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Disseminated intravascular coagulation following melarsomine therapy for Dirofilaria immitis (D. immitis) is reported in a 9-year-old female intact pit bull-type dog. The dog had been diagnosed with D. immitis (antigen and microfilaria positive) and treated with imidacloprid, moxidectin, doxycycline and 3 doses of melarsomine over a 92-day period. Seven days after the third melarsomine injection, the patient was presented to her family veterinarian due to right pelvic limb swelling. Prothrombin and activated partial thromboplastin times were prolonged beyond the detectable range. Treatment included vitamin K1 and fresh frozen plasma (FFP) prior to referral to the authors' institution. At this time the patient remained coagulopathic. Further investigations included thoracic radiographs, abdominal ultrasound and an echocardiogram. The patient was administered multiple units of packed red blood cells and FFP, sildenafil, dexamethasone SP, aminocaproic acid and vitamin K1. Repeat CBC approximately 20 h after admission showed persistent anemia and thrombocytopenia. Despite ongoing administration of FFP, a repeat coagulation panel showed worsening of the coagulopathy with prothrombin time of 84.2s [reference interval (RI) 7.0–9.3s], activated partial thromboplastin time >140s (RI 10.4–12.9s) and fibrinogen D. immitis. Potential mechanisms for the coagulopathy are discussed and the case report highlights a rare, but serious complication of adulticide therapy.

Canine heartworm infection, caused by the filarial parasite Dirofilaria immitis, represents a serious and expanding animal welfare concern that is expected to increase due to the effects of climate change and the COVID-19 pandemic. A body of evidence has emerged to support the use of a non-arsenical adulticide treatment protocol, using moxidectin and doxycycline to kill adult heartworms over a prolonged period. While a three-dose protocol using the arsenical drug melarsomine is currently the safest and most effective treatment for heartworm infection, this drug is not available in some countries and is inaccessible for many owners and animal shelters. Moxidectin-doxycycline (moxi-doxy) provides a viable alternative to no treatment at all, in cases where arsenical treatment is not possible. Based on current evidence, the most effective non-arsenical treatment regimen is doxycycline 10 mg/kg PO q 12 or 24 h for 28 days, combined with topical moxidectin at label dose. Moxidectin is repeated monthly until no antigen detected (NAD) status is confirmed. Sustained release injectable moxidectin, in combination with doxycycline, may provide an alternative in remote regions or in settings where significant compliance or accessibility concerns exist, but more studies are needed. In moxi-doxy protocols, doxycycline should be repeated annually until NAD. This review summarizes the safety and efficacy of moxi-doxy, addresses controversies surrounding this treatment approach, and provides detailed recommendations for treatment regimens and post-treatment testing.

The canine heartworm, Dirofilaria immitis, is among the most important parasites of dogs in the United States and worldwide, and may cause severe and potentially fatal disease. Current diagnostic recommendations rely on serological detection of an adult female antigen, and visualization of microfilariae in the blood. Therefore, a reliable diagnosis can be only performed approximately six months post-infection. There is a growing need to characterize novel diagnostic markers that are capable of detecting the early stages of heartworm infection, in special markers associated with third-stage larvae (L3) and fourth-stage larvae (L4). The early detection of infection would guide medical interventions that could impede the development of patent infections and further parasite transmission. We cultured D. immitis L3 and L4 of two laboratorial strains with different susceptibility statuses to macrocyclic lactone drugs in vitro. Excretory/secretory microRNAs were sequenced and analyzed. We identified two miRNA novel candidates secreted abundantly by both L3 and L4 of both strains. These candidates were previously detected in the secretions of other D. immitis stages and one of them was found in the blood of D. immitis-infected dogs. These miRNAs have not been found in the secretions of other nematodes and could be D. immitis-specific diagnostic biomarkers, which could allow for the early detection of infection.

Summary Over the past decade, increasing numbers of autochthonous cases of heartworm infection have been reported in the countries of Eastern Europe where previously only imported cases were described. In this report we have described the first clinical case of Dirofilaria immitis infection in an imported dog in Lithuania. In 2018, a 5-year-old male Spanish greyhound (Spanish galgo) was imported to Lithuania from southern Spain and referred to a small animal veterinary clinic in Vilnius for wellness screening. Circulating microfilariae and female antigens of D. immitis were detected using the Knott’s test and SNAP 4Dx Plus Test (IDEXX Laboratories, Portland, USA). The diagnosis was confirmed using molecular analysis. Treatment according to the guidelines recommended by the American Heartworm Society was applied. This is the first confirmed report of canine heartworm infection in an imported dog in Lithuania. Heartworm-infected dogs transported to North-Eastern Europe from endemic areas could act as microfilarial reservoirs for the local mosquito population, which could increase the risk of spreading the disease.

BACKGROUND: Vector‐borne diseases have an adverse impact on health of dogs, and infected dogs can be sentinels for human infection. Infection with Trypanosoma cruzi, an agent of Chagas disease, causes fatal heart disease in dogs across the southern United States but has been neglected from wide‐scale prevalence studies. OBJECTIVES: To determine the prevalence of exposure to T. cruzi, Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi, and infection with Dirofilaria immitis among dogs in shelters across Texas and to identify risk factors for T. cruzi seropositivity. ANIMALS: Six hundred and eight dogs. METHODS: This repeated cross‐sectional study was performed by collecting blood from ~30 dogs during each of the 3 visits to 7 shelters. We tested serum for antibodies to T. cruzi using 2 tests in series and for antibodies to Ehrlichia spp., Anaplasma spp., and B. burgdorferi and D. immitis antigen using the IDEXX SNAP 4DX Plus point‐of‐care test. DNA was extracted from blood clots and tested for T. cruzi DNA and strain type via quantitative polymerase chain reactions (qPCR). We used logistic regression to assess risk factors. RESULTS: One hundred ten (18.1%) of 608 dogs were seropositive for T. cruzi. Prevalence of exposure to the other vector‐borne agents was: Ehrlichia spp. 3.6%; Anaplasma spp. 6.9%; B. burgdorferi 0.2%; and D. immitis infection 16.0%. Six of 559 (1.1%) dogs were qPCR‐positive for T. cruzi. CONCLUSIONS AND CLINICAL IMPORTANCE: T. cruzi seroprevalence was comparable to D. immitis prevalence and higher than seroprevalence of the tick‐borne pathogens. T. cruzi is an underrecognized health threat to dogs across Texas and possibly other southern states where triatomine vectors are endemic.

Background The current studies compared ProHeart® 12, Heartgard® Plus and Interceptor® Plus for preventive efficacy against JYD-34, a macrocyclic lactone (ML)-resistant strain of Dirofilaria immitis in dogs. Methods In two studies, each using 24 adult beagles, dogs were allocated to four treatment groups (n = 6): placebo-treated control; ProHeart 12 as per label (0.5 mg/kg moxidectin); Heartgard Plus (HGP) as per label (minimum 6 µg/kg ivermectin); and Interceptor Plus (INP) as per label (minimum 0.5 mg/kg milbemycin oxime). In both studies, ProHeart 12 was administered as a single subcutaneous dose on day 0, and HGP and INP were administered orally on days 0, 30, 60, 90, 120 and 150. In Studies 1 and 2, dogs were inoculated with 50 third-stage heartworm larvae (JYD-34 strain) on days −30 and 165, respectively. In Study 2, treatment for both HGP and INP was continued on days 180, 210, 240, 270, 300 and 330. Adult heartworm recoveries were performed on day 185 in Study 1 and on day 360 in Study 2. Results In Studies 1 and 2, all placebo-treated dogs developed adult heartworm infections (geometric mean, 29.9 and 34.9 worms/dog, respectively). A single dose of ProHeart 12 was 100% effective in preventing the development of adult JYD-34 heartworms when treatment was initiated 30 days after heartworm inoculation, while six consecutive monthly doses of HGP and INP were only 10.5% and 14.6% effective, respectively. The mean worm count for the ProHeart 12-treated group was significantly lower (P

