Effects of Artemisia absinthium on broiler chicken coccidiosis: a systematic review and meta-analysis.

Coccidiosis is a recurring disease in broiler flocks that causes significant economic losses. This study aims to evaluate the effect of Artemisia absinthium on coccidiosis in broilers through a systematic review and meta-analysis. The article selection process included a search from the year 2000 to February 2021, with no restrictions on country or geographical region. Our objective was met by only six studies, which underwent systematic review. The meta-analysis was conducted using the metafor package in R via RStudio (version 1.1.383; RStudio, Inc.). The systematic review indicates that in vivo studies have shown the effectiveness of various plant extracts (essential oil and methanolic extract) when administered in food or drinking water on the considered parameters (oocyst shedding, bloody diarrhoea, mortality rate, weight gain, conversion index, lesion score). Furthermore, in vitro studies demonstrated a positive impact on oocyst count, LC50 (lethal concentration), sporulation rate (%), and sporulation inhibition rate (%). The meta-analysis of the four studies included in this analysis revealed that the inclusion of A. absinthium extract resulted in a significant reduction in oocyst shedding (SMD = -1.64, 95% CI: -2.72 to -0.55; P < 0.0001). However, the effectiveness of A. absinthium extract was not as significant as that of antibiotics (SMD = 0.57, 95% CI: -0.19 to 0.95; P = 0.0032). Various forms of administration and extracts of A. absinthium have demonstrated antiparasitic activity against Eimeria spp, making them suitable as natural anticoccidial agents.

Efficacy of dietary supplements of Glycyrrhiza glabra (Licorice) and maduramicin alone or in combination with Eimeria tenella infected chicks: A clinical study and molecular docking.

Background: Coccidiosis is one of the most economically significant poultry diseases worldwide, caused by the pathogenic Eimeria species, and is characterized by decreased weight gain (WG) and failure to grow due to malabsorption, low feed conversion rate, bloody diarrhea, and dehydration. Aim: This study investigated the effectiveness of licorice root extract (LRE) in controlling cecal coccidiosis to determine whether its combination with maduramicin could help alleviate the pathological, biochemical, and histopathological effects of cecal coccidiosis in Sasso broiler chicks. Methods: A total of 125 one-day-old Sasso broiler chicks were categorized into five equal groups (n = 25), each consisting of five replicates (n = 5 per replicate). G1-LE received a basal diet supplemented with LRE (3 g/kg); G2-ME received a basal diet containing maduramycin (0.5 g/kg); and G3-LME received a basal diet containing LRE and maduramicin together with the same rates. G4-E (positive control) and G5-N (negative control) received no additives in their feed. Birds in groups (G1-4) were challenged on day 14 of the experiment by orally intercropping a 1 ml suspension of Eimeria tenella sporulated oocysts. Results: Groups of birds fed on LRE and maduramicin separately or together appeared to be in good condition where no deaths or clinical abnormalities were observed, based on the analysis of clinicopathological examination. Compared with the G4-E positive control, the dropping scoring and oocyst shedding of groups G1-LE, G2-ME, and G3-LME along the 10th-day post-challenge (dpc), as well as macroscopic and microscopic lesions scoring at the 7th dpc, was considerably lower. The dual supplementation use of LRE and maduramicin in G3-LME's reduced the harmful effects of coccidian, which appeared only as a mononuclear cellular infiltration and a small number of oocysts invading the intestinal glands. Molecular docking revealed that LRE and maduramicin interacted with E. tenella DNA polymerase, E. tenella apical membrane antigen 1, and microneme protein binding sites resulting in reduced E. tenella replication and invasion. Conclusion: The inclusion of LRE and maduramicin, individually or in combination, in the diet might effectively mitigate the detrimental effects of coccidiosis.

Prophylactic and Therapeutic Efficacy of Ultrasonicated Rosmarinus officinalis Ethanolic Extract and its Chitosan-Loaded Nanoparticles Against Eimeria tenella Infected Broiler Chickens.

