Leishmania infantum and Dirofilaria immitis infections in Italy, 2009-2019: changing distribution patterns.

BACKGROUND: For long time, canine leishmaniosis (CanL) was considered endemic in the southern, central, and insular regions of Italy, whereas heartworm disease (HW) caused by Dirofilaria immitis was considered endemic in the northern region and in the swampy Po Valley. Following the reports of new foci of both diseases, in this study we update the distribution patterns and occurrence of new foci of CanL and HW discussing the main drivers for the changes in the epidemiology of these two important zoonotic canine vector-borne diseases. METHODS: Based on the statistical analyses of serological assays (n = 90,633) on L. infantum exposure and D. immitis infection performed by two reference diagnostic centres in Italy over a ten-year period (2009-2019) irrespective of the anamnesis of dogs. The distribution patterns of both parasites are herein presented along with the occurrence of new foci. RESULTS: Results highlighted the changing distribution patterns of L. infantum vs D. immitis infection in Italy. CanL is endemic in some areas of northern regions and HW has endemic foci in central and southern regions and islands. Significant differences in L. infantum exposure and HW infection prevalence among the study macroareas were detected. The overall results of the positive tested samples were 28.2% in southern Italy and islands, 29.6% in central Italy and 21.6% in northern Italy for L. infantum and 2.83% in northern Italy, 7.75% in central Italy and 4.97% in southern Italy and islands for HW. HW positivity significantly varied over years (χ2 = 108.401, df = 10, P < 0.0001), gradually increasing from 0.77% in 2009 to 8.47% in 2016-2017. CONCLUSIONS: New potential epidemiological scenarios are discussed according to a range of factors (e.g. environmental modifications, occurrence of competent insect vectors, transportation of infected animals to non-endemic areas, chemoprophylaxis or vector preventative measures), which may affect the current distribution. Overall, the results advocate for epidemiological surveillance programmes, more focussed preventative and control measures even in areas where few or no cases of both diseases have been diagnosed.

Mosquito diversity and dog heartworm prevalence in suburban areas.

BACKGROUND: Urbanization is occurring rapidly on a global scale and is altering mosquito communities, creating assemblages that are characteristically less diverse. Despite high rates of urbanization and ample examples of vector-borne diseases transmitted by multiple species, the effects of urbanization-driven mosquito diversity losses on disease transmission has not been well explored. We investigated this question using the dog heartworm, a filarial parasite vectored by numerous mosquito species. METHODS: We trapped host-seeking mosquitoes in undeveloped areas and neighborhoods of different ages in Wake County, North Carolina, USA, analyzing captured mosquitoes for heartworm DNA. We compared within-mosquito heartworm infection across land-use types by Kruskal-Wallis and likelihood ratio tests. Using zip code level data acquired from dogs in a local shelter, we performed linear regressions of within-host heartworm prevalence by within-mosquito heartworm prevalence as well as by three mosquito diversity measures. We also determined the best predictor of host-level prevalence among models including within-mosquito infection, mosquito diversity and abundance, and socioeconomic status as variables. RESULTS: Suburban areas had lower within-mosquito heartworm prevalence and lower likelihood of heartworm-positive mosquitoes than did undeveloped field sites, although no differences were seen between suburban and undeveloped wooded sites. No relationships were noted between within-mosquito and within-host heartworm prevalence. However, mosquito diversity metrics were positively correlated with host heartworm prevalence. Model selection revealed within-host prevalence was best predicted by a positive relationship with mosquito Shannon-Wiener diversity and a negative relationship with household income. CONCLUSIONS: Our results demonstrate that decreases in mosquito diversity due to urbanization alter vector-borne disease risk. With regard to dog heartworm disease, this loss of mosquito diversity is associated with decreased heartworm prevalence within both the vector and the host. Although the response is likely different for diseases transmitted by one or few species, mosquito diversity losses leading to decreased transmission could be generalizable to other pathogens with multiple vectors. This study contributes to better understanding of the effects of urbanization and the role of vector diversity in multi-vectored pathosystems.

