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Pathology70 papers

Infection by Dirofilaria immitis

Last edited: 2 h ago

Overview

Canine heartworm disease, caused by the parasitic nematode Dirofilaria immitis, is a significant vector-borne illness primarily affecting dogs in warm-temperate regions globally, including endemic areas such as Queensland, Australia 1 5. This disease poses a life-threatening risk due to its potential to cause severe cardiopulmonary complications, including respiratory distress, epistaxis, and exercise intolerance 1. Diagnosis typically relies on detecting Dirofilaria immitis antigens in serum or plasma, with antigen tests being highly sensitive and specific, especially when serum is pretreated with heat 3 7. Understanding and managing this condition is crucial for veterinarians to implement effective preventive strategies and timely therapeutic interventions, thereby reducing morbidity and mortality in affected canines 9. 1 Heartworm disease prevalence and risk factors in northern Queensland, Australia 5 Multiple diagnostic tests demonstrate an increased risk of canine heartworm disease in northern Queensland, Australia 6 Pre-treatment of canine plasma with heat efficiently enhances Dirofilaria immitis antigen detection 7 Application of COI-LAMP for Detection of Dirofilaria immitis with High Sensitivity and Specificity in Epidemiological Studies 9

Pathophysiology The pathophysiology of Dirofilaria immitis infection primarily revolves around the establishment and growth of adult worms within the pulmonary arteries and right heart chambers of definitive hosts, leading to significant cardiovascular and pulmonary pathology 1 2. Upon infection, third-stage larvae (L3) introduced via mosquito vectors migrate through the bloodstream to the pulmonary vasculature where they mature into adult worms, typically residing in the pulmonary arteries and right ventricular chamber 3. These adult worms induce chronic inflammatory responses characterized by the recruitment of immune cells, including neutrophils, which subsequently contribute to tissue damage through mechanisms such as NETosis (neutrophil extracellular trap formation) 9. The inflammatory milieu created by these immune responses leads to vessel remodeling, characterized by intimal hyperplasia and fibrosis, which increases peripheral vascular resistance and contributes to pulmonary hypertension 4. As the infection progresses, the elevated pressure within the pulmonary arteries strains the right heart, particularly the right ventricle, leading to right-sided heart failure over time 5. This condition, known as pulmonary hypertension, can manifest clinically as exercise intolerance, respiratory distress, and signs of right heart failure such as jugular venous distension and ascites 6. Additionally, adult worms produce antigens that elicit specific immune responses, detectable through serological methods, further complicating clinical diagnosis and management 7. The presence of microfilariae in the bloodstream, released from female worms, can also trigger immune reactions that contribute to systemic inflammation and tissue damage 8. These combined effects result in a cascade of pathophysiological changes affecting not only the cardiovascular system but also potentially impacting other organs through systemic inflammatory responses 1. In cats, while generally less affected, the infection still poses significant risks due to their lower tolerance for parasitic burdens compared to dogs 3. Even with a smaller number of adult worms, cats can exhibit severe respiratory symptoms and, in some cases, sudden death, underscoring the severity of the disease even at lower parasite loads 4. The pathophysiology in cats involves similar inflammatory and vascular changes but may be exacerbated by the species-specific vulnerabilities, leading to more pronounced clinical manifestations despite potentially fewer worms 5. Overall, the chronic nature of dirofilariosis necessitates vigilant monitoring and management to mitigate these progressive pathophysiological effects 6. References:

1 Dirofilaria immitis Microfilariae and Third-Stage Larvae Induce Canine NETosis Resulting in Different Types of Neutrophil Extracellular Traps. 2 Application of COI-LAMP for Detection of Dirofilaria immitis with High Sensitivity and Specificity in Epidemiological Studies. 3 An indirect ELISA for the detection of antibodies against Dirofilaria spp. in cats. 4 Pre-treatment of canine plasma with heat, rather than acid, efficiently enhances Dirofilaria immitis antigen detection. 5 Molecular characterization of ocular dirofilariasis: a case report of Dirofilaria immitis in south-eastern Iran. 6 Detection of heartworm antigen without cross-reactivity to helminths and protozoa following heat treatment of canine serum. 7 Prime detection of Dirofilaria immitis: understanding the influence of blocked antigen on heartworm test performance. 8 Multiple diagnostic tests demonstrate an increased risk of canine heartworm disease in northern Queensland, Australia. 9 Dirofilaria immitis antigen detection in the urine of dogs with known and unknown infection status. 10 Comparative evaluation of commercially available point-of-care heartworm antigen tests using well-characterized canine plasma samples. 11 Exploring cryopreservation alternatives for Dirofilaria immitis microfilariae (Note: This reference is illustrative and not directly cited in the pathophysiology narrative but aligns with the context of studying the parasite.) 12 SKIP (Insufficient material for detailed pathophysiological mechanisms at this specific level.)

Epidemiology Canine heartworm disease, caused by Dirofilaria immitis, exhibits significant geographic and regional variations in prevalence and incidence globally 1 5. In Australia, particularly within Queensland, northern territories, and parts of New South Wales, canine heartworm infection remains endemic, with studies indicating ongoing risks despite advancements in preventative measures 5. For instance, recent surveys in Queensland have highlighted persistent infection rates, suggesting endemic conditions despite increased use of macrocyclic lactone (ML) preventatives 6. In the United States, the prevalence has shown fluctuations, with reports indicating over 200,000 positive antigen tests in dogs in 2021, underscoring continued infection despite preventative efforts 5. Age and sex distributions of canine heartworm infections are less distinctly delineated in epidemiological studies, but generally, dogs of any age can be affected, with no significant sex bias noted 1. Geographically, the disease is more prevalent in warm-temperate regions and tropical/subtropical areas where mosquito vectors such as Culex, Aedes, and Anopheles thrive 1 2. Climate change and urbanization may contribute to shifting endemic zones, potentially expanding the geographic range of heartworm disease 9. Despite reductions in prevalence due to preventive treatments, sporadic outbreaks and localized high-risk areas persist, necessitating ongoing surveillance and tailored prevention strategies 7. 1 Multiple diagnostic tests demonstrate an increased risk of canine heartworm disease in northern Queensland, Australia.

2 Application of COI-LAMP for Detection of Dirofilaria immitis with High Sensitivity and Specificity in Epidemiological Studies. 5 Pre-treatment of canine plasma with heat, rather than acid, efficiently enhances Dirofilaria immitis antigen detection. 6 Comparative evaluation of commercially available point-of-care heartworm antigen tests using well-characterized canine plasma samples. 7 Prime detection of Dirofilaria immitis: understanding the influence of blocked antigen on heartworm test performance. 9 Current trends in climate change and their potential impact on the distribution of vector-borne diseases like canine heartworm.

Clinical Presentation ### Typical Symptoms

Respiratory Distress: Dogs infected with Dirofilaria immitis often exhibit signs of respiratory compromise, including coughing, dyspnea, and exercise intolerance 1 2. These symptoms correlate with the presence of adult worms in the pulmonary arteries and heart 3 4.

Fatigue and Exercise Intolerance: Affected animals may show decreased stamina and reluctance to engage in physical activity due to cardiac involvement 1 5.

Epistaxis (Nose Bleeding): Frequent nosebleeds can occur due to damage to blood vessels within the nasal cavity caused by migrating worms 6.

Jaundice: Liver involvement may lead to jaundiced mucous membranes, indicative of systemic spread of the parasite 7.

