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.     

Host age only partially affects resistance to primary and secondary infections with Ascaridia galli (Schrank, 1788) in chickens

Two experiments were conducted to compare the effect of chickens' age on resistance to primary and secondary infections with Ascaridia galli. In Experiment I, three groups, each of 80 female Lohman Brown chickens, aged one day, one month, or four months were compared. Within each group, 54 chickens were infected orally with 500 embryonated eggs and 26 were kept as non-infected controls. Weights were recorded weekly and five chickens in each group were slaughtered every 2 weeks for worm counts. At week 10 post-infection, 17 of the infected chickens and 18 of the controls were challenged with 500 eggs. In a replicate experiment (Experiment II), 35 one-day-old and 53 one-month-old female Lohman Brown chickens were infected orally with 500 A. galli eggs. Weights and fecal egg counts were recorded every week and infected chickens were necropsied every two weeks for determination of the worm burden. Chickens infected at one month of age excreted significantly fewer A. galli eggs when measured at 14 weeks of inoculation. The worms recovered from the one-month-old age group were significantly shorter than those from the chickens infected at one day of age in the first experiment. Worm burden and female fecundity values, however, were not significantly different between age groups in both Experiments I and II. Weight gains of infected chickens were not significantly different from the controls' and only a few chickens exhibited occasional slight diarrhea in both experiments. The results from these experiments demonstrate that the chickens' age only partially influences resistance to A. galli infection.     

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.     

Comparative genetic resistance to Ascaridia galli infections of 4 different commercial layer-lines

1. The objective of the study was to compare the establishment and effect of Asaridia galli infections in 4 different layer-lines. 2. A total of 160 birds comprising 4 different commercial layer-lines, ISA Brown, New Hampshire, Skalborg and a cross of New Hampshire(NH) and Skalborg (Sk), were infected with A. galli eggs. The birds were examined for the presence of parasite eggs and parasites at weeks 3, 6 and 9 post infection (pi). 3. At week 6 pi the chickens of the NH line harboured more larvae compared with the three otherlines. The Sk line chickens excreted more A. galli eggs throughout the study compared with the other lines. Female worms in the Sk line were more fecund than the worms in the other lines. Male and female worms recovered from the Sk line at week 9 pi were longer. Male worms recovered from the NH line 6 weeks pi were shorter than male worms from the other lines. Female worms recovered from the NH line were shorter than the female worms from the ISA line and the Sk line. No differences were seen in weight gain among the 4 lines. 4. The results suggest that genetic factors are involved in the establishment and survival of A. galli in the intestine of layers. Further studies are needed to elucidate the genetic mechanisms behind the observed parasitological findings.     

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.     

Ascaridia galli infections in free-range layers fed on diets with different protein contents

1. One hundred and twenty 17-week-old Lohman Brown hens were divided into 4 groups. Groups 1 and 3 were given a diet with 180 g protein/kg and groups 2 and 4 were given a diet with 140 g protein/kg. Groups 1 and 2 were orally infected with 500 (50) embryonated Ascaridia galli eggs. 2. Marked differences in mean weekly weight gain for the 4 groups were observed. 3. Hens given 140 g protein/kg had a significant lower mean worm burden of adult A. galli worms and a significant lower weight gain compared to the group given 18 g protein/kg. 4. There was no significant difference in faecal egg counts between the 2 parasitised groups. 5. The egg production did not differ significantly between any of the groups. 6. The results of this study indicate that the amount of dietary protein in the diet has an effect on the establishment of A. galli infections in the gut of layers kept under free range conditions.     

Flubendazole: safety regarding egg production and reproductive performance of breeder chickens

In a comparative field trial, 5,950 meat breeder chickens were medicated via feed for 7 consecutive days with 60 ppm of the anthelmintic flubendazole. This treatment cured the birds of an infection with Ascaridia galli and Capillaria obsignata. A slight but transitory diarrhea was observed in treated chickens. The applied medication did not unfavorably affect egg production, fertility, or hatchability.     

Genetic resistance to Ascaridia galli infections in chickens.

