American Heartworm Society Research Articles

Comparative performance analysis of different microfilaria testing methods for Dirofilaria immitis in canine blood

Abstract Background Microfilaria (MF) testing is an essential part of canine heartworm diagnostics, and it is recommended by the American Heartworm Society that a MF test be performed in tandem with antigen testing on every dog, every year, regardless of prevention status or history. There are a variety of methods that can be used to detect MF in canine whole blood; however, these methods widely vary in their sensitivities as well as practical factors, including time investment and cost. Additionally, some MF tests offer the advantage of being quantitative or allowing for morphological or molecular species identification, while other tests should only be used qualitatively. Methods The purpose of this study is to evaluate the quantitative and qualitative performance of MF tests, including the 20 μL count, wet mount, 9 μL and 40 μL hematocrit tubes, thin smear, thick smear, modified Knott test (MKT), and conventional polymerase chain reaction (PCR). Results Qualitatively, there was little difference in the performance of the 20 μL count, wet mount, MKT, and PCR. The MKT and PCR are the optimal MF tests, as these perform most reliably for detecting positives even when the MF per milliliter is relatively low, and in most cases, these two methods also allow for species-level confirmation of the identity. However, PCR tends to be a very costly test, and both PCR and MKT require a greater degree of expertise and time investment to perform than other tests. Even the lowest performance tests, including the thin smear and hematocrit tube methods, can reliably detect MF at very high burdens; although, caution should be advised when using low reliability methods, since there is a greater likelihood of failing to identify MF-positive dogs. Conclusions Microfilaria (MF) testing is an essential part of heartworm diagnosis and screening in dogs, and test selection should balance practical factors such as cost and time investment with the patient’s risk of infection based on prevention status and history, clinical signs, and antigen testing results. This approach to MF testing will help minimize cost while avoiding failure to detect MF in infected dogs, especially when MF burden is low. Graphical Abstract

Improving access to melarsomine therapy: treating canine heartworm infection in a high-volume, outpatient community clinic setting

BackgroundModels that provide high-quality veterinary care for more affordable prices are emerging, but not well documented outside of wellness and preventative care. Effective treatment guidelines for heartworm disease have been developed by the American Heartworm Society; however, not all owners are able to access treatment due to the high costs associated with sick and emergency care services.MethodsTo increase access to high-quality adulticidal treatment of canine heartworm disease, we developed and implemented a technician-leveraged heartworm treatment protocol for high-volume, outpatient community clinic settings based on the American Heartworm Society guidelines. Modifications were few and included limited pre-treatment blood work, pre-injection sedation, post-injection pain medication, and a reduced exercise restriction period. We monitored retention rates for 556 dogs throughout treatment, evaluated treatment success (defined as no antigen detection 9 months post treatment) for patients that returned for post-treatment antigen testing, and reported on adverse reactions and therapy adherence throughout treatment.ResultsOf the patients that began adulticide therapy, 539/556 (97%) successfully completed the three-injection series. No microfilariae were detected in 99% (428/433) of those who returned for post-injection microfilaria testing. Among those that returned for or reported the results of post-injection antigen testing, no antigen was detected for 99% (245/248) and no microfilariae were detected for 99.5% (200/201). During the course of treatment, 483/539 (90%) of patients experienced at least one adverse reaction, with the most frequently reported types being behavioral and injection site reactions. 25/539 (4.6%) of owners sought additional medical care for adverse reactions at some point during the treatment course. The overall mortality rate was 1.3% (7/556).ConclusionsThis study represents the first evaluation of a heartworm treatment protocol optimized for implementation in a high-volume, outpatient community clinic setting. Our findings align with those previously reported in private practice or tertiary referral centers, illustrating that through the inclusion of pre-treatment blood work, employing short-acting or reversible sedatives, ensuring proper analgesia, minimizing the use of ancillary diagnostics, reducing the duration of in-clinic monitoring while focusing on outpatient care, and maximizing technician involvement, we can deliver effective and safe melarsomine therapy at a more affordable cost to pet owners.Graphical

Efficacy and tolerability of the American Heartworm Society therapeutic protocol in dogs affected by heartworm disease without caval syndrome.

The American Heartworm Society medical protocol represents the current standard of therapy for canine heartworm disease without caval syndrome. However, data on the tolerability of this protocol are limited. This study aimed to describe efficacy and prevalence of possible treatment-related side effects in dogs with heartworm disease treated using the American Heartworm Society protocol. For this retrospective multi-centre cohort study, dogs diagnosed with classes 1 to 3 heartworm disease that completed the American Heartworm Society medical protocol were searched in four medical databases. Demographic, clinical, diagnostic, therapeutic and outcome data, including the number and type of possible treatment-related side effects, were retrieved. Thirty-five dogs were included. The median age and bodyweight were 6 years (1 to 13 years) and 17.3 kg (4.9 to 50 kg), respectively. Heartworm disease was classified as classes 1, 2 and 3 in 20 of 35, 11 of 35 and four of 35 dogs, respectively. In addition to the therapeutic recommendations of the American Heartworm Society, eight of 35 dogs underwent sedation to favour melarsomine administration, and 30 of 35 received ice at the injection site. After adulticide therapy, all dogs were hospitalised with cage rest [median time 12 hours (6 to 48 hours)]. All dogs survived the treatment. All dogs with long-term follow-up (32/35) became negative. Furthermore, treatment-related side effects were rare, mild and rapidly recovered without the need for supporting therapies; these included depression/lethargy (4/35 dogs), cough (2/35 dogs) and lameness, pain and gastrointestinal signs (1/35 dog each). The American Heartworm Society medical protocol is efficient and safe in dogs with classes 1 to 3 heartworm disease.

