Genetic differences of Ascaridia galli egg output in laying hens following a single dose infection.

Groups of 20-week-old white (Lohmann LSL) and brown (Lohmann Brown) hens were reared under helminth-free conditions and both divided into two subgroups. One subgroup was artificially infected with 250 embryonated Ascaridia galli eggs, the other subgroup were kept as uninfected controls. During the following 12-month laying period individual faecal Ascaridia egg counts (FEC) were performed and bodyweight measured at monthly intervals. Laying performance and egg weights were determined daily. The mean FEC were significantly (P < 0.01) higher in infected white hens than in infected brown hens. The growth rate of the white hens was significantly (P < 0.05) higher in the control group, whereas in brown hens no significant group difference was observed. The laying performances and egg weights did not differ significantly (P > 0.05) between control and infected animals for either line. The estimated repeatabilities for mean log(10) FEC of different samples were reasonably high (0.16-0.54). Heritabilities for mean log(10) FEC were between 0.13 and 0.19 for white hens and between 0.0 and 0.10 for brown hens. Thus, it should be possible to select for A. galli resistance in chickens, which will be of importance for birds kept in alternative and organic farming systems.     

Effect and repeatability of Ascaridia galli egg output in cockerels following a single low dose infection

Three groups of caged 20 Tetra-SL cockerels aged 1 day were orally infected with 30, 60 or 125 embryonated Ascaridia galli eggs. After 11, 12 and 13 weeks, faecal egg counts (FECs) were determined. All birds were slaughtered after the last sampling. A group of 25 control birds was sampled and slaughtered in parallel. The gastrointestinal tracts were examined for the presence of adult stages of A. galli. A random sample of 10% was also examined for the presence of immature stages of A. galli. The group with an infection dose of 125 eggs showed the highest average worm burden (p<0.05) and number of females (p<0.05), but the mean establishment rate was the lowest in this group. There was no significant difference in the mean logFEC between the groups. The logFEC per female worm was the highest in the low infection group (p<0.05). The average worm length and weight and the birds body weight were not significantly different among the groups. The estimated repeatabilities for mean logFEC of the different samples were reasonably high (0.55-0.87). This may open a way of genetic selection for A. galli resistance in chickens, which will be of importance for birds kept in alternative and organic farming systems.     

Anthelmintic activity of ivermectin against experimental Ascaridia galli infection in chickens

The efficacy of ivermectin against Ascaridia galli infection was evaluated in chickens under controlled laboratory conditions in two separate experiments. In each experiment 22 White Leghorn chicks were randomly assigned to three groups of 10 (infected-treated), 9 (infected-control) and 3 (uninfected-control) birds. Each bird in the former two groups was orally infected with 1,500 embryonated A. galli eggs. The chicks in the treated group were subcutaneously injected with ivermectin at a dose of 0.3 mg kg-1 body weight on Day 10 (Experiment 1) and Day 35 post-infection (Experiment 2) for immature and adult infections, respectively. The treated birds had 0.9% (Experiment 1) and 0.4% (Experiment 2) worm recovery compared with 8.7 and 8% in the infected-untreated controls of the respective experiments. The fall in post-treatment faecal egg counts was 81 and 92% in birds treated on Days 10 and 35, respectively. The drug was found to be 90 and 95% effective against immature and adult worms, respectively. The lower lesion score and post-treatment near-normal haematobiochemical picture in treated birds confirmed these observations. The treated birds also had a better growth rate than the untreated chickens. The mature worms in the intestinal lumen of the host were more sensitive to the treatment than the immature stages of the parasite in the tissue phase.     

Establishment of Ascaridia galli in betamethasone-treated chickens

Twenty-five day-old White Leghorn chickens were each infected orally with 500 (Group I), 1000 (Group II) and 2000 (Group III) infective eggs of Ascaridia galli and were killed 30 days after the infection. A high percentage of the infecting dose (6.5%) established as adult worms in the intestine of chickens receiving the lowest level of primary infection, but as the amount of primary infection given to birds increased, there was a significant fall in the percentage establishment of adult worms in the intestine. A similar pattern of worm establishment was shown by chickens of the same age and receiving similar levels of primary infections, but which were treated with betamethasone at a dose of 2 mg/kg body weight commencing 5 days before and continuing up to 15 days after the infection. Betamethasone-treated birds, however, showed more establishment of worms in the intestine, but lower weight gains in comparison to the birds which were not treated. Different levels of primary infections given, with or without treatment with betamethasone, had no effect on the sex ratio of the resultant male/female worm populations, which became established in almost equal numbers in the intestine of chickens.     

