Infection with Ascaridia galli Does Not Significantly Alter Intestinal Microbiota and Is Cleared After Changes in the Expression of Cytokines.

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-ß, 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.

La infección por Ascaridia galli no altera significativamente la microbiota intestinal y se elimina tras cambios en la expresión de citocinas. Debido a la tendencia de la producción de huevos libres de jaulas, las infecciones con el nematodo Ascaridia galli están recibiendo una mayor atención. El objetivo de este estudio fue establecer una línea de tiempo para la influencia de A. galli en la expresión de citoquinas clave relacionadas con una respuesta inmune parasitaria y en la composición de la microbiota yeyunal. Veintiocho aves macho de tipo postura fueron desafiadas a los 24, 25 y 26 días de edad. Se mantuvieron 28 aves adicionales como controles no infectados. A partir del día 31, se practicó la eutanasia a tres aves de cada grupo cada semana hasta las 8 semanas posteriores a la infección (PI). El número de larvas aisladas de la pared intestinal disminuyó con el tiempo, hasta que no se observaron larvas a las 7 y 8 semanas después de la infección. A las cinco semanas post-infección, hubo una regulación ascendente numérica de todas las citoquinas (TGF-ß, IFN-γ, IL-4, IL-8, IL-10, IL-13) en el grupo infectado, pero este cambio solo fue estadísticamente significativo para IL-13. En ese momento, se esperaba que las larvas se hubieran convertido en adultos que eliminarían huevos en las heces. Sin embargo, no se observaron nemátodos adultos y no hubo eliminación de huevos. Para el análisis de microbiota, hubo diferencias significativas en la diversidad alfa (diversidad filogenética de Faith) entre los grupos de desafío y control y el análisis de diversidad beta mostró ligeras diferencias entre las muestras, lo que sugiere que la edad de las aves fue la razón principal de la separación de los grupos. Estos hallazgos sugieren que la regulación al alza de todas las citocinas evaluadas en la semana 5 podría ser el motivo de la resolución de la infección. Las posibles explicaciones son que una dosis alta de infección y el hecho de que las aves fueran alimentadas con un alimento más denso desde el punto de vista nutricional podrían haber contribuido a que el sistema inmunitario de las aves eliminara la infección antes de que los nemátodos pudieran alcanzar la madurez.

The impacts of Ascaridia galli on performance, health, and immune responses of laying hens: new insights into an old problem.

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.

Analysis of antibody levels in egg yolk for detection of exposure to Ascaridia galli parasites in commercial laying hens.

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.

Co-expulsion of Ascaridia galli and Heterakis gallinarum by chickens.

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.

Time- and dose-dependent development of humoral immune responses to Ascaridia galli in experimentally and naturally infected chickens.

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.

Effect of an artificial Ascaridia galli infection on egg production, immune response, and liver lipid reserve of free-range laying hens.

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.

Immune responses following experimental infection with Ascaridia galli and necrotic enteritis in broiler chickens.

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.

Maternal protection against Ascaridia galli?

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.

Immune gene expression in the spleen of chickens experimentally infected with Ascaridia galli.

Ascaridia galli is a gastrointestinal nematode infecting chickens. Chickens kept in alternative rearing systems or at free-range experience increased risk for infection with resulting high prevalences. A. galli infection causes reduced weight gain, decreased egg production and in severe cases increased mortality. More importantly, the parasitised chickens are more susceptible to secondary infections and their ability to develop vaccine-induced protective immunity against other diseases may be compromised. Detailed information about the immune response to the natural infection may be exploited to enable future vaccine development. In the present study, expression of immune genes in the chicken spleen during an experimental infection with A. galli was investigated using the Fluidigm(®) BioMark™ microfluidic qPCR platform which combines automatic high-throughput with attractive low sample and reagent consumption. Spleenic transcription of immunological genes was compared between infected chickens and non-infected controls at week 2, 6, and 9 p.i. corresponding to different stages of parasite development/maturation. At week 2 p.i. increased expression of IL-13 was observed in infected chickens. Increased expression of MBL, CRP, IFN-α, IL-1ß, IL-8, IL-12ß and IL-18 followed at week 6 p.i. and at both week 6 and 9 p.i. expression of DEFß1 was highly increased in infected chickens. In summary, apart from also earlier reported increased expression of the Th2 signature cytokine IL-13 we observed only few differentially expressed genes at week 2 p.i. which corresponds to the larvae histotrophic phase. In contrast, we observed increased expression of pro-inflammatory cytokines and acute phase proteins in infected chickens, by week 6 p.i. where the larvae re-enter the intestinal lumen. Increased expression of DEFß1 was observed in infected chickens at week 6 p.i. but also at week 9 p.i. which corresponds to a matured stage where adult worms are present in the intestinal lumen.

