Dirofilaria Immitis Research Articles

Evaluation of renal values during treatment for heartworm disease in 27 client-owned dogs

BackgroundCanine heartworm disease (CHD) caused by Dirofilaria immitis remains a common preventable disease with increasing incidence in some parts of the USA. The treatment guidelines of the American Heartworm Society (AHS) currently recommend monthly macrocyclic lactone administration, 28 days of doxycycline given orally every 12 h and three injections of melarsomine dihydrochloride (1 injection on day 2 of treatment followed 30 days later by 2 injections 24 h apart). Minocycline has also been utilized when doxycycline is unavailable. The systemic effects of CHD, which particularly impact cardiac and renal function, have been described, with infected dogs often experiencing renal damage characterized by an increase in serum concentrations of renal biomarkers. Although the AHS treatment protocol for CHD has been shown to be safe and effective in most cases, the potential for complications remains. No study as of yet has evaluated changes in symmetric dimethylarginine (SDMA), a sensitive marker of renal function, during treatment for CHD. The purpose of the present study was to evaluate renal function in dogs by measuring serum creatinine and SDMA concentrations during the adulticide treatment period.MethodsSerum creatinine and SDMA concentrations were measured in 27 client-owned dogs affected by CHD at the following time points: prior to starting doxycycline or minocycline therapy (baseline), during doxycycline or minocycline therapy (interim), at the time of the first dose of melarsomine (first dose), at the time of the second dose of melarsomine (second dose) and at the dog’s follow-up visit after treatment, occurring between 1 and 6 months after completion of therapy (post-treatment). Concentrations of creatinine and SDMA were compared between time points using a mixed effects linear model.ResultsMean SDMA concentrations following the second dose of melarsomine were significantly lower (−1.80 ug/dL, t-test, df = 99.067, t = −2.694, P-Value = 0.00829) than baseline concentrations. There were no other statistically significant differences in the concentration of either biomarker between the baseline and the other time points in CHD dogs undergoing treatment.ConclusionsThe results suggest that the current AHS protocol may not have a substantial impact on renal function.Graphical

The seroprevalence of canine dirofilariosis in dogs in the eastern coastal areas of China

Background and objectivesThe present study was carried out to assess the presence of canine Dirofilaria immitis infection in pet dogs in China. Materials and methodsFrom October 2018 to November 2019, a total of 216 sera were collected from pet hospitals in Shandong, Jiangsu, Shanghai, Zhejiang, and Fujian regions of Eastern China. The sera were tested by using a commercial canine heartworm antibody ELISA test kit. Results70.8% of the pets had significant clinical symptoms resembled to heartworm infection; the overall dirofilariosis positivity found was 12.5% (27/216); Significant positive rates differences were observed between symptomatic and asymptomatic dogs (P < 0.05) (i.e. 15.7% and 4.7% respectively).The prevalence of infection in Shandong Province (15.5%) was the highest among the surveyed areas, but the difference among the geographic regions was not statistically significant (P > 0.05). Furthermore, the prevalence detected in summer (28.2%) was significantly higher than in other seasons (P < 0.05). In addition, no significant difference was observed between male and female sex (P > 0.05). ConclusionsAltogether, these results suggest that an epidemic of dirofilariosis exists in eastern coastal China, as such preventive measures should be taken to control the spread of dirofilariosis to reduce the risk of human and pet infection with heartworm.

Analysis of exotic pathogens found in a large group of imported dogs following an animal welfare investigation.

