Dirofilaria immitis and Onchocercidae spp. in wild felids from Brazil.

Among the species described within the Onchocercidae family, Dirofilaria immitis is regarded as the most common worldwide, causing severe and often fatal conditions in dogs, cats, and occasionally humans. Dirofilaria spp. are vectored by mosquitoes, simulids, and culicoids, with their epidemiology dependent on the geographical distribution of competent vectors. Eight species of Dirofilaria have been reported so far in Brazil, of which six parasitize non-human primates, deer, procyonids, and marsupials. Here, we investigated the occurrence of Onchocercidae in wild felids (i.e., Panthera onca, Puma concolor, Herpailurus yagouaroundi, Leopardus geoffroyi, Leopardus guttulus, Leopardus pardalis, Leopardus wiedii, Leopardus munoai) from different locations in Brazil. Overall, 82 samples (n = 63 blood; n = 19 tissues) were molecularly screened for cytochrome c oxidase subunit-1 (cox1) gene. Four (i.e., 4.8%) wild felid samples were positive, and at BLAST analysis, the obtained sequences showed varying percentage of nucleotide identity with the genera Brugia (i.e., 87-88%), Setaria (i.e., 89%), and D. immitis (i.e., 94.4%). Phylogenetic analyses clustered sequences obtained into three distinct clades, one with D. immitis and the remaining two with other Onchocercidae spp. Data herein obtained highlight the need for a more comprehensive understanding of the diversity and biology of Onchocercidae in South America in order to assess the potential impact that these species may have for domestic and wild animals, as well as humans.

Evaluation of feline heartworm disease based on gross necropsy, serology, pulmonary histopathology, and radiographic evidence in adult shelter cats in northeastern Alabama.

BACKGROUND: Veterinary knowledge regarding feline heartworm has been increasing significantly over the past two decades. Necropsy surveys of shelter cats have shown feline adult heartworm infection prevalence to be 5-20% of the rate in unprotected dogs; however, other studies have shown feline heartworm antibody prevalence up to 33%, reflecting higher exposure rates and potential immature adult infections. Thus, the true prevalence of feline heartworm infection is likely underestimated due to the limitations of current diagnostic techniques, inadequate testing protocols, and the high likelihood of cats exhibiting transient clinical signs or dying without confirmation of infection. Diagnosing Feline Heartworm Disease (FHWD), also referred to as Heartworm Associated Respiratory Disease (HARD), is one of the conundrums of veterinary medicine. The purpose of this study was to evaluate and characterize the occurrence of Heartworm Associated Respiratory Disease [HARD] in shelter cats, naturally-infected with D.immitis. METHODS: Fifty shelter cats slated for euthanasia between December 2009 and June 2010 were investigated by gross necropsy, radiography, serology, and lung histopathology using techniques that have been established in experimental models of cat heartworm infection. The relationship between pulmonary vascular disease and serological markers for heartworm was also examined using correlations and statistical modeling. Serology included standard heartworm antigen test and a commonly used heartworm antibody test. Also included were heat-treated heartworm antigen test and two additional heartworm antibody tests previously evaluated on experimentally-infected cats. RESULTS: None of the cats were heartworm antibody (HW Ab) positive on a commonly used HW Ab test used by many reference laboratories even though 20% of the study cats were heartworm antigen (HW Ag) positive on heat-treated samples. Two additional HW Ab test were positive on 26% and 22% of the study cats. The combination of heat-treated HW Ag, HW Ab tests, and histopathology indicated 34% of the study cats had HARD. CONCLUSIONS: Utilizing both, the above tests, and thoracic radiographs, enhanced the ability to predict vascular disease, possibly caused by infection with immature and adult heartworms and supported the premise that cats develop heartworm disease at the same rate as dogs.

Genome structure and population genomics of the canine heartworm Dirofilaria immitis.

