Current State of Canine Heartworm in Portugal.

The favourable geo-climatic conditions in Portugal have made it highly conducive to the development of Dirofilaria immitis in dogs, leading to its identification as an endemic region. This nematode is rapidly spreading across Europe, particularly in northeastern countries. The objective of this study was to provide an updated assessment of the prevalence of this disease in Portuguese dogs, analysing the results in relation to epidemiological and geo-environmental factors, and to identify potential risk factors. A total of 1367 dogs from all continental and insular districts were included in the study and tested for D. immitis antigens. The overall prevalence was found to be 5.9%. It was observed that the disease is spreading northward, with previously unaffected districts now reporting cases, and that the prevalence in coastal districts exceeded that of inland ones. Notably, the Aveiro district exhibited a significant increase in D. immitis prevalence, while in certain districts such as Setúbal, Santarém, Madeira, or Faro, a stabilisation or decrease in prevalence was noted. Furthermore, outdoor and older dogs were found to be at a higher risk of infection. No positive cases were detected in the Azores. Most of the infected dogs were located in areas of high and medium risk of infection. This study underscores the importance of implementing pharmacological prophylaxis, vector control strategies, and public awareness programs to control the spread of this zoonotic disease.

Usefulness of Tissue Doppler Imaging for the Evaluation of Pulmonary Hypertension in Canine Heartworm Disease.

BACKGROUND: Dirofilaria immitis is a nematode that produces proliferative pulmonary endarteritis in dogs due to direct contact of the adult parasites with the intima layer of the pulmonary arteries, leading to irreversible severe structural damage and sustained pulmonary hypertension (PH), which can produce severe cardiorespiratory disorders. The purpose of this study was to assess the diagnostic value of the echocardiography tissue Doppler imaging (TDI) in determining the presence of PH in dogs with heartworm disease. METHODS: There were 116 heartworm-infected dogs with PH and 33 healthy dogs included in the study. Based on the right pulmonary artery distensibility index (RPADi) < 29.5%, PH was present in 47.4% of infected dogs. Additionally, the animals were evaluated using other standard alternative echocardiographic measures to estimate PH. Moreover, a total of eight echocardiographic measurements were analysed using the TDI to determine its usefulness in diagnosing PH (E', A', S, E':A', global TDI, HRI-IVCT, HRI-IVRT, R-TEI). RESULTS: The TDI measurements showed significant differences between dogs with and without PH, demonstrating a positive correlation with respect to the RPADi. In addition, cut-off values for the detection of PH with excellent sensitivity and specificity were found for E':A', global TDI, HRI-IVCT, HRI-IVRT and R-TEI. CONCLUSIONS: The TDI mode may be useful as an adjunct diagnostic method for the determination of PH in dogs with Dirofilaria immitis.

Assessment Heartworm Disease in the Canary Islands (Spain): Risk of Transmission in a Hyperendemic Area by Ecological Niche Modeling and Its Future Projection.

Heartworm disease is a vector-borne zoonotic disease caused by Dirofilaria immitis. The Canary Islands (Spain), geolocated close to the coast of Western Sahara, is an archipelago considered hyperendemic where the average prevalence in domestic dogs is high, heterogeneous, and non-uniform. In addition, Culex theileri has been reported as a vector of the disease on two of the most populated islands. Our aim was to develop a more accurate transmission risk model for dirofilariosis for the Canary Islands. For this purpose, we used different variables related to parasite transmission; the potential distribution of suitable habitats for Culex spp. was calculated using the ecological niche model (ENM) and the potential number of generations of D. immitis. The resulting model was validated with the geolocation of D. immitis-infected dogs from all islands. In addition, the impact of possible future climatic conditions was estimated. There is a risk of transmission on all islands, being high in coastal areas, moderate in midland areas, and minimal in higher altitude areas. Most of the dogs infected with D. immitis were geolocated in areas with a high risk of transmission. In 2080, the percentage of territory that will have been gained by Culex spp. is small (5.02%), although it will occur toward the midlands from coastal areas. This new model provides a high predictive power for the study of cardiopulmonary dirofilariosis in the Canary Islands, as a hyperendemic area of the disease, and can be used as a tool for its prevention and control.

Diversity and geographic distribution of haplotypes of Dirofilaria immitis across European endemic countries.

