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Backyard chickens usually have open housing and scavenge for food, facilitating contact with infective forms of parasites present in the environment and increasing the risk of parasitic infections. Coccidiosis and ascaridiosis are two internal parasites in scavenging chickens that cause production losses to farmers. Prevalence data of these two diseases are scarce in low-income and middle-income countries (LMICs) but necessary to assess the burden of disease in backyard chickens. This study estimates the clinical prevalence of coccidiosis and ascaridiosis at the country and regional levels in all LMICs using regression imputation methods. Regression models were developed with data of known prevalence of these parasites and using primarily climatic factors as predictors. A binomial random effects model was developed for each disease to impute prevalence. The overall estimated prevalence of coccidiosis was 0.39 (95 % CI: 0.37-0.42), with a higher mean prevalence in tropical countries. The overall estimated prevalence of ascaridiosis was 0.23 (95 % CI: 0.22-0.25) with a higher mean prevalence in countries with tropical, temperate and continental climates. The findings of this study can aid to identify the burdens of coccidiosis and ascaridiosis infections across countries and regions, which can inform disease control plans and, and encourage international alliances to facilitate access to preventive measures, improving animal health and reducing production losses., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Chicken production has increased over the past decade, resulting in a concomitant rise in the demand for more humane options for poultry products including cage-free, free-range, and organic meat and eggs. These husbandry changes, however, have come hand-in-hand with increased prevalence of Ascaridia galli infection, which can cause clinical disease in chickens as well as the occasional appearance of worms in eggs. Additionally, development of anthelmintic resistance in closely related helminths of turkeys highlights the need for closely monitored anthelmintic treatment programs. Manual faecal egg counts (FECs) can be time-consuming and require specialist training. As such, this study sought to validate an automated FEC system for use in detection and quantification of A. galli eggs in chicken faeces. Automated counts using the Parasight System (PS) were compared to traditional manual McMaster counting for both precision and correlation between methods. Overall, ten repeated counts were performed on twenty individual samples for a total of 200 counts performed for each method. A strong, statistically significant correlation was found between methods (R 2 = 0.7879, P < 0.0001), and PS counted more eggs and performed with statistically significant higher precision ( P = 0.0391) than manual McMaster counting. This study suggests that PS is a good alternative method for performing A. galli FECs and provides a new tool for use in helminth treatment and control programs in chicken operations.

Anthelmintic resistance in livestock animals has been spreading across the world in prevalence and severity. As a result, researchers are exploring alternative strategies to combat this issue, and one promising avenue is the utilization of medicinal plants. This study aims to investigate the anthelmintic efficacy of the crude ethanolic extract (CEE) derived from the leaves of Juglans regia against one of the most detrimental nematode parasites affecting poultry, namely Ascaridia galli (A. galli). For the in vitro studies, adult A. galli worms were collected from the naturally infected chickens and the efficacy of CEE was measured at the concentration of 25, 50, and 100 mg/ml using adult worm motility inhibition (WMI) assay. In addition, levamisole (0.55 mg/ml) was used as the positive control. Likewise, Phosphate buffered saline (PBS) was used as the negative control. For the in vivo studies, CEE of J.regia at the doses of 500, 1000, and 2000 mg/kg were evaluated in chickens experimentally infected with A. galli. The anthelmintic efficacy was monitored using faecal egg count reduction (FECR) and worm count reduction (WCR) assays. In vitro studies revealed significant (P < 0.001) anthelmintic effects of CEE of J.regia on the motility of A. galli worms at different hours post-exposure. At the concentration of 100 mg/ml, CEE resulted in 96.5% inhibition of worm motility at 24 h post-exposure. While the synthetic anthelmintic drug, levamisole caused the highest inhibition of worm motility (100%) at the same time period. The in vivo anthelmintic activity of CEE of J. regia demonstrated a maximum effect on day 14 post-treatment by inducing 67.28% FECR and 65.03% WCR. We observed no significant difference (P > 0.05) in worm counts between the negative control group and the chickens treated with CEE at the dosage of 500 mg/kg. Together, the results of the present study suggest that CEE of J. regia leaves possess anthelmintic properties and could be a potential source of novel anthelmintic compounds for controlling helminth parasites., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Raillietina species and Ascaridia galli are two of the significant intestinal parasites that affect chickens in a free-range system production. They destroy the intestinal mucosa layer, leading to several clinical symptoms such as weight loss, a slowed growth rate, and economic value loss. Thus, the objective of this study was to develop an assay for simultaneously detecting Raillietina spp. (R. echinobothrida, R. tetragona, and R. cesticillus) and A. galli in a single reaction using duplex loop-mediated isothermal amplification (dLAMP) coupled with a lateral flow dipstick (LFD) assay. The analytical specificity of the dLAMP-LFD assay showed a high specific amplification of Raillietina spp. and A. galli without non-target amplification. Regarding the analytical sensitivity, this approach was capable of simultaneously detecting concentrations as low as 5 pg/μL of mixed-targets. To evaluate the efficiency of the dLAMP assay, 30 faecal samples of chickens were verified and compared through microscopic examination. The dLAMP-LFD assay and microscopic examination results showed kappa values of Raillietina spp. and A. galli with moderate (K= 0.615) to high (K= 1) agreements, respectively, while the McNemar's test indicated that the efficiency between assays was not significantly different. Therefore, the developed dLAMP-LFD assay can be used as an alternative screening method to the existing classical method for epidemiological investigation, epidemic control, and farm management, as well as for addressing poultry health problems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Aim of the Study: The growing resistance of helminth parasites to currently available commercial anthelmintic drugs, combined with apprehensions regarding detrimental chemical residues in livestock products, has sparked an interest in exploring medicinal plants as an alternative strategy for treating helminthiasis. As a result, this study was designed to investigate the anthelmintic activity of crude methanolic extracts (CME) of Saussurea costus root on Ascaridia.galli, a pathogenic nematode of poultry., Materials and Methods: In vitro, the anthelmintic effect of Saussurea costus root was evaluated in comparison to commercial anthelmintic, levamisole on the adult nematode parasites, A.galli using worm motility inhibition (WMI) test. The CME of S.costus was also evaluated for in vivo anthelmintic activity in chickens experimentally infected with Ascaridia galli. For the in vivo study, one hundred-day-old chickens were orally infected with embryonated eggs of A. galli worms. The efficacy of the plant extract as an anthelmintic was assessed through two tests: faecal egg count reduction (FECR) test and worm count reduction (WCR) test. The study investigated three distinct doses of plant extract under in vivo setup: 500 mg kg -1 body weight (bw), 1000 mg kg -1 bw, and 2000 mg kg -1 bw., Results: In vitro, all the tested concentrations of S.costus (25 mg/ml, 50 mg/ml, and 100 mg/ml) showed a significant (P < 0.001) anthelmintic effects on live adult A. galli worms in terms of inhibition of worm motility at different hours post-treatment. At the highest concentration of the extract, we observed worm motility inhibition of 100% at 24 h post-exposure. On day 14 post-treatment, all birds were slaughtered, and adult A. galli worms were subsequently retrieved from their small intestines. Birds treated with CME extract of S. costus root exhibited a significant (P < 0.001) reduction in faecal egg count. However, the administration of the extract at the dosage of 500 mg kg -1 bw to the birds did not reveal any significant (P > 0.05) differences in the worm count compared to the negative control group. The CME of S. costus at a dose of 2000 mg kg -1 bw showed the highest anthelmintic activity by inducing 83.10% FECR and 76.47% WCR., Conclusion: In conclusion, the root extract of S. costus has a promising anthelmintic activity on A. galli as demonstrated by the results of the present experiment., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Ascaridia galli is the largest gut-dwelling helminth of chickens, which confers adverse effects on meat and egg production; thus, on the animal protein supply and the economy. Both adult and immature parasites affect gut health, but larval stages play a major role in pathology., Aims: Here, we present immunology and pathology of A. galli in chickens., Materials and Methods: Literatures were surveyed through online platforms such as PubMed, Google Scholar and Researchgate., Results: The larvae cause excessive mucus production, damage to the intestinal gland, hemorrhage, anemia, diarrhea, and malnutrition. The adult worms can cause death by intestinal obstruction and intussusception. Although both cellular and humoral immunity are involved in fighting against ascariasis, the role of naturally acquired immunity is poorly defined. In cellular immunity, Th-2 cytokines (IL-4, IL-5, IL-9, and IL-13), goblet cells (mucin), gut-associated lymphoid tissues, CD8α+ intraepithelial cells, TCRγδ + T cells, and TGF-β4 form a protective band. Type 2 immunity provides protection by forming a network of endogenous damage-associated molecular patterns, chitin, and parasitic antigens. Among antibodies, IgY is the most prominent in chickens and provides temporary humoral protection. During parasitic infection, infiltration of various immune cells is evident, especially in the intestinal epithelium, lamina propria, and crypts of the duodenum and jejunum. In chickens older than 12 weeks, gradual reduction of worm burden is more successful than the younger birds. Female chickens exert a short-lived but higher level of protection by passing IgY to chicks in the form of egg yolk antibodies. In laying conditions, immunity differs between breeds. This review provides an overview of the silent but inevitable pathological changes induced by A. galli and the interaction of host immunity with the parasite., (© 2023 The Authors. Immunity, Inflammation and Disease published by John Wiley & Sons Ltd.)