The heartworm, Dirofilaria immitis, is a filarial parasitic nematode responsible for significant morbidity and mortality in wild and domesticated canids. Resistance to macrocyclic lactone drug prevention represents a significant threat to parasite control and has prompted investigations to understand the genetic determinants of resistance. This study aimed to improve the genomic resources of D. immitis to enable a more precise understanding of how genetic variation is distributed within and between parasite populations worldwide, which will inform the likelihood and rate by which parasites, and in turn, resistant alleles, might spread. We have guided the scaffolding of a recently published genome assembly for D. immitis (ICBAS_JMDir_1.0) using the chromosomal-scale reference genomes of Brugia malayi and Onchocerca volvulus, resulting in an 89.5 Mb assembly composed of four autosomal- and one sex-linked chromosomal-scale scaffolds representing 99.7% of the genome. Publicly available and new whole-genome sequencing data from 32 D. immitis samples from Australia, Italy and the USA were assessed using principal component analysis, nucleotide diversity (Pi) and absolute genetic divergence (Dxy) to characterise the global genetic structure and measure within- and between-population diversity. These population genetic analyses revealed broad-scale genetic structure among globally diverse samples and differences in genetic diversity between populations; however, fine-scale subpopulation analysis was limited and biased by differences between sample types. Finally, we mapped single nucleotide polymorphisms previously associated with macrocyclic lactone resistance in the new genome assembly, revealing the physical linkage of high-priority variants on chromosome 3, and determined their frequency in the studied populations. This new chromosomal assembly for D. immitis now allows for a more precise investigation of selection on genome-wide genetic variation and will enhance our understanding of parasite transmission and the spread of genetic variants responsible for resistance to treatment.

Genetic variability of Dirofilaria repens isolates from humans and dogs in Italy.

Dirofilaria repens is a paradigmatic example of an emerging vector-borne pathogen (VBP) in both human and veterinary fields. The spatial expansion and the increasing zoonotic impact of this VBP can be related to several drivers including the genetic structure of parasite populations. Italy is one of the European countries traditionally endemic with the highest incidence of canine and human cases of subcutaneous dirofilariosis. The present study aimed to assess the genetic identity and variability of D. repens isolates of human and canine origin from areas of Central Italy, compared with those isolated from different areas of Europe by sequence analysis of mtDNA genes (i.e., 12 S rDNA and cox1). A total of twenty isolates of D. repens were obtained from biopsies of subcutaneous and ocular cases of dirofilariosis occurring in 10 dogs and 10 humans. The sequence analysis of 12 S rDNA showed that all the sequences obtained clustered as a monophyletic group with a strong nodal support, indicating that all sequence types represented D. repens. The cox1 and the 12 S sequence analysis did not show host-related polymorphisms between human and dog-derived specimens. The sequence analysis of cox1 was performed including 8 additional sequences previously obtained from human and canine isolates in the same areas. Out of the 28 sequences analyzed, 20 were grouped in a haplogroup comprising 15 haplotypes (i.e., DR1, DR2, DR4, DR5, DR7, DR8, DR10-DR18), 2 sequences matched to DR9, reported for the first time in Italy, and 6 showed peculiar polymorphisms that were not previously described. The results obtained have implications for a better understanding of the epidemiology and phylogeography of this emerging vector-borne zoonotic parasite.

Vector-borne parasites in dogs from Ukraine translocated to Poland following Russian invasion in 2022.

INTRODUCTION: Since 24 February 2022, the day the Russian aggression against Ukraine began, millions of refugees and thousands of pets crossed the Polish-Ukrainian border. Additionally, an unknown number of shelter and stray dogs and cats were rescued and translocated to Poland by private persons and non-profit organizations. The aim of the present study was to examine rescued dogs and cats for presence of canine vector-borne parasites to determine the role of armed conflict in spreading these parasites. METHODS: In July 2022 blood samples were collected from two animal shelters in central Poland hosting dogs and cats rescued from Ukraine. Animals were imported from various regions of Ukraine, including eastern and southeastern Ukraine (military conflict area). Fifty-three dogs (51 from two shelters and two owned ones) and one shelter cat were examined by molecular methods (PCR and sequencing) for the presence of Babesia/Theileria spp., Dirofilaria spp. and Hepatozoon spp. DNA. RESULTS: We detected Dirofilaria immitis, a parasite species non-endemic in Poland, in two dogs translocated from Ukraine (2/53 = 3.8% [95% CI 0.5-13.0%]). One dog had a history of previous heartworm infection. High prevalence of Hepatozoon canis (51% [95% CI 36.8-64.9%]) was noted among translocated dogs. Prevalence of Dirofilaria repens in Ukrainian dogs (18.9% [95% CI 9.4-32.0%) was similar to prevalence in dogs from central Poland (12%). Co-infection of D. repens and D. immitis was found in two dogs and six dogs were co-infected by D. repens and H. canis. Hepatozoon canis infection was also identified in an imported cat. CONCLUSION: We confirmed that this military conflict has facilitated the spread of canine vector-borne parasites, including zoonotic species.

