Expression of Toll-like receptors and host defence peptides in the cecum of chicken challenged with Eimeria tenella.

Chicken coccidiosis, caused by Eimeria protozoa, affects poultry farming. Toll-like receptors (TLRs) and host defence peptides (HDPs) help host innate immune responses to eliminate invading pathogens, but their roles in Eimeria tenella infection remain poorly understood. Herein, 14-day-old chickens were treated orally with 50,000 E. tenella oocysts and the cecum was dissected at different timepoints. mRNA expression of 10 chicken TLRs (chTLRs) and five HDPs was measured by quantitative real-time PCR. chTLR7 and chTLR15 were upregulated significantly at 3 h post-infection while other chTLRs were downregulated (p < .05). chTLR1a, chTLR1b, chTLR2b and chTLR4 peaked at 36 h post-infection, chTLR3, chTLR5 and chTLR15 peaked at 72 h post-infection and chTLR21 expression was highest among chTLRs, peaking at 48 h post-infection (p < 0.05). For HDPs, cathelicidin (CATH) 1 to 3 and B1 peaked at 48 h post-infection, liver-expressed antimicrobial peptide 2 peaked at 96 h post-infection, and CATH 2 expression was highest among HDPs. CATH2 and CATH3 were markedly upregulated at 3 h post-infection (p < .05). The results provide insight into innate immune molecules during E. tenella infection in chicken, and indicate that innate immune responses may mediate resistance to chicken coccidiosis.

The financial cost of coccidiosis in Algerian chicken production: a major challenge for the poultry sector.

Coccidiosis, caused by parasites of the genus Eimeria, is a significant economic burden to the poultry industry. In this study, we conducted a comprehensive analysis to evaluate the financial losses associated with Eimeria infection in chickens in Algeria, relying on data provided by key stakeholders in the Algerian poultry industry to assess sub-clinical as well as clinical impact. We employed the updated 2020 version of a model established to estimate the cost of coccidiosis in chickens, taking into consideration specific cultural and technical aspects of poultry farming in Algeria. The findings predict economic losses due to coccidiosis in chickens of approximately £86.7 million in Algeria for the year 2022, representing £0.30 per chicken raised. The majority of the cost was attributed to morbidity (74.9%), emphasizing the substantial economic impact of reduced productivity including decreased bodyweight gain and increased feed conversion ratio. Costs associated with control measures made up 20.5% of the total calculated cost, with 4.6% of the cost related to mortality. These figures provide a clear indication of the scope and economic impact of Eimeria infection of chickens in Algeria, illustrating the impact of practices common across North Africa. They underscore the ongoing requirement for effective preventive and control measures to reduce these financial losses while improving productivity and welfare, ensuring the economic sustainability of the Algerian poultry industry.

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.

Eimeria tenella pyrroline -5-carboxylate reductase is a secreted protein and involved in host cell invasion.

Chicken coccidiosis, which caused by Eimeria spp, is a parasitic protozoal disease. At present, control measures of this disease depend mainly on anticoccidial drugs and live vaccines. But these control strategies have drawbacks such as drug resistance and limitations in live vaccines production. Therefore, novel control approaches are urgently need to study to control this disease effectively. In this study, the function and characteristics of the pyrroline-5-carboxylate reductase of Eimeria tenella (EtPYCR) protein were preliminary analyzed. The transcription and translation level were analyzed by using qPCR and Western blot. The results showed that the mRNA transcription and translation levels of EtPYCR were higher in unsporulated oocysts (UO) and second generation merozoites (Mrz) than that in sporulated oocysts (SO) and sporozoites. Enzyme activity showed that the enzyme activity of EtPYCR was also higher in the UO and Mrz than that in the SO and sporozoites. Immunofluorescence localization showed EtPYCR was mainly located on the top of sporozoites and the whole cytoplasm and surface of Mrz. The secretion assay indicated that EtPYCR was secretion protein, but not from micronemes. Invasion inhibition assay showed that rabbit anti-rEtPYCR polyclonal antibodies can effectively inhibit sporozoite invasion of DF-1 cells. These results showed that EtPYCR possess several important roles that separate and distinct from its conversion 1-pyrroline-5-carboxylate (P5C) into proline and maybe involved in the host cell invasion and development of parasites in host cells.