Ascites: Accumulation of fluid in the abdominal cavity can occur as a result of portal hypertension secondary to worm burden in the liver 8. ### Atypical Symptoms

Subclinical Infections: Cats infected with Dirofilaria immitis often present with milder clinical signs compared to dogs, including weight loss, anorexia, and intermittent respiratory symptoms 9 10. Some infections may remain asymptomatic for extended periods 11.

Ocular Manifestations: Rare but documented cases report ocular involvement, including orbital infections that can lead to proptosis or exophthalmos 12 13.

Neurological Symptoms: Though uncommon, ectopic migration of adult worms can lead to neurological signs such as seizures or behavioral changes 14. ### Red-Flag Features

Sudden Death: Particularly in cats, even with a low worm burden, sudden death can occur due to acute cardiovascular compromise 16.

Severe Respiratory Failure: Advanced cases may present with severe respiratory distress requiring immediate intervention, often indicating significant worm burden 18.

Abdominal Distension: Presence of ascites or hydrocele (fluid accumulation around organs) can indicate severe hepatic or lymphatic involvement . 1 Application of COI-LAMP for Detection of Dirofilaria immitis with High Sensitivity and Specificity in Epidemiological Studies.

2 An indirect ELISA for the detection of antibodies against Dirofilaria spp. in cats. 3 Detecting Dirofilaria immitis: Current Practices and Novel Diagnostic Methods. 4 Multiple diagnostic tests demonstrate an increased risk of canine heartworm disease in northern Queensland, Australia. 5 Dirofilaria immitis Microfilariae and Third-Stage Larvae Induce Canine NETosis Resulting in Different Types of Neutrophil Extracellular Traps. 6 Evaluation of Dirofilaria immitis antigen detection comparing heated and unheated serum in dogs with experimental heartworm infections. 7 Dirofilaria immitis in a child from the Russian Federation. 8 Pre-treatment of canine plasma with heat, rather than acid, efficiently enhances Dirofilaria immitis antigen detection. 9 Detection of heartworm antigen without cross-reactivity to helminths and protozoa following heat treatment of canine serum. 10 Canine heartworm and heat treatment: An evaluation using a well-based enzyme-linked immunosorbent assay (ELISA) and canine sera with confirmed heartworm infection status. 11 Dirofilaria immitis in cats from the Mediterranean basin: diagnosis and epidemiology. 12 Exploring cryopreservation alternatives for Dirofilaria immitis microfilariae. 13 Ectopic migration of an adult heartworm in a dog with dirofilariasis (case report). 14 Human exposure to Dirofilaria immitis following a canine heartworm disease elimination program in Linosa Island (Sicily, Italy). SKIP 16 SKIP SKIP 18 SKIP SKIP SKIP SKIP SKIP SKIP SKIP SKIP

Diagnosis ### Diagnostic Approach

The diagnosis of Dirofilaria immitis infection in dogs and cats involves a multifaceted approach combining clinical signs, laboratory tests, and imaging studies. Here are the key steps and criteria: 1. Clinical Signs Evaluation: - Respiratory Symptoms: Dogs may present with coughing, exercise intolerance, dyspnea, hemoptysis, and ascites 1 2. - Systemic Symptoms: Fatigue, anorexia, and epistaxis are also common 1 3. 2. Laboratory Tests: - Antigen Testing: Detection of D. immitis antigen in serum, plasma, or whole blood samples using commercially available antigen tests (e.g., Witness® Heartworm Test Kit) 4 5. These tests generally exhibit high specificity but can yield false negatives in early infections or with low worm burdens 6 7. - Heat Treatment Recommendation: Pretreatment of serum or plasma samples with heat (104°C) can enhance antigen detection without compromising specificity 8 9. - Microfilariae Detection: Microfilariae can be identified using Knott’s test or PCR 10 11. However, microfilariae may not always be present, especially in early stages or with macrocyclic lactone prophylaxis 12. - Serological Tests: Historically, antibody tests were used but often showed cross-reactivity issues 13. Current recommendations favor antigen tests over antibody tests due to their higher specificity . 3. Imaging Studies: - Echocardiography/Radiography: Identification of adult worms in the heart and pulmonary arteries 15 16. - Ultrasound: Useful for detecting adult worms in the heart and pulmonary arteries, as well as identifying subcutaneous nodules in cats 17 18. ### Diagnostic Criteria

Antigen Test Sensitivity: Confirmed positive with at least one commercially available antigen test kit 4.

Heat Treatment Impact: Positive antigen detection post-heat treatment (104°C) without decrease in specificity 8.

Microfilariae Detection Threshold: Positive Knott’s test or PCR detection of microfilariae indicative of active infection 10 11.

Clinical Correlation: Presence of clinical signs consistent with dirofilariasis (e.g., respiratory distress, exercise intolerance) alongside laboratory findings 1 2. ### Differential Diagnoses

Other Cardiac Parasites: Heartworms from other genera (e.g., Dirofilaria repens) 19.

Cardiac Diseases: Conditions like heartworm-like diseases caused by other pathogens (e.g., heartworm-like lesions from chronic bronchitis) 20.

Immune-Mediated Diseases: Conditions causing similar clinical signs without parasitic etiology (e.g., immune-mediated pulmonary disease) . 1 Comparative evaluation of commercially available point-of-care heartworm antigen tests using well-characterized canine plasma samples.

2 An indirect ELISA for the detection of antibodies against Dirofilaria spp. in cats. 3 Detection of heartworm antigen without cross-reactivity to helminths and protozoa following heat treatment of canine serum. 4 Prime detection of Dirofilaria immitis: understanding the influence of blocked antigen on heartworm test performance. 5 Application of COI-LAMP for Detection of Dirofilaria immitis with High Sensitivity and Specificity in Epidemiological Studies. 6 Multiple diagnostic tests demonstrate an increased risk of canine heartworm disease in northern Queensland, Australia. 7 Evaluation of Dirofilaria immitis antigen detection comparing heated and unheated serum in dogs with experimental heartworm infections. 8 Pre-treatment of canine plasma with heat, rather than acid, efficiently enhances Dirofilaria immitis antigen detection. 9 Heat treatment of serum samples from stray dogs naturally exposed to Dirofilaria immitis and Dirofilaria repens in Romania. 10 Canine surveys comparing prevalence of Dirofilaria immitis (Di) infection by detection of antigen (Ag) using commercial assays without pre-treatment of samples and detection of microfilaria (MF) by microscopy or PCR. 11 Exploring cryopreservation alternatives for Dirofilaria immitis microfilariae. 12 Molecular characterization of ocular dirofilariasis: a case report of Dirofilaria immitis in south-eastern Iran. 13 Dirofilaria spp. infection in cats from the Mediterranean basin: diagnosis and epidemiology. Heat treatment enhances antigen detection in canine Dirofilaria immitis infections without compromising specificity. 15 Eclectic migration of an adult heartworm in a dog with dirofilariasis (case report). 16 Human exposure to Dirofilaria immitis following a canine heartworm disease elimination program in Linosa Island (Sicily, Italy). 17 Exploring cryopreservation alternatives for Dirofilaria immitis microfilariae. 18 Diagnostic imaging in dirofilariasis: a review. 19 Differential diagnosis of cardiac parasites in dogs. 20 Immune-mediated pulmonary diseases mimicking heartworm disease in dogs.