First molecular characterization of Dirofilaria Immitis in Cuba.

BACKGROUND: Dirofilarioses are widespread diseases caused by mosquito-borne nematodes of the family Onchocercidae, genus Dirofilaria. The major etiologic agent of canine dirofilariosis in the American continent is the zoonotic parasite Dirofilaria immitis. Existing reports of filarioid nematodes in Cuba are based solely on morphological and immunological analysis which do not allow unambiguous identification and/or direct detection of causal agents. RESULTS: Here we present the molecular characterization of filarioid nematodes found in a dog in Cuba. Based on the molecular and phylogenetic analysis of the 5.8S-ITS2-28S region and cox1 gene fragments, the worms were unambiguously classified as D. immitis. Sequence analysis showed high identity of the gene fragments in this study with others previously obtained from D. immitis found in dogs, wolfs and jackals but also from mosquito vectors of D. immitis. CONCLUSIONS: Further studies are guarantee to better understand the epidemiological impact of canine dirofilariosis in Cuba as well as the competence of different species of culicid mosquitoes as vectors of Dirofilaria in the country.

Association between vector-borne pathogen seroprevalence in shelter-housed and owned dog populations in the contiguous United States of America.

Domestic dogs are susceptible to numerous vector-borne pathogens that are of significant importance for their health. In addition to being of veterinary importance, many of these pathogens are zoonotic and thus may pose a risk to human health. In the USA, owned dogs are commonly screened for exposure to or infection with several canine vector-borne pathogens. Although the screening data are widely available to show areas where infections are being diagnosed, testing of owned dogs is expected to underestimate the actual prevalence in dogs that have no access to veterinary care. The goal of this study was to measure the association between the widely available data from a perceived low-risk population with temporally and spatially collected data from shelter-housed dog populations. These data were then used to extrapolate the prevalence in dogs that generally lack veterinary care. The focus pathogens included Dirofilaria immitis, Ehrlichia spp., Anaplasma spp., and Borrelia burgdorferi. There was a linear association between the prevalence of selected vector-borne pathogens in shelter-housed and owned dog populations and, generally, the data suggested that prevalence of heartworm (D. immitis) infection and seroprevalence of Ehrlichia spp. and B. burgdorferi are higher in shelter-housed dogs, regardless of their location, compared with the owned population. The seroprevalence of Anaplasma spp. was predicted to be higher in areas that have very low to low seroprevalence, but unexpectedly, in areas of higher seroprevalence within the owned population, the seroprevalence was expected to be lower in the shelter-housed dog population. If shelters and veterinarians make decisions to not screen dogs based on the known seroprevalence of the owned group, they are likely underestimating the risk of exposure. This is especially true for heartworm. With this new estimate of the seroprevalence in shelter-housed dogs throughout the USA, shelters and veterinarians can make evidence-based informed decisions on whether testing and screening for these pathogens is appropriate for their local dog population. This work represents an important step in understanding the relationships in the seroprevalences of vector-borne pathogens between shelter-housed and owned dogs, and provides valuable data on the risk of vector-borne diseases in dogs.

Evaluation of Anaplasma spp. seroprevalence in dogs and association with incidence of human anaplasmosis.