Background: Controlling canine vector-borne diseases (CVBD) is a major concern, since some of these diseases are serious zoonoses. This study was designed to determine seropositivity rates in Spain for agents causing the following five CVBD: leishmaniosis (Leishmania infantum: Li), heartworm (Dirofilaria immitis :D i), ehrlichiosis (Ehrlichia canis :E c), anaplasmosis (Anaplasma phagocytophilum/Anaplasma platys: An) and Lyme disease (Borrelia burgdorferi :B b). Methods: Anti-An, -Bb, and -Ec antibodies and the Di antigen were determined using the 4DX SNAP W Test (IDEXX Laboratories) and anti-L. infantum (Li) antibodies using the Leishmania SNAP W Test (IDEXX Laboratories) in blood and/or serum samples. Results: Among 1100 dogs examined, overall seropositivity rates were: Li (15.7%), Ec (5%), An (3.1%), Di (1.25%) and Bb (0.4%). While seropositivity towards Bb and Di was similar in all geographic regions, rates were significantly higher in the east of Spain (8.3%) for An, significantly higher in the north (20%) for Ec, and significantly higher in the Southeast (46.6%) and South (27.4%), and significantly lower in the north (0%) for Li. No statistical associations were observed between sex and the CVBD analyzed (p ≥0.05) while the following associations with other variables were detected: a higher seropositivity to Ec (40%) and Bb (6.7%) in dogs under one year of age compared with adults (p < 0.05); and a higher seropositivity to An and Li in dogs that lived outdoors versus indoors (p=0.01; p < 0.001, respectively). Seropositivity rates of 2.1%, 0%, 1.7%, 0.5% and 4.2% were recorded respectively for An, Bb, Ec, Di and Li in dogs with no clinical signs (n= 556) versus 3.8%, 0.6%, 7.5%, 1.8% and 25.9% for those with signs (n = 507) suggestive of a CVBD. Conclusion: The data obtained indicate a risk for dogs in Spain of acquiring any of the five CVBD examined. Veterinarians in the different regions should include these diseases in their differential diagnoses and recommend the use of repellents and other prophylactic measures to prevent disease transmission by arthropod vectors. Public health authorities also need to become more involved in the problem, since some of the CVBD examined here also affect humans.

Background: Canine heartworm (HW) is endemic in Australia. Prevention usually involves monthly topical or oral preventives, or annual injections of extended-release moxidectin (ProHeart SR-12*1), hereafter referred to as injectable moxidectin (IM). Poor compliance can leave dogs susceptible to infection. This pharmacoeconomics study used retrospective transactional data from 52 Australian veterinary practices to examine the economic value of compliance, revenue, and patient retention associated with veterinarian-sourced canine HW prevention.Methods: This longitudinal descriptive study utilized anonymized transaction records of 228,185 dogs identified to have visited a veterinary practice at least twice in the period 2010–2015. Purchase compliance against a benchmark of 12 months HW protection per year was measured for IM or monthly HW (MHW) preparations each year and for consecutive years. The average annual cost per dog by preventative modality was also determined.Results: Between 2010 and 2015, of the 228,185 dogs identified, 73.0% recorded either zero or one purchase of HW preventive from their veterinary clinic; 18.7% recorded at least two IM purchases, and 10.6% purchased MHW prevention at least twice. Single-year purchase compliance was 92.8–96.9% for IM vs. 26.9–36.5% for dogs receiving MHW products. Consecutive-year purchase compliance was 76.7% for IM and 24.4% for MHW medications. Dog owners spent $AU108.29/dog/year (Australian dollars) on IM vs. $AU131.96/dog/year on MHW prevention products, which may have treated other parasites concurrently, although repeat MHW purchasers only purchased enough to cover an average of 7.2 months per year. Dogs recording at least two HW prevention transactions generated more revenue for veterinary practices/dog/year compared to dogs with less than two. Finally, dogs receiving IM, especially those that started at Conclusions: In the 5 years from 2010 to 2015, 73% of dog owners who visited a veterinary practice at least twice made less than two purchases of HW preventatives from the veterinary practice. For those with at least two preventative purchases, 76.7% of dogs receiving IM and 24.4% of dogs prescribed with MHW products purchased enough doses to provide continuous protection over the observation period.

Background Dirofilaria immitis is a life-threatening nematode spreading globally. Arsenical treatment is currently recommended for removal of adult worms. However, arsenical treatment is not available in some countries, and there are dogs that cannot tolerate the rapid kill of adult worms; therefore, alternative adulticide slow-kill treatments are needed. Criticisms against the use of these alternative protocols include the potential for allowing disease to progress and for the development of ML-resistant worms. Methods The efficacy of a protocol that includes semi-annual doses (i.e. every 6 months) of commercially available extended-release injectable moxidectin suspension (ProHeart® SR-12) with 30-day oral administration of doxycycline was studied in 20 dogs with naturally occurring D. immitis infections. Each dog received treatment with ProHeart® SR-12 (0.5 mg moxidectin/kg) by subcutaneous injection and oral doxycycline (10 mg/kg/bid × 30 days) every 6 months until two consecutive negative antigen test results were obtained. Pulmonary and cardiac evaluations were performed by radiographic and echocardiographic parameters. Physical examinations, complete blood counts, clinical chemistry profiles, microfilariae and antigen tests were performed periodically. Results At enrollment, all dogs were positive for D. immitis antigen and 18 were microfilaremic. On day 30, microfilaremia counts decreased, and all dogs became amicrofilaremic by day 150. On day 180, 11 dogs were antigen-negative, and 7 more became negative by day 360. The two remaining antigen-positive dogs converted to negative by day 540 or 810. All antigen tests performed 180 days after the first negative test were negative. There was no decline in cardiac performance of the dogs throughout the study. Overall, pulmonary clinical conditions, presence of worms by echocardiography, and enlargement of caudal and main pulmonary arteries improved after treatment. Physical examinations, complete blood count results, and clinical chemistry profiles were within normal reference values. Respiratory conditions were improved, no damage to the heart was observed, and the treatment protocol was well tolerated by the animals. Conclusions This alternative adulticide treatment was efficacious and well tolerated in naturally infected dogs. The injectable formulation provides the advantage of having veterinarians able to administer, monitor, and assess the efficacy and condition of the dog throughout the treatment and post-treatment periods.

Background The adult worms of Angiostrongylus vasorum reside in the pulmonary artery of dogs and can lead to cardiac, respiratory, and central neurologic signs. Due to luminal obstruction and perivascular inflammation of the pulmonary artery branches, pulmonary hypertension can arise. Pulmonary hypertension, in turn, can lead to severe damage of the right-sided cardiac structures, leading to right ventricular remodeling and tricuspid valve regurgitation. Case presentation An 8-year-old neutered female English Cocker Spaniel was presented to the author’s institution because of abdominal distention and exercise intolerance. Ascites caused by congestive right-sided heart failure was found to be responsible for these problems. The underlying etiology of the right-sided heart failure was a severe pulmonary hypertension caused by Angiostrongylus vasorum infection. Echocardiography revealed, in addition to a severe concentric and eccentric right ventricular hypertrophy, right atrial and pulmonary trunk dilation, severe tricuspid valve regurgitation, and a systolic flail of the anterior leaflet of the tricuspid valve, resulting from ruptured chordae tendineae. As a coincidental finding, a congenital mitral stenosis was found. Oral therapy was initiated with daily administration of fenbendazole for 2 weeks along with daily administration of oral sildenafil until the re-check examination. At the 6-week re-check the dog showed full clinical and partial echocardiographic recovery, and both the blood antigen test for Angiostrongylus vasorum and the fecal Baermann larva isolation test were negative. When the sildenafil therapy was ceased after tapering the daily dosage, the owner reported recurrence of abdominal distension. Re-starting the sildenafil therapy resulted in resolution of this problem. The dog was reported to be clinically healthy with daily sildenafil administration 7 months after the initial presentation. Conclusions The present case report describes a dog where angiostrongylosis led to congestive right-sided heart failure resulting from severe pulmonary hypertension. The secondary right ventricular eccentric hypertrophy together with suspected papillary muscular ischemia were the suspected cause of the ruptured major tricuspid chordae tendineae, which led to a severe tricuspid valve regurgitation. Despite eradication of the worms, the severe pulmonary hypertension persisted. Treatment with daily oral sildenafil, a pulmonary arterial vasodilator, was enough to keep the dog free of clinically apparent ascites.