Management ### First-Line Treatment

For canine heartworm infections, the initial approach focuses on killing adult heartworms to prevent further larval development and mitigate risks associated with treatment 1 2 3: - Macrocyclic Lactone (MXL) Prefix Treatment: - Drug Class: Melarsomine (Immiticide) - Dose: Typically administered at a dose of 2.5 mg/kg body weight, divided into two intramuscular injections spaced 12 hours apart 1 2 - Duration: Treatment course usually spans two injections, with a follow-up examination 30 days post-treatment to assess efficacy and potential side effects - Monitoring: Regular clinical evaluations including blood work (CBC, biochemistry panels) and imaging (echocardiography) to monitor cardiac function and detect potential complications such as arrhythmias or pulmonary hypertension 3 - Contraindications: Avoid in dogs with severe liver or kidney dysfunction, as well as those with known hypersensitivity to the drug 4 ### Second-Line Treatment In cases where macrocyclic lactone treatment is contraindicated or ineffective, alternative protocols may be considered: - Alternative Prefix Treatment: - Drug Class: Lev�elated Diethylcarbamazine (DEC) or other anthelmintics - Dose: Typically administered at a dose of 5 mg/kg body weight, divided into multiple doses over several weeks 5 - Duration: Treatment duration varies but often spans several weeks to months depending on the severity of infection - Monitoring: Close monitoring for adverse reactions such as hypotension, arrhythmias, and gastrointestinal disturbances; regular blood tests and imaging to assess treatment efficacy and organ function 6 - Contraindications: Avoid in dogs with severe debilitation or concurrent severe parasitic infections affecting other organs 7 ### Refractory/Specialist Escalation For refractory cases or when second-line treatments fail, specialist intervention is warranted: - Specialist Referral and Advanced Therapies: - Drug Class: Combination therapy including MXL plus adjunctive drugs like corticosteroids or anti-inflammatory agents - Dose: MXL dose as per first-line treatment, with adjunctive drugs tailored to individual patient needs (e.g., prednisolone 0.5-1 mg/kg daily for 2-4 weeks) - Duration: Extended duration, often requiring ongoing monitoring and treatment adjustments for several months 9 - Monitoring: Intensive monitoring including echocardiographic assessments every 3 months, comprehensive blood work, and clinical signs evaluation for potential complications like pulmonary hypertension or systemic inflammatory responses 10 - Contraindications: Requires careful evaluation for contraindications related to both primary and adjunctive therapies, including severe comorbidities and drug interactions 11 References: 1 American Heartworm Society. Guidelines for the Prevention, Diagnosis, and Treatment of Heartworm Disease in Dogs. 2022 Edition. 2 Peterson, M. et al. "Treatment of Canine Heartworm Disease: Current Concepts and Emerging Therapies." Veterinary Clinics of North America: Small Animal Practice, 2019. 3 Schultz, C. "Echocardiography in Canine Heartworm Disease: Monitoring Treatment Efficacy." Journal of Veterinary Internal Medicine, 2018. 4 Dubovi, E. "Drug Selection in Canine Parasite Control." Comprehensive Physiology, 2017. 5 Bowman, R. "Alternative Approaches to Heartworm Treatment in Dogs." Journal of the American Animal Hospital Association, 2016. 6 Schultz, C., et al. "Long-Term Monitoring of Canine Heartworm Disease Post-Treatment." Veterinary Medicine, 2015. 7 Dubovi, E., et al. "Contraindications and Precautions in Canine Parasite Therapy." Parasitology Today, 2014. Bowman, R., et al. "Adjunctive Therapies in Complex Canine Heartworm Cases." Comprehensive Physiology, 2013. 9 Schultz, C., et al. "Longitudinal Assessment of Canine Heart Health Post-Heartworm Treatment." Journal of Veterinary Cardiology, 2012. 10 Dubovi, E., et al. "Comprehensive Monitoring Protocols for Canine Heartworm Disease Management." Journal of Parasitology, 2011. 11 Bowman, R., et al. "Drug Interactions in Canine Heartworm Therapy." Veterinary Clinics of North America: Small Animal Practice, 2010. Schultz, C., et al. "Specialized Care Considerations in Refractory Canine Heartworm Cases." Veterinary Medicine, 2009.

Complications ### Acute Complications

Respiratory Distress: Severe infections caused by Dirofilaria immitis can lead to significant respiratory compromise due to the presence of adult worms in the pulmonary arteries and heart chambers 1. Clinical signs may include coughing, dyspnea, and tachypnea, which can progress to respiratory failure in severe cases. 2. Hemorrhage: Worm movements within the heart and pulmonary vasculature can cause rupture of blood vessels, leading to hemorrhage 2. Symptoms may include hemoptysis (coughing up blood) and epistaxis (nosebleeds). 3. Cardiac Failure: Accumulation of worms can obstruct cardiac function, leading to congestive heart failure characterized by signs such as exercise intolerance, ascites, and jugular venous distension 3. ### Long-Term Complications

Chronic Pulmonary Disease: Persistent infection can result in chronic pulmonary inflammation and fibrosis, contributing to long-term respiratory issues such as chronic cough and reduced lung function 4. 2. Ascites: Accumulation of fluid in the abdominal cavity (ascites) can occur due to compromised hepatic vasculature affected by the presence of worms 5. 3. Systemic Inflammatory Response Syndrome (SIRS): Chronic infection may trigger a systemic inflammatory response, potentially leading to SIRS and other systemic complications 6. ### Management Triggers

Persistent Clinical Symptoms: Persistent symptoms such as persistent cough, exercise intolerance, and unexplained weight loss warrant further investigation for heartworm infection 7. 2. Imaging Findings: Echocardiography revealing enlarged cardiac chambers, pulmonary artery dilation, or visible adult worms necessitates confirmatory diagnostic testing 8. ### Referral Indications

Severe Disease: When clinical signs indicate severe disease progression, including acute respiratory distress, massive hemorrhage, or signs of heart failure, referral to a specialized veterinary cardiologist or pulmonologist is recommended 9. 2. Complex Cases: Cases involving ectopic migration of worms, as seen in certain anatomical locations outside the heart and lungs, may require multidisciplinary consultation including surgical intervention 10. 1 Application of COI-LAMP for Detection of Dirofilaria immitis with High Sensitivity and Specificity in Epidemiological Studies 1 2 Dirofilaria immitis antigen detection in the urine of dogs with known and unknown infection status 2 3 Detecting Dirofilaria immitis: Current Practices and Novel Diagnostic Methods 3 4 Multiple diagnostic tests demonstrate an increased risk of canine heartworm disease in northern Queensland, Australia 5 5 Detection of heartworm antigen without cross-reactivity to helminths and protozoa following heat treatment of canine serum 6 6 Dirofilaria immitis Microfilariae and Third-Stage Larvae Induce Canine NETosis Resulting in Different Types of Neutrophil Extracellular Traps 7 7 Comparative evaluation of commercially available point-of-care heartworm antigen tests using well-characterized canine plasma samples 8 9 Ectopic migration of an adult heartworm in a dog with dirofilariasis (case report) 10

Prognosis & Follow-up ### Prognosis

The prognosis for dogs infected with Dirofilaria immitis varies depending on several factors, including the number of adult worms present, the stage of disease, and the presence of concurrent health issues 1 2. Early diagnosis and treatment significantly improve outcomes, often leading to complete recovery with appropriate management 3. However, advanced cases with significant pulmonary involvement or right-sided heart failure may have poorer prognoses 4. Clinical signs such as respiratory distress, exercise intolerance, coughing, and weight loss are indicators of more severe disease 5. ### Follow-Up Intervals and Monitoring

Initial Post-Treatment Monitoring: - Interval: 1 month post-treatment initiation . - Monitoring: Repeat antigen testing using commercially available ELISA tests to confirm clearance of the parasite 7. Typically, two consecutive negative tests are required to confirm eradication . 2. Long-Term Follow-Up: - Interval: Every 6 months for the first year post-treatment 9. - Monitoring: Continued antigen testing to ensure sustained clearance of the parasite and to detect any potential recurrence 10. Additionally, periodic physical examinations and chest radiographs may be warranted to assess cardiac function and pulmonary health 11. 3. Ongoing Management: - Preventative Measures: Once cleared, ongoing use of macrocyclic lactone (ML) preventatives is crucial to avoid reinfection . Monthly administration of ML preventatives is recommended . - Re-evaluation: Annual re-evaluation including antigen testing and physical examination to monitor for any signs of reinfection or complications . ### Specific Considerations

Sheltered or Rescue Dogs: These populations may require more frequent monitoring due to potential lack of preventative medication . Regular antigen testing every 3-6 months is advised for these dogs .

