Metabolic Blockade-Based Genome Mining of Sea Anemone-Associated Streptomyces sp. S1502 Identifies Atypical Angucyclines WS-5995 A-E: Isolation, Identification, Biosynthetic Investigation, and Bioactivities.

Marine symbiotic and epiphyte microorganisms are sources of bioactive or structurally novel natural products. Metabolic blockade-based genome mining has been proven to be an effective strategy to accelerate the discovery of natural products from both terrestrial and marine microorganisms. Here, the metabolic blockade-based genome mining strategy was applied to the discovery of other metabolites in a sea anemone-associated Streptomyces sp. S1502. We constructed a mutant Streptomyces sp. S1502/Δstp1 that switched to producing the atypical angucyclines WS-5995 A-E, among which WS-5995 E is a new compound. A biosynthetic gene cluster (wsm) of the angucyclines was identified through gene knock-out and heterologous expression studies. The biosynthetic pathways of WS-5995 A-E were proposed, the roles of some tailoring and regulatory genes were investigated, and the biological activities of WS-5995 A-E were evaluated. WS-5995 A has significant anti-Eimeria tenell activity with an IC50 value of 2.21 µM. The production of antibacterial streptopyrroles and anticoccidial WS-5995 A-E may play a protective role in the mutual relationship between Streptomyces sp. S1502 and its host.

Synthesis and evaluation of febrifugine derivatives as anticoccidial agents.

A series of new febrifugine derivatives with a 4(3H)-quinazolinone scaffold were synthesized and evaluated for their anticoccidial activity both in vitro and in vivo. The targets' in vitro activity against Eimeria tenella was studied using quantitative real-time reverse transcription polymerase chain reaction and Madin-Darby bovine kidney cells. Most of these compounds demonstrated anticoccidial efficacy, with inhibition ratios ranging from 3.3% to 85.7%. Specifically, compounds 33 and 34 showed significant inhibitory effects on the proliferation of E. tenella and exhibited lower cytotoxicity compared to febrifugine. The IC50 values of compounds 33 and 34 were 3.48 and 1.79 µM, respectively, while the CC50 values were >100 µM for both compounds. Furthermore, in a study involving 14-day-old chickens infected with 5 × 104 sporulated oocysts, treatment with five selected compounds (22, 24, 28, 33, and 34), which exhibited in vitro inhibition rate of over 50% at 100 µM, at a dose of 40 mg/kg in daily feed for 8 consecutive days showed that compound 34 possessed moderate in vivo activity against coccidiosis, with an anticoccidial index of 164. Structure-activity relationship studies suggested that spirocyclic piperidine may be a preferable substructure to maintain high effectiveness in inhibiting Eimeria spp., when the side chain 1-(3-hydroxypiperidin-2-yl)propan-2-one was replaced.

Synergistic approach for treatment of chicken coccidiosis using berberine--A plant natural product.

Despite the advent of anticoccidial drugs and vaccines, coccidiosis continues to result in substantial economic losses to the poultry industry. Berberine, a natural alkaloid is well known in studies involving synergistic approaches, thereby reducing the dosage of principal drugs. Therefore, a study was designed to see whether a synergistic anticoccidial effect could be obtained between amprolium and berberine, in vivo using broiler chicken. Anticoccidial activity was measured in comparison to the reference drug amprolium on the basis of oocyst output reduction, mean weight gain and feed conversion ratio. Oocyst output was measured using Mc-Masters counting technique. Different combinations of berberine and amprolium were tested and out of which 1:1 ratio was the most effective for controlling these parasites. Oral gavaging of 100(50 + 50) mg/kg body weight of 1:1 ratio of amprolium and berberine caused the equivalent reduction in number of oocysts (38.85 ± 9.61) one day prior to that of standard drug amprolium (49.95 ± 16.65) as well as pure berberine (44.4 ± 9.61) used in the study. Weight gain of birds was also highest in the synergistic group (1547.43 ± 12.86) among all the infected groups. Besides feed conversion ratio in the synergistic group was also better (1.387 ± 0.026). The results of this study proved the effectiveness of both amprolium and berberine and revealed synergism between amprolium and berberine against coccidian oocysts, confirmed by significant reduction in the number of coccidian oocysts shed in the feces, leading to better weight gain and improved feed conversion ratio. The study deep-rooted the synergistic potential of berberine, a natural bioactive compound for controlling a protozoan parasite and the results of this study corroborate with its use for treatment of severe diarrhoea, amoebiasis and intestinal infections.