Increased dog relocation can cause dissemination of pathogen and vector populations, and this is being recognised in countries across Northern Europe, including the UK. Data regarding the prevalence of exotic infections in dogs entering the UK would be beneficial to veterinarians to help assess pets entering the UK from abroad and to help calculate the risk of establishment of novel pathogens. This study reports the findings from a group of imported dogs that was seized as part of a Royal Society for the Prevention of Cruelty to Animals (RSPCA)-led animal welfare investigationand subsequently blood tested for exotic pathogens. As part of the RSPCA investigation, 151 dogs were removed from the site. Blood tests were performed for Babesia canis, Ehrlichia canis, Hepatozoon canis and Leishmania infantum by PCR, Brucella canis by antibody serology and Dirofilaria immitis by blood antigen. In addition to pathogen screening, a serology titre for rabies was measured for each dog. A clinical examination was performed by a veterinary surgeon, and clinical signs were recorded. Overall, 24% (32/133) of the dogs tested positive for an infection with one or more exotic pathogens. Two dogs were positive for Br. canis antibodies and had no clinical signs indicative of infection. Leishmania was identified in 10.5% (14/133) of dogs, and all but two of these were implanted with microchips of Romanian origin. H. canis was identified in 9.6% (10/104) of dogs, all of whom had a Romanian microchip. D. immitis was identified in 4.1% (5/121) of dogs, B. canis in 2.3% (3/129) of dogs and E. canis in only 1.5% (2/131) of dogs tested. Only four dogs were found to have co-infections. No significant association was found between the pathogens detected and presenting clinical signs. This was a group of rescued dogs that were tested for a range of pathogens. They were not randomly selected and as such do not represent the true prevalence of these pathogens in dogs imported into the UK. This study demonstrates a range of exotic pathogens entering the UK, including Br. canis, and demonstrates the importance of screening imported dogs. The emphasis on early recognition of exotic pathogens in imported dogs has relied on screening based on relevant clinical signs and the country of origin. While these factors are useful, this study demonstrated no significant association between presenting clinical signs and the pathogens carried.

Use of In-Clinic Diagnostic Kits for the Detection of Seropositivity to Leishmania infantum and Other Major Vector-Borne Pathogens in Healthy Dogs.

Canine Vector-Borne Diseases (CVBDs) are widespread in Europe and enzootic in many other countries. Though severe illnesses may occur, dogs living in enzootic areas often show vague or no clinical signs of CVBDs. Undiagnosed infections/co-infections in subclinically infected animals favor the spread of CVBDs and increase the risk of transmission to other animals and, in some cases, humans. This study has evaluated the exposure of dogs living in key enzootic countries, i.e., Italy and Greece, to major CVBDs via the use of in-clinic diagnostic kits. Overall, 300 privately owned dogs without/with single mild clinical signs living in different regions of Italy (n. 150) and Greece (n. 150) were included in the study. As part of a clinical examination, a blood sample was collected from each dog and subjected to two serological rapid tests, i.e., the SNAP® 4Dx®Plus (IDEXX Laboratories Inc.) for the detection of antibodies against Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi s.l. and Dirofilaria immitis antigen and the SNAP®Leishmania (IDEXX Laboratories Inc.) for the detection of antibodies against Leishmania infantum. In all, 51 dogs (17%; 95% CI 12.9-21.7) were seropositive to at least 1 pathogen, i.e., 4 in Italy (2.7%; 95% CI 1.4-13.1) and 47 in Greece (31.3%; 95% CI 24-39.4). Dirofilaria immitis antigens were found in 39 dogs (13%; 95% CI 9.4-17.3), while antibodies against Ehrlichia, Anaplasma and Leishmania were detected in 25 (8.3%; 95% CI 5.5-12.1), 8 (2.7%; 95% CI 1.2-5.2) and 5 (1.7%; 95% CI 0.5-3.8) dogs, respectively. None of the dogs tested seropositive for B. burgdorferi s.l. Statistical analyses were performed to evaluate associations between exposure to CVBDs and possible risk factors. The present results indicate that dogs living in enzootic areas may be seropositive for one or more CVBDs in absence of clinical signs. Rapid kits are among first line tools for the detection of CVBDs in clinical settings, as they are cost-effective, straightforward and quick to use. Also, in-clinic tests used herein allowed detection of co-exposure to CVBDs investigated.

Animal and Human Dirofilariasis in India and Sri Lanka: A Systematic Review and Meta-Analysis.

