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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.
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Anti-Helmínticos , Ascaridia , Ascaridíase , Galinhas , Juglans , Extratos Vegetais , Folhas de Planta , Doenças das Aves Domésticas , Animais , Ascaridia/efeitos dos fármacos , Extratos Vegetais/farmacologia , Folhas de Planta/química , Juglans/química , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/parasitologia , Anti-Helmínticos/farmacologia , Ascaridíase/veterinária , Ascaridíase/tratamento farmacológico , Ascaridíase/parasitologia , Etanol/química , Etanol/farmacologiaRESUMO
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 5â¯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.
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Ascaridia , Ascaridíase , Galinhas , Técnicas de Amplificação de Ácido Nucleico , Doenças das Aves Domésticas , Sensibilidade e Especificidade , Animais , Galinhas/parasitologia , Técnicas de Amplificação de Ácido Nucleico/veterinária , Técnicas de Amplificação de Ácido Nucleico/métodos , Doenças das Aves Domésticas/parasitologia , Doenças das Aves Domésticas/diagnóstico , Ascaridia/isolamento & purificação , Ascaridia/genética , Ascaridíase/veterinária , Ascaridíase/diagnóstico , Ascaridíase/parasitologia , Fezes/parasitologia , Técnicas de Diagnóstico Molecular/veterinária , Técnicas de Diagnóstico Molecular/métodosRESUMO
The periodicity of parasite egg excretion refers to variations in the number of eggs produced across time, with significant implications in optimizing diagnostic procedures and conducting the Fecal Egg Count Reduction Test (FECRT). Here, we explore whether Ascaridia galli egg excretion varies across time under Philippine conditions, thus informing the best time to collect fecal samples during flock health examination. A time-course analysis was performed in chickens (N = 12) experimentally infected with A. galli, isolated from a naturally infected Philippine native chicken. We examined the fecal egg per gram (EPG) count at 3-h intervals for 3 days, starting from 5:00-6:00 h AM to the following day at 1:00-2:00 h AM. Our results showed a consistent daily egg excretion pattern with a peak EPG count in the morning that abruptly declined in the afternoon and lowest in the evening. The EPG counts correlated with the amount of excreta produced, suggesting that A. galli fecundity corresponds to the timing of host defecation. Our results imply that the best time to collect fecal samples for A. galli diagnosis and FECRT in Philippine conditions should be from sunrise until late morning when parasite EPG count and host excreta production are at their highest.
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Colorimetric assays are some of the most convenient detection methods, creating discoloration in solutions that is visible to the naked eye. However, colorimetric reactions have some limitations regarding the variability in the color perception of individuals caused by factors such as color blindness, experience, and gender. Semi-quantitative chromatic analysis has been used as an alternative method to differentiate between two colors and accurately interpret the results from a numerical value, with high confidence. Therefore, we developed and determined the optimal model between Red-Green-Blue (RGB) and Commission Internationale de l'Eclairage (CIE) Lab color spaces to establish a semi-quantitative colorimetric assay via image analysis by the ImageJ program for loop-mediated isothermal amplification (LAMP), using the dyes malachite green and phenol red. The semi-quantitative colorimetric assays using the color distance values of the CIELab color space (ΔEab) were more suitable than those using the RGB color space (ΔERGB) for chromatic differentiation between positive and negative reactions in both indicator dyes, demonstrating the feasibility of this assay to be applied in the detection of a wide range of pathogens and infectious diseases.
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Colorimetria , Técnicas de Amplificação de Ácido Nucleico , Humanos , Colorimetria/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Corantes , Técnicas de Diagnóstico MolecularRESUMO
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.