Synthesis and evaluation of the anticoccidial activity of trifluoropyrido[1,2-a]pyrimidin-2-one derivatives.

Screening of our chemical library to discover new molecules exhibiting in vitro activity against the invasion of host cells by Eimeria tenella revealed a lead compound with an IC50 of 15µM. Structure-activity relationship studies were conducted with 34 newly synthesized compounds to identify more active molecules and enhance in vitro activity against the parasite. Four compounds were more effective in inhibiting MDBK cell invasion in vitro than the lead compound.

Effects of functional oils on coccidiosis and apparent metabolizable energy in broiler chickens.

The objective of the present study was to investigate the effects of a mixture of functional oils (Essential, Oligo Basics Agroind. Ltda) on performance response of chickens challenged with coccidiosis and the determination of apparent metabolizable energy (AME), nitrogen-corrected apparent metabolizable energy (AMEn), the coefficients of protein and ether extract digestibility and intestinal morphology of broilers fed with diets containing Essential. In Exp. 1, a completely randomized design (CRD) was used, with one control diet without Essential inclusion with coccidiosis (Eimeria acervulina, Eimeria maxima, and Eimeria tenella) challenged birds and two different inclusion rates of Essential (1.5 kg/ton and 2 kg/ton) with coccidiosis-challenged and non-challenged birds for each inclusion rate, using 10 replicates and 50 birds per experimental unit. After 7 d of coccidiosis challenge, the livability was approximately 10% lower (p<0.05) for the control group. Intestinal lesion scores were lower (p<0.05) in the anterior intestine and the cecum for the chickens supplemented. Feed efficiency and growth rate were improved in birds supplemented with Essential (p<0.05) before the coccidiosis challenge and during the first 7 d post infection. In Exp. 2, a CRD was used, with one control diet without Essential inclusion and one diet with inclusion of Essential (1.5 kg/ton), using nine replications and 33 chicks per pen. The diets with Essential yielded approximately 4% higher AME (p = 0.003) and AMEn (p = 0.001). Essential supplementation increased villus height in the jejunum on d 14 (p<0.05). Villus height:crypt depth ratio for the supplemented birds was larger (p<0.05) in the jejunum on d 7, larger (p<0.05) in the jejunum and ileum on d 14. In conclusion, these functional oils improved the energy utilization and the livability and decreased lesions caused by coccidiosis in supplemented birds.

Portulaca oleracea exhibited anti-coccidian activity, fortified the gut microbiota of Hu lambs.

Coccidia of the genus Eimeria are important pathogens that cause coccidiosis in livestock and poultry. Due to the expansion of intensive farming, coccidiosis has become more difficult to control. In addition, the continued use of anti-coccidiosis drugs has led to drug resistance and residue. Some herbs used in traditional Chinese medicine (TCM) have been shown to alleviate the clinical symptoms of coccidiosis, while enhancing immunity and growth performance (GP) of livestock and poultry. Previous in vitro and in vivo studies have reported that the TCM herb Portulaca oleracea exhibited anti-parasitic activities. In total, 36 female Hu lambs were equally divided into six treatment groups: PL (low-dose P. oleracea), PH (high-dose P. oleracea), PW (P. oleracea water extract), PE (P. oleracea ethanol extract), DIC (diclazuril), and CON (control). The treatment period was 14 days. The McMaster counting method was used to evaluate the anti-coccidiosis effects of the different treatments. Untargeted metabolomics and 16S rRNA gene sequencing were used to investigate the effects of treatment on the gut microbiota (GM) and GP. The results showed that P. oleracea ameliorated coccidiosis, improved GP, increased the abundances of beneficial bacteria, and maintained the composition of the GM, but failed to completely clear coccidian oocysts. The Firmicutes to Bacteroides ratio was significantly increased in the PH group. P. oleracea increased metabolism of tryptophan as well as some vitamins and cofactors in the GM and decreased the relative content of arginine, tryptophan, niacin, and other nutrients, thereby promoting intestinal health and enhancing GP. As an alternative to the anti-coccidiosis drug DIC, P. oleracea effectively inhibited growth of coccidia, maintained the composition of the GM, promoted intestinal health, and increased nutrient digestibility.