PURPOSE: The in vivo efficacy of ultrasonicated Rosmarinus officinalis ethanolic extract (UROEE) and its chitosan-loaded nanoparticles (UROEE-CsNPs) was investigated as a dietary prophylactic agent and as a therapeutic treatment against Eimeria tenella infected broiler chickens. METHODS: Chickens were infected with 4 × 104 E. tenella oocysts at 21 days old for primary infection and with 8 × 104 oocysts at 35 days old for secondary infection. Eleven experimental groups were conducted. Dietary addition of 100 mg/kg UROEE and 20 mg/kg for CsNPs as well as UROEE-CsNPs were included for prophylactic groups from day 1 to 42. The same doses were used for therapeutic treatment groups for 5 constitutive days. Oocyst output in feces was counted. Histopathological and immunohistochemical studies were conducted. Gene expression of pro-inflammatory cytokines as IFN-γ, IL-1ß and IL-6 as well as anti-inflammatory cytokines as IL-10 and TGF-ß4 was analyzed using semi-quantitative reverse transcriptase-PCR. RESULTS: The results showed an efficacy of UROEE, CsNPs and UROEE-CsNPs in reduction of oocyst excretion and improving the cecal tissue architecture. CD4+ and CD8+ T lymphocytes protein expression were reduced. E. tenella infection lead to upregulation of pro-inflammatory cytokines as IFN-γ, IL-1ß, IL-6 and anti-inflammatory cytokines as TGF-ß4 following primary infection, while their expression was downregulated following secondary infection. CONCLUSION: The dietary prophylactic additives and therapeutic treatments with UROEE, CsNPs and UROEE-CsNPs could decrease the inflammatory response to E. tenella as indicated by oocyst output reduction, histopathological improvements, CD4+ and CD8+ T cells protein expression reduction as well as reducing mRNA expression levels of the tested cytokines following primary and secondary infections. Consequently, these results will help to develop better-combating strategies for the control and prevention of coccidiosis on poultry farms as a dietary prophylactic agent or as a therapeutic treatment.

Growth performance, organs weight, intestinal histomorphology, and oocyst shedding in broiler chickens offered novel single strain Bacillus subtilis isolated from camel dung and challenged with Eimeria.

We evaluated a single strain Bacillus subtilis BS-9 direct-fed microbial (BSDFM) isolated from camel dung in Eimeria challenged broiler chickens. Seven-hundred d-old Ross 708 male chicks were placed in pens (25 birds/pen) and allocated to 2 treatments (n = 14). From d 0 to 13, control pens received untreated water (-BSDFM), and 2 treated pens received water and 2 mL x 108 colony forming unit/bird/d (+BSDFM); daily water intake (WI) was recorded. On d 9, birds in half (+Eimeria) of pens per treatment received of 1 mL of Eimeria maxima and Eimeria acervulina oocysts orally, and the other half (-Eimeria) sterile saline solution. Birds had ad libitum access to feed and a water line from d 14. Feed intake (FI), body weight (BW) and mortality were recorded for calculating BW gain (BWG) and feed conversion ratio (FCR). On d 14 and 35, samples of birds were necropsied for organ weight and intestinal measurements. Excreta samples were collected from d 14 to 19 for oocyst count. There was no treatment effect (P > 0.05) on growth performance or WI on d 0 to 9. There were interactions between BSDFM and Eimeria on d 19 (P = 0.014) and 29 (P = 0.036) BW with unchallenged +BSDFM birds being heavier than birds in the other treatments. The main effects (P < 0.05) on d 10 to 35 FI, BW, and BWG were such that +BSDFM increased and Eimeria decreased (P < 0.01) these parameters. There was interaction (P = 0.022) between BSDFM and Eimeria on d 10 to 35 FCR such that the FCR of challenged -BSDFM birds was poor than that of unchallenged counterparts, but none differed with +BSDFM birds. There was an interaction (P = 0.039) between BSDFM and Eimeria on d 14 bursa weight with challenged birds exhibiting heavier bursa than unchallenged +BSDFM birds. Eimeria reduced (P = 0.01) and BSDFM (P = 0.002) increased the villi height to crypt depth ratio. Results showed that BSDFM supplementation via water can support the growth performance of broiler chickens challenged with Eimeria and may be a strategy to reduce adverse effects of coccidiosis.

Bioefficacy of dietary inclusion of Nannochloropsis oculata on Eimeria spp. challenged chicks: clinical approaches, meat quality, and molecular docking.

