Comparison of heartworm antigen test kit performance in dogs having low heartworm burdens

Sensitivity and specificity of four in-clinic heartworm antigen test kits, AbboScreen (Abbott Laboratories), Snap PF (IDEXX Laboratories), Solo Step (HESKA Corporation), Witness (Synbiotics Corporation) and two heartworm antigen microwell plate assays, DiroCHEK (Synbiotics) and PetChek PF (IDEXX) were compared in a blinded study using serum or plasma drawn from 237 random source dogs, including 140 with necropsy-confirmed, low worm burden infections (minimum 1 worm, maximum 10, mean 2.3, median 3) and 97 confirmed heartworm-free at necropsy. In general, microwell format tests were more sensitive than membrane format tests and tests using ELISA technology were more sensitive than tests using lateral flow immunochromatographic technology. Percent sensitivity and specificity, respectively, were PetChek PF 76 and 97, DiroCHEK 71 and 94, SNAP PF 67 and 98, Solo Step 60 and 98, and AbboScreen 52 and 96. The Witness test protocol was changed by the manufacturer midway through the study, and the newer version of this test kit arrived containing a package insert alerting the user to a change in procedure, which purportedly resulted in improved sensitivity. PetChek was significantly more sensitive than all other tests except DiroCHEK and the new version of Witness. DiroCHEK was significantly more sensitive than all tests except PetCheck, SNAP and the new version of Witness. Snap was more sensitive than AbboScreen and the old version of Witness. Differences in specificity were not significant (P>0.05).     

Comparative Evaluation of 2 In-Clinic Assays for Vector-Borne Disease Testing in Dogs

Vector-borne agents comprise medically important infections affecting dogs throughout much of the world. Sensitive detection of antibodies directed at tick-borne disease-causing organisms in dogs is diagnostically important for veterinarians, pets and their owners, and epidemiologically important for public health surveillance. The SNAP 4Dx Plus Test (IDEXX Laboratories, Inc., Westbrook, ME) identifies antibodies to or infection with multiple tick-borne pathogens and canine heartworm antigen in a single assay. Recently, VetScan FLEX4 Rapid Test (Abaxis, Inc., Union City, CA) was launched as a new assay to detect tick-borne pathogen antibodies and heartworm antigen. In the present study, we evaluated the comparative performance of SNAP 4Dx Plus (SNAP) and FLEX4 Rapid Test (FLEX4) using samples selected based on geographic distributions for canine vector borne diseases, including Borrelia burgdorferi (n = 105), Anaplasma phagocytophilum (160), Anaplasma platys (115), Ehrlichia canis (154), Ehrlichia ewingii (163), Ehrlichia chaffeensis (151) and Dirofilaria immitis (105). Canine vector borne diseases infection status was established for each sample by a combination of reference methods that included necropsy (D. immitis, heartworm disease), Western immunoblotting (B. burgdorferi), immunofluorescence assays (A. phagocytophilum and E. canis) and species-specific ELISAs (A. platys, E. canis, E. ewingii and E. chaffeensis). For comparisons among the 2 assays, samples were evaluated per the manufacturers’ instructions for each test kit.By testing each same sample set compared to the defined reference results, sensitivities differed substantially between SNAP and FLEX4, at 95.5 vs. 40.9%, respectively for B. burgdorferi, 97.1% vs. 61.4% for E. canis, 98.2% vs. 59.3% for E. ewingii, 64.3% vs. 35.7% for E. chaffeensis, 84.5% vs. 12.7% for A. phagocytophilum, 83.3% vs. 33.3% for A. platys, and 94.1% vs. 88.2% for D. immitis. Specificities for both rapid assay tests ranged from 98% to 100%.Based upon the comparative results derived from this study, the SNAP test was more sensitive than the FLEX4 test for detection of antibodies to all tick-borne pathogens and heartworm disease (Dirofilaria immitis) antigen in dogs.     

Validation of an in-clinic enzyme-linked immunosorbent assay kit for diagnosis of Borrelia burgdorferi infection in horses.