Chicken production is quickly rising due to the low associated costs and the capability of poultry to convert nutrients into biological protein along with chicken meat accounting for 30% of all animal protein eaten by humans. Despite advances in poultry production, parasitic illnesses in laying hens remain a problem. Farm birds reared in semi-intensive and free-range systems are more prone to parasite infections due to the absorption of polluted water and food from scavenging behaviors and waste droppings. In this study, the effects of Ascaridia galli infection on the immune response and liver function of two laying hen lines are compared, and their infection resistance is determined. In total, 50 laying hens at eight weeks of age were used (25 Lohmann brown-classic and 25 Lohmann lsl-lite), and each line was divided into two groups: an infected group (n=15), which was orally infected with a single dose of 500 A. galli embryonated eggs, and a control group (n=10), which was given normal saline. After four and eight weeks, blood was collected from the wing vein to assess the serum's AST, ALT, total protein, and IgY levels. The results demonstrated that the infected Lohmann brown-classic and Lohmann lsl-lite chickens presented significantly increased (P≤0.05) AST, ALT, and IgY, compared to the respective control group. Moreover, Lohmann brown-classic hens presented a significantly increased (P≤0.05) IgY concentration four weeks after infection, compared to Lohmann lsl-lite hens. From our results, it can be concluded that genetic variation plays a crucial role in the immune response against A. galli , where the Lohmann brown-classic line was found to be more resistant, compared to the Lohmann lsl-lite line., Competing Interests: The authors declare that they have no conflict of interest.

The present study was performed to evaluate the in vivo efficiency of Curcurbita pepo (pumpkin) seeds, Cymbopogon citratus (lemongrass) essential oil and Plantago lanceolata (ripleaf) leaves against helminth infections in laying hens. In the first experiment, 75 Lohmann LSL Classic hens naturally infected with Ascaridia galli were assigned to groups of five; groups were randomly assigned to one of three treatments with five replicates each (untreated control; lemongrass oil: 1 g/bird/day; pumpkin seeds: 10 g/bird/day). Feed consumption and egg production were continuously recorded, individual faecal egg counts were determined weekly, and E. coli and Lactobacillus spp. three times during the experimental period of 29 days. After slaughter, intestinal worms were counted and sexed. Pumpkin improved feed conversion as compared to the control (p = 0.008) and to lemongrass (p = 0.021); no treatment effect on any other parameter was found. In the second experiment, 75 LSL pullets were artificially infected with 3 × 200 A. galli eggs, randomly divided into groups of five and assigned to one of three treatments (untreated control, lemongrass oil: 1 g/bird/day; ripleaf: 5% of ration). After 109 days of sampling as described above, hens were slaughtered and worm burdens determined. Performance of the animals did not change regardless of the treatment and none of the treatments resulted in changes of the microbiological and parasitological parameters. In conclusion, with the exception of improved feed conversion in the pumpkin group, no positive nor negative effects of the additives on performance, parasitological and microbiological parameters of naturally and artificially A. galli infected laying hens were observed., (© 2022. The Author(s).)

This study investigated the pharmacokinetics and therapeutic efficacy of levamisole (LVS) after intravenous (i.v.) and oral administrations to healthy and Ascaridia galli-infected ducks by developing an infection model. Twenty-four two-week old ducklings were experimentally infected with A. galli. The ducks were monitored for the development of infection and after 8 weeks they were administered with LVS at a single dose of 30 mg/kg by oral or i.v. administration. Sixteen healthy ducks were subjected to the same treatment and served as control. Serial blood samples were taken for LVS determination with HPLC-UV and pharmacokinetic analysis was carried out based on the non-compartmental approach. The LVS therapeutic efficacy was determined 1 week post drug administration by intestinal worm count at necropsy. In vivo data on development of ascariasis in ducks showed that 8 weeks post inoculation the number of eggs per gram of feces reached at least 100 in each bird. After a single dose of LVS, no parasites were recovered upon necropsy. Results of the pharmacokinetic study showed no statistical differences between infected and non-infected birds for both routes of administration. The mean oral bioavailability was slightly below 50% in both experimental groups. In conclusion, the pharmacokinetics of LVS in ducks was not affected by experimentally-induced ascariasis. A single dose of LVS was found to be efficient against experimental ascariasis in ducks induced by in field isolates of A. galli., Competing Interests: Declaration of Interest Authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

A reliable method of diagnosing the most prevalent helminth infections in chickens is vital for developing effective control strategies. Ascaridia galli and Heterakisgallinarum are phylogenetically close nematode species that can elicit the development of cross-reactive antibodies in chickens. Therefore, an enzyme-linked immunoassay (ELISA) based on Ascaridia galli antigens in faeces of chickens to detect and quantify infections with both A. galli and H. gallinarum was developed. The ELISA utilised polyclonal antibodies that were obtained from rabbits immunised with soluble antigens isolated from A. galli. In two separate experiments, chickens were kept as uninfected controls or were orally infected with either 100 or 1000 of embryonated eggs of A. galli or H.gallinarum. Faecal samples were collected after 28-30 weeks post-infection. The ELISA was then used to quantify the concentration of soluble worm antigens in faecal samples, i.e., the amount of antigen per gram faeces, APG. The APG from infected chickens was significantly higher than non-infected groups in both experiments (P 0.001). Both 100 and 1000 infection dose groups were not significantly different (P = 0.999) in the experiment with H. gallinarum, whereas in the experiment with A. galli, APG was significantly higher in the 1000 infection group (P 0.001). A receiver operation characteristics (ROC) analysis that evaluates the qualitative performance of diagnostics tests was used to calculate the assay parameters within each mono-infection experiment. The result showed that the assay had a high diagnostics accuracy with an area-under-curve (AUC) of 0.99 in detecting infection in A. galli infected chickens and a moderate-high accuracy (AUC = 0.89) in birds infected with H. gallinarum. The diagnostic sensitivity and specificity of the assay at the optimal cut-off point equivalent to Youden index were 93% and 100% for detecting infections in A. galli experiment and 85% and 92% in H. gallinarum experiment, respectively. The correlation between faecal antigen concentration and all worm burden parameters was positive but generally low (r < 0.33), which provided less information about infection intensities. Nonetheless, these results indicate that a reliable and accurate qualitative diagnosis of the two most prevalent intestinal nematodes in chickens can be achieved using a non-invasive copro-antigen ELISA assay., Competing Interests: Conflict of interest The authors have read the journal’s policy and have the following competing interests: MH is an employee of TECODevelopment GmbH. This does not alter our adherence to Veterinary Parasitology policies on sharing data and materials. There is no patent associated with this research to declare. Other authors have no competing interest., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Ascaridia galli infection models use eggs isolated from chicken excreta, worm uteri and worms cultured in artificial media. The aim of this study was to compare the infectivity of A. galli eggs isolated from these sources under two infection regimens. A 3 × 2 factorial arrangement was employed to test the infectivity of A. galli eggs from the three sources and two modes of infection (single or trickle infection). One hundred and fifty-six Isa-Brown one day-old cockerels randomly assigned to the six treatment groups (n = 26) were orally infected with embryonated A. galli eggs obtained from the three A. galli egg sources (worm uteri, excreta or eggs shed in vitro) administered either as single dose of 300 eggs at one day-old or trickle infected with 3 doses of 100 eggs over the first week of life. Twenty-two negative control birds remained uninfected. Eggs obtained from cultured worms or excreta exhibited a higher embryonation capacity (P = 0.003) than eggs obtained from worm uteri. There were higher worm establishment (infectivity) rates from embryonated eggs originating from cultured worms and worm uteri compared with eggs obtained from fresh excreta (P < 0.0001). Trickle infection resulted in a significantly higher total worm burden (P = 0.002), establishment rate (P = 0.002) and excreta egg counts (EEC, P = 0.025) than single infection. Worm length was greater in birds infected with embryonated eggs from excreta than from uteri or cultured worms (P < 0.0001). However, mode of infection did not affect worm length (P = 0.719) and weight (P = 0.945). A strong significant positive linear correlation was observed between EECs and female worm counts at 12 weeks of post infection sampling (r = 0.75; P < 0.0001). Body weight of birds was negatively correlated with both worm burden (r = - 0.21; P < 0.01) and EEC (r = - 0.20; P < 0.05) at 12 weeks post infection. In conclusion, our results show that eggs shed by cultured worms or isolated from worm uteri had greater infective capacity than eggs harvested from excreta and that trickle rather than bolus infection resulted in higher worm establishment. These factors should be taken into account when considering artificial infection protocols for A. galli., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Eggs in the infective stage of the chicken nematode Ascaridia galli are often required for in vivo and in vitro studies on this parasite. The reliability of any artificial A. galli infection depends on the viability and embryonation capacity of A. galli eggs. The aim of this study was to determine ideal storage conditions for maximising the viability of A. galli eggs and maintaining viability for the longest period. A 2 × 2 × 3 × 5 factorial experimental design was employed to investigate the effects of storage temperature (4°C or 26°C), storage condition (aerobic or anaerobic), storage medium (water, 0.1 N H 2 SO 4 or 2% formalin) and storage period (4, 8, 12, 16 and 20 weeks). The viability of eggs was assessed after eggs in all treatment groups were held aerobically at 26°C for 2 weeks after the storage period to test embryonation capacity. Based on morphological characteristics, they were categorised as undeveloped, developing, vermiform, embryonated or dead. The maintenance of viability during storage at 4°C was optimal under anaerobic conditions while at 26°C it was optimal under aerobic conditions. Anaerobic conditions at 26°C led to a rapid loss of viability while aerobic conditions at 4°C had a less severe negative effect on maintenance of viability. Egg storage in 0.1 N H 2 SO 4 resulted in a significantly higher viability overall (54.7%) than storage in 2% formalin (49.2%) or water (37.3%) (P < 0.0001). Untreated water was the least favourable storage medium when eggs were stored at 26°C while it was a medium of intermediate quality at 4°C. The viability of A. galli eggs decreased significantly with storage time (P < 0.0001) depending on the other factors. The lowest rate of decline was seen with storage of eggs under anaerobic conditions at 4°C or aerobic conditions at 26°C in 0.1 N H 2 SO4. Eggs in these treatments retained up to 72% of overall viability at 20 weeks with a decline rate of approximately 2% per week with no significant difference between the two. Therefore, this study has clearly revealed opposing aerobic conditions required for prolonged storage of A. galli eggs in the pre-embryonated state at 4°C. It has also identified that 0.1 N H 2 SO4 provides the best preservation against degradation during storage, particularly at 26°C under aerobic conditions. Achieving strictly anaerobic conditions can be difficult to achieve so storage aerobically at 26°C may be preferred for simplicity., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Because of the trend of cage-free egg production, infections with the nematode Ascaridia galli are receiving increased attention. The aim of this study was to establish a timeline for the influence of A. galli on the expression of key cytokines related to a parasitic immune response, and on the composition of the jejunal microbiota. Twenty-eight male layer-type birds were challenged at 24, 25, and 26 days of age. An additional 28 birds were kept as uninfected controls. Starting on Day 31, three birds of each group were euthanized every week until 8 wk postinfection (PI). The number of larvae isolated from the intestinal wall decreased over time, until no larvae were seen at 7 and 8 wk PI. At 5 wk PI, there was a numerical upregulation of all cytokines (TGF-β, IFN-γ, IL-4, IL-8, IL-10, IL-13) in the infected group, but this change was only statistically significant for IL-13. At this time point, larvae were expected to have developed into adults that would have shed eggs in the feces. However, no adult worms were seen and there was no egg shedding. For the microbiota analysis, there were significant differences in the alpha diversity (Faith's phylogenetic diversity) between challenge and control groups, and the beta diversity analysis showed slight differences between samples, suggesting that the age of the birds was the main reason for the separation of groups. These findings suggest that the upregulation of all cytokines evaluated in Week 5 might be the reason for resolution of the infection. Possible explanations are that a high infection dose and the fact that birds were fed with a more nutritionally dense feed might have contributed to the birds' immune system clearing the infection before the worms were able to reach maturity.

