Microfilaremic Dogs Research Articles

Detection of Wolbachia in Dirofilaria infected dogs in Portugal

Wolbachia pipiens, an intracellular endosymbiont bacteria of filarial nematodes, has been implicated in the pathogenesis of filarial diseases, in particular in heavy Dirofilaria spp. infections. Antibiotic therapy (doxycycline) against Wolbachia has been proven to be suitable adjunct therapy, prior to adulticide treatment of canine dirofilariosis. Despite its importance, investigation on the Wolbachia/Dirofilaria complex in Portugal had not been undertaken so far. This study reports the first detection of Wolbachia in Dirofilaria spp. infected dogs in the context of an ongoing epidemiological survey in central-south regions in the country. Wolbachia DNA was detected by PCR in 52.6% (20/38) of canine blood samples positive for Dirofilaria immitis based on parasitological (Knott's and Acid Phosphatase) and serological (Witness®Dirofilaria) methods. No Wolbachia DNA could be detected in samples from dogs with occult infections (parasite negative but antigen positive).The lack of Wolbachia detection in some microfilaremic dogs was somewhat unexpected and needs to be elucidated in further studies, as the presence or absence of these bacteria in association with microfilaria is of importance for veterinarians in the management and control of canine dirofilariosis.

Molecular detection of co-infections with Anaplasma phagocytophilum and/or Babesia canis canis in Dirofilaria-positive dogs from Slovakia

Recently, several arthropod-borne infections have been introduced into previously non-endemic regions in Europe as the result of various global changes. At the same time, endemic regions are expanding and the risk of co-infections is rising, due to climate change that allows vectors to move and spread infectious diseases into new areas. The aim of the current study was to confirm simultaneous infections with Anaplasma phagocytophilum and/or Babesia canis canis in Dirofilaria-infected dogs from Slovakia, central Europe. Genomic DNA was isolated from 366 blood samples of microfilaraemic dogs without clinical signs of infection. Samples were further screened for the presence of canine tick-borne pathogens using PCR and sequencing. This survey revealed co-infection with four arthropod-borne pathogens, in particular, Dirofilaria repens, Dirofilaria immitis, A. phagocytophilum, and B. canis canis. While D. repens, responsible for canine subcutaneous dirofilariosis, is scattered through the whole territory of the country, D. immitis occurs only in endemic areas of southeastern and southwestern Slovakia in mixed infection with D. repens. Co-infection with A. phagocytophilum was reported in 3.27% of the dogs positive for D. repens; mixed infection with D. repens and B. canis canis was detected in 3.55% of the tested blood samples. Eastern Slovak Lowland represents a natural focus of B. canis canis and is a highly endemic area for canine dirofilariosis. The presence of triple infection with D. repens, A. phagocytophilum, and B. canis canis was detected in one dog originating from the eastern lowland region of Slovakia. This study highlights the importance of co-infected, clinically healthy dogs in the spreading of several different arthropod-borne pathogens and the necessity for detailed epidemiological surveys, especially in newly infested areas.

Risk of canine and human exposure to Dirofilaria immitis infected mosquitoes in endemic areas of Italy