Although anticoccidial drugs have been used to treat avian coccidiosis for nearly a century, resistance, bird harm, and food residues have caused health concerns. Thus, Nannochloropsis oculata was investigated as a possible coccidiosis treatment for broilers. A total of 150 1-day-old male Cobb broiler chicks were treated as follows: G1-Ng: fed a basal diet; G2-Ps: challenged with Eimeria spp. oocysts and fed basal diet; G3-Clo: challenged and fed basal diet with clopidol; G4-NOa: challenged and fed 0.1% N. oculata in diet, and G5-NOb: challenged and fed 0.2% N. oculata. Compared to G2-Ps, N. oculata in the diet significantly (P < 0.05) decreased dropping scores, lesion scores, and oocyst shedding. Without affecting breast meat colour metrics, N. oculata improved meat quality characters. At 28 days of age, birds received 0.2% N. oculata had significantly (P < 0.05) higher serum levels of MDA, T-SOD, HDL, and LDL cholesterol compared to G2-Ps. Serum AST, ALT, and urea levels were all decreased when N. oculata (0.2%) was used as opposed to G2-Ps. Histopathological alterations and the number of developmental and degenerative stages of Eimeria spp. in the intestinal epithelium were dramatically reduced by 0.2% N. oculata compared to G2-Ps. Molecular docking revealed a higher binding affinity of N. oculata for E. tenella aldolase, EtAMA1, and EtMIC3, which hindered glucose metabolism, host cell adhesion, and invasion of Eimeria. Finally, N. oculata (0.2%) can be used in broiler diets to mitigate the deleterious effects of coccidiosis.

Effects of Radix dichroae extract supplementation on growth performance, oocysts output and gut microbiota in growing lambs with coccidiosis.

Coccidiosis is an intestinal protozoan disease of sheep, that causes substantial economic losses in the industry due to its intestinal protozoan origins. Many anti-protozoan drugs including ionophores, triazines, and sulfonamides have been widely used to treat sheep coccidiosis. Still, anticoccidial resistance and drug residues in edible tissues have prompted an urgent search for alternatives. In this study, the anti-coccidial effectiveness of the Radix dichroae extract was compared to that of the conventional anti-coccidial drug diclazuril. Here, eighteen 45-day-old lambs naturally-infected with Eimeria spp. were randomly allocated in three groups: control group, Radix dichroae extract group and diclazuril group. The results showed that the body weight gain (BWG) during the treatment and withdrawal periods was considerably improved in the coccidiosis-infected sheep treated with Radix dichroae extract and diclazuril compared to the control group, respectively. Additionally, the Radix dichroae extract and diclazuril had fewer oocysts per gram (OPG) than the control group, showing similar anti-coccidial effects on days 14, 21, 28, 35 and 78, respectively. Furthermore, Radix dichroae extract and diclazuril treatment altered the structure and composition of gut microbiota, promoting the relative abundance of Actinobacteriota, Firmicutes, Alistipes, and Bifidobacterium, while decreasing the abundance of Bacteroidota, Marinilaceae, Helicobacteraceae, and Prevotella. Moreover, Spearman's correlation analysis further revealed a correlation between the OPG and BWG and gut microorganisms. Collectively, the results indicated that Radix dichroae extract had similar anti-coccidial effects as diclazuril, and could regulate gut microbiota balance in growing lambs.

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.

Therapeutic efficacy of Citrus aurantifolia (lime) juice in experimental Eimeria tenella-infected broiler chickens.

The purpose of the present experiment was to explore the therapeutic effect of the crude fruit juice of Citrus aurantifolia (CAJ) on Eimeria tenella disorder in chickens. One hundred twenty 3-week-old Ross 308 broilers of equal sexes were assigned to six experimental groups of 20 birds each. Groups A, B, C, D, and E were experimentally infected with 20,000 sporulated Eimeria tenella oocysts. Broilers in groups A, B, and C were infected and allocated to three treatment-graded doses of C. aurantifolia fruit juice (20, 10, and 5 mL/kg body weight, respectively) which were administered orally for 7 consecutive days of the trial. Group D was infected and treated with a reference drug, Amprolium 1.5 g/L of drinking water (positive control), group E served as infected-untreated control, and group F was uninfected and non-treated (negative control). Oocysts per gram of feces were counted using the McMaster counting device, weight gain was calculated, and blood samples from each experimental group were collected on days 0, 7, 14, 21, and 28 post-infection for hematological evaluation. Results revealed that medication of broilers with C. aurantifolia fruit juice dose-dependently increased body weight, improved cecal lesion, decreased the E. tenella oocyst production rate, and significantly (P<0.05) increased the PCV of the infected broilers. C. aurantifolia provided valuable therapeutic effects against E. tenella-induced coccidiosis in broiler chickens. The plant fruit juice should be further validated to explore the vital compounds responsible for the anticoccidial efficacy.