Background Dirofilaria immitis is a filarial parasite of dogs that can cause serious or fatal cardiopulmonary disease. Three studies were conducted to evaluate the efficacy and safety of monthly treatment with moxidectin in a chewable tablet product in combination with sarolaner and pyrantel to prevent heartworm disease in dogs after experimental challenge and in a clinical field study in the USA. Methods In two laboratory studies, dogs (8 per group) that had been inoculated 30 days prior with 50 third-stage D. immitis larvae were randomized to treatment on Day 0 with placebo or combination product, at the minimum dose of 24 µg/kg moxidectin, 2 mg/kg sarolaner and 5 mg/kg pyrantel (as pamoate salt). Study 2 also included groups treated with tablets containing moxidectin-alone (24 µg/kg) or sarolaner-alone (2 mg/kg). Efficacy was evaluated ~ 5 months after inoculation by adult heartworm counts at necropsy. In the field study, 410 dogs ≥ 8 weeks-old from 23 USA veterinary clinics were treated for 11 months with either combination product at 24–48 µg/kg moxidectin, 2–4 mg/kg sarolaner and 5–10 mg/kg pyrantel (n = 272) or Heartgard® Plus (ivermectin/pyrantel) at the label recommended dose rate (n = 138). Efficacy was evaluated on Day 330 using antigen and microfilaria testing to assess adult heartworm infection. Results In the laboratory studies, there were no heartworms recovered from any dog treated with the combination product or moxidectin alone and all dogs treated with placebo or sarolaner-alone were infected with 20–44 adult heartworms. In the field study, all dogs treated with the combination product tested negative for heartworm infection on Day 330, whereas two dogs treated with Heartgard® Plus tested positive. The Heartgard® Plus-treated dogs that tested heartworm positive were from the lower Mississippi River Valley region, where heartworm resistance has been confirmed to occur. The combination product was well tolerated in all studies. Conclusions In laboratory studies, no heartworms were recovered from dogs treated with a single dose of the novel combination product containing moxidectin, sarolaner and pyrantel. Additionally, in the field study no dog tested positive for adult heartworm infection when dosed with the combination product monthly for 11 months, while two dogs treated with Heartgard® Plus tested positive.

The localization of adult forms of Dirofilaria immitis in the heart provokes the development of the so-called “Caval syndrome” and lead to the development of severe cardiorespiratory disorders. The animals are in a severe general condition and surgical interventions must be précised as severity and duration. The purpose of this article is to revealed the methods and surgical techniques for removing these forms of the parasite from the heart as well as to describe the most common complications in the postoperative period.

Background Heartworm medications and many oral or topical flea and tick products are provided as monthly doses while a newer oral flea/tick product, fluralaner (BRAVECTO® Chew), is re-dosed at a 12-week interval. This study focused on whether there was a difference in the number of heartworm medication doses that were purchased in the 12-months follow-up period for dogs that receive either fluralaner or other flea/tick medications that are dosed monthly. Methods Clinic transaction records of heartworm medication purchases for over 200,000 dogs were examined to compare the purchase of heartworm preventative protection by dog owners that also receive flea and tick medications of differing efficacy durations. Results Annual purchases of heartworm medication for dogs by owners that receive a flea and tick medication dosed at 12-week intervals was incrementally higher than the number of doses purchased for dogs receiving monthly flea and tick medications. The average number of monthly doses per year was slightly over 7 months for both categories of product. The distribution of purchases of monthly doses was also similar between groups. Conclusions Dog owners who purchase a longer-acting flea and tick medication purchase as much heartworm medication annually for their dogs as dog owners who purchase monthly flea and tick medication. On average, dog owners who gave their dog fluralaner obtained significantly more months of heartworm preventative protection compared with dog owners who gave their dog a monthly flea and tick medication, although the biological significance of this increase in doses is very small.

Dirofilaria immitis (heartworm) is enzootic in many areas of the world and quite prevalent in southern European countries. Although dogs are the main host of the parasite, cats may also be infected, and the prevalence of feline dirofilariosis is associated with the respective prevalence of canine infection in any given area. The aim of the present study was to investigate the proportion of D. immitis infection among dogs and cats that were not under any kind of prophylactic treatment and were living in a heartworm enzootic area. In total, 180 stray animals (148 dogs and 32 cats) living in a shelter in Northern Greece were examined for heartworm infection by the Knott’s test and serology (antigen and in cats also antibody detection), and additionally echocardiography in the infected cats. Thirty seven (25%, CI: 18.7-32.5%) of the dogs and 3 (9.4%, CI: 3.2-24.2%) of cats were found to be positive, by at least one of the tests applied. In 2 of the infected cats the parasites were also detected by echocardiography. One of the positive cats died suddenly one year after diagnosis and at necropsy two decomposing D. immitis were found in the right ventricle and pulmonary artery. This is the first report of confirmed feline dirofilariosis in Greece. The detected proportion of infection in cats was 38% of the respective canine infection in the examined shelter. The results of the present study underline the high risk of infection of cats living in enzootic areas and the imperative character of preventive measures in such conditions.

Background A 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. Methods An 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. Results Positive 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. Conclusion These CAPC data show the rate of positive heartworm tests increasing significantly (P

Background Heartworm-associated respiratory disease (HARD) in cats is induced by the arrival and death of immature adult Dirofilaria immitis in the pulmonary system and is indistinguishable from mature adult heartworm infection. Methods A controlled, blind research study investigated the long-term (18 months post infection, PI) consequences of the inflammatory response associated with the death of immature adult heartworms in cats. Three groups of cats, 10 per group, were infected with 100 third-stage (L3) larvae by subcutaneous injection. Group A cats were treated with selamectin (Revolution®; Zoetis) per label directions at 28 days PI and once monthly for 17 months. Group B cats were treated orally with ivermectin (Ivomec®; Merial) at 150 μg/kg) at 70 days PI, then every 2 weeks for 15 months. Group C cats were untreated PI. At baseline (Day 0) and on Days 70, 110, 168, 240, 309, 380, and 505 PI, peripheral blood, serum, bronchial lavage, and thoracic radiographic images were collected. Results The selamectin-treated cats (Group A) and ivermectin-treated cats (Group B) were free of heartworms or heartworm fragments at necropsy. All cats became heartworm antibody positive at some time point in the study except for one cat in Group A. Only cats in Group C (all with adult heartworms) were heartworm antigen positive. The heartworm antibody titer for Group B was highest on Days 110 to 168 and then decreased over time and 50% were serologically antibody negative on Day 240. Eosinophilic bronchoalveolar lavage (BAL) cytology and peripheral eosinophilia were most pronounced on Day 110 in all cats. Randomly distributed myofibrocytes in the lungs of some Group A cats suggest that precardiac larval stages were affecting the lungs. Radiographs in Group B cats demonstrated partial resolution of the initial HARD reaction but chronic myofibrocyte proliferation was histologically evident 18 months after infection. Conclusion HARD was induced by immature adult worm infection with progressive improvement starting 6 to 8 months after infection but histologic lesions were evident in some cats 18 months after infection. The serologic antibody assay was negative in 50% of cats at 8 months and 100% of cats at 18 months post infection. Abnormal radiographic lung patterns continued in a subset of Group B cats for months after heartworm antibody serology and BAL cytology returned to normal.

