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1.
The acid phosphatase activity and some morphological characteristics of Dipetalonema dracunculoides microfilariae are described. Their morphological features are closely related to those of the pathogenic Dirofilaria immitis when Knott's technique is used for the microfilarial diagnosis. The acid phosphatase activity pattern found in Dip. dracunculoides microfilariae is clearly different from those previously described for D. immitis, D. repens and Dip. reconditum.  相似文献   

2.
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.  相似文献   

3.
Using antigen capture and filter tests, 6,078 dogs throughout the state of Washington were examined for filariasis between July 1, 1997 and October 31, 1999. In western Washington, 791 males and 901 females examined were outdoors, not on prophylaxis, and had traveled out of the state; 6/791 (0.8%) males and 7/901 (0.8%) females were infected with Dirofilaria immitis (D. immitis), and one (0.1%) male and one (0.1%) female were infected with Dipetalonema reconditum (D. reconditum). There were also 392 males and 362 females examined that were outdoors, not on prophylaxis, and had not traveled out of western Washington. One (0.1%) female was infected with D. immitis, and two (0.5%) males and one (0.2%) female were infected with D. reconditum. In eastern Washington, 707 males and 826 females examined were outdoors, not on prophylaxis, and had traveled out of the state; 9/707 (1.0%) males and 4/826 (0.5%) females were infected with D. immitis, and no D. reconditum was found. There were also 376 males and 412 females examined that were outdoors, not on prophylaxis, and had not traveled out of the state. Three (0.8%) males and three (0.7%) females had D. immitis. One (0.2%) female had D. reconditum. Distribution of D. immitis-infected, nontravel dogs in eastern Washington was only found between 120 degrees and 119 degrees west longitude in the communities of Richland, Moses Lake, Okanogan, and Omak. Enzootic transmission of D. immitis and D. reconditum is occurring in both eastern and western Washington.  相似文献   

4.
Clinical records of 6,977 dogs examined at the small animal clinic of the University of Tennessee College of Veterinary Medicine from January 1980 through December 1989 were analyzed to determine the prevalence and changing frequency of Dirofilaria immitis and Dipetalonema reconditum infection. Using the Knott's test on these dogs, 805 (11.54%) tested positive for microfilariae, with D immitis in 430 dogs (6.16%), and Dip reconditum in 375 dogs (5.37%). Statistical analysis confirmed that the prevalence of D immitis and that of Dip reconditum were essentially equal in the population of dogs included in this study. There was a slight decrease in the prevalence of D immitis over the 10 years examined, but the prevalence of Dip reconditum remained constant. The results were not affected by year-to-year variability in the number of examinations performed. On the basis of our findings, in eastern Tennessee, those veterinarians who diagnose heartworm infection by the presence of microfilariae without differentiating the species involved, risk misdiagnosing 50% of the cases. If the patterns of prevalence seen in recent years continue, the chances of error may actually increase.  相似文献   

5.
A dog microfilariae prevalence and risk factor survey was conducted in 51 contiguous municipalities of the Mt. Vesuvius area (Campania region, southern Italy) in order to add data to the limited epidemiological information available regarding filarial worms in this zone. Between May 1999 and June 2000, blood samples were collected from 351 asymptomatic dogs. Blood samples were examined using a modified Knott's technique and histochemical staining in order to count and identify microfilariae. The results were subjected to statistical analysis and choroplethic municipal maps (MMs) were drawn by a geographical information system (GIS) software. Microfilariae were detected in 63 of the 351 dogs surveyed, constituting a total filarial prevalence of 17.9%. In particular, 56 dogs (15.9%) showed only microfilariae of Dipetalonema reconditum; three dogs (0.8%) only microfilariae of Dirofilaria repens; two dogs (0.6%) microfilariae of both D. reconditum and D. repens and two dogs (0.6%) microfilariae of both Dirofilaria immitis and D. repens. High D. reconditum prevalence was associated with hunting practice, masculine gender and older dogs. There was also a tendency to find high prevalence in dogs sampled in the afternoon.In conclusion, the presence of microfilariae of D. reconditum in 92% of microfilaraemic dogs indicates that this filarial worm was the predominant filarial species in dogs in the Mt. Vesuvius area.In addition, the general trends of the MMs showed that D. immitis and D. repens were present only in a few municipalities, whereas D. reconditum was widely and homogeneously spread throughout the entire study area.  相似文献   