BackgroundThe occurrence of infections by Dirofilaria immitis in canine and human populations depends on several factors linked to both the definitive and intermediate hosts. Little data are available on the risk of human and dog exposure to D. immitis in endemic areas. Data collected on dog- and human-bait traps in endemic areas of north-eastern Italy were used to estimate the likelihood of a receptive host coming into contact with an infected vector.MethodsFrom 1997 to 1999, mosquitoes were collected from three sampling sites of north-eastern Italy on D. immitis microfilaraemic dogs and on human baits. The bite/night/host rates were determined based on the number of feeding and probing mosquitoes on dogs and humans, respectively. The survival/mortality rates of different species of mosquitoes following the blood meal, and the rate of natural Dirofilaria infection in unfed specimens were estimated. The risk of exposure of dogs and humans to infected mosquito species was determined by combining the bite/host/night and the mosquito infection rates.ResultsA total of 1,165 mosquitoes were collected on human (n = 815) and dog (n = 350) baits with varying species composition (i.e., Culex pipiens, 87.3% and Ochlerotatus caspius, 11.6%). Overall, dogs were more attractive to Cx pipiens than humans (feeding rate 70.2% vs probing rate 25.9%). The highest bite/night/host rate was 84.0 for dogs and 26.5 for humans. Cx pipiens displayed a mortality rate of 76.3% within 13 days and Oc. caspius of 100% within two days following the infective blood meal. In addition, D. immitis DNA was detected in unfed Cx pipiens (infection rate of 0.26%-2.07%). The infection rate adjusted for mosquito mortality was 0.38%. Based on data collected, the contact between an infected mosquito and a host can occur as often as every four nights for D. immitis infected-mosquitoes in dogs and within two weeks for humans.ConclusionsCx pipiens was confirmed as the most efficient natural vector of D. immitis in the studied area. In endemic areas, the risk of transmission can be very high for dogs and relevant for humans. Despite the increased awareness of veterinarians and owners on canine dirofilarioses, dogs from rural areas still maintain the natural life cycle of Dirofilaria spp., therefore acting as a source of infection to humans through vector bites.

Canine Dirofilaria Infections in Two Uninvestigated Areas of Serbia: Epidemiological and Genetic Aspects

In 2009 canine filarial infections were investigated in two northern areas of Serbia (Pančevo and Veliko Gradište), applying morphometry, biochemical staining, and immunological kit to detect Dirofilaria immitis antigens, and two home-made ELISAs to detect antibodies to D. repens and D. immitis somatic/metabolic polyproteins. Moreover, molecular tools were applied to analyze the phylogenetic relationships of the isolates. The microfilariae detected in 21/122 dogs (17.2%) were identified as D. repens (n=21) and D. immitis (n=2). D. immitis antigens were found in another 13 animals with occult infection. All of the 15 heartworm-positive dogs also had antibodies to this parasite, which were detected in another 13 subjects, indicating an overall D. immitis seroprevalence rate of 22.9%. Serology for D. repens revealed evidence of antibodies in 42.6% of the dogs, but was negative for 4 microfilaremic dogs. As for the two different areas, the prevalence of microfilariae and/or D. immitis antigens, mainly due to D. repens microfilaremic animals, was not significantly higher in Veliko Gradište (33.3%) than in Pančevo (22%). However, serology showed a different epidemiological picture. Heartworm infection occurred more often in both areas, and antibodies to dirofilarial nematodes were detected in 72.9% of dogs living in Pančevo, a rate higher than in those living in Veliko Gradište (57.1%). No risk factors for infection were found, confirming the uselessness of prophylactic drugs against D. repens, and suggesting the presence in these areas of sunrise- or sunset-biting mosquitoes as important vectors. The results indicate the need for both appropriate entomological studies and further research on the intra-species variability shown by D. repens.

New insights into the ecology and biology of Acanthocheilonema reconditum (Grassi, 1889) causing canine subcutaneous filariosis

In spite of its wide distribution among dogs and the evidence of its implication as a zoonotic agent, scant information is available on the biology of Acanthocheilonema reconditum (Spirurida, Onchocercidae). In this study, blood samples from 152 Sicilian dogs were examined for A. reconditum microfilariae at the beginning of the study and 1 year later. The periodicity of microfilaraemia was investigated by bleeding 2 highly microfilaraemic dogs twice a day for 10 days and, later on, every 2 weeks for 1 year and a third animal every 3 h for 96 h. Fleas and ticks infesting dogs were collected and dissected for the detection of A. reconditum larvae. The prevalence of infestation was 11·2% (17/152) and 13·3% (16/120) at the beginning and at the end of the study, with a 1 year cumulative incidence of 5·9%. Although dogs bled twice a day showed a higher number of microfilariae in most of the morning samples, the absence of any circadian rhythm was suggested by data of the third experiment conducted by bleeding a dog every 3 h for 4 days. A. reconditum developing forms were detected in 5·1% (4/78) of dissected fleas, but not in any of the 272 ticks. The study provides new insights into the biology and ecology of this dog filarioid in its definitive and intermediate hosts.