Anticoccidial activity of Aloe Vera Leafs' aqueous extract and vaccination against Eimeria tenella: pathological study in broilers.

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.

Effects of dietary water-soluble extract of rosemary supplementation on growth performance and intestinal health of broilers infected with Eimeria tenella.

This study was conducted to explore the effect of dietary supplementation of water-soluble extract of rosemary (WER) on growth performance and intestinal health of broilers infected with Eimeria tenella (E. tenella), and evaluate the anticoccidial activity of WER. 360 1-d-old Chinese indigenous male yellow-feathered broiler chickens were randomly allocated to six groups: blank control (BC) group and infected control (IC) group received a basal diet; positive control (PC) group, received a basal diet supplemented with 200 mg/kg diclazuril; WER100, WER200, and WER300 groups received a basal diet containing 100, 200, and 300 mg/kg WER, respectively. On day 21, all birds in the infected groups (IC, PC, WER100, WER200, and WER300) were orally gavaged with 1 mL phosphate-buffered saline (PBS) of 8 × 104 sporulated oocysts of E. tenella, and birds in the BC group were administrated an aliquot of PBS dilution. The results showed that dietary supplementation of 200 mg/kg WER increased the average daily gain of broilers compared to the IC group from days 22 to 29 (P < 0.001). The anticoccidial index values of 100, 200, and 300 mg/kg WER were 137.49, 157.41, and 144.22, respectively, which indicated that WER exhibited moderate anticoccidial activity. Compared to the IC group, the groups supplemented with WER (100, 200, and 300 mg/kg) significantly lowered fecal oocyst output (P < 0.001) and cecal coccidia oocysts, alleviated intestinal damage and maintained the integrity of intestinal epithelium. Dietary supplementation with WER significantly improved antioxidant capacity, elevated the levels of secretory immunoglobulin A, and diminished inflammation within the cecum, particularly at a dosage of 200 mg/kg. The results of this study indicated that dietary supplementation with 200 mg/kg WER could improve broiler growth performance and alleviate intestinal damage caused by coccidiosis.

Avian coccidiosis, a prevalent parasitic disease caused by Eimeria protozoa, leads to significant economic losses in the global poultry industry. Currently, the control of coccidiosis in chickens primarily relies on chemical and ionophore anticoccidials. However, the long-term use of these compounds has resulted in the development of drug-resistant strains, presenting a critical challenge. Additionally, the toxic and side effects of ionophore anticoccidials have become increasingly apparent. Thus, there is an urgent need to find economical and environmentally friendly measures to control coccidiosis in chickens. In this study, we established a model of Eimeria tenella infection in broilers to explore whether the water-soluble extract of rosemary (WER) could serve as an alternative method for controlling avian coccidiosis. Our results showed that dietary supplementation with WER (100, 200, and 300 mg/kg) had a beneficial anticoccidial effect, alleviating intestinal damage caused by coccidiosis by enhancing the intestinal antioxidant defense and activating the immune function of the infected broilers. Specifically, dietary supplementation with 200 mg/kg WER emerged as a promising strategy for controlling avian coccidiosis in the poultry industry.

The anticoccidial effect of alcoholic extract of Citrullus colocynthis fruit and Juglans regia peel| in experimentally infected domestic chicken.