Ascaridia galli infection influences the development of both humoral and cell-mediated immunity after Newcastle Disease vaccination in chickens.

Potent vaccine efficiency is crucial for disease control in both human and livestock vaccination programmes. Free range chickens and chickens with access to outdoor areas have a high risk of infection with parasites including Ascaridia galli, a gastrointestinal nematode with a potential influence on the immunological response to vaccination against other infectious diseases. The purpose of this study was to investigate whether A. galli infection influences vaccine-induced immunity to Newcastle Disease (ND) in chickens from an MHC-characterized inbred line. Chickens were experimentally infected with A. galli at 4 weeks of age or left as non-parasitized controls. At 10 and 13 weeks of age half of the chickens were ND-vaccinated and at 16 weeks of age, all chickens were challenged with a lentogenic strain of Newcastle disease virus (NDV). A. galli infection influenced both humoral and cell-mediated immune responses after ND vaccination. Thus, significantly lower NDV serum titres were found in the A. galli-infected group as compared to the non-parasitized group early after vaccination. In addition, the A. galli-infected chickens showed significantly lower frequencies of NDV-specific T cells in peripheral blood three weeks after the first ND vaccination as compared to non-parasitized chickens. Finally, A. galli significantly increased local mRNA expression of IL-4 and IL-13 and significantly decreased TGF-ß4 expression in the jejunum two weeks after infection with A. galli. At the time of vaccination (six and nine weeks after A. galli infection) the local expression in the jejunum of both IFN-? and IL-10 was significantly decreased in A. galli-infected chickens. Upon challenge with the NDV LaSota strain, viral genomes persisted in the oral cavity for a slightly longer period of time in A. galli-infected vaccinees as compared to non-parasitized vaccinees. However, more work is needed in order to determine if vaccine-induced protective immunity is impaired in A. galli-infected chickens.

Ascaridia galli infection affects pullets differently when feed is contaminated with the Fusarium toxin deoxynivalenol (DON).

The Fusarium mycotoxin deoxynivalenol (DON) is a common contaminant of cereal grains used as animal feed. DON is known for its cytotoxic and anti-proliferative properties and might adversely affect the health of poultry. The prevalence of the intestinal parasitizing roundworm Ascaridia galli is higher in outdoor housing systems and has been associated with maldigestion and malabsorption. It was hypothesized that ingested DON might not only affect the pullet itself but could also act on the nematode parasitizing in the ingesta. To examine these interactions between A. galli infection and DON contamination of feed 4 groups of 9 pullets in each were tested; non-infected groups were fed either an uncontaminated control (CON-) or a Fusarium toxin contaminated and mainly DON-containing diet (FUS-), and the corresponding A. galli inoculated groups were fed accordingly (CON+, FUS+). A. galli infection significantly reduced the jejunal villi height and increased the thickness of the tunica muscularis with the effect being more pronounced when the DON-containing diet was fed (Group FUS+). Only in this group significantly increased weights of jejunal and ileal tissues and of livers were noticed. Moreover, DON was detected in plasma of the pullets at higher frequencies when they were infected suggesting a facilitated absorption of DON. Group FUS+ was characterized by a significantly higher excretion of A. galli eggs and a concomitant lower proportion of pullets with detectable antibodies against a somatic antigen of A. galli while worm burden and worm characteristics were not affected by diet. Other effects of feeding the FUS diet to the infected pullets included an increased mass per length of male worms. In conclusion, infection of pullets with A. galli might increase the susceptibility towards DON as indicated by an increased DON absorption rate and a compromised antibody formation. The effects of DON on fecundity and worm morphology require further examination.

