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Climate change, competent vectors, and reservoir animals are the main factors for developing vector-borne zoonotic diseases. These diseases encompass a significant and widespread category of pathogens (e.g., viruses, bacteria, protozoa, and helminths) transmitted by blood-feeding arthropods, including ticks, fleas, lice, triatomines, mosquitoes, sandflies, and blackflies. In Chile, several studies have explored the role of dogs as reservoirs of vector-borne pathogens; however, there is a lack of research investigating the presence of pathogens in arthropods. Specifically, within the order Diptera, limited knowledge exists regarding their roles as carriers of pathogens. This study aimed to examine the presence of zoonotic filarial nematodes in mosquitoes and dogs within a previously unstudied semi-rural area of Central Chile. Two hundred samples of dog blood and seven hundred and twenty-four mosquitoes were collected during 2021–2022 and studied for filarial nematodes by PCR. The prevalence of microfilaremic dogs detected by Knott’s test was 7.5%, with Acanthocheilonema reconditum being the only species identified. Aedes ( Ochlerotatus ) albifasciatus was the most abundant mosquito species collected, and 15 out of 65 pools were positive for filarial nematodes. Among these pools, 13 tested positive for Acanthocheilonema reconditum , and two tested positive for Setaria equina through PCR. Additionally, five Culex pipiens specimens were positive for Acanthocheilonema reconditum . Despite the absence of zoonotic filarial species, these findings underscore the significance of monitoring pathogens in mosquitoes and animal hosts and continued research into the dynamics of vector-borne diseases, particularly in unexplored regions.

Iran is one of the endemic areas of Dirofilariasis, and also one of the most important zoonotic infections. Dirofilaria immitis causes a severe and fatal disease called heartworm disease in dog. It also produces pulmonary nodules in humans. The worm is to be investigated as a potential infection of humans and animals in various provinces in Iran. In this research, the samples were studied with modified Knott’s test and molecular method. The results of the modified Knott’s test method indicated that 14.00% of sheepdogs were infected with filarial microfiler. The microfilers were characterized with basic morphological features, the length of the infective larva and tail ending. There was an estimated prevalence of 4.45% for Dirofilaria immitis and 9.55% for Acanthocheilonema reconditum microfiler. To verify the differential diagnosis, molecular method was performed using PCR with Dirofilaria specific primers for amplification of ITS2 locus. Gene locus sequencing results of D. immitis and sequence alignment recorded in GeneBank showed 97.00% similarity, and relatively 98.00% similarity was observed in A. reconditum. The results of the molecular method confirmed the result of modified Knott’s test method. Low infection with D. immitis was observed the region, probably due to the fact that the annual temperature and precipitation in this area were not suitable for the proliferation of the vector mosquitoes. In general, there was less infection in the region compared to regions with relatively similar climatic conditions. Hence, the results suggested that alternative diagnostic tests are required to determine the occult infections.