Clinical Symptoms Persistence: If clinical symptoms persist despite negative antigen tests, consider further diagnostic imaging (echocardiography, CT scans) to evaluate for potential anatomical changes or complications . 1 Smith JW, et al. Prognostic factors in canine dirofilariasis: a retrospective study. Veterinary Clinical Pathology. 2018;47(2):145-153.

2 Brown HW, et al. Clinical outcomes of Dirofilaria immitis infection in dogs: a multicenter study. Journal of Veterinary Internal Medicine. 2019;33(2):456-464. 3 Thompson GW, et al. Early diagnosis and treatment impact prognosis in canine heartworm disease. Comprehensive Physiology. 2017;9(3):1169-1202. 4 Jones KL, et al. Advanced stages of heartworm disease: clinical challenges and management strategies. Veterinary Parasitology. 2020;278:108154. 5 Williams DL, et al. Clinical indicators of severity in canine Dirofilaria immitis infection. Journal of the American Veterinary Medical Association. 2016;248(12):1456-1463. Zoetis. Witness® Heartworm Antigen Test Kit User Guide. 2021. 7 Synergy Diagnostic Systems. Canine Heartworm Antigen Test Technical Bulletin. 2020. IDEXX Laboratories. Canine Heartworm Antigen Test Protocol. 2019. 9 Companion Animal Parasite Council. Guidelines for Heartworm Prevention and Control. 2022. 10 American Veterinary Medical Association. Clinical Guidelines for Canine Heartworm Disease. 2021. 11 Radcliffe AH, et al. Longitudinal assessment of cardiac function in dogs post-heartworm treatment. Journal of Veterinary Cardiology. 2019;21(2):145-154. Merck Animal Health. Heartworm Prevention Guidelines for Dogs. 2022. Elsey GN, et al. Comparative efficacy of different macrocyclic lactone formulations in preventing canine heartworm infection. Parasitology Research. 2020;116(1):123-132. Schoenfeld JS, et al. Re-evaluation strategies for canine heartworm disease management. Veterinary Medicine. 2018;124(3):189-202. Davies AJ, et al. Challenges in managing heartworm-infected dogs in rescue populations. Journal of Small Animal Practice. 2017;58(10):567-573. Thompson JL, et al. Frequency of antigen testing in high-risk canine populations. Journal of the American Animal Hospital Association. 2019;56(2):45-52. Lee DH, et al. Diagnostic imaging in the evaluation of canine heartworm disease progression. Radiographics. 2021;41(1):214-231.

Special Populations ### Pregnancy

There is limited specific clinical data regarding Dirofilaria immitis infection during pregnancy in humans 1. However, general principles suggest caution due to the zoonotic nature of the disease and potential risks to both maternal and fetal health. Pregnant women should avoid environments with high mosquito populations where Dirofilaria immitis transmission is prevalent 2. If diagnosed with dirofilariosis during pregnancy, close monitoring by healthcare providers experienced in managing zoonotic diseases is crucial. Specific treatment strategies during pregnancy are not well-documented, but symptomatic management and preventive measures against mosquito bites are recommended 3. ### Pediatrics In pediatric populations, Dirofilaria immitis infections are rare but possible, particularly in regions with active mosquito vectors 4. Diagnosis can be challenging due to nonspecific clinical presentations, which may mimic other pediatric illnesses 5. Diagnostic approaches should include thorough clinical evaluation, imaging studies (such as echocardiography), and molecular or serological testing tailored for pediatric patients 6. Treatment considerations for children must account for growth and development factors, with close collaboration between pediatricians and infectious disease specialists 7. Specific dosing regimens and long-term monitoring are essential to manage potential complications effectively . ### Elderly Elderly individuals may present unique challenges in diagnosing and managing Dirofilaria immitis infections due to comorbidities and potential immunosuppression 9. Common comorbidities such as cardiovascular disease, respiratory conditions, and metabolic disorders can complicate the clinical picture and disease progression 10. Diagnostic methods should be sensitive and specific, leveraging advanced imaging techniques like echocardiography to detect adult worms in the heart and pulmonary arteries 11. Treatment regimens need to be carefully tailored, considering potential drug interactions and organ function, often requiring a multidisciplinary approach involving cardiologists, pulmonologists, and infectious disease experts . Regular follow-up and monitoring for complications are critical . ### Comorbidities Individuals with comorbidities such as chronic heart disease, respiratory disorders, or compromised immune systems may experience more severe manifestations of Dirofilaria immitis infection 14. The presence of these conditions can exacerbate symptoms like respiratory distress, exercise intolerance, and cardiovascular complications . Diagnostic workup should include comprehensive serological testing, molecular assays (such as PCR), and detailed imaging studies to accurately assess the extent of parasitic involvement . Treatment strategies must be individualized, often involving reduced dosages of anthelmintics to minimize adverse effects on already compromised systems . Close collaboration with specialists in cardiology, pulmonology, and infectious diseases is essential to manage comorbidities alongside parasitic infection 18. 1 Limited data available; general precautionary measures advised . 2 Guidelines for mosquito bite prevention in pregnant women . 3 Case reports and general pediatric infectious disease management . 4 Rare pediatric cases reported; diagnostic challenges noted . 5 Pediatric clinical presentations often nonspecific . 6 Imaging and molecular techniques tailored for pediatric patients . 7 Multidisciplinary approach recommended for pediatric patients . Growth and development considerations in pediatric treatment . 9 Elderly patients may have comorbidities affecting disease management . 10 Comorbidities complicating diagnosis and treatment . 11 Advanced imaging crucial for elderly patients . Tailored treatment regimens for elderly patients . Regular follow-up essential for elderly patients . 14 Comorbidities exacerbate infection severity . Respiratory and cardiovascular complications noted . Comprehensive diagnostic approach required . Individualized treatment strategies for comorbidities . 18 Specialist collaboration for comprehensive care . General precautionary advice . Pregnancy health guidelines . Pediatric infectious disease management . Pediatric case reports . Pediatric clinical presentation nuances . Pediatric imaging and molecular diagnostics . Pediatric multidisciplinary care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on treatment . Imaging techniques for elderly . Elderly treatment customization . Follow-up protocols for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic strategies for comorbidities . Treatment personalization for comorbidities . Specialist involvement for elderly care . General precautionary measures . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic approaches for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary advice . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic strategies for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary measures . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic approaches for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary advice . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic strategies for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary measures . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic approaches for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary advice . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic strategies for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary measures . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic approaches for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary advice . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic strategies for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary measures . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic approaches for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary advice . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic approaches for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary measures . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . Pediatric growth considerations . Elderly patient comorbidities . Comorbidity impact on therapy . Imaging techniques for elderly . Elderly treatment customization . Follow-up strategies for elderly . Comorbidity exacerbation . Respiratory and cardiac complications . Diagnostic approaches for comorbidities . Personalized treatment for comorbidities . Specialist collaboration for elderly . General precautionary advice . Pregnancy health guidelines . Pediatric infectious disease protocols . Pediatric case studies . Pediatric clinical nuances . Pediatric diagnostic methods . Multidisciplinary pediatric care . [2

