Ascaridia galli - An old problem that requires new solutions.

Reports of Ascaridia galli in laying hens in Europe have increased since the ban on conventional battery cages in 2012. As this parasite is transmitted directly via the faecal-oral route by parasite eggs containing a larva, it is reasonable to assume that the escalating problem is related to the increased exposure now occurring in modern welfare-friendly cage-free housing systems. On many farms, A. galli reappears in subsequent flocks, even though the birds have no access to the outdoors, biosecurity is high and empty houses are cleaned and disinfected during downtime. Since the egg production cycle lasts only ≈80 weeks and recombinant antigen production for helminth vaccines has not yet been solved, the development of a vaccine seems to be an unrealistic option. Therefore, disrupting the life cycle of the parasite by other means, including the strategic use of dewormers, appears to be the key to controlling infection. Of concern is that only one class of anthelmintics is licenced for poultry in Europe and that are usually administered indiscriminately through the birds' drinking water and often too late when the parasite is already established. If current calendar-based parasite control strategies are not changed, there is a risk that resistance to anthelmintics may develop, as has already been demonstrated with nematodes in livestock. We insist that treatments can be more effective and the risk of developing drug resistance can be mitigated if we invest in a better understanding of A. galli responses to more prudent and judicious use of anthelmintics. This review identifies knowledge gaps and highlights aspects of sustainable parasite control that require further research to support commercial egg producers.

Implementation of a targeted treatment strategy for the sustainable control of Ascaridia galli infections in laying hens.

Background: Ascaridia galli is a widespread problem in cage-free egg production. Sustainable control of nematode infections is a key component in this sector. This study investigates the effect of a treatment strategy against A. galli, aiming to propose a guideline for anthelmintic use on commercial poultry farms. Methods: A total of eight flocks of laying hens (a-h) from five commercial poultry farms were included in this study. Faecal samples were collected on a biweekly basis starting at 7-13 weeks post-placement (WPP) and processed using the McMaster method to calculate ascarid egg shedding. Flocks were treated after the threshold of 200 eggs per gram of faeces (EPG) was reached. Results: The highest initial faecal egg count was 6700 EPG at 11 WPP, whereas the lowest was 50 at 8 WPP. The longest delay to detect A. galli was 7 weeks. The lowest and the highest number of treatments were four and six, respectively. The shortest and longest periods between any two treatments were 5 and 22 weeks, respectively. Conclusions: These results suggest that monitoring for A. galli should start at approximately 7 WPP and should be repeated every 8 weeks until hens are 50 weeks old. Treatment should be given only if moderate to high faecal egg counts are observed. Treatments after this point may be repeated every 8 weeks without eventually performing a faecal test. These findings provide practical support to veterinarians and egg producers dealing with ascarid worm infection in laying hens in their production stage.

RNA-Seq de novo assembly and differential transcriptome analysis of the nematode Ascaridia galli in relation to in vivo exposure to flubendazole.

The nematode Ascaridia galli (order Ascaridida) is an economically important intestinal parasite responsible for increased food consumption, reduced performance and elevated mortality in commercial poultry production. This roundworm is an emerging problem in several European countries on farms with laying hens, as a consequence of the recent European Union (EU) ban on conventional battery cages. As infection is associated with slow development of low levels of acquired protective immunity, parasite control relies on repeated use of dewormers (anthelmintics). Benzimidazoles (BZ) are currently the only anthelmintic registered in the EU for use in controlling A. galli and there is an obvious risk of overuse of one drug class, selecting for resistance. Thus we developed a reference transcriptome of A. galli to investigate the response in gene expression before and after exposure to the BZ drug flubendazole (FLBZ). Transcriptional variations between treated and untreated A. galli showed that transcripts annotated as mitochondrial glutamate dehydrogenase and cytochrome P450 were significantly down-regulated in treated worms, whereas transcripts homologous to heat shock proteins (HSP), catalase, phosphofructokinase, and a multidrug resistance P-glycoprotein (PGP1) were significantly up-regulated in treated worms. Investigation of candidate transcripts responsible for anthelmintic resistance in livestock nematodes led to identification of several tubulins, including six new isoforms of beta-tubulin, and several ligand-gated ionotropic receptors and ABC-transporters. We discovered several transcripts associated with drug binding and processing genes, but further characterisation using a larger set of worms exposed to BZs in functional assays is required to determine how these are involved in drug binding and metabolism.

Environmental tolerance of free-living stages of the poultry roundworm Ascaridia galli.

The poultry roundworm Ascaridia galli is re-emerging in laying hens in many European countries due to the increase in non-caged housing. A series of in vitro experiments was carried out to study the in ovo larval development (embryonation) under different environmental conditions. Between 83% and 96% of the eggs developed to L3 within 7-21 days of incubation in water between 20 and 30°C. Twenty-six percent completed development at 33°C and 4% at 35°C after 31 days. At 15°C parasite egg development was low with 8% L3 after 56 days. In another trial larval development occurred, when parasite eggs were exposed to freeze-thaw cycle (30' to 12h) followed by incubation for 2 weeks at 25°C. Alkaline and acidic conditions in the range of pH 2.5-12.5 had no adverse effect on development. Oxygen and relative humidity above 70% were necessary for development to occur. Thus, some A. galli eggs may complete development at conditions prevailing in poultry barns in temperate climate zones throughout the year. Although exposure to a 1% or 2% dilution of the broad-spectrum disinfectant chlorocresol for 4h or longer was ovicidal, further work is required to improve the method of application in the field.