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Coccidiosis, caused by Eimeria parasites, is a major threat to global poultry production, and increasing restrictions on conventional anticoccidial drugs highlight the need for safer, more sustainable alternatives. Progress has been hindered by the lack of rapid, sensitive, and animal-sparing in vitro assays for quantifying parasite replication and drug efficacy. This study reports the development of a novel bioluminescent platform for anticoccidial screening based on a genetically modified Eimeria tenella line expressing NanoLuc luciferase (EtNluc). Parasite-associated bioluminescence enabled rapid and quantitative monitoring of intracellular development, allowing the tracking of different replication phases through schizont formation and merozoite release. Time course analysis showed minimal changes in relative light units (RLU) between 2 and 24 hours post infection (hpi), followed by a marked increase between 24 and 72 hpi, consistent with parasite replication. Among the tested multiplicities of infection (MOI), 4:1 exhibited the fastest growth, described by a linear model (slope = 2908 RLU/h, R 2 = 0.84). A same-well repeated-measure analysis (2 and 72 hpi) confirmed the dose-dependent replication, with mean slopes of 2052.85, 765.07 and 523.63 RLU/h, respectively, supporting the selection of the MOI 4:1 for anticoccidial screening. These experimental conditions were used to evaluate the anticoccidial efficacy of commercial anticoccidial drugs (salinomycin and robenidine) and natural compounds (thyme and oregano essential oils, thymol, and carvacrol) under two experimental designs: short pre-incubation of sporozoites, and continuous exposure throughout intracellular development. Pre-incubation with commercial anticoccidials reduced invasion approximately to 65% for salinomycin and 44% for robenidine, whereas the essential oils and their bioactive constituents inhibited invasion by 30-55%, and reduced the replication slope to 33-60% of control values. Continuous exposure significantly impaired intracellular development for all treatments, reducing replication to 10-30% of controls, providing additional evidence that plant-derived compounds can complement commercial anticoccidials for integrated strategy for coccidiosis control in chickens. Overall, the EtNluc bioluminescent system provided a rapid, sensitive, and scalable method for quantifying E. tenella growth, suitable for in vitro anticoccidial screening, supporting the characterization of novel anticoccidial while reducing reliance on animal experimentation.

The poultry industry is one of the main providers of protein for the world's population, but it faces great challenges including coccidiosis, one of the diseases with the most impact on productive performance. Coccidiosis is caused by protozoan parasites of the genus Eimeria , which are a group of monoxenous obligate intracellular parasites. Seven species of this genus can affect chickens ( Gallus gallus) , each with different pathogenic characteristics and targeting a specific intestinal location. Eimeria alters the function of the intestinal tract, generating deficiencies in the absorption of nutrients and lowering productive performance, leading to economic losses. The objective of this manuscript is to review basic concepts of coccidiosis, the different Eimeria species that infect chickens, their life cycle, and the most sustainable and holistic methods available to control the disease.

Coccidiosis is a deadly disease that hampers chicken’s productivity and welfare. Thus, the disease is a major menace to the global poultry industry. Coccidiosis which is caused by the apicomplexan parasite of the genus Eimeria has seven known species which affect the different parts of the intestinal tract of chickens. The disease which occurs by ingestion of sporulated oocyst has been associated with poor poultry management system. Mixed infection among the species of this parasite contributes to both pathogenicity and misdiagnosis of the disease. A progress in identification and diagnosis approach which cuts across pathological, morphological and molecular has been reported for this parasite. Control measures which include anticoccidial drugs, vaccines and natural products have dominated literature for this disease. However, the emergence of genetic and antigenic diversity with implication on resistance to anticoccidials among different strains of Eimeria parasite has generated concerns on the effectiveness of the current anticoccidial vaccines. A new look on the control strategy therefore becomes imperative. This study reviews the current trends on the identification and control of chicken coccidiosis with focus on (1) Avian coccidiosis (2) Epidemiology of chicken coccidiosis (3) Eimeria parasite and distribution in poultry (4) Diagnosis of Eimeria parasite (5) Control measures of coccidiosis (6) Threats posed by genetic and antigenic diversity of Eimeria parasite on coccidiosis control. Genomic study on diversity of Eimeria parasite becomes imperative for effective vaccine design against coccidiosis.