Dirofilariasis is an emerging vector-borne tropical disease of public health importance that mainly affects humans and dogs. Dirofilaria immitis and D. repens are the two well-documented dirofilariasis-causing filarioid helminths of both medical and veterinary concerns in India and Sri Lanka. This systematic review and meta-analysis aimed to describe and summarize the current evidence of dirofilariasis prevalence and distribution in India and Sri Lanka. Interestingly, D. repens is reported to circulate in both dogs (prevalence of 35.8% (95% CI: 11.23-60.69)) and humans (97% of published case reports) in India and Sri Lanka, but D. immitis is reported to be present in the dog populations in India (prevalence of 9.7% (95% CI: 8.5-11.0%)), and so far, it has not been reported in Sri Lanka. This peculiar distribution of D. immitis and D. repens in the two neighbouring countries could be due to the interaction between the two parasite species, which could affect the pattern of infection of the two worm species in dogs and thus influence the geographical distribution of these two filarial worms. In medical and veterinary practice, histopathology was the most commonly used diagnostic technique (31.3%; 95% CI 2.5-60.2%). The low specificity of histopathology to speciate the various Dirofilaria spp. may lead to misdiagnosis. It was identified in this study that several regions of India and Sri Lanka have not yet been surveyed for dirofilariasis. This limits our understanding of the geographical distribution and interspecies interactions of the two parasites within these countries. Parasite distribution, disease prevalence, and interspecies interactions between the vectors and the host should be targeted for future research.

Occurrence of canine subcutaneous dirofilariosis (Dirofilaria repens) in Switzerland

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.

Retrospective analyses of heartworm (Dirofilaria immitis) disease and ectoparasite preventive medication compliance in veterinary practices in the USA

BackgroundCompliance failure with administration of heartworm (HW) disease preventives has been reported as the main contributor to HW disease incidence in medicalized dogs. This study aimed to evaluate purchase compliance with different canine HW preventive products in the USA.MethodsAnonymized transaction data from clinics throughout the USA served as the basis for two retrospective analyses. We first examined the monthly equivalent doses of HW preventive purchases from clinics that had implemented extended-release moxidectin injectables ProHeart® 6 (PH6) and/or ProHeart® 12 (PH12) compared to clinics that prescribed monthly HW preventatives (MHWP) only. In the second analysis, the purchase compliance in practices that dispensed only flea and tick (FT) and HW products separately but did not dispense combination products (dual-therapy practices) was compared to the purchase compliance with the combination product Simparica Trio® (sarolaner, moxidectin, and pyrantel chewable tablets), purchased in clinics having implemented combination therapy in their formulary (combination-therapy practices). In both analyses, the numbers of monthly doses dispensed annually per dog were calculated.ResultsTransaction data from 3,539,990 dogs in 4615 practices were included in the first analysis. In dogs administered PH12 or PH6, the numbers of monthly equivalent doses were 12 and 8.1, respectively. In both clinic types, the average annual number of MHWP doses totaled 7.3. In the second analysis, a total of 919 practices were identified as combination-therapy practices and 434 as dual-therapy-only practices. A total of 246,654 dogs (160,854 dogs in dual-therapy practices and 85,800 dogs in combination-therapy practices) were included in the calculation of the average annual number of monthly doses, which totaled 6.8 (HW preventive products) and 4.4 (FT products) in dual-therapy practices compared to 7.2 months for both FT and HW preventives with Simparica Trio® across both practice types.ConclusionsThe injectable HW preventive PH12 is the only product that provides 12 months of heartworm disease prevention in a single veterinarian-administered injection. When choosing a monthly preventive, the combination therapy was associated with a greater purchase compliance compared with FT and HW products being dispensed separately.Graphical