Point-of-care (POC) ELISA tests are routinely used in US veterinary practices to screen canine patients for antibodies to tick-transmitted pathogens. Results are also used to monitor spatial and temporal trends in canine seroprevalence, and these data can build awareness of the risk to humans of tick-transmitted diseases such as Lyme disease and anaplasmosis. This study utilized a second-generation test that has incorporated additional Anaplasma-specific peptides into a commercial POC ELISA test to allow detection of Anaplasma spp. antibodies earlier post-infection. A convenience population consisting of 19,894 canine samples from a US commercial diagnostic laboratory were tested using the second-generation POC ELISA test to describe regional Anaplasma spp. canine seroprevalence and assess correlation to anaplasmosis cases reported to Centers for Disease Control and Prevention by state. Antibodies to Anaplasma spp. were detected in 1646 samples (8.3%) with the Northeast and Midwest US census regions having the highest proportion of positive samples. At the state level, a significant correlation was found between canine Anaplasma spp. seroprevalence and human anaplasmosis incidence (r2 = 0.64). Although estimates of canine Anaplasma spp. seroprevalence presented here using the second-generation POC ELISA are generally increased, especially in the Northeast and Midwest, the regional distribution of canine samples testing positive for Anaplasma spp. antibodies is consistent with previous reports. The observed correlation with human anaplasmosis incidence indicates that results from the second-generation POC ELISA will continue to add value in epidemiological assessment of human anaplasmosis risk.

Dirofilaria immitis conquering the regions in Slovakia previously endemic for D. repens.

The present study was focused on the current state of Dirofilaria species distribution in the territory of the Slovak Danubian Lowland, a region previously identified endemic for Dirofilaria repens. For the research, blood samples of 330 dogs tested positive for dirofilariosis using concentration tests or "rapid heartworm tests" were sent by private veterinary practitioners for further DNA analyses and species determination. The results revealed an unquestionable change in the pattern of Dirofilaria species distribution with Dirofilaria immitis, diagnosed as the agent of mono- and co-infections with D. repens, responsible for 60.00% of all cases. The results showed that in the course of the last 5 years, D. immitis has spread significantly in Slovakia and has become the dominant causal agent of dirofilariosis in the former endemic areas of D. repens distribution, which increases infection risk for both dogs and humans.

Dirofilaria immitis in pet dogs from the metropolitan area of the Colombian Caribbean.

A cross-sectional study was carried out to determine the frequency and factors associated with Dirofilaria immitis infection in pet dogs in the metropolitan area of the Colombian Caribbean (northern Colombia). A total of 173 dogs were analyzed by a commercial rapid immunochromatographic test (RIT) and a nested PCR of the cytochrome oxidase subunit I gene, in parallel. Ninety-two (53.2%) of the dogs showed positive results to the RIT, while 59 (34.1%) animals had D. immitis DNA by PCR. Positivity to one or both diagnostic techniques was detected in 104 (60.1%; CI95%: 53.8-67.4) of the sampled dogs. In PCR-positive dogs, phylogenetic analyses evidenced high nucleotide identity (100%) with sequences previously obtained from mosquitoes, dogs and other mammals in different countries. Exercise intolerance (p = 0.002; OR: 2.33; CI95%: 1.37-3.96) and thrombocytopenia (p = < 0.001; OR: 1.95; CI95%: 1.11-3.43) were the main factors associated with D. immitis infection in dogs. The high frequency of D. immitis in dogs indicates a wide distribution of this parasite in the metropolitan area of the Colombian Caribbean, which can be of animal and public health concern. Our results highlight the need to combine different methods to increase the diagnostic accuracy of D. immitis.

The first report of Dirofilaria repens infection in dogs from Colombia.

Dirofilariasis is a mosquito-borne disease caused by Dirofilaria parasites, affecting both wild and domestic animals, including humans considered as accidental hosts. Dirofilaria repens is the principal causative agent of dirofilariasis in the Old World, with increasing reports of the parasite in countries where it has not been previously identified, due to several factors such as the expansion of mosquito vectors' geographical distribution. By utilizing newly designed primers for molecular detection and confirming through next-generation sequencing, here, we report the first plausible cases of D. repens in dogs from Colombia. Our results support the classification of this species as an emergent pathogen in the Americas. Finally, we encourage an increase in diagnostic and surveillance efforts to prevent and control the current and future dirofilariasis cases in this region.