One of the main illnesses within the poultry industry is coccidiosis. Anticoccidial medicines applied in the poultry industry show many shortcomings and new control measures are necessary. The current research aimed to study the effect of extract of Citrullus colocynthis and Juglans regia peel on growth performance, gut bacteria, Haematological, Anticoccidial Index (ACI), and Optimum Anticoccidial Activity (OAA) of coccidiosis-infected domestic chicken. The maximum weight gain was observed in the groups treated with 0.001% and 0.01% C. colocynthis extract. Moreover, 0.01% C. colocynthis extract treatment increased two factors of ACI and OAA by 121.42 and 109, respectively, which were higher than commercial anticoccidial (Sulfaclozine). The extract of C. colocynthis fruit and J. regia peel decreased monocytes and eosinophils haematological factors and increased basophils in birds infected with Eimeria. Both extracts modulated intestinal microbiome haematological factors in birds infected with Eimeria, while J. regia peel extract had better performance than C. colocynthis fruit extract. These results indicate that used C. colocynthis and J. regia extracts have an anti-coccidial effect and the potential to control Eimeria infection.

Bioactive Compounds and In Vitro Antioxidant and Anticoccidial Activities of Opuntia ficus-indica Flower Extracts.

The objective of the present study is to identify the biochemical compounds extracted from OFI flowers using ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry and to evaluate their in vitro antioxidant activities and anticoccidial effects on the destruction of Eimeria oocysts isolated from naturally infected chickens. A domestic microwave was used with a refrigerant to condense the vapors generated during the extraction. The flavonoid and phenolic compound contents of the OFI flowers were determined according to standard methods. DPPH radical and H2O2 scavenging capacities were used to assess the antioxidant activity. Regarding the anticoccidial activity, the Eimeria spp. oocysts used were isolated from the fresh feces of infected broilers and were determined in triplicate by incubation at an ambient temperature for 24 h. The results highlighted the considerable influence of the optimized acetone concentration, ratio, irradiation time, and microwave power parameters on the phenolic content and antioxidant activities. Our results revealed significant matches between the predicted and experimental values of the models. Molecular analysis revealed the presence of several biophenol classes such as quercetin, isorhamnetin 3-O-rutinoside, and quercetin-3-O-rutinoside. OFI flower extracts inhibited sporulation and damaged the morphology of Eimeria oocysts compared with normal sporulated Eimeria oocysts containing sporocysts. In conclusion, the optimized conditions were validated and found to fit very well with the experimental values. These findings suggest that the flowers of OFI should be considered sources of antioxidants. The results of the present study revealed that OFI flower extracts have anticoccidial activities against Eimeria-spp.-induced infection in broiler chickens.

In vivo anticoccidial activity and immune response of Moringa oleifera and Vernonia amygdalina leaves against Eimeria magna and Eimeria media in rabbits.

Introduction: The aims of this study were to determine the immune response and the anticoccidial activity induced by Moringa oleifera and Vernonia amygdalina leaves in rabbits infected with Eimeria magna and Eimeria media. Methods: Thirty-five-day-old rabbits, free from coccidia, were infested with 2.103 oocysts of Eimeria magna and Eimeria media, then received the acetone extract of the leaves of Moringa oleifera and Vernonia amygdalina at different doses by oral gavage. Results and discussion: 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 in vivo efficacy on E. magna and E. media oocysts was 95.43% and 96.53% for Moringa oleifera and Vernonia amygdalina 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 Moringa oleifera and Vernonia amygdalina on D14 PT at 1000 mg/kg bw. This study indicates that Moringa oleifera and Vernonia amygdalina 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.

Green Synthesis of ZnO/CuO Nanocomposites Using Parsley Extract for Potential In Vitro Anticoccidial Application.