Confirmation of Borrelia burgdorferi infection in horses has required enzyme-linked immunosorbent assay (ELISA) or Western blot tests performed by reference laboratories. An in-clinic C6 ELISA SNAP kit has been marketed for dogs. This canine kit was evaluated for horses using serum from experimentally infected ponies. Serum samples originated from 2 previous studies. In the first study, 7 ponies were exposed to B. burgdorferi-infected ticks; 4 ponies served as uninfected controls. Serum samples were obtained bimonthly for 9 months. In the second study, 16 ponies were exposed to B. burgdorferi-infected ticks. After confirmation of infection by skin culture, polymerase chain reaction (PCR), and serology, the ponies were allocated to 4 groups that received tetracycline, doxycycline, ceftiofur, or no treatment. Serum samples were obtained monthly, both before and after antibiotic treatments, for 11 months. For the current study, selected samples (n = 220) from both studies were tested with IDEXX SNAP Heartworm Ab/Borrelia burgdorferi Ab/Ehrlichia canis Ab Test Kits. Tested samples included samples taken before infection, from various times postinfection, and after antibiotic treatments. Results from confirmed positive or negative samples were used to determine sensitivity and specificity of the assay. Results indicate that the test kits have fair sensitivity (63%) and very high specificity (100%) for horses recently infected with B. burgdorferi. Validation of this test provides equine practitioners with an inexpensive, in-clinic method to confirm infection, although its moderate sensitivity may result in a moderate chance of a false negative test.     

Comparison of results of three commercial heartworm antigen test kits in dogs with low heartworm burdens

OBJECTIVE: To compare results of 3 commercial heartworm antigen test kits performed on serum samples from dogs infected with low numbers of adult female heartworms. DESIGN: Blinded laboratory evaluation. Sample Population-Serum samples from dogs (n = 208) proven at necropsy to be infected with 1 to 4 adult female heartworms and from dogs (32) without heartworms. PROCEDURE: Samples were sequentially tested with each test kit, following the manufacturers' instructions, by licensed veterinary technicians in private practice who were not aware of infection status of the dogs. The order of test kit evaluations was randomly chosen. For each test kit, sensitivity, specificity, accuracy, positive predictive value, and negative predictive value were evaluated. RESULTS: All tests yielded some false-negative results, and there were significant differences among tests in regard to ability to detect low heartworm burdens. Sensitivity of the test kits ranged from 78 to 84%. For all test kits, sensitivity increased as number of female heartworms increased. All 3 test kits had high specificity (97%). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that sensitivity of the 3 commercially available heartworm antigen test kits ranged from 78 to 84% when used to test serum samples from dogs with low heartworm burdens, and that sensitivity varied among test kits. For all 3 test kits, specificity was 97%. All 3 test kits yielded false-positive and false-negative results for some dogs with low heartworm burdens.     

Dirofilaria immitis prevalence in a canine population in the Samana Peninsula (Dominican Republic) - June 2001

Dirofilaria immitis, the etiological agent for heartworm disease, is a zoonotic, vector-borne parasite. The dog is the most common natural host. Heartworm disease is present in all US states, as well as in South and Central America, Asia, Europe, Africa and Australia [. Prevalence of feline heartworm disease - a global review. In: Proceedings of the Heartworm Symposium'95, Auburn, Alabama, USA, 31 March-2 April, 1995. American Heartworm Society, Batavia, USA, pp. 79-86]. According to Menda [Menda, J.A., 1989. Transplacental migration of Dirofilaria immitus microfilariae. Companion Anim. Pract. 19 (6-7), 18-20], 18% of the canine population in the city of Santo Domingo tested positive for heartworm using the Filar Assay Kit. In the year 2000, clinical veterinarians from the Samana Peninsula in the Dominican Republic stated that the incidence of heartworm was minimal (Goodman, personal communication). In this small epidemiological study performed during the summer of 2001, 104 dogs from the Samana Peninsula were tested for heartworm antigen with the WITNESS HW Test and a wet mount evaluation for microfilaria under light microscopy. It was found that 18.2% of the canine population tested positive according to either one or both of the diagnostic methods. It would be advisable to apply prevention measures to control Dirofilaria immitis infection in the canine population of the Samana Peninsula in the Dominican Republic.     