Anticoccidial drug resistance in coccidia has been known since the 1960s, and experimental selection for resistance in the laboratory has been done for most available products. However, routine testing for sensitivity in field isolates has only begun in recent years. Poultry producers are faced with greater challenges because of consumer preferences for ‘antibiotic-free’ products and a lack of new products for disease control. The classification of some of our most effective anticoccidials as ‘antibiotics’ has severely limited their use, causing us to rely on older products that are prone to resistance development. The interaction between coccidiosis and other diseases such as necrotic enteritis places more importance on routine testing for drug sensitivity. In this review, we have summarized the use of the anticoccidial sensitivity test (AST) as it is conducted and interpreted by poultry health specialists.

This study aimed to assess the efficacy of an anticoccidial vaccine and the anticoccidial activity of Aloe vera in broiler chickens infected with Eimeria tenella (E. tenella). A total of 225 healthy, sexless, one-day-old broiler chicks (avian48) from a commercial broiler company were randomized into nine experimental groups of 25 chicks. The groups were as follows: Group 1 (control, vaccinated, non-infected), Group 2 (vaccinated and infected with 5 × 104 sporulated oocysts), Group 3 (vaccinated, infected with 5 × 104 sporulated oocysts, and treated with Aloe vera), Group 4 (infected with 5 × 104 sporulated oocysts and treated with Aloe vera), Group 5 (positive control, infected with 5 × 104 sporulated oocysts), Group 6 (challenged with 5 × 104 sporulated oocysts and then treated with amprolium), Group 7 (treated with amprolium), Group 8 (blank control negative group), and Group 9 (treated with Aloe vera gel).Various parameters were evaluated, including clinical signs, growth performance, oocyst shedding, hematological and immunological parameters, and pathological lesion scoring. The results demonstrated that Aloe vera improved growth performance, reduced oocyst shedding, and decreased caecal lesion scores in E. Tenella-infected broiler chicks. The use of Aloe vera in combination with either amprolium or anticoccidial vaccines provided a potential solution to the issues of drug resistance and drug residues.In conclusion, this study provides valuable insights regarding the control of coccidiosis in broilers. Supplementing the chicken diet with Aloe vera had beneficial effects on the pathogenicity and infectivity of E. tenella, making it a cost-effective alternative as an herbal extract with no adverse side effects for coccidiosis control. These findings suggest that Aloe vera can be considered a potential candidate for inclusion in broiler diets for effective coccidiosis control.Research HighlightsEfficacy of an anticoccidial vaccine, and Aloe vera extracts in broiler chickens infected with E. tenella.Supplementation of Aloe vera had improved growth performance, reduced oocyst shedding, and decreased caecal lesion scores in infected broiler chicks.The combination of Aloe vera and anticoccidial vaccines showed promise for addressing drug resistance and residues.The clinical signs, growth performance, oocyst shedding, haematological and immunological parameters, and pathological lesion scoring were all evaluated.Aloe vera is a safe and cost-effective alternative to herbal extracts for the control of coccidiosis in broilers.The findings suggest incorporating Aloe vera into broiler diets for effective coccidiosis control.

Avian species have long struggled with the problem of coccidiosis, a disease that affects various parts of the intestine, including the anterior gut, midgut, and hindgut. Among different types of coccidiosis, cecal coccidiosis is particularly dangerous to avian species. Chickens and turkeys are commercial flocks; thus, their parasites have remained critical due to their economic importance. High rates of mortality and morbidity are observed in both chickens and turkeys due to cecal coccidiosis. Coccidiostats and coccidiocidal chemicals have traditionally been added to feed and water to control coccidiosis. However, after the EU banned their use because of issues of resistance and public health, alternative methods are being explored. Vaccines are also being used, but their efficacy and cost-effectiveness remain as challenges. Researchers are attempting to find alternatives, and among the alternatives, botanicals are a promising choice. Botanicals contain multiple active compounds such as phenolics, saponins, terpenes, sulfur compounds, etc., which can kill sporozoites and oocysts and stop the replication of Eimeria . These botanicals are primarily used as anticoccidials due to their antioxidant and immunomodulatory activities. Because of the medicinal properties of botanicals, some commercial products have also been developed. However, further research is needed to confirm their pharmacological effects, mechanisms of action, and methods of concentrated preparation. In this review, an attempt has been made to summarize the plants that have the potential to act as anticoccidials and to explain the mode of action of different compounds found within them.