Persea americana extract protects intestinal tissue from Eimeria papillata-induced murine Infection.

Coccidiosis is the most prevalent disease-causing widespread economic loss among farm and domestic animals. Currently, several drugs are available for the control of this disease but resistance has been confirmed for all of them. There is an urgent need, therefore, for the identification of new sources as alternative treatments to control coccidiosis. The present work aimed to study the effect of the Persea americana extract (PAE) as an anti-coccidial, anti-oxidant, and anti-apoptotic modulator during murine intestinal Eimeria papillata infection. A total of 25 male mice were divided into five groups, as follows: Group1: Non-infected-non-treated (negative control), Group2: Non-infected-treated group with PAE (500 mg/kg b.w). Group3: Infected-non-treated (positive control), Group4: Infected-treated group with PAE (500 mg/kg b.w.), and Group5: Infected-treated group with Amprolium (120 mg/kg b.w.). Groups (3-5) were orally inoculated with 1 × 103 sporulated E. papillata oocysts. After 60 min of infection, groups (4 and 5) were treated for 5 consecutive days with the recommended doses of PAE and amprolium. The fact that PAE has an anti-coccidial efficacy against intestinal E. papillata infection in mice has been clarified by the reduction of fecal oocyst output on the 5th day post-infection by about 85.41%. Moreover, there is a significant reduction in the size of each parasite stage in the jejunal tissues of the infected-treated group with PAE. PAE counteracted the E. papillata-induced loss of glutathione peroxidase (GPx), superoxide dismutase (SOD), and total antioxidant capacity (TCA). E. papillata infection also induced an increase in the apoptotic cells expressed by caspase-3 which modulated after PAE treatment. Moreover, the mRNA expression of the goblet cell response gene, mucin (MUC2), was upregulated from 0.50 to 1.20-fold after treatment with PAE. Based on our results, PAE is a promising medicinal plant with anti-coccidial, anti-oxidant, and anti-apoptotic activities and could be used as a food additive.

Supplementing broiler diets with bacterial selenium nanoparticles enhancing performance, carcass traits, blood indices, antioxidant status, and caecal microbiota of Eimeria tenella-infected broiler chickens.

Nanomedicine is a critical therapeutic approach for treating most poultry illnesses, particularly parasitic infections. Coccidiosis is a severe protozoan infection affecting poultry; the emergence of drug-resistant Eimeria strains demands the development of new, safe therapies. Consequently, the objective of this work was to investigate the efficacy of the biosynthesized selenium nanoparticles (SeNPs) by Paenibacillus polymyxa (P. polymyxa) against Eimeria tenella (E. tenella) experimental infection in broiler chickens. The prepared SeNPs absorbed the UV at 270 nm were spherical with a size of 26 nm, and had a surface negative charge of -25 mV. One hundred and fifty, 1-day-old male broiler chicks were randomly allocated into 5 groups (30 birds/group with triplicates each) as follows: T1: negative control (noninfected and nontreated with SeNPs); T2: delivered SeNPs (500 µg/kg diet) for 35 successive days, T3: E. tenella-infected (positive control birds), T4: E. tenella-infected and treated with SeNPs (500 µg/kg diet) and T5: E. tenella-infected chicks and treated with anticoccidial agent (sulfadimidine, 16% solution 8 mL/L of drinking water) for 5 successive days. At 14 d of age, each bird in infected groups was orally treated with 3 × 103 sporulated oocyst of E. tenella. SeNPs considerably decreased the number of oocysts in broiler feces compared to positive control and anticoccidial drug, followed by a substantial reduction of parasite phase count in the cecum (15, 10, and 8 for meronts, gamonts, and developing oocysts) when compared with positive control birds. The Eimeria experimental infection lowered the activity of antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx), and reduced glutathione (GSH) while increasing the stress parameters nitric oxide (NO) and malonaldehyde (MDA). Moreover, the production of proinflammatory (TNF-α and IL-6) and apoptotic genes (BcL2 and Cas-3) were significantly elevated. Administrating SeNPs to chicks significantly decreased oxidative stress, inflammation, and apoptotic markers in the cecum tissue. Therefore, growth performance, carcass weights, antioxidant enzymes, and blood properties of infected chicks were enhanced. The findings compared the protecting role of Se-nanoparticles against cecum damages in E. tenella-infected broilers.