Systemic Caryospora-like coccidiosis in a clutch of hatchling red-eared slider turtles (Trachemys scripta elegans).

Caryospora-like organisms (CLOs) form a clade of at least 11 genotypes of related coccidia that can cause epizootic mortality in marine turtles. The biology, transmission, host species range, and host cell tropism of these organisms are still largely unknown. The goal of this study was to characterize the host cell tropism, pathologic and ultrastructural features, and phylogeny associated with the first report of a mortality event due to CLO in the freshwater red-eared slider turtle (Trachemys scripta elegans). Sudden mortalities within a clutch of captive-raised red-eared slider hatchlings (n = 8) were recorded, and deceased animals had severe segmental to diffuse, transmural, fibrinonecrotic enterocolitis and multifocal to coalescing hepatic necrosis, among other lesions associated with numerous intracytoplasmic developing stages of intralesional coccidia. Among the different developmental stages, merozoites were ultrastructurally characterized by an apical complex. A pan-apicomplexan polymerase chain reaction (PCR) yielded a 347 bp-amplicon matching the Schellackia/Caryospora-like clade with 99.1% identity to the US3 strain from green sea turtles (Chelonia mydas) and 99.1% identity to Schellackia sp. Isolate OC116. Surviving hatchlings were treated with toltrazuril sulfone (ponazuril) but were subsequently euthanized due to the risk of spreading the parasite to other chelonids in the collection. The ponazuril-treated hatchlings (n = 4) had mild proliferative anterior enteritis, with few intraepithelial coccidia in one hatchling confirmed as CLO by PCR. This is the first report of Caryospora-like coccidiosis in non-cheloniid turtles, highlighting the relevance of this disease as an emerging highly pathogenic intestinal and extra-intestinal form of coccidiosis of turtles with potential cross-species infectivity.

On the efficacy of preventive toltrazuril treatments and the diagnosis of Cystoisospora suis infections in intensively raised piglets in farms from southeast Spain.

This study investigated the presence and abundance of Cystoisospora suis oocysts in faecal samples from 131 one- to three-week-old pig litters belonging to eight intensively raised, indoor herds in Spain. Seven herds used preventive anticoccidial toltrazuril treatments administered orally or by intramuscular injection, and one did not use preventive anticoccidial treatments. The diagnosis was performed using two oocyst flotation-concentration methods, Bailenger's method in every herd and the more recent Joachim's method in four herds. Oocysts were detected in every farm, the proportion of oocyst-positive samples was higher with Bailenger's technique, and the estimated overall prevalence (95% confidence interval) was 40 (32-49)%, including 47 (29-65)% in non-medicated litters, 52 (38-67)% in orally medicated litters and 28 (16-40)% in intramuscularly medicated litters (p < 0.05). However, mixed logistic regression models indicated that the risk of infection was not significantly associated with preventive anticoccidial treatments (p > 0.05), and increased with age, was higher in herds with partially compared to fully slatted dung floors in farrowing pens and in litters with pigs with diarrhoea (p < 0.05). The median (range) oocysts per gram of faeces (OpG) in infected litters by Bailenger's method was 623 (35-49048) and mixed negative binomial models revealed no significant association between infection intensity in positive litters and pen's floor type and piglets age, faecal consistency and treatment status (p > 0.05). The apparent low efficacy of Toltrazuril suggests treatment administration failures, reduced residual efficacy or low susceptibility of C. suis strains in study farms and needs further investigation.

Identification of sheep eimerian parasites, Eimeria crandallis and Eimeria faurei, employing microscopic and molecular tools.