Dirofilaria immitis infection in dogs: Cardiopulmonary biomarker levels

Cardiopulmonary biomarkers are biological parameters that can be objectively measured and quantified as indicators of pathogenic processes (heartworm disease) or as indicators of response to therapeutic intervention. To determine levels of cardiopulmonary biomarkers in canine dirofilariasis, measurements of cardiac troponin T, cardiac troponin I, myoglobin, and D-dimer concentrations were performed for dogs with and without evidence of adult heartworm infection. The results showed that levels of cardiac troponin T were undetectable in all dogs studied while levels of cardiac troponin I were higher in dogs infected with Dirofilaria immitis. In healthy dogs, levels of myoglobin and D-dimer were below detection limits of the instrument and were significantly higher in heartworm-infected dogs, notably in microfilaremic dogs. The results suggest the possibility of using troponin I and myoglobin as markers for cardiac damage and the D-dimer as a supportive tool for a diagnosis of pulmonary thromboembolism in dogs with cardiopulmonary dirofilariasis.

Preliminary findings on the efficacy of selamectin in the treatment of dogs naturally infected with Dirofilaria repens

Subcutaneous dirofilariosis caused by Dirofilaria repens is common in dogs and it is an emerging helminthozoonosis in Europe, Asia, Africa and also in Hungary. Macrocyclic lactones are used for preventing the infection; however, their activity against the microfilariae and mature stages of this species is questionable. Selamectin is widely used for the prophylaxis of heartworm (D. immitis) infection. The objective of the present study was to test the microfilaricidal efficacy of the topical formulation of selamectin in dogs naturally infected with D. repens . A total of 78 Beagle dogs were examined for the presence of circulating microfilariae by Knott's test. Twenty-three of the microfilaraemic dogs were divided into four groups and included in the trial. The dogs received monthly or biweekly selamectin treatment and were subjected to monthly blood testing for a period of 252 or 336 days. At the end of the study, 65% of the dogs were not microfilaraemic and the rest had low number of microfilariae in their blood. These results indicate that chronic spot-on selamectin treatment may be a useful tool also in the control of canine subcutaneous dirofilariosis.

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.

Effects of doxycycline on the endosymbiont Wolbachia in Dirofilaria immitis (Leidy, 1856)—Naturally infected dogs

Dirofilaria immitis carries intracellular endosymbiotic bacteria of the genus Wolbachia, known to be vital for the worms and sensitive to tetracycline antibiotics. With the purpose of studying the interaction between D. immitis and the endosymbiont Wolbachia sp., heartworm naturally infected microfilaremic or antigenemic dogs were treated with doxycycline (10 mg/kg/day of the drug in three cycles of 21 days each, with 6-month intervals). Blood samples were collected on days 0, 7 and 21 of each treatment as well as on day 111 after the beginning of each cycle. A final sample was collected on day 723 from the beginning of the first treatment. The samples were examined for the presence and number of microfilariae and the presence of antigen as well as the presences of D. immitis and Wolbachia sp. DNA using PCR (polymerase chain reaction). With this approach, an evaluation of the effect of doxycycline on antigenemia and on the presence of Wolbachia sp. DNA in dogs with heartworm infection was possible. Doxycycline treatment did not alter the detection of adult parasite antigens with the exception of two animals, though the number of animals carrying Wolbachia sp. DNA decreased, despite the presence of the microfilariae. The effect of the antibiotic therapy on the worms may have interfered with the transmission of heartworm disease because the population of microfilariae and the number of microfilaremic dogs were reduced and the microfilariae positive samples that were found did not test positive for Wolbachia sp. in many cases. These findings suggest that in areas were doxycycline is extensively used D. immitis transmission may be impaired by the reduction on the number of microfilariae and on the endosymbiotic bacteria in the larvae turning them incapable of completing development once they infected a new host.