Parasitism is a divesting problem that is frequently overlooked and may result in severe prominent clinical manifestation. This study aimed to investigate the seasonal and sexual prevalence of the gastrointestinal nematode Ascaridia columbae (A. columbae) infection among domestic pigeons in Giza governorate, Egypt, during the period from 2020 to 2021. One hundred and sixty suspected pigeons were clinically investigated. Blood & tissue samples were collected from infected birds to estimate serum zinc concentration, malondialdehyde (MDA), and nitric oxide levels. As well as tumor necrosis factor-alpha (TNF-α), interleukin 1β (IL1β) activity, and histopathological examination were estimated; also, worms were collected for morphological identification using electron microscope (SEM) and molecularly identified using polymerase chain reaction (PCR), further sequenced, and submitted in GenBank with accession number MZ343369. The average ascarid (length × breadth) were 72.4 ± 3.3 µm (70.5 - 79.9 µm) × 39.9 ± 2.5 µm (37.6 - 42.3 µm). The distinguishing morphological characteristics that have been noticed in ascarid worms were creamy white, cylindrical worm with triradiate lips with wide cephalic alae extending on both the lateral sides and filariform esophagus. In males, spicules were almost equal with the presence of precloacal chitinous-rimmed sucker. The prevalence of A. columbae infection was (63.1%) with a higher incidence in females (79.2%) than males (46.1%). The highest seasonal prevalence was observed in winter (92.5%), followed by summer and spring (87.5% and 55%), respectively while, the lowest prevalence was observed in autumn (17.5%). The intensity of worms in the infected intestine varied from 5 to 120 adult worms. The histopathological examination revealed the presence of chronic diffuse moderate catarrhal enteritis with roundworms in the lumen. Infected birds showed a significant increase in nitric oxide and MDA levels while serum zinc levels were lowered in infected pigeons. Infected pigeons revealed a marked increase in IL1-β and TNFα than apparently healthy ones., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The efficacy of commercially available anthelmintics against mature and immature stages (including ovicidal effects) of two Australian field isolates of Ascaridia galli was evaluated in two separate experiments. The anthelmintics tested were levamisole (LEV), piperazine (PIP) and flubendazole (FBZ) plus LEV-PIP. A total of 192 artificially trickle-infected young cockerels (96 birds per isolate) were randomized into sixteen experimental groups of 12 cockerels each (7 treatments and 1 untreated control per isolate). Chickens received label-recommended doses of LEV (28 mg/kg), PIP (100 mg/kg) or LEV-PIP co-administered at their full individual doses as a single oral dose or in group drinking water at recommended concentrations of 0.8 mg/ml or 2.5 mg/ml over eight hours for 1 and 2 days respectively and FLBZ (30 ppm) in the feed over 7 days. Anthelmintic efficacies were assessed by worm count reduction (WCR%) and excreta egg count reduction (EECR%) estimated by two methods. Ten days post treatment, all untreated control birds harboured mixed worm population of 10.1 and 12.3/bird for each isolate respectively which was significantly higher (P < 0.0001) than counts in all treatment groups. Luminal or histotrophic larvae comprised 50-57 % of the total worm count. For LEV, PIP and LEV-PIP, individual oral administration provided a somewhat higher efficacy than group medication in drinking water. EECR% values were inconsistent with WCR% and found to be only an indicator of efficacy against adult worms. All developmental stages of the two A. galli isolates were highly susceptible to FLBZ (100 %) followed by LEV-PIP (92.4-100 %) and LEV (87.7-100 %). PIP exhibited good efficacy against adult worms (92-97 %) but reduced efficacy against luminal (79-84 %) and histotrophic (61-72 %) larvae. Embryonation capacity of eggs recovered from worms expelled after treatment with LEV (47-54 %), PIP (44-54 %) or LEV-PIP (45-48 %) did not differ from those from untreated birds (50-51 %) whereas eggs from FLBZ treated worms had a significantly lower (P < 0.05) capacity to embryonate (≤ 2 %). Put together, our results demonstrate no evidence of resistance of the test A. galli isolates to the tested anthelmintics but a significant advantage of FLBZ, followed by LEV-PIP and LEV over PIP in the control of A. galli, specifically with regard to immature stages. A. galli worms expelled after treatment with LEV, PIP or their combination, but not FLBZ contain viable eggs. This has epidemiological implications and may also provide an option for isolating eggs from mature worms for A. galli propagation experiments without having to sacrifice birds., (Copyright © 2021 Elsevier B.V. All rights reserved.)

With the continued growth of free-range egg production, the importance of the chicken roundworm Ascaridia galli is increasing. Investigations into this parasite would be facilitated by the availability of characterised strains and clear guidelines on optimal methods of multiplication and maintenance. Currently, there is lack of well-defined in vivo models for maintaining A. galli and the potential of using host immunosuppression to boost parasite development and worm egg output has not been investigated. To determine the most efficient way of propagating A. galli in young chickens an experiment with a 2 × 3 × 4 × 2 factorial design involving age of chicken at infection (day-old or 14 days old), immunosuppression (dexamethasone (DEX), cyclophosphamide (CY) or sham), infective egg dose (0, 100, 300 or 900 embryonated eggs/bird) and time of worm recovery after infection (8 or 10 weeks post-infection) was conducted. The experiment used a total of 384 layer cockerel chicks. Infection was delivered orally in 3 split doses over one week and immunosuppressants were administered by intramuscular injection concurrently with the infections. Body weight, excreta egg counts, intestinal worm count and worm establishment rate were assessed. The only sign of ascaridiosis noted was mild diarrhoea at the time of slaughter in some birds with a significant- positive association with worm count. Infection caused a significant dose dependent reduction in body weight in non-immunosuppressed birds but this effect was ameliorated by immunosuppression. Age at infection had no significant effect on the studied variables although both worm and egg counts were numerically higher in the day-old infected groups. Egg dose significantly influenced the prevalence of infection, worm establishment rate, worm egg production and mean worm count. The 300 and 900 egg doses resulted in significantly higher worm count and egg production than the 100 egg dose. A significant negative correlation was observed between egg dose and worm establishment rate indicating an inverse relationship. Immunosuppression with DEX, but not CY resulted in significantly higher mean worm burden than in control chickens with excreta egg counts also considerably higher in DEX treated birds. Our results suggest that trickle infection at day-old with infective doses of 300 eggs coupled with immunosuppression with DEX would provide the most efficient way to propagate A. galli worms in vivo, as using older birds or a higher egg dose did not provide any advantage., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Since the EU ban on battery cages, many studies have listed Ascaridia galli and Heterakis gallinarum as the most common roundworms in the European laying hen population. A complicating factor is that the eggs of these parasites are almost identical. Thus, lack of molecular diagnostic approaches has driven epidemiological studies to take on necropsy for species discrimination, which is labor and cost intensive. Here, we describe a novel diagnostic tool based on droplet digital PCR for simultaneous identification and absolute quantification of the eggs of both of these ascarids in chickens' droppings using two different genus-specific primer-probe sets targeting the second internal transcribed spacer region (ITS-2) in the nuclear ribosomal (rRNA) gene array. No cross-reaction was observed when different combinations of DNA and species-specific primers and probes were tested. The lowest obtained frequency threshold for the detection of H. gallinarum in the presence of a constant A. galli DNA concentration was determined to be 0.8 %. After validation, we used the assay to analyze field samples collected from several Swedish laying hen farms. Out of 134 samples, 86 (64 %) were positive for A. galli while 11 (8.3 %) samples were positive for H. gallinarum. These samples were initially analyzed with flotation technique for detection of ascarid eggs. The results of the Cohen's kappa indicated substantial agreement (85.8 %) between the two tests. In conclusion, we have validated a novel molecular-based diagnostic tool for quantification and differentiation between intestinal parasites of major importance in chickens with high precision. Although this study focuses on identification of parasites of laying hens, the findings may well have a bearing on all types of chicken production systems. The present study lays the groundwork for future research into epidemiology of these two important chicken parasite species., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