Effects of doxycycline dose rate and pre-adulticide wait period on heartworm-associated pathology and adult worm mass

BackgroundThe American Heartworm Society canine guidelines recommend treatment with doxycycline prior to adulticide administration to reduce levels of Wolbachia and its associated metabolites, which are known to be a leading cause of pulmonary pathology. Studies have determined that doxycycline administered at 10 mg/kg BID for 28 days is an effective dose for eliminating Wolbachia, but what has not been determined is the clinical relevance of this elimination. The current guidelines also recommend a 30-day wait period following administration of doxycycline to allow for clearance of metabolites, such as Wolbachia surface protein, and for further reduction in heartworm biomass before administration of adulticide. Reducing the doxycycline dose and eliminating the wait period may carry practical benefits for the animal, client, and practitioner.MethodsTo investigate these treatment practices, Dirofilaria immitis adults were surgically transplanted into each of 45 dogs, which were divided into nine study groups of five dogs each. Seventy-five days after transplantation, two groups each were administered 5, 7.5, or 10 mg/kg BID doxycycline orally for 28 days and 6 µg/kg ivermectin monthly, with three untreated groups serving as controls. Study animals were necropsied and examined prior to treatment as well as 30 and 60 days post-treatment.ResultsMean worm weight was unaffected by dosage but exhibited a significant increase at 30 days and significant decrease at 60 days post-treatment, including in control groups. Histopathology lesion scores did not significantly differ among groups, with the exception of the lung composite score for one untreated group. Liver enzymes, the levels of which are a concern in doxycycline treatment, were also examined, with no abnormalities in alanine aminotransferase or alkaline phosphatase observed.ConclusionsNo consistent worsening of tissue lesions was observed with or without the AHS-recommended 30-day wait period, nor did reduced dosages of doxycycline lead to worsening of pathology or any change in efficacy in depleting worm weight. Mean worm weight did significantly increase prior to, and decrease following, the wait period. Future work that also includes adulticide treatment (i.e. melarsomine) will study treatment recommendations that may improve both animal health and owner compliance.Graphical abstract

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

Long-term evaluation of viability of microfilariae and intravenously transplanted adult Dirofilaria immitis in microfilaremic dogs treated with low-dose, short- and long-treatment regimens of doxycycline and ivermectin

BackgroundMicrofilarial (mf) counts were monitored over 21.3 months for any rebound that might occur in counts, and adulticidal efficacy was assessed following administration of low dosage with short- and long-treatment regimens of doxycycline and ivermectin to heartworm-microfilaremic dogs.MethodsTwelve heartworm-naïve beagles infected with 10 pairs of adult Dirofilaria immitis by intravenous transplantation were randomly allocated to three groups of four dogs. All treatments started on day 0. On day 0, Group 1 (short-treatment regimen) received doxycycline orally at 10 mg/kg once daily for 30 days plus ivermectin orally (minimum, 6 mcg/kg) on days 0 and 30. Group 2 (long-treatment regimen) received doxycycline orally at 10 mg/kg once daily until individual dogs became mf-negative (72–98 days) and ivermectin every other week until individual dogs became mf-negative (6–7 doses). Group 3 was the untreated control. Mf counts and antigen (Ag) tests were conducted. Dogs were necropsied for recovery and enumeration of heartworms on day 647.ResultsDay −1 mean mf counts were 15,613, 23,950, and 15,513 mf/ml for groups 1, 2, and 3, respectively. Mean counts for Groups 1 and 2 declined until days 239 and 97, respectively, when all were negative. Group 3 had high mf counts throughout the study. There was not a rebound in mf counts in any of the treated dogs after they became amicrofilaremic. All dogs in group 1 and group 3 were Ag-positive throughout the study and had at least one live female worm at necropsy. All dogs in treated Group 2 were positive for Ag through day 154, but were antigen-negative on days 644 and 647, as all had only male worms. Mean live adult worm recoveries for Groups 1, 2, and 3 were 6.8 (range, 5–8), 3.3 (range, 1–6), and 16.0 (range, 14–17), respectively, with a percent reduction in adult worm counts of 57.5% for Group 1 and 79.3% for Group 2.ConclusionsThese data lend support to the use of the American Heartworm Society Canine Guidelines for adulticide therapy recommending the initiation of doxycycline plus a macrocyclic lactone (ML) at the time of the heartworm-positive diagnosis.Graphical

Further variation of the adulticide protocol for the treatment of canine heartworm infection: can it be even shorter and cost less?