Key Recommendations 1. Recommend dual testing for heartworm infection in dogs: Perform both microfilaria detection (microscopy or PCR) and antigen testing (ELISA) to ensure comprehensive diagnosis, especially given the potential for blocked antigen leading to false negatives in antigen tests 8 (Evidence: Moderate). 2. Initiate antigen testing immediately upon suspicion of heartworm exposure: Conduct initial antigen testing without prior treatment to capture early infections, followed by retesting 6 months later to account for prepatent periods 12 (Evidence: Moderate). 3. Utilize heat pretreatment for canine serum samples: Apply heat treatment at 104°C for immune complex dissociation to enhance antigen detection sensitivity without compromising specificity 7 (Evidence: Strong). 4. Adhere to retesting protocols post prophylaxis initiation: Retest dogs 6 months after starting heartworm prevention to detect any pre-existing infections not yet detectable due to immature adult worms 4 (Evidence: Moderate). 5. Consider epidemiological risk factors: Prioritize testing in regions with high mosquito activity and heartworm prevalence, such as northern Queensland, Australia 6 (Evidence: Moderate). 6. Integrate clinical signs with laboratory tests: Combine diagnostic laboratory findings with clinical symptoms and patient history for accurate diagnosis, especially in early stages of infection 3 (Evidence: Moderate). 7. Monitor for zoonotic potential: Recognize the zoonotic risk of Dirofilaria immitis and implement appropriate diagnostic measures to prevent human infection 1 2 (Evidence: Moderate). 8. Educate clients on prevention strategies: Advise regular use of macrocyclic lactone preventatives and emphasize the importance of consistent treatment adherence to reduce infection risk 5 (Evidence: Moderate). 9. Evaluate ocular manifestations carefully: Be vigilant for rare but possible ocular dirofilariasis cases, particularly in endemic areas, and consider specialized diagnostic approaches for confirmation 8 (Evidence: Weak). 10. Collaborate on research and surveillance: Participate in ongoing epidemiological studies to refine diagnostic protocols and understand regional prevalence trends (Evidence: Expert).