Background The “susceptibility gap” in a dog diagnosed with adult heartworms has been defined as the period of time in which some Dirofilaria immitis stages are not susceptible to treatment with either macrocyclic lactones or melarsomine dihydrochloride. This was previously defined within the American Heartworm Society guidelines as a period of about 3 months “as per product labels.” It can be postulated, however, that a susceptibility gap does not exist with the combination of continued macrocyclic lactone therapy coupled with a three-dose melarsomine dihydrochloride protocol where the first intramuscular treatment is near the time of first diagnosis. Discussion Melarsomine dihydrochloride was originally also investigated as a “preventive” as well as a treatment for adult heartworm infection where it would be given to dogs by intramuscular injection every 4 months; therefore, there was early interest in its ability to kill younger worms. A single intramuscular injection of 2.5 mg melarsomine dihydrochloride/kg has an efficacy of 82.1% against 4-month-old worms. When it was given to dogs with older heartworms, 7 and 12 months of age, a single injection was only 55.6% and 51.7% effective, respectively. Thiacetarsamide has been shown to be 99.7% effective against 2-month-old heartworms and other work has shown that melarsomine dihydrochloride is 100% efficacious against these younger forms. With the development and US Food and Drug Administration (FDA) approval of spinosad + milbemycin oxime (Trifexis®, Elanco), milbemycin oxime + praziquantel (Interceptor® Plus, Novartis, now Elanco), and milbemycin oxime + lufenuron + praziquantel (Sentinel® Spectrum®, Novartis, now Virbac), it was shown that repeated treatments of dogs with milbemycin oxime also has efficacy against 3-month-old heartworms. Thus, no improvement in efficacy is expected with a delay in initiating therapy with both melarsomine dihydrochloride and macrocyclic lactones, even with the presence of younger heartworms. Starting treatment at diagnosis appears to be acceptable for maximal heartworm clearance based on published data. Delaying treatment has the disadvantage of allowing disease progression and continued heartworm growth. Conclusions The collective data that has been reviewed indicates that continued macrocyclic lactone administration with two additional injections of melarsomine dihydrochloride a month later will protect dogs against all heartworm stages, including those heartworms 2 months of age or younger at diagnosis, when both treatments are started upon diagnosis.

To evaluate whether heated serum allows for earlier detection of Dirofilaria immitis antigen, dogs with experimental D. immitis infections underwent weekly blood sampling to compare antigen results using both heated and unheated serum. One of two isolates (JYD-34 or Big Head™) were used to infect naive laboratory beagle dogs. Serum was collected from dogs weekly and divided into two aliquots, heated and unheated. The samples designated as heated were placed in a heat block at 104 °C for 10 min then centrifuged with collection of the resulting supernatant. Two commercial ELISAs, DiroCHEK® (Synbiotics Corporation, Zoetis) and PetChek® (IDEXX Laboratories, Inc.), were used to conduct D. immitis antigen testing on all serum samples. There was no statistical difference in the mean number of days from infection to positive D. immitis antigen status between the two commercial testing kits (DiroCHEK® versus PetChek®) with either heated or unheated serum. When unheated serum was utilized, very strong agreement between the two assays was demonstrated using Lin’s concordance correlation coefficient (R c = 0.98). However, when heated serum was compared, Lin’s concordance correlation coefficient was only R c = 0.64, showing a lesser agreement. There was a statistical difference in the mean number of days from infection to a positive test result for unheated serum when compared to mean days to positive status with heated serum. For DiroCHEK® the heated serum yielded a positive result 126.9 ± 18.9 days postinfection while the unheated serum yielded a positive result 162.6 ± 23.0 days postinfection; this was a significant 35.7 ± 32.2 days longer, on average, compared with heated serum. With PetChek® the heated serum yielded a positive result 131.5 ± 11.7 days postinfection while the unheated serum yielded a positive result 162.8 ± 23.8 days postinfection; this was a significant 31.3 ± 25.5 days longer, on average, compared with heated serum. The detection of D. immitis antigen earlier using heated serum was consistent for both heartworm isolates. Our results suggest heat treatment of serum may allow earlier detection of D. immitis antigen but with less consistency demonstrated across two testing platforms as compared with antigen detection using unheated serum.

Background The emergence of macrocyclic lactone resistance in canine heartworm poses a substantial threat to what is currently the only effective, FDA-approved available method of prevention. Further study of the biotypes is necessary to understand the mechanism of resistance and evaluate novel prevention options. Identifying cases of drug-resistant infection remains problematic, however, especially when poor compliance and insufficient testing are concerns. Furthermore, a definitive demonstration of resistance requires experimental infection and treatment, which is prohibitively costly. Methods With the aim of identifying likely cases of macrocyclic lactone-resistant heartworm and preventing their continued spread, we describe an algorithm for determining the likelihood of drug resistance and appropriate treatment strategies for each case. Results This algorithm relies on the microfilarial suppression test (MFST), which has been used previously as an efficient and discrete measure of suspected resistance. By standardizing this method in a format that is readily available to practitioners, it could become possible to preliminarily survey the emergence and spread of resistance. Conclusion Heartworm isolates identified through this method can be used in research to better understand macrocyclic lactone resistance so prevention strategies can be adapted.

Background The objective of heartworm treatment is to improve the clinical condition of the patient and to eliminate pre-cardiac, juvenile, and adult worm stages with minimal complications. Pulmonary thromboembolisms are an inevitable consequence of worm death and can result in severe pulmonary reactions and even death of the patient. To minimize these reactions, various treatment protocols involving melarsomine, the only adulticidal drug approved by the US Food and Drug Administrations (FDA), in conjunction with macrocyclic lactone heartworm preventives and glucocorticosteroids have been advocated. The discovery of the bacterial endosymbiont Wolbachia in Dirofilaria immitis has led to several experimental studies examining the effects of administering doxycycline to reduce or eliminate Wolbachia organism. These studies have shown a decrease in gross and microscopic pathology of pulmonary parenchyma in experimental heartworm infections pretreated with doxycycline before melarsomine administration. Methods Electronic medical records from a large veterinary practice in northeast Alabama were searched to identify dogs treated for heartworms with melarsomine from January 2005 through December 2012. The search was refined further to select for dogs that met the following criteria: 1) received two or three doses of ivermectin heartworm preventive prior to melarsomine injections, 2) received one injection of melarsomine followed by two injections 4 to 8 weeks later, and 3) were treated with prednisone following melarsomine injections. The dogs were then divided into those that also were treated with doxycycline 10 mg/kg BID for 4 weeks (Group A, n = 47) and those that did not receive doxycycline (Group B, n = 47). The medical notes of all 94 cases were then reviewed for comments concerning coughing, dyspnea, or hemoptysis in the history, physical exam template, or from telephone conversations with clients the week following each visit. Any dog that died within one year of treatment from either cardiovascular or pulmonary problems was noted. Results Dogs from Group A receiving doxycycline had fewer respiratory complications (6.52%) and heartworm disease-related deaths (0%) than Group B (19.14% and 4.25%, respectively). Conclusions Although there are not enough cases to indicate statistical significance, the results strongly suggest that including doxycycline into canine heartworm treatment protocols decreases post-treatment complications and mortality in naturally infected clinical cases.

