Evaluation of semi-quantitative colorimetric assays based on loop-mediated isothermal amplification indicators by using image analysis.

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.

Hepatic transcriptomic analysis reveals differential regulation of metabolic and immune pathways in three strains of chickens with distinct growth rates exposed to mixed parasite infections.

During parasite infections, the liver may prioritise immune-related pathways over its metabolic functions. Intestinal infections caused by Ascaridia galli and Heterakis gallinarum impair feed intake, nutrient absorption, and weight gain. Histomonas meleagridis, vectored by H. gallinarum, can also damage liver tissues, potentially impairing liver functions. This study examined the hepatic gene expression in three strains of chickens: Ross-308 (R), Lohmann Brown Plus (LB), and Lohmann Dual (LD), 2 weeks after an experimental infection (n = 18) with both A. galli and H. gallinarum or kept as uninfected control (n = 12). Furthermore, H. gallinarum infection led to a co-infection with H. meleagridis. The mixed infections reduced feed intake and the average daily weight gain (P < 0.001). The infections also increased the plasma concentrations of alpha (1)-acid glycoprotein and the antibody titre against H. meleagridis (P = 0.049), with no strain differences (P > 0.05). For host molecular response, 1887 genes were differentially expressed in LD, while 275 and 25 genes were differentially expressed in R and LB, respectively. The up-regulated genes in R and LD were mostly related to inflammatory and adaptive immune responses, while down-regulated genes in LD were involved in metabolic pathways, including gluconeogenesis. Despite performance differences among the strains, worm burdens were similar, but hepatic molecular responses differed significantly. Moreover, there was an indication of a shift in hepatic functions towards immune-related pathways. We, therefore, conclude that the liver shifts its functions from metabolic to immune-related activities in chickens when challenged with mixed parasite species.

An automated faecal egg count system for detection of Ascaridia galli ova in chickens.

Chicken production has increased over the past decade, resulting in a concomitant rise in the demand for more humane options for poultry products including cage-free, free-range, and organic meat and eggs. These husbandry changes, however, have come hand-in-hand with increased prevalence of Ascaridia galli infection, which can cause clinical disease in chickens as well as the occasional appearance of worms in eggs. Additionally, development of anthelmintic resistance in closely related helminths of turkeys highlights the need for closely monitored anthelmintic treatment programs. Manual faecal egg counts (FECs) can be time-consuming and require specialist training. As such, this study sought to validate an automated FEC system for use in detection and quantification of A. galli eggs in chicken faeces. Automated counts using the Parasight System (PS) were compared to traditional manual McMaster counting for both precision and correlation between methods. Overall, ten repeated counts were performed on twenty individual samples for a total of 200 counts performed for each method. A strong, statistically significant correlation was found between methods (R2 = 0.7879, P < 0.0001), and PS counted more eggs and performed with statistically significant higher precision (P = 0.0391) than manual McMaster counting. This study suggests that PS is a good alternative method for performing A. galli FECs and provides a new tool for use in helminth treatment and control programs in chicken operations.

Feasibility of a DNA biosensor assay based on loop-mediated isothermal amplification combined with a lateral flow dipstick assay for the visual detection of Ascaridia galli eggs in faecal samples.

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 70 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.

Sequence analyses of mitochondrial gene may support the existence of cryptic species within Ascaridia galli.

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.

Ascarid eggs disappear faster from gravel and wood chips than from soil.

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.

Survival and development of chicken ascarid eggs in temperate pastures.

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

Evaluation of benzimidazole resistance status in Ascaridia galli.

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

Demonstration of biological activities of extracts from Isodon rugosus Wall. Ex Benth: Separation and identification of bioactive phytoconstituents by GC-MS analysis in the ethyl acetate extract.