Incrimination of the mosquito,Aedes taeniorhynchus, as the primary vector of heartworm,Dirofilaria immitis, in coastal Yucatan, Mexico

Mosquito collections were carried out on microfilaraemic dogs, positive for Dirofilaria sp., for 18 consecutive nights in the coastal town of Celestún, Yucatan, southeast Mexico, during the rainy season (August) of 2007. A total of 292 female mosquitoes representing 12 species of dipteran Culicidae were collected: Anopheles albimanus (Wiedemann); Anopheles crucians (Wiedemann); Anopheles pseudopunctipennis (Theobald); Culex coronator (Dyar & Knab); Culex interrogator (Dyar & Knab); Culex nigripalpus (Theobald); Culex quinquefasciatus (Say); Culex salinarius (Coquillett); Aedes aegypti (Linnaeus); Aedes scapularis (Rondani); Aedes sollicitans (Walker), and Aedes taeniorhynchus (Wiedemann). Aedes taeniorhynchus and Cx. quinquefasciatus were the species found most commonly feeding on the dogs. Filarial nematodes were observed by microscopy in nine of the mosquito species collected; however, third-instar larvae were only observed in Ae. taeniorhynchus and An. crucians. Of 76 Ae. taeniorhynchus specimens found positive for Dirofilaria sp. by dissection, 14 were confirmed to be positive for Dirofilaria immitis (Leidy) by polymerase chain reaction (PCR). The resulting infection rate for D. immitis confirmed by PCR (6.2%) is higher than any infection rate for Ae. taeniorhynchus previously reported from the Americas.

Improved efficacy of tetracycline by acaciasides on Dirofilaria immitis

The discovery of Wolbachia, a bacterial endosymbiont that occurs in the filarial parasite and its sensitivity to tetracycline, has fostered a new initiative in the development of suitable antifilarial drugs. The present study is an attempt to investigate whether adding acaciasides (saponins from Acacia auriculiformis) to the standard dose of tetracycline would further improve the efficacy of tetracycline treatment against Dirofilaria immitis microfilariae in vivo. Treatment of microfilaremic adult dogs (body weight range 8-12 kg) with tetracycline at 10 mg/kg/day for 40 days resulted in 72% and 83% reduction in mf count on days 15 and 30, respectively, and the maximum reduction in mf count (91%) was achieved on day 75 post-treatment. However, treatment with tetracycline (10 mg/kg/day for 40 days) followed by acaciasides (10 mg/kg/day for 7 days) resulted in almost 100% clearance of mf at a faster rate on day 45 post-treatment and ensured long-term (until 4 months post-treatment) protection against microfilaremia. Data from polymerase chain reaction analysis reveals that compared to untreated dogs, in treated dogs, there was marked reduction in Wolbachia specific wsp markers in fast depleting mf population. The present data indicate that prior tetracycline treatment enhances microfilaricidal activity of saponins. This effect may be additive or synergistic as the worms are weakened by Wolbachia depletion, and these weakened microfilariae are possibly killed by the saponins.

Dirofilaria immitis (Leidy, 1856) no entorno de um caso felino: um estudo sobre sua transmissão

Dirofilaria immitis preferably infects canids and can, in lower frequency, be found in cats (Felis catus Linnaeus, 1758). The parasite may be transmitted by various Culicidae species. Motivated by an autochtone feline heartworm case diagnosed in the region of Engenho Novo - RJ (S 22 degrees 90' e WO 43 degrees 27'), the following issues were surveyed: 1) the local Culicidae fauna and; 2) the ocurrence of microfilaremic dogs. The mosquito study was conducted from April 2003 through May 2004, by peri-domiciliary and intra-domiciliary captures. A total of 672 mosquitoes (female) of the following species were caught: Aedes aegypti (Linnaeus) (561/672-83.5%); Culex quinquefasciatus Say (96/672-14.3%); Aedes (Oc.) scapularis (Rondani) (12/672-1.8%) and Aedes albopictus (Skuse) (3/672-0.4%). Only A. aegypti and C. quinquefasciatus specimens were captured year-round. A total of 235 canine blood samples were collected during the Campanha de Vacinação "Rio Sem Raiva" in October 2003. No microfilaria could be found in any of the examined samples (Knott s and NAN techniques), even though only 3.4% of the dogs received chemoprophylaxis and the majority of them frequently traveled to endemic areas. The presence of vectors associated to a possible dog mobility, may have provided the right scenario for the feline D. immitis infection to happen in the studied area.