BackgroundThis retrospective study evaluated modified three-dose melarsomine treatment protocols in a shelter setting and compared them to the American Heartworm Society (AHS)-recommended protocol.MethodsAs compared with the AHS protocol, the shelter protocols utilized doxycycline 10 mg/kg once daily (SID) or twice daily (BID), and varied the time from initiation of doxycycline (day 1) to the first melarsomine injection (M1). Dogs were retrospectively grouped based on the shelter’s current protocol (M1 on day 14; Group A) and the AHS protocol (M1 on day 60; Group C), allowing a week on either side of the target M1 day. Treatments that fell outside these ranges formed two additional treatment groups (Groups B and D). Respiratory complications were defined as respiratory signs requiring additional treatment, and were statistically compared for Groups A and C. New respiratory signs and gastrointestinal (GI) signs were compared between dogs receiving SID or BID doxycycline.ResultsOne hundred fifty-seven dogs with asymptomatic or mild heartworm disease at presentation were included. All dogs survived to discharge. There was no statistically significant difference between Groups A (n = 79) and C (n = 27) for new respiratory signs post-melarsomine (P = 0.73). The time to M1 for 14 dogs that developed new respiratory signs was a median of 19 days, compared with 22 days for 143 dogs without new respiratory signs (P = 0.2). Respiratory complications post-melarsomine were uncommon. New respiratory signs post-melarsomine occurred in 10/109 (9.2%) dogs receiving SID doxycycline and 4/48 (8.3%) dogs receiving BID doxycycline (P > 0.999). GI signs prior to M1 were recorded for 40/109 (36.7%) dogs receiving SID doxycycline and 25/48 (52.1%) receiving BID doxycycline (P = 0.08). Forty-four follow-up antigen test results were available; all tests performed > 3 months after the third melarsomine injection were negative.ConclusionsThis study provided support for initiating melarsomine after 14 days of doxycycline and for a lower doxycycline dose. Shorter and less expensive treatment protocols can increase lifesaving capacity and improve quality of life for shelter dogs by reducing the duration of exercise restriction and length of stay.Graphical

Application of Moxidectin and Ultrasound-Aided Injection of Melarsomine During the American Heartworm Society Recommended Treatment Protocol in Dirofilaria immitis Infected Dogs.

The American Heartworm Society (AHS) recommends the three-dose alternate melarsomine therapeutic regimen, together with a macrocyclic lactone (ML) to reduce new infections and eliminate susceptible larvae and doxycycline against Wolbachia bacteria. Till now, only reports on ivermectin as an ML exist in the frame of this protocol. Between 2014 and 2020, the AHS protocol was used in 44 heartworm-positive dogs. Microfilaremic dogs were pretreated with prednisolone and clopidogrel for 1 week before the first moxidectin application. Moxidectin was applied on the 1st, 30th, 60th, and 90th therapeutic days. On the first day, dexamethasone and chloropyramine were used to avoid potential adverse effects caused by the destroyed microfilariae. During the 1st-28th days, doxycycline 10 mg/kg BID was given with probiotics. Adult heartworms were destroyed with melarsomine on the 60th, 90th, and 91st days. Butorphanol and dexamethasone were given just before melarsomine injections. The depth of the intramuscular injection site was determined by ultrasound examination of the lumbar muscles. From the 60th day, dalteparin was applied for 10 days to decrease the chance of pulmonary thromboembolism. Moxidectin did not cause adverse reactions, even in microfilaremic dogs. Gastrointestinal side effects of doxycycline were observed in three (6%) dogs, they recovered after symptomatic therapy and by lowering the initial dose to 5 mg/kg BID. Transient anorexia and diarrhea were found in one (2%), and coughing and mild dyspnea in one (2%) animal as systemic post-therapeutic complications of melarsomine. No local side effects were observed in 13 (30%) dogs, mild local side effects occurred in 29 (66%) patients, and severe local swelling in 2 (4%) cases. All dogs recovered clinically by the 120th day and no microfilaremia was seen that time. An antigen test performed in 37/44 animals on the 271st day was also negative in all cases.

Pharmacoeconomic Analysis of Heartworm Preventive Compliance and Revenue in Veterinary Practices in the United States.