We analyzed the nuclear ribosomal internal transcribed spacer (ITS) 1 and ITS2 sequences for Bangladesh isolates of Ascaridia galli, and we determined that the sequences were unreliable as molecular markers for distinguishing A. galli from other Ascaridia species, because the sequences showed high identity with that of A. columbae. However, the ITS1 sequences were available for designing PCR primers distinguishable between Ascaridia galli and Heterakis spp. Bangladesh isolates of A. galli constituted a monophyletic clade along with other geographical isolates in the cytochrome c oxidase subunit I (COI) phylogenetic tree, however, we could not clarify the phylogenetic relationships between A. galli and other Ascaridia spp., because their available sequences in GenBank were very few. The developed PCR method using DNA from A. galli and Heterakis spp. eggs would enable differential diagnosis of the individual infections in the future.

Eggs oviposited by Ascaridia galli females in artificial media are commonly used as a source of infective material. We investigated the rate of egg production by cultured mature females (n = 223), and changes in egg viability under different storage and incubation conditions. Eggs recovered after 1, 2 or 3 days of culture were subjected to either (1) storage in water at 4°C (1, 4 or 8 weeks) followed by incubation in 0.1 N H2SO4 at 26°C (2, 4 or 6 weeks); or (2) prolonged storage at 4°C (up to 14 weeks). Egg development and viability was assessed by morphology coupled with a viability dye exclusion test of hatched larvae. Of the 6,044 eggs recovered per mature female 49.2, 38.5 and 12.3% were recovered on days 1, 2 and 3 of worm incubation respectively with similar initial viability (≥99%) between days. Eggs recovered on different days had only minor differences in viability after storage. The prolonged storage period at 4°C significantly affected both viability and embryonation ability resulting in decline in viability of 5.7-6.2% per week. A smaller but significant decline in egg (2.0%) and hatched larval (1.4%) viability per week of incubation at 26°C was also observed. We conclude that storage and incubation conditions, not the day of egg recovery, are the main factors affecting A. galli egg viability. Our findings indicate that under aerobic conditions storage at 26°C may be preferable to 4°C whereas other studies indicate that under anaerobic conditions storage at 4°C is preferable.

Worm control is an important aspect of the successful management of the egg production industry. Of particular concern is Ascaridia galli, which at high parasite loads affect health and production in layers. Application of a targeted treatment strategy (TT) to control A. galli has shown promise. The purpose of this study was to explore the effect of such a strategy on welfare indicators and production performance of layers. Six flocks (F1-6) on a commercial farm were allocated to three treatment groups. Flocks F1 and F4 were treated (TT) with fenbendazole at 22, 27 and 36 weeks post-placement (WPP). Flocks F2 and F5 were treated at 27 WPP (conventional treatment, CT) and hens in flocks F3 and F6 served as untreated (UT) control groups. At 19, 35 and 45 WPP twenty-five hens plus thirty eggs per flock were randomly selected. Hens were weighed and their plumage conditions (PC) were assessed. The eggs were subjected to various external and interior quality analyses. Production data such as number of eggs/hen/week, egg mass and feed conversion ratio (FCR) were calculated from raw data obtained from all flocks on a weekly basis. The number of eggs/hen/week, egg mass and FCR were higher (P < 0.05) in the TT flocks and hens had better PC both at 35 and 45 WPP compared with other flocks. No differences in body weight and physical egg quality were observed between groups except for egg shell strength which was higher (P < 0.05) in the CT flocks. These data suggest that better production performance and plumage, which suggests improved health, can be achieved through the application of a TT strategy. The insights gained from this research should help to justify the extra cost and labor associated with the TT strategy., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This study evaluated the germination capacity of Pochonia chlamydosporia (VC4) fungus after its passage through the gastrointestinal tract of domestic chickens and its interaction with Ascaridia galli and Heterakis gallinarum eggs. Twenty-two domestic chickens were divided in two groups: control group (G1) received shredded corn substrate without VC4; and treatment group (G2) received a single dose of 29 g corn substrate containing 3.3 × 10 6 conidia/chlamydospores (VC4). Subsequently, chicken fecal samples were collected at intervals of 0, 6, 8, 10, 12, 18 and 24 h. Petri dishes from fecal samples of the treated group (G2) were subdivided (G2a and G2b), and then replicated in 2% agar-water medium for the microbiological test. After VC4 growth, approximately 200 eggs of A. galli (G2a) and H. gallinarum (G2b) were added to each subgroup to evaluation of ovicidal activity. There was fungal viability after passage through chicken gastrointestinal tract and egg predation of 59.9% and 43.2% for A. galli and H. gallinarum, respectively. The present work demonstrates the ability of the fungus P. chlamydosporia to survive after passing through the gastrointestinal tract of domestic chickens, an extreme environment (low pH, enzymes, microbiota and mechanical action), and still germinate after being excreted with feces., (Copyright © 2020 Elsevier Inc. All rights reserved.)

This study provides insights about the diversity, prevalence and distribution of alpine wild galliformes gastrointestinal parasite community, trying to fill a gap in the scientific information currently available in scientific literature. The analysis included three host species: 77 rock partridge (Alectoris graeca saxatilis), 83 black grouse (Tetrao tetrix tetrix) and 26 rock ptarmigan (Lagopus muta helveticus) shot during the hunting seasons 2008-2015. Parasites isolated were Ascaridia compar, Capillaria caudinflata and cestodes. The rock ptarmigan was free from gastrointestinal parasites, whereas the most prevalent helminth (37%) was A. compar in both black grouse and rock partridge. C. caudinflata occurrence was significantly higher in black grouse (prevalence = 10%, mean abundance = 0.6 parasites/sampled animal) than in rock partridge (prevalence = 1.20%, mean abundance = 0.01 parasites/sampled animal). Significant differences were detected among hunting districts. A. compar was found with a significant higher degree of infestation in the hunting districts in the northern part of the study area whereas cestodes abundance was higher in Lanzo Valley. Quantitative analysis of risk factors was carried out using a generalized linear model (GLM) only on the most common parasite (A. compar). Latitude was the only factors associated with infestation risk (OR = 52.4). This study provides information on the composition and variability of the parasite community in the alpine Galliformes species.

Background: Parasitism is a complex problem that is often ignored in companion animals, including birds, unless it develops into a severe clinical disorder. The present study was, therefore, aimed to investigate the presence of the gastrointestinal nematode infecting the domestic pigeon and provide a complete morphological description and clarify its taxonomic position through phylogenetic analysis of the ITS1-5.8s-ITS2 rDNA gene region., Materials and Methods: During the current study, a total of twenty-six domestic pigeons, Columba livia domestica, were collected and internal organs examined for helminth detection. Using light and scanning electron microscopy, the recovered parasite species are studied. In addition, the selected gene region was obtained and sequenced using appropriate primers that aid in the formation of the phylogenetic dendrogram for the recovered parasite species with others retrieved from GenBank., Results: Morphological examination showed that this nematode parasite belongs to the Ascaridiidae family within the genus Ascaridia. The material was assigned to the previously described Ascaridia columbae by providing all the characteristic features as the presence of a mouth opening surrounded by three tri-lobed lips; each lip has two triangular teeth with a spoon-like structure, cephalic papillae and amphidal pores on lips surface, presence of lateral cuticular alae and pre-cloacal sucker, 10 pairs of caudal papillae, and two equal spicules in male worms. The morphological investigations of this species were supplemented by molecular analysis of ITS1-5.8s-ITS2 rDNA gene region. The data showed that the present A. coulmbae is deeply embedded in the Ascaridia genus with a 74-99% sequence similarity to other species in the Chromadorea class. Ascaridiidae appears as monophyly and represented as a sister group to Heterakidae. The ascaridiid species examined belong to the Ascaridia genus and displaced a close relationship with the previously described A. coulmbae (gb| KF147909.1, gb| AJ001509.1, gb| KC905082.1, gb| JQ995321.1, gb| JX624729.1) as putative sister taxa., Conclusion: The present study revealed that the species Ascaridia is the first account of this genus as an endoparasite from the domestic pigeon inhabiting Saudi Arabia. Therefore, the combination of morphological and molecular studies helps to identify this species correctly and identified as Ascaridia columbae.

Competing Interests: Declaration of Competing Interest Authors disclose all relationships or conflict of interests that could have direct or potential influence or impart bias on the work.