Comparative efficacy of allopathic and herbal drugs in sheep naturally infected with coccidiosis.

Development of anticoccidial resistance and concerns of drug residues have prompted the evaluation of alternatives to allopathic drugs. In current study, anticoccidial effect of amprolium was compared with that of Curcuma longa and Zingiber officinale. Ninety (90) sheep, naturally infected with Eimeria spp. and having a minimum oocyst per gram (OPG) count of faeces above 5000 were randomly selected and divided into six groups of 15 animals each. Animals were supplemented with amprolium @ 62.50 mg/kg body weight (bw) (GI), turmeric @ 200 and 300 mg/kg bw (GII and GIII) and ginger @ 200 and 300 mg/kg bw (GIV and GV), orally for 7 days and GVI animals were kept as untreated infected control. Faecal samples were collected on '0' day before treatment and on 8th, 14th, 21st and 28th day after starting treatment and evaluated using Faecal oocyst count reduction test (FOCRT). The efficacy of amprolium was 93.18%, 96.82%, 95.56% and 95.80% on 8th, 14th, 21st and 28th day, after starting treatment. Turmeric @200 mg/kg b.w. showed efficacy of 41.49%, 52.37%, 61.47% and 60.08% and turmeric @ 300 mg/kg bw was 44.92%, 54.32%, 64.21% and 61.95% effective on 8th, 14th, 21st and 28th day, respectively. Ginger @200 mg/kg bw showed efficacy of 38.51%, 53.48%, 55.38% and 55.53% and ginger @ 300 mg/kg bw was 39.65%, 54.81%, 57.18% and 58.22% effective on 8th,14th, 21st and 28th day, respectively. The results justify use of amprolium for clinical coccidiosis while Curcuma longa and Gingiber officinale could be used as natural prophylactic alternatives.

In vitro studies for antiparasitic activities of Punica granatum extract.

Because of the drug resistance, medicinal plants are used more frequently than coccidiostats to treat and control coccidiosis. Punica granatum is a powerful antioxidant with a variety of medicinal uses. This study used an in vitro experiment to investigate how different P. granatum from Yemen (Y) and Egypt (E) sources affected oocyst sporulation and served as an anthelminthic effector. In contrast to PGE and mebendazole, PGY (200 mg/mL) has the shortest time to paralyze and death the earthworm Eisenia fetida in this investigation. In addition, the treated worm groups' cuticle thickness and shrinkage in comparison to the control group were assessed and contrasted. Eimeria papillata is used as a model protozoan parasite in anticoccidial assays. This study shows that P. granatum affects oocysts sporulation in a dose-dependent manner, with maximal percentages of 100% (PGY) and 48.60% (PGE) at 96 h for P. granatum concentrations of 200 mg/mL. Inhibition (%) was compared to various detergents, as well as positive and negative controls. According to our research, the P. granatum extract had powerful anthelmintic and anticoccidial properties, with the potency changing according to the environmental conditions of each fruit source. RESEARCH HIGHLIGHTS: Habitat of the plant is useful for production and accumulation of some secondary metabolites in plants which be effective for the therapeutic uses. Different parameters in the environmental ecosystem affecting variation in chemical compositions and biological activity of P. granatum.

Graded levels of Eimeria challenge altered the microstructural architecture and reduced the cortical bone growth of femur of Hy-Line W-36 pullets at early stage of growth (0-6 wk of age).