The validity of some haematological and elisa methods for the diagnosis of canine heartworm disease

Examinations for heartworm (Dirofilaria immitis) were performed on 175 impounded dogs from a hyperendemic area of the Po Valley (Italy). Each blood sample was used wiht five haematological diagnostic methods (filtration, direct smear, modified Knott, clotted blood and capillary tube) and three commercial ELISA kits (PetChek, Diasystems, Uni-Tec). The results were compared with the true infection status obtained from post-mortem examination of the heart, pulmonary arteries, thoracic venae cavae and lungs. The prevalence of the infection by adult worms at necropsy was 63%. The sensitivity of the tests ranged from 60% (capillary tube) to 81% (Diasystems) and the specificity from 88% (filtration) to 98% (PetChek). The results of all the tests differed significantly (p<0.01) from those obtained at necropsy. The sensitivity of the tests was also assessed with respect to the differing numbers of worms in the hosts. A positive correlation between the worm burden and the sensitivity was observed in all the tests. It is apparent that the ELISA methods were better able to detect cases with a low number of worms than the haematological tests.Abbreviations AC accuracy - CB clotted blood - CT capillary tube - DI Diasystems - DS direct smear - ELISA enzyme-linked immunosorbent assay - FI filtration - MK modified Knott - NPV negative predictive value - PE PetChek - PPV positive predictive value - SE sensitivity - SP specificity - UN Uni-Tec     

Field efficacy and safety of a combination of moxidectin and imidacloprid for the prevention of feline heartworm (Dirofilaria immitis) infection

Throughout the end of March to beginning of May 2006, 212 owned cats and 608 owned dogs from a heavy endemic area for canine heartworm (HW) disease in northern Italy have been examined to assess HW infection prevalence. Both cats and dogs were clinically examined and blood samples were taken from each animal to be examined for HW antibody (Ab). Ab-positive cats were further examined for circulating microfilariae, HW antigens (Ag) and by echocardiography (ECHO) to assess the presence of adult worms. Dogs were clinically examined and blood samples taken from each animal were examined for circulating microfilariae and for HW Ag. Ten cats (4.7%) were found Ab positive. Of these, 6 cats were Ag positive (2.6%) and in 4 (1.8%) the worms were visualized by ECHO. HW prevalence in dogs was 36% (221/608). One hundred and seventy-six (29%) were both microfilaraemic and Ag positive, 40 (7%) had occult infections (no circulating microfilariae) and 7 (1%) were microfilaraemic but Ag negative. Upon owners’ consent, 132 cats (including cats Ab and/or Ag and ECHO positive) were prophylactically treated against HW disease with an imidacloprid/moxidectin spot-on combination (10% imidacloprid/1% moxidectin) monthly administered for 6 months. Cats were re-examined for HW infection in November, 1 month after the last drug administration, and in May–June 2007, 7–8 months after the last treatment. All 122 cats found HW negative before treatment, were found negative at the two examinations at the end of study. The 4 cats Ab positive, 2 cats Ab and Ag positive and 1 Ab, Ag and ECHO positive at the beginning of treatment were found negative. Throughout the treatment, transitory hypersalivation and generic signs of annoyance were reported by owners in 6 cats (4.5%). All signs regressed spontaneously.     

Dirofilaria immitis: worm burden and pulmonary artery proliferation in dogs from Michigan (United States)

Despite the ability to prevent heartworm disease, infection with Dirofilaria immitis continues to be a major problem for domestic dogs. To determine worm burden in heartworm-positive dogs from three county animal shelters in the state of Michigan in the United States and to assess the relationship between gross intimal proliferation and worm burden, necropsy was done on 176 heartworm-positive dogs. Adult heartworms were found in the heart and pulmonary artery of 170 of the 176 (96.6%) dogs examined. Mean worm burden was 14 +/- 13 (range 0-85). Fifty-nine percent of dogs had < or =10 heartworms. In contrast, 52% of dogs in a published report from the southern US (Florida) had worm burdens >10 [C.H. Courtney, Q.Y. Zeng, The structure of heartworm populations in dogs and cats in Florida, in: Proceedings of the American Heartworm Symposium, 1989]. These data suggest that mean worm burden in northern areas may be < or = that in warmer areas. Also, since diagnostic tests are less sensitive with lower worm burdens, diagnosis of heartworm infection in Michigan and other surrounding more northern states may be a greater challenge than in areas with higher worm burdens.     

