Immunoproteomic Analysis of Dirofilaria repens Microfilariae and Adult Parasite Stages

Dirofilariarepens is a parasitic nematode causing a vector-borne zoonotic infection (dirofilariosis), considered an emerging problem in human and veterinary medicine. Currently, diagnosis is based on the detection of the adult parasite and microfilariae in the host tissues. However, the efficacy of tests relying on microfilariae detection is limited by microfilariae periodic occurrence. Therefore, a new reliable and affordable serological diagnostic method is needed. Better characteristic of the parasite biology and its interaction with host immune system should help to achieve this goal. This study analyzes adult and microfilariae proteomes, and the use of one-dimensional electrophoresis (1-DE) and two-dimensional electrophoresis (2-DE) proteomics, immunoproteomics, and LC-MS/MS mass spectrometry allowed us to identify 316 potentially immunogenic proteins (75 belong to adult stage, 183 to microfilariae, and 58 are common for both). Classified by their ontology, the proteins showed important similarities and differences between both parasite stages. The most frequently identified proteins are structural, metabolic, and heat shock proteins. Additionally, real-time PCR analysis of some immunogenic targets revealed significant differences between microfilariae and adult life stages. We indicated molecules involved in parasite-host interactions and discussed their importance in parasite biology, which may help to reveal potential diagnostic antigens or select drug and vaccine targets.

Methods for quantifying eggs and oviposition rate of Dawestrema cycloancistrium (Monogenea: Dactylogyridae), monogenean parasite of Arapaima gigas (Teleostei: Osteoglossidae)

Dawestrema cycloancistrium is the main ectoparasite causing mortality in fingerlings of pirarucu (Arapaima gigas) in Amazonian aquaculture. Very little is known about the D. cycloancistrium reproductive index and appropriate methods of collecting eggs for investigation. This study aimed to determine the oviposition rate of D. cycloancistrium. To achieve this aim, two egg quantification methods were tested: the estimative method (ME) and the total counting method (MT). Compared with the MT, the ME overestimated the number of eggs counted, which were 2943.5 ± 2840.6 and 1041.5 ± 533 eggs, and the oviposition rate, which was 80.1 ± 58.7 and 31.4 ± 16.4 eggs/parasite/day, for ME and MT, respectively. These results show that for studies quantifying D. cycloancistrium eggs, the total eggs in the sample must be counted, as the estimates made using subsamples are not representative. Using the MT, the oviposition rate for D. cycloancistrium was determined to be 31.4 ± 16.4 eggs per adult parasite per day. The present study demonstrates the egg production capacity of a monogenean species parasite of A. gigas, providing basic biological data for D. cycloancistrium.

Modelling the effect of nematode parasites on the liveweight gain of lambs grazing alternately with cattle

A dynamic model for nematode parasites in lambs which links their adult parasite burden with liveweight gain was extended to investigate alternate grazing with cattle to provide relatively larval-free pasture for finishing lambs. Liveweight performance of weaned lambs was tracked fortnightly under rotational grazing using three cattle:sheep ratios, with three initial parasite burdens and two initial infective larval densities on pasture. Lambs were removed from grazing at 37 kg or if they failed to gain weight during the preceding fortnight. An 80%:20% cattle:sheep ratio reduced larval pasture contamination by 75% but did not affect parasitic burden and total lamb liveweight gain compared with an all-lamb treatment. For low and moderate initial parasitic burdens in the 80%:20% cattle:sheep ratio, 25% of lambs reached 37 kg and 100% gained weight until week 18. This simulation indicated that in organic lamb finishing systems, alternate grazing with cattle provides substantial benefit but only when the initial parasite burden was low or moderate. Keywords: modelling, parasites, liveweight gain, alternate grazing