei in wild European rabbits (Oryctolagus cuniculus) in Spain.

Sarcoptic mange was recently described in the wild European rabbit (Oryctolagus cuniculus) in north-eastern Mediterranean Spain, the first such infection reported in this species anywhere in the world. This finding has created concern in conservationists and game managers given that an outbreak of mange after a translocation would have catastrophic consequences for naïve rabbit populations in other parts of Spain. A retrospective serosurvey using an 'in house' ELISA test based on the use of a recombinant antigen aimed at determining the rates of contact with Sarcoptes scabiei was carried out on sera from 966 rabbits collected between 1993 and 2010 in Spain. Antibodies were found in 13% of wild rabbits in 60% of the 53 areas surveyed, as well as in 16 of the 17 Spanish provinces and islands studied. Seropositive rabbits were found amongst the oldest samples analyzed and in all studied years. Antibodies were also detected in 36% of rabbits from the protected island of Dragonera, where rabbits have probably not been released since the 1970s. On Mallorca, where 89 rabbits were inspected for both lesions and antibodies, the prevalence of lesions (5.6%) was much lower than the seroprevalence (22.5%), indicating that rabbits often survive infection or that ELISA detects infected rabbits before they develop visible lesions. Seroprevalence was higher in areas with medium levels of rabbit abundance, no restocking and high rainfall. The results show that mange is widespread in rabbits and that the mite is not a recent introduction. Thus, sarcoptic mange could be considered as an enzootic disease in the wild rabbit and so prophylactic measures implemented during rabbit translocations are to be encouraged to avoid local outbreaks in naïve populations.

Recovery rates of bluetongue virus serotypes 1, 2, 4 and 8 Spanish strains from orally infected Culicoides imicola in South Africa.

Bluetongue (BT) is an infectious disease of ruminants that has spread northwards in Europe during the last decade. The aetiological agent of the disease is an arbovirus [bluetongue virus (BTV)] that belongs to the genus Orbivirus (family Reoviridae). The virus is transmitted by certain species of biting midge within the genus Culicoides (Diptera: Ceratopogonidae). Information on the vector status of the Culicoides species in a specific area will be essential to predict the risk for BTV incursion. Field-collected Culicoides (Avaritia) imicola Kieffer from South Africa were fed on blood containing several Spanish isolates of BTV. Despite the high virus concentrations in the bloodmeal (5.1-6.4 log(10) TCID(50) /mL of blood), virus was recovered from <1% of midges assayed after incubation. Virus concentrations >2.5 log(10) TCID(50) /midge in individual infected C. imicola suggest virus replication with possible risk for transmission to susceptible vertebrate hosts in the field for at least two of the serotypes assayed (BTV-1 and BTV-2). A third serotype (BTV-4) was very close to the estimated threshold for transmission. The relatively low to near refractory status of C. imicola compared with other vector species such as Culicoides bolitinos supports previous results, indicating that Culicoides species other than C. imicola may play a more important role in the epidemiology of BTV.

Absence reduction in entomological surveillance data to improve niche-based distribution models for Culicoides imicola.

Data-driven models for the prediction of bluetongue vector distributions are valuable tools for the identification of areas at risk for bluetongue outbreaks. Various models have been developed during the last decade, and the majority of them use linear discriminant analysis or logistic regression to infer vector-environment relationships. This study presents a performance assessment of two established models compared to a distribution model based on a promising ensemble learning technique called Random Forests. Additionally, the impact of false absences, i.e. data records of suitable vector habitat that are, for various reasons, incorrectly labelled as absent, on the model outcome was assessed using alternative calibration-validation schemes. Three reduction methods were applied to reduce the number of false absences in the calibration data, without loss of information on the environmental gradient of suitable vector habitat: random reduction and stratified reduction based on the distance between absence and presence records in geographical (Euclidean distance) or environmental space (Mahalanobis distance). The results indicated that the predicted vector distribution by the Random Forest model was significantly more accurate than the vector distributions predicted by the two established models (McNemar test, p<0.01) when the calibration data were not reduced with respect to false absences. The performance of the established models, however, increased considerably by application of stratified false absence reductions. Model validation revealed no significant difference between the performance of the three distinct Culicoides imicola distribution models for the majority of alternative stratified reduction schemes. The main conclusion of this study is that the application of Random Forests, or linear discriminant analysis and logistic regression on the condition that calibration data were first reduced on geographical or environmental information, potentially lead toward better vector distribution models.

Protection of livestock against bluetongue virus vector Culicoides imicola using insecticide-treated netting in open areas.

