Anaerobic fermentation results in loss of viability of Fasciola hepatica metacercariae in grass silage.

The parasitic liver fluke, Fasciola hepatica, has a detrimental impact on food security and poses a welfare concern to ruminant livestock. F. hepatica metacercariae, shed from an intermediate mud snail host, encyst on vegetation and present a source of infection to grazing livestock. Feeding grass silage to ruminants is a common practice, however the role it plays in the transmission of F. hepatica remains largely unknown. Our current understanding relies on historical studies that are not representative of current silage production and did not apply molecular methods to detect F. hepatica DNA persistence within silages. This study determined the impact of specific fermentation factors, including grass dry matter (DM) content (20, 30 & 40 %), length of ensiling period and maintaining an anaerobic environment on F. hepatica metacercariae viability. In vitro excystment assays demonstrated that regardless of grass DM content, metacercariae ensiled under anaerobic conditions were not viable from two weeks post-sealing. Metacercariae recovered from ensiled grass of 20 % DM content subjected to aerobic spoilage, remained viable for up to 10 weeks. DNA of F. hepatica remained detectable for up to 10 weeks in both anaerobic and spoiled silages. This study highlights i) the importance of maintaining an anaerobic ensiling environment to eliminate the risk of F. hepatica transmission from silage and ii) an inverse relationship between grass DM content and duration of metacercariae survival within spoiled silages. Improving our understanding of trematode metacercariae survival rates within silages, especially of highly pathogenic species such as F. hepatica, allows farmers to make informed decisions regarding on-farm parasite control.

Larval development sites of the main Culicoides species (Diptera: Ceratopogonidae) in northern Europe and distribution of coprophilic species larvae in Belgian pastures.

Some Culicoides species of biting midges (Diptera: Ceratopogonidae) are biological virus vectors worldwide and have indeed been associated with outbreaks of important epizoonoses in recent years, such as bluetongue and Schmallenberg disease in northern Europe. These diseases, which affect domestic and wild ruminants, have caused considerable economic losses. Knowledge of substrates suitable for Culicoides larval development is important, particularly for the main vector temperate species. This study, realized during two years, aimed to highlight the larval development sites of these biting midge species in the immediate surroundings of ten Belgian cattle farms. Moreover, spatial distribution of the coprophilic Culicoides larvae (C. chiopterus and C. dewulfi) within pastures was studied with increasing distance from farms along linear transects (farm-pasture-woodland). A total of 4347 adult specimens belonging to 13 Culicoides species were obtained by incubation of 2131 soil samples belonging to 102 different substrates; 18 of these substrates were suitable for larval development. The Obsoletus complex (formed by two species) was observed in a wide range of substrates, including silage residues, components of a chicken coop, dung adhering to walls inside stables, leftover feed along the feed bunk, a compost pile of sugar beet residues, soil of a livestock trampling area, and decaying wood, while the following served as substrates for the other specimens: C. chiopterus, mainly cow dung; C. dewulfi, cow dung and molehill soil; C. circumscriptus, algae; C. festivipennis, algae and soil in stagnant water; C. nubeculosus, algae and silt specifically from the edge of a pond; C. punctatus, mainly wet soil between silage reserves; C. salinarius, algae; and C. stigma, algae and wet soil between silage reserves. We also recorded significantly higher densities of coprophilic larvae within pastures in cow dung located near forests, which is likely due to the localization of potential hosts; the presence of these larvae within cow dung is, however, uninfluenced by relative distance from farms. A better knowledge of the microhabitats of Culicoides biting midges and their spatial distribution may allow the development of targeted species-specific vector control strategies, and may help to prevent the creation of new larval development sites.

Differential emergence of Culicoides (Diptera: Ceratopogonidae) from on-farm breeding substrates in Northern Ireland.

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are vectors of a number of viral diseases worldwide. Following the unforeseen outbreak of bluetongue in northern Europe (2006­2009) there was a need to clarify on-farm breeding substrates utilized by temperate Culicoides spp. Six substrates (cow dung, cow slurry, horse dung, sheep dung, maize silage and soil) were investigated for Culicoides spp. emergence over a 31-week period. Overall, most Obsoletus group Culicoides emerged from the cow dung and the most Pulicaris group Culicoides emerged from the sheep dung. Furthermore, Culicoides of the Obsoletus group were found to be abundant in cow slurry and sheep dung. Temperature played a significant role in the emergence times of adult Culicoides. The Obsoletus group appear to have undergone 3 generations during the experimental period. The sex ratio of emergent Obsoletus group Culicoides was affected by substrate type, with a greater proportion of males emerging from cow dung and slurry compared with the other substrates.

Chemical composition of silage residues sustaining the larval development of the Culicoides obsoletus/Culicoides scoticus species (Diptera: Ceratopogonidae).

Culicoides (Diptera: Ceratopogonidae) are biological vectors of bluetongue virus (BTV). Bluetongue is a viral disease that affects domestic and wild ruminants. Since its recent emergence in northern Europe, this disease has caused considerable economic losses to the sheep and cattle industry. The biotopes, and more particularly the chemical characteristics which are suitable for larval development of the main vector species, are still relatively unknown. This study shows that the larvae of biting midges belonging to the species Culicoides obsoletus and Culicoides scoticus are able to breed in different types of silage residue (maize, grass, sugar beet pulp and their combinations). The chemical composition of substrates strongly influences the presence of the immature stages of these biting midges. Higher lignin and insoluble fibre contents seem to favour their presence and could play the role of a physical support for semi-aquatic larvae. In contrast, higher concentrations of magnesium and calcium are negatively correlated with the presence of these two species. These data will help to locate and monitor the breeding sites of these species and could contribute to the control of these insects on farms.

Survival of model helminth eggs and larvae (Ascaris suum, Oesophagostomum sp.) in the ensilaging process.

Ascaris suum nonembryonated eggs remained viable for the most part even after 42 days of ensilaging. At the end of the anaerobic fermentation, mean of damaged eggs was 15.2 +/- 4.02 (min. 11, max. 21), 32.9%. Conversely, the viability of Oesophagostomum sp. nonembryonated eggs and infective L3 larvae was reduced-eggs: mean number 23.6 +/- 3.64 (min. 20. max. 28) specimens (93.3%), L3 larvae: mean number 24.2 +/- 4.38 (min. 19, max. 28) specimens (96.7%), during the period of study (42 days). Control group of the same helminth propagative stages, was kept under optimum aerobic conditions. After 42 days of exposition, 9.0 +/- 3.46 (min. 5, max. 11) nonembryonated Ascaris suum eggs (12.9%), 17.33 +/- 2.51 (min. 15, max. 20) Oesophagostomum sp. eggs (36.4%) and 3.66 +/- 1.15 (min. 3, max. 5) Oesophagostomum sp. larvae L3 (6.3%) were damaged on average. Helminth eggs, thick-walled and more resistant to the environment in particular, are able to survive the anaerobic process of ensilaging. To protect animals against parasitic diseases, it is necessary to consider the epidemiological hazard of silages and silage juices, which are potentially contaminated by helminth propagative stages. Silages and silage juices under certain conditions may become harmful to polygastric animals.