Diagnóstico de Dirofilaria immitis en el municipio Sucre, estado Sucre, Venezuela / Diagnosis of Dirofilaria immitis in the municipality of Sucre, Sucre state, Venezuela

La dirofilariasis es una patología parasitaria, causada por los nematodos Dirofilaria immitis y D. repens, principalmente. Mientras que D. immitis presenta distribución mundial, D. repens se extiende por el viejo mundo. Ambas especies afectan a caninos y felinos domésticos y salvajes, así como a humanos, por lo que es un problema de salud pública mundial. En este estudio se determinó la prevalencia de D. immitis, en caninos del municipio Sucre, estado Sucre, Venezuela. Se tomaron muestras sanguíneas a 138 caninos domésticos, mayores de seis meses, sin distinción de raza ni sexo, previo consentimiento informado por sus propietarios. El diagnóstico parasitológico de las microfilarias circulantes se realizó por examen directo y método de concentración Knott modificado; las características morfológicas fueron observadas en extendidos teñidos con Giemsa. Se utilizó un estuche comercial, para la captura de antígenos de excreción-secreción de D. immitis. De los 138 caninos estudiados, 12 casos positivos (8,7%) fueron detectados parasitológicamente, mientras que por el estuche comercial se detectaron antígenos en 20 caninos (14,5%), lo que demuestra que existe una dirofilariasis oculta en el 5,8% de los casos evaluados. Uno de los perros microfilarémicos, no presentó antígenos circulantes. La prevalencia de dirofilariasis en el municipio Sucre, estado Sucre fue de 15,2%. Este reporte de dirofilariasis canina en el municipio Sucre, representa un importante aporte en el levantamiento epidemiológico de la enfermedad. Futuros estudios permitirán evaluar el riesgo de infección de las personas que conviven con los perros con dirofilariasis.

Heartworm is a parasitic disease, caused by nematodes Dirofilaria immitis and D. repens, mainly. While D. immitis presents worldwide distribution, D. repens extends the old world. Both species affect canines and felines domestic and wild, as well as human, so it is a global public health problem. In this study we determined the prevalence of D. immitis in dogs in the municipality of Sucre, Sucre state, Venezuela. Blood samples were taken for 138 domestic dogs, over six months, regardless of race or sex, prior informed consent by their owners. The parasitological diagnosis of circulating microfilariae was done by direct examination and concentration method modified Knott, the morphological characteristics were observed in Giemsa-stained smears. We used a commercial kit, for the capture of excretory-secretory antigens of D. immitis. Of the 138 dogs studied, 12 cases (8.7%) were detected parasitologically, whereas the commercial kit antigens were detected in 20 dogs (14.5%), demonstrating that there is an occult infections in 5.8% of the cases. One of the dogs with microfilariae in blood, did not submit antigens circulating. The prevalence of heartworm in the municipality of Sucre, Sucre state was 15.2%. This report of canine heartworm in the municipality of Sucre, represents an important contribution in an epidemiologic survey of the disease. Future studies will assess the risk of infection for people living with dogs with heartworm disease.

Activity of pulmonary intravascular macrophages in cats and dogs with and without adult Dirofilaria immitis.