Gastrointestinal nematodes are re-emerging in countries where the popularity of free-range poultry production systems is increasing. Amongst all gastrointestinal nematodes, Ascaridia galli is of significant concern due to the parasite's direct life cycle and ability to survive extreme environmental conditions. In laying hens, A. galli parasites have been associated with reduced health, welfare, immunity, and egg production. Direct losses are caused by obstruction and damage of the intestinal tract in hens when high worm burdens are present. These result in reduction in egg production and body weight of infected laying hens, consequently leading to significant economic losses for farmers. Furthermore, heavy infections with A. galli may lead to increased mortality within the flock. Indirect losses are due to suppression of immune system function which can increase susceptibility to secondary infections. Infection with A. galli can also alter nutrient utilization and absorption. Levels of anti- A. galli serum and egg yolk antibodies increase following A. galli infection. Elevated antibodies can be used as an indicator of current or previous infections and therefore can be used as a diagnostic tool. The impact of A. galli on hen health and welfare manifests through the depletion of liver lipid reserves and increased use of energy reserves to mount immune responses against the parasite. This review highlights the variable effects of A. galli infection on the performance, health, egg quality, and emphasizes especially on immune responses of free-range laying hens as well as it evaluates various potential detection methods and preventive and control measures of this parasitic disease., (© 2019 Poultry Science Association Inc.)

Purpose: Heterakidosis is a common parasitic infection caused in domestic birds by Heterakis species: Heterakis gallinarum, H. isolonche, and H. dispar. Among them, the best described species is H. gallinarum, noted mainly in gallinaceous birds. In waterfowl, H. dispar is the predominant species. The variations in morphology and host specificity qualify H. dispar as a different species, but the phylogenetic relationships between heterakids were unclear for a long time, because of a lack of H. dispar sequences., Methods: The authors provided the molecular data for H. dispar and analyzed the obtained sequences of the partial 18S rRNA gene and region ITS1-5.8SrRNA-ITS2 with the homological sequences., Results: The 18S rRNA PCR product of H. dispar was about 800 bp, and the ITS-5.8S-ITS2 PCR product was about 920 bp, noticeably smaller size compared to H. gallinarum product. The BLAST analysis of H. dispar 18S sequence showed a 99% similarity with the sequences of Heterakis gallinarum and Ascaridia galli, 98% with A. nymphii, but only 94% with the sequence of Heterakis sp. Our ITS sequence of H. dispar was almost identical to the H. isolonche isolate, there is only one nucleotide of difference among the 943 sites analyzed. It also showed a lower similarity to the ITS sequences of H. gallinarum (88%), H. spumosa (87%), and H. dahomensis (87%)., Conclusions: In our phylogenetic analysis, it is the first attempt at the reconstruction of relationships within this superfamily Heterakoidea based on 18S rDNA and ITS region.

An important factor in efficient production of poultry is management of parasites. Ascaridia dissimilis is the most prevalent small intestinal nematode parasite of turkeys with up to 100% of flocks infected. High worm burdens can cause necrotic enteritis leading to high mortality in flocks. Recently, we were made aware of multiple cases where high burdens were seen at slaughter despite the administration of anthelmintics at frequent intervals, suggesting that resistance may have evolved in A. dissimilis. To address this issue, we obtained eggs of A. dissimilis from 4 commercial turkey farms and performed controlled efficacy tests to determine if fenbendazole resistance was present. Three farms had histories of frequent use of fenbendazole and worms found at slaughter, suggesting they may have resistance, and one was an organic farm where we had no additional history other than the farm had transitioned to organic production a few years earlier. For each worm isolate there were 2 treated and 2 untreated groups containing 9 birds each, with all groups being replicated in 2 separate rooms. Birds were infected with approximately 200 infective eggs, and treated groups received fenbendazole in the water (SafeGuard Aquasol, 1 mg/kg) for 5 consecutive days starting on day 24 post-infection. One week after the last treatment birds were necropsied, intestinal contents were collected and worms enumerated. Three of the four isolates demonstrated greater than 99% efficacy, indicating they were fully susceptible to fenbendazole. However, the fourth isolate demonstrated a significantly reduced efficacy of 63.89%, indicating the presence of resistance. Interestingly, this was the organic farm, whereas the 3 farms with "suspected" resistance all turned out to be fully susceptible. Given that 1 randomly acquired isolate of A. dissimilis, out of 4 tested, demonstrated resistance in this study, fenbendazole resistance may be a much larger problem on turkey farms than is currently recognized. Additional studies are needed to determine the prevalence of resistance, as well as the economic impact that resistant A. dissimilis have on turkey production., (© 2019 Poultry Science Association Inc.)

The detection and quantification of nematode eggs using fecal egg count techniques have an irreplaceable role in equine parasitic control. The reliability, particularly precision and accuracy, of individual techniques have been described only for strongylid infections. The aim of this study was to compare three fecal egg count techniques used for the detection of the two most common equine nematode infections: strongylid and ascarid. The Simple McMaster, Concentration McMaster and Mini-FLOTAC techniques were tested on spiked fecal samples with various levels of egg concentration (50, 100, 200, 500, 1000 and 3000 eggs per gram) and naturally infected mixed strongylid-ascarid samples with 30 replicates. The Simple McMaster, Concentration McMaster and Mini-FLOTAC techniques had precision coefficients of variation of 44.33, 35.64 and 18.25% for the strongylid infection and 62.95, 35.71 and 18.95% for the ascarid infection, and percent accuracies (mean count/number of eggs spiked) of 97.53, 88.39 and 74.18% for the strongylid infection and 65.53, 83.18 and 90.28% for the ascarid infection, respectively. Accuracy depended greatly on the type of nematode, but precision did not. The Mini-FLOTAC technique was more precise than the Simple and Concentration McMaster techniques regardless of nematode type. Simple McMaster was the most accurate technique for detecting strongylid eggs, and Mini-FLOTAC was the most accurate technique for detecting ascarid eggs. Our results indicated that none of the current techniques were universally and sufficiently reliable for the simultaneous quantification of both of these common equine nematodes., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Coccidiosis is one of the most economically important diseases of the poultry, around the globe. In order to assess seasonal and age-wise prevalence of coccidiosis individually and with concurrent infections, this study was conducted on commercial poultry farms in and around Multan division, province of Punjab, Pakistan. A total of 28,126 boilers, 4,052 layers and 7,699 golden bird samples, provided by regional farmers and consultants, were examined by microscopy for the diagnosis of coccidiosis. Based on postmortem lesions, several infections including coccidiosis, viral, and miscellaneous diseases were catalogued, whereas samples were cultured to identify concurrent bacterial diseases. Cumulative analysis of this large set of samples revealed a prevalence of 14.16, 11.01, and 19.57% in broiler, layer and golden birds, respectively. Ascaridia galli (A. galli) showed a higher prevalence in layer (2.47%) compared to golden (0.21%) birds (P < 0.01). Amongst all concurrent infections investigated, bacterial infections were identified in highest percentage of samples (59.24%; P < 0.05). The age-wise and season-wise prevalence of coccidiosis and A. galli was found to be significant (P < 0.05). During the study period, an estimated of 10.69 (coccidiosis) and 0.29 (A. galli) million poultry birds were treated or provided prophylaxis and supportive therapy at Pakistani commercial poultry farms. Depending upon the prevailing market conditions, the total economic losses (treatment, prophylaxis, and supportive therapy) from coccidiosis and A. galli were estimated to be US $45,405.00 and 2,638.50, respectively, while production (weight and eggs) losses for broiler (US$ 2,750,779.00), layer and golden, young (US$ 13,974.98 and 50,228.76) and adult (US$ 104.74 and 203.77) were estimated. Unit loss of coccidiosis with concurrent diseases and intestinal worm was estimated to be US$ 0.005 and 0.01, respectively. These results highlight the potential impact of coccidiosis individually and with concurrent infections on the poultry productivity and will inform farmers, policy makers, and other governmental and non-governmental stakeholders on the use of control and management measures in containing these infections., (© 2018 Poultry Science Association Inc.)

Ascaridia galli is one of the most abundant nematode parasites in poultry. A. galli infections can significantly impact the profitability of egg farms and have negative implications for bird health and welfare. The main objectives of this study were to determine whether A. galli specific antibodies in egg yolks can be used to detect prior or current exposure to A. galli in laying hens, and to distinguish between eggs obtained from caged and free-range hens. Twenty-two laying hen flocks from different production systems (10 free-range, 2 barn-housed, and 9 caged flocks) were enrolled in the study. An in-house enzyme-linked immunosorbent assay was used to analyze levels of A. galli specific antibodies in yolk. The numbers of A. galli eggs in hen excreta were also determined in a subset of farms. Free-range flocks had higher and also more variable levels of anti-A. galli antibodies in the egg yolk compared to those of the cage flocks (0.50 ± 0.39 vs. 0.16 ± 0.13 OD units) (P < 0.001). Results also confirmed that excreta from free-range and barn-housed flocks contained higher numbers of A. galli eggs than did excreta from caged flocks in which no A. galli eggs were detected. In conclusion, analysis of anti-A. galli antibodies in the egg yolk can be used to detect worm exposure in commercial layer flocks. However, the method used in this study cannot be used in isolation to distinguish between eggs from cage and free-range production systems as anti-A galli antibodies were detected in egg yolk samples from all production systems, and the range of antibody levels overlapped between production systems.