The aims of this study were to determine the immune response and the anticoccidial activity induced by and leaves in rabbits infected with and . Thirty-five-day-old rabbits, free from coccidia, were infested with 2.10 oocysts of and , then received the acetone extract of the leaves of and at different doses by oral gavage. The inhibition of the excretion of oocysts was evaluated by the McMaster technique and the levels of cytokines (IL-4 and IL-12) and immunoglobulin IgG were assayed by the ELISA method. The efficacy on and oocysts was 95.43% and 96.53% for and at 1000 mg/kg bw against 98% for the positive control. Interestingly the plant extracts increased the production of interleukin (IL) and immunoglobulins (Ig) compared to controls. Plasma IL-4 levels (pg/ml) in rabbits were 128.94 and 131.38; those of IL-12 (pg/ml) were 395.55 and 426.56, and then for those of IgG (μg/ml) were 14.70 and 13.94 respectively with the acetone extracts of and on D14 PT at 1000 mg/kg bw. This study indicates that and can be used as an alternative to synthetic anticoccidials. These plants could be used to increase the resistance of the immune system of rabbits to infestations of Eimeria species in rabbit farms.

Piglet coccidiosis due to Cystoisospora suis is a major cause of diarrhea and poor growth worldwide. It can effectively be controlled by application of toltrazuril (TZ), and oral formulations have been licensed for many years. Recently, the first parenteral formulation containing TZ in combination with iron (gleptoferron) was registered in the EU for the prevention of coccidiosis and iron deficiency anemia, conditions in suckling piglets requiring routine preventive measures. This study evaluated the absorption and distribution of TZ and its main metabolite, toltrazuril sulfone (TZ-SO2), in blood and intestinal tissues after single oral (20 mg/kg) or single intramuscular (45 mg/piglet) application of TZ. Fifty-six piglets were randomly allocated to the two treatment groups. Animals were sacrificed 1-, 5-, 13-, and 24-days post-treatment and TZ and TZ-SO2 levels were determined in blood, jejunal tissue, ileal tissue, and mixed jejunal and ileal content (IC) by high performance liquid chromatography (HPLC). Intramuscular application resulted in significantly higher and more sustained concentrations of both compounds in plasma, intestinal tissue, and IC. Higher concentrations after oral dosing were only observed one day after application of TZ in jejunum and IC. Toltrazuril was quickly metabolized to TZ-SO2 with maximum concentrations on day 13 for both applications. Remarkably, TZ and TZ-SO2 accumulated in the jejunum, the primary predilection site of C. suis, independently of the administration route, which is key to their antiparasitic effect.

Coccidiosis remains one of the major problems of the poultry industry. Caused by Eimeria species, Coccidiosis is a contagious parasitic disease affecting poultry with great economic significance. Currently, in order to prevent health problems caused by this disease, broiler farmers make extensive use of coccidiostats in poultry feed, maintaining animal health and, in some cases, enhancing feed conversion. The presence of unauthorized substances, residues of veterinary products and chemical contaminants in the food industry is of concern, since they may pose a risk to public health. As the use of coccidiostats has been increasing without any requirements for veterinary prescription, research and surveillance of coccidiostat residues in poultry meat is becoming imperative. This review presents an up-to-date comprehensive discussion of the state of the art regarding coccidiosis, the most used anticoccidials in poultry production, their mode of action, their prophylactic use, occurrence and the European Union (EU) applicable legislation.

Increase in drug resistance as well as ineffective immunization efforts against various pathogens (viruses, bacteria and fungi) pose a significant threat to the poultry industry. Spirulina is one of the most widely used natural ingredients which is becoming popular as a nutritional supplement in humans, animals, poultry and aquaculture. It contains protein, vitamins, minerals, fatty acids, pigments, and essential amino acids. Moreover, it also has considerable quantities of unique natural antioxidants including polyphenols, carotenoids, and phycocyanin. Dietary supplementation of Spirulina can beneficially affect gut microbial population, serum biochemical parameters, and growth performance of chicken. Additionally, it contains polyphenolic contents having antibacterial effects. Spirulina extracts might inhibit bacterial motility, invasion, biofilm formation, and quorum sensing in addition to acting directly on the bacterium by weakening and making the bacterial cell walls more porous, subsequently resulting in cytoplasmic content leakage. Additionally, Spirulina has shown antiviral activities against certain common human or animal viruses and this capability can be considered to exhibit potential benefits against avian viruses also. Spirulan, a calcium-rich internal polysaccharide of Spirulina, is potentially responsible for its antiviral effect through inhibiting the entry of several viruses into the host cells, boosting the production of nitric oxide in macrophages, and stimulating the generation of cytokines. Comparatively a greater emphasis has been given to the immune modulatory effects of Spirulina as a feed additive in chicken which might boost disease resistance and improve survival and growth rates, particularly under stress conditions. This manuscript reviews biological activities and immune-stimulating properties of Spirulina and its potential use as a dietary supplement in poultry to enhance growth, gut health and disease resistance.