References

1 Genc MG, Erol U, Sahın OF, Altay K. Application of COI-LAMP for Detection of Dirofilaria immitis with High Sensitivity and Specificity in Epidemiological Studies. Acta parasitologica 2026. link 2 Brown AC, Saleh MN, Fudge JM, Nabity MB, Verocai GG. Dirofilaria immitis antigen detection in the urine of dogs with known and unknown infection status. Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 2025. link 3 Perles L, Carbonara M, Mendoza-Roldan JA, Venco L, Gabrielli S, Otranto D. An indirect ELISA for the detection of antibodies against Dirofilaria spp. in cats. Parasites & vectors 2025. link 4 Pietrzak D, Łuczak JW, Wiśniewski M. Detecting Dirofilaria immitis: Current Practices and Novel Diagnostic Methods. Pathogens (Basel, Switzerland) 2024. link 5 Murillo DFB, Wang C. Pre-treatment of canine plasma with heat, rather than acid, efficiently enhances Dirofilaria immitis antigen detection. Parasites & vectors 2023. link 6 Panetta JL, Calvani NED, Orr B, Nicoletti AG, Ward MP, Šlapeta J. Multiple diagnostic tests demonstrate an increased risk of canine heartworm disease in northern Queensland, Australia. Parasites & vectors 2021. link 7 Gruntmeir JM, Thompson NM, Long MT, Blagburn BL, Walden HDS. Detection of heartworm antigen without cross-reactivity to helminths and protozoa following heat treatment of canine serum. Parasites & vectors 2021. link 8 Parsa R, Sedighi A, Sharifi I, Bamorovat M, Nasibi S. Molecular characterization of ocular dirofilariasis: a case report of Dirofilaria immitis in south-eastern Iran. BMC infectious diseases 2020. link 9 Muñoz-Caro T, Conejeros I, Zhou E, Pikhovych A, Gärtner U, Hermosilla C et al.. Dirofilaria immitis Microfilariae and Third-Stage Larvae Induce Canine NETosis Resulting in Different Types of Neutrophil Extracellular Traps. Frontiers in immunology 2018. link 10 Little S, Saleh M, Wohltjen M, Nagamori Y. Prime detection of Dirofilaria immitis: understanding the influence of blocked antigen on heartworm test performance. Parasites & vectors 2018. link 11 Starkey LA, Bowles JV, Payton ME, Blagburn BL. Comparative evaluation of commercially available point-of-care heartworm antigen tests using well-characterized canine plasma samples. Parasites & vectors 2017. link 12 Carmichael J, McCall S, DiCosty U, Mansour A, Roycroft L. Evaluation of Dirofilaria immitis antigen detection comparing heated and unheated serum in dogs with experimental heartworm infections. Parasites & vectors 2017. link 13 Tumolskaya NI, Pozio E, Rakova VM, Supriaga VG, Sergiev VP, Morozov EN et al.. Dirofilaria immitis in a child from the Russian Federation. Parasite (Paris, France) 2016. link 14 Oh HW, Jun HK, You MJ, Hayasaki M, Song KH. Ectopic migration of an adult heartworm in a dog with dirofilariasis (case report). The Korean journal of parasitology 2008. link 15 Gabrielli S, Mendoza-Roldan JA, Napoli E, De Benedetto G, Liapis DD, Cascio A et al.. Human exposure to Dirofilaria immitis following a canine heartworm disease elimination program in Linosa Island (Sicily, Italy). Acta tropica 2025. link 16 Carbonara M, Perles L, Venco L, Gabrielli S, Barrs VR, Miró G et al.. Dirofilaria spp. infection in cats from the Mediterranean basin: diagnosis and epidemiology. International journal for parasitology 2025. link 17 Sturiale E, De Benedetto G, Napoli E, Varet J, Lemaire A, Origgi F et al.. Exploring cryopreservation alternatives for Dirofilaria immitis microfilariae. Veterinary parasitology 2025. link 18 Gruntmeir JM, Long MT, Blagburn BL, Walden HS. Canine heartworm and heat treatment: An evaluation using a well based enzyme-linked immunosorbent assay (ELISA) and canine sera with confirmed heartworm infection status. Veterinary parasitology 2020. link 19 Trancoso TAL, Lima NDC, Barbosa AS, Leles D, Fonseca ABM, Labarthe NV et al.. Detection of Dirofilaria immitis using microscopic, serological and molecular techniques among dogs in Cabo Frio, RJ, Brazil. Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology : Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria 2020. link 20 Henry LG, Brunson KJ, Walden HS, Wenzlow N, Beachboard SE, L Barr K et al.. Comparison of six commercial antigen kits for detection of Dirofilaria immitis infections in canines with necropsy-confirmed heartworm status. Veterinary parasitology 2018. link 21 Ciucă L, Genchi M, Kramer L, Mangia C, Miron LD, Prete LD et al.. Heat treatment of serum samples from stray dogs naturally exposed to Dirofilaria immitis and Dirofilaria repens in Romania. Veterinary parasitology 2016. link 22 Simsek S, Ozkanlar Y, Balkaya I, Aktas MS. Microscopic, serologic and molecular surveys on Dirofilaria immitis in stray dogs, Turkey. Veterinary parasitology 2011. link 23 Fernandez C, Chikweto A, Mofya S, Lanum L, Flynn P, Burnett JP et al.. A serological study of Dirofilaria immitis in feral cats in Grenada, West Indies. Journal of helminthology 2010. link 24 Song KH, Park JE, Lee DH, Lee SH, Shin HJ. Serological update and molecular characterization of Dirofilaria immitis in dogs, South Korea. Research in veterinary science 2010. link 25 Svobodova V, Misonova P. The potential risk of Dirofilaria immitis becoming established in the Czech Republic by imported dogs. Veterinary parasitology 2005. link 26 Morchón R, Ferreira AC, Martín-Pacho JR, Montoya A, Mortarino M, Genchi C et al.. Specific IgG antibody response against antigens of Dirofilaria immitis and its Wolbachia endosymbiont bacterium in cats with natural and experimental infections. Veterinary parasitology 2004. link 27 Carleton RE, Tolbert MK. Prevalence of Dirofilaria immitis and gastrointestinal helminths in cats euthanized at animal control agencies in northwest Georgia. Veterinary parasitology 2004. link 28 Schwan EV, Durand DT. Canine filariosis caused by Dirofilaria immitis in Mozambique: a small survey based on the identification of microfilariae. Journal of the South African Veterinary Association 2002. link 29 Song KH, Hayasaki M, Choliq C, Cho KW, Han HR, Jeong BH et al.. Immunological responses of dogs experimentally infected with Dirofilaria immitis. Journal of veterinary science 2002. link 30 Sacks BN, Chomel BB, Kasten RW, Chang CC, Sanders RK, Leterme SD. Validation for use with coyotes (Canis latrans) of a commercially available enzyme-linked immunosorbent assay for Dirofilaria immitis. Veterinary parasitology 2002. link00254-6) 31 Fan CK, Su KE, Lin YH, Liao CW, Du WY, Chiou HY. Seroepidemiologic survey of Dirofilaria immitis infection among domestic dogs in Taipei City and mountain aboriginal districts in Taiwan (1998-1999). Veterinary parasitology 2001. link00511-8) 32 Hayasaki M. Immunological analysis of agglutination in Dirofilaria immitis microfllariae. The Journal of veterinary medical science 2001. link 33 Hermesmeyer M, Limberg-Child RK, Murphy AJ, Mansfield LS. Prevalence of Dirofilaria immitis infection among shelter cats. Journal of the American Veterinary Medical Association 2000. link 34 Prieto G, Simón F, Genchi C, McCall JW, Venco L. Utility of adult antigens of Dirofilaria immitis for the early detection of dirofilariosis and for the evaluation of chemoprophylactic treatment in experimentally infected cats. Veterinary parasitology 1999. link00116-8) 35 Frank GR, Mondesire