Background: Heartworm disease (HWD) is a potentially fatal condition caused by the nematode Dirofilaria immitis. It is endemic in North America, and the American Heartworm Society recommends that owned dogs be on a Food and Drug Administration-approved HWD preventive year-round. The objective of this study was to compare the 12-month HWD preventive purchase compliance rates of injectable moxidectin (ProHeart® 6) and the dose equivalent in monthly HWD preventives and their associated economic value to the veterinary hospital.Methods: This study used retrospective anonymized transactional data of 7,926,392 unique dogs from 3,737 companion animal practices across the US for the period 2014–2017. Compliance was defined using American Heartworm Society guidelines. Comparisons were purchases of a 6-month moxidectin injection or six doses of any monthly HWD or HWD combination preventive product, tracked for the next preventive purchase 5–7 months later. Total revenue, HWD prevention cost, 12-month repurchase compliance, and patient retention were calculated. Data were expressed on an annualized basis. Compliance comparisons were calculated based on proportion analysis with the SAS ProbNorm function (SAS 9.4, Cary, NC), using a two-sided t-test, at the 5% level of significance (P < 0.05).Results: At 51.7%, annual compliance with injectable moxidectin was higher than the dose equivalent in monthly HWD preventives, which was 24.4% (P = 0.0001). Eighty-five percent of patients on injectable moxidectin recorded additional transactions during the first visit (average invoice of $161), compared with only 55% of pet owners who purchased monthly HWD prevention (average invoice $141) or monthly HWD combination (average invoice of $171). The average costs of 6 months of HWD preventives were as follows: injectable moxidectin, $48 (29.7% of the total visit invoice); monthly HWD prevention, $45 (31.0% of the total invoice); and monthly HWD combination, 95 (55.6% of the total visit invoice). Finally, dogs receiving injectable moxidectin had a higher proportion of patients with repeat injections within 12 months between 2014 and 2017, with 68% retention rate after 4 years. In comparison, the six-dose monthly HWD cohort retention rate dropped to 55% by 2017.Conclusions: Dogs receiving injectable moxidectin had higher HWD preventive compliance, generated more practice revenue, and had a higher rate of practice retention compared with monthly HWD products.

Optimizing heartworm diagnosis in dogs using multiple test combinations

BackgroundVarious heartworm (HW) diagnostic testing modalities detect products of, or reactions to, different life cycle stages of Dirofilaria immitis. Microfilariae (Mf) can be directly visualized in blood, antigen (Ag) from immature and adult heartworms may be detected on commercial assays, and antibody (Ab) tests detect the host immune response to larval stages. Ag and Mf tests are commonly used in dogs, which frequently carry adult HW infections, but Ab tests have only been validated for use in cats. In some HW-infected dogs, Ag is blocked by immune complexing leading to false-negative results. Heat-treatment (HT) to disrupt these complexes can increase the sensitivity of HW Ag tests. The aim of this study was to compare different methods for diagnosing HW infection in dogs at high risk using individual and paired diagnostic tests, including an exploration of using Ab tests designed for cats to test canine samples.MethodsOne hundred stray adult (≥ 2-year-old) dogs in Florida shelters were tested using Mf, HW Ag, and HW Ab tests (feline HW Ab tests currently not commercially validated/approved for use in dogs); two versions of each test platform were used.ResultsFourteen dogs tested positive using point-of-care (POC) Ag tests; an additional 2 dogs tested positive with microtiter well assay, and an additional 12 dogs tested positive using HT Ag testing. For individual tests, Ag test sensitivity/specificity compared to HT Ag was 50–57%/100%, and Ab tests were 46–64%/82–94%. Sensitivity estimates for individual tests were higher when comparing to non-HT Ag. Pairing POC Ag tests with Mf tests improved sensitivity without loss of specificity, while pairing POC Ag and Ab tests modestly increased sensitivity at the expense of specificity.ConclusionsScreening dogs for HW infection using both POC Ag and Mf detection, which is recommended by the American Heartworm Society, improved diagnostic performance in this study compared to single Ag test use, but may have missed more than one in four infected dogs. The need to improve access to highly accurate, rapid, and inexpensive large-scale HW testing for dogs in animal shelters remains largely unmet by current testing availability. The development of practical and validated protocols that incorporate heat or chemical treatment to disrupt Ag-Ab complexes in POC testing or decreasing the cost and time required for such testing in reference laboratories might provide solutions to this unmet need. Similar studies performed in countries where the prevalence of parasites such as D. repens or A. vasorum is different to the USA could potentially yield very different positive predictive values for both HT and non-HT Ag tests.Graphical abstract

Heartworm control in Grenada, West Indies: Results of a field study using imidacloprid 10% + moxidectin 2.5% and doxycycline for naturally-acquired Dirofilaria immitis infections