<I>Ochlerotatus taeniorhynchus</I>: A Probable Vector of <I>Dirofilaria immitis</I> in Coastal Areas of Yucatan, Mexico

Mosquito collections were done on microfilaremic dogs, positive for Dirofilaria immitis (Leidy), for 15 consecutive nights in Celestun, Yucatan, southeastern Mexico, during January 2007. In total, 275 mosquitoes (3 male and 272 female) of five species were collected: Ochlerotatus taeniorhynchus (Wiedemann), Ochlerotatus sollicitans (Walker), Culex quinquefasciatus Say, Culex interrogator (Dyar & Knab), and Aedes aegypti (L.). Oc. taeniorhynchus was the species most frequently collected, and it had the highest rate of feeding success and the highest attack rates. First (L1) and third (L3) instars of Dirofilaria were observed in all mosquito species collected except for Ae. aegypti: 23 of 223 Oc. taeniorhynchus, three of 40 Cx. quinquefasciatus, and one of six Oc. sollicitans and one of one Cx. interrogator were infected with Dirofilaria (10.3% of total examined mosquitoes). This is the first report of Dirofilaria (presumably D. immitis) isolation from wild-caught mosquitoes in Mexico. Results imply that D. immitis can develop from microfilaria to infective L3s in Oc. taeniorhynchus, Cx. quinquefasciatus, and Oc. sollicitans, but L3s (infectious) were only recovered in heads of Oc. taeniorhynchus. Thus, Oc. taeniorhynchus can be considered a potential vector of dirofilariasis in Celestun, because infective L3 Dirofilaria were found in the vicinity of the mouthparts, and this mosquito has a strong association with microfilaremic dogs in this study area.

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 ® 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 ® Diro, Witness ® Dirofilaria and Snap ® 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.

A retrospective study of the clinical hematology and the serum biochemistry tests made on canine dirofilariasis cases in an animal hospital population in Bangkok, Thailand

Heartworm disease, caused by the filarial nematode, Dirofilaria immitis, is a major, potentially life-threatening disease of dogs, with worldwide distribution and global significance. It is not only of veterinary importance but it also has zoonotic potential in many regions. It is considered as an endemic disease in Thailand, although clinical data about the disease is rarely reported. The objectives of this study were to characterize the clinical hematology and the biochemistry of canine dirofilariasis cases admitted to Chulalongkorn University, Small Animal Teaching Hospital in Bangkok, Thailand, from 2001–2003. All hematology and serum biochemistry parameter interpretations were based on reference values. A total of 1023 dogs were evaluated in this study. Dogs were divided into three groups based on their heartworm classification (microfilaremic, occult and negative). The major hematological findings in microfilaremic dogs were a mild to moderate anemia, mild to severe thrombocytopenia, marked leukocytosis, moderate to marked neutrophilia, eosinophilia and monocytosis. The most common serum biochemical abnormalities in microfilaremic dogs included increased alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase with a mean ± SD of 311 ± 299, 82 ± 76 and 50 ± 38 IU/L, respectively. It is likely that this parasitic infection led to the impairment of the hematological and biochemical status of the infected dogs. We believe that investigation of these laboratory based parameters, when associated with the clinical signs, is a very important approach to be considered in the routine clinical follow up, as well as being important for therapeutic evaluations.

Discrimination between six species of canine microfilariae by a single polymerase chain reaction

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.

Epidemiology of heartworm: What is happening in South America and Mexico?