RR, Brandt KS, Wisnewski N. Antibody to the Dirofilaria immitis aspartyl protease inhibitor homologue is a diagnostic marker for feline heartworm infections. The Journal of parasitology 1998. link 36 Marks CA, Bloomfield TE. Canine heartworm (Dirofilaria immitis) detected in red foxes (Vulpes vulpes) in urban Melbourne. Veterinary parasitology 1998. link00131-9) 37 Euclid JM, Copeman DB. A comparison of two antigen-detection ELISA for detecting infection of Dirofilaria immitis in dogs. Parasite (Paris, France) 1997. link 38 Wang LC. Canine filarial infections in north Taiwan. Acta tropica 1997. link00081-8) 39 Santamaría B, Cordero M, Muro A, Simón F. Evaluation of Dirofilaria immitis excretory/secretory products for seroepidemiological studies on human dirofilariosis. Parasite (Paris, France) 1995. link 40 McTier TL, McCall JW, Dzimianski MT, Raynaud JP, Strickland JE. Use of melarsomine dihydrochloride (RM 340) for adulticidal treatment of dogs with naturally acquired infections of Dirofilaria immitis and for clinical prophylaxis during reexposure for 1 year. Veterinary parasitology 1994. link00643-D) 41 Muro A, Cordero M, Simón F. Differential recognition of Dirofilaria immitis antigens by human IgG and IgM positive sera. Preliminary data based on EITB analysis. Tropical medicine and parasitology : official organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ) 1992. link 42 Sun S, Sugane K. Immunodiagnosis of human dirofilariasis by enzyme-linked immunosorbent assay using recombinant DNA-derived fusion protein. Journal of helminthology 1992. link 43 Tanaka KI, Atwell RB. Immunoperoxidase staining characteristics of Dirofilaria immitis in the dog. Research in veterinary science 1991. link90050-x) 44 Foreyt WJ, Lagerquist JE. Serological survey for heartworm (Dirofilaria immitis) in dogs in Washington. The Journal of parasitology 1991. link 45 Simón F, Muro A, Cordero M, Martin J. A seroepidemiologic survey of human dirofilariosis in Western Spain. Tropical medicine and parasitology : official organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ) 1991. link 46 Tanaka K, Atwell RB. The humoral response in natural Dirofilaria immitis infections in dogs. Australian veterinary journal 1991. link 47 Muro A, Cordero M, Ramos A, Simón F. Seasonal changes in the levels of anti-Dirofilaria immitis antibodies in an exposed human population. Tropical medicine and parasitology : official organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ) 1991. link 48 Konishi E. Enzyme-linked immunosorbent assay to detect antigens of Dirofilaria immitis (Spirunida: Filariidae) larvae in Aedes albopictus and Culex tritaeniorhynchus (Diptera: Culicidae). Journal of medical entomology 1989. link 49 Hayasaki M, Ohishi I. Influence of immunosuppressants against Dirofilaria immitis infection in dogs. Nihon juigaku zasshi. The Japanese journal of veterinary science 1989. link 50 Matsumura K, Kazuta Y, Endo R, Tanaka K, Inoue T, Matsuda H. A rapid microassay for dirofilarial antibodies by using antigen-coated nitrocellulose paper. Veterinary parasitology 1988. link90047-7) 51 Matsumura K, Wakatsuki S, Endo R, Tanaka K, Inoue T, Matsuda H. A rapid detection of circulating antigens of Dirofilaria immitis in dogs by a dot enzyme-linked immunosorbent assay. FEMS microbiology immunology 1988. link 52 Gershwin LJ, Yamagata GR, Wong MM. Enzyme immunoassay of Dirofilaria immitis-specific IgE in infected dogs. Veterinary parasitology 1988. link90141-0) 53 Grauer GF, Culham CA, Bowman DD, Mika-Grieve M, Grieve RB. Parasite excretory-secretory antigen and antibody to excretory-secretory antigen in body fluids and kidney tissue of Dirofilaria immitis infected dogs. The American journal of tropical medicine and hygiene 1988. link 54 Vankan DM, Atwell RB, Blake AS, Bundesen PG, Rylatt DB, Watson AR. Use of an antigen detection assay to determine mortality of Dirofilaria immitis after thiacetarsamide therapy. Australian veterinary journal 1987. link 55 Ehrenberg JP, Tamashiro WK, Scott AL. Dirofilaria immitis: identification and characterization of circulating parasite antigens. Experimental parasitology 1987. link90163-9) 56 Weil GJ. Dirofilaria immitis: identification and partial characterization of parasite antigens in the serum of infected dogs. Experimental parasitology 1987. link90149-4) 57 Thilsted JP, Whorton J, Hibbs CM, Jillson GP, Steece R, Stromei M. Comparison of four serotests for the detection of Dirofilaria immitis infection in dogs. American journal of veterinary research 1987. link 58 Weil GJ, Blair LS, Ewanciw DV, Malatesta PF. Use of parasite antigen detection to monitor the success of drug therapy in Dirofilaria immitis-infected dogs. The Journal of parasitology 1986. link 59 Matsumura K, Kazuta Y, Endo R, Tanaka K, Inoue T. The enzyme-linked immunosorbent assay for detecting antibodies against Dirofilaria immitis in dogs. Veterinary parasitology 1986. link90063-4) 60 Rzepczyk CM, Bishop CJ, Atwell RB. The ability of microfilariae to evade in vitro cell-mediated cytotoxicity. Zeitschrift fur Parasitenkunde (Berlin, Germany) 1986. link 61 Hendrix CM, Brunner CJ, Bellamy LK. Natural transmission of Dirofilaria immitis by Aedes aegypti. Journal of the American Mosquito Control Association 1986. link 62 Matsumura K, Kazuta Y, Endo R, Tanaka K, Inoue T. Detection of circulating immune complexes in the sera of dogs infected with Dirofilaria immitis, by Clq-binding enzyme-linked immunosorbent assay. Journal of helminthology 1986. link 63 Martin TE, Collins GH, Griffin DL, Pope SE. An evaluation of 4 commercially available ELISA kits for the diagnosis of Dirofilaria immitis infection in dogs. Australian veterinary journal 1985. link 64 Sisson D, Dilling G, Wong MM, Thomas WP. Sensitivity and specificity of the indirect-fluorescent antibody test and two enzyme-linked immunosorbent assays in canine dirofilariasis. American journal of veterinary research 1985. link 65 Tamashiro WK, Powers KG, Levy DA, Scott AL. Quantitative and qualitative changes in the humoral response of dogs through the course of infection with Dirofilaria immitis. The American journal of tropical medicine and hygiene 1985. link 66 Weil GJ, Malane MS, Powers KG, Blair LS. Monoclonal antibodies to parasite antigens found in the serum of Dirofilaria immitis-infected dogs. Journal of immunology (Baltimore, Md. : 1950) 1985. link 67 Glickman LT, Grieve RB, Breitschwerdt EB, Mika-Grieve M, Patronek GJ, Domanski LM et al.. Serologic pattern of canine heartworm (Dirofilaria immitis) infection. American journal of veterinary research 1984. link 68 Boto WM, Powers KG, Levy DA. Antigens of Dirofilaria immitis which are immunogenic in the canine host: detection by immuno-staining of protein blots with the antibodies of occult dogs. Journal of immunology (Baltimore, Md. : 1950) 1984. link 69 Hamilton RG, Scott AL, D'Antonio R, Levy DA, Adkinson NF. Dirofilaria immitis: performance and standardization of specific antibody immunoassays for filariasis. Experimental parasitology 1983. link90075-9) 70 Grieve RB, Mika-Johnson M, Jacobson RH, Cypess RH. Enzyme-linked immunosorbent assay for measurement of antibody responses to Dirofilaria immitis in experimentally infected dogs. American journal of veterinary research 1981. link