Worm expulsion is known to occur in mammalian hosts exposed to mono-species helminth infections, whilst this phenomenon is poorly described in avian hosts. Mono-species infections, however, are rather rare under natural circumstances. Therefore, we quantified the extent and duration of worm expulsion by chickens experimentally infected with both Ascaridia galli and Heterakis gallinarum, and investigated the accompanying humoral and cell-mediated host immune responses in association with population dynamics of the worms. Results demonstrated the strong co-expulsion of the two ascarid species in three phases. The expulsion patterns were characterized by non-linear alterations separated by species-specific time thresholds. Ascaridia galli burden decreased at a daily expulsion rate (e) of 4.3 worms up to a threshold of 30.5 days p.i., followed by a much lower second expulsion rate (e = 0.46), which resulted in almost, but not entirely, complete expulsion. Heterakis gallinarum was able to induce reinfection within the experimental period (9 weeks). First generation H. gallinarum worms were expelled at a daily rate of e = 0.8 worms until 36.4 days p.i., and thereafter almost no expulsion occurred. Data on both humoral and tissue-specific cellular immune responses collectively indicated that antibody production in chickens with multispecies ascarid infections is triggered by Th2 polarisation. Local Th2 immune responses and mucin-regulating genes are associated with the regulation of worm expulsion. In conclusion, the chicken host is able to eliminate the vast majority of both A. galli and H. gallinarum in three distinct phases. Worm expulsion was strongly associated with the developmental stages of the worms, where the elimination of juvenile stages was specifically targeted. A very small percentage of worms was nevertheless able to survive, reach maturity and induce reinfection if given sufficient time to complete their life cycle. Both humoral and local immune responses were associated with worm expulsion., (Copyright © 2018 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Ascaridia (A.) galli is one of the most commonly occurring nematodes in poultry worldwide, often in hens and broiler chickens. The infection with Ascaridia galli in free-range chickens was even 70%. There is not much information about A. galli genetic features. The present study was conducted to assess the genetic diversity of A. galli isolated from hens in Poland by analyzing the nucleotide sequence of the region ITS1-5.8rRNA-ITS2 and to define its homology within the family Ascaridiidae. Adult A. galli were collected from the intestines of naturally infected hens from two flocks of free-run laying hens from the Wielkopolska region in Poland. From all parasites an identical ITS1-5.8rRNA-ITS2 sequence was obtained, which was homologous in 99% with A. columbae (JQ995321.1) sequence. The high homology sequences of A. galli (KX683286) from Poland and A. columbae (JQ995321.1) isolate from the USA, support the observations of other authors suggesting that A. galli and A. columbae might be closely related. It is the first whole ITS1-5.8rRNA-ITS2 of A. galli in the GenBank database, so there is not enough data for detailed phylogenetic analysis of A. galli. Detailed genetic analysis is necessary to get better insight into the birds' Ascaridia species.

Present study was performed to identify the species of ascarids from macaw parrot, Ara chloroptera, in China. Total 6 ascarids (3 males and 3 females) were collected in the feces of 3 macaws at Guangzhou Zoo in Guangdong Province, China. Their morphological characteristics with dimensions were observed under a light microscope, and their genetic characters were analyzed with the partial 18S rDNA, ITS rDNA and nad4 gene sequences, respectively. Results showed that all worms have no interlabia but male worms have two alate spicules, well-developed precloacal sucker and a tail with ventrolateral caudal alae and 11 pairs of papillae. The partial 18S rDNA, ITS rDNA and nad4 sequences were 831bp, 1015bp and 394bp in length, respectively. They showed the highest similarity of 99.8% (18S rDNA) with Ascaridia nymphii, 93.8% identities (ITS rDNA) with A. columbae and 98.5% to 99.5% identities (nad4) with Ascaridia sp. from infected parrot. All Ascaridia nematodes from the macaws were clustered into one clade and formed monophyletic group of Ascaridia with A. columbae and A. galli in two phylogenetic trees. It is observed that the combining morphological and sequencing data from three loci, the present Ascaridia species was identified as Ascaridia nymphii, which is the first record of A. nymphii from macaw parrot in China., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A study was conducted to determine the performance, egg quality, and liver lipid reserves of laying hens exposed to ranges contaminated with Ascaridia galli. Sixteen-week-old Lohmann Brown laying hens (n = 200) were divided into 4 treatments with 5 replicates containing 10 hens per pen. Hens of treatment 1 [negative control (NC)] ranged on a decontaminated area, and hens of treatments 2 (low infection) and 3 (medium infection) ranged on areas previously contaminated by hens artificially infected with 250 and 1,000 embryonated A. galli eggs, respectively. The hens of treatment 4 [positive control (PC)] ranged on areas previously contaminated by hens artificially infected with 2,500 embryonated A. galli eggs, and in addition these hens were orally inoculated with 1,000 embryonated eggs. Results indicated that hens of the medium infection group had a higher number of intestinal A. galli worms and A. galli eggs in the coprodeum excreta (43.9 ± 4.0 and 3,437 ± 459 eggs/g) compared to hens of the low infection group (23.8 ± 4.0 and 1,820 ± 450 eggs/g) (P < 0.01) and similar worm counts to PC hens (34.4 ± 4.0 and 2,918 ± 474) (P > 0.05). Egg production, egg mass, feed intake, and feed conversion ratio (FCR) were not affected by A. galli infection (P > 0.05). Egg quality parameters (egg weight, shell reflectivity, shell weight, shell thickness, shell percentage, shell breaking strength, deformation, albumen height, Haugh unit, and yolk score) were not affected by A. galli infection (P > 0.05). Highly infected hens had lower liver lipid content (2.72 ± 0.51 g) compared to uninfected hens (4.46 ± 0.58 g, P < 0.01). The results indicate that exposure to ranges contaminated with A. galli resulted in infection of the ranging hens, but this did not affect egg production or egg quality. Infection with A. galli lowered the liver lipid reserves of the host significantly, suggesting infected hens use more energy reserves for maintenance and production.

Reliable methods for detection of A. galli infection using excreta egg count (EEC) and ELISA assays to determine A. galli specific IgY levels in serum and yolk samples were compared from hens infected naturally and artificially. Artificially infected hens were used to generate samples for analysis of preferred detection methods and to generate contaminated ranges for use in the naturally acquired infection study in which Lohmann Brown hens (n = 200) at 16 weeks of age were randomly assigned to four treatments with five replicate pens. Hens of negative control (NC) ranged on a decontaminated area, hens of low infection, medium infection and positive control (PC) ranged on the areas previously contaminated by hens artificially infected with 250, 1000 and 2500 A. galli eggs/hen, respectively. Additionally, hens of PC were orally infected with 1000 A. galli eggs/hen. Anti A. galli antibody levels in hen serum (SIgY) and yolk (YIgY) were measured before range access, and 2, 7 and 12 weeks after access to the contaminated ranges. In a natural infection study, eggs were detected in the excreta of all hens 4 weeks after range access, with the exception of NC in which no eggs were detected. EEC increased to reach maximum value (2204 ± 307 eggs/g) after 11 weeks of range access and then declined at 12 weeks (905 ± 307eggs/g) (p < 0.01). While SIgY OD values were not different in hens between any groups before range access, after 2 weeks, both SIgY and YIgY gradually increased in hens of PC (1.17 ± 0.03 and 0.88 ± 0.04) and medium infection (1.07 ± 0.03 and 0.96 ± 0.04) compared to low infection (0.38 ± 0.03 and 0.29 ± 0.04) (p < 0.01) and NC. After 12 weeks, SIgY were similar in hens of PC, medium and low groups whereas YIgY was higher in hens of low infection group (p < 0.01). Sensitivity of the serum and egg yolk antibody levels assay to detect A. galli infection was 100% and 96%, respectively, whereas the pooled EEC method yielded a sensitivity of 93%. The results of this study suggest that hens naturally infected with A. galli produce both SIgY and YIgY at different levels depending on the infection intensity and duration of exposure which allows the diagnosis of prior infection or early diagnosis of current infection. Use of the practical and non-invasive method of yolk sample analysis for detecting IgY can be just as informative as using serum samples to detect A. galli infection., (Crown Copyright © 2018. Published by Elsevier B.V. All rights reserved.)

Background: The nematophagous fungus Pochonia chlamydosporia can degrade ascarid (e.g. Ascaridia galli) eggs in agar and soil in vitro. However, it has not been investigated how this translates to reduced infection levels in naturally exposed chickens. We thus tested the infectivity of soil artificially contaminated with A. galli (and a few Heterakis gallinarum) eggs and treated with P. chlamydosporia. Sterilised and non-sterilised soils were used to examine any influence of natural soil biota., Methods: Unembryonated eggs were mixed with sterilised (S)/non-sterilised (N) soil, either treated with the fungus (F) or left as untreated controls (C) and incubated (22 °C, 35 days) to allow eggs to embryonate and fungus to grow. Egg number in soil was estimated on days 0 and 35 post-incubation. Hens were exposed to the soil (SC/SF/NC/NF) four times over 12 days by mixing soil into the feed. On day 42 post-first-exposure (p.f.e.), the hens were euthanized and parasites were recovered. Serum A. galli IgY level and ascarid eggs per gram of faeces (EPG) were examined on days -1 and 36 (IgY) or 40 p.f.e. (EPG)., Results: Egg recovery in SF soil was substantially lower than in SC soil, but recovery was not significantly different between NF and NC soils. SF hens had a mean worm count of 76 whereas the other groups had means of 355-453. Early mature/mature A. galli were recovered from SF hens whereas hens in the other groups harboured mainly immature A. galli. Heterakis gallinarum counts were low overall, especially in SF. The SF post-exposure IgY response was significantly lower while EPG was significantly higher compared to the other groups., Conclusions: Pochonia chlamydosporia was very effective in reducing ascarid egg numbers in sterilised soil and thus worm burdens in the exposed hens. However, reduced exposure of hens shifted A. galli populations toward a higher proportion of mature worms and resulted in a higher faecal egg excretion within the study period. This highlights a fundamental problem in ascarid control: if not all eggs in the farm environment are inactivated, the resulting low level infections may result in higher contamination levels with associated negative long-term consequences.