Canine heartworm disease (CHD) results from infection with Dirofilaria immitis and while it is of global concern, it is most prevalent in tropical climates where conditions support the parasite and vector life cycles. Melarsomine dihydrochloride is the sole treatment for CHD recommended by the American Heartworm Society. However, in cases where cost or access to melarsomine precludes treatment of an infected dog, therapeutic alternatives are warranted. This randomized, controlled field study evaluated the adulticidal efficacy of a combination therapeutic protocol using 10 % imidacloprid + 2.5 % moxidectin spot-on and a single 28-day course of doxycycline and compared with that of a 2-dose melarsomine dihydrochloride protocol. Of 37 naturally-infected domestic dogs with class 1, 2 or early class 3 CHD enrolled in the study, 30 were evaluated for a minimum of 12 months. Seven dogs were withdrawn due to canine ehrlichiosis, non-compliance, or wrongful inclusion. Dogs were randomly assigned to a control (CP, n = 15) or investigational (IVP, n = 15) treatment group. CP dogs received two injections of melarsomine dihydrochloride (2.5 mg/kg) 24 -hs apart and maintained on monthly ivermectin/pyrantel. IVP dogs were treated with oral doxycycline (10 mg/kg twice daily for 28 days) and topical 10 % imidacloprid + 2.5 % moxidectin once monthly for 9 months. Dogs were evaluated up to 18 months - monthly for the first 9 months, then every 3 months. Parasiticidal efficacy was based on antigen status using the IDEXX PetChek® 34 Heartworm-PF Antigen test. By month 18, antigen was not detected in any study dog except one from the IVP group. One other IVP dog was persistently antigenemic and treated with melarsomine at month 12 according to the initial study protocol. Mean antigen concentration (based on optical density) decreased more rapidly in the CP group and by month 15 was 0.11 for the IVP and 0.07 for CP groups, with equivalent median concentrations (0.04) in both groups. Conversion following heat-treatment of antigen-negative samples occurred frequently and at similar rates in both treatment groups. Based on the bias of diagnostic tests towards detection of female worms, we conclude that monthly application of 10 % imidacloprid + 2.5 % moxidectin for 9 months combined with a course of doxycycline twice daily for 28 days resulted in effective therapy against female adults in CHD. This therapeutic option may be particularly useful in cases where financial constraint or access to melarsomine precludes treatment of an infected individual. This study was supported by Bayer Animal Health.

The American Heartworm Society and Association of Shelter Veterinarians’ 2019 Shelter Heartworm Management Practices Survey

The primary objective of the 2019 Shelter Heartworm Management Practices Survey was to identify the current practices employed for the prevention, diagnosis, and management of heartworm disease in animal shelters. Survey questions were developed to collect the information of interest for dogs, cats, and ferrets under the care of sheltering organizations along with demographic information about these organizations. An electronic survey was distributed to the American Heartworm Society and Association of Shelter Veterinarians’ mailing lists in February-March 2019. Cross-posting of the survey on relevant lists was encouraged. The survey invitation was directed toward veterinarians delivering care in an animal shelter and excluded practitioners providing services for shelters, shelter animals, or recently adopted animals within a private practice setting. A total of 242 survey responses were received representing private humane societies/societies for the prevention of cruelty to animals (39.7%), municipal animal control facilities (24.9%), traditional (public-private partnership) shelters (21.7%), foster-based organizations (7.9%), sanctuaries (2.6%), and breed/species-specific rescues (2.1%). The largest proportion of respondent shelters were from the southeastern United States (36.9%) and over two-thirds of the shelters had full-time shelter veterinarians (69.0%). Most respondents provided monthly heartworm preventives for dogs (81.1%) and used oral ivermectin products per product labels (67.2%) to do so. Extralabel use of preventives was reported by 21.3% of respondents. Most respondents tested all dogs >6 months of age for heartworm infection (81.9%) and did so during the intake examination (56.6%) with antigen tests (92.2%). At least some heartworm positive dogs were treated by most respondents (93.1%) using a 3-dose (42.5%) or 2-dose (37.9%) melarsomine protocol. Non-arsenical treatment protocols were utilized by 16.8% of respondents. Protocols typically included adjunctive treatment with doxycycline or minocycline (92.3%), prednisone (71.3%), and macrocyclic lactones (58.6%). Nearly two-thirds of respondents housed dogs within foster homes after treatment (65.1%). Heartworm prevention was provided by 29.9% of respondents that admitted cats and 9.9% of those that admitted ferrets. These results can be used to identify challenges to meeting recommended practices regarding prevention, diagnosis, and treatment of heartworm disease in the animal shelter setting and to target operational and educational resources toward the areas of greatest need.

Non-Arsenical heartworm adulticidal therapy using topical moxidectin-imidacloprid and doxycycline: A prospective case series

This prospective case series evaluated the adulticidal efficacy of topical 10 % moxidectin/2.5 % imidacloprid (M/I; Advantage Multi®, Bayer, Shawnee Mission, KS, USA) and doxycycline in dogs with naturally occurring heartworm infection (HWI). Twenty-two dogs with HWI whose owners declined melarsomine were treated with M/I at the preventive dosage twice monthly for 90 days then monthly thereafter and doxycycline (median [interquartile range; IQR] dosage 12.6 [12.0–16.1] mg/kg/day) for the first 15 days. Although strict activity restriction was not imposed, owners were asked to prevent their dogs from exercising strenuously. This protocol was referred to as the MOXY protocol. Antigen testing was performed every 30–60 days, until dogs had ‘no antigen detected’ (NAD). Twenty-one of the 22 dogs ultimately converted to NAD by 434 days (median [IQR]), 234 (179–303). One dog remained positive 701 days after MOXY initiation and was considered a treatment failure. All sera which converted to NAD on HW antigen testing were retested after heat-treatment. Twelve dogs had NAD on the heat-treated test on the same day as having their first NAD on the conventional test. Six of 9 dogs testing positive after heat-treatment were retested and all 6 had NAD on a heat-treated test within 2–3 months. Microfilaremia was cleared in all 8 dogs re-tested. Four dogs required treatment for cough, thought due to heartworm (HW) death, an average of 89 days after initiation of MOXY. This cough was most likely due to pneumonitis with heartworm-pulmonary thromboembolism. One dog required hospitalization for 24 -h and recovered fully with corticosteroid therapy and supportive care and 2 dogs were treated in an outpatient fashion with steroids. The MOXY protocol was tolerated and 96 % (21/22) of dogs converted to NAD, though 2 dogs required greater than 1 year to achieve this result. Nonaresenical-adulticide therapy may result in pneumonitis and heartworm-pulmonary thromboembolism at unpredictable times, potentially months after initiation of macrocyclic lactone therapy and exercise restriction should be considered when using a nonarsenical protocol. Although not currently recommended by the American Heartworm Society (AHS), non-arsenical strategies are in use and the goal of this study was to evaluate the efficacy, duration of therapy, and safety of an accelerated dosing protocol of M/I with doxycycline.