BACKGROUND: Dirofilaria immitis, also known as heartworm, is one of the most important parasitic nematodes of domestic dogs, causing a potentially serious disease, cardiopulmonary dirofilariosis, which can be lethal. This species seems to be less 'expansive' than its sister species Dirofilaria repens, and it is believed that climate change facilitates the spread of this parasite to new non-endemic regions. METHODS: In total, 122 heartworm isolates were analysed from nine endemic countries in Europe (Portugal, Spain, Italy, Greece, Hungary, Romania, Slovakia, and Ukraine) and a single isolate from Bangladesh by amplification and sequencing of two mitochondrial (mt) DNA markers: cytochrome c oxidase subunit 1 (COI) and dehydrogenase subunit 1 (NADH). The main aim of the current study was to determine the genetic diversity of D. immitis and compare it with D. repens haplotype diversity and distribution. DNA was extracted from adult heartworms or microfilariae in blood. Most isolates originated from dogs (Canis lupus familiaris) while 10 isolates originated from wildlife species from Romania, including eight isolates from golden jackals (Canis aureus), one isolate from a Eurasian otter (Lutra lutra) and one isolate from a red fox (Vulpes vulpes). RESULTS: Median spanning network analysis was based on the combined sequence (1721 bp) obtained from two mt markers and successfully delineated nine haplotypes (Di1-Di9). Haplotype Di1 was the dominant haplotype encompassing 91 out of the 122 sequences (75%) from all nine countries and four host species. Haplotype Di2 was the second most common haplotype, formed solely by 13 isolates from Italy. The remaining sequences were assigned to Di3-Di9 haplotypes, differing by 1-4 SNPs from the dominant Di1 haplotype. There was evidence for geographical segregation of haplotypes, with three unique haplotypes associated with Italy and four others associated with certain countries (Di4 and Di7 with Slovakia; Di8 with Greece; Di6 with Hungary). CONCLUSION: Diversity in D. immitis mt haplotypes was lower by half than in D. repens (9 vs. 18 haplotypes in D. immitis and D. repens, respectively), which may be associated with the slower expansion of heartworm in Central and NE Europe. NADH gene appears to be conserved in Dirofilaria sp. by showing lower genetic diversity than the analysed COI gene.

Usefulness of NT-proBNP in dogs with heartworm: could this biomarker be useful to evaluate pulmonary hypertension?

BACKGROUND: In recent years, the usefulness of echocardiography and serum biomarkers in the diagnosis of pulmonary hypertension (PH) in dogs with heartworm disease has been studied. Previously, N-terminal pro B-type natriuretic peptide (NT-proBNP) has shown high concentrations in dogs with heart disease and/or PH as well as its usefulness as a prognostic indicator, but it has never been evaluated in the diagnosis and prognosis of PH in dogs with heartworm disease. The aim was to evaluate the serum concentrations of NT-proBNP in dogs infected by Dirofilaria immitis to determine its usefulness as a tool to detect precapillary PH. METHODS: NT-proBNP was determined in 50 heartworm-infected dogs. Presence/absence of PH was determined echocardiographically, using the Right Pulmonary Artery Distensibility Index (RPAD Index) and the systolic flow of tricuspid regurgitation mainly, together with other echocardiographic measurements following the guidelines of the American College of Veterinary Internal Medicine (ACVIM) for the diagnosis of PH. Other epidemiological parameters (breed, age, sex, status: client-owned or shelter dogs) and clinical parameters (microfilaremia, parasite burden, presence of symptoms, body condition) were collected as well. RESULTS: Moderate-severe PH was present in 40% of the dogs (RPAD Index < 29.5%), NT-proBNP concentrations being significantly higher compared with dogs that did not have PH. A cutoff for NT-proBNP of ≥ 1178.45 pmol/l showed a sensitivity of 64.3% and a specificity of 95.5% for the presence of moderate-severe PH. Older dogs and dogs from shelters showed significantly higher NT-proBNP concentrations. Dogs with symptoms and low body condition presented significantly higher NT-proBNP concentrations as well. CONCLUSIONS: The determination of NT-pro-BNP concentration can be a useful tool in the diagnostic work-up of dogs with heartworm disease and associated PH and can help to identify animals in the more advanced stage of this disorder.

Feline heartworm disease and environmental allergens hypersensitivity: is there a link?