Factors affecting the development of Ascaridia galli-specific humoral responses and their protective roles are largely unknown. We investigated the effects of time and infection dose on A. galli-specific IgY antibody levels following experimental infection. The acquisition and development of new infections and reinfections were also monitored by using tracer birds. Relationships between the retrospective IgY and the final worm burden of the birds were investigated to determine whether humoral immune responses generated during infection provide protection to the host animal. Young chickens were infected (+) with either 100 or 1000 embryonated eggs of A. galli (100+: n = 45; 1000+: n = 45) or kept as uninfected controls (CON: n = 10). Uninfected birds were also added to each infection group as tracer (T) birds (T100+; n = 5 and T1000+; n = 5). Faecal egg counts and IgY antibody concentrations in plasma and egg yolk were determined at selected intervals. Final worm burdens were quantified at 28 weeks post infection (wpi). The plasma antibody (PAB) and egg yolk antibody (EAB) levels of experimentally infected birds were compared to those of control and tracer birds throughout the study period, and PAB levels were found to depend initially on the infection dose but thereafter mainly on reinfections. Starting at wpi 2, 1000+ had consistently higher PAB levels than CON did (P < 0.05). With exceptions at wpi 0, 2 and 12, PAB levels were also higher (P < 0.05) or tended to be higher (P < 0.10) in 100+ than in CON. An earlier and higher increase was observed in the PAB levels of T1000+ than in those of T100+, implying that (re-)infection occurrence depended on the infection dose. Although 1000+ showed higher (P < 0.05) EAB levels than CON did at both wpi 14 and 18, EAB levels were higher in 100+ than in CON only at wpi 28 (P < 0.05). The total worm burdens in the initial experimentally infected birds were similar (P = 0.257); they were also highly comparable between experimentally and naturally infected birds, indicating that final worm burden was mainly determined by the naturally occurring infections resulting from continuous exposure. When all available information on the retrospective plasma and egg yolk IgY levels was collectively evaluated to estimate the larval or total worm burdens of the experimentally infected birds, both PAB and EAB levels at particular wpi were significantly associated with worm burden, especially with larval count. In conclusion, our data support the hypothesis that the number of larvae, rather than the number of mature worms, affects the antibody levels in both plasma and egg yolk. Despite the increased levels of A. galli-specific antibodies in plasma and egg yolk throughout the study period, only a weak indication was found that antibodies might be directly associated with protection., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study was conducted to determine the effect of Ascaridia galli infection on free-range laying hens. Lohmann Brown laying hens (n = 200) at 17 wk of age were allocated to 4 treatment groups (n = 50 per group), each with 5 replicate pens of 10 hens. Hens in 3 treatment groups were orally inoculated with different doses of embryonated A. galli eggs: low (250 eggs), medium (1,000 eggs), and high (2,500 eggs) levels, whereas hens of the control group were not infected. Infection levels were monitored using excreta egg counts and mature A. galli worm counts in the intestine. Anti A. galli antibody titers (IgY) in the serum were measured prior to infection, and at 6, 11, 15, and 20 wk post infection (PI) and in egg yolk at 11 and 20 wk PI. Parameters evaluated included feed intake, egg production, egg weight, egg mass, FCR, liver weight, liver fat, and intra epithelial immune cell infiltration. The results showed no difference in feed intake, body weight, or FCR among any treatment groups (P > 0.05). Egg production was lower in the low infection group compared to other groups at 20 wk of age (P < 0.01). Serum IgY was higher in the infected groups' hens at 20 wk PI compared to control group hens (P < 0.01). Yolk IgY increased significantly over time and was higher in infected hens compared to hens of the control group at 11 and 20 wk PI (P < 0.001). No differences were observed in liver lipid content or intraepithelial lymphocytes infiltration among treatment groups. Ascaridia galli eggs in the coprodeum content and adult A. galli worm count were higher in infected hens compared to hens of the control group (P < 0.01). In conclusion, the effects of artificial infection with A. galli on the parameters investigated were minor, and egg yolk antibody may be a more reliable indicator of A. galli infection than serum antibody or excreta egg count., (© 2017 Poultry Science Association Inc.)

The present study was undertaken to find out the safety levels of fenbendazole in common peafowl. This bird, raised on aviaries and zoos, can be severely parasitized with Ascaridia galli (enteric worms) and Syngamus trachea (gapeworm) along with other parasitic worms. Fenbendazole is a highly effective benzimidazole-class anthelmintic in animals. The objective of this work was to provide target animal safety data in young peafowl and to demonstrate reproductive safety in adult birds. During the experimental study, diets containing fenbendazole at 0, 100, 200 and 300 ppm were fed for 21 days (three times the normal treatment duration). Data for feed consumption, feed conversion rate, and body weights were recorded for each bird in each group. Drug concentrations in different tissues of birds were determined to correlate concentrations with clinical observations, clinical pathology, and histologic findings. There were no morbidities or mortalities after study day 21. Additionally, there were no statistically significant treatment-related differences among above mentioned parameters. Analysis of fenbendazole concentrations in kidney, liver, leg/thigh, and breast muscle and skin with associated fat revealed that, even at the highest dose level used and with no feed withdrawal, fenbendazole concentrations were relatively low in these tissues. These findings indicate that fenbendazole has a relatively wide margin of safety in young peafowl and that the proposed dose of 100 ppm in the feed for 7 consecutive days is well within the margin of safety. In the reproductive safety study, five breeder peafowl farms fed fendbendazole at 100ppm for 7 days and collected data on hatching percentage of peahen eggs before and after treatment. Reproductive performance in peahen was not adversely affected.

Broilers commonly suffer from necrotic enteritis (NE). Other gastrointestinal infectious diseases affect poultry, including nematode infections which are considered a re-emerging disease in barn and free-range systems. The aim of this study was to characterize the immune response of broilers after artificial infection with NE and contrast these with responses to the nematode Ascaridia galli and determine whether immune parameters measured during the course of infection can be used to distinguish infected from uninfected birds. A total of 96 one-day-old male Ross 308 broiler chickens were used in this study. At 10 days of age, broilers were randomly assigned to one of the following treatment groups: control birds (n = 32), A. galli infected birds (n = 32), or NE infected birds (n = 32) and inoculated with the appropriate infective agents. The immune response of birds was monitored through evaluation of haematology parameters, acute phase protein production, and intraepithelial intestinal lymphocyte population changes at 11, 16, 20, and 32 days of age. T-helper cells (CD4 + CD8 - ) increased significantly over time, and were significantly higher in A. galli and NE compared to day 10 controls. In conclusion, α-1 glycoprotein levels can distinguish birds with NE from other birds, including those infected with A. galli; also T-helper cell numbers can distinguish both NE and A. galli from uninfected birds and thirdly, 10 days post infection is the best time point to evaluate the bird's immune response for A. galli infections.

The present study analyzed gastrointestinal helminth communities in 265 wild pigeons (Columba livia) living in the municipalities of São Paulo and Tatuí, state of São Paulo, Brazil, over a one-year period. The birds were caught next to grain storage warehouses and were necropsied. A total of 790 parasites comprising one nematode species and one cestode genus were recovered from 110 pigeons, thus yielding an overall prevalence of 41.5%, mean intensity of infection of 7.2 ± 1.6 (range 1-144) and discrepancy index of 0.855. Only 15 pigeons (5.7%) presented mixed infection. The helminths isolated from the birds were Ascaridia columbae (Ascaridiidae) and Raillietina sp. (Davaineidae). The birds' weights differed according to sex but this did not influence the intensity of infection. The overall prevalence and intensity of infection did not differ between the sexes, but the prevalence was higher among the birds from Tatuí (47.8%). The gastrointestinal helminth community of C. livia was characterized in the two areas studied and parasite homogeneity was observed over the 12 months analyzed at both locations. These results make contributions to the current literature on health aspects of wild C. livia populations.

Susceptability of Ascaridia galli to benzimidazole (BZ) was investigated using faecal egg count reduction test (FECRT), in ovo larval development test (LDT) and genetic markers (mutations at codons 167, 198 and 200 of β-tubulin gene). Six flocks (F1-F6) of a commercial laying hen farm with different number of exposure to BZ were recruited. The FECR was calculated by analyzing individual faeces (F1, F2, F4 and F5) before and 10 days after treatment. The LDT was performed on parasite eggs from pooled samples from F1 to F6 and LC50 and LC99 were calculated. DNA was extracted from 120 worms and sequenced for β-tubulin gene. In all flocks, the FECRs were above 95% (lower CI above 90%). No significant difference was observed (p > 0·05) among obtained LC50 (F1/F4 and F2/F5 vs F3/F6) in the LDT. However, LC50 and LC99 were higher than suggested values for declaration of resistance in other nematode species. No variation was observed in codon positions involved in BZ resistance. Overall, our results indicated lack of evidence of resistance to BZ in A. galli. More research is needed to confirm these results and to further optimize the existing tools for detection and monitoring of anthelmintic resistance in A. galli.

Eggs of chicken ascarids (Ascaridia galli and Heterakis spp.) are believed to be hardy and survive for long periods. However, this has not been evaluated quantitatively and our study therefore aimed to determine development and recovery of chicken ascarid eggs after burying in pasture soil. Unembryonated eggs were mixed with soil, placed in sealed nylon bags and buried at 7 cm depth in pasture plots April (spring, n = 72) and December 2014 (winter, n = 72). Eight randomly selected bags per season were used to estimate pre-burial egg recovery [0 week post-burial (wpb)]. Eight random bags were removed at 5, 12, 23, 38, 52, 71 wpb per season and additionally at 104 wpb for spring burial. The content of each bag was analysed for numbers and development stages of eggs. Eggs buried in spring were fully embryonated within 12 wpb. In contrast, eggs buried in winter were developing between 23 and 38 wpb, so that all viable eggs seemed to be fully developed by 38 wpb. About 90% eggs disappeared within 23 wpb (spring) and 38 wpb (winter). Small proportions (2-3%) of seemingly viable and infective eggs were still recovered up to 2 years after deposition. In conclusion, most eggs buried in temperate pasture soil seem to experience a heavy mortality within a few months after the deposition, especially during warm periods. However, a small proportion of eggs may survive and remain infective for at least 2 years.