Prevalence and distribution of Dirofilaria immitis in domestic dogs from Ankara and vicinity in Turkey

A total of 280 dogs in Ankara and vicinity were examined from November 2000 through December 2001 by the PetChek antigen ELISA and polycarbonate filter-acid phosphatase histochemical staining techniques for Dirofilaria immitis infection. Dirofilaria immitis (heartworm) infection was found in 26 dogs (9.3%), all of which were kept always outdoors and non prophylacted by their owners. The distribution of heartworm infected dogs was not uniform. Infection with D. immitis was correlated with differences in age and sex. But, no significant association was found.     

An investigation of the role of immunologic factors in anemia associated with canine heartworm disease

Direct Coombs' antiglobulin tests were performed on 80 dogs with patent Dirofilaria immitis infection and 170 dogs negative for microfilaria of D. immitis. Presence or absence of anemia was determined by hematocrit in 55 of the heartworm negative dogs and 68 of the dogs with heartworm disease. Heartworm infected dogs showed a higher incidence (37%) of anemia than noninfected dogs (14.5%). Anemia was most prevalent in two groups of dogs with heartworm infection, one group showing vena caval syndrome (91%) and the other occult dirofilariasis (62.5%). These latter two groups of dogs also showed a significantly higher number of positive Coombs' reactions at 37 degrees C than other dogs with heartworm disease and the noninfected dogs. The number of positive Coombs' reactions at 4 degrees C among the total of 80 dogs with heartworm infection was significantly higher than that for dogs without heartworm disease. However, there was no positive correlation between anemia and the outcome of the Coombs' test at either temperature. These findings do not suggest that immunologic factors play a major role in the pathogenesis of anemia in dogs with heartworm disease.     

Serological update and molecular characterization of Dirofilaria immitis in dogs,South Korea

Eighty-one dogs in the Chungnam province were tested for heartworm (Dirofilaria immitis) infection by ELISA (SNAP test, IDEXX Laboratories, Maine, USA). Seventeen (20.9%) of the 81 samples were found to be positive and further analyzed by 16S rRNA sequencing. In this study, all dogs tested lived outdoors. Using the χ² test and Fisher’s exact test, no significant differences in the prevalence of dirofilariasis were observed among different gender and age groups, although the prevalence of this disease among dogs 2–4 yrs of age remains highest. Sequence analysis revealed that the species prevalent in Chungnam province were genetically distinct from the type strain of D. immitis based on the nucleotide deletion found at position nt 276 (cytosine) and nucleotide substitution at position 428 (G to A) of the partial 16S rRNA sequence of the type strain. Furthermore, phylogenetic analysis suggests at least 2 groups of D. immitis circulating in the Chungnam area between the year 2007 and 2008.     

Comparison of four serotests for the detection of Dirofilaria immitis infection in dogs

Three hundred two dogs were tested with 4 serotests for heartworm antigen (AG) or antibody (AB) and with the Knott test. The 4 serotests evaluated were an enzyme-linked immunosorbent assay (ELISA) for adult heartworm-specific AB (AB-ELISA), a quantitative, indirect immunofluorescent assay (IFA) for adult heartworm-specific AB (AB-IFA), an IFA test for microfilaria (MF)-specific AB (MF-IFA), and an ELISA for adult heartworm AG (AG-ELISA). The presence of heartworms was ascertained in all dogs by necropsy examination. Of 302 dogs, 20 (6.6%) had heartworms in the heart at necropsy. Of infected dogs, 9 (45%) had occult infections. Test sensitivities were 75%, 95%, 70%, and 75% for the AB-ELISA, AB-IFA, MF-IFA, and AG-ELISA, respectively. Test specificities were 85% (AB-ELISA), 77% (AB-IFA), 87% (MF-IFA), and 99% (AG-ELISA). The best agreement between serotest results and necropsy findings was obtained with the AG-ELISA (97%). The 4 serotests detected 86% (AB-ELISA), 100% (AB-IFA), 67% (MF-IFA), and 78% (AG-ELISA) of the dogs with occult heartworm infection. A significant (P less than 0.05) association between intestinal parasitism and positive heartworm test results was found with only AB-IFA. Seemingly, the Knott test, or some other concentration method for detecting circulating MF should be the first heartworm test performed. If the examination for MF is negative, the dog has clinical signs, and radiographic findings are suggestive of occult heartworm infection, then a serotest for adult heartworm AG is recommended.     