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Ascaridia , Ascaridíase , Humanos , Animais , Feminino , Ascaridia/fisiologia , Ascaridíase/veterinária , Ascaridíase/parasitologia , Galinhas , Imunidade , FígadoRESUMO
Ascaridia galli (Nematoda: Ascaridiidae), infecting mainly the small intestine of chickens, is one of the most common nematodes in poultry worldwide. The complete mitochondrial genome sequence of A. galli was 13,981 bp in total length with 36 coding genes, namely, 12 protein-coding genes (PCGs), two ribosomal RNAs, and 22 transfer RNAs. All PCGs were transcribed in one direction. Phylogenetic analysis of the mitogenome of A. galli would further contribute to resolving its phylogenetic position and offer novel perspectives on phylogenetic studies of A. galli.
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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-ß4 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.
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Ascaridia , Ascaridíase , Animais , Feminino , Ascaridíase/veterinária , Ascaridíase/parasitologia , Galinhas/parasitologia , Citocinas , Mucosa Intestinal , AnticorposRESUMO
Ascaridia galli is an important nematode that causes ascaridiasis in free-range and indoor system chicken farms. Infection with A. galli may damage the intestinal mucosa and inhibit nutrient absorption, leading to a reduced growth rate, weight loss and a decreased egg production. Consequently, A. galli infection is a significant health problem in chickens. In this study, we developed a loop-mediated isothermal amplification coupled with a lateral flow dipstick (LAMP-LFD) assay for the visual detection of A. galli eggs in faecal samples. The LAMP-LFD assay consists of six primers and one DNA probe that recognize the internal transcribed spacer 2 (ITS2) region; it can be performed within 70â min and the results can be interpreted with the naked eye. Using the LAMP-LFD assay developed in this study, A. galli DNA was specifically amplified without any cross-reactions with other related parasites (Heterakis gallinarum, Raillietina echinobothrida, R. tetragona, R. cesticillus, Cotugnia sp., Echinostoma miyagawai) and definitive hosts (Gallus gallus domesticus, Anas platyrhynchos domesticus). The minimum detectable DNA concentration was 5 pg/µl, and the detectable egg count was 50 eggs per reaction. The assay can be performed in a water bath, without the need for post-mortem morphological investigations and laboratory instruments. It is therefore a viable alternative for the detection of A. galli in chicken faeces and can replace classical methods in field screening for epidemiological investigations, veterinary health and poultry farming management.RESEARCH HIGHLIGHTSThis is the first study using the LAMP-LFD assay for Ascaridia galli detection.The results can be observed by the naked eye.The developed assay can be used to detect Ascaridia galli eggs in faecal samples.
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Ascaridia , Galinhas , Animais , Estudos de Viabilidade , Galinhas/parasitologia , Óvulo , Fezes/parasitologia , DNARESUMO
The present study aims to find efficient alternatives to synthetic anthelmintics among ethno-veterinary herbs. Ascaridia galli eggs isolated from the worm uterus were exposed in vitro to methanolic extracts (ME) of nine plant species such as Achillea millefolium (AM), Artemisia absinthium (AA), Artemisia vulgaris (AV), Cicerbita alpina (CA), Cichorium intybus (CI), Inula helenium (IH), Origanum vulgare (OV), Tanacetum vulgare (TV), Tanacetum parthenium (TP). Flubendazole (FL), 0.5% formalin with dimethylsulfoxide and Petri dishes without the addition of reagents were used as positive, negative and untreated control respectively. The effects of the different ME at concentrations 0.500, 0.325, 0.200 mg/ml were assessed on the embryonic development (ED) of the eggs in duplicate. Logit analysis was used to calculate EC50 values. A generalized linear mixed model, having plant species and concentration as fixed effect and day as repeated measure, was used to determine differences in ED. Estimated EC50 was the lowest for FL at 0.11 mg/ml. CA and TV followed with 0.27 mg/ml and 0.32 mg/ml. ED for FL was significantly lower (25%) than that of CA (47%). The analysis showed 0.5 mg/ml of the ME of CA and TV significantly affected the ED at 35% and 42% inhibitions respectively. The ED for all ME showed similar pattern i.e., relatively higher efficacy in the first experimental week compared to the rest of the experimental period. The effect from all multicomponent extracts is time and dose dependent. The plants have promising results in inhibiting ED, contributing to the identification of alternative anthelmintic treatments.