Assessing the potential cross-reactivity using a commercial heartworm ELISA kits of serum from dogs naturally infected with Onchocerca lupi

Onchocerca lupi is an emerging zoonotic parasite of dogs, endemic to the southwestern USA and areas of the Old World. Currently, there are no specific serological diagnostic tests able to detect O. lupi infection. Recent literature has demonstrated that commercially available heartworm antigen tests, despite being highly sensitive, may cross-react with infections by other filarid nematodes. There is no information on potential cross-reactivity of such tests in serum of dogs infected with O. lupi. Our objective was to assess serum samples of dogs naturally-infected with O. lupi for potential cross-reactivity before and after heat-treatment using a commercial heartworm ELISA kit. We obtained serum from 23 dogs naturally-infected with O. lupi. These dogs presented with ocular disease, and were consulted to schedule either surgical removal of ocular nodules due to infection or enucleation. Samples were tested in triplicate using the DiroCHEK® Heartworm Antigen Test kit (Synbiotics Corporation, Zoetis, Kalamazoo, MI, USA) following the manufacturers’ protocol pre- and post-heat-treatment. Samples were heat-treated using a dry heat block at 103 °C for 10 min and then centrifuged at 1818×g for 20 min. Out of a total of 23 dogs, 19 (82.6 %) had no antigen detected regardless of heat-treatment, three dogs tested positive before and after heat-treatment, and a single dog turned positive after heat-treatment. These three dogs that were positive before and after heat-treatment were confirmedly co-infected with Dirofilaria immitis by the veterinarians responsible for these cases, and we were unable to get the history or follow up with the dog that turned positive post-heat-treatment only. Our data suggest that O. lupi infections should not result in false-positives when using the DiroCHEK® in dog serum, before or after heat-treatment. Dogs with clinical ocular onchocercosis that test antigen-positive in DiroCHEK® are likely co-infected with D. immitis, and should be further tested, including evaluation of microfilariae in blood and diagnostic imaging. If heartworm infection is confirmed, the animals should be enrolled in the recommended treatment protocol in accordance to the guidelines of the American Heartworm Society or other local organizations.

Efficacy of imidacloprid 10%/moxidectin 2.5% spot on (Advocate®, Advantage Multi®) and doxycycline for the treatment of natural Dirofilaria immitis infections in dogs

Heartworm infection (also known as dirofilariosis due to Dirofilaria immitis) in dogs causes chronic pulmonary disease that, if left untreated, can lead to right-side congestive heart failure. Currently, the only registered drug for adulticide therapy in dogs with heartworm disease (HWD) is melarsomine dihydrochloride. The recent targeting of the bacterial endosymbiont Wolbachia, through antibiotic therapy of the infected host, has offered an interesting alternative for the treatment of HWD. Recent reports of the adulticide activity of an ivermectin/doxycycline combination protocol has lead the American Heartworm Society (AHS) to include in its guidelines that, in cases where arsenical therapy is not possible or is contraindicated, a monthly heartworm preventive along with doxycycline for a 4-week period might be considered. In the present study, 20 dogs with confirmed natural D. immitis infection were included following owner consent. Fourteen dogs were treated with a topical formulation containing 10% w/v imidacloprid and 2.5% w/v moxidectin (Advocate®, Advantage Multi®, Bayer), monthly for nine months, associated to doxycycline (10 mg/kg/BID) for the first 30 days. Six dogs were treated with melarsomine (Immiticide®, Merial) (2.5 mg/kg) at enrollment, followed one month later by two injections 24 h apart.The presence of circulating antigens and the number of microfilariae (mf) were evaluated at the moment of enrollment and then at 1, 2, 3, 4, 5, 6, 7, 8, 12, 18, 24 months post enrollment. Echocardiogram and radiographs were performed at month 0, 6, 12, 18, 24.Monthly moxidectin combined with 30 days of doxycycline eliminated circulating microfilariae within one month, thus breaking the transmission cycle very quickly. Furthermore, dogs treated with the combination protocol started to become negative for circulating antigens at 4 months from the beginning of treatment and all except one were antigen negative at 9 months. All dogs treated with melarsomine were antigen negative by 5 months from the beginning of the treatment. No dogs showed worsening of pulmonary patterns or criteria indicative of pulmonary hypertension 12 to 24 months after. For the criteria mf concentration, antigen concentration, radiography and echocardiography at 12, 18 and 24 months the non-inferiority for the moxidectin group could be proven for a non-inferiority margin of 15% for the rate difference. Dogs treated with moxidectin and doxycycline became negative for microfilariae and antigens sooner when compared to melarsomine in the present study and to dogs treated with doxycycline combined with ivermectin in studies previously published.