Epidemiological aspects of filariosis in dogs on the coast of Paraná state, Brazil: with emphasis on Dirofilaria immitis

The present study determined the prevalence and geographical distribution of Dirofilaria immitis and other filariae, from dogs in littoral areas of Paraná state, in Brazil. This survey spanned eight months, between 1998 and 1999, and was also designed to compare the efficacy of different tests for diagnosis of heartworm infection in that area. Blood samples were collected from 256 native-owned dogs distributed along the Paraná coastal area. Five diagnostic procedures were used: direct smear examination, the Knott's modified test, filtration assay, and two heartworm antigen detection kits. A follow-up imaging exam was performed to support the heartworm diagnosis. The imaging diagnosis included radiographic and ultrasonographic exams of six dogs that had positive results for the heartworm antigen detection kits, but showed different microfilarial burdens. The presence and severity of radiographic and ultrasonographic signs were compared with the results obtained in microfilariae detection and antigen tests. Diagnostic parasitology results indicated that 31.25% of the dogs were microfilaremic. Three different microfilariae were recovered: D. immitis, Dipetalonema reconditum, and the third (mf3) was not identified. D. reconditum was the species with the highest prevalence: 22.6%. In general, D. immitis prevalence was 5.47% (28.57% occult infections), but it varied along the coast and the range was from 0 to 20%. No correlation could be established between the overall scores for microfilarial counts (small or large numbers) and the severity of radiographic results or the likelihood of detecting filariae in the pulmonary artery using echocardiography. The finding of a different type of microfilaria (mf) suggested the existence of a third species in Paraná state, whose prevalence was 4.68%. These results show that to obtain a reliable diagnosis of heartworm infection, antigen detection kits are indicated. Knott's test or filtration should be performed to confirm microfilaremia and not for diagnosis of heartworm infection. Imaging tests support parasitology exams and add more about severity of infection. The northern areas, specially Guaraque?aba and Ilha das Pe?as, presented the highest number of heartworm-infected dogs.     

Prevalence of heartworm infection in healthy cats in the lower peninsula of Michigan

OBJECTIVE: To determine prevalence of heartworm infection among healthy, client-owned cats in the lower peninsula of Michigan. DESIGN: Cross-sectional prevalence study. ANIMALS: 1,348 healthy cats examined at private veterinary practices throughout the lower peninsula of Michigan. PROCEDURE: Sera were tested by use of an ELISA-based antigen test kit to determine infection and 2 commercially available antibody detection kits to determine exposure. A questionnaire was used to collect data to assess risk factors associated with infection. RESULTS: 25 cats had positive results for heartworm antigen, yielding an observed prevalence of 1.9%. Neither antibody test was reliable or provided reproducible results, and neither yielded positive results for more than 20% of the antigen-positive heartworm-infected cats. Multivariate regression indicated that cats from southeastern Michigan and cats > or = 2 years old had a higher risk of infection. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that most (80%) heartworm-infected cats in the lower peninsula of Michigan were from the southeastern part of the state, a pattern that closely paralleled the prevalence of heartworm infection in dogs. Therefore, knowledge of the regional prevalence of heartworm infection in dogs may be useful in assessing the risk of infection in cats. Results also suggested that currently available in-clinic heartworm antibody detection kits have limited utility in the diagnosis of heartworm infection in cats.     

Can heartworm prevalence in dogs be used as provisional data for assessing the prevalence of the infection in cats?

Cats are considered a susceptible host for Dirofilaria immitis; however, increased host resistance is reflected by relatively low adult worm burdens in natural and experimental infections; the prolonged prepatent period (8 months); the low level and short duration of microfilaremia; and the short life span of adult worms (2-3 years). From April to September 2006, 212 cats and 608 dogs, all exposed for at least one transmission season, were screened for D. immitis infection in a multi-center study in the Po River Valley in northern Italy. Cats were initially evaluated by antibody testing; positive subjects were followed up by antigen testing and echocardiography (and necropsy if death occurred). The prevalence in dogs was 29% by a modified Knott test and antigen testing compared with a prevalence of 4.7% in cats by an antibody test; six of these infections (2.8%) were confirmed by the follow-up evaluations. This field study demonstrated that the prevalence of heartworm infection in cats in this area is within the expected limits of 9-18% of the prevalence in dogs. Antibody testing likely underestimates the real prevalence of D. immitis infection in cats. These results also emphasize the importance of preventive treatment in cats.     