In this study, a simple, low-cost, and environmentally friendly method for the green synthesis of ZnO/CuO nanocomposites (NCs) using parsley extract was developed. The phytochemical components in the parsley leaf extract reacted with precursor salts in solution and yielded ZnO/CuO NCs. The synthesis of the green-synthesized NCs was confirmed via various characterization techniques, including UV-vis spectroscopy, X-ray diffraction (XRD) analysis, energy-dispersive X-ray (EDX), transmission electron microscopy (TEM), and field emission scanning electron microscopy (FE-SEM). Subsequently, the NCs were subjected to rigorous in vitro evaluation of their anticoccidial properties. The results showed that the NCs had a spherical shape within an average particle size of around 70 nm. The green-synthesized NCs were evaluated for their in vitro anticoccidial activity against Eimeria spp. The findings showed that the NCs exhibited a significant anticoccidial effect, with a maximum inhibition of 55.3 ± 0.32% observed at a concentration of 0.5 mg/mL. The exposure to the NCs resulted in notable alterations in the ultrastructure of the oocysts when compared to the control group. The ZnO/CuO NCs synthesized from the parsley leaf extract showed promising potential against coccidiosis and could be used in biomedical applications. Further investigation using an in vivo model is required to ascertain the efficacy of NCs as anticoccidial agents.

Opuntia Ficus-Indica Peel By-Product as a Natural Antioxidant Food Additive and Natural Anticoccidial Drug.

The present study was carried out to valorize the Opuntia ficus-indica (OFI) by-products by extracting and identifying their biochemical compounds and evaluating their antioxidant potential by in vitro activities (DPPH radical and FRAP), as well as their capacity to stabilize margarine oxidation (rancimat test). In addition, their in vitro anticoccidial effect on the destruction of Eimeria oocysts isolated from naturally infected chickens was also targeted. Microwaves and response surface methodology tools were used to extract the maximum amount of phenolic compounds (42.05 ± 0.46 GAE mg/g DW of total phenolic compounds in 90 s at 400 watts). Moreover, the effect of extraction factors was also studied. Eight phenolic compounds, including isorhamnetin, dihydrokaempferol, and kaempferol-3-O-rutinoside, were identified. The findings confirmed that OFI peel extract has strong antioxidant activities (DPPH radical, ferric reducing power). The rancimat test shows that OFI peel extract improves margarine stability by 3.2 h. Moreover, it has a notable destruction rate of Eimeria oocysts (30.06 ± 0.51%, LC50: 60.53 ± 0.38 mg/mL). The present investigation offers promise for the reuse of food waste as natural margarine additives, protection of the environment, and substitution of anticoccidial synthetic treatments.

Protective Effect of Litchi chinensis Peel Extract-Prepared Nanoparticles on Rabbits Experimentally Infected with Eimeria stiedae.

The present study used Litchi chinensis peel extract to synthesize silver nanoparticles (AgNPs). This technique is eco-friendly and can be performed in a single step; thus, it has attracted great attention for NPs biosynthesis. Herein, we biosynthesized AgNPs with L. chinensis peel extract and examined their anticoccidial activity in rabbit hepatic coccidiosis induced by E. stiedae infection. Thirty-five rabbits were allocated into seven groups: a healthy group (G1), an infected control group (G2), four groups infected before treatment with 10 mg/kg L. chinensis peel extract-biosynthesized AgNPs (G3, G5) or 50 mg/kg amprolium (G4, G6), and rabbits infected after two weeks of pretreatment with 10 mg/kg L. chinensis eel extract-biosynthesized AgNPs (G7). In this study, both pre-and post-treatment with AgNPs produced a substantial reduction in fecal oocyst output, liver enzyme levels, and histopathological hepatic lesions relative to the infected group. In conclusion, L. chinensis peel extract-prepared AgNPs should be considered harmless and efficient in the cure of hepatic coccidiosis in rabbits.

Anticoccidial effects of Tinospora rumphii extract in naturally infected goats.