Efficacy and side effects of doxycycline versus minocycline in the three-dose melarsomine canine adulticidal heartworm treatment protocol

BackgroundThe American Heartworm Society currently recommends the use of a monthly macrocyclic lactone, a 28-day course of 10 mg/kg doxycycline BID, and the 3-dose protocol of melarsomine dihydrochloride for the treatment of canine heartworm disease. Doxycycline is necessary for the reduction of the bacterium Wolbachia, found in all heartworm life-stages. Previous price increases and decreasing availability prompted us to evaluate alternative tetracycline antibiotics, i.e. minocycline, for the reduction of Wolbachia during canine heartworm treatment.MethodsThirty-two heartworm-positive dogs were randomized to receive 10 mg/kg or 5 mg/kg of either doxycycline or minocycline for 28 days BID, for a total of 8 dogs per experimental group. All dogs received 6 months of Heartgard Plus® (ivermectin/pyrantel) and the 3-dose protocol of 2.5 mg/kg melarsomine dihydrochloride. Blood samples were collected prior to the initiation of treatment, every 7 days throughout tetracycline treatment, and then monthly thereafter until the dog tested negative for the presence of heartworm antigen. DNA was isolated from circulating microfilarial samples and qPCR was performed on each sample.ResultsA greater number of dogs in the 10 mg/kg doxycycline and minocycline treated groups experienced gastrointestinal side effects as compared to the 5 mg/kg doxycycline and minocycline treated groups. All eight dogs in the 10 mg/kg doxycycline-treated group tested negative for the presence of Wolbachia DNA by 28 days post-tetracycline treatment. A total of two dogs in both the 5 mg/kg doxycycline- and 10 mg/kg minocycline-treated groups and three dogs in the 5 mg/kg minocycline-treated group remained positive for the presence of Wolbachia DNA by the end of tetracycline treatment.ConclusionsNo lung pathology was assessed in this clinical trial, therefore the clinical effect of the remaining Wolbachia DNA in the 10 mg/kg minocycline-, 5 mg/kg doxycycline- and 5 mg/kg minocycline-treated groups cannot be determined. Owner compliance in the proper administration of these tetracyclines may be impacted by the increased severe gastrointestinal side effects reported for the 10 mg/kg doxycycline- and minocycline-treated groups. We recommend that veterinarians prescribe the recommended 10 mg/kg doxycycline for canine heartworm treatment and reduce the dosage to 5 mg/kg in cases of severe gastrointestinal side effects in order to improve owner compliance in administration of medications.

Approaches to Canine Heartworm Disease Treatment Among Alumni of a Single College of Veterinary Medicine.

This descriptive study was designed to ascertain the current heartworm treatment strategies employed by veterinary graduates of a single college of veterinary medicine, to assess the frequency with which each of these treatment strategies is prescribed, and to report the motivation behind the use of these treatment strategies. A survey containing a combination of multiple-choice and open-ended questions was distributed via e-mail with an online link during 2013 to graduates of the University of Georgia College of Veterinary Medicine. Demographic data and opinions regarding treatment for cases of canine heartworm disease (HWD) were obtained, and motivation for recommending different treatment strategies was assessed. Nearly all 170 respondents (99%) indicated that they recommend melarsomine dihydrochloride for first-line treatment of canine HWD. Exercise restriction (80%) and monthly heartworm preventive (75%) were components of the treatment approach to HWD with no clinical signs. The majority of respondents (74%) indicated that when first-line treatment recommendations were declined, they endorsed long-term administration of ivermectin (i.e., "slow-kill" method) despite current American Heartworm Society guidelines that recommend against the use of long-term macrocyclic lactone administration for the monotherapy treatment of canine HWD. Respondents also indicated that owners' financial concerns frequently result in modification of HWD treatment. Routine inclusion of exercise restriction is commonly, but not universally, utilized and may represent an opportunity for improvement in the management of this disease. In addition, when first-line recommendations for heartworm disease treatment are declined, a two-dose melarsomine protocol instead of the slow-kill method should be considered.