Survey of heartworm prevention practices among members of a national hunting dog club

Surveillance data indicate that failures have been reported for virtually all heartworm prevention product categories. Resistance of third and fourth stage larvae of Dirofilaria immitis to macrocyclic lactones, lack of compliance, other unknown factors, or a combination of these reasons may be the cause of failure. A survey of members of a national hunting dog club was conducted to identify practices used to prevent canine heartworm infections. Questionnaires were completed by 708 dog owners. Year-round administration of heartworm preventive medication was reported by 208 (88%) respondents residing north of the 37th parallel. Dosing was based on the estimated weight of the dog by 54 (7%) respondents, 389 (55%) did not record the date prophylaxis was administered, and 89 (13%) observed the dogs spit out pills. Heartworm testing at least once per year was done by 556 (79%) respondents and test dates were spread throughout the year. Only 448 (64%) respondents tested newly acquired dogs for heartworm. These findings suggest that veterinarians should place a greater emphasis on the frequency and timing of heartworm diagnostic tests, the importance of weighing dogs, duration of administration, recording the date monthly heartworm prophylaxis is given, and observing dogs to ensure that oral medication is retained.     

Are antigen test kits efficient for detecting heartworm-infected dogs at the southern distribution limit of the parasite in South America? Preliminary results

The aim of this study was to evaluate the performance of commercial heartworm antigen tests in dogs harbouring Dirofilaria immitis microfilariae near its distribution limit in South America. A total of 4934 blood samples of adult dogs from Southern Greater Buenos Aires were examined to detect circulating microfilariae in the buffy coat interface between December 2005 and April 2006. Microfilariae were detected in 88 (1.8%) blood samples and all the microfilariae observed were identified as D. immitis by acid phosphatase stain technique. In a first trial, 69 (78.4%) out of the 88 serum were positive by Speed((R)) Diro. Then, a new test was performed over 25 microfilariae-positive serum samples randomly selected among the 88 previously tested samples and using simultaneously Speed((R)) Diro, Witness((R)) Dirofilaria and Snap((R)) 3dx. This second trial showed identical results for the three different tests, in which 19 (76%) samples were positive. Therefore, more than 20% of microfilaremic dogs were antigen negative. The main hypothesis that could explain our finding is a low worm burden in the study area. According to our preliminary results, it is highly recommendable the complementary use of antigen tests and other procedures to obtain an accurate diagnostic near the distribution limit of the parasite.     

Use of echocardiography for the diagnosis of heartworm disease in cats: 43 cases (1985-1997)

OBJECTIVE: To determine the usefulness of echocardiography in the diagnosis of heartworm disease in cats and to compare this modality with other tests. DESIGN: Retrospective study. ANIMALS: 43 cats with heartworm infection that had echocardiographic examinations at 2 veterinary teaching hospitals between 1985 and 1997. Twenty-two of these 43 cats also underwent radiography of the thorax and heartworm antibody and heartworm antigen testing. PROCEDURE: Cats were determined to be infected with Dirofilaria immitis infection on the basis of 1 or more of the following findings: positive modified Knott or antigen test result, echocardiographic evidence of heartworm disease, or confirmation of the disease on postmortem examination. The percentage of echocardiographs in which heartworms were evident was compared with the percentage of radiographs in which pulmonary artery enlargement was evident and results of antigen or antibody tests in cats in which all tests were performed. RESULTS: Overall, heartworms were detectable by use of echocardiography in 17 of 43 cats, most often in the pulmonary arteries. In the 22 cats in which all tests were performed, antibody test results were positive in 18, antigen test results were positive in 12, and pulmonary artery enlargement was evident radiographically and heartworms were identifiable echocardiographically in 14. Heartworm infection was diagnosed exclusively by use of echocardiography in 5 cats in which the antigen test result was negative. CONCLUSIONS AND CLINICAL RELEVANCE: Although echocardiography was less sensitive than antigen testing, it was a useful adjunctive test in cats that had negative antigen test results in which there was a suspicion of heartworm disease. The pulmonary arteries should be evaluated carefully to increase the likelihood of detection of heartworms echocardiographically.     