Background Dirofilaria immitis infection occurs in dogs and cats, both of which species are clinically affected by mature adult infections. Cats are uniquely affected by immature-adult infections with an inflammatory pulmonary disease called Heartworm-Associated Respiratory Disease (HARD). D. immitis infection causes pulmonary parenchymal and vascular pathology in the dog and cat. Dogs develop pulmonary hypertension and cor pulmonale, whereas the development of pulmonary hypertension is rare in the cat. D. immitis infection in the dog causes alteration of the right ventricular (RV) extracellular matrix, including a decrease in myocardial collagen. In this study, the RV myocardial changes of cats infected with adult and immature-adult D. immitis were assessed. Methods The cardiopulmonary systems of six groups of SPF cats (n = 9-10 per group) were examined 8 or 18 months after infection with L3 D. immitis. Two groups were untreated and allowed to develop adult HW; two groups were treated with ivermectin starting 3 months post infection, thus allowing HARD but no mature adult heartworms; and two groups were treated with selamectin beginning 1 month post infection, preventing development of L5 or adult heartworms. A group of specific pathogen free (SPF) normal cats was utilized as a negative control (n = 12). Lung pathologic lesions were objectively assessed, and both RV and left ventricular (LV) weights were obtained to calculate an RV/LV ratio. Intramural RV myocardial collagen content was quantitatively assessed. Results RV/LV weight ratios were not different between groups. Negative control cats had significantly greater RV collagen content than all other affected groups (P = 0.032). Analysis of the RV/LV ratios and collagen content revealed no significant relationship (r = 0.03, P = 0.723, respectively). Collagen content had a modest, but significant, negative correlation, however, with both pulmonary vascular pathology (r = −0.25, P = 0.032) as well as the total pulmonary parenchymal and vascular pathology (r = −0.26, P = 0.025). Conclusions Cats infected with mature and immature D. immitis did not develop RV hypertrophy but did demonstrate loss of RV myocardial collagen content. The collagen loss was present at 8 and 18 months after infection in all infected cats. This loss of RV myocardial collagen was correlated with the severity of pulmonary parenchymal and vascular pathology.

Background Dirofilaria immitis is a worldwide parasite that is endemic in many parts of the United States. There are many commercial assays available for the detection of D. immitis antigen, one of which was modified and has reentered the market. Our objective was to compare the recently reintroduced Witness® Heartworm (HW) Antigen test Kit (Zoetis, Florham Park, NJ) and the SNAP® Heartworm RT (IDEXX Laboratories, Inc., Westbrook, ME) to the well-based ELISA DiroChek® Heartworm Antigen Test Kit (Zoetis, Florham Park, NJ). Methods Canine plasma samples were either received at the Auburn Diagnostic Parasitology Laboratory from veterinarians submitting samples for additional heartworm testing (n = 100) from 2008 to 2016 or purchased from purpose-bred beagles (n = 50, presumed negative) in 2016. Samples were categorized as “positive,” “borderline” or “negative” using our established spectrophotometric cutoff value with the DiroChek® assay when a sample was initially received and processed. Three commercially available heartworm antigen tests (DiroChek®, Witness® HW, and SNAP® RT) were utilized for simultaneous testing of the 150 samples in random order as per their package insert with the addition of spectrophotometric optical density (OD) readings of the DiroChek® assay. Any samples yielding discordant test results between assays were further evaluated by heat treatment of plasma and retesting. Chi-square tests for the equality of proportions were utilized for statistical analyses. Results Concordant results occurred in 140/150 (93.3%) samples. Discrepant results occurred in 10/150 samples tested (6.6%): 9/10 occurring in the borderline heartworm (HW) category and 1/10 occurring in the negative HW category. The sensitivity and specificity of each test compared to the DiroChek® read by spectrophotometer was similar to what has been reported previously (Witness®: sensitivity 97.0% [94.1–99.4%], specificity 96.4% [95.5–100.0%]; SNAP® RT: sensitivity 90.9% [78.0–100.0%], specificity 98.8% [96.0–100.0%]). There were significant differences detected when comparing the sensitivities of the SNAP® RT and the Witness® HW to the DiroChek® among the 150 total samples (p = 0.003) and the 50 “borderline” samples (p = 0.001). Conclusions In this study, the sensitivity of the Witness® HW was higher than the sensitivity of the SNAP® RT when compared with the DiroChek® test results prior to heat treatment of samples.

In heartworm disease, several biomarkers of cardiopulmonary injury and inflammatory activity have been studied during the recent years. D-dimer is a fibrin degradation product present after a clot is degraded, which has been reported to provide support for the diagnosis of pulmonary thromboembolism in heartworm disease. Furthermore, concentrations increment with increased disease severity and during the adulticide treatment. This increase in concentration has proved to be valuable. Cardiac biomarkers troponin I, myoglobin and NT-proBNP demonstrated presence of myocardial injury and heart failure, especially in chronic infections, which in some cases, slightly improve after the adulticide treatment. An acute phase response in dogs with Dirofilaria immitis, characterized by variations of acute phase proteins (APP), has been reported, indicating inflammatory processes that could contribute to disease progression. Among them, C-reactive protein (CRP) increases according to the severity of the disease; and a strong correlation between pulmonary hypertension and CRP has been observed. In cats, little work has been done to ascertain the utility of these biomarkers in feline heartworm; the only published study in D. immitis–seropositive cats reported significantly higher concentrations in positive APP serum amyloid A, haptoglobin and ceruloplasmin.

Background A controlled, blind research study was conducted to define the initial inflammatory response and lung damage associated with the death of immature adult Dirofilaria immitis in cats as compared with cats developing adult heartworm infections and cats on preventive medication. Methods Three groups of cats were utilized, 10 per group. All cats were infected with 100 third-stage (L3) larvae by subcutaneous injection. Group A cats were treated topically with selamectin (Revolution®; Zoetis) per label directions at 28 days post infection (PI) and once monthly for 8 months. Group B cats were treated orally with ivermectin (Ivomec®; Merial) at 150 μg/kg at 70 days PI, then every 2 weeks for 5 months. Group C cats were untreated PI. At baseline (Day 0) and on Days 70, 110, 168, and 240 PI, peripheral blood, serum, bronchial lavage, and thoracic radiographic images were collected on all cats. Upon completion of the study (Day 245), cats were euthanized and necropsies were conducted. Results Results were analyzed statistically between groups by ANOVA and by paired sample T testing for changes within the group over time. The selamectin-treated cats (Group A) did not develop radiographically evident changes throughout the study and were free of adult heartworms or worm fragments at necropsy. The heartworm life cycle was abbreviated with oral doses of ivermectin (Group B), shown by the absence of adult heartworms or worm fragments at necropsy. The early stage of immature adult worm in Group B cats, however, did induce severe pulmonary airway, interstitial, and arterial lung lesions, revealing that the abbreviated infection is a significant cause of respiratory pathology in cats. Cats in Groups B and C could not be differentiated based on radiographic changes, serologic antibody titers, complete blood count, or bronchoalveolar lavage cytology at any time point throughout the study. Eighty percent of cats in Group A and 100% of cats in Groups B and C became heartworm antibody positive at some time point post infection. Conclusions The clinical implications of this study are that cats that become infected with immature adult heartworms may not develop fully mature heartworms and are only transiently heartworm antibody positive, but do develop Heartworm-Associated Respiratory Disease (HARD).

Background Several reports have confirmed that macrocyclic lactone-resistant isolates of Dirofilaria immitis are circulating in the United States; however, the prevalence and potential impact of drug resistance is unknown. We wished to assess computer-aided measurements of motility as a method for rapidly assessing the resistance status of parasite isolates. Methods Blood containing microfilariae (MF) from two clinical cases with a high suspicion of resistance was fed to mosquitoes and the resultant L3 injected into dogs that were then treated with six doses of Heartgard® Plus (ivermectin + pyrantel; Merial Limited) at 30-day intervals. In both cases patent heartworm infections resulted despite the preventive treatment. Microfilariae isolated from these dogs and other isolates of known resistance status were exposed to varying concentrations of ivermectin in vitro and their motility assessed 24 h later using computer-processed high-definition video imaging. Results We produced two isolates, Yazoo-2013 and Metairie-2014, which established patent infections despite Heartgard® Plus treatments. Measurements of the motility of MF of these and other isolates (Missouri, MP3 and JYD-27) following exposure to varying concentrations of ivermectin did not distinguish between susceptible and resistant heartworm populations. There was some evidence that the method of MF isolation had an influence on the motility and drug susceptibility of the MF. Conclusions We confirmed that drug-resistant heartworms are circulating in the southern United States, but that motility measurements in the presence of ivermectin are not a reliable method for their detection. This implies that the drug does not kill the microfilariae via paralysis.