Angiotensin-converting enzyme 2 in dogs with Dirofilaria immitis

BackgroundInfection by the canine heartworm, Dirofilaria immitis, causes significant cardiopulmonary disease, with progression impacted by increasing parasite numbers and duration of infection. The renin–angiotensin–aldosterone system (RAAS) is an important mediator of cardiac and pulmonary disease. Angiotensin-converting enzyme 2 (ACE2) mitigates the maladaptive effects of angiotensin II by converting it to angiotensin (1-7). We hypothesized that circulating ACE2 activity would be altered in dogs with high heartworm infection intensities relative to dogs without heartworms.MethodsFrozen serum samples (−80 °C) from 30 dogs euthanized at Florida shelters were analyzed for ACE2 activity using liquid chromatography–mass spectrometry/mass spectroscopy and a kinetics approach with and without an ACE2 inhibitor. A convenience sample of 15 dogs without heartworms (HW0) and 15 dogs with > 50 heartworms (HW>50) was included. Heartworm number and microfilariae presence were determined at necropsy. The effects of heartworm status, body weight, and sex on ACE2 were evaluated using regression analysis. Values of P < 0.05 were considered significant.ResultsAll HW0 dogs were D. immitis microfilariae-negative and all HW>50 dogs were D. immitis microfilariae-positive with a median adult worm count of 74 (minimum = 63, maximum = 137). The ACE2 activity of HW>50 dogs (median = 28.2 ng/ml; minimum = 13.6, maximum = 76.2) was not different from HW0 dogs (median 31.9 ng/ml; minimum = 14.1, maximum = 139.1; P = 0.53). The ACE2 activity was higher in dogs with high body weight (median 34.2 ng/ml minimum = 14.1, maximum = 76.2) than in dogs with low weight (median 27.5 ng/ml; minimum = 16.4, maximum = 139.1; P = .044).ConclusionsHeartworm infection did not impact ACE2 activity in shelter dogs with or without heartworms, but heavier dogs had higher ACE2 activity compared to lighter dogs. Comprehensive RAAS evaluation and additional clinical information would aid in understanding how ACE2 activity relates to the entire cascade and clinical status in dogs with heartworm disease.Graphical

Management and outcome of intracardiac heartworms in dogs

BackgroundIntracardiac heartworm (IH) disease is a serious condition that can become life threatening if the patient develops caval syndrome. We aim to describe the management and outcome of IH in dogs evaluated by Medvet’s New Orleans cardiology service from November 2015 to December 2021.MethodsRecords of 27 dogs with IH were examined retrospectively. Follow-up information was obtained from phone conversations with referring veterinarians and owners.ResultsNine of 27 dogs had a previous diagnosis of heartworm disease and were undergoing “slow kill” treatment; 12/27 dogs' heartworm disease was a new diagnosis, and 6/27 had either scheduled or started adulticide therapy. Nine dogs had heartworm extraction. No dogs died during the heartworm extraction procedure. Four of 9 dogs have died (survival time 1; 676; 1815 and 2184 days). One dog died the day after the procedure secondary to continued respiratory distress; the other three died of non-cardiac causes. Five of nine are alive (median follow-up 1062 (range 648–1831) days. Eleven dogs had IH resolution. In 7/11 this occurred while undergoing stabilization for heartworm extraction. In 4/11 heartworm extraction was not recommended because of low IH burden. All dogs with IH resolution were discharged from the hospital. Four of 11 have died (survival time 6; 22, 58 and 835 days), and 6/11 are alive (median follow-up 523 (range 268–2081) days. One was lost to follow-up after 18 days. Five dogs were medically managed. In one of five dogs, extraction was not recommended because of low IH burden. In four of five extraction was recommended but declined. One of five has died (survival 26 days), and four of five are alive (follow-up 155, 371, 935 and 947 days). Two dogs were killed at the time of diagnosis. Fifteen of 27 dogs were considered to have caval syndrome.ConclusionThe results suggest that patients with IH resolution have a good long-term prognosis. Most often IH resolution occurred while the dog was undergoing stabilization for heartworm extraction. When IHs are present, heartworm extraction should still be considered the treatment of choice and recommended as first-line therapy whenever possible.Graphical

Heartworm (Dirofilaria immitis) in carnivores kept in zoos in Texas, USA: risk perception, practices, and antigen detection

