Candidatus Dirofilaria Hongkongensis Infections in Humans During 2005 to 2020, in Kerala, India.

We tried to determine the epidemiology and species of human dirofilariasis observed at two tertiary care hospitals in Kerala. We searched the hospital database to identify cases of dirofilariosis from January 2005 to March 2020. Along with human isolates, one dog Dirofilaria isolate was also subjected to PCR and sequencing of pan filarial primers cytochrome oxidase subunits 1 and 12S rDNA. We documented 78 cases of human dirofilariosis. The orbit, eyelid, and conjunctiva were the most commonly affected sites. Molecular characterization identified one dog and five human isolates as Candidatus Dirofilaria Hongkongensis. A rare case of subconjunctival infestation by B. malayi was also documented. Human dirofilariosis is a public health problem in the state of Kerala in India, and it is mostly caused by Candidatus Dirofilaria Hongkongensis. We propose that all diroifilaria isolates are subjected to sequencing for identification.

Dirofilaria hongkongensis infection presenting as recurrent shoulder mass.

In 2012, a novel canine Dirofilaria species, D. hongkongensis was identified in Hong Kong that caused human diseases and subsequently reported in an Austrian traveller returning from the Indian subcontinent. Here we present a case of human infection by D. hongkongensis manifested as recurrent shoulder mass. Diagnosis was achieved by cox1 gene sequencing of the excised specimen. The case illustrated that parasitic infection represents an important differential diagnosis for musculoskeletal lesions.

The Complexity of Zoonotic Filariasis Episystem and Its Consequences: A Multidisciplinary View.

Vector-borne transmitted helminthic zoonosis affects the health and economy of both developing and developed countries. The concept of episystem includes the set of biological, environmental, and epidemiological elements of these diseases in defined geographic and temporal scales. Dirofilariasis caused by different species of the genus Dirofilaria is a disease affecting domestic and wild canines and felines and man, transmitted by different species of culicid mosquitoes. This complexity is increased because Dirofilaria species harbor intracellular symbiont Wolbachia bacteriae, which play a key role in the embryogenesis and development of dirofilariae and in the inflammatory pathology of the disease. In addition, the vector transmission makes the dirofilariasis susceptible to the influence of the climate and its variations. The present review addresses the analysis of dirofilariasis from the point of view of the episystem, analyzing the complex network of interactions established between biological components, climate, and factors related to human activity, as well as the different problems they pose. The progress of knowledge on human and animal dirofilariasis is largely due to the multidisciplinary approach. Nevertheless, different aspects of the disease need to continue being investigated and cooperation between countries and specialists involved should be intensified.

[(EXPERIENCE WITH GEOGRAPHIC INFORRMATION SYSTEMS USED IN EPIDEMIOLOGICAL STUDIES (AS AN EXAMPLE OF MALARIA AND DIROFILARIASIS)].

The effective application of GIS against parasitic diseases requires the patterns of spread of parasitic diseases in certain natural, climatic, and socioeconomic conditions of different regions where there are epidemically effective carriers. The epidemiological parameters defining a high risk of an epidemiological process after P.vivax importation into Russia were calculated using the developed HealthMapper controlled module with a database. Analysis of the average long-term air temperatures in the administrative territories of Russia over 78 years, namely July, the warmest month of the year when there are the largest numbers of all types of mosquito vectors and high levels of microfilaria in the peripheral blood of in- fected dogs (definitive hosts), has shown that the northern border of the maximum possible area of dirofilariasis in Russia is most fully described by the +14C July isotherm.

Molecular characterization of human Dirofilaria isolates from Kerala.

Background & objectives: Human dirofilariosis is a well-recognized zoonosis caused by several species of the genus Dirofilaria. The disease is prevalent among canines and human beings in Kerala. The objective of the present study was to confirm the human Dirofilaria isolates by molecular characterization. Methods: The worms or segments obtained from human sources were subjected to diagnostic polymerase chain reaction (PCR) targeting the cytochrome oxidase subunit 1 (COI) and 5S ribosomal RNA (rRNA) genes of Dirofilaria repens. The amplicons were sequenced and analyzed. Results: The filariid nematodes recovered from ocular as well as subcutaneous tissue of human origin were identified as D. repens based on PCR targeting COI as well as 5S rRNA genes. The phylogenetic analysis of the COI gene nucleotide sequence obtained in the present study showed that D. repens shared the closest evolutionary relationship with D. honkongensis. Interpretation & conclusions: Molecular identification of D. repens isolated from human source assumes significance from the point of zoonotic threat of this mosquito-borne nematode. Phylogenetic analysis revealed a close evolutionary relationship with Asian isolate of D. honkongensis. Timely detection and treatment of infection in dogs, together with mosquito control, should be an integral part of the control strategy of this disease.