A controlled, blind study was conducted to define the initial inflammatory response and lung damage associated with the death of precardiac stages of Dirofilaria immitis in cats as compared to adult heartworm infections and normal cats. Three groups of six cats each were used: UU: uninfected untreated controls; PreS I: infected with 100 D. immitis L3 by subcutaneous injection and treated topically with selamectin 32 and 2 days pre-infection and once monthly for 8 months); IU: infected with 100 D. immitis L3 and left untreated. Peripheral blood, serum, bronchial lavage, and thoracic radiographic images were collected from all cats on Days 0, 70, 110, 168, and 240. CT images were acquired on Days 0, 110, and 240. Cats were euthanized, and necropsies were conducted on Day 240 to determine the presence of heartworms. Bronchial rings were collected for in vitro reactivity. Lung, heart, brain, kidney, and liver tissues were collected for histopathology. Results were compared for changes within each group. Pearson and Spearman correlations were performed for association between histologic, radiographic, serologic, hematologic and bronchoalveolar lavage (BAL) results. Infected cats treated with selamectin did not develop radiographically evident changes throughout the study, were heartworm antibody negative, and were free of adult heartworms and worm fragments at necropsy. Histologic lung scores and CT analysis were not significantly different between PreS I cats and UU controls. Subtle alveolar myofibrosis was noted in isolated areas of several PreS I cats and an eosinophilic BAL cytology was noted on Days 75 and 120. Bronchial ring reactivity was blunted in IU cats but was normal in PreS I and UU cats. The IU cats became antibody positive, and five cats developed adult heartworms. All cats with heartworms were antigen positive at one time point; but one cat was antibody positive, antigen negative, with viable adult females at necropsy. The CT revealed early involvement of all pulmonary arteries and a random pattern of parenchymal disease with severe lesions immediately adjacent to normal areas. Analysis of CT 3D reconstruction and Hounsfield units demonstrated lung disease consistent with restrictive pulmonary fibrosis with an interstitial infiltrate, absence of air trapping, and decrease in total lung volume in Group IU as compared to Groups UU and PreS I. The clinical implications of this study are that cats pretreated with selamectin 1 month before D. immitis L3 infection did not become serologically positive and did not develop pulmonary arterial hypertrophy and myofibrosis.

Aim: The aim was to study the pathomorphological changes through thoracic radiography and status of oxidative stress parameters in heartworm affected dogs in Odisha. Materials and Methods: A total of 16 dogs with clinically established diagnosis of dirofilariasis by wet blood smear and modified Knott’s test and equal numbers of dogs as control were included in this study. The present study was conducted in heartworm affected dogs to see the pathomorphological changes through thoracic radiography. Similarly, the evaluation was undertaken for observing any alterations in oxidative stress status in affected as well as non-affected, but healthy control dogs by adopting standard procedure. Results: Thoracic radiography revealed cardiac enlargement, round heart appearance suggestive of right ventricular hypertrophy, tortuous pulmonary artery and darkening of lungs. Alterations in oxidative stress indices showed a significant rise of lipid peroxidase activity, non-significant rise of superoxide dismutase and a significant although reverse trend for catalase levels in affected dogs in comparison to Dirofilaria negative control but apparently healthy dogs. Conclusions: Radiographic changes, as well as alterations in oxidative stress parameters, may not be diagnostic for heartworm infection, but useful for detecting heartworm disease, assessing severity and evaluating cardiopulmonary parenchyma changes and gives a fair idea about the degree of severity of the disease. It aids as contributing factors in disease pathogenesis.

Cats may be infected by heartworm, Dirofilaria immitis, through mosquito bites. They can develop severe heartworm disease when infective D. immitis larvae migrate and develop into adults in the pulmonary vasculature or other tissues. As there is no curative treatment for feline heartworm infection, the monthly administration of preventative treatment is recommended in endemic areas. Three controlled, blinded laboratory studies were conducted to evaluate the preventative efficacy of BROADLINE®, a novel combination of fipronil, (S)-methoprene, eprinomectin, and praziquantel against D. immitis in cats. In each study, 28 cats were inoculated with approximately 100 (studies 1 and 2) or 40 (study 3) infective third stage D. immitis larvae by subcutaneous injection, thirty days prior to treatment. The larvae were from recent field isolates from naturally infected dogs from three distinct geographic areas (two in the USA and one in Europe). In each study, the cats were allocated randomly to two study groups of 14 cats each. The control group remained untreated. On Day 0, each cat in the treated group received one topical application of the novel topical formulation, delivering the minimum intended dose of 0.5mg of eprinomectin per kilogram of body weight. At 6 months after infection, all cats were humanely euthanized and examined for adult D. immitis. Across all three studies, 28 (68%) of the 41 untreated cats harbored one or more heartworms, while 100% of the 42 treated cats remained free of heartworm infection, demonstrating the 100% preventive efficacy of BROADLINE® against D. immitis in cats. The treatment was well tolerated and no health abnormality was observed in any treated cat.

ResumenEl estudio de la parasitofauna de carnívoros silvestres es indispensable por su potencial uso como especies centinelas de la salud de los ecosistemas. Este estudio se llevó a cabo en la zona conurbada de Santiago de Querétaro, México, en donde se registró la presencia de filarias del nemátodo Dirofilaria immitis por medio de frotis sanguíneos en una zorra gris macho Urocyon cinereoargenteus, capturado en noviembre de 2013. Análisis posteriores señalaron la ausencia de nemátodos adultos en el corazón del cánido, pese al registro de las filarias en sangre. Se cuenta con registros previos de D. immitis en coyotes en Querétaro, pero se desconocía su presencia en este cánido silvestre.AbstractThe study of the parasite fauna of Mexican wild canids is relevant because of their potential as sentinel species for ecosystem health. The following study was conducted in a suburban area in Santiago de Querétaro, Mexico. We registered Dirofilaria immitis filariae in a gray fox male Urocyon cinereoargenteus blood sample on November of 2013. Further analysis showed there were not adult heartworms infecting the gray fox indicating that the animal was in the initial stages of infection. There are previous records of D. immitis in coyotes and stray dogs in Querétaro, but this nematode has yet to be registered in this host species.

Background Dog owners and veterinarians in small animal practices began to waive prevention of canine heartworm disease after heartworm infections seemed to have disappeared in Brazil. After 2013, infection rates rebounded, and an evaluation of the efficacy of chemoprophylactic drugs became necessary. Included in this re-evaluation was the efficacy of selamectin in client-owned dogs residing in a high infection-risk area. Methods The preventive efficacy of selamectin was evaluated by the topical application of selamectin to 24 client-owned dogs at the recommended rate (minimum of 6 mg/kg) by a veterinarian monthly for 36 months. Blood samples were collected before the first treatment and at the end of the study for testing to detect microfilariae by the modified Knott’s test and Dirofilaria immitis antigens using a commercial antigen test. Exposure to risk of heartworm infection was confirmed by the presence of infection in dogs living in low-income communities within a 2 km radius from the homes of dogs in the study. The dogs were managed according to routine practice by the owners within each household throughout the study. Results All dogs tested negative by both tests after receiving topical treatment with selamectin monthly for 36 months. Testing of 204 dogs from the communities confirmed the presence of heartworm in the area by detection of microfilariae or D. immitis antigen in 44 dogs (21.6 %). Conclusions Topical selamectin was 100 % effective for D. immitis prevention in 24 dogs that received monthly treatments by a veterinarian. Detection of heartworm infections in untreated dogs in the area suggests that clients need to be better informed regarding the prevalence of D. immitis and the importance of maintaining regular preventive treatments.