Coccidiosis, caused by apicomplexan Eimeria species, is a protozoan disease that affects various species of wild and domestic animals. However, data available on Eimeria diversity in ruminants in Saudi Arabia is meagre. Therefore, this study was designed to investigate some eimerian parasites infecting sheep (Sawakni and Harrie breeds) using microscopy and molecular methods for the first time in Saudi Arabia. Twenty-four fecal samples were collected from sheep farms. Based on the floatation technique, eimerian oocysts were observed in 8 of the 24 (33.33%) fecal samples. The coccidian-positive samples were subjected to fecal culture in a shallow layer of 2.5% potassium dichromate (K2 Cr2 O7 ). Detected eimerian oocysts were described micromorphometrically as the basis for traditional oocyst identification. Morphologically, the sporulated oocysts were similar to those of sheep eimerian parasies; Eimeria faurei and Eimeria crandallis. PCR products from the two eimerian species detected from Sawakni and Harrie breeds were sequenced and were found to be distinct from each other with mutations at five positions. One of them clustered with E. crandallis with 99.8%-100% identity with sequences available in GenBank. E. crandallis was obtained from two Sawakni sheep and two Harrie sheep. The other sequences grouped with E. faurei with 99.8% identity with the only sequences available in GenBank. E. crandallis was detected from both Sawakni and Harrie breeds whereas E. faurei was detected only from Sawakni sheep. The findings of this study have implications for the importance of morphometric identification with advanced molecular tools to confirm the identities of sheep Eimeria species and to address the taxonomic study of this eimeriid parasite at the species level.

Identification of Eimeria acervulina (Apicomplexa: Eimeriidae) infecting the broiler chicken Gallus gallus domesticus through morphology and molecular analyses.

Coccidiosis is an intestinal protozoan disease that affects the poultry industry worldwide. The severity of this disease varies depending on the identity of the infectious agents. Therefore, this study was carried out to identify the Eimeria species that affect broiler chickens, Gallus gallus domesticus, through morphological and molecular phylogenetic analyses. Twenty-five faecal samples were collected from the broiler chickens in a commercial poultry farm in Riyadh (Saudi Arabia). Using the floatation technique, faeces were examined microscopically for the Eimeria occurrence. Identification of Eimeria species was performed based on morphological criteria and molecular tools (DNA amplification for the partial small subunit ribosomal RNA (18S rRNA), internal transcribed spacer (ITS)-1, and mitochondrial cytochrome c oxidase I (COI) genes. In this study, 32% (8 out of 25) of collected samples were found to be positive for coccidiosis. After sporulation in potassium dichromate (K2Cr2O7), the sporulated oocysts were observed as ovoid and measured 18.37-23.19 µm (19.87) long and 15.07-18.67 µm (16.46) wide, with the anterior location of a polar granule and absence of micropyle. These Eimeria oocysts were assumed to size and shape characteristics of Eimeria acervulina. Molecular analysis was conducted on the sequences of the polymerase chain reaction products from the three genes studied (18S rRNA, ITS-1, and COI). At the three genes, results showed that the resultant sequences clustered with E. acervulina from different regions confirming morphological description. This study highlighted the importance of molecular techniques to detect avian Eimeria species more than the traditional morphology-based tool to optimise the appropriate anticoccidial strategies for long-term control in the studied area.

Protective efficacy of multiepitope vaccines constructed from common antigens of Eimeria species in chickens.