An experiment was carried out to evaluate the impact of mixed Eimeria challenge on skeletal health of Hy-Line W-36 pullets. A total of 540, 16-day-old pullets were randomly allocated into 5 treatment groups, including a nonchallenged control. A mixed Eimeria species solution containing 50,000 E. maxima, 50,000 E. tenella, and 250,000 E. acervulina oocysts per mL was prepared and challenged to 1 group as a high-dose treatment. The 2-fold serial dilution was done to prepare the medium-high (25,000 E. maxima; 25,000 E. tenella; 125,000 E. acervulina), the medium-low (12,500 E. maxima; 12,500 E. tenella; 62,500 E. acervulina), and the low (6,250 E. maxima; 6,250 E. tenella; 31,250 E. acervulina) dose treatments which were challenged to 3 corresponding groups, respectively. The mineral apposition rate (MAR) was measured from 0 to 14 d post inoculation (DPI) and 14 to 28 DPI using calcein injection. The microstructural architecture of the femur was analyzed using the Skyscan X-ray microtomography (microCT) on 6, 14, and 28 DPI. The results showed that the MAR decreased linearly with an increase in the challenged dose (P < 0.05) during 0 to 14 DPI. The results of microCT revealed that cortical and total BMD, BMC, bone volume (BV), and bone volume as a fraction of tissue volume (BV/TV) of femur decreased both linearly (P < 0.05). Conversely, the total number of pores increased linearly with an increase in challenge dosages on 6 and 14 DPI. Trabecular BMD, BV, BV/TV, trabecular number, and trabecular thickness decreased linearly with an increase in the challenge dosages (P < 0.05) on 6 DPI. Furthermore, Eimeria infection significantly increased the number of osteoclasts and osteoclastic activity (P = 0.001). The result of this study suggests that the mixed Eimeria challenge negatively impacts the quality of skeletal health in a linear or quadratic manner with an increase in the concentration of Eimeria oocysts. The negative impact on long bone development might be due to malabsorption, nutrient deficiency during the infection, along with oxidative stress/inflammation disrupting the balance of osteoblastic and osteoclastic cells and their functions.

In vivo anticoccidial, antioxidant, and anti-inflammatory activities of avocado fruit, Persea americana (Lauraceae), against Eimeria papillata infection.

Apicomplexan parasites, especially Eimeria sp., are the main intestinal murine pathogens, that lead to severe injuries to farm and domestic animals. Many anticoccidial drugs are available for coccidiosis, which, leads to the development of drug-resistant parasites. Recently, natural products are considered as an alternative agent to control coccidiosis. This study was designed to evaluate the anticoccidial activity of the Persea americana fruit extract (PAFE) in male C57BL/6 mice. A total of 35 male mice were divided into seven equal groups (1, 2, 3, 4, 5, 6, and 7). At day 0, all groups except the first group which served as uninfected-untreated control were infected orally with 1 × 103E. papillata sporulated oocysts. Group 2 served as uninfected-treated control. Group 3 was considered an infected-untreated group. After 60 min of infection, groups 4, 5, and 6 were treated with oral doses of PAFE aqueous methanolic extract (100, 300, and 500 mg/kg of body weight, respectively). Group 7 was treated with amprolium (a reference drug for coccidiosis). PAFE with 500 mg/kg, was the most effective dose, inducing a significant reduction in the output of oocysts in mice feces (by about 85.41%), accompanied by a significant decrease in the number of the developmental parasite stages and a significant elevation of the goblet cells in the jejunal tissues. Upon treatment, a significant change in the oxidative status due to E. papillata infection was observed, where the levels of glutathione (GSH) increased, while, levels of malondialdehyde (MDA) and nitric oxide (NO) were decreased. In addition, the infection significantly upregulated the inflammatory cytokines of interleukin-1ß (IL-1ß), tumor necrosis factor-alpha (TNF-α), and interferon-γ (IFN-γ). This increase in mRNA expression of IL-1ß, TNF-α, and IFN-γ was about 8.3, 10.6, and 4.5-fold, respectively, which significantly downregulated upon treatment. Collectively, P. americana is a promising medicinal plant with anticoccidial, antioxidant, and anti-inflammatory activities and could be used for the treatment of coccidiosis.

Reduction of oocyte shedding and cecal inflammation by 5-aminolevulinic acid daily supplementation in laying hens infected with Eimeria tenella.

The present study aimed to evaluate the effects of 5-aminolevulinic acid (5-ALA) on Eimeria tenella infection in laying hens. Oocyst shedding and histopathology were evaluated. A reduced oocyst shedding was observed 5 and 7 days post-infection (dpi) in the 5-ALA-administered group, but the total number of oocysts during the first infection period was not different between control and 5-ALA-treated groups. After E. tenella attack infection, the period of oocyst shedding in the 5-ALA-administered group lasted less long than that in controls. During the attack infection period, the total number of fecal oocysts in the 5-ALA-treated group was significantly lower than that in the control group. However, the parasite burden score in hens receiving 5-ALA was higher than that in controls after E. tenella attack infection. The lesion scores at 5 and 30 dpi in the control group were significantly lower than those in the 5-ALA-administered group. Therefore, 5-ALA administration might be beneficial against E. tenella infection in laying hens.