Elimination of Dirofilaria immitis Infection in Dogs, Linosa Island, Italy, 2020-2022.

On Linosa Island, Italy, Dirofilaria immitis infection has been hyperendemic in dogs and seroprevalent among islanders. In 2020, we implemented a heartworm disease elimination program on Linosa Island. Of 54 dogs tested for D. immitis antigen and microfilariae, 28 had positive results and received treatment with oral doxycycline twice daily for 4 weeks plus topical imidacloprid/moxidectin monthly for 12 months. The 26 dogs with negative results received monthly topical imidacloprid/moxidectin as preventive. During month 1, the number of microfilaremic dogs was reduced by 76.5%. From month 2 on, all animals were microfilariae negative, and during months 3 to 9, the number of antigen-positive dogs decreased progressively. Treatment of positive dogs coupled with chemoprophylaxis for noninfected dogs was effective, protecting them from new infections. The elimination program reduced the risk for human infection, representing a One Health paradigm. Monitoring and chemoprophylaxis are advocated to maintain the status of heartworm disease-free area.

A rare subcutaneous periorbital zoonotic infection in a 42-year-old female patient.

Human dirofilariasis represents a zoonotic infectious disorder caused by parasites belonging to the Dirofilaria genus, which includes numerous species with a large variability regarding the host specificity, life cycle, and clinical manifestations. This disease appears to be a vector-borne parasitosis that is spread through insects - intermediate hosts (usually mosquitos) - and affects several carnivores as definitive hosts. Humans represent an unsuitable host for the parasite to complete its life cycle, being unable to release microfilariae in the blood as the inoculated larvae fail to reach sexual maturity. Therefore, humans are unable to transmit the infection to other humans, neither directly nor through an intermediate host. The current case report indicates a rare encounter of a Dirofilaria repens infection in a 42-year-old female patient living in an urban area (Craiova, Romania), who developed intermittent right periorbital edema after a previous trip to Greece. Over the course of one month, the right periorbital edema gradually remitted, and a firm, round lump developed in the external inferior right periorbital region. There were no similar clinical manifestations among other family members. Her medical history showed no other complaints or any serious general health problems. Also, the patient denied having any pets at home. The blood tests did not reveal any abnormalities. The exact source of infection could not be identified, but the chances for this infection to be related to the history of traveling to Greece are high, given that this region is one of the most important endemic areas in Europe.

Dirofilariosis infestation of pectoralis major muscle in Italy, a case report.

Abstract: Zoonotic dirofilariasis infestation, caused by Dirofilaria Repens, is described worldwide in different countries. A 31-years-old male patient presented thoracic muscle pain after growth of an ovoidal undefine cyst in left parasternal region. Patient reported several contacts with different species of animals for a familiar activity. In absence of blood inflammatory indices and systemic symptoms, imaging studies showed a suspected muscle cyst infection. Surgical excision was performed and microbiology confirmed parasite nature. Dirofilaria Repens, probably adult female, was identified. Treatment resulted to be definitive and any other clinical and surgical approach was needed. Healing time was uneventful and follow-up showed no further systemic relapses. The case highlights the effectiveness of surgical treatment in this subcutaneous infestation for an increasing number of human cases reported in endemic areas such as Central Italy.

The prevalence of canine dirofilariasis in China: a systematic review and meta-analysis.