BackgroundDirofilaria immitis is the causative agent of heartworm disease in wild and domestic canids, felids, and mustelids. Recent studies demonstrate that additional families in the order Carnivora are also susceptible to infection. Therefore, the objectives of this study were to (1) better understand current practices surrounding heartworm prevention and diagnostics in zoological facilities located in the state of Texas, USA, and (2) assess archival serum samples of carnivores kept in these facilities for the presence D. immitis antigen and/or antibody.MethodsA questionnaire was completed by veterinarians or veterinary technicians representing 10 zoological facilities across Texas. This questionnaire was designed at the taxonomic family level, encompassing the 12 terrestrial carnivore families Ailuridae, Canidae, Eupleridae, Felidae, Herpestidae, Hyaenidae, Mephitidae, Mustelidae, Prionodontidae, Procyonidae, Ursidae, and Viverridae. The second objective was achieved with the use of archival serum samples made available by six zoo facilities.ResultsRisk perception varied across facilities for every family, including among species belonging to Canidae. All facilities used monthly heartworm prevention in canids and felids, with more variation existing in the other families. The use of diagnostic testing and type and route of administration of preventive varied by facility, with oral ivermectin the most commonly used preventive. A total of 217 archival serum samples, belonging to 211 individual animals encompassing 11 families and 39 species, were tested with a commercial heartworm antigen ELISA test, pre- and post-immune-complex dissociation. A subset of samples was also assessed for the presence of feline anti-heartworm antibodies using a commercial ELISA test. Two animals, both of which were Asian small-clawed otters from the same facility, had antigen detected (0.95%).ConclusionsThis study demonstrates that while the zoo veterinary community is aware of the risk and health impact of heartworm disease in canids and felids, there is still a great deal of uncertainty regarding the risks and ideal strategies for prevention in other carnivore families. The low proportion of antigen detection may serve as a baseline for future prevalence studies across the southern United States, where there is an emerging concern of macrocyclic lactone resistance in heartworm.Graphical

Treatment of dogs with Bravecto® (fluralaner) reduces mosquito survival and fecundity

BackgroundMosquitoes serve as the vector of canine heartworm (Dirofilaria immitis), which represents a significant and persistent threat to canine health. A reduction in the longevity and/or reproductive success of mosquitoes that take a blood meal from fluralaner-treated dogs may consequently reduce the local transmission of heartworm and prevent new infections. A novel secondary effect of an oral formulation of the ectoparasiticide fluralaner (Bravecto®) against a laboratory strain of the mosquito Aedes aegypti, a potential major vector of canine heartworm, was investigated in this study.MethodsSix dogs were administered a single dose of fluralaner orally in the form of Bravecto® Chews (at the labeled fluralaner dose of 25 mg/kg body weight), while six control dogs received no treatment. Mosquitoes were fed on blood that was collected from each dog prior to treatment and weekly for 15 weeks post-treatment to assess the continued effects of fluralaner as its serum level decreased. Mosquito fitness was assessed by three parameters: rate of successful blood-feeding, survival, and egg laying.ResultsSuccessful blood-feeding rate was similar between control and treatment groups. In the fluralaner treatment, mosquito survival was significantly reduced within the first 24 h after blood-feeding, for the first 12 weeks post-treatment of the dogs (efficacy range = 33.2–73.3%). Survival of mosquitoes up until a potentially heartworm-infective timepoint (14 days post-blood-feeding) was significantly reduced in the fluralaner-treated group at several timepoints (1, 2, 5, 11, 12, 13, 14, and 15 weeks post-treatment; efficacy range = 49.4–91.4%), but was less consistently reduced at the other timepoints. Egg laying by mosquitoes was almost completely suppressed for the first 13 weeks following treatment of the dogs with fluralaner (treatment efficacy ≥ 99.8%).ConclusionsMosquitoes fed blood from fluralaner-treated dogs experienced a significant reduction in survival and fecundity. These findings support the potential for a reduction in heartworm transmission directly by lethal effects on the vector and indirectly through a reduction of the local vector population when mosquitoes are exposed to animals treated with fluralaner.Graphical

First molecular confirmation of multiple zoonotic vector-borne diseases in pet dogs and cats of Hong Kong SAR