The effect of Plasmodium gallinaceum on a challenge infection with Ascaridia galli in chickens

The effect of a primary infection with the haemoparasite Plasmodium gallinaceum on the establishment of a challenge infection with the nematode Ascaridia galli in chickens was studied. Four groups were infected as follows. Group 1: inoculated intravenously with 10(6) P. gallinaceum-infected erythrocytes on day 0; group 2: orally infected with 500 embryonated A. galli eggs on day 10; group 3: infected with P. gallinaceum on day 0 and A. galli on day 10; and group 3: non-infected control birds. The results of this investigation demonstrates that a primary infection with P. gallinaceum in chickens alters the course of a subsequent infection with A. galli. Thus, an antagonistic effect was seen in which the malaria infection caused a significant reduction on the establishment of the nematode in concurrently infected animals.     

Localization of Ascaridia galli larvae in the jejunum of chickens 3 days post infection

The normal habitat of the parasitic stages of Ascaridia galli is in the small intestine of poultry but the exact localization is poorly understood. Therefore, a histological study was conducted in order to localize the larvae during the early phase of infection. Six layer pullets seven-week old were infected orally with 20,000 embryonated A. galli eggs each, whereas four chickens were left as un-infected controls. At necropsy 3 days after infection the first half of jejunum/ileum was divided into two equally sized sections (J1 and J2). After taking samples for histology from the middle of J1 and J2 and the junction between these determined JX, the two sections were subjected to parasitological examination. A higher number of A. galli larvae were recovered from section J2 than J1 and the majority of larvae were recovered from the most profound layers. Based on histology 144 larvae were identified and their location was noted. The highest number of larvae was observed in the JX sample as compared to J1 and J2 (P<0.001). Most of them were located in the profound crypt zone of the mucosa (51%) as compared to the other zones (P<0.05). The number of larvae was higher in the lumen (63%) compared to the epithelium (32%) and lamina propria (5%) (P<0.001). A significantly higher number of eosinophils were found in lamina propria of the infected group compared to the control group (P<0.001). This experiment clearly showed that only few larvae had penetrated the epithelium and were positioned in the lamina propria at 3 days post infection. It was far more common that the larvae were localized within the epithelium or in the lumen of the crypts. It is therefore suggested that at least in this early phase "mucosal phase" is a more appropriate term to be used for the A. galli larval localization as compared to the term "histotrophic phase" currently used in many textbooks.     

Association study in naturally infected helminth layers shows evidence for influence of interferon-gamma gene variants on Ascaridia galli worm burden

ABSTRACT: Single nucleotide polymorphisms (SNPs) in the genes for interleukin-4, -13 and interferon-gamma, and 21 additional SNPs which previously had been significantly associated with immune traits in the chicken, were genotyped in white and brown layer hens and analyzed for their association with helminth burden following natural infections. A nucleotide substitution located upstream of the promoter of the interferon-gamma gene was significantly associated with the log transformed number of Ascaridia galli in the brown layer line (genotype CC: 6.4 ± 1.0 worms; genotype CT: 11.7 ± 2.2 worms). Therefore, IFNG seems to be a promising candidate gene for further studies on helminth resistance in the chicken.     

Effect of irradiated Ascaridia galli eggs on growth and cell-mediated immune responses in chickens

Growth and cell-mediated immune (CMI) responses were studied in 7-day-old chicks given orally 1000 irradiated (12.5 kR) or normal infective eggs of Ascaridia galli. Chicks immunised with irradiated eggs showed normal weight gains. CMI responses, as assessed by dinitrofluorobenzene (DNFB)-induced contact and delayed hypersensitivity reactions, were enhanced in the immunised group as compared with healthy controls, suggesting stimulation of CMI responses due to irradiation of A. galli eggs. CMI as well as growth responses were, however, found to be depressed in the birds administered normal infective eggs of A. galli. The present study highlights the role of the CMI response in protection against A. galli infection.     