Objective: This study aimed to determine the anticoccidial potential of Tinospora rumphii ethanolic stem extract, resulting in an effective reduction in oocysts per gram counts of Eimeria species in goats in comparison to the commercial coccidiostat toltrazuril. Materials and Methods: Eighteen goats that were naturally infected with coccidia were identified as experimental animals. The experimental animals were grouped and were administered a single dose of T. rumphii extract at concentrations of 300, 600, 1,000, and 2,000 mg extract/kg body weight. The fecal samples were collected before treatment, during the first treatment, and every week thereafter for 28 days posttreatment. Fecal examination was carried out using a modified McMaster technique to determine the oocysts per gram of feces, and the mean efficacies of the extracts were calculated. Results: Stem extracts of T. rumphii were able to lower the oocysts per gram count than pretreatment for all concentrations, but the efficacy, in general, was still lower than toltrazuril. A significant difference existed in the efficacy of the extracts among the treatments on day 14. Administering 1,000 mg/kg extract resulted in the highest efficacy rate (95.25%) at 14 days posttreatment and was comparable to that of toltrazuril (89.89%). Conclusion: The results show that the T. rumphii stem extract has the potential to reduce the Eimeria species oocysts per gram counts of naturally infected goats.

In vitro anticoccidial activity of nanoencapsulated bromelain against Eimeria spp. oocysts isolated from goats in Kenya.

Background and Aim: The emergence of drug-resistant strains of Eimeria spp. calls for the development of novel anticoccidial drugs. Plant extracts provide a possible natural source for such drugs. This study aimed to investigate the in vitro anticoccidial activity of encapsulated bromelain (EB) in chitosan nanocarriers on Eimeria spp. oocysts isolated from goats kept by farmers in Kenya. Materials and Methods: Bromelain was extracted from the peel of ripe pineapples using standard methods. Eimeria spp. oocysts were isolated from the feces of goats using a flotation method. The inhibition of sporulation was assayed after exposing the oocysts to solutions of EB, non-EB (NEB), and diclazuril (positive control) at concentrations between 4 mg/mL and 0.125 mg/mL for 48 h. The oocysts were examined under a microscope (40x) to determine the effects of the drugs on the sporulation process. The percentage of sporulation inhibition was calculated after 48 h and the inhibition concentration 50% (IC50) was determined by probit analysis. Results: Bromelain manifested anticoccidial activity through the inhibition of the sporulation of coccidia oocysts. EB achieved inhibition with a lower dose compared with NEB. The IC50 values of diclazuril, EB, and NEB were 0.078 mg/mL, 0.225 mg/mL, and 0.575 mg/mL, respectively. There were significant differences (p<0.01) between the IC50 of EB and NEB compared with the standard treatment drug. Conclusion: This preliminary study showed that EB has anticoccidial activity supporting further evaluation at an in vivo level to develop a novel drug for the management of coccidiosis in goats.

Effect of berberine on copper and zinc levels in chickens infected with Eimeria tenella.

Berberine, a traditional Chinese medicine, was found to exhibit anticoccidial activity. However, its mechanism is unclear. Trace metals such as copper and zinc are extremely low (less than 0.01% of the total weight of the body) but play a vital role in organisms. In the present study, we investigated the effect of berberine on copper and zinc levels in chickens infected with Eimeria tenella. Firstly, our data confirmed that infected chickens with E. tenella exhibited classic impairment on the 8th day of post infection, such as weight loss and increased feed conversion. Further study showed that E. tenella infection decreased the contents of copper and zinc in the liver and serum of chickens. Berberine was similar to amprolium and significantly improved the pathogenic conditions. Berberine could restore copper and zinc imbalance caused by E. tenella in chickens to a large extent. Studies on the development of cecum lesions demonstrated that the protective effect of berberine on the intestinal cecum was similar to that of the Cu/Zn mixture. Additionally, the mRNA expression of several metal transport related genes of the chick small intestine, including zinc transporter 1, copper transporter 1 and divalent metal ion transporter 1, was elevated by the treatment with berberine. Taken together, we speculate that the anticoccidial activity of berberine may be related to the maintenance of certain metals (Cu/Zn) homeostasis by affecting mRNA expression of their transport genes. However, the mode of action of BBR on these vital metals in the chicks infected with E. tenella still needs to be further studied.

Comparative effects of Herba Cox®, a commercial herbal extract, on rabbits (Oryctolagus cuniculus) experimentally infected with Eimeria stiedae.