BACKGROUND: Cats can be infected by Dirofilaria immitis, the causative agent of heartworm disease, characterized by respiratory signs, airway hyperreactivity, remodelling and inflammation. Allergy is a multifactorial pathology, and the role of a number of helminth parasites in the development of allergies in humans and other species has been demonstrated in many studies. The aim of the present study was to verify whether cats seropositive for D. immitis present hypersensitivity to some environmental allergens. METHODS: Blood samples were collected from 120 cats and tested for the presence of specific immunoglobulin G antibodies against D. immitis and for hypersensitivity to 20 allergens, using commercial allergen test kits. RESULTS: Of the 120 cats tested, 72 (60.0%) were seropositive for anti-D. immitis IgG and 55 (45.8%) showed clinical signs of heartworm disease of a respiratory nature. The results of testing with the allergen kits showed that 50.8% of cats were seropositive for ≥ 1 allergens, with the most common allergens being Dermatophagoides farinae (25.8%), Dermatophagoides pteronyssinus (20.0%), Malassezia (17.5%) and Ctenocephalides felis (14.2%). The prevalence of allergies was significantly higher-by almost threefold-in cats seropositive for D. immitis (68.1% vs. 25%). There were no significant differences between the prevalence of allergic cats and presence/absence of symptoms, and the results confirmed that symptoms were not a decisive factor for the presence of allergies. The risk for developing allergies was 6.3-fold higher in cats seropositive for D. immitis than in cats that were seronegative, confirming that seropositivity for D. immitis is a risk factor. CONCLUSIONS: Cats with confirmed heartworm can develop serious respiratory signs, potentially leading to progression to permanent lung injury and predisposing cats to hyperresponsive airway disease. Previous studies have shown that seropositivity for D. immitis and Wolbachia is related to the presence of bronchoconstriction and bronchospasm in the affected cat. The results support the suspicion that contact with D. immitis may be a risk factor for the presence of allergies.

Involvement of the excretory/secretory and surface-associated antigens of Dirofilaria immitis adult worms in the angiogenic response in an in-vitro endothelial cell model.

Angiogenesis is a process by which new vessels are formed from pre-existing ones when the physiological conditions of the vascular endothelium are altered. Heartworm disease, caused by Dirofilaria immitis, causes changes in the vascular endothelium of the pulmonary arteries due to obstruction, friction, and hypoxia. The aim of this study was to analyze whether the excretory/secretory and surface-associated antigens of adult worms interact and modulates the angiogenic mechanism, viable cell number and cell migration, as well as the formation of pseudo-capillaries. Cultures of human vascular endothelial cells (HUVECs) stimulated with excretory/secretory antigens (DiES), surface-associated antigens (Cut) from D. immitis adult worms, VEFG-A (Vascular Endothelial Growth Factor A), as well as DiES+VEFG-A and Cut+VEFG-A were used. The production of VEFG-A and other proangiogenic [soluble VEFGR-2 (sVEFGR-2), membrane Endoglin (mEndoglin)] and antiangiogenic [VEFGR-1/soluble Flt (sFlt), soluble Endoglin (sEndoglin)] molecules was assessed using commercial ELISA kits. Cell viability was analyzed by live cell count and cytotoxicity assays by a commercial kit. In addition, viable cell number by MTT-based assay, cell migration by wound-healing assay carrying out scratched wounds, and the capacity of pseudo-capillary formation to analyze cell connections and cell groups in Matrigel cell cultures, were evaluated. In all cases, non­stimulated cultures were used as controls. DiES+VEFG-A and Cut+VEFG-A significantly increased the production of VEFG-A and sVEFGR-2, and only Cut+VEFG-A significantly increased the production of VEFGR-1/sFlt compared to other groups and non-stimulated cultures. Moreover, only DiES+VEFG-A produced a significant increase in viable cell number and significant decrease cell migration, as well as in the organization and number of cell connections. Excretory/secretory and surface-associated antigens of adult D. immitis activated the angiogenic mechanism by mainly stimulating the synthesis of proangiogenic factors, and only excretory/secretory antigens increased viable cell number, activated cell migration and the formation of pseudo-capillaries. These processes could lead to vascular endothelial remodeling of the infected host and favor the long-term survival of the parasite.

Effect of somatic antigens of Dirofilaria repens adult worms on angiogenesis, cell proliferation and migration and pseudo-capillary formation in human endothelial cells.