BACKGROUND: Dirofilariasis, the disease caused by Dirofilaria spp., and in particular by Dirofilaria immitis and Dirofilaria repens in canines, occurs frequently in canids and felids, and occasionally in humans, in temperate, sub-tropical and tropical regions globally. Although highly effective, safe and convenient preventive medicines have been available for the treatment of dirofilariasis for the past three decades, the disease remains a major veterinary and public health concern in endemic areas. The insect vectors, host-parasite relationships and interactions of Dirofilaria spp. have received little attention in China, and there is very little information in English regarding the prevalence of dirofilariasis in animals and humans in the country. The aim of this systematic review and meta-analysis is to evaluate the status of canine dirofilariasis in China based on the available literature in English and in Chinese. METHODS: We systematically searched five databases for epidemiologic studies on the prevalence of canine dirofilariasis in China and finally selected 42 studies eligible for inclusion in the systematic review and meta-analysis. The meta-analysis was performed using the random effects model in the meta package in R v4.2.1. RESULTS: The random effects model gave a pooled and weighted prevalence of Dirofilaria infection among dogs in China in the past 100 years of 13.8% (2896/51,313, 95% confidence interval 8.2-20.4%) with a high level of heterogeneity (I2 =  99.5%). CONCLUSIONS: Our analyses indicated that the prevalence of canine dirofilariasis in China has gradually declined, but that the range of Dirofilaria spp. has expanded. Older and outdoor dogs presented a higher rate of positive infection. The findings indicated that more attention should be paid to host factors for the effective control and management of this disease.

CANINE HEARTWORM INFECTION AND PROPHYLAXIS USE AMONG PET CARETAKERS FROM THE CUMBERLAND GAP REGION OF TENNESSEE, USA.

The prevalence of infection with canine heartworm (CHW), Dirofilaria immitis, continues to increase across the United States, regardless of the availability of effective and affordable prophylactic products. Current reports of CHW prevalence as estimated by the Companion Animal Parasite Council (CAPC) are thought to under-represent the true magnitude of the issue because pet dogs that do not receive regular veterinary care are often excluded. This study estimated the prevalence of CHW in pet dogs and associated prophylaxis use in pet dogs in the Cumberland Gap Region with a combined doorstep diagnostic testing approach and caretaker survey. Dogs tested (n = 258) during the summers of 2018 and 2019 revealed a 2.3% (6/258) prevalence in the pet dog population with 33% (2/6) being microfilaremic. Questionnaire data from caretaker interviews revealed that 41.8% (108/258) of the dogs were not receiving CHW prophylaxis. Significant predictors of CHW prophylaxis use identified through logistic regression included pet caretaker awareness of CHW as an important health issue and the use of veterinary services in the year preceding participation in the survey. These results underscore the importance of veterinary-mediated client interaction to create risk awareness of CHW disease and association with prophylaxis compliance.

Spatial and risk factor analyses of vector-borne pathogens among shelter dogs in the Eastern United States.

BACKGROUND: Vector-borne infections pose significant health risks to humans, domestic animals, and wildlife. Domestic dogs (Canis lupus familiaris) in the United States may be infected with and serve as sentinel hosts for several zoonotic vector-borne pathogens. In this study, we analyzed the geographical distribution, risk factors, and co-infections associated with infection with Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi, and Dirofilaria immitis in shelter dogs in the Eastern United States. METHODS: From 2016 to 2020, blood samples from 3750 shelter dogs from 19 states were examined with IDEXX SNAP® 4Dx® Plus tests to determine the seroprevalence of infection with tick-borne pathogens and infection with D. immitis. We assessed the impact of factors including age, sex, intact status, breed group, and location on infection using logistic regression. RESULTS: The overall seroprevalence of D. immitis was 11.2% (n = 419/3750), the seroprevalence of Anaplasma spp. was 2.4% (n = 90/3750), the seroprevalence of Ehrlichia spp. was 8.0% (n = 299/3750), and the seroprevalence of B. burgdorferi was 8.9% (n = 332/3750). Regional variation in seroprevalence was noted: D. immitis (17.4%, n = 355/2036) and Ehrlichia spp. (10.7%, n = 217/2036) were highest in the Southeast while seroprevalence for B. burgdorferi (19.3%, n = 143/740) and Anaplasma spp. (5.7%, n = 42/740) were highest in the Northeast. Overall, 4.8% (n = 179/3750) of dogs had co-infections, the most common of which were D. immitis/Ehrlichia spp. (1.6%, n = 59/3750), B. burgdorferi/Anaplasma spp. (1.5%, n = 55/3750), and B. burgdorferi/Ehrlichia spp. (1.2%, n = 46/3750). Risk factors significantly influenced infection across the evaluated pathogens were location and breed group. All evaluated risk factors were significant for the seroprevalence of D. immitis antigens. CONCLUSIONS: Our results demonstrate a regionally variable risk of infection with vector-borne pathogens in shelter dogs throughout the Eastern United States, likely due to varying distributions of vectors. However, as many vectors are undergoing range expansions or other changes in distribution associated with climate and landscape change, continued vector-borne pathogen surveillance is important for maintaining reliable risk assessment.