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Ascaridia , Mebendazol , Animais , Feminino , Dimetil Sulfóxido , Formaldeído , MetanolRESUMO
Helminth parasite infections are widespread in smallholder farming systems affecting farmers and livestock animals. There are pathogenic parasites that populate the gut of their host and coexist closely with the gut microbiota. The physical and immunological environment of the gut can be modified by parasites and microbiota creating a wide range of interactions. These interactions modify the development of infection, affects overall host health, and can modify the way a host interacts with its bacterial microbiota. In addition, where there is a high worm burden parasites will affect the health of the host and intestinal tract colonization. This review highlights key studies on the interaction between helminth parasites and the intestinal microbiome to understand the relationship between parasitic worm infections and gut microbiome health in chickens. Finally, the review discusses modulations, molecular changes, and the importance of helminth-microbiome interactions for the host.
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Free-range chickens are an integral part of poultry production in many developing countries. In the Mountain Kingdom of Lesotho, the majority of the population own free-range chickens, which serve a variety of purposes including being a source of meat, eggs and use for cultural rituals amongst others. However, there is lack of scientific studies on occurrence of parasitic infections on free-range chickens in Lesotho. The aim of this study was to document common gastrointestinal parasites infecting free-range chickens in four villages of Mafeteng District in Lesotho. A total number of 462 pooled faecal samples were collected from various households in HaKubutu (n = 114), HaMatjeka (n = 120), HaMpalipali (n = 120) and Thabang Villages (n = 108) which were subjected to microscopic examination using McMaster technique. The prevalence of gastrointestinal parasite infection was as follows: Eimeria tenella (12.8%), Ascaridia galli (10.4%) and Heterakis gallinarum (5%). The prevalence for H. gallinarum and Ascaridia galli were comparatively higher during the hot-wet season (7.1% and 2.8% respectively) than the cold-dry season (3.2% and 1.9% respectively) and varied significantly (P < 0.05). For E. tenella, the oocysts per gram were slightly higher in the cold-dry season than the hot-wet season. Polymerase chain reaction only amplified DNA from six (29%) adult A. galli worms of which two amplicons were successfully sequenced. The obtained cytochrome C oxidase subunit 1 partial gene sequences displayed 98-100% identity with South African A. galli isolates. This is the first scientific study on prevalence and molecular characterization of nematodes and coccidia species infecting free-range village chickens in Lesotho. The findings can be used to review management of gastrointestinal nematodes and protozoal parasites of free-range chickens in Lesotho.
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Coccídios , Parasitos , Animais , Galinhas , Lesoto/epidemiologia , AscaridiaRESUMO
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.
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Ascaridíase , Doenças das Aves Domésticas , Animais , Coelhos , Galinhas , Ascaridíase/diagnóstico , Ascaridíase/veterinária , Doenças das Aves Domésticas/diagnóstico , Óvulo , Ascaridia , Ensaio de Imunoadsorção Enzimática/veterináriaRESUMO
Background and Aim: Roundworms cause infections in the avian population that lead to illness and poor production. The singing zebra dove is an economically important animal in the Indo-Malay region. The prevalence of these parasitic groups in zebra doves is unknown. This study estimated the prevalence and associated human risk factors of gastrointestinal nematode infections in zebra dove farming. Materials and Methods: A cross-sectional survey was conducted from January to April 2021. The study was conducted on 184 doves in three zebra dove farms. Fecal samples were collected from pooled zebra dove droppings. Major proportions and infection intensity of gastrointestinal nematodes were morphologically identified and morphometrically investigated. Associated human factors were assessed through the interview surveys among farmers. Results: Results showed that 36.96% of the zebra doves were infected. The primary nematodes were Ascaridia galli (34.78%), Heterakis gallinarum (6.52%), Trichostrongylus tenuis (2.17%), Syngamus spp. (4.35%), and Amidostomum spp. (2.17%). The primary human factors that contribute to parasitic infection were poor hygiene, food contamination with parasites, and inappropriate deworming. Conclusion: There was a high prevalence of gastrointestinal nematodes in the zebra dove in the close cage system. Human factors played key roles as risk factors, and improves farming management will help reduce parasitic infections. However, these nematodes may contribute to poor health status and poor productivity of zebra doves. Further extensive studies on clinical signs and pathological changes should be conducted.