Regional warming and emerging vector-borne zoonotic dirofilariosis in the Russian Federation, Ukraine, and other post-Soviet states from 1981 to 2011 and projection by 2030.

We analyze through a climatic model the influence of regional warming on the geographical spreading and potential risk of infection of human dirofilariosis in Russia, Ukraine, and other post-Soviet states from 1981 to 2011 and estimate the situation by 2030. The model correctly predicts the spatiotemporal location of 97.10% of 2154 clinical cases reported in the area during the studied period, identified by a retrospective review of the literature. There exists also a significant correlation between annual predicted Dirofilaria generations and calculated morbidity. The model states the progressive increase of 14.8% in the potential transmission area, up to latitude 64 °N, and 14.7% in population exposure. By 2030 an increase of 18.5% in transmission area and 10.8% in population exposure is expected. These findings strongly suggest the influence of global warming in both geographical spreading and increase in the number of Dirofilaria generations. The results should alert about the epidemiological behavior of dirofilariosis and other mosquito-borne diseases in these and other countries with similar climatic characteristics.

Evaluation of the therapeutic and preventive efficacy of 2.5 % moxidectin / 10 % imidacloprid (Advocate(®), Bayer animal health) in dogs naturally infected or at risk of natural infection by Dirofilaria repens.

The objective of this GCP-compliant clinical field study was to evaluate the efficacy of the combination of moxidectin (minimum dose of 2.5 mg/kg body weight) and imidacloprid (minimum dose of 10.0 mg/kg body weight) spot-on (Advocate(®)) as a preventive and therapeutic treatment of natural infection by Dirofilaria repens in dogs in the Czech Republic.There were two arms of the study, both negatively controlled. 34 animals were randomly allocated to two groups of the treatment arm; 90 negative animals were randomly allocated to the prevention arm groups. All enrolled dogs were observed physically and blood was sampled monthly for Dirofilaria repens microfilaria counts for 18 months by modified Knott test and PCR. 34 dogs were positive for microfilaria and enrolled in the treatment arm of this study (treated: 18, untreated: 16). The reduction of the log-transformed microfilaria counts was significantly higher in the treatment group on day 28 (p = 0.007), 56, 84 and 112 (p < 0.001). All animals treated were negative after a single treatment. In the untreated control group 93.75 % remained positive (p < 0.001). 87 dogs were negative for microfilaria prior to allocation to the "preventive" arm (treated: 49; untreated: 38; 3 excluded). One dog in the untreated control group became positive for Dirofliaria repens microfilaria, while none of the treated dogs became positive. Advocate(®) was effective in the treatment of dogs infected with microfilaria of Dirofilaria repens. Due to the low rate of natural infections the preventive efficacy could not be proven, but no dog treated became positive.

Canine and feline infections by cardiopulmonary nematodes in central and southern Italy.

Capillaria aerophila, Aelurostrongylus abstrusus, Angiostrongylus vasorum and Dirofilaria immitis are cardiopulmonary nematodes affecting dogs and cats and presently emerging in several countries. The results obtained in 2009 - 2010 during a study aiming to investigate the occurrence of these nematodes in regions from Central (Marche and Abruzzo regions--Sites A and B, respectively) and southern (Apulia--Site C) Italy are here reported. A total of 534 and 436 individual faecal samples collected from dogs and cats were examined, together with 471 and 34 faecal environmental samples taken from dog shelters and catteries. One hundred and ninety-two individual blood samples were also collected from dogs. Faeces were examined using copromicroscopical flotations and Baermann technique, whereas blood samples were tested by Knott's method. Eggs of C. aerophila were detected in 1.48 % and 20 % (Site A), 8.67 % and 2.71 % (Site B), and 16.67 % and 0 % (Site C) of individual and environmental canine samples. C. aerophila was found in 2.90 % (Site A), 3.03 % (Site B) and 14.29 % (Site C) of individual cats. Larvae of A. vasorum were found in 0.96 % and 2.48 % of individual and environmental samples from Site B, respectively, while those of A. abstrusus in 1.82 % (Site A) and 9.96 % (Site B) of individual faeces. Microfilariae of Dirofilaria immitis, identified on the basis of key morphological and morphometric features, were detected in 2.56 % of samples collected from Site B. Despite the small sample size, these results indicate that cardiopulmonary nematodes occur in Central and southern Italy, thus they should be included in the differential diagnosis of pet cardiorespiratory diseases. Larger surveys are necessary to gain more information on the diffusion of these parasites, especially for C. aerophila and A. vasorum, for which the actual distribution is poorly known.

Changing climate and changing vector-borne disease distribution: the example of Dirofilaria in Europe.