Pulmonary intravascular macrophages (PIMs), large (20-80 microm diameter) monocytes are present in sheep, pigs, and horses, but not in dogs, rats, rabbits, or primates. The present study evaluated the phagocytic activity of various organs in cats and dogs and determined the influence of Dirofilaria immitis infections on PIM activity. Live or dead adult heartworm (HW) was transplanted via jugular venotomy into cats and dogs. Cats (four per group) were allocated to five groups: surgical controls--no HW, dead HW for 1 week, live HW for 1 week, dead HW for 3 weeks, or live HW for 3 weeks. Radioactive technetium (Tc-99m, 1.2mCi in 0.3ml) sulfa-colloid was injected intravenously. All cats with HW were clinically asymptomatic and developed radiographic pulmonary parenchymal changes. No gross changes were visible at necropsy for cats with HW; inflammatory changes were less severe in cats with live HW. In cats with dead HW for 3 weeks, worms were present but folded, flattened, and located in distal pulmonary arteries. Uninfected control dogs and those with dead HW did not demonstrate any PIM activity. In control cats, lungs were the primary phagocytic organ after systemic IV colloid injection (72.5% of the total recovered radioactive dose). The lung and liver together represented over 95% of the recovered Tc-99m colloid in all cats. In each group of cats with HW, phagocytic activity of the lung was significantly less (p < 0.001) than the PIM activity of controls. Cats with dead HW at 1 week (50.1%) had a significant (p < 0.019) decrease in PIM activity compared with cats with dead HW at 3 weeks (59.5%). The PIM activity in cats with live HW was significantly decreased (p < 0.001) from that in groups with dead HW, but there was no significant difference between the two groups infected with live worms. There were no significant differences in recovery between any groups in pairwise analysis of the spleen, heart, skeletal muscle, kidney, bone marrow, or blood. Significant increases (p < 0.001) in liver activity for each group inversely reflected the decreased lung activity; consistent with increased hepatic uptake of Tc colloid "escaping" a relatively suppressed lung macrophage system. Transmission electron microscopy confirmed PIM glycocalyx changes and vacuolization, moderate Type 1 cell damage and Type II cell hypertrophy in cats with dead HW. There was no evidence of PIM death. The significant decrease in PIM activity in groups with dead HW and a greater decrease in groups with live HW are consistent with a down-regulation of PIM function in cats with live HW.

The epidemiology of Dirofilaria immitis infection in outpatient dogs at Chiang Mai University Small Animal Hospital, Thailand.

Five hundred eighty-nine dog blood samples from the small animal hospital of Chiang Mai University were examined for Dirofilaria immitis prevalence using a microhematocrit tube technique for microfilaria detection. In parallel, a once a month follow-up study on 36 D. immitis negative dogs was conducted to detect the time of acquiring infection in each animal. The diagnostic criteria for the incidence study was based on microfilaria detection or on positive findings against D. immitis antigen using the Witness commercial kit. The estimated prevalence was 18.2% (15-21%; 95% CI). There was no statistical difference between male and female infection rates. The age-specific prevalence of dogs under 2 years old was 6.4%, which was lower than the 2-4 year old group and all the other age groups at a 95% confidence level. In older dogs the prevalence reached 41.5%. Most of the dogs housed outdoors had a statistically higher infection rate than the dogs housed indoors (chi-square = 9.662, 1 df, p = 0.002). Only 109 dogs received chemoprophylaxis resulting in a significantly lower infection rate than in the non-heartworm prevention dogs (chi-square =14.424, 1 df, p = 0.000). The overall incidence density and the incidence during the rainy, cool and hot seasons were 5.2, 6.9, 3.5, and 2.7 animals per 100 animal-months, respectively. The incidence rate ratio between wet/dry, rainy/cool, rainy/summer, and cool/hot seasons were 2.18, 1.98, 2.59 and 1.30, respectively. The 95% confidence interval revealed no difference among seasons. In conclusion, dogs in D. immitis endemic northern Thailand contract infection in about 2 years.

IgG response against infective larvae of Dirofilaria immitis in experimentally infected cats.

Somatic antigens from third stage larvae of Dirofilaria immitis (SL3) were used to detect IgG response against heartworm infection in 8 experimentally infected cats. A moderate specific anti-SL3 IgG response was found one month post-infection. Afterwards, antibodies decreased reaching a basal level 4 months post-infection and remained at this level until the end of the study. 6 months post-infection. Western blot analysis showed specific recognition of polypeptides of 79, 73, 60, 52, 40 and 39 kDa by sera from infected cats 1 month post-infection, but not by sera taken prior to the infection. The low antigenicity of the SL3 antigen in the cat should allow the parasite to escape the host's immune response.