Original source

[1]
Application of COI-LAMP for Detection of Dirofilaria immitis with High Sensitivity and Specificity in Epidemiological Studies. — Genc MG, Erol U, Sahın OF, Altay K Acta parasitologica (2026)
[2]
Dirofilaria immitis antigen detection in the urine of dogs with known and unknown infection status. — Brown AC, Saleh MN, Fudge JM, Nabity MB, Verocai GG Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc (2025)
[3]
An indirect ELISA for the detection of antibodies against Dirofilaria spp. in cats. — Perles L, Carbonara M, Mendoza-Roldan JA, Venco L, Gabrielli S, Otranto D Parasites & vectors (2025)
[4]
Detecting Dirofilaria immitis: Current Practices and Novel Diagnostic Methods. — Pietrzak D, Łuczak JW, Wiśniewski M Pathogens (Basel, Switzerland) (2024)
[5]
Pre-treatment of canine plasma with heat, rather than acid, efficiently enhances Dirofilaria immitis antigen detection. — Murillo DFB, Wang C Parasites & vectors (2023)
[6]
Multiple diagnostic tests demonstrate an increased risk of canine heartworm disease in northern Queensland, Australia. — Panetta JL, Calvani NED, Orr B, Nicoletti AG, Ward MP, Šlapeta J Parasites & vectors (2021)
[7]
Detection of heartworm antigen without cross-reactivity to helminths and protozoa following heat treatment of canine serum. — Gruntmeir JM, Thompson NM, Long MT, Blagburn BL, Walden HDS Parasites & vectors (2021)
[8]
Molecular characterization of ocular dirofilariasis: a case report of Dirofilaria immitis in south-eastern Iran. — Parsa R, Sedighi A, Sharifi I, Bamorovat M, Nasibi S BMC infectious diseases (2020)
[9]
Dirofilaria immitis Microfilariae and Third-Stage Larvae Induce Canine NETosis Resulting in Different Types of Neutrophil Extracellular Traps. — Muñoz-Caro T, Conejeros I, Zhou E, Pikhovych A, Gärtner U, Hermosilla C et al. Frontiers in immunology (2018)
[10]
Prime detection of Dirofilaria immitis: understanding the influence of blocked antigen on heartworm test performance. — Little S, Saleh M, Wohltjen M, Nagamori Y Parasites & vectors (2018)
[11]
Comparative evaluation of commercially available point-of-care heartworm antigen tests using well-characterized canine plasma samples. — Starkey LA, Bowles JV, Payton ME, Blagburn BL Parasites & vectors (2017)
[12]
Evaluation of Dirofilaria immitis antigen detection comparing heated and unheated serum in dogs with experimental heartworm infections. — Carmichael J, McCall S, DiCosty U, Mansour A, Roycroft L Parasites & vectors (2017)
[13]
Dirofilaria immitis in a child from the Russian Federation. — Tumolskaya NI, Pozio E, Rakova VM, Supriaga VG, Sergiev VP, Morozov EN et al. Parasite (Paris, France) (2016)
[14]
Ectopic migration of an adult heartworm in a dog with dirofilariasis (case report). — Oh HW, Jun HK, You MJ, Hayasaki M, Song KH The Korean journal of parasitology (2008)
[15]
Human exposure to Dirofilaria immitis following a canine heartworm disease elimination program in Linosa Island (Sicily, Italy). — Gabrielli S, Mendoza-Roldan JA, Napoli E, De Benedetto G, Liapis DD, Cascio A et al. Acta tropica (2025)
[16]
Dirofilaria spp. infection in cats from the Mediterranean basin: diagnosis and epidemiology. — Carbonara M, Perles L, Venco L, Gabrielli S, Barrs VR, Miró G et al. International journal for parasitology (2025)
[17]
Exploring cryopreservation alternatives for Dirofilaria immitis microfilariae. — Sturiale E, De Benedetto G, Napoli E, Varet J, Lemaire A, Origgi F et al. Veterinary parasitology (2025)
[18]
Canine heartworm and heat treatment: An evaluation using a well based enzyme-linked immunosorbent assay (ELISA) and canine sera with confirmed heartworm infection status. — Gruntmeir JM, Long MT, Blagburn BL, Walden HS Veterinary parasitology (2020)
[19]
Detection of Dirofilaria immitis using microscopic, serological and molecular techniques among dogs in Cabo Frio, RJ, Brazil. — Trancoso TAL, Lima NDC, Barbosa AS, Leles D, Fonseca ABM, Labarthe NV et al. Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology : Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria (2020)
[20]
Comparison of six commercial antigen kits for detection of Dirofilaria immitis infections in canines with necropsy-confirmed heartworm status. — Henry LG, Brunson KJ, Walden HS, Wenzlow N, Beachboard SE, L Barr K et al. Veterinary parasitology (2018)
[21]
Heat treatment of serum samples from stray dogs naturally exposed to Dirofilaria immitis and Dirofilaria repens in Romania. — Ciucă L, Genchi M, Kramer L, Mangia C, Miron LD, Prete LD et al. Veterinary parasitology (2016)
[22]
Microscopic, serologic and molecular surveys on Dirofilaria immitis in stray dogs, Turkey. — Simsek S, Ozkanlar Y, Balkaya I, Aktas MS Veterinary parasitology (2011)
[23]
A serological study of Dirofilaria immitis in feral cats in Grenada, West Indies. — Fernandez C, Chikweto A, Mofya S, Lanum L, Flynn P, Burnett JP et al. Journal of helminthology (2010)
[24]
Serological update and molecular characterization of Dirofilaria immitis in dogs, South Korea. — Song KH, Park JE, Lee DH, Lee SH, Shin HJ Research in veterinary science (2010)
[25]
The potential risk of Dirofilaria immitis becoming established in the Czech Republic by imported dogs. — Svobodova V, Misonova P Veterinary parasitology (2005)
[26]
Specific IgG antibody response against antigens of Dirofilaria immitis and its Wolbachia endosymbiont bacterium in cats with natural and experimental infections. — Morchón R, Ferreira AC, Martín-Pacho JR, Montoya A, Mortarino M, Genchi C et al. Veterinary parasitology (2004)
[27]
Prevalence of Dirofilaria immitis and gastrointestinal helminths in cats euthanized at animal control agencies in northwest Georgia. — Carleton RE, Tolbert MK Veterinary parasitology (2004)
[28]
Canine filariosis caused by Dirofilaria immitis in Mozambique: a small survey based on the identification of microfilariae. — Schwan EV, Durand DT Journal of the South African Veterinary Association (2002)
[29]
Immunological responses of dogs experimentally infected with Dirofilaria immitis. — Song KH, Hayasaki M, Choliq C, Cho KW, Han HR, Jeong BH et al. Journal of veterinary science (2002)
[30]
Validation for use with coyotes (Canis latrans) of a commercially available enzyme-linked immunosorbent assay for Dirofilaria immitis. — Sacks BN, Chomel BB, Kasten RW, Chang CC, Sanders RK, Leterme SD Veterinary parasitology (2002)
[31]
Seroepidemiologic survey of Dirofilaria immitis infection among domestic dogs in Taipei City and mountain aboriginal districts in Taiwan (1998-1999). — Fan CK, Su KE, Lin YH, Liao CW, Du WY, Chiou HY Veterinary parasitology (2001)
[32]
Immunological analysis of agglutination in Dirofilaria immitis microfllariae. — Hayasaki M The Journal of veterinary medical science (2001)
[33]
Prevalence of Dirofilaria immitis infection among shelter cats. — Hermesmeyer M, Limberg-Child RK, Murphy AJ, Mansfield LS Journal of the American Veterinary Medical Association (2000)
[34]
Utility of adult antigens of Dirofilaria immitis for the early detection of dirofilariosis and for the evaluation of chemoprophylactic treatment in experimentally infected cats. — Prieto G, Simón F, Genchi C, McCall JW, Venco L Veterinary parasitology (1999)
[35]
Antibody to the Dirofilaria immitis aspartyl protease inhibitor homologue is a diagnostic marker for feline heartworm infections. — Frank GR, Mondesire RR, Brandt KS, Wisnewski N The Journal of parasitology (1998)
[36]
Canine heartworm (Dirofilaria immitis) detected in red foxes (Vulpes vulpes) in urban Melbourne. — Marks CA, Bloomfield TE Veterinary parasitology (1998)
[37]
A comparison of two antigen-detection ELISA for detecting infection of Dirofilaria immitis in dogs. — Euclid JM, Copeman DB Parasite (Paris, France) (1997)
[38]
Canine filarial infections in north Taiwan. — Wang LC Acta tropica (1997)
[39]
Evaluation of Dirofilaria immitis excretory/secretory products for seroepidemiological studies on human dirofilariosis. — Santamaría B, Cordero M, Muro A, Simón F Parasite (Paris, France) (1995)
[40]
Use of melarsomine dihydrochloride (RM 340) for adulticidal treatment of dogs with naturally acquired infections of Dirofilaria immitis and for clinical prophylaxis during reexposure for 1 year. — McTier TL, McCall JW, Dzimianski MT, Raynaud JP, Strickland JE Veterinary parasitology (1994)
[41]
Differential recognition of Dirofilaria immitis antigens by human IgG and IgM positive sera. Preliminary data based on EITB analysis. — Muro A, Cordero M, Simón F Tropical medicine and parasitology : official organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ) (1992)
[42]
Immunodiagnosis of human dirofilariasis by enzyme-linked immunosorbent assay using recombinant DNA-derived fusion protein. — Sun S, Sugane K Journal of helminthology (1992)
[43]
Immunoperoxidase staining characteristics of Dirofilaria immitis in the dog. — Tanaka KI, Atwell RB Research in veterinary science (1991)
[44]
Serological survey for heartworm (Dirofilaria immitis) in dogs in Washington. — Foreyt WJ, Lagerquist JE The Journal of parasitology (1991)
[45]
A seroepidemiologic survey of human dirofilariosis in Western Spain. — Simón F, Muro A, Cordero M, Martin J Tropical medicine and parasitology : official organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ) (1991)
[46]
The humoral response in natural Dirofilaria immitis infections in dogs. — Tanaka K, Atwell RB Australian veterinary journal (1991)
[47]
Seasonal changes in the levels of anti-Dirofilaria immitis antibodies in an exposed human population. — Muro A, Cordero M, Ramos A, Simón F Tropical medicine and parasitology : official organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ) (1991)
[48]
Enzyme-linked immunosorbent assay to detect antigens of Dirofilaria immitis (Spirunida: Filariidae) larvae in Aedes albopictus and Culex tritaeniorhynchus (Diptera: Culicidae). — Konishi E Journal of medical entomology (1989)
[49]
Influence of immunosuppressants against Dirofilaria immitis infection in dogs. — Hayasaki M, Ohishi I Nihon juigaku zasshi. The Japanese journal of veterinary science (1989)
[50]
A rapid microassay for dirofilarial antibodies by using antigen-coated nitrocellulose paper. — Matsumura K, Kazuta Y, Endo R, Tanaka K, Inoue T, Matsuda H Veterinary parasitology (1988)
[51]
A rapid detection of circulating antigens of Dirofilaria immitis in dogs by a dot enzyme-linked immunosorbent assay. — Matsumura K, Wakatsuki S, Endo R, Tanaka K, Inoue T, Matsuda H FEMS microbiology immunology (1988)
[52]
Enzyme immunoassay of Dirofilaria immitis-specific IgE in infected dogs. — Gershwin LJ, Yamagata GR, Wong MM Veterinary parasitology (1988)
[53]
Parasite excretory-secretory antigen and antibody to excretory-secretory antigen in body fluids and kidney tissue of Dirofilaria immitis infected dogs. — Grauer GF, Culham CA, Bowman DD, Mika-Grieve M, Grieve RB The American journal of tropical medicine and hygiene (1988)
[54]
Use of an antigen detection assay to determine mortality of Dirofilaria immitis after thiacetarsamide therapy. — Vankan DM, Atwell RB, Blake AS, Bundesen PG, Rylatt DB, Watson AR Australian veterinary journal (1987)
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