Porcisia transmission by prediuresis of sand flies

Abstract

Parasites of the genus Porcisia, together with the genus Endotrypanum, form a sister clade to the species-rich and medically important genus Leishmania. Both Porcisia species, P. hertigi and P. deanei, are dixenous parasites of Neotropical porcupines. Almost 50 years after their first discovery, knowledge of their life cycle remains poor and their insect vectors are unknown. Because competent vectors of their closest phylogenetic relatives, genera Endotrypanum and Leishmania, are phlebotomine sand flies (Diptera: Psychodidae) and/or biting midges (Diptera: Ceratopogonidae), we examined here the potential of both sand flies and biting midges to transmit Porcisia parasites. The insects (Lutzomyia longipalpis, L. migonei and Culicoides sonorensis) were exposed to parasites through the chicken skin membrane and dissected at various time intervals post bloodmeal. Potentially infected females were also allowed to feed on the ears of anaesthetized BALB/c mice and the presence of parasite DNA was subsequently confirmed in the mice by PCR. Porcisia hertigi did not survive defecation in L. longipalpis or L. migonei, suggesting that these sand fly species are unlikely to serve as natural vectors of this parasite. Similarly, P. hertigi infections were lost in Culicoides midges. In contrast, mature P. deanei infections developed in 51-61% of L. longipalpis females, 7.3% of L. migonei females and 7.7% of Culicoides sonorensis females. In all three vector species, P. deanei colonized predominantly Malpighian tubules and produced metacyclic infective forms. Transmission of P. daenei to BALB/c mice was demonstrated via the prediuresis of L. longipalpis females. This mode of transmission, as well the colonization of Malpighian tubules as the dominant tissue of the vector, is unique among trypanosomatids. In conclusion, we demonstrated the vector competence of L. longipalpis for P. deanei but not for P. hertigi, and further studies are needed to evaluate competence of other Neotropical vectors for these neglected parasites.

Leishmaniinae: evolutionary inferences based on protein expression profiles (PhyloQuant) congruent with phylogenetic relationships amongLeishmania, Endotrypanum, Porcisia, Zelonia, Crithidia,andLeptomonas

“…Intracellular amastigotes are obligatory in Leishmania and have been recorded for E. colombiensis in man. Porcisia in porcupines and Endotrypanum in sloths and squirrels apparently do not cause any disease symptoms [4, 13–17].…”

“…The species of Porcisia , so far exclusive to Neotropical porcupines, form together with the genus Endotrypanum a clade sister to Leishmania . The life cycle of Porcisia remains unclear, but a recent experimental study demonstrated an unexpected contaminative transmission of P. deanei by both sand flies and biting midges [17]. Zelonia are apparently a monoxenous genus consistently placed in the major clade nesting all dixenous species.…”

Steppe lemmings and Chinese hamsters as new potential animal models for the study of the leishmania subgenusMundinia(Kinetoplastida: Trypanosomatidae)

“…Total DNA isolated using the High Pure PCR Template Preparation Kit (Roche Diagnostics, Indianapolis, IN) according to the manufacturer’s instructions was used as a template for nested PCR amplification (detection limit 10 -2 parasites per sample) with outer primers amplifying 332 bp region of 18S sequence (forward primer 18SN1F 5’-GGA TAA CAA AGG AGC AGC CTC TA3’ and reverse primer 18SN1R 5’– CTC CAC ACT TTG GTT CTT GAT TGA-3’) and inner primers amplifying the 226 bp long 18S sequence (forward primer 18SN2F 5′-AGA TTA TGG AGC TGT GCG ACA A-3′ and reverse primer 18SN2R 5′-TAG TTC GTC TTG GTG CGG TC-3′) previously used by Sadlova et al 2022 (38). The samples were then analysed using 0,8% agarose gel.…”

Artificial Feeding Systems for Vector-Borne Disease Studies

“…Capillary Feeding : More specialized than membrane feeding devices are capillary feeding systems, offering an alternative artificial feeding approach specifically designed for certain hematophagous insects [ 146 ]. In this system, vectors like ticks feed through a capillary tube filled with a blood substitute or animal/human blood.…”

“…This has also been employed to assess the transmission of Leishmania spp. in sandflies [ 150 ] and investigate the vector competence of sandflies [ 146 ].…”