Enrofloxacin assay validation and pharmacokinetics following a single oral dose in chickens

The pharmacokinetics of enrofloxacin (ENRO), a fluoroquinolone antimicrobial agent, was studied in male broiler chickens (Cobb) after single oral administration of 10 mg of ENRO/kg b.w. A high-performance liquid chromatography-photodiode array detector (DAD) (HPLC-DAD) method was developed and validated and used for quantitation of ENRO and its major metabolite ciprofloxacin in plasma. The HPLC analyses were carried out using a cationic-octadecyl mixed column and 0.05 mol/L phosphate buffer (pH 2.5)/acetonitrile as mobile phase. The sample preparation of plasma consisted of the precipitation of proteins followed by solid phase extraction on cationic-octadecyl mixed cartridges. The method was validated considering linear range, linearity, selectivity, sensitivity, limit of detection (LOD), limit of quantitation (LOQ), intra- and inter-day precisions and accuracy. The LOD and LOQ for both fluoroquinolones were 60 and 200 ng/mL for plasma. The plasma concentration vs. time graph was characteristic of a two-compartment open model. The maximal plasma concentration of 1.5 +/- 0.2 mg/mL was achieved at 9 +/- 2 h. The elimination half-life and the mean residence time of ENRO were 1.5 +/- 0.2 and 15.64 h, respectively. The area under the concentration-time curve was calculated as 35 +/- 4 mgxh/mL.     

The effect of concurrent infections with Pasteurella multocida and Ascaridia galli on free range chickens

Pasteurella multocida and Ascaridia galli are observed with high prevalences in free range chickens in Denmark, but the impact is unknown. A study was carried out to examine the interaction between A. galli and P. multocida in chickens and the impact on production.Five groups, each with 20 18-week-old Lohmann Brown chickens were infected. Group 1 was orally infected with 1000+/-50 embryonated A. galli eggs. Group 2 received 10(4) cfu P. multocida intratracheally. Group 3 was infected with A. galli and subsequently with P. multocida. Group 4 was infected with P. multocida followed by A. galli. Group 5 was the control. The study ran for 11 weeks where clinical manifestations, weight gain and egg production were recorded. Excretion of P. multocida was determined on individual basis and blood smears were made for differential counts. At the end of the study pathological lesions and the number of adult worms, larvae and eggs in the faeces were recorded.The birds were more severely affected when infected with both pathogens compared to single infections with A. galli or P. multocida, respectively. A lower weight gain and egg production was observed with dual infections. A. galli infection followed by a secondary P. multocida infection resulted in more birds with pathological lesions and continued P. multocida excretion.In conclusion a negative interaction between A. galli and P. multocida was observed and it is postulated that free range chickens are at higher risk of being subjected to outbreaks of fowl cholera when they are infected with A. galli.     

Comparison of parasite-specific immunoglobulin levels in two chicken lines during sustained infection with Ascaridia galli

Increasingly large numbers of poultry are held in production systems with access to outdoor areas. In these systems intestinal helminths are found with flock prevalences of up to 100%. Helminth infections influence chicken health negatively, which is why the following investigation has been performed.In the present experiment, 20 chickens of two inbred chicken lines containing the major histocompatibility complex (MHC) haplotypes, B14 and R5, were inoculated with 500 embryonated Ascaridia galli eggs. The A. galli-specific IgG titres of serum samples and the excretion of A. galli eggs in chicken faeces were measured for a period of 81 weeks.The level of excreted A. galli eggs measured as eggs per gram chicken faeces (EPG) varied greatly between chickens in each line. Significant differences were found between the two lines and with the R5 chickens reaching the highest levels. Likewise, the A. galli-specific IgG titres in serum differed significantly between the two lines, and an inverse relationship between infection level (EPG) and antibody titres was found. Although this inverse relationship suggests that humoral immunity may be involved in protection against A. galli infection, the high antibody titres did not prevent continued infection.     

Energy and nitrogen metabolism of diseased chickens: interaction of Ascaridia galli infestation and vitamin A status.

1. Effects of Ascaridia galli infection on the energy and nitrogen (N) metabolism were studied on groups of 5 cross-bred cockerels aged about 5 weeks and given a diet deficient or adequate in vitamin A at two levels of feeding in respiration chambers. 2. Metabolisability of dietary energy was 67% and N retention 33% in infected chickens compared with 71 and 41% respectively, in uninfected chickens. 3. Maintenance energy requirement of vitamin A-deficient birds was 882 kJ/kgW d compared with 998 kJ/kgW d for normal birds. N balance of the deficient chickens was also less when compared at the same energy balance. Infection did not affect maintenance energy requirement nor N balance. 4. Starvation heat production of infected chickens (619 kJ/kgW d) was higher than that of uninfected controls (586 kJ/kgW d). When infection treatments were combined, vitamin A-adequate chickens had a higher heat production (615 kJ/kg d) than the vitamin A-deficient (580 kJ/kgW d). Endogenous N excretion (mg/gW) was less in vitamin A-deficient than in adequate, starved birds. 5. Deficient chickens had undetectable liver reserves of vitamin A and only very low plasma concentrations. There was a difference in the length of larvae (17 d after infection) associated with vitamin A status, and with level of feeding.