Proteomic analysis of Ascaridia galli. Identification of immunoreactive proteins in naturally and experimentally infected hens.

Ascaridia galli, intestinal parasite of domestic fowl, is responsible of economic losses in avian exploitations. However, molecular mechanisms that govern avian ascaridiasis remain largely unknown. The aim of the present work was to identify proteins of A. galli recognized by the immune system of naturally and experimentally infected hens, using two-dimensional electrophoresis (2-DE) and mass spectrometry (MS). Sixteen immunoreactive proteins of A. galli were identified. These proteins are mainly related to different metabolic processes, cell motility and binding activities. The timing evolution of this recognition pattern was studied using serum samples from experimentally infected hens, allowing us to observe an early recognition of many of these antigens. Many of them were isoforms from lipid and plasminogen-binding proteins. Moreover, plasminogen-binding activity has been related in other parasites with the facilitation of intra-organic migration, which represents an important fact in avian ascaridiasis. This work represents the first proteomic study of A. galli and could contribute to explain some aspects of parasite/host relationships of avian ascaridiasis.

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.

Immunopathogenesis of Ascaridia galli infection in layer chicken.

Gastro-intestinal nematode infections in mammals are associated with local T lymphocyte infiltrations, Th2 cytokine induction, and alterations in epithelial cell secretion and absorption. This study demonstrates that Ascaridia (A.) galli infection in chicken also elicits local gut-associated immune reactions and changes in the intestinal electrogenic nutrient transport. In A. galli-infected birds we observed infiltrations of different T cell populations in the intestinal lamina propria and accumulation of CD4+ lymphocytes in the epithelium. The Th2 cytokines IL-4 and IL-13 dominated the intestinal immune reactions following A. galli infection. A. galli-specific systemic IgY antibodies were detected after two weeks post infection, and did only poorly correlate with detected worm numbers. Electrogenic transport of alanin and glucose was impaired in A. galli-infected chicken. Our data provide circumstantial evidence that local immune responses and electro-physiological intestinal functions may be connected and contribute to the elimination of worm infection.

MHC haplotype and susceptibility to experimental infections (Salmonella Enteritidis, Pasteurella multocida or Ascaridia galli) in a commercial and an indigenous chicken breed.

In three independent experimental infection studies, the susceptibility and course of infection of three pathogens considered of importance in most poultry production systems, Ascaridia galli, Salmonella Enteritidis and Pasteurella multocida were compared in two chicken breeds, the indigenous Vietnamese Ri and the commercial Luong Phuong. Furthermore, the association of the Major Histocompatibility Complex (MHC) with disease-related parameters was evaluated, using alleles of the LEI0258 microsatellite as markers for MHC haplotypes. The Ri chickens were found to be more resistant to A. galli and S. Enteritidis than commercial Luong Phuong chickens. In contrast, the Ri chickens were more susceptible to P. multocida, although production parameters were more affected in the Luong Phuong chickens. Furthermore, it was shown that the individual variations observed in response to the infections were influenced by the MHC. Using marker alleles of the microsatellite LEI0258, which is located within the MHC region, several MHC haplotypes were identified as being associated with infection intensity of A. galli. An association of the MHC with the specific antibody response to S. Enteritidis was also found where four MHC haplotypes were shown to be associated with high specific antibody response. Finally, one MHC haplotype was identified as being associated with pathological lesions and mortality in the P. multocida experiment. Although not statistically significant, our analysis suggested that this haplotype might be associated with resistance. These results demonstrate the presence of local genetic resources in Vietnamese chickens, which could be utilized in breeding programmes aiming at improving disease resistance.

Antibody and inflammatory responses in laying hens with experimental primary infections of Ascaridia galli.