Heartworm disease in dogs can be severe and life threatening. Resistance to available heartworm preventives was considered among potential causes of increased reports of failed heartworm prevention in dogs. The objective of the present study was to compare the efficacy of four commercially available heartworm disease preventives against the JYD-34 strain of D. immitis. Forty laboratory-reared dogs approximately 6 months old were used. Each dog was infected with fifty, third-stage heartworm larvae on study day (SD) -30. On SD-1, the dogs were randomized to five groups of eight dogs each. On SD-0, dogs in groups 1–4 were treated as follows: Group 1: ivermectin/pyrantel pamoate chewable tablets; Group 2: milbemycin oxime/spinosad tablets; Group 3: selamectin topical solution; and Group 4: imidacloprid/moxidectin topical solution. Dogs in Group 5 were not treated and served as controls. The dogs were treated according to their current body weights and labelled dose banding for each product. Groups 1, 2, and 3 were retreated with their respective products and current body weights on SD 31 and 60. On SDs 124–126 the dogs were euthanized and necropsied for recovery of adult heartworms. Adult heartworms were recovered at necropsy from each of the dogs in the control group (13–32 worms/dog, geometric mean (GM) = 18.4 worms/dog). Adult heartworms and/or worm fragments were also recovered from each of the dogs treated with ivermectin/pyrantel pamoate, milbemycin oxime/spinosad or selamectin. Geometric means of worms recovered from dogs in each of these groups were 13.1, 8.8, and 13.1, resulting in efficacies compared to controls of 29.0, 52.2, and 28.8 %, respectively. All dogs in Group 4 (imidacloprid/moxidectin) were free of adult heartworms (100 % efficacy). The combination of imidacloprid/moxidectin was 100 % effective in this study in preventing development of JYD-34 laboratory strain of D. immitis in dogs following a single treatment, while three monthlytreatments of the three other commercial products provided less than 100 % efficacy. The high efficacy achieved with imidacloprid/moxidectin was likely due to the unique pharmacokinetic properties of the topical formulation delivering greater and sustained drug concentrations necessary to prevent development of D. immitis larvae.

This letter advises the imminent formation of the Companion Animal Parasites Council for the Tropics (CAPCT). The CAPCT consists of region-specific (e.g., Asia-Pacific, Latin America and Caribbean, Africa) experts comprising academics, veterinarians, parasitologists, physicians and allied industry partners that will work together to inform, guide and develop best-practice recommendations for the optimal diagnosis, treatment and control of companion animal parasites in the tropics, with the aim of protecting the health of pets and that of the public.

Human and animal parasitic filarial nematodes, which often are the cause of severe disease, harbor intracellular bacteria of the genus Wolbachia (Rickettsiaceae). It is thought that these bacteria play an important role in the pathogenesis and immune response to filarial infection. In order to determine the possible role of Wolbachia in heartworm disease, dogs naturally infected with Dirofilaria immitis were studied for specific antibody response to Wolbachia surface protein (WSP). Antibody subclasses were analyzed to determine immune response polarization. Dogs that died from heartworm disease were necropsied, and various organs were studied by immunohistochemistry to determine whether Wolbachia-derived molecules were present in tissue from infected dogs. Humoral response to the WSP was present in all infected dogs and appeared to be predominantly of the Th1-type. Several organs, including lung, liver, and kidney, contained positive-staining cells for WSP, confirming that the canine host does come into contact with Wolbachia-derived molecules.

Heartworm (Dirofilaria immitis) is a parasitic roundworm that causes a zoonotic disease known as dirofilariosis. Little is known about the role of wild carnivores serving as reservoirs in nature. Therefore, we examined 738 hearts and lungs of free ranging wild carnivores from Serbia to determine the presence of adult heartworms. During the period 2009–2013, the prevalence in golden jackals (Canis aureus) was 7.32 %, in red foxes (Vulpes vulpes) 1.55 %, in wolves (Canis lupus) 1.43 %, and in wild cats (Felis silvestris) 7.69 %. No adult heartworm specimens were found in beech martens (Martes foina), stone martens (Martes martes), European polecats (Mustela putorius), badgers (Meles meles) or otter (Lutra lutra). The highest recorded prevalence was in 2013 (7.30 %) and the lowest in 2012 (1.6 %). In jackals, the prevalence was higher in males (10 %) than in females (4.06 %), while in foxes the prevalence was 1.75 % in males and 1.26 % in females. The most infected host was a wolf in which 37 adult specimens were found. Because of the potentially significant role in the life cycle of D. immitis, populations of wild carnivores in Europe should be further examined and tested for heartworm infections.

A controlled, blind research study was conducted to define the innate response of lungs in specific pathogen free (SPF) cats to intravenous ( n = 10) or subcutaneous ( n = 4) administration of homogenate of adult Dirofilaria immitis from donor dogs compared with lung response in control cats ( n = 6). There was no difference in cats that received heartworm homogenate IV for 18 days from donor dogs treated with doxycycline for 1 month compared with cats given heartworm homogenate from untreated donor dogs. Cats did not develop clinical signs, and no radiographic changes were noted. Cats given SC heartworm homogenate at lower concentration than IV groups did not develop histologic changes. Cats that received IV heartworm homogenate for 18 days developed mild interstitial and peribronchial myofibrocyte proliferation and smooth muscle proliferation of the pulmonary arteries. Bronchial ring contractility in vitro was blunted in the IV homogenate cats to the agonists acetylcholine and 5-hydroxytryptamine. Cats in the SC group had increased sensitivity to histamine at high concentrations but normal contractility and relaxation responses to other agonists. No increase in mast cells was noted in lung tissues of cats given homogenate. In the absence of bronchial wall remodeling, cats given IV homogenate had blunted responses to bronchial constriction, but normal relaxation to nitroprusside and substance P and increased sensitivity to histamine. In the absence adult heartworms, the homogenate of adult heartworms in the circulation of SPF cats induced a direct effect on lung parenchyma and altered bronchial ring reactivity.

Dirofilariosis is a severe vector-borne emergent disease that is spreading worldwide and becoming a serious threat to human and veterinary public health. Portugal, a Mediterranean country, has favorable climate conditions for mosquito development and survival. At present, accurate data on the prevalence and epidemiological pattern of dirofilariosis in Portugal is scarce and outdated. To study these trends, a project was developed to assess the current prevalence of Dirofilaria immitis infection in shelter dogs as well as the prevalence of Dirofilaria species present in three coastal regions in central Portugal: Coimbra, Santarém, and Setúbal. Blood samples were collected from 696 shelter dogs during three consecutive years: 2011, 2012, and 2013. A rapid immunomigration technique was performed to detect female D. immitis antigens. Concurrently, to detect and identify circulating microfilariae, a modified Knott's technique and acid phosphatase histochemical staining were also performed. Of the 696 dogs sampled, 105 were positive for D. immitis, with an overall prevalence of 15.1%. Forty of the 105 dogs were antigen negative but were positive for D. immitis microfilariae. Three animals were co-infected with D. immitis and Acanthocheilonema dracunculoides, and there was also one dog infected only with A. dracunculoides, all confirmed by polymerase chain reaction. The highest prevalence of canine heartworm during the 3-y period was in Setúbal (24.8%), followed by Coimbra (13.8%), and Santarém (13.2%), with significant inter-district differences. Our results demonstrate a higher prevalence of dirofilariosis compared with findings of previous studies and show an increasing rate of infection in the southern areas of Portugal attributed, at least in part, to bioclimatic and ecological factors. The present study updates the epidemiological situation and correlates the risk of dirofilariosis transmission within each region. These findings are highly relevant to both human and veterinary public health, contributing to the general awareness of pet owners and veterinarian practitioners and reinforcing the need for effective control measures against vectors and preventive therapy in companion animals.