Ascaridiosis is still a cause of economic losses in modern poultry production systems. Improving genetic resistance to this disease provides an attractive alternative for free-range organic poultry producers and will eventually reduce the extensive use of anthelmintic drugs. An experiment was conducted to compare resistance to Ascaridia galli infections in Lohman Brown (LB) and Danish Landrace (DL) chickens. A self-cure mechanism to A. galli infections was observed in both breeds. However, significantly higher worm burdens and egg excretion were seen in the DL compared to LB chickens during primary infection. This indicates that breeding for resistance to A. galli is possible in chickens.     

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.     

Age-related differences of Ascaridia galli egg output and worm burden in chickens following a single dose infection

Ninety white chickens (Lohmann LSL) were reared under helminth-free conditions and divided into five groups. Four groups were artificially infected with 250 embryonated Ascaridia galli eggs at the age of 6, 12, 18 or 24 weeks. Ten birds were kept as uninfected controls. Six and 10 weeks after infection (p.i.), individual faecal egg counts (FEC) were performed. The birds were slaughtered after the second sampling and their gastrointestinal tracts were examined for the presence of adult A. galli. The FEC increased from the first to the second sampling significantly in all the infected groups. The highest increase was shown in the group infected at 12 weeks of age, whereas the increase in the other groups was relatively moderate. However, the total worm burden and mean FEC at the second sampling were highest (p<0.01) in those birds infected at an age of 12 or 18 weeks. The serum protein and triiodothyronine (T3) levels did not differ significantly (p>0.05) between any of the groups. Thyroxine (T4) was significantly different between the groups infected at 6 and 18 weeks of age (p<0.05), and those at 6 and 24 weeks of age (p<0.01). The thyroid hormone levels correlated significantly with the FEC. Age does not seem to play a major role in resistance to A. galli infections in layers, whereas a bird's hormonal and immune status, related to laying activity, seems to have a significant negative impact on resistance.     

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.     

Anthelmintic effects of citrus peels ethanolic extracts against Ascaridia galli

The use of phytogenic bioactive compounds to control poultry helminthes is increasing in different production systems. In vitro and in vivo anthelmintic activity of citrus peels against Ascaridia galli was investigated. Ethanolic extracts of three citrus peels species were suspended in 0.5% dimethyl sulfoxide (DMSO) to form an experimental composition (EC). EC was mainly composed of Limonene (96%), followed by β-Pinene (1.5%), α-Pinene (0.5%), and Sabinene (0.3%). For in vitro investigation, adult A. galli worms (n=225) were collected from naturally infected chickens and distributed to 3 equal groups. Groups 1, 2, and 3 were exposed to Fenbendazole (0.5mg/ml), EC (50mg/ml), and 0.5% DMSO, respectively. For in vivo investigation, 200 Lohmann Selected Leghorns chicks were infected at 1-day old with 250 embryonated A. galli eggs. At 6 weeks of age, 150 A. galli infected birds were randomly allocated into 5 equal groups. Groups 1, 2, and 3 were treated with 300, 600, and 1200 mg EC kg(-1) body weight, respectively. Group 4 was treated with Fenbendazole (50 mg kg(-1)). Group 5 was left as control. Birds were euthanized 2-weeks post-treatment, and all worms were collected from their intestines. EC possessed significant (P<0.001) in vitro anthelmintic properties on live worms. No significant (P>0.05) difference was quantified between number of motile worms exposed either to EC or Fenbendazole 7h post-exposure. A significant (P<0.0001) reduction in fecal egg count was observed 14 days post-treatment with 1200 mg kg(-1) EC. No significant differences were observed in worm burden of the 300 mg EC-treated group compared to the controls. In contrast, the 600 and 1200 mg EC-treated groups showed significant (P<0.0001) reduction in worm burden. Fenbendazole was the most effective in reducing A. galli burden (Efficacy=97%) followed by 1200 mg EC kg(-1) (68%), 600 mg EC kg(-1) (66%), and 300 mg EC kg(-1) (5%). It is concluded that citrus peels extracts have potential anthelmintic properties against A. galli.     