Canine heartworm has been known to exist in the Americas for more than 150 years (1847), and the first South American report was published approximately 30 years later (1875). The first human case was reported in 1887 and the first feline case in 1921. Coincidently, these two reports were from Brazil, where most surveys of the disease during the past two decades have been conducted in dogs. Feline cases are seldom found in the South American literature, although feline heartworm disease has been reported in Venezuela and Brazil. The prevalence in Mexico seems to be stable over the past 10 years (7.3–7.5%). Only 5 of the 13 South American countries have reported on canine heartworm infection prevalence. In Argentina, the national prevalence ranges from 3.5 to 12.7%; however, there was a much higher prevalence (17.7 and 23.5%) reported for two sections of Buenos Aires. In Lima, Peru, the prevalence is 4.35%, and Colombia reports 3.8–4.8% of dogs are infected. No infections were detected in 1056 dogs sampled in Chile. In Brazil, the national prevalence has declined from 7.9% in 1988 to 2% in 2001. The downward trend observed for canine heartworm in Brazil suggests that multiple pressures may be affecting the nematode transmission, including a possible decrease in mosquito vector population due to better control programs, reduction of transmission due to effective chemoprophylaxis, reduction of microfilaremic dog populations with the off-label use of injectable ivermectin, or reduction of the reproductive capacity of worms from tetracyclines used to control ehrlichiosis. Survey of a large population of captured mosquitoes indicated the most frequent species captured were vectors of heartworms ( Aedes taeniorhynchus (73.9%), Aedes scapularis (20%) and Culex quinquefasciatus (2.5%), suggesting that the composition of the mosquito population is not a critical factor in the decrease in heartworm infections in dogs in South America.

RELATIVE ABUNDANCE OF TREE HOLE–BREEDING MOSQUITOES IN BOONE COUNTY, MISSOURI, USA, WITH EMPHASIS ON THE VECTOR POTENTIAL OF AEDES TRISERIATUS FOR CANINE HEARTWORM, DIROFILARIA IMMITIS (SPIRURIDA: FILARIIDAE)1

Aedes (Protomacleaya) triseriatus currently shares its habitat in the USA with the introduced species Aedes (Finlaya) japonicus and Aedes (Stegomyia) albopictus. In the late 1980s, before the introduction of these 2 species, Ae. triseriatus was the dominant tree hole- and artificial container-breeding mosquito in central Missouri. Aedes triseriatus represented 89% of the mosquito immatures collected from water-filled tree holes and artificial containers at 3 forested field sites in central Missouri, from May to October, 1986 to 1988. Laboratory-reared female Ae. triseriatus were able to support larval development of Dirofilaria immitis (canine heartworm) to the infective 3rd larval stage. A blood meal from a microfilaremic Collie-mix dog was sufficient to infect adult female mosquitoes, indicating that Ae. triseriatus is a possible vector of canine heartworm in central Missouri. Confirmation of the vector status of this species depends on the yet-to-be observed transmission of D. immitis by Ae. triseriatus in the field, possibly by experimental infection of dogs by wild-caught mosquitoes. Defining the role of this species in epizootic outbreaks could contribute toward accurate risk assessment as the abundance of Ae. triseriatus increases and decreases in response to the success of Ae. albopictus, Ae. japonicus, or other introduced container-breeding mosquitoes.

A prevalence survey and risk analysis of filariosis in dogs from the Mt. Vesuvius area of southern Italy

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.

Immunological analysis of agglutination in Dirofilaria immitis microfllariae.

The mechanism of agglutination phenomenon of Dirofilaria immitis microfilariae was analyzed. Circulating microfilariae were collected from a D. immitis-infected microfilaremic dog and cultured in the several kinds of sera from dogs and animals. The agglutination of D. immitis microfilariae is a specific phenomenon due to some immune complexes formed with the anti-microfilarial antibody, heat-instable factor(s) and excretory-secretory products of microfilariae. Only live microfilariae were agglutinated and the agglutinated microfilariae remained alive as long as 27 days in culture in vitro.