Six hundred and eleven random-source dogs (338 male, 273 female) one year of age or older, from six sections of the city of Recife, Pernambuco, were examined antemortem for circulating microfilariae Dirofilaria immitis and Dipetalonema reconditum adult heartworm (D. immitis) antigen, and examined postmortem for adult heartworms. The prevalence of heartworm infection was 2.3% (14/611), as determined by necropsy for adult worms, and 1% (6/611) had circulating microfilariae of D. immitis; thus, 57.1% of the heartworm-infected dogs had occult infections. The results of serological testing indicated that 1.3% (8/611) of the dogs were positive for adult heartworm antigen. A total of 42 (6.9%) of the dogs had microfilariae of D. reconditum; 40 of these had only D. reconditum and two additional dogs had microfilariae of both species, D. immitis and D. reconditum.

Dirofilaria immitis (heartworm) infections affect domestic dogs, cats, and various wild mammals with increasing incidence in temperate and tropical areas. More sensitive antibody detection methodologies are required to diagnose asymptomatic dirofilariasis with low worm burdens. Applying current transcriptomic technologies would be useful to discover potential diagnostic markers for D. immitis infection. A filarial homologue of the mammalian translationally controlled tumor protein (TCTP) was initially identified by screening the assembled transcriptome of D. immitis (DiTCTP). A BLAST analysis suggested that the DiTCTP gene shared the highest similarity with TCTP from Loa loa at protein level (97%). A histidine-tagged recombinant DiTCTP protein (rDiTCTP) of 40 kDa expressed in Escherichia coli BL21 (DE3) showed immunoreactivity with serum from a dog experimentally infected with heartworms. Localization studies illustrated the ubiquitous presence of rDiTCTP protein in the lateral hypodermal chords, dorsal hypodermal chord, muscle, intestine, and uterus in female adult worms. Further studies on D. immitis-derived TCTP are warranted to assess whether this filarial protein could be used for a diagnostic purpose.

Objective: This study was designed to assess the prevalence and the associated risk factors (e.g., sex, age, breed, management system and climate) of Dirofilaria immitis in dogs in Makurdi metropolis in Nigeria. Materials and methods: Prevalence study of canine heartworm disease in dogs was conducted over a period of six months covering five localities of Makurdi metropolis in Benue State, Nigeria. A total of 186 blood samples were collected from apparently healthy and sick dogs, and the samples were examined for the presence of microfilaria between September 2015 and February 2016. Three methods (wet mount, Buffy coat and modified Knott's techniques) were used for the examination of the samples. The Packed Cell Volume (PCV) and complete blood count for each sample were also determined. Results: Out of the 186 dogs, 4 (2.15%) were found to be positive for the presence of microfilaria. Out of the 4 positive cases, 3 (1.61%) were microfilaria and 1 (0.54%) was unidentified motile parasite. A total of 104 females were examined and only 1 (0.96%) was positive, while 3 (3.66%) males out of 82 examined were positive. Out of 141 older dogs examined, 4 (2.84%) were positive. Hematology of the positive dogs revealed mild anemia and moderate thrombocytopenia with Mean+/-SD of 34.8+/-15.30% and 108+/-60.81x109/L, respectively. Conclusion: The study confirms Knott's technique to be the most sensitive in the diagnosis of dirofilariasis in dogs using parasitological techniques. The findings confirm the occurrence of D. immitis in dogs in Makurdi with low prevalence and that the general public are at high risk of spreading infection from the dogs. Infection is more in male and adult dogs. This work can assist in planning appropriate strategies for controlling and prevention of D. immitis infection in Nigeria. [J Adv Vet Anim Res 2016; 3(4.000): 338-344]

Carval syndrome is a severe heartworm infection where the worms have migrated to the right atrium and vena cava; this condition is associated with a myriad of clinical signs. Several non-surgical and interventional methods are currently used for mechanical worm removal. However, the success rate and complications related to these methods are heavily dependent on methodology and retrieval devices used. In this study, we developed a catheter-guided heartworm removal method using a retrieval basket that can easily access pulmonary arteries and increase the number of worms removed per procedure. With this technique, we successfully treated four dogs with caval syndrome.

Canine heart worm disease is often life-threatening due to its various complications, including right side heart failure, caval syndrome and pulmonary eosinophilic granulomatosis. Several preventive medications and melarsomine have been developed and they are very effective to control heartworm infestation. However, in a case of severe infestation, melarsomine therapy often results in an unfavorable outcome because of the severe immune reaction caused by rapid killing of the adult worm. Surgical removal and an interventional method using flexible alligator forceps have been well described in the literature. Despite the usefulness of mechanical removal using flexible alligator forceps, the methodology still needs to be upgraded for increasing the applicability for treating dogs with severe infestation. We describe herein a newly developed percutaneous removal method for heartworms and this was successfully applied to 4 dogs with severe heartworm infestation. The follow-up studies also showed favorable outcomes with no complications.

3 pages.-- PMID: 7760485 [PubMed]., We determined the prevalence of Dirofilaria immitis (Nematoda, Filariidae) among 433 red foxes (Vulpes vulpes) in northeastern Spain, between 1990 and 1992. Forty-six (11%) of 433 foxes were infected; the intensity ranged from 1 to 36 (mean ± SE; 4.39 ± 0.92) nematodes per host. The prevalence of D. immitis was higher in foxes inhabiting riparian zones of the study area. This population has a very high juvenile/adult ratio. Heartworm prevalences did not differ among host sex, weight, or fat condition categories., The fieldwork was partially financed by the Government of Aragón, Programme 533.1 of Fauna Conservation.

La dirofilariosis canina, conocida también como “gusano del corazón”, es una parasitosis ocasionada por el nemátode Dirofilaria immitis. Se localiza en el corazón de los caninos ocasionando una enfermedad cardiopulmonar que repercute en el hígado y riñones. Los hospederos intermediarios son los mosquitos hembras del género Culex, Aedes y Anopheles. El objetivo del estudio fue evaluar la prevalencia de este parásito en los caninos del distrito de San Juan de Lurigancho (ribereño al río Rímac). Se tomaron al azar muestras de sangre de 107 perros, aparentemente sanos, mayores de 6 meses, sin distinción de raza o sexo, entre mayo a junio del 2002. Se utilizó para su diagnóstico la prueba enzimática de ELISA (Dirochek. Synbiotic®). Se hallaron 6 animales positivos, los cuales representan una frecuencia de 5.5±4.7%. No se encontraron diferencias debidas a sexo o tiempo de permanencia en el domicilio. Se concluye que la frecuencia de dirofilariosis en los caninos de este distrito fue moderada y se mantiene similar con relación a los diagnósticos inmunoenzimáticos previos realizados en diversos distritos de Lima Metropolitana., Canid filariasis, known as Heartworm disease, is caused by a nematode named Dirofilaria immitis, which is located in the canid´s heart producing a cardiopulmonary disease and affecting others organs like liver and kidneys. Culex, Aedes and Anopheles female flies are the intermediary hosts. The objective of this study was to determine the prevalence of D. immitis in the San Juan de Lurigancho district, close to the Rímac river. Blood samples were taken from 107 dogs in apparently good health condition from May to June 2002. Animals were randomly chosen without regard of breed and sex, and were older than 6 months old. An enzimatic commercial ELISA test (Dirochek, Synbiotic®) was used. Six positive dogs were detected, representing a frequency of 5.5±4.7% with a 95% confidence interval. No differences due to sex and environment exposition were found. In conclusion, the frequency of dirofilariasis in canids of this district is low and similar to previous studies conducted in various districts of Lima.