Clinical avian coccidiosis is typically caused by coinfection with several Eimeria species. Recombinant protein and DNA vaccines have shown promise in controlling coccidiosis. On this basis, DNA vaccines that encode multiple epitopes from different Eimeria species may provide broad protection against coinfections. In this study, we designed a fusion gene fragment, 14EGT, that contained concentrated T-cell epitopes from four common antigens of Eimeria species (14-3-3, elongation factor 2, glyceraldehyde-3-phosphate dehydrogenase, and transhydrogenase). The multiepitope DNA vaccine pVAX1-14EGT and recombinant protein vaccine pET-32a-14EGT (r14EGT) were then created based on the 14EGT fragment. Subsequently, cellular and humoral immune responses were measured in vaccinated chickens. Vaccination-challenge trials were also conducted, where the birds were vaccinated with the 14EGT preparations and later exposed to single or multiple Eimeria species to evaluate the protective efficacy of the vaccines. According to the results, vaccination with 14EGT preparations effectively increased the proportions of CD4+ and CD8+ T cells and the levels of Th1 and Th2 hallmark cytokines. The levels of serum IgG antibodies were also significantly increased. Animal vaccination trials revealed alleviated enteric lesions, weight loss, and oocyst output compared to those of the control groups. The preparations were found to be moderately effective against single Eimeria species, with the anticoccidial index (ACI) ranging from 160 to 180. However, after challenge with multiple Eimeria species, the protection provided by the 14EGT preparations was not satisfactory, with ACI values of 142.18 and 146.41. Collectively, the results suggest that a multiepitope vaccine that encodes the T-cell epitopes of common antigens derived from Eimeria parasites could be a potential and effective strategy to control avian coccidiosis.

The role of genetics in determining resistance to coccidiosis in goats a review of current research and future directions.

Coccidiosis is a significant parasitic disease in goats, with significant impacts on animal health, productivity, and economic losses for producers. Although various management practices can help control and prevent coccidiosis, a growing body of research suggests that genetics play an important role in determining resistance to the disease. This review explores the current understanding of the genetics of coccidiosis resistance in goats, including the potential genetic factors and mechanisms involved, and the implications for breeding and selection programs. The review will also discuss current research and future directions in this field, including the use of genomic tools and technologies to better understand the genetics of resistance and to improve breeding programs for coccidiosis resistance in goats. This review will be of interest to veterinary practitioners, goat producers, animal breeders, and researchers working in the field of veterinary parasitology and animal genetics.

From celiac disease to coccidia infection and vice-versa: The polyQ peptide CXCR3-interaction axis.

Zonulin is a physiological modulator of intercellular tight junctions, which upregulation is involved in several diseases like celiac disease (CeD). The polyQ gliadin fragment binds to the CXCR3 chemokine receptor that activates zonulin upregulation, leading to increased intestinal permeability in humans. Here, we report a general hypothesis based on the structural connection between the polyQ sequence of the immunogenic CeD protein, gliadin, and enteric coccidian parasites proteins. Firstly, a novel interaction pathway between the parasites and the host is described based on the structural similarities between polyQ gliadin fragments and the parasite proteins. Secondly, a potential connection between coccidial infections as a novel environmental trigger of CeD is hypothesized. Therefore, this report represents a promising breakthrough for coccidian research and points out the potential role of coccidian parasites as a novel trigger of CeD that might define a preventive strategy for gluten-related disorders in general. Also see the video abstract here: https://youtu.be/oMaQasStcFI.

Chicken ß-defensin-1 peptide as a candidate anticoccidial agent in broiler chickens.

The current study aimed to investigate the potentiality of using avian ß-defensin-1 peptide as a candidate agent against coccidiosis infection in broiler chicken.We employed an in-silico analysis to study the primary structure of ß-defensin-1 peptide as well as its 3-D and molecular dynamic structures. This will also enable obtaining adequate information about the mode of action of these peptides and the intra-cellular transduction pathways. The results revealed no significant difference among groups of broiler chicken in terms of body weight before the Eimeria challenge.The results of our study indicated a significant reduction in oocyst count in birds administered ß-defensin-1 peptide treatment, vis-a-vis healthy birds. The treated group showed a 2-3 times reduction in oocyst count, compared to the positive control group. The Eimeria oocysts count evaluated for birds administered with ß-defensin-1 after the Eimeria challenge showed a significant difference. The study indicated significant reduction and down-regulation in the level of expression of ß-defensin 1 and 4 in the control and treatment groups.This electrostatic profile and hydrophobicity regulate the functioning of this peptide. The results may help in the development of novel approaches that could be used as alternatives or adjunct to the existing means of coccidiosis control in broilers.

Production performance and economic analysis of broiler chickens after vaccination with a live attenuated vaccine against avian coccidiosis.

The objective of