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Ascaridia galli (Nematoda: Ascaridiidae) is the most common intestinal roundworm of chickens and other birds with a worldwide distribution. Although A. galli has been extensively studied, knowledge of the genetic variation of this parasite in detail is still insufficient. The present study examined genetic variation in the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene among A. galli isolates (n = 26) from domestic chickens in Hunan Province, China. A portion of the cox1 (pcox1) gene was amplified by polymerase chain reaction separately from adult A. galli individuals and the amplicons were subjected to sequencing from both directions. The length of the sequences of pcox1 is 441 bp. Although the intra-specific sequence variation within A. galli is 0-7.7%, the inter-specific sequence differences among other members of the infraorder Ascaridomorpha were 11.4-18.9%. Phylogenetic analyses based on the maximum likelihood method using the sequences of pcox1 confirmed that all of the Ascaridia isolates were A. galli, and also resolved three distinct clades. Taken together, the findings suggest that A. galli may represent a complex of cryptic species. Our results provide an additional genetic marker for the management of A. galli in chickens and other birds.
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Ascaridia , Genes Mitocondriais , Animais , Ascaridia/genética , Galinhas , Variação Genética , Filogenia , Análise de Sequência de DNARESUMO
This study was to assess the anthelminthic potential of Sterospermum kunthianum leaf extract against Ascaridia galli in experimentally infected broiler chickens. The extract and fractions were evaluated for in vitro inhibition and in vivo anthelmintic effects. Acute toxicity studies of extract revealed no sign of toxicity or death in birds at oral dose range of 1000-5000 and was considered safe. There was a concentration dependent decrease on inhibition of A. galli egg embryonation and deparasitization. At 100 mg/ml, albendazole (ALB) caused the highest inhibition of embryonation (195.3 ± 0.9) which was not significantly different from the decrease caused by crude methanol extract (CME) (188 ± 0.9), hexane fraction (HF) (177 ± 1.2) or ethyl acetate fraction (EAF) (168.3 ± 0.9). The highest inhibition rates (%) were 97, 94, 88 and 85 for ALB, CME, HF and EAF, respectively. The deparasitization obtained at day 21 in ALB (95.62%) treated birds was not significantly (P > 0.05) different from the 81.27% and 89.24% obtained from the crude methanol treated birds. The deparasitization caused by CME at 400 mg/kg (89.24%) was significantly higher than the one caused by EAF at the same dose (50.19%). Day 21 post treatment, significantly (P < 0.05) higher deparasitization was recorded for CME and HF at dosage of 400 mg/kg when compared to 200 mg/kg. Histopathology findings revealed necrosis of the mucosal gland and villi in chickens. In conclusion, the leaf extract and fractions S. kunthianum have been shown to possess anthelmintic activity.
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Rural chicken production in Ghana is predominantly done under the extensive system that exposes birds to parasitic infections. We investigated the prevalence of Ascaridia spp. and Heterakis spp. and as a preliminary study characterized the genetic variance of the Ascaridia galli isolates from rural chicken in Kumbungu, Savelugu and Tolon Districts in the Northern Region, Ghana. A total of 86 chickens aged 6-10 weeks were dissected and GIT inspected for nematodes. Nematode were described based on morphological features to be A. galli and H. gallinarum. Additionally, the mitochondrial cox1 gene (475 bp) of Ascaridia isolates was amplified and sequenced. The overall prevalence of nematodes was 47.67%: A. galli 37.21% and H. gallinarum 20.93%. Prevalence values of A. galli in the Kumbungu, Savelugu and Tolon Districts were 25.00%, 36.00%, 56.00%, respectively, and that of H. gallinarum, respectively were 16.67%, 28.00% and 20.00%. A Chi-square test (x2 = 6.0907, p < 0.048) showed an association of A. galli prevalence to the district of origin of birds. From 20 A. galli cox1 sequences analyzed, all sequences were identified as A. galli. Two haplotypes were recorded, namely, GHA1 and GHA2. Haplotype GHA1 was found to have wide distribution globally, whereas GHA2 appear to be novel in the present study. The data shows the importance of A. galli and H. gallinarum infection in rural chicken in northern Ghana and pave way for further epidemiological study of avian nematodes.