Epidemiology of canine heartworm in its southern distribution limit in South America: Risk factors, inter-annual trend and spatial patterns

This study was aimed at understanding some aspects of the canine heartworm epidemiology in the southern distribution limit of the parasite in South America. With this objective, 19,298 blood samples of owned dogs from 65 localities of 13 municipalities of Buenos Aires Province were tested for Dirofilaria immitis circulating microfilariae and/or female antigens. The overall heartworm prevalence was 1.63% by microhematocrit tube technique (n=19,136), 3.65% by modified Knott (n=713), and 14.41% by antigen test kit (n=118). Microfilaremic dogs showed a median of 1933 microfilariae per millilitre (q1=375, q3=5625, n=100). Male dogs belonging to breeds of short hair and large size recorded significantly higher prevalences than the other categories. Also, the prevalence increased significantly with the age and only dogs younger than 12 months were not found infected. A clear decreasing trend of the annual prevalence was observed during the whole study period, from 3.91% in 2001 to 1.17% in 2006. D. immitis-infected dogs were detected in 32 localities of 9 municipalities (prevalence range: 0.2-6.7%). Generalized linear models were used to assess associations between heartworm prevalence and environmental variables. The resulting significant models were univariate and included variables related with soil cover and human population density. The best model predicted maximum heartworm prevalences around middle values of bare soil cover, and lower at high and low covers. According to our analyses, canine heartworm infection in urban temperate Argentina could be described as relatively low, endemic, and spatially heterogeneous. Host and environmental factors affecting heartworm transmission at local level were identified and discussed.     

The safety-net story about macrocyclic lactone heartworm preventives: a review, an update, and recommendations

A number of safe, effective, and convenient heartworm preventatives are currently available for virtually all canine and feline pets. Yet, a 2001 survey of over 18,000 veterinary clinics in the United States identified more than 240,000 dogs and 3000 cats infected with Dirofilaria immitis. This high level of owner compliance failure is alarming. Prolonged administration of some of the macrocyclic lactone (ML) preventatives kills young larvae, older larvae, "immatures," young adults, and/or old adults. Efficacy of 95% or more requires dosing for 9-30 months, with older worms being more difficult to kill. Of the various MLs, ivermectin (IVM) has the most potent safety-net and adulticidal activity, milbemycin oxime has the least, and selamectin and moxidectin injectable lie somewhere in between. The unique effects of IVM are related to the age of the heartworms at initiation of treatment. The earlier treatment is started, the more stunted and smaller the worms and the shorter their survival time. Conversely, the later treatment is started, the longer the worms live, and the more likely the dog will be antigen- and microfilariae-positive. Drug effects do not appear to be enhanced by increasing the dosage or administering at shorter intervals, and it appears that continuous monthly treatment is needed to produce the full effects of the drug. The American Heartworm Society (AHS) recognizes the safety-net (or reach-back effect) and adulticidal properties of some MLs, particularly IVM. The AHS 2003 (American Heartworm Society, 2004. 2003 Updated guidelines for the diagnosis, prevention, and management of heartworm (Dirofilaria immitis) infection in dogs. In: McCall, et al., (Eds.), Proceedings of the Symposium Session on Recent Advances in Heartworm Disease, The 19th International Conference of the World Association for the Advancement of Veterinary Parasitology, New Orleans, LA, 10-14 August, 2003. Vet. Parasitol. 125, 105-130) canine guidelines state that it is beneficial to administer prophylactic doses of IVM before treatment with melarsomine. Results of laboratory studies suggest that less active dogs are at low risk of severe thromboembolism and death. However, heartworm-positive working dogs might be more at risk. Worsened radiographic and echocardiographic images in a client-owned dog given IVM monthly for 2 years with greatly restricted exercise suggests that such treatment of dogs with clinical, radiographic, and/or echocardiographic evidence of heartworm disease as well as for asymptomatic working dogs is contraindicated. Furthermore, until further data are available, such treatment of even the less active asymptomatic dog should be administered only with much caution and with examination by a veterinarian at least once every 4-6 months. IVM clearly provides potent "safety-net" activity against older larvae, immatures, and young adults in cases of owner compliance failure, even when the owner and veterinarian are not aware that the animal is infected, and offers much promise as a unique "soft-kill" treatment for young, and possibly older adult heartworms, with reduced risks.