In recent years, the incidence of vector-borne diseases (VBDs) has increased throughout the globe. In particular, tick-borne diseases (e.g., caused by Ehrlichia canis, E. ewingii, Anaplasma phagocytophilum, A. platys, Borrelia burgdorferi sensu stricto (s.s.) and Babesia gibsoni) and mosquito-borne diseases (e.g., caused by Dirofilaria immitis) diseases pose a burden on animal health. Nevertheless, there have been no studies undertaken on the occurrence of VBDs in pet dogs and cats in Hong Kong SAR. This study fills this gap, and is the first to determine the seroprevalence of major VBDs, such as those caused by D. immitis, E. canis, E. ewingii, A. phagocytophilum, A. platys and B. burgdorferi s.s, in dogs and cats through commercially available SNAP 4Dx plus testing. Infection by all these pathogens and Babesia sp. was further assessed through PCR and DNA sequencing. A total of 224 blood samples were collected from domestic dogs (n = 159) and cats (n = 65) in Hong Kong SAR during summer 2022. Hematocrit and platelet counts were determined in each blood sample and other hematological parameters were assessed using an automatic hematology analyzer and vortex the specimen for one to two minutes at or near the highest setting to minimize the clumping. All cat sera samples were negative for tested pathogens, but antibodies against some of the pathogens were detected in dog sera samples. Here, the highest figures were recorded for seroprevalence of E. canis/E. ewingii (10.7%), followed by D. immitis (5.7%), and A. phagocytophilum/A. platys (2.5%). No B. burgdorferi s.s. antibodies were detected in any of the dogs tested. Through molecular diagnostics, we detected the presence of B. gibsoni (3.7%), E. canis (3.1%), D. immitis (5.7%), and A. phagocytophilum (1.3%). Neighbor-Joining phylogenetic trees for vector-borne pathogens (i.e., genus Anaplasma sp.) showed 100% clustering to Japan, the USA and Germany, whereas genus Ehrlichia sp. showed 100% clustering to China, Turkey, Cuba, and Greece. Similarly, genus Babesia sp. clustered 100% to India, Sri Lanka and Austria, while D. immitis clustered in Iraq, South Korea, Portugal, France, the USA and Italy. This study provides the first evidence on the occurrence of tick-borne pathogens in pet dogs in Hong Kong SAR. Based on these findings, it is recommended that appropriate screening should be undertaken in domestic dogs to evaluate the prevalence of these pathogens and promote the timely control of VBDs.

The long-distance relationship between Dirofilaria and the UK: case report and literature review.

Over the last two decades, vector-borne pathogens (VBPs) have changed their distribution across the globe as a consequence of a variety of environmental, socioeconomic and geopolitical factors. Dirofilaria immitis and Dirofilaria repens are perfect exemplars of European VBPs of One Health concern that have undergone profound changes in their distribution, with new hotspots of infection appearing in previously non-endemic countries. Some areas, such as the United Kingdom, are still considered non-endemic. However, a combination of climate change and the potential spread of invasive mosquito species may change this scenario, exposing the country to the risk of outbreaks of filarial infections. Only a limited number of non-autochthonous cases have been recorded in the United Kingdom to date. These infections remain a diagnostic challenge for clinicians unfamiliar with these "exotic" parasites, which in turn complicates the approach to treatment and management. Therefore, this review aims to (i) describe the first case of D. repens infection in a dog currently resident in Scotland, (ii) summarise the available literature on Dirofilaria spp. infections in both humans and animals in the United Kingdom and (iii) assess the suitability of the United Kingdom for the establishment of these new VBPs.

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, non‑stimulated 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.

Disease Surveillance of Cougars (Puma concolor) in Utah, USA.

Adult, free-ranging cougars (Puma concolor) were sampled in three regions of Utah, US, from 2018 to 2021. A total of 68% (23/34) of the sampled cougars had antibodies to feline parvovirus, 15% (5/33) to canine distemper virus, 18% (6/34) to calicivirus, and 22% (8/37) to Yersinia pestis. Forty-one percent (13/32) had IgG antibodies to Toxoplasma gondii and 6% (2/33) to feline immunodeficiency virus, and 3% (1/32) were positive for Dirofilaria immitis (heartworm) antigen. All were seronegative for Toxoplasma gondii IgM, feline enteric coronavirus, SARS-CoV-2, feline leukemia virus, feline herpesvirus, and Francisella tularensis. Tapeworms and Toxascaris leonina eggs were detected in the feces. The disease exposures detected were similar to what has been reported from cougar populations in other western US states, and the current level of exposures is unlikely to have a negative impact on the state's population.

The first molecular confirmation of Culex pipiens complex as potential natural vectors of Dirofilaria immitis in Myanmar.