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Ascaridídios , Nematoides , Doenças das Aves Domésticas , Animais , Ascaridia/genética , Galinhas/parasitologia , Gana/epidemiologia , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/parasitologia , PrevalênciaRESUMO
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.
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Ascaridíase , Doenças das Aves Domésticas , Animais , Ascaridia , Ascaridíase/tratamento farmacológico , Ascaridíase/veterinária , Galinhas , Terapia de Imunossupressão/veterinária , Masculino , Óvulo , Contagem de Ovos de Parasitas/veterinária , Doenças das Aves Domésticas/tratamento farmacológicoRESUMO
One major problem of chicken (Gallus gallus domesticus) farming was various parasitic infections, especially Ascaridia galli that can cause the Ascaridiosis and is commonly found worldwide. The purpose of this study was to investigate the epidemiological situation of gastrointestinal tract parasitic infections and to develop species-specific primer for A. galli detection. A total of 247 chicken gastrointestinal tract specimens from 5 fresh markets in Bangkok. The species-specific primers of A. galli were manually designed using the mitochondrial genome at the NADH dehydrogenase subunit 4 (MT-ND 4) gene. As a result, PCR assays were optimized for the specific PCR product approximately 198 bp with the optimal temperature of 51 °C. In addition, sensitivity tests provided the detection of adult and egg stages at the minimum concentrations of 156.3 ng and 2.8 ng (70 eggs), respectively. This research can be used as preliminary information regarding the epidemic situation of gastrointestinal tract infections in chickens and detection of A. galli infection in definitive hosts, which plans programs for the effective control and prevention of parasitic infections.
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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.
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Ascaridíase , Doenças das Aves Domésticas , Animais , Ascaridia/genética , Ascaridíase/diagnóstico , Ascaridíase/veterinária , Bangladesh , Galinhas , Óvulo , Filogenia , Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/diagnósticoRESUMO
1. Ascarids (Ascaridia galli and Heterakis spp.) are highly prevalent in free-range laying hens. Ascarid eggs survive for long periods in soil, and one preventive measure is to add litter material to areas close to the henhouse. In this study, recovery rates of ascarid eggs from three common litter materials, namely pea gravel, beech (Fagus sylvatica) and spruce (Picea abies) wood chips were compared to recovery rates from soil. 2. Materials were mixed with faeces containing 1,408 ascarid eggs per g of faeces, placed in plastic fruit boxes and exposed to natural weather conditions in a randomised block design with six replicates per treatment. 3. Numbers of ascarid eggs were quantified at 28 time points over 3.5 years. Ascarid eggs were recovered for over three years from all materials and completely disappeared during the fourth winter of exposure. Time needed to get to a 50% reduction in ascarid eggs did not differ between litter materials and soil (242 to 269 days). A 99% reduction was reached significantly (P < 0.001) earlier in pea gravel (548 days) than in the three other materials, and earlier in the two wood chips (day 682 for beech, day 692 for spruce, P < 0.05) than in soil (1,277 days). 4. Accumulation of ascarid eggs in the area close to the henhouse can be reduced by any of the tested litter materials compared to bare soil. Adding litter to this area is highly recommended for free-range layer farmers in order to reduce numbers of infective ascarid eggs.