The protection of livestock against Culicoides species (Diptera: Ceratopogonidae) using physical barriers or chemically treated barriers is difficult owing to the small size of these biting midges and animal welfare concerns associated with the reduction of air flow. Culicoides imicola Kieffer is the main bluetongue virus vector in the Mediterranean basin, including the southern Iberian peninsula, where livestock is mainly housed in open pens or sheds which offer no physical protection against C. imicola. In this study we assessed the efficacy of surrounding yearling ewe pens with a canvas barrier or a cypermethrin-treated canvas barrier in reducing the entry of Culicoides spp. and C. imicola. Analyses were based on comparisons of Culicoides catches in traps in pens with and without barriers, and in traps located outside pens. Although there was no clear reduction in the abundance of Culicoides other than C. imicola in pens with either barrier, the C. imicola presence was markedly reduced by the insecticide-treated barrier compared with the untreated barrier; the latter did not reduce the abundance of this species in pens. Estimates of the protection conferred against C. imicola by the treated barrier differed depending on whether catch comparisons were based on outside traps or on traps located inside no-barrier pens. The results suggest that the use of insecticide-treated barriers may reduce contact between livestock and C. imicola in open areas or sheds. More research is necessary to assess the degree of protection as a function of barrier height, C. imicola abundance, and the size of the area to be protected.

Effect of feeding bioactive forages on infection and subsequent development of Haemonchus contortus in lamb faeces.

Contrasting dried herbage diets were offered to lambs to evaluate their effects on Haemonchus contortus infection and on subsequent development of larvae in faeces. Artificially infected lambs (n=24, 4 months old) were allocated to one of four treatment groups (n=6) and fed on hay of lucerne (Medicago sativa; Lu), sulla (Hedysarum coronarium; Su), sainfoin (Onobrychis viciifolia; Sa) or wormwood (Artemisia absinthium; Wo) included as 20% in ground lucerne pellets. While voluntary intake was similar, lamb growth rates were notably affected by diet. Additionally, necropsies carried out 30 days post-infection showed that worm burden was reduced by 8 (Su), 13 (Sa) and 49% (Wo) with respect to that found in lambs fed on lucerne, though only the difference with wormwood was significant. Faecal egg excretion expressed on a dry matter basis was also reduced by 22 (Su), 54 (Sa) and 73% (Wo), but differences were significant only for wormwood and sainfoin, which was associated with a decrease in female fecundity. Sulla and sainfoin diets significantly reduced egg hatching rates but increased the percentage of L(3) in the faeces of the host animal, particularly in the case of sulla hay compared to other forages. Consequently, the potential risk of infection derived from faecal excretion could be notably altered. Further work is needed to validate these findings and their implications in an applied farming system.

Variations in the mitochondrial cytochrome c oxidase subunit I gene indicate northward expanding populations of Culicoides imicola in Spain.

Culicoides imicola is the main vector for bluetongue (BT) and African horse sickness (AHS) viruses in the Mediterranean basin and in southern Europe. In this study, we analysed partial mitochondrial cytochrome c oxidase subunit I (COI) gene to characterize and confirm population expansion of Culicoides imicola across Spain. The data were analysed at two hierarchical levels to test the relationship between C. imicola haplotypes in Spain (n = 215 from 58 different locations) and worldwide (n = 277). We found nineteen different haplotypes within the Spanish population, including 11 new haplotypes. No matrilineal subdivision was found within the Spanish population, while western and eastern Mediterranean C. imicola populations were very structured. These findings were further supported by median networks and mismatch haplotype distributions. Median networks demonstrated that the haplotypes we observed in the western Mediterranean region were closely related with one another, creating a clear star-like phylogeny separated only by a single mutation from eastern haplotypes. The two, genetically distinct, sources of C. imicola in the Mediterranean basin, thus, were confirmed. This type of star-like population structure centred around the most frequent haplotype is best explained by rapid expansion. Furthermore, the proposed northern expansion was also supported by the statistically negative Tajima's D and Fu's Fs values, as well as predicted mismatch distributions of sudden and spatially expanding populations. Our results thus indicated that C. imicola population expansion was a rapid and recent phenomenon.

Entry of bluetongue vector Culicoides imicola into livestock premises in Spain.

Culicoides imicola Kieffer is considered to be the main vector of bluetongue disease (BT) and African horse sickness (AHS) in the Mediterranean basin. It has been assumed that this midge species is exophilic and, consequently, that stabling of livestock should provide effective protection against these diseases. This study presents the results of sampling surveys for C. imicola carried out both inside and outside stables on three farms in mainland Spain. The number of C. imicola captured varied as a function of the populations sampled and trap location (inside vs. outside). The daily mean number captured inside during the sampling of each farm population was directly correlated with the daily mean number captured outside, but daily correlation of captures was not observed. By contrast with previous studies, the mean catch of C. imicola inside was consistently higher than that outside. No clear effect of stable characteristics on the degree of entry was detected. In addition, proportions of males and age-graded female groups varied among populations and with trap location. Proportionately more males and fewer engorged females were captured outside than inside, although the proportions varied among stables. These results contrast with those of previous studies, and with the assumed pronounced exophilic behaviour of C. imicola, and raise important questions about the vector activity of this species in the study area and its implications for the epidemiology of BT and/or AHS.

Ecological correlates of bluetongue virus in Spain: predicted spatial occurrence and its relationship with the observed abundance of the potential Culicoides spp. vector.