Increasing incidence of Dirofilaria immitis in dogs in USA with focus on the southeast region 2013\u20132016

BackgroundA recent American Heartworm Society (AHS) survey on the incidence of adult heartworm infections in dogs in the United States of America showed a 21.7% increase in the average cases per veterinary clinic from 2013 to 2016. The analysis reported here was performed to see if heartworm testing results available via the Companion Animal Parasite Council (CAPC) aligned with the AHS survey and whether changes in heartworm preventive dispensing accounts for the increased incidence. The resistance of Dirofilaria immitis to macrocyclic lactones (MLs) has been previously reported.MethodsAn analysis of 7–9 million heartworm antigen tests reported annually to the Companion Animal Parasite Council (CAPC) from 2013 to 2016 was conducted and compared to the 2016 AHS survey. A state-by-state analysis across the southeastern USA was also performed. National heartworm preventive dispensing data were obtained from Vetstreet LLC and analyzed. All oral, topical and injectable heartworm preventives were included in this analysis, with injectable moxidectin counting as six doses.ResultsPositive antigen tests increased by 15.28% from 2013 to 2016, similar to the 21.7% increase reported by the AHS survey. Incidence in the southeastern USA increased by17.9% while the rest of USA incidence increased by 11.4%. State-by-state analysis across the southeastern USA revealed an increased positive test frequency greater than 10% in 9 of 12 states evaluated. During this time, the overall proportion of dogs receiving heartworm prophylaxis remained relatively unchanged. Approximately 2/3 of the dogs in the USA received no heartworm prevention each year.ConclusionThese CAPC data show the rate of positive heartworm tests increasing significantly (P < 0.0001) in the USA from 2013 to 2016, with a higher rate of increase in the southeastern USA than nationally. Only 1/3 of dogs in the USA were dispensed one or more doses of heartworm prevention annually by veterinarians, averaging 8.6 monthly doses/year. Veterinarians and pet owners should work together to follow CAPC and AHS guidelines to protect dogs from infection with D. immitis. Lack of preventive use and the emergence of heartworm resistance to MLs could both be impacting the increased rate of positive heartworm tests in dogs.

Evaluation of cardiopulmonary and inflammatory markers in dogs with heartworm infection during treatment with the 2014 American Heartworm Society recommended treatment protocol

BackgroundHeartworm disease in dogs is a life-threatening parasitic disease. Although adulticide treatment with melarsomine has been proven to be the most effective, complications associated with adulticide treatment are major concerns for clinicians.MethodsThis study evaluated the change in levels of D-dimer, interleukin-6, C-reactive protein and cardiac troponin I in 12 dogs with different severities of heartworm infection treated by the American Heartworm Society (AHS) recommended protocol during the treatment period. The serum levels of several markers were measured on the day of diagnosis (T-60), before the initiation of melarsomine therapy (T0), 1 day after the first injection (T1), 1 week after the first injection (T7), 1 month after the first injection (T30), 1 day after the second injection (T31), 1 day after the third injection (T32), 1 week after the third injection (T39), 1 month after the third injection (T62), 2 months after the third injection (T92), 3 months after the third injection (T122), and 6 months after the third injection (T182).ResultsThe serum levels of these markers were significantly different at the test time point after melarsomine treatment and also differed significantly according to the stage of heartworm disease in the dogs.ConclusionThis study found that monitoring of inflammatory and hemostatic markers in dogs with heartworm disease being treated with melarsomine might be beneficial in predicting the clinical outcomes and complications associated with melarsomine treatment.

Investigating management choices for canine heartworm disease in northern Mississippi

BackgroundThere are concerns that the chronic use of macrocyclic lactone preventives to kill adult heartworms (“soft-” or “slow-kill”) may have contributed to the development of macrocyclic lactone resistance. This prospective analysis was designed to expand our understanding of currently employed treatment decisions, protocols utilized in a “slow-kill” methodology, and trends in heartworm prevention in a region with concerns about macrocyclic lactone resistance. We tested the hypothesis that practitioners underestimate the actual percentage of heartworm-positive dogs treated with “slow-kill” therapy. Owners’ financial concerns and veterinarians’ efforts at meeting client preferences are the primary reasons for employment of “soft-kill” treatment.MethodsA prospective analysis of dogs determined to be heartworm-positive when presented to a mixed-animal practice in northern Mississippi was conducted for the second quarter of 2016. Client records were scrutinized for heartworm preventive purchase history. Veterinarians in the four-doctor practice completed a questionnaire regarding their beliefs and practices of heartworm treatment.ResultsForty of 321 canine patients tested heartworm-positive with a commercial antigen test kit. Of these, two were considered to be due to possible product failure. The majority (75.0%) of patients received a “slow-kill” method, a percentage greater than that estimated by the practitioners. Patients were equally likely to have received adulticidal treatment as they were to receive no treatment (12.5%). Injectable moxidectin was the most common preventive used in “slow-kill” treatment (80.65%). Client financial concerns were cited as the primary reason for choosing “slow-kill” treatment (79.0%).ConclusionsDespite American Heartworm Society recommendations, clients and veterinarians prefer the “slow-kill” method of heartworm treatment. Trends in patient heartworm preventive history show that poor client compliance remains the predominant explanation for heartworm infection. Thus, consistent use of existing, effective heartworm preventives should be the primary goal in reducing prevalence of heartworm infection, regardless of the recognized threat of resistance. It is also noteworthy that practitioner estimates may be suspect in their accuracy. Further study is needed on the risks and efficacy of “slow-kill” treatment and the effects of different ML preventives for this off-label use.