Dietary organic zinc and probiotic alleviate induced Eimeria tenella infection in Japanese quails model of coccidiosis.

The current study was conducted to evaluate the impact of organic zinc (OZn) and probiotic on growth performance, oocysts number, and histological features of cecum of quails following Eimeria tenella challenge. A total of 480 Japanese quails were distributed into six treatments as follows: untreated uninfected; untreated infected; E. tenella challenge + amprolium; E. tenella challenge + OZn; E. tenella challenge + probiotic; and E. tenella challenge + OZn + probiotic. Except untreated uninfected, all groups were orally gavaged at day 8 with 5 × 104 E. tenella sporulated oocysts. Supplementation of OZn + probiotic improved (P = 0.001) growth performance compared to the untreated infected group. Lesion score of intestine and mortality was lower (P < 0.01) in quails supplemented with OZn + probiotic. Moreover, oocysts per gram (OPG) and histological dimensions of cecum in challenged birds were alleviated in OZn + probiotic. The histological findings of quails supplemented with OZn + probiotic showed normal intestinal villi with gentle sloughed epithelium. We concluded that OZn + probiotic may be safely included in the diet of Japanese quails to control coccidiosis.

In Vitro Study to Evaluate the Efficacy of Ultrasonicated Ethanolic Extract of Rosmarinus officinalis and its Chitosan-Based Nanoparticles Against Eimeria tenella Oocysts of Chickens.

In this study, chitosan nanoparticles (CsNPs) were used as nanocarrier for ultrasonicated ethanolic extract of Rosmarinus officinalis (UEERO) as a new nanoformulation against Eimeria tenella. Herein, CsNPs have been synthesized by ionic gelation method at pH 3 (CsNPs3) and pH 5 (CsNPs5), followed by characterization of morphology, size, polydispersity index (PDI), surface charge, and loading efficiency of UEERO. An in vitro sporulation inhibition assay (10, 5, 2.5, 1.25, 0.62, 0.31, 0.15, 0.07, 0.04, 0.02, and 0.01 mg/ml normal saline solution) against E. tenella was conducted. Results showed that free CsNPs and UEERO-CsNPs3/5 were cubic- and spherical-shaped with positive charge and average size of ~ 150.8 nm (314.4 nm) and 151.7 nm (321.1 nm), respectively. The total loading efficiency using UV-vis spectrophotometer, was 80.05 at pH 5 and 64.39% at pH 3. The in vitro sporulation inhibition assay revealed that UEERO, CsNPs3/5, and UEERO-CsNPs3/5 showed a potential inhibitory effect on sporulation (%), distortion in wall (%), and sporocyst abnormality (%) in a dose-dependent manner. Accordingly, the concentration (10 mg/ml) showed the best efficacy after 24 h in UEERO, free CsNPs, and UEERO-CsNPs. Moreover, UEERO-CsNPs3 and UEERO-CsNPs5 had stopped the sporulation (%) after 72 h. Taken all together, UEERO-CsNPs3 and UEERO-CsNPs5 are best effective against E. tenella in a dose-dependent manner in terms of sporulation (%), distortion in wall (%), and sporocysts abnormality.

Anti-Cryptosporidium efficacy of Citrus sinensis peel extract: Histopathological and ultrastructural experimental study.

BACKGROUND: Cryptosporidiosis is a crucial zoonotic global health concern which can be treated by alternative medicinal plants extracts. AIM OF THE STUDY: The study was carried out to assess the therapeutic efficacy of Citrus sinensis peel ethanolic extract on Cryptosporidium-infected mice. METHODS: Two doses of Citrus sinensis extract; high dose (30 mg/kg) and low dose (15 mg/kg) were investigated compared to the common commercial drug nitazoxanide (NTZ). Assessment of the extract was carried out by calculating oocysts count in fecal samples, in addition to histopathological and electron microscopic examination of intestinal mucosa.. RESULTS: There was a statistically significant reduction in the percentage of oocyst shedding more in high dose than low dose Citrus-treated mice group till negligible numbers of oocysts were found at day 22nd post infection. Histopathologically, the intestinal tissues from high dose Citrus-treated group showed improvement of the pathological changes, the villi retained their normal appearance with minimal inflammatory cells in comparison to infected control mice groups. Also, ultra-structurally, the high dose Citrus-treated mice showed few Cryptosporidium trophozoites, while moderate number of parasitic stages and mucous in the low dose Citrus-treated mice, and large numbers of parasitic stages with sever mucous in the control infected non-treated mice epithelium. CONCLUSION: Our study established for the first time that Citrus sinensis is a promising natural candidate that could be efficiently used for developing of new anti-cryptospordial drugs.