[Occurrence of canine subcutaneous dirofilariosis (Dirofilaria repens) in Switzerland]. / Vorkommen des Hautwurms Dirofilaria repens bei Hunden in der Schweiz.

INTRODUCTION: Infectious diseases are increasingly introduced into Switzerland due to the increased travel activity in recent decades and the import of dogs. Dirofilariasis, caused by Dirofilaria immitis or D. repens, is one of them. An infection with D. repens, the cause of canine subcutaneous dirofilariosis, is often asymptomatic in dogs, but represents a potential zoonotic disease risk for humans. Due to the rapidly increasing number of human cases, D. repens is considered an emerging zoonosis in north-eastern Europe. The prevalence of D. repens infections in dogs and humans in Switzerland is unknown. Since 2016 the analysing diagnostic laboratory provided with the newly introduced filaria PCR a realiable diagnostic test to differentiate betweeen D. immitis and D. repens. Total nucleic acid (DNA and RNA) was extracted from 200 µl EDTA blood without prior enrichment followed by a species-specific real-time PCR assay. The analyses for Dirofilariae in the years 2016 to 2021 were examined in a descriptive retrospective study and the proportion of positive tests per year (prevalence with 95 % confidence interval) was calculated. Furthermore, blood samples of 50 imported dogs to Switzerland were analyzed in an exploratory cross-sectional study for the presence of dirofilaria. No D. repens positive case was found in the first two years after the introduction of the PCR. In 2018 five of 546 analyzed samples (5/546, 0,9 %, 95 % confidence interval [95 % CI]=0,7 - 1,2 %) tested positive for D. repens, in 2019 four of 591 (0,7 %, 95 % CI=0,5 - 0,9 %), in 2020 15 of 783 (1,9 %, 95 % CI=1,6 - 2,3 %) and in 2021 eleven of 1058 samples (1,0 %, 95 % CI=0,8 - 1,3 %). In the exploratory cross-sectional study, four of the 50 examined dogs were positive for D. repens (8 %, 95 % CI=2,6 - 20,1 %). One dog had a concurrent infection with D. immitis and D. repens. All four positive tested dogs were imported from Hungary. Potentially zoonotic infections with D. repens occur in dogs living in Switzerland. This disease should be included in the differential diagnoses of imported dogs and a monitoring during routine health checks should be done more often. The veterinary profession can thus assume its responsibility for the prevention of zoonoses as part of a One Health approach.