6.
Both Dirofilaria immiti and Dipetalonema reconditum may be found in blood of infected dogs but it is not easy to distinguish D. immitis from D. reconditum in morphology. We cloned and sequenced the contiguous internal transcribed spacer (ITS) region, ITS1-5.8S-ITS2, of these two different parasites and published on GenBank as AF217800 for D. immiti and AF217801 for D. reconditum in this study. We designed two pairs of specific primers derived from ITS2 being used for polymerase chain reaction (PCR). The amplicons of ITS2 from D. immiti and D. reconditum are 302 and 348bp, respectively. Moreover, the limitation for amplifying ITS2 gene using this PCR demonstrated that 1 x 10(-2) microfilaria of each species of parasite smashed or even with mixed samples could be detected and the PCR products were predicted as the same as that described above. Thus, D. immiti and D. reconditum could be differentially diagnosed by this specific PCR. Seventeen clinical cases were evaluated and all of them were correctly identified. In this study, ITS1-5.8S-ITS2 of D. immiti or D. reconditum were the first time sequenced and analyzed. No significant similarity of ITS1 and ITS2 between D. immiti and D. reconditum could be observed.  相似文献   

7.
Screening of 25,822 dog blood samples indicated approximately a 1% infection rate with Dirofilaria immitis. Dipetalonema reconditum microfilariae were found in approximately 2% of 1,876 feral dogs examined. Laboratory experimentation indicated that 6 of 10 local mosquito species examined allowed successful extrinsic incubation of D immitis. Indices of experimental infection indicated Aedes triseriatus and Anopheles quadrimaculatus were excellent hosts for D immitis, but other factors considered, Aedes vexans probably served as the primary vector of D immitis. Coquillettidia perturbans and Culex pipiens-restuans group were considered unsuitable hosts and thus unlikely vectors of canine heartworm. Natural filarial infections were found in An quadrimaculatus and Ae vexans.  相似文献   

8.
Accurate diagnosis of canine filariosis is essential for choosing correct therapeutic approach. Therefore, reliable methods for discriminating among the different filarial infections in dogs are needed. The authors report simple and highly specific molecular methods that identify the three most common filarial nematodes of European dogs: Dirofilaria immitis, D. repens and Acanthocheilonema (syn. Dipetalonema) reconditum, based on (1) PCR amplifications of mitochondrial DNA (12S rDNA and coxI) with general filarial primers followed by digestion with restriction enzymes that generates band polymorphisms clearly discriminating the three species and (2) PCR amplifications with species-specific primers to support the restriction analysis, in particular in the case of multiple infections.  相似文献   

9.
Aedes albopictus and Culex quinquefasciatus were fed canine blood with different microfilarial density of Dirofilaria immitis ranging from 2500 to 25,000 mff/ml. Larval development in these two mosquito species did not differ significantly. Although C. quinquefasciatus ingested more microfilariae, the number of larvae which developed in A. albopictus was invariably greater than in C. quinquefasciatus. Mortality of the engorged A. albopictus was significantly greater than that of C. quinquefasciatus, and higher microfilarial density raised the mortality in both species. The vector efficiency index of A. albopictus was greater than C. quinquefasciatus at all microfilarial densities, but its survival time was much reduced. Thus, dogs with low microfilarial density are implicated as the main source for the transmission of D. immitis from dogs to mosquitoes.  相似文献   

10.
In 30 random-source dogs, we determined sensitivity and specificity of 5 serologic tests for detection of canine heartworm antigens. Seventeen of the dogs were infected naturally with adult Dirofilaria immitis, and 4 of the infected dogs were amicrofilaremic. The ability of the serologic tests to predict whether a dog was infected or uninfected (overall test accuracy) ranged from 73 to 97%. Sensitivity was not affected by circulating D immitis microfilariae, but was markedly influenced by the number of adult D immitis present. False-positive reactions were rare and were not associated with intestinal parasites or Dipetalonema reconditum microfilariae. Modifications of some of the test procedures were necessary to maximize test accuracy and reproducibility. These modifications and other technical details might limit the usefulness of some of the tests in a veterinary practice.  相似文献   

11.
Heart, lung and samples of blood from 230 dogs were examined for infections of filarial parasites. Dirofilaria immitis worms and microfilariae were detected in one dog. Blood samples from a further 1428 dogs were examined for microfilariae and 22 were found to be infected. Eighteen dogs were infected with D immitis microfilariae and four with Dipetolonema reconditum microfilariae. The histories were available for 18 of the dogs infected with heartworm and only seven dogs had not travelled outside South Australia. It was concluded that heartworm infection was endemic in South Australia but the apparent prevalence was only about 1%.  相似文献   