Dirofilariosis, known as one of the most widespread vector-borne zoonotic diseases, is caused by several different species of the nematodes of the genus Dirofilaria, which can be transmitted by Culex, Anopheles and Aedes mosquito vectors. In order to identify key vector mosquitoes of filarial parasites in Myanmar, mosquitoes were collected during three different seasons (summer, rainy and winter) in three townships in Nay Pyi Taw area, Myanmar. DNA extraction and polymerase chain reaction (PCR) analyses were conducted for 185 mosquito pools, with each pool containing 1-10 mosquitoes. Dirofilaria immitis was detected in 20 pools of Culex pipiens complex mosquitoes. The minimum infection rate of mosquitoes was found to be 16.33. The small subunit ribosomal RNA (12S rDNA) gene targeted PCR revealed that the sequences obtained were completely identical to the sequences of D. immitis derived from dogs in China, Brazil and France. The sequences obtained from mitochondrial cytochrome oxidase subunit I (COI) gene PCR exhibited 100% identity with the sequences of D. immitis derived from dogs in Bangladesh, Iran, Japan and Thailand, as well as humans in Iran and Thailand, and mosquitoes in Germany and Hungary. The findings of this study demonstrated that the mosquito species of Cx. pipiens complex are potential mosquito vectors for dirofilariosis in Myanmar.

Leptospira spp. and Dirofilaria immitis in dogs in Tonga: a pilot study

Dogs are common in Tonga and exist in a close relationship with humans, both as free-ranging pets and guard dogs for domestic properties. Little is known about pathogens present in these animals, and this pilot study aimed to develop a methodology to identify the presence of zoonotic pathogens Leptospira spp. and Dirofilaria immitis (the causative agent of heartworm), in a sample of 82 dogs voluntarily presented to a spey/neuter clinic, using blood samples and point-of-care tests. No positive tests were returned for Leptospira spp. or D. immitis, despite the presence of Leptospira spp. having previously being identified in Tonga.

Parasitism of Dirofilaria incrassata Molin, 1858 in coatis (Nasua nasua).

Populations of the coati, Nasua nasua, like the populations of other wild animals, are regulated by several biotic or abiotic factors. For example, parasites act as a biotic factor affecting the dynamics and density of coati populations. The parasitic nematodes of coatis include Dirofilaria species, such as Dirofilaria immitis, Dirofilaria repens, and Dirofilaria incrassata. Considering that there are few records on parasitism by D. incrassata, including information on the life cycle or location in the host body, this study aimed to investigate the infection of N. nasua by D. incrassata in midwestern Brazil. Thus, two coatis (males and adults) from the Cerrado of Goiás, which died (cause unknown) at the Wild Animal Screening Center/IBAMA in the municipality of Goiânia, Goiás, Midwestern of Brazil, were dissected and all helminths found were collected, identified following specific keys and quantified. A total of 85 specimens of D. incrassata were collected, with a mean parasitic intensity of 42.5 and a parasitic amplitude ranging from 40 to 45, measuring 41-93mm in length and 0.23-0.45mm in width. All helminths were adults and were distributed in the superficial and deep fascia (at different levels) from the neck to the hind limb. Most helminths were tangled, with some encased in a connective tissue film. Most reported human heartworm cases refer to subcutaneous or ocular heartworm infection, mainly caused by D. repens, although other species of heartworm may be associated. D. incrassata was not reported as zoonotic agent, diferently of other Dirofilaria species associated with wild animals and with zoonotic potential in the Americas. This study reiterates N. nasua as the definitive host for D. incrassata and the subcutaneous tissue as the site of choice for this helminth in the adult form. In addition, it reports new body regions where the parasite occurs. This study is the first to document the occurrence of D. incrassata infestation in the State of Goiás, Brazil.

Investigation of comorbidities in dogs with leishmaniosis due to Leishmania infantum