The objective of this study was to evaluate the efficacy of Herba Cox®, a commercial herbal compound containing extracts from Bombax malabaricum, Aegle marmelos, Anethum foeniculum, Resina salvia, Ferula asafoetida and Papaver somniferum, for the treatment of rabbit hepatic coccidiosis. Thirty rabbits were allocated into three groups (10 × 3), the G1 group served as a negative control group, G2 group (positive control group) was infected with 5 × 104 sporulated E. stiedaeoocysts and served as infected-untreated group, and G3 group was infected with 5 × 104 sporulated E. stiedaeoocysts and treated with Herba Cox®, 1 ml/liter of drinking water, starting 7 days before infection and continuing for 4 weeks post-infection. When compared to the infected group (G2), body weight and weight gain were significantly (P ≤ 0.05) increased, the feed conversion rate was improved and no mortality was detected in infected treated group (G3) and similar to negative control group (G1). In addition, faecal oocyst output and liver enzymes were significantly decreased. Malondialdehyde, nitric oxide, and glutathione concentrations observed in G3 were similar to those in G1. In infected-untreated rabbits (G2), the haemoglobin, lymphocytes, and CD4+/ CD8+ ratio were significantly decreased, while the total leukocyte count, percentage of heterophils, and heterophil/lymphocyte ratio were increased. Significantly more severe histopathological hepatic lesions were observed in G2 when compared to G1 and G3. In conclusion, the obtained results showed that Herba Cox® should be considered a safe and novel effective compound for the treatment of E. stiedae infection in rabbits.

In Vitro Anticoccidial Activity of Olive Pulp (Olea europaea L. var. Chemlal) Extract Against Eimeria Oocysts in Broiler Chickens.

AIM: The objective of the present study was to investigate in vitro anticoccidial effect of olive pulp (Olea europaea L var. Chemlal) extract on the destruction of Eimeria spp. oocysts isolated from infected chickens naturally. MATERIALS AND METHODS: The olive pulp (OP) powder was stirred manually in aqueous ethanol in preparation for extraction using the microwave-assisted extraction system. The identification of the phenolic compounds was obtained by ultra-high-performance liquid chromatography-mass spectrometry with electrospray ionisation (HPLC-ESI-MS). The treatment of Eimeria oocyst with OP extract and standard compounds (quercetin and oleuropein) leads to their lysis as shown by the release of substances absorbing at 273 nm. RESULTS: Our results showed that the maximum number of reduced oocysts was recorded after 8 h of incubation of optimum OP extract, quercetin and oleuropein for different periods of time. Also, the number of Eimeria oocysts decreased considerably with increase concentrations after adding the optimum of OP extract in concentration ranging from 0.023 to 0.371 mg/ml. Positive correlation between the optimum OP extract concentrations and the number of Eimeria oocysts reduced was R2 = 0.959. From this in vitro experiment, it can be concluded that the OP extract possesses an anti-Eimeria spp activity. CONCLUSION: To our knowledge, this is the first time that quercetin and oleuropein were tested to evaluate their anticoccidial activity. The findings of this study showed that phenolic compound of OP extract tested separately possesses anti-Eimeria spp. effect. Further studies should be carried out to test its in vivo efficacy of the OP bioactive compounds in broiler chickens.

Anticoccidial effects of the Plantago asiatica extract on experimental Eimeria tenella infection.

Anticoccidial effects of the Plantago asiatica extract (PAE) were evaluated in chickens following oral infection with Eimeria (E.) tenella. This study was conducted on the 3-day-old chickens (n=30). Those animals were divided with 3 groups; PAE 0.1% treated/infected (n=10), PAE untreated/infected (n=10) and non-infected control (n=10). Chickens were fed a standard diet supplemented with or without PAE for 1 week prior to infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of PAE on E. tenella infection were assessed by two parameters; fecal oocysts shedding and body weights gain. The PAE-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. tenella-infected group fed standard diet. Also, PAE-based diet, improved body weight loss caused by E. tenella infection. Our data demonstrated that PAE had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of anticoccidial drug. This study is the first to demonstrate anticoccidial effect of PAE on Eimeria parasites.