Climatic changes, together with an increase in the movement of dogs across Europe, have caused an increase in the geographical range of Dirofilaria infections. The present paper is focuses on northeastern European countries, where survey data have shown an increase of Dirofilaria repens infections both in animals and humans. A growing degree day-based forecast model has been developed to predict the occurrence. The model is based on evidence that there is a threshold of 14 °C below which Dirofilaria development will not proceed in mosquitoes, there is a requirement of 130 growing degree-days (GDDs) for larvae to reach infectivity, and there is a maximum life expectancy of 30 days for a mosquito vector. The output of this model predicted that the summer temperatures (with peaks in August) are sufficient to facilitate extrinsic incubation of Dirofilaria even at latitudes of 56 °N and longitudes of 39 °E. Despite the fact that both Dirofilaria immitis and D. repens have the same temperature requirement for extrinsic incubation in mosquitoes, empirical data has shown that D. repens is the main cause of dirofilarial infections in both humans and animals. Clinical signs are absent in most canine infections with D. repens. Furthermore, diagnosis is problematic and in-clinic serological tests, such as those for D. immitis, do not exist. Therefore, most infections go undiagnosed, allowing the infection to spread undetected.

What is new about animal and human dirofilariosis?

Dirofilaria immitis and D. repens, the causal agents of cardiopulmonary and subcutaneous dirofilariosis, respectively, affect canine, feline and human populations with an increasing incidence in temperate and tropical areas of the world. In the past decade outstanding advances in the knowledge of dirofilariosis have been achieved. Nevertheless, questions such as the impact of climate change in the transmission and distribution of dirofilariosis, as well as a profound evaluation of both the role of Dirofilaria and Wolbachia and the proteins produced by them in the parasite-host relationship have not been fully addressed; therefore there must be milestones in dirofilariosis research in order to design new strategies and tools for the control of this disease.

Wolbachia in Dirofilaria repens, an agent causing human subcutaneous dirofilariasis.

Human subcutaneous dirofilariasis is an increasingly reported zoonosis caused by several filarial species, in particular by Dirofilaria (Nochtiella) repens. Like many filarial worms, D. repens harbors the bacterial endosymbiont Wolbachia that has been implicated in the inflammatory features of filarial infection. Immunohistochemical staining against the Wolbachia surface protein (WSP) was carried out on 14 skin nodules and showed numerous bacteria within the intact worms and occasional positive staining within the surrounding inflammatory infiltrate. Serum samples from 11 of these patients resulted positive for total immunoglobulin G titers against WSP as examined in enzyme-linked immunosorbent assay. This is the first description of Wolbachia distribution in D. repens and the first report of specific immune response to Wolbachia in patients with subcutaneous dirofilariasis.

Allergic inflammatory reaction is involved in necrosis of human pulmonary dirofilariasis.

AIMS: To determine whether, in view of the massive inflammatory cell infiltration and the rounded rather than wedge-shaped character of pulmonary lesions in dirofilariasis, the inflammatory response against the worm contributes to the coagulative necrosis, in addition to an ischaemic process. METHODS AND RESULTS: The histopathological features of 13 resected dirofilariasis cases with well-defined nodules ranged from 10 to 30 mm were analysed. On routine histology and using immunohistochemistry, the peripheral encapsulating wall showed mild to severe infiltration of eosinophils, lymphocytes and plasma cells and a histiocytic reaction in all cases, often with necrotic eosinophils seen within the necrosis (84.6%) and inflammatory changes in the adjacent lung (38.5%). The CD4+ lymphocyte count (80.8 +/- 33.4) was greater than that of CD8+ lymphocytes (24.5 +/- 16.9) in the central necrosis and vice versa in the wall. In the necrotic regions, disruption of the pulmonary artery (61.5%) and extravasation of the torn worm (23.1%) could be seen. CONCLUSIONS: These findings indicate that an allergic inflammatory reaction, mediated by eosinophils and lymphocytes, is involved in the formation of the dirofilarial necrotizing granuloma rather than infarction caused simply by embolism.

Intravitreal dirofilariasis: a rare ocular infection.

Human ocular dirofilariasis is a zoonotic disease, rare in Europe, caused by filarial nematodes. The parasite is either encysted in a subcutaneous nodule or located under the bulbar conjunctiva. We report the case of a 62-year-old man with intravitreal dirofilariasis, which is a rare site of presentation of the nematode in the human eye. It was located in the fundus area and was surgically removed. The nematode was identified as Dirofilaria repens (D. conjuctiva) by two different Microbiology Departments, making this the fifth report of identified intravitreal dirofilariasis caused by D. repens in the relative literature.

Retrospective study on the presence and pathogenicity of Dirofilaria repens in 5 dogs and 1 cat from Aosta Vally.