In endemic areas, dogs with leishmaniosis due to Leishmania infantum frequently have comorbidities, including mostly neoplastic, infectious, and parasitic diseases. The aim of this study was to compare the prevalence of comorbidities among dogs that are not infected by L. infantum, dogs that are infected but do not present leishmaniosis, and dogs with leishmaniosis, and to examine if certain comorbidities are independent risk factors for the infection by L. infantum and/or for the development of canine leishmaniosis (CanL). A total of 111 dogs, older than 1-year and non-vaccinated against CanL, were allocated into three groups: group A (n=18) included dogs that were not infected by L. infantum, group B (n=52) included dogs that were infected by L. infantum but did not present CanL, and group C (n=41) included dogs with CanL. Signalment and historical data were obtained using a structured questionnaire. Laboratory examinations included complete blood count, serum biochemistry, urinalysis, fecal parasitology, modified Knott's test, microscopic examination of capillary blood, buffy coat, lymph node, bone marrow and conjunctival smears, qualitative serology for Dirofilaria immitis, Anaplasma phagocytophilum/A. platys, Borrelia burgdorferi and E. canis, IFAT for L. infantum, ELISA for Babesia spp. and Neospora caninum, and real-time PCR for L. infantum in bone marrow, skin biopsies and conjunctival swabs. A variety of comorbidities were found in all three groups. No independent risk factors for infection by L. infantum were found. On the contrary, among dogs infected by L. infantum, being a mongrel [odds ratio (OR): 11.2], not receiving prevention for dirofilariosis (OR: 26.5) and being seropositive to N. caninum (OR: 17.1) or to Babesia spp. (OR: 37.6), were independent risk factors for presenting CanL. Although no comorbidities influence the probability of canine infection by L. infantum, certain comorbidities may be precipitating factors for the transition from the subclinical infection by L. infantum to the overt CanL.

Safety of Simparica Trio® (sarolaner, pyrantel, moxidectin) in heartworm-infected dogs

BackgroundAssessment of the safety of heartworm preventatives in dogs with pre-existing patent heartworm (Dirofilaria immitis) infections is necessary because rapid adult worm and microfilarial death can lead to severe clinical complications, including thromboembolism and anaphylactic shock in dogs. The aim of this study was to determine the clinical safety of Simparica Trio® (sarolaner, pyrantel, moxidectin) in heartworm-infected dogs and the degree of microfilaricidal and adulticidal activity of three consecutive monthly treatments of Simparica Trio.MethodsTwenty-four laboratory Beagle dogs were implanted with 10 male and 10 female D. immitis (ZoeKY isolate), and once infection was patent, they were randomized equally among three groups to receive no treatment, 1× or 3× the maximum recommended label dose of Simparica Trio. Dogs in the treated groups received Simparica Trio on days 0, 28 and 56. In-life assessments included body weight, physical examinations, clinical observations, daily general health observations, a quantitative estimate of food consumption and blood collections for pharmacokinetic (PK) analysis, microfilariae (MF) counts and D. immitis antigen testing. At the end of the study the heart, lungs and pleural and peritoneal cavities were examined for adult D. immitis worms.ResultsSimparica Trio was generally well tolerated. Emesis occurred at low frequency in all groups including control. Abnormal stool occurred occasionally in the 1× and 3× groups throughout the 3-month study. Fever (> 104 °F/40 °C) was recorded in one 1× and one 3× dog 1 day after the first dose and resolved by the following day. No severe hypersensitivity reactions occurred. The mean number of circulating microfilariae (MF) counts in the control group increased from 12,000/ml at study start (Day 0) to > 20,000/ml at Day 28 and remained > 20,000/ml for the duration of the study. The least squares means of circulating MF were reduced by 69.8% on Day 1 and 97.4% on Day 7 for the 1× group and remained at > 99% lower than the control group for the remainder of the study. Similarly, least squares means of circulating MF were reduced by 85.3% on Day 1 and 93.9% on Day 7 for the 3× group and remained > 98% lower than the control group for the remainder of the study. At the end of the study, the mean number of implanted adult worms recovered was < 10 per sex in all groups with 90%, 85% and 75% of live adult heartworms recovered in control, 1× and 3× treatment groups, respectively. Low numbers of dead adult worms were recovered in 1× and 3×, with none in control. Following each dose, the moxidectin and sarolaner AUC and Cmax had close to dose proportional increases.ConclusionsThis study demonstrated that Simparica Trio (sarolaner, pyrantel, moxidectin) was well tolerated when administered to heartworm-positive dogs at 1× and 3× the maximum recommended dose at 28-day intervals for 3 consecutive months. Simparica Trio significantly reduced microfilaria counts in both treatment groups, without significant clinical consequences. At the doses administered, Simparica Trio had minor adulticidal activity but resulted in no clinical sequelae.Graphical