12.
This study describes a duplex real-time polymerase chain reaction (PCR) assay for the detection and differentiation between Dirofilaria immitis and Dirofilaria repens in dog blood and mosquitoes. Regions of a cytochrome oxidase 1 (cox1) mitochondrial DNA fragment and the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA were amplified from microfilariae and adult worm samples, using a sensitive SsoFast? EvaGreen(?) based real-time PCR method coupled with melting-curve analysis. The limit of the real-time PCR in detecting microfilaria and adult worm DNA was also tested both in dog blood and in artificially infected microfilarial. Two peaks at different melting temperatures (T(m)) for D. immitis (mean ± SD=75.7 ± 0.3°C) and D. repens (mean ± SD=70 ± 0.7°C), respectively, were obtained for microfilarial and adult positive controls of both species when examined separately and together. The real-time PCR protocol was also efficient in detecting microfilarial and adult DNA of both species when tested in samples spiked with DNA from Aedes albopictus, in Aedes aegypti experimentally infected by D. repens and in Culex pipiens naturally infected by D. repens and D. immitis. The high sensitivity of real-time PCR confirmed its reliability in detecting small amounts of genomic DNA either in dog blood or mosquitoes (2.5 pg/μl and 3 × 10(-1)pg/μl for D. immitis and D. repens, respectively). This assay is proposed as a tool for the epidemiological surveillance of the two most important Dirofilaria species in areas where they are endemic and sympatric.  相似文献   

13.
Microfilarial periodicity of Dirofilaria immitis in the venous blood of infected cats was analyzed by a trigonometric model. Cats were infected by subcutaneous transplantation with 120-day-old juvenile D. immitis. Microfilariae in the blood were first observed 98 days after transplantation. Blood was collected at 4h intervals for a 24h period, and examinations were repeated five times in two cats. The calculated periodicity index was 75.1 and 50.3 in these two cats. The estimated hour of peak microfilarial density ranged from 1.00 to 2.84h. Thus, the periodicity of microfilariae of D. immitis in the blood of cats was characterized as nocturnally sub-periodic.  相似文献   

14.
Blood samples from 331 greyhounds in the Hunter Valley and nearby coastal areas of New South Wales were examined for microfilariae using a filtration technique. Species were identified by histochemical staining; 10.9% of the greyhounds were infected with Dirofilaria immitis and 3.6% with Dipetalonema reconditum. The prevalence of infection of both species was significantly greater in summer than in winter (p = less than 0.05). Infection with D. immitis was correlated with differences in age, sex, bodyweight and coat colour, and a reported lack of stamina and the presence of a cough. No significant association was found. Diethylcarbamazine citrate was used for prophylaxis in 8.8% of all the greyhounds examined.  相似文献   

15.
The development of criteria for interpreting and reporting the results of an occult heartworm enzyme-linked immunosorbent assay to practitioners is described. The antigen is a saline extract of adult female Dirofilaria immitis. The cutoff absorbance A400 nm values were estimated, using 106 dogs free of infection. Any A400 nm value less than 0.526 is considered negative and values greater than or equal to 0.784 are positive. Intermediate A400 nm values are interpreted as suspect. Absorbance values for serum samples from 13 client-owned amicrofilaremic dogs revealed a bimodal distribution consistent with presumptive diagnosis based on clinical signs, which indicates that the test may be used to support a diagnosis of occult heartworm disease. The present serotest, however, is unable to distinguish microfilaremic dogs from noninfected dogs. Serum from dogs infected with other common helminths failed to crossreact with the D immitis antigen, with the exception of Dipetalonema reconditum.  相似文献   

16.
Canine dirofilariasis caused by Dirofilaria immitis is usually diagnosed by specific antigen testing and/or identification of microfilariae. However, D. immitis and at least six other filariae can produce canine microfilaremias with negative heartworm antigen tests. Discriminating these can be of clinical importance. To resolve discordant diagnoses by two diagnostic laboratories in an antigen-negative, microfilaremic dog recently imported into the US from Europe we developed a simple molecular method of identifying different microfilariae, and subsequently validated our method against six different filariae known to infect dogs by amplifying ribosomal DNA spacer sequences by polymerase chain reaction using common and species-specific primers, and sequencing the products to confirm the genotype of the filariae. We identified the filaria in this dog as D. repens. This is the first case of D. repens infection in the United States. Additionally, we examined microfilariae from five additional antigen-negative, microfilaremic dogs and successfully identified the infecting parasite in each case. Our diagnoses differed from the initial morphological diagnosis in three of these cases, demonstrating the inaccuracy of morphological diagnosis. In each case, microfilariae identified morphologically as A. reconditum were identified as D. immitis by molecular methods. Finally, we demonstrated that our PCR method should amplify DNA from at least two additional filariae (Onchocerca and Mansonella), suggesting that this method may be suitable for genotyping all members of the family Onchocercidae.  相似文献   