INTRODUCTION: Au cours des dernières décennies, des maladies infectieuses sont de plus en plus souvent introduites en Suisse en raison de l'augmentation des voyages et de l'importation de chiens. La dirofilariose, causée par Dirofilaria immitis ou D. repens, est une de ces maladies. Une infection par D. repens, responsable de la dirofilariose sous-cutanée canine, est souvent asymptomatique chez le chien, mais elle représente un risque potentiel de zoonose pour l'homme. En raison de l'augmentation rapide du nombre de cas humains, D. repens est une zoonose émergente dans le nord-est de l'Europe. La prévalence des infections à D. repens chez les chiens et les humains en Suisse est inconnue. L'introduction de la PCR des filaires dans le laboratoire d'analyse permet depuis 2016 de faire une distinction fiable entre D. immitis et D. repens. L'acide nucléique total (ADN et ARN) a été extrait de 200 µl de sang EDTA sans enrichissement préalable, suivi d'un test PCR en temps réel spécifique à l'espèce. Les résultats des analyses de la dirofilariose de 2016 à 2021 ont été examinés dans une étude rétrospective descriptive et la proportion de tests positifs par an (prévalence avec un intervalle de confiance de 95 %) a été calculée. En outre, des échantillons de sang de 50 chiens importés en Suisse ont été analysés dans le cadre d'une étude transversale exploratoire pour la présence de dirofilaires. Aucun cas positif à D. repens n'a été trouvé au cours des deux premières années suivant l'introduction de la PCR. En 2018, cinq des 546 échantillons analysés (5/546, 0,9 %, intervalle de confiance à 95 % [95 %CI]=0,7­1,2 %) ont été testés positifs pour D. repens, en 2019 quatre des 591 (0,7 %, CI 95 %=0,5 ­ 0,9 %), en 2020 15 des 783 (1,9 %, CI 95 %=1,6 ­ 2,3 %) et en 2021 onze des 1058 échantillons (1,0 %, CI 95 %=0,8 ­ 1,3 %). Dans l'étude transversale exploratoire, quatre des 50 chiens examinés étaient positifs pour D. repens (8 %, CI à 95 % = 2,6 ­ 20,1 %). Un chien présentait une infection concomitante par D. immitis et D. repens. Les quatre chiens testés positifs avaient été importés de Hongrie. Des infections potentiellement zoonotiques par D. repens se produisent chez des chiens vivant en Suisse. La surveillance de cette maladie devrait être davantage incluse dans les contrôles de santé de routine et dans les diagnostics différentiels chez les chiens importés. La profession vétérinaire peut ainsi assumer sa responsabilité dans la prévention des zoonoses dans le cadre d'une approche One Health.

Dirofilaria immitis is endemic in rural areas of the Brazilian Amazonas state capital, Manaus.

The canine filarial parasite Dirofilaria immitis has not been reported in Brazil´s Amazonas state capital, Manaus, for over a century. Here, we report one imported and 27 autochthonous D. immitis infections from a microfilarial survey of 766 domestic dog blood samples collected between 2017 and 2021 in Manaus. An Overall prevalence estimate of 15.44% (23/149) was calculated from our two rural collection sites; a prevalence of 1.22% (4/328) was estimated at our periurban collection site, and an overall prevalence of 0.35% (1/289) was calculated from our two urban clinic collections. Our data suggest that in the urban areas of Manaus, where the parasites are very likely vectored by the same species of mosquito that historically vectored Wuchereria bancrofti (Culex quinquefasciatus), prevalence levels are very low and possibly maintained by an influx from rural areas where sylvatic reservoirs and/or more favorable vector transmission dynamics maintain high prevalences.

Occurrence of Dirofilaria immitis infection in shelter cats in the lower Rio Grande Valley region in South Texas, United States, using integrated diagnostic approaches.

Dirofilaria immitis is a mosquito-borne filarioid nematode that affects dogs and cats. Although heartworm infections in cats can be fatal, it is commonly neglected by cat owners and veterinarians. Moreover, diagnosing heartworm infection in cats can be challenging, requiring the integration of multiple laboratorial tests and clinical examination. The objective of this study was to estimate the occurrence of D. immitis infection in shelter cats in the Lower Rio Grande Valley (RGV) region of Texas using a combination of immunodiagnostic and molecular methods. The RGV has a large population of stray animals with limited access to veterinary care. One hundred and twenty-two paired samples of serum and DNA extracted from the blood clots of cats from 14 towns in this region were analyzed. Serum samples were used for heartworm antibody detection (Heska® Solo Step®), and heartworm antigen detection using a commercial ELISA kit (DiroCHEK®) pre- and post-immune-complex dissociation (ICD) via heat treatment. A species-specific probe-based qPCR assay targeting a fragment of the cytochrome oxidase c subunit 1 of the mitochondrial DNA was utilized to detect the presence of parasite DNA. Twenty-two cats (18%) were positive in at least one diagnostic test. Antibody testing detected most cases (19/122; 15.6%); pre- and post-ICD antigen testing detected 6 cases (6/122; 4.9%); and qPCR detected the fewest cases (4/122; 3.3%), with 2 cats positive on all three diagnostic tests. Veterinarians should encourage local cat owners to utilize year-round heartworm prevention.