BACKGROUND: Angiogenesis is defined as the formation of new vessels by sprouting of endothelial cells from pre-existing vessels in response to stimuli, such as hypoxia or inflammation. Subcutaneous dirofilariasis, caused by Dirofilaria repens, is a zoonotic disease characterized by the formation of subcutaneous nodules with the presence of at least one encapsulated worm, showing perivascular vascularization around it. The aim of this study is to analyze whether the somatic antigen of adult D. repens worms interacts with and modulates the angiogenic mechanism, cell proliferation and migration, and formation of pseudo-capillaries. METHODS: The expression of VEGF-A, VEGFR-1/sFlt, VEGFR-2, mEnd and sEnd in cultures of human vascular endothelial cells stimulated with somatic antigen of adult worms of D. repens (DrSA), vascular endothelial growth factor (VEGF) and DrSA + VEGF were evaluated by using ELISA commercial kits. Cellular viability was analyzed by live cell count, cytotoxicity assays by using a commercial kit, cell proliferation by MTT-based assay, cell migration by wound-healing assay carried out by scratching wounds and capacity of formation of pseudo-capillaries analyzing cell connections and cell groups in Matrigel cell cultures. In all cases unstimulated cultures were used as controls. RESULTS: DrSA + VEGF significantly increased the expression of VEGF-A, VEGFR-2 and mEndoglin compared to other groups and unstimulated cultures. Moreover, DrSA + VEGF produced cell proliferation and migration and increased the formation of pseudo-capillaries. CONCLUSIONS: Somatic antigen of adult D. repens worms activated the proangiogenic mechanism, cell proliferation and cell migration as well as formation of pseudo-capillaries in this in vitro human endothelial cell model. These processes could be related to the survival of adult D. repens in subcutaneous nodules in infected hosts.

Echocardiographic Assessment of the Pulmonary Vein to Pulmonary Artery Ratio in Canine Heartworm Disease.

Background:Dirofilaria immitis produces proliferative pulmonary endarteritis and pulmonary thromboembolism in infected dogs. The pulmonary vascular lesions lead to irreversible and persistent structural damage and, as a consequence, sustained precapillary pulmonary hypertension (PH). The purpose of this study was to assess the diagnostic value of the pulmonary vein to pulmonary artery ratio (PV:PA ratio) to determine moderate or severe PH (>50 mmHg) in dogs with heartworm disease. METHODS: A total of 151 naturally heartworm-infected and 66 healthy dogs were included in the study. The presence/absence of PH was based on the right pulmonary artery distensibility index (RPAD index < 29.5%), and the PV:PA ratio was echocardiographically measured by the time-motion mode (M mode) and two-dimensional mode (2D mode). Other echocardiographic parameters were also assessed (pulmonary trunk to aorta ratio, tricuspid regurgitation pressure gradient, and AT:ET ratio). RESULTS: The results of the PV:PA ratio showed a highly positive correlation between the M and 2D modes (r = 0.928). The PV:PA ratio obtained by the M mode was identified as the strongest predictor for RPAD index (R2 0.628, p < 0.0001) with a good diagnostic accuracy (AUC = 0.99). The results of PV/PA by the 2D mode showed a similar prediction for the RPAD index (R2 0.606, p < 0.0001) with a good diagnostic accuracy (AUC = 0.98). Both of the 2D and M modes' PV:PA ratios decreased significantly with the presence of PH. A cut-off value of ≤0.845 showed high sensitivity and specificity for the M mode (97% and 94%, respectively) and the 2D mode (96% and 93%, respectively). CONCLUSIONS: The PV:PA ratio may be useful as a complementary diagnostic method for the estimation of moderate or severe PH in dogs with heartworm.

Right Ventricle Strain Assessed by 2-Dimensional Speckle Tracking Echocardiography (2D-STE) to Evaluate Pulmonary Hypertension in Dogs with Dirofilaria immitis.