Application of CT Ultrasonography Combined with Microscopic Intraperitoneal Hyperthermic Perfusion Chemotherapy in Postoperative Treatment of Oocyst Carcinoma.

For the postoperative treatment of oocyte carcinoma, CT and CT ultrasonography combined with microscopic intraperitoneal should be combined with peritoneal heat perfusion chemotherapy. The authors selected 50 patients who received treatment for ovarian cancer from 2017 to 2018 and divided them into two groups (observation group, 30 cases, control group, 20 cases). Cisplatin, associated with peritoneal hyperthermia, led the control group, and we monitored all patients for 1 year and provided clinical trials, lifestyle, and results for both disease group. The experimental results showed that the target reduction rate of the control group was 70%, while that of the control group was only 40%, lower than that of the control group P < 0.05. The life expectancy of the control group was higher than that of the control group, P < 0.05. Nausea, vomiting, diarrhea, bone marrow compression, and constipation in the study group were slightly higher than those in the control group (35%), but there was no significant difference between the two groups (P > 0.05). Chemotherapy combined with intraperitoneal infusion of loplatin has no side effects, helps improve survival, and can be used in a variety of clinical trials.

Co-administration of chicken IL-7 or NK-lysin peptide 2 enhances the efficacy of Eimeria elongation factor-1α vaccination against Eimeria maxima infection in broiler chickens.

This study was conducted to develop a recombinant Eimeria elongation factor-1α (EF-1α)-vaccination strategy against Eimeria maxima (E. maxima) infection by co-administering with chicken IL-7 (chIL-7) or chicken NK-lysin peptide 2 (cNK-2) in commercial broiler chickens. Chickens were divided into the following 5 groups: control (CON, no Eimeria infection), nonimmunized control (NC, PBS plus Montanide ISA 78 VG), Vaccination 1 (VAC1, 100 µg of recombinant EF-1α plus Montanide ISA 78 VG), Vaccination 2 (VAC2, VAC1 plus 1 µg of chIL-7), and Vaccination 3 (VAC3, VAC2 plus 5 µg of cNK-2 peptide). The first immunization except the cNK-2 injection was performed intramuscularly on day 4, and the secondary immunization was given with the same concentration of components as the primary immunization 1 wk later. All chickens except the CON group were orally inoculated with freshly prepared E. maxima (1.0 × 104 oocysts per chicken) oocysts on Day 19. The results of the in vivo vaccination trial showed that chickens of all groups immunized with recombinant EF-1α antigen (VAC1, VAC2, and VAC3) showed higher serum antibody levels to EF-1α, and co-injection with chIL-7 further increased the serum IL-7 level in the VAC2 and VAC3 groups. Chickens in the VAC2 group showed significantly (P < 0.01) higher body weight gains at 6 and 9 d post-E. maxima challenge infection (dpi) with reduced gut lesions in the jejunum at 6 dpi. The VAC3 group showed reduced fecal oocyst shedding compared to the nonimmunized and infected chickens (NC). At 4 dpi, E. maxima infection significantly (P < 0.05) up-regulated the expression levels of proinflammatory cytokines (IL-ß and IL-17F) and type Ι cytokines (IFN-γ and IL-10) in the jejunum (NC), but the expression of these cytokines was significantly (P < 0.05) down-regulated in the VAC1, VAC2, and VAC3 groups. Furthermore, E. maxima challenge infection significantly (P < 0.05) down-regulated the expressions of jejunal tight junction (TJ) proteins (Jam2 and Occludin) at 4 dpi, but their expression was up-regulated in the VAC2 and VAC3 groups. Collectively, these results show the protective effects of the EF-1α recombinant vaccine, which can be further enhanced by co-injection with chIL-7 or cNK-2 peptide against E. maxima infection.