Using data from bluetongue (BT) outbreaks caused by viral serotype 4 (BTV-4) in Spain during 2004-2005, a predictive model for BTV-4 occurrence in peninsular Spain was developed. An autologistic regression model was employed to estimate the relationships between BTV-4 presence and bioclimatic-related and host-availability-related variables. In addition, the observed abundances of the main potential Culicoides vectors during 2004-2005, namely Culicoides imicola, Culicoides obsoletus group, and species of the Culicoides pulicaris group, were compared between BTV-4 presence/absence areas predicted by the model. BTV-4 occurrence was mainly explained by bioclimatic variables, although a consideration of host-availability variables led to improved fit of the model. The area of BTV-4 presence predicted by the model largely resembled the core distribution area of C. imicola, and this species was the most abundant Culicoides spp. in predicted BTV-4 presence areas. The results suggest that the spatial expansion of BTV-4 took place only as far as those areas in which C. imicola populations efficiently transmitted the virus.

Modelling the distributions and spatial coincidence of bluetongue vectors Culicoides imicola and the Culicoides obsoletus group throughout the Iberian peninsula.

Data obtained by a Spanish national surveillance programme in 2005 were used to develop climatic models for predictions of the distribution of the bluetongue virus (BTV) vectors Culicoides imicola Kieffer (Diptera: Ceratopogonidae) and the Culicoides obsoletus group Meigen throughout the Iberian peninsula. Models were generated using logistic regression to predict the probability of species occurrence at an 8-km spatial resolution. Predictor variables included the annual mean values and seasonalities of a remotely sensed normalized difference vegetation index (NDVI), a sun index, interpolated precipitation and temperature. Using an information-theoretic paradigm based on Akaike's criterion, a set of best models accounting for 95% of model selection certainty were selected and used to generate an average predictive model for each vector. The predictive performances (i.e. the discrimination capacity and calibration) of the average models were evaluated by both internal and external validation. External validation was achieved by comparing average model predictions with surveillance programme data obtained in 2004 and 2006. The discriminatory capacity of both models was found to be reasonably high. The estimated areas under the receiver operating characteristic (ROC) curve (AUC) were 0.78 and 0.70 for the C. imicola and C. obsoletus group models, respectively, in external validation, and 0.81 and 0.75, respectively, in internal validation. The predictions of both models were in close agreement with the observed distribution patterns of both vectors. Both models, however, showed a systematic bias in their predicted probability of occurrence: observed occurrence was systematically overestimated for C. imicola and underestimated for the C. obsoletus group. Average models were used to determine the areas of spatial coincidence of the two vectors. Although their spatial distributions were highly complementary, areas of spatial coincidence were identified, mainly in Portugal and in the southwest of peninsular Spain. In a hypothetical scenario in which both Culicoides members had similar vectorial capacity for a BTV strain, these areas should be considered of special epidemiological concern because any epizootic event could be intensified by consecutive vector activity developed for both species during the year; consequently, the probability of BTV spreading to remaining areas occupied by both vectors might also be higher.

Fleas parasitizing domestic dogs in Spain.

In addition to their importance to veterinary clinical practice as ectoparasites, fleas of domestic dogs are of special concern because they can be vectors of disease, including zoonoses. Flea assemblages parasitizing domestic dogs usually comprise several flea species whose distribution is determined by factors acting at several scales. Knowledge of these factors will aid in assessment of the distribution patterns of flea parasitism, and is an important tool in developing control strategies and in evaluation of flea-borne disease risk in dogs and humans. In this survey we used data from 744 domestic dogs from 79 localities in Spain to explore the associations between the abundance of flea species, host-dependent factors (sex and age), and host habitat factors including abode (farm, house with garden, apartment), location (urban or rural), the presence of other pets, and dog activity (measured as the frequency with which dogs left their abode). We also considered environmental factors including the time of year and mean annual temperature and rainfall. Variations in flea community structure at infracommunity and component community levels were also explored. Four flea species were found parasitizing dogs. Ctenocephalides felis was the most abundant (88.02% of fleas identified), followed by Ctenocephalides canis (10.38%), Pulex irritans (1.47%) and Echidnophaga gallinacea (0.13%). Overall flea abundance was higher on dogs living on farms than in apartments, as was the abundance of Ct. felis, Ct. canis and P. irritans. Ct. felis was more abundant on dogs living in houses than in apartments, but the reverse was found for P. irritans. Overall flea abundance and Ct. canis abundance were highest in rural areas, whereas the presence of other pets sharing the abode was associated with higher overall flea abundance and Ct. felis abundance. Only P. irritans abundance was positively related to the activity of dogs. Ct. canis and P. irritans abundances were higher during the warm period of the year. Mean annual temperature was negatively correlated with overall, Ct. canis and P. irritans abundances, but positively related to Ct. felis abundance. Annual rainfall was negatively correlated with Ct. canis and P. irritans abundances. Variations in the number of flea species found on a dog reflected the abundance distribution patterns for each species and their associations with host habitat and environmental factors. At the component community level, flea species richness was inversely related to annual mean temperature. The structure of flea assemblages on dogs was mainly associated with host habitat and environmental variables, and not with host-dependent variables. However, a large amount of variation in flea abundance remained unexplained, suggesting the effect of other non-controlled factors.