17.
OBJECTIVE: To determine prevalence of Dirofilaria immitis infection among shelter cats. DESIGN: Cross-sectional study. ANIMALS: 239 cats euthanatized at an animal shelter in southeastern Michigan. PROCEDURE: A gross necropsy focusing on the thoracic cavity, heart, lungs, and pulmonary vessels was performed within 5 hours after cats were euthanatized. Blood was collected directly from the heart of cats found to be infected and tested, using a filter test for microfilariae. Serum was tested for D immitis antigens by use of 2 antigen detection kits and for D immitis-specific antibodies by use of 2 antibody detection kits. RESULTS: Cats ranged from approximately 4 months to 15 years old. Adult D immitis were found in 6 (2.5%) cats. Blood could not be recovered from 1 of the cats with heartworm infection. For the 5 other cats, results of the filter test were negative, and results of both antigen and both antibody tests were positive. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cats living in an urban area in the northern part of the United States have a low prevalence of adult D immitis infection. However, this is likely to be an underestimate of the true prevalence of infection, because no attempts were made to identify cats infected with larval or juvenile stages of D immitis.  相似文献   

18.
A total of 174 dogs from an area hyperenzootic for Dirofilaria immitis were grouped into 4 age categories and necropsied; information was obtained on adult D immitis infections and on the presence of microfilariae. Serum samples from these dogs were examined by an enzyme-linked immunosorbent assay (ELISA) for antibody to adult D immitis and by an indirect fluorescent antibody test (IFAT) for antibody to microfilarial surface antigens. In dogs less than or equal to 5 months of age, necropsy demonstrated no evidence of infection; however, positive serologic results indicated that some of these dogs had prepatent infections. The percentage of dogs with ELISA titers (positive) increased with age, as did the percentage of dogs with adult D immitis infections. The IFAT results were positive in some dogs in each age category. Sera from all 29 dogs with occult infections were positive by ELISA. Sera from 6 of 20 dogs with occult dual-sex heartworm infections and 1 of 9 dogs with occult single-sex heartworm infections were positive by IFAT. For diagnosing occult dirofilariasis, the ELISA had a positive predictive value which increased with age of the dog to a maximum of 65.0% in dogs greater than or equal to 12 months of age; ELISA had a negative predictive value of 100% in all age groups. In contrast, positive and negative predictive values for the IFAT decreased with age of the dog to 60% and 37.5%, respectively, in dogs greater than or equal to 12 months of age.  相似文献   

19.
Dirofilaria immitis was diagnosed in 4 of 13 indigenous dogs from the Province of Zambézia, Mozambique, by acid phosphatase staining of microfilariae. The finding reconfirms the occurrence of the parasite in Mozambique after 3 decades and emphasises the need for extensive surveys. Additionally, in 1 of the infected dogs, microfilariae of Dipetalonema reconditum were detected, which is the 1st record of this parasite in Mozambique.  相似文献   

20.
A pruritic dermatitis characterized by the presence of erythema, papules, focal or multifocal alopecia, crusting and nodules was seen in 28 dogs with Dirofilaria repens microfilariae infestation in an endemic area in north-west Italy. Previous unsuccessful nonspecific antipruritic treatments, flea control and restricted diet were recorded in 53.6% of the patients. Both the Knott and the antigen tests were negative for Dirofilaria immitis and Acanthocheilonema reconditum. Concurrent babesiosis and/or canine granulocytic ehrlichiosis was also diagnosed in many affected dogs. Preliminary treatment of the concurrent diseases was followed by specific filaricide treatment. The cutaneous lesions, although slightly improved with the initial treatment, resolved completely with macro- and microfilaricide treatment. Although D. repens may be an opportunistic pathogen, this parasite should not be considered as harmless as previously thought and its potential pathogenic role in causing cutaneous lesions in dogs should be considered. Furthermore, it has a zoonotic importance as human cases have been reported worldwide.  相似文献   

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