Response of two breeds of chickens to Ascaridia galli infections from two geographic sources

Comparative resistance to different isolates of Ascaridia galli was investigated in a local chicken breed from Jordan (LC) and in the Lohmann LSL white chicken (LW) strain. In two trials, birds of LC and LW were inoculated orally at 1-day old with 250 embryonated A. galli eggs. In the first trial a German source of A. galli was used, whereas in the second trial, a Jordan source of A. galli was used. At week 7 of infection, infected LC birds harbored significantly (P<0.05) fewer worms and excreted less A. galli eggs than infected LW birds. A. galli isolated from Jordan were less infectious than A. galli from Germany. Results suggest that the variation in genetic background between LC and LW is involved in the resistance to A. galli infection. A. galli isolates from different geographic areas differ in their ability to infect different chicken genotypes.     

Pasteurella multocida infection in heterophil-depleted chickens

The present study was aimed at elucidating the role of heterophil granulocytes during the initial infection with Pasteurella multocida subsp. multocida in chickens. Chickens (17 and 19 wk old) were depleted of their heterophil granulocytes by 5-fluorouracil treatment. When the heterophil blood counts were significantly reduced, the birds were inoculated intratracheally with 1.8-4.3 x 10(4) colony-forming units of P. multocida. Twelve, 24, or 48 hr postinoculation, the birds were euthanatized and examined for macroscopic and histologic lesions in the lungs. Bacterial invasion was determined by culture of P. multocida from the spleen. Recruitment of heterophils into the respiratory tract during infection was found to contribute considerably to the lung lesions in chickens and was found to mediate tissue damage, possibly allowing a more rapid systemic spread of P. multocida. However, during progression of the infection, the heterophil-mediated necrosis in chickens seemed to stimulate giant cell demarcation of infected lung tissue, which coincided with the clearance of P. multocida from the spleen, thus hampering further invasion. Consequently, heterophil activation plays a dual role for the outcome of a P. multocida infection in chickens, where it initially seems to promote invasion and systemic spread but subsequently helps limit the infection by giant cell formation and bacterial clearance.     

Influence of Ascaridia galli infections and anthelmintic treatments on the behaviour and social ranks of laying hens (Gallus gallus domesticus)

In the present study, the effects of an experimental Ascaridia galli infection and anthelmintic treatment on the behaviour and social status of laying hens of two different lines were studied. Sixty white (Lohmann LSL; LSL) and 60 brown (Lohmann Brown; LB) hens were reared under helminth-free conditions. The hens of each line were divided into four groups. The birds in two of the groups were artificially infected with 250 embryonated A. galli eggs at an age of 27 weeks. The other two groups were kept as uninfected controls. One infection and control group was dewormed at 38 weeks of age and slaughtered 4 weeks later, contemporary with the other animals. Individual faecal Ascaridia egg counts (FEC) were performed 11 weeks post-infection (p.i.). Body weights, laying performance and egg weights were recorded regularly. Blood was taken to measure testosterone levels. The worm burdens established in the intestines were counted in the infected not treated group after slaughtering. In addition, 15 behavioural parameters were recorded by focal animal observation (n=10 per group) of one infection (plus anthelmintic treatment) and one control group, according to the time-sampling method throughout the experiment. All agonistic interactions within the groups were recorded simultaneously on an ongoing basis, thereby allowing the calculation of an individual social rank index. The following results were obtained: Mean FEC and worm burden were higher (p < 0.01) in the LSL hens than in the LB hens, but their performances were not different (p > 0.05) from the controls. Infections with A. galli resulted in significant behavioural changes in both lines as the infected birds showed a higher food intake and lower locomotion activity during the prepatent and patent periods. After anthelmintic treatment, food intake decreased and locomotion increased. Behavioural changes were more pervasive in the infected LSL hens, as these hens also showed changes in ground pecking and nesting activity not only during the prepatent and patent periods, but also after anthelmintic treatment. Social rank did not significantly change as a consequence of A. galli infection, but the infection groups of both lines had a tendency to display more agonistic activity than the non-infected controls. Serum testosterone values only increased significantly in the LSL hens during the patent period compared to the controls. No significant correlations were apparent between serum testosterone level, parasitic parameters, social status and/or aggressiveness in any of the groups. This study showed that even sub-clinical A. galli infections and their anthelmintic treatment can have an impact on animal behaviour.     