Potencial vectors of Dirofilaria immitis (Leidy, 1856) in Itacoatiara, oceanic region of Niterói municipality, State of Rio de Janeiro, Brazil

Dirofilaria immitis is a widespread mosquito-borne parasite that causes dirofilariasis, a commonly diagnosed disease of dogs that is rarely reported in cats and humans. A mosquito survey was conducted in Itacoatiara in the State of Rio de Janeiro, from March 1995 to February 1996, using canine, feline and human baits. A total of 3.667 mosquitos were dissected for D. immitis larvae, representing 19 species and 10 genera. From those, Ae. scapularis, Ae. taeniorhynchus, Cx quinquefasciatus, Cx. declator, Cx. saltanensis and Wy. bourrouli were found infected with D. immitis parasites, and among those, only the first three harbored infective larvae. The majority of larvae were found in the Malpighian tubules (889/936), and larval melanization was observed in the two Aedes species. In descending order, the best vector were Ae. scapularis, Ae. taeniorhynchus, and Cx. quinquefasciatus which alternate seasonally in importance. Cx. quinquefasciatus is suggested to be a vector to cats. The potential transmission of D. immitis parasites by these three vectors to man is discussed.

Purification and characterization of a neutrophil chemotactic factor from Dirofilaria immitis.

A neutrophil chemotactic factor (NCF-Di) was purified from a crude extract of Dirofilaria immitis adult worm by a combination of anion-exchange chromatography on DE52 and gel filtration on Sephacryl S-200. NCF-Di showed a single protein band by both polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate (SDS) PAGE. The molecular weight of NCF-Di was estimated to be 17,000 by gel filtration on Sephadex G-150, and 14,000 by SDS-PAGE. NCF-Di was an acidic protein with isoelectric point of 4.5. NCF-Di was absorbed neither to lentil lectin-Sepharose nor to concanavalin A-Sepharose. The chemotactic activity of NCF-Di was heat labile (56 C, 1 hr), but was resistant to periodate oxidation. These results suggest that NCF-Di is a simple peptide which has few or no sugar chains. These physicochemical properties of NCF-Di were compared to previously reported parasite-derived chemoattractants or purified allergen of D. immitis.

Defense reactions of mosquitoes to filarial worms: effect of host age on the immune response to Dirofilaria immitis microfilariae.

The melanization response of Aedes aegypti black-eyed Liverpool strain (LVP) and Aedes trivittatus against intrathoracically inoculated Dirofilaria immitis microfilariae (mff) was assessed in mosquitoes less than 1, 14, 21, and 28 days after adult ecdysis. There was a significant decrease in the melanization response of A. aegypti 14 days of age and older at 1, 3, and 5 days postinoculation (PI) compared to less than 1-day-old mosquitoes. The response also was reduced significantly in 14- to 28-day-old A. trivittatus on days 1 and 3 PI. Although essentially 100% of recovered mff were melanized by day 5 PI in A. trivittatus, the amount of melanin deposited was much less than that seen in 0-day-old mosquitoes. Potential mechanisms responsible for a reduced immune competence in older mosquitoes and the possible relationship to vector potential are discussed.

Prevalencia de Dirofilaria immitis en perros de Ciudad de La Habana

Se realizó el estudio de la prevalencia de Dirofilaria immitis en Ciudad de La Habana, mediante un muestreo al azar de 14 animales por municipio, para un total de 210 animales. Además, se analizó la posible relación del sexo, edad y condición del animal (callejero o casero), con la prevalencia de la enfermedad. Los resultados obtenidos fueron una prevalencia del 19(por ciento) para un total de 36 143 perros infestados en Ciudad de La Habana y una relación no significativa entre los factores analizados y la prevalencia de este parasitismo. Todos los datos fueron obtenidos y comprobados estadísticamente(AU)