Ocular dirofilariosis by Dirofilaria immitis in a dog: first case report from Europe

A five‐year‐old, entire female mixed‐breed dog was presented with corneal oedema and episcleral hyperaemia in the left eye. The ophthalmological examination revealed the presence of a free‐swimming nematode in the anterior chamber. Circulating microfilariae were not observed by a modified Knott test nor were adult antigens detected in serum by a commercial ELISA. The parasite was surgically removed from the dog's eye, but its anterior end was damaged during the surgery. Based on the morphology of the posterior end, the nematode was preliminarily identified as a male Dirofilaria immitis. The species identification was confirmed by PCR amplification and sequencing of the mitochondrial coxI and 12S rDNA genes, using a DNA barcoding approach. Although other cases of ocular dirofilariosis by D. immitis have been previously recorded in Australia and the United States, the case reported herein is the first in a dog from Europ.     

Aedes albopictus is a natural vector of Dirofilaria immitis in Italy

Investigations were carried out in Padova town (Veneto region, NE Italy) to define the actual role of Aedes albopictus in the natural transmission of Dirofilaria nematodes, and to assess the risk that its presence might represent for veterinary and medical health. During summer 2000-2002 daytime captures of human-attracted mosquitoes were carried out in three areas of the town. The presence of filarial parasites in mosquitoes was evaluated by PCR, and sequencing confirmed species assessment. DNA extraction was performed separately on pools of the insect abdomen and thorax-head, to discriminate between Dirofilaria infected/infective specimens. A total of 2721 mosquitoes were caught and A. albopictus was the most abundant species (2534). Filarial DNA was found in 27.5% (19/69) of the abdomen pools formed with mosquitoes collected in summer 2000, and in 11.1% (16/144) and 4.9% (6/123) thorax-head pools coming from samplings 2001 and 2002, respectively. Filarial DNA was belonging to D. immitis and all studied areas harboured infective specimens. These results prove A. albopictus as natural vector of D. immitis in Italy. Moreover, they support the hypothesis that the presence of the mosquito could affect the transmission pattern of canine heartworm disease in urban environment and, considering the aggressive anthropophylic behaviour of the species (30-48 bites/h) proven in Padova town, could enhance the circulation of filarial nematodes from animals to humans.     

Dirofilaria immitis and its potential mosquito vectors in central New York State.

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.     

The humoral response in natural Dirofilaria immitis infections in dogs

Plasma samples from dogs with infections of Dirofilaria immitis were assessed using ELISA and Western blotting techniques. These results were then assessed in relation to age and sex of the host, and to the numbers of microfilariae and adult filariae. Dogs with microfilariae tended to have lower levels of infection. In infected dogs, mean ELISA titres increased directly with the degree of infection. Dogs that were either young or mildly infected, showed a preferential antibody reactivity to antigens in the high molecular weight regions of the immunoblots. With increasing age and/or the extent of infection, an antibody response to antigens in the low molecular weight regions was apparent.     

Prevalence of Dirofilaria immitis in a stray dog population in western New York.

In a survey in 1973 undertaken to determine the prevalence of Dirofilaria immitis in 100 dogs from a public pound in western New York, 2 dogs were found to be positive. Microfilariae of Dipetalonema sp were not present. Hearts and pulmonary blood vessels were dissected for adult worms, and blood samples were examined for microfilariae, using a modified Knott technique. Although adult heartworms and microfilariae have been recovered from wild foxes, coyotes, and wolves in New York, this is the first report of heartworm in domesticated dogs in the state.     

Trichinellosis in Argentina: an historical review

In Argentina, Trichinella infection in pigs is endemic. The first report of human trichinellosis in Argentina was from 1898 in Buenos Aires. The number of human cases increased from 908, between 1971 and 1981, to 6,919, between 1990 and 2002. In pigs slaughtered in official establishments, the prevalence of Trichinella infection was 0.46% in 1914 and 0.01--0.03% during the period 1990--2004. T. spiralis is typically found in the domestic cycle that includes pigs, humans and rodents. Trichinella spp. from a sylvatic cycle has also caused human outbreaks resulting from the consumption of meat from puma, armadillo and wild boar. European migration to Argentina (principally Spanish and Italian) during the first years of the 20th century brought the tradition of preparing and eating raw sausages. This increased the risk of human exposure to Trichinella. Detection in pigs was initially made at slaughter by compression of muscle tissue (trichinoscopy) and continued this way until 1996, when artificial digestion was adopted for use in preventing human trichinellosis in Argentina. The following report synopsizes the evolution of trichinellosis in Argentina over the past century.     

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.     

Dirofilaria immitis infection in foxes and coyotes in Indiana

Between October 1976 and January 1977, carcasses or hearts and lungs from 202 wild foxes and coyotes in Indiana were collected to determine the prevalence of Dirofilaria immitis infection. Three of 113 (2.7%) red foxes (Vulpes fulva), 3 of 81 (3.7%) gray foxes (Urocyon cinereoargenteus), and 1 of 8 (12.5%) coyotes (Canis latrans) were infected with D immitis. An average of 5 worms (range, 1 to 10) were recovered from foxes and 11 worms were recovered from the coyote. Worms from the coyote were of normal length and all female worms contained developing embryos and microfilariae. Worms from foxes were small, and none of the female worms contained developing embryos.     

Prevalence of Dirofilaria immitis and Dipetalonema reconditum in greyhounds

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.     

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.     

Element composition and biochemical components of Dirofilaria immitis

Element composition of whole canine heartworms (Dirofilaria immitis) was determined using inductively coupled argon plasma emission spectroscopy. The elements evaluated included Na, K, Ca, Mg, P, Al, B, Ba, Cu, Fe, Mn, Sr, Zn, As, and Pb. The only significant sex difference was calcium concentration which was higher in females. Amino acid analyses of body fluid revealed presence of nine essential and eight non-essential amino acids. Concentrations for male and female heartworms were not significantly different. The spectrum of amino acids was similar to that previously reported for canine plasma. Neurotransmitters or their metabolites detected in whole male and female heartworms using HPLC with electrochemical detector were epinephrine, dihydroxyphenylacetic acid, dopamine, and 5-hydroxyindoleacetic acid. Those not detected were norepinephrine, homovanillic acid, and 5-hydroxytryptamine.     

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 prevalence of Dirofilaria immitis in dogs in Sydney

Four hundred and four dogs from 9 pounds in Sydney were examined for circulating microfilariae and antigens of Dirofilaria immitis. One hundred of these were also examined post mortem for adult heartworms. The prevalence of infection in the 404 dogs as shown by serology was 11.4%, and 5.9% had circulating microfilariae of D immitis. Adult heartworms were present in 15 of 100 dogs. Dipetalonema reconditum microfilariae were present in 3.7% of dogs. Dirofilariosis is still a common and important parasite of dogs in the Sydney region and chemoprophylaxis is recommended.