Background: Classical faecal egg counts (FEC) provide less reliable diagnostic information for nematode infections in chickens. We developed an ELISA based on Ascaridia galli antigens and tested two hypotheses, as follows: (i) IgY antibodies developed against A. galli will also be useful to identify Heterakis gallinarum infections, and (ii) circulating antibodies stored in egg yolks are as good as plasma samples, so a non-invasive diagnosis is possible. The aim of this study, therefore, was to compare the diagnostic accuracy of the ELISA system with FEC, using both plasma and egg yolks from experimentally infected hens. In addition, naturally infected animals were evaluated to validate the assay., Results: The assay quantified large differences (P < 0.001) in plasma or in egg-yolk IgY concentrations between infected and uninfected animals in two experiments, each performed with either of the nematode species. The assay performed with high accuracy as quantified with the area under the ROC curve (AUC) values of > 0.90 for both nematodes using either plasma or egg yolks. Sensitivity of the assay was 94 and 93% with plasma and egg yolk samples, respectively, whereas FEC yielded in a sensitivity of 84% in A. galli experiment. Total test accuracy of the assay with plasma samples (AUC = 0.99) tended to be higher (P = 0.0630) than FEC (AUC = 0.92) for A. galli, while the assay with either sample matrix performed similar to FEC (AUC ≥ 0.91) for H. gallinarum. Among the three tests, the FECs correlated better with A. galli burden than the ELISA. Although 90% of naturally infected hens were correctly identified by the ELISA, 45% of the infected hens tested negative with FEC, indicating the validity of the higher test accuracy of the ELISA., Conclusions: Antigens of A. galli can be used successfully to identify H. gallinarum-infected animals, indicating that chickens develop cross-reactive antibodies against the two closely related species. Egg yolks are as informative as plasma samples, so that animal welfare-friendly sampling is possible. Although the assay with plasma samples reveals qualitative information of higher quality than FECs on the infection status of naturally infected birds, the latter is still a better tool to assess the intensity of A. galli but not of H. gallinarum infections.

Maternally derived antibodies can provide partial protection against certain bacterial and viral infections. We investigated whether chicks descending from nematode-infected hens are more resistant against Ascaridia galli, a prevalent gastrointestinal nematode, than chicks from nematode-free mothers. One-day-old chicks (N=153) from infected (mab+; maternal antibody+) or uninfected control dams (mab-) were experimentally infected with A. galli at two different levels (100 or 1000 eggs/chick). The worm burdens of the chicks were determined at 6 weeks post infection. There was a high correlation (r=0.89) between A. galli-specific antibody concentrations in dam plasma and egg yolk. There was no difference between worm burdens of chicks descending from infected or uninfected dams (P=0.892), indicating no maternally derived protection against A. galli. Chicks receiving the higher infection dose had higher worm burdens (P<0.05). Although there was no difference (P>0.05) between worm counts of female and male chicks infected with 100 eggs, females chicks infected with 100 eggs harboured longer and heavier female worms. We conclude that there is no protective maternal immunity against A. galli infection., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The efficacy of a sustainable deworming strategy based on targeted treatments (TT) against Ascaridia galli was investigated for the first time in laying hen flocks on a Swedish commercial farm. Three experimental protocols with different levels of treatment, e.g. targeted treatment (TT), conventional treatment (CT) and untreated (UT), were tested in randomly allocated flocks of two different bird hybrids. Every second week faecal egg counts (FECs) were determined from pooled faecal materials collected on trays (20×27cm) placed for a maximum of 12h on the litter belts. In the TT, anthelmintic administration (fenbendazole, 1mg/kg body weight for 5days) started at 22 weeks post placement (wpp) and was repeated twice when the pooled FECs surpassed the assigned threshold of 200 egg per gram faeces (EPG). The CT flocks were treated once at 27wpp using the same anthelmintic. Hens in the UT were not dewormed and served as controls. Additionally, FECs on cloacal contents, worm fecundity and worm burdens were determined at 19, 35 and 45wpp. None of the flocks became infected until after 16wpp. The cumulative pooled FECs at the end of the study were significantly (p<0.01) lower in the TT compared to both CT and UT. Although repeated treatment in the TT protocol did not affect the fecundity, a worm density-dependent increase in fecundity was observed. Cloacal FECs and the number of adult A. galli in TT at 35 and 45wpp were significantly lower compared to other flocks. This study provides evidence that the TT strategy is better in terms of lower worm burden and decreased cumulative environmental parasite egg numbers compared to CT strategy. The TT strategy should be considered as an alternative to the CT strategy with regard to A. galli control in commercial laying hens., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Infection with the poultry roundworm Ascaridia galli has increased in European countries due to the ban on battery cages. This study was conducted in two commercial laying hen flocks (F1 & F2) on different farms in central Sweden. The aims were to (1) investigate the efficacy of flubendazole (FLBZ, 1.43 mg/kg administered in drinking water for 7 days) against adult and larval stages including histotrophic larvae of A. galli, and (2) determine how long it took before the flocks were reinfected after deworming. Accordingly, 180 randomly selected hens were sacrificed before drug administration (bd), on day 3 and 7 during drug administration (dd), and on a weekly basis for up to five weeks post drug administration (pd). Intestinal contents and cloacal materials of each hen plus pooled faecal samples from manure belts were investigated to assess the worm burden and the parasite egg per gram faeces (epg). Additionally, drinking water, and serum and gastrointestinal digesta content samples obtained from ten treated animals were analyzed by HPLC to measure FLBZ and its reduced (R-FLBZ) and hydrolyzed (H-FLBZ) metabolites. No parasite eggs were observed in cloacal samples on day 21 and 28 pd on F1 and on day 21 pd on F2. The epg in manure decreased by 65% and 88% on day 3 dd and by 99% and 97% on day 35 pd on F1 and F2 respectively. Mean FLBZ concentrations quantified in duodenal contents ranged between 0.50 and 0.79 μg/g. Although, no histotrophic larvae were found dd, they reappeared one week pd (7 ± 7 F1, 0.5 ± 0.5 F2). Adult worms were found in both flocks before drug administration (44 ± 20 F1, 35 ± 25 F2), on day 3 dd (4 ± 3 F1, 2 ± 2 F2), and then not until day 35 (0.2 ± 0.6) on F1 and day 28 (0.4 ± 0.9) pd on F2. Thus, the only period in which no A. galli were found was on day 7 dd. Although FLBZ was highly efficient our results indicate that the birds were reinfected already within one week pd., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Ascaridia galli is one of the most common nematode affecting chickens. This study characterized A. galli parasites collected from South African village chickens of Limpopo (n=18) and KwaZulu-Natal (n=22) provinces using the 510bp sequences of cytochrome C oxidase subunit 1 gene of the mitochondrial DNA. Fourteen and 12 polymorphic sites were observed for Limpopo and KwaZulu-Natal sequences, respectively. Six haplotypes were observed in total. Haplotype diversity was high and ranged from 0.749 for Limpopo province to 0.758 for KwaZulu-Natal province isolates. There was no genetic differentiation between A. galli from Limpopo and KwaZulu-Natal provinces. The six South African haplotypes were unique compared to those published in the GeneBank sampled from Hy-line chickens raised under organic farming in Denmark. The utility of cytochrome C oxidase subunit 1 gene as a potential genetic marker for studying A. galli in village chicken populations is presented., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Experimental infection models for Ascaridia galli rely on the use of eggs isolated either directly from worm uteri or from host faeces. We investigated whether A. galli eggs isolated from the two sources differ in their embryonation ability. A. galli eggs originating from 12 worm infrapopulations were isolated both from faeces of the living host (faecal eggs) and directly from worm uteri after host necropsy (uterine eggs). The isolated eggs from each infrapopulation and source were incubated in Petri dishes (n=24) containing a potassium-dichromate (0.1%) medium for 28 days (d) at room temperature. Starting from the day of egg isolation (d0), in ovo larval development was evaluated every second day by examining morphological characteristics of 200 eggs/petri dish. A total of 72,000 eggs were classified into undeveloped, early development, vermiform or fully embryonated stages. Isolation procedures caused similar damage to uterine and faecal eggs (2.2% and 0.5%, respectively; P=0.180). The first sign of in ovo embryonic development in faecal eggs (7%) was observed during the 24-h period when faeces were collected. On d28, a higher percentage of uterine eggs remained undeveloped when compared with faecal eggs (58.6% vs 11.0%; P<0.001). Although a higher (P<0.001) percentage of faecal eggs entered both the early developmental and vermiform stages, which took place primarily within the first two weeks of incubation, there was no time-shift between the development of faecal and uterine eggs. Starting from day 10, higher (P<0.05) percentages of faecal eggs completed embryonation compared with uterine equivalents. Eggs from both sources reached a plateau of embryonation by the end of 2nd week of incubation, with faecal eggs having a greater than two-fold higher embryonation ability. Cumulative mortality was higher in uterine eggs (14.3%) than in faecal eggs (0.2%). We conclude that faecal eggs have a higher embryonation ability than uterine eggs possibly due to maturation differences., (Copyright © 2015 Elsevier B.V. All rights reserved.)