Aosta Valley is the only region of Italy free of reported human cases of dirofilariosis due to Dirofilaria repens. Nonetheless, the retrospective analysis of five locally acquired D. repens infections in dogs, seems to indicate that an animal reservoir exist in the area. A feline case imported from Camargue, France, is also described. The most common dermatological signs were erythema, alopecia, crusting, papulae and nodules. All patients had lesional pruritus and dogs were affected by concurrent babesiosis. Eradication of the underlying condition, followed by a therapy with the specific adulticide and microfilaricide drugs led to a complete recovery of microfilaraemia. Increased pet travel and ecological changes are extending the ranges of vector borne, parasitic and zoonotic diseases. This fact pose a unique diagnostic challenge for the veterinarian, since the index of suspicion may be absent outside endemic areas.

Dirofilariasis of breast: report of two cases diagnosed by fine-needle aspiration biopsy.

We report on 2 cases of dirofilariasis of the breast occurring in adult Chinese females residing in Hong Kong. The diagnosis was made by fine-needle aspiration biopsies. The direct smears contained numerous acute inflammatory cells, including eosinophils. A partially necrotic adult nematode of about 400-450 microm in diameter was identified in the cell-block sections of each case. The worm was characterized by a thick cuticle with longitudinal cuticular ridges and fine transverse surface striations, abundant somatic muscle, an intestine, and a reproductive tube or uterus. The morphologic features of this round worm were most compatible with those of Dirofilaria repens. While the clinical and radiologic findings of dirofilariasis of the breast are nonspecific and mimic those of breast neoplasms or inflammatory conditions, a definitive diagnosis is possible with fine-needle aspiration biopsy. A correct diagnosis of dirofilariasis may help to spare patients from unnecessary medical and surgical interventions.

Dirofilaria repens infection in a dog in Israel.

Only one case of canine infection with Dirofilaria repens has been reported in Israel to date; this occurred in 1934. This publication, 65 years later, represents the second case of D. repens infection in a dog in Israel. This dog was infected locally since it was born in Israel and was never taken abroad, suggesting that the life cycle of the filaria was completed in Israel. Since dogs, cats, and foxes serve as a reservoir for the filaria and these are abundant in Israel, and mosquitoes of the genera Culex and Aedes (both of which occur in Israel and the Middle East) are vectors, the conditions for establishment of the filaria in Israel exist, and warrant regional epidemiologic investigation.

Adult Dirofilaria repens in a subcutaneous granuloma on the chest of a dog.

A 10-year-old, 9 kg, intact male crossbred dog was treated for nasal mites with milbemycin oxime using a dose of 1 mg/kg bodyweight orally, three times at 10-day intervals. One month after the initiation of this treatment a subcutaneous nodule developed on the sternum of the dog. The nodule was removed and found to contain a single, 82 mm long, thread-like nematode. Several exotic parasites were suspected as possibilities because the dog had been imported to Norway from South Africa. Microfilariae were not detected in the blood and heartworm antigen tests were negative. The worm was identified morphologically as an adult, female Dirofilaria repens. This is the first report of D repens from Norway. The case is of interest because of the differential diagnostic problem it posed and because infestation was recognised following treatment of another parasitic condition with a broad-spectrum, antiparasitic drug. In addition, the case provides a reminder of the necessity to be aware of geographical differences in disease occurrence which can produce unexpected disease in non-endemic areas as a consequence of increased international travel with pets.

Importance in France of the infestation by Dirofilaria (Nochtiella) repens in dogs.

The presence of Dirofilaria repens has been mentioned in Europe, in Africa and in Asia, in different carnivores (dogs, foxes, cats, lions). In Europe the nematode has been reported in dogs, cats and foxes and the countries concerned by native cases are Italy, Spain, Greece, Yugoslavia and France. In France, until now, D. repens has been described, among animals, only in dogs. The first undoubted report dates back to 1953. Later, the prevalence was studied through widespread surveys (1986-1988-1989), focal surveys within some canine populations (military dogs) and some sporadic and fortuitous descriptions. The prevalence has been mainly assessed by pointing up the microfilariae in the blood and by identifying it after histochemical staining (for demonstration of the somatic distribution of acid phosphatase activity). The infestation rate was higher than 20% in some military dog populations. The cases occurred in nineteen French departments (Alpes-de-Haute-Provence, Aude, Bouches-du-Rhône, Corrèze, Corse, Dordogne, Gard, Gers, Gironde, Haute-Garonne, Loir-et-Cher, Nièvre, Pyrénées-Orientales, Rhône, Tarnet-Garonne, Var, Vaucluse, Vienne and Region Parisienne). The pathogenicity which is normally denied has to be reconsidered. The pathology of adult worms has not been clearly proved in dogs but the embolized microfilariae can bring about skin troubles. The potential pathogenicity, the necessity of the differentiation with D. immitis and above all, the zoonotic importance of D. repens must lead to search for it more often and to consider it with great interest.