Ascaridia galli, an intestinal nematode that affects hens and other domestic and wild birds, causes economic losses in avian exploitations. The present work shows that A. galli stimulates a strong antibody response as well as an intense inflammatory reaction, in the intestinal mucous of experimentally infected Lohmann Brown laying hens. IgG antibodies against soluble extracts of A. galli embrionated eggs and adult worms, were detected in both blood and yolks eggs from infected hens during a period of 105 days after the infection. This indicates that hens transfer to their offspring a part of the IgG antibodies produced when they become infected. The antigens responsible for the stimulation of specific IgG were molecules of 30-34, 44-54 and 58-90 kDa, while in the yolk eggs of infected hens a reactivity directed against antigens of molecular weight (M(w)) lower than 50 kDa was detected. Histology revealed traumatic lesions with leukocyte infiltration, and inflammation of the intestinal wall of the infected hens after 105 days of initial infection. The possible influence of the immune and inflammatory response on the population dynamics of the parasite is discussed.

Case of retinochoroiditis associated with porcine ascarids diagnosed by antibodies in subretinal fluid.

PURPOSE: To determine the cause of retinochorioditis in a patient with a granulomatous retinal exudate and an exudative retinal detachment. CASE: A 45-year-old man presented at another hospital with increased visual disturbances of the left eye. He was diagnosed with uveitis, and treated with topical steroids for 1 month. However, the uveitis worsened, and he was referred to our hospital. Ophthalmoscopy showed a yellowish-white granulomatous exudate, and an exudative retinal detachment in the lower peripheral retina. The retinal detachment worsened and affected the macula. Pars plana vitrectomy was performed, and the retina was reattached. During the surgery, ocular samples were collected for further examinations. The titers of antibodies against 12 kinds of ascaridis were examined, and elevated titers of specific antibodies against porcine ascarids were detected in the subretinal fluid, but not in the aqueous humor, vitreous, or serum. CONCLUSIONS: Vitrectomy with the collection of ocular samples, especially subretinal fluid, was a key procedure in the diagnosis and treatment of retinochoroiditis associated with the porcine ascarids.

Th1/Th2 polarization by viral and helminth infection in birds.

Mammals developed an immune system able to functionally polarize into so-called type 1 or type 2 immune pathways, to resolve infections with intracellular and extracellular pathogens, respectively. In the well-studied avian immune system of the chicken, however, no evidence for polarized immunity could be found, as yet. To investigate whether these two major arms of mammalian immunity, regulated by a T helper (Th)1/Th2 cytokine balance, evolved similarly in birds, chickens were exposed to a prevalent intracellular (viral) or extracellular (helminth) infection. By using semi-quantitative RT-PCR analysis we provide evidence that polarization of Th1/Th2 type immunity extends beyond mammalian species, and, therefore, has been evolutionary conserved for more than 300 million years, when the lineages of mammalian and avian vertebrates are assumed to have segregated.

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.

Effects of glycine-metal compounds on Ascaridia galli-infected chickens expressed by a kinetic model.

The biogenic elements zinc, manganese and cobalt are essential for metabolic processes in animals. Compounds of nGly.Me2+A. mH2O (Me2+=Zn2+, Mn2+, Co2+; A=Cl(-), SO4(2-), n=1, 2; m=2, 5), as supplements in the diet, were used separately on different experimental groups of male Hisex chickens to correct the mineral deficiency caused by Ascaridia galli infections. An amelioration of body weight gain, reduction of mortality and restoration of trace element levels were estimated in infected chickens. A mathematical model has been proposed for A. galli population kinetics in chickens, taking into account the stimulating effect of these elements on the nematodes. The model parameters are considered as phenomenological constants of the host-parasite system. An agreement with experimental data is observed using, for the parameters psi, alpha, micro and micros, values equal to those calculated in previously investigated A. galli-chicken systems. For parameter nu (immunological constant) the same value was obtained as in a previous experiment with high infection. This model is likely to be suitable for a range of host-nematode systems, including varying degrees of infection and treatment with different trace elements.