Echocardiographic assessment of the right ventricle is helpful for analysing the pathophysiology of heartworm disease and detecting pulmonary hypertension (PH) in dogs. In veterinary cardiology, the study of myocardial deformation using two-dimensional speckle tracking (2D-STE) echocardiography has become increasingly acknowledged as useful for quantifying right ventricular function. The aim of this study was to evaluate the usefulness of myocardial deformation strain of the right ventricular free wall (FWS), global deformation strain of the right ventricle, including the interventricular septum (GS), and tissue motion annular displacement of the tricuspid valve (TMAD) in a cohort of dogs with heartworm (Dirofilaria immitis) disease and to determine cut-off values for detecting the presence of PH. Out of the 93 dogs tested, 71% were diagnosed with heartworm infection. PH was identified in 41% of the infected dogs following the American College of Veterinary Internal Medicine (ACVIM) guidelines, based on the peak tricuspid regurgitation velocity to calculate the tricuspid regurgitation pressure gradient (TRPG), while other routine measurements were used, including the right pulmonary artery distensibility index (RPADi). The 2D-STE mode measurements were determined using Right Ventricular Automated Function Imaging (RV AFI®) software. The statistical analysis showed significant differences in the studied parameters among dogs with and without PH. Additionally, sensitivity (sen) and specificity (sp) cut-off values were obtained (GS ≥ -21.25%, sen 96%, sp 86.4%; FWS ≥ -21.95%, sen 92.56%, sp 95.5%; TMAD ≤ 0.85 cm, sen 70.4%, sp 83.3%). These results demonstrated that GS, FWS, and TMAD could be used as supplementary and alternative variables to conventional echocardiographic measurements when detecting PH in dogs with heartworm disease.

Comprehensive Map of Canine Angiostrongylosis in Dogs in Spain.

Canine angiostrongylosis is an emerging disease caused by Angiostrongylus vasorum, mainly affecting wild carnivores and dogs. In Spain, there are studies reporting infections in foxes, wolves, and badgers in different regions of the country. However, there are hardly any publications on its prevalence in dogs. The aim of this study was to complete and update the epidemiologic map of A. vasorum in dogs in Spain. A total of 5619 canine blood samples from all autonomous cities and provinces of Spain were collected and tested for the presence of circulating A. vasorum antigens. The overall apparent prevalence of canine A. vasorum infection in Spain was 1.39%. No significant differences were found for sex or age, but significant differences between outdoor and indoor/outdoor dogs were found. A high prevalence was also observed in the northern third of the country, where an oceanic climate prevails, being humid and rainy and where abundant vegetation can be found, thus favoring the proliferation of intermediate hosts. The results suggest that A. vasorum canine infections are heterogeneously present in a large part of the territory, demonstrating its expansion throughout the country, and therefore, awareness and prevention campaigns for this disease should be promoted.

Evaluation of Pulmonary Hypertension in Dogs with Heartworm Disease Using the Computed Tomographic Pulmonary Trunk to Aorta Diameter Ratio.

Dirofilaria immitis causes proliferative pulmonary endoarteritis that leads to the appearance of chronic precapillary pulmonary hypertension (PH) in dogs. Pulmonary trunk to aorta ratio (PT:Ao ratio) obtained by computed tomography (CT) was studied and the quantitative measure of the diameters of the pulmonary trunk (PT), the descending thoracic aorta (DAo) and ascending thoracic aorta (AAo) were evaluated for the determination of the presence of moderate to severe PH in 59 dogs. The diagnosis of PH was echocardiographically determined, based on the determination of the right pulmonary artery distensibility (RPAD) index (<29.5%), and compared with other parameters for estimating PH. The results showed a very high concordance: 0.976 (p-value 0.000) between the two CT methods (PT:DAo and PT:AAo) with an excellent intraclass correlation coefficient > 0.95. Moreover, cut-off values of ≥1.111 for PT:DAo, and ≥1.057 for PT:AAo were determined for dogs with an RPAD index < 29.5%, which suggests a cut-off value between healthy dogs and the presence of PH. As has been previously published, The PT:Ao ratios did not determine the presence of mild PH, so the measurements cannot be considered useful for the early diagnosis of PH in dogs with heartworm.

What Has Happened to Heartworm Disease in Europe in the Last 10 Years?

Heartworm disease caused by Dirofilaria immitis is a vector-borne disease that affects canids and felids, both domestic and wild, throughout the world. It is a chronic disease which causes vascular damage in pulmonary arteries, and in advanced stages, the presence of pulmonary hypertension and right-sided congestive heart failure can be evidenced. Moreover, pulmonary thromboembolism is caused by the death of the worms, which can be lethal for the infected animal. Furthermore, it is the causative agent of human pulmonary dirofilariosis, being a zoonotic disease. The aim of this review was to update the current epidemiological situation of heartworm in Europe in dogs, cats, wild animals, and vectors insects, and to analyse the factors that may have contributed to the continuous spread of the disease in the last decade (2012-2021). In Europe, the disease has extended to eastern countries, being currently endemic in countries where previously only isolated or imported cases were reported. Furthermore, its prevalence has continued to increase in southern countries, traditionally endemic. This distribution trends and changes are influenced by several factors which are discussed in this review, such as the climate changes, presence of vectors in new areas, the appearance of new competent vector species in the continent, increased movement of pets that travelled to or originated from endemic countries, the urbanisation of rural areas leading to the formation of so-called "heat islands", or the creation of extensive areas of irrigated crops. The continuous expansion of D. immitis must be monitored, and measures adapted to the situation of each country must be carried out for adequate control.

Dirofilaria immitis: Genotyping Randomly Selected European Clinical Samples and USA Laboratory Isolates with Molecular Markers Associated with Macrocyclic Lactone Susceptibility and Resistance.

Dirofilaria immitis is a parasitic nematode and causes dirofilariosis, a potentially fatal pulmonary infection which primarily infects canids. Dirofilariosis infections are controlled via prophylactic macrocyclic lactone (ML) regimens. Recent evidence has confirmed the development of ML-resistant isolates in the USA, which are genetically distinct from wildtype populations. Single nucleotide polymorphisms (SNP) associated with ML-resistant phenotypes were clinically validated in USA populations. In this study, 3 USA laboratory-maintained isolates (Berkeley, Georgia II, and WildCat) and 11 randomly selected European clinical samples from 7 hosts were analyzed. The samples tested were fresh microfilaria (mf) in blood or adult worms preserved in ethanol. The samples underwent MiSeq sequencing of the top 9 SNP associated with ML resistance. The results provide the first genotypic analysis of the three USA laboratory-maintained isolates and any European samples. The European clinical samples show no genomic evidence of ML resistance based on the 9 SNP. The early adoption of genotyping of clinical D. immitis samples could provide an early indication of the potential development of ML resistance and aid to distinguish clinical cases of heartworm infection due to ML resistance from those due to a lack compliance with the recommended treatments, as has been seen in North America.

Seroprevalence of Feline Heartworm in Spain: Completing the Epidemiological Puzzle of a Neglected Disease in the Cat.

Feline heartworm is a vector-borne zoonotic disease caused by Dirofilaria immitis. It is a cosmopolitan disease that is continuously expanding. Spain is considered an endemic country; however, although there are many published studies in dogs, feline heartworm has been poorly studied in this country. Thus, the objective was to analyze the exposure to D. immitis throughout Spain to complete the epidemiological map in the feline species. For this, 6,588 feline serum samples were analyzed for the presence of D. immitis antigens and antibodies against D. immitis and Wolbachia. The results were analyzed according to sex, age, breed, habitat, origin (owned or shelter cats), presence of clinical signs, use of preventive, location and climatology. The results showed a prevalence of 0.5% and a seroprevalence of 9.4%. The highest antibody seroprevalences were reported in the Canary Islands and the Balearic Islands (19.2 and 16%, respectively), as well as in the autonomous communities located on the Mediterranean coast (9.2-11.2%). Seropositive cats were found in both indoor and outdoor cats, and from 6 months of age. Furthermore, only 5.8% of cats received regular prophylactic treatment. The results show that feline dirofilariasis is widely distributed throughout the national territory and corroborate that, where infected dogs are present, there are cats exposed to the parasite. It is necessary to implement efficient awareness and prophylaxis measures to control the incidence and expansion of feline heartworm in Spain.

Expansion of Canine Heartworm in Spain.

The climate of Spain has favourable characteristics for the development of D. immitis in dogs, being an endemic country. Given that vector-borne diseases are spreading rapidly through Europe, due to factors such as climate change, the expansion of vectors and the increased mobility of reservoir animals, the aim was to update the epidemiology of heartworm in dogs and analyse the results based on climate and other epidemiological and geo-environmental factors. To this aim, 9543 blood samples from dogs from all provinces and autonomous cities of Spain were analysed for the detection of antigens of D. immitis, obtaining a prevalence of 6.47%. The northwestern and southern provinces showed the highest prevalences, as well as in the Balearic and Canary Islands. Prevalences were higher in dogs outdoors. Furthermore, most of the positive dogs were found in regions with high humidity and water availability. This study shows, for the first time, positive cases in provinces and islands where no cases had previously been described and demonstrates the continuous expansion and consolidation of heartworm in Spain. Considering its zoonotic character, the implementation of control and awareness programmes for the prevention of D. immitis in pets is necessary.

Serological Survey of Canine Vector-Borne Infections in North-Center Spain.

Various factors are currently causing an increase in vector-borne parasitic diseases at a global scale; among them, some stand out, such as climatic disturbances derived from global change, the increase in movements of reservoir animals, or changes in land made by human activity. In the European continent, there have been an increasing number of epidemiological studies focused on the detection of these diseases, especially in dogs. In Spain, there are few epidemiological studies focused on the evaluation of the biotic and abiotic factors that may influence the distribution, such as climatic zones, orography, or presence of water reservoirs. The aim of this study was to analyze the prevalence and distribution of several canine vector-borne diseases caused by Dirofilaria immitis, Leishmania infantum, Anaplasma platys, and Ehrlichia canis in the autonomous community of Castilla y León, the largest region of the Iberian Peninsula, providing a geospatial approach based on a geographic information system (GIS) analysis. Blood from a total of 1,475 domestic dogs from the nine provinces of Castilla y León were analyzed. Also, a GIS analysis of the sample locations was carried out, taking into account the most important predictor variables. The prevalence in dogs infected by D. immitis was 7.19%, and the seroprevalence by L. infantum was 4.61 and 1.56% for A. platys and E. canis. Most of the infected animals were located in areas with stagnant water, irrigated agriculture, or riverbanks, always close to forest and woodland vegetation. These results indicate that dogs living in Castilla y León should take prophylactic measures to avoid infections.

Angiostrongylus vasorum in Domestic Dogs in Castilla y León, Iberian Peninsula, Spain.

Angiostrongylus vasorum is the causative agent of canine angiostrongylosis, a disease affecting domestic and wild canids. In Europe, it is an emerging disease, mainly reported in red foxes. In Spain, there are a few studies that address the prevalence and pathology of this disease. Castilla y León is the largest region of the Iberian Peninsula, whose extensive area is 94,224 km2; however, until now, there have been no epidemiological studies on this disease. Therefore, the aim of this study was to analyze the presence of antigens of A. vasorum in 1475 dogs from Castilla y León, showing an overall prevalence of 0.75%. The infected dogs were mainly outdoors, guard and hunting breed dogs and living in locations with mild climates close to areas of high edaphic humidity, such as stagnant water, irrigated crops or riverbanks, with the vegetation dominated by alders, holm oak and gall oak forests, where the intermediate hosts develop. It is necessary to carry out more in-depth studies on the epidemiology and pathology of this disease in Spain and Europe in order to carry out efficient control in both domestic and wild animals.

Current Distribution of Selected Vector-Borne Diseases in Dogs in Spain.

Currently, climate change, modifications of landscapes and habitats due to human activities, as well as an increase in the movement of reservoirs and new species of competent vectors, have contributed to the spread of canine vector-borne diseases. These are mostly emerging and neglected diseases, some of them with zoonotic potential. Therefore, the objective of this study was to assess the prevalence and distribution of four major canine vector-borne diseases (Dirofilaria immitis, Leishmania infantum, Anaplasma spp., and Ehrlichia canis) in Spain. Between September 2018 and February 2020, blood was sampled from 4643 client-owned dogs from 111 veterinary clinics from the 17 autonomous communities of Spain. All samples were tested for the detection of D. immitis antigens, and for antibodies against L. infantum, Anaplasma spp. and E. canis. Of the studied dogs, 22.14% were positive for one or several diseases while the prevalence was 6.25% (CI: 5.59-6.98) for D. immitis, and the seroprevalences were 10.36% (CI: 9.52-11.27) for L. infantum, 5.06% (CI: 4.47-5.73) for Anaplasma spp., and 4.26% (CI: 3.72-4.88) for E. canis. Co-infections by two and three vector-borne diseases were reported in 13% and 2% of the infected dogs, respectively. The studied vector-borne diseases are widely distributed throughout the Spanish geography, being observed and expanding northward in the case of D. immitis and L. infantum. The results point to an insufficiency of preventive measures to avoid the infection, and the need of the implementation of awareness campaigns among veterinarians and owners. Furthermore, a close collaboration between veterinarians, physicians and health authorities would be necessary for such zoonotic vector-borne diseases.