Fowl adenovirus (FAdV) serotype 4 causes depletion of B and T cells in lymphoid organs in specific pathogen-free chickens following experimental infection

In the present investigation flow cytometric analysis and immunohistochemistry were applied together for the first time to gain new insights into the interaction between virulent fowl adenoviruses (FAdV) and the immune system of chickens. As a model for virulent FAdV infections a FAdV-4 strain was used, known as the aetiological agent of Hepatitis-Hydropericardium syndrome (HHS) in broilers sometimes also named Angara Disease. Specified pathogen-free chickens (SPF) were divided into three different groups. Group I was infected at first day of life with an attenuated form of the virus obtained through continuous cell culture passage with the virulent virus and then re-infected 3 weeks later with the virulent progenitor virus. Group II was solely infected with the virulent virus at 3 weeks and group III served as a negative control. Following infection with the virulent virus a decrease of CD3+, CD4+ and CD8+ cells was noticed in the spleen. This was accompanied by a decrease of CD4+ and CD8+ T-lymphocytes in the thymus. Those birds infected with the attenuated virus in first instance and challenged with the virulent virus did not show these pathological effects in the thymus. In the bursa of Fabricius a severe depletion of lymphocytes was observed by immunohistochemistry in birds, infected with the virulent virus. Taken together it can be concluded that an infection with FAdV-4 has profound effects on cells, of the humoral and cell-mediated immune responses. The effects are much more severe in the birds infected with the virulent virus only indicating that the preceding infection with the attenuated virus reduces significantly the adverse effects induced by the virulent virus.     

The response of broiler breeder chickens to parenteral administration of avirulent Pasteurella multocida

Broiler breeder chickens were exposed to avirulent Pasteurella multocida at 14, 22, and 34 weeks of age either by stick wing 1 to 3 times or subcutaneously 3 times. Fowl pox vaccine was mixed with the first P. multocida exposure in some groups. Exposure did not impair egg production or hatch of fertile eggs. Challenge with pathogenic P. multocida serotype 1 at 68 weeks indicated that exposure to avirulent P. multocida 2 or 3 times provided better protection than 1 exposure. Mixing fowl pox vaccine with the avirulent P. multocida did not reduce immunity to fowl cholera or fowl pox.     

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.     

Treatment of experimentally induced Pasteurella multocida infections in broilers and turkeys: comparative studies of different oral treatment regimens

Experimental fowl cholera was induced in 60 healthy 10-week-old broiler chickens and 8-week-old turkeys by intramuscular inoculation with approximately 80 colony-forming units (cfu) of Pasteurella multocida X-73 strain and with approximately 70 cfu of P. multocida P-1059 strain, respectively. This method of infection proved to be useful for evaluating the efficacy of anti-microbial medication, by measuring mortality, weight gain, pathological responses and frequency of re-isolation of P. multocida. The efficacies of two different dosing methods, continuous and pulse dosing, were compared. Using the continuous-dosing method, norfloxacin was administered to drinking water at 100 mg/l for 5 days in chickens. Efficacies were slightly improved compared with pulse dosing at 15 mg/kg bodyweight for the same length of time. The opposite was observed in turkeys, to the degree of control of mortality and maintenance of weight gain.     

Detection of antibodies in serum and egg yolk following infection of chickens with an H6N2 avian influenza virus.

Active serologic surveillance programs to detect avian influenza viruses (AIVs) in table egg-laying chickens have been initiated by several states as a response to the economic threat posed by these viruses. Most outbreaks of avian influenza in domestic poultry are caused by mildly pathogenic AIVs. In the study reported here, infection by an H6N2 AIV was used as a model of mildly pathogenic AIV infections in egg-type chickens. The total number of eggs laid by 5 control hens was 619 or 0.904 eggs/day/hen, whereas the total number laid by 10 infected hens was 1,018 or 0.743 eggs/day/hen. The difference in egg production between the 2 groups was not statistically significant (P = 0.38). Anti-influenza antibodies were monitored by use of an agar gel immunodiffusion test and an ELISA for a period of 20 weeks after inoculation. Antibodies in serum developed sooner, peaked at higher levels, and remained at higher levels than did antibodies found in egg yolk, as indicated by ELISA results. For infected chickens, the correlation between serum and egg yolk ratios was 0.66. Serum samples would appear to be preferable to egg yolk samples for surveillance programs intended to identify chicken flocks that may have been infected by an AIV weeks or months before samples are collected.