BACKGROUND: Since long, natural sources have been explored for possible managements of various diseases. In this context, the study is designed to evaluate Isodon rugosus Wall. ex Benth for biological potentials including antibacterial, anthelmintic, insecticidal, anti-termites and anti-Pharaoh activities followed by GC-MS analysis of active fraction to identify various bioactive compounds. METHODS: I. rugosus was investigated against eight bacterial strains using well diffusion method and microdilution method with ceftriaxone as positive control. Similarly, the insecticidal activity was carried out against Tribolium castaneum, Rhyzopertha dominica, Monomorium pharaonis and Heterotermis indicola following contact toxicity method. Likewise, anthelmintic activity was performed against Ascaridia galli and Pherethima posthuma using albendazole as positive control, in which the paralysis and death times of the worms were observed. The GC-MS analysis of the most active solvent fraction was performed for identifications of various bioactive compounds. RESULTS: Among the tested samples of I. rugosus, flavonoids and ethyl acetate fraction exhibited high antibacterial activities. The crude saponins showed highest anthelmintic activity against Pherethima posthuma and Ascaridia galli with death times of 27.67 and 29.22 min respectively at concentrations of 40 mg/ml. In insecticidal activity, chloroform fraction and saponins exhibited notable results against R. dominica (60 and 70%) and T. castaneum (70 and 76%) at concentration of 200 mg/ml. In anti-termite assay, all the plant samples showed overwhelming results, i.e. all the 25 termites were killed on the 3rd day. Similarly, in anti-Pharaoh activity, the chloroform, ethyl acetate and saponins fractions were most potent, each exhibiting LD50 of <0.1 mg/ml. In GC-MS analysis, total of 57 compounds were identified. Some of the bioactive compounds identified in GC-MS analysis are palmitic acid, hinokiol, α-amyrin, phytol, ethyl linolate, cyclohexanone, hinokione, methyl palmitate, ethyl palmitate and stigmasterol acetate. CONCLUSIONS: Based on our current results, it can be concluded that I. rugosus possess strong antibacterial, insecticidal and anthelmintic potentials having crude saponins and ethyl acetate as the most active fractions. The GC-MS analysis and biological assays reveal that ethyl acetate fraction is a suitable target for the isolation of diverse array of bioactive compounds.

Evaluation of anthelmintic efficacy of ethanolic leaf extract of Juglans regia L. on Ascaridia galli: a comprehensive in vitro and in vivo study.

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.

Periodicity of Ascaridia galli egg excretion in experimentally infected chicken in the Philippines.

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.

Assay for the simultaneous detection of Raillietina spp. (R. echinobothrida, R. tetragona, and R. cesticillus) and Ascaridia galli infection in chickens using duplex loop-mediated isothermal amplification integrated with a lateral flow dipstick assay.

Raillietina species and Ascaridia galli are two of the significant intestinal parasites that affect chickens in a free-range system production. They destroy the intestinal mucosa layer, leading to several clinical symptoms such as weight loss, a slowed growth rate, and economic value loss. Thus, the objective of this study was to develop an assay for simultaneously detecting Raillietina spp. (R. echinobothrida, R. tetragona, and R. cesticillus) and A. galli in a single reaction using duplex loop-mediated isothermal amplification (dLAMP) coupled with a lateral flow dipstick (LFD) assay. The analytical specificity of the dLAMP-LFD assay showed a high specific amplification of Raillietina spp. and A. galli without non-target amplification. Regarding the analytical sensitivity, this approach was capable of simultaneously detecting concentrations as low as 5 pg/µL of mixed-targets. To evaluate the efficiency of the dLAMP assay, 30 faecal samples of chickens were verified and compared through microscopic examination. The dLAMP-LFD assay and microscopic examination results showed kappa values of Raillietina spp. and A. galli with moderate (K= 0.615) to high (K= 1) agreements, respectively, while the McNemar's test indicated that the efficiency between assays was not significantly different. Therefore, the developed dLAMP-LFD assay can be used as an alternative screening method to the existing classical method for epidemiological investigation, epidemic control, and farm management, as well as for addressing poultry health problems.

The complete mitochondrial genome of the chicken roundworm Ascaridia galli (Nematoda: Ascaridiidae).

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.

Comparative Effect of Ascaridia Galli Infection on the Two Laying Hen Lines on the Liver Function and Immune Response.

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.

In vitro evaluation of the effects of methanolic plant extracts on the embryonation rate of Ascaridia galli eggs.

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.

Interactions between the helminth and intestinal microbiome in smallholder chicken farming systems.

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.

Ascaridia galli infection in chicken: Pathobiology and immunological orchestra.

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.

Ascaridia galli challenge model for worm propagation in young chickens with or without immunosuppression.

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.

In vitro and in vivo anthelmintic effects of Sterospermum kunthianum (Cham-Holl) leaf extract against Ascaridia galli in experimentally infected broiler chickens.

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.

First report of gastrointestinal nematodes and coccidia parasites from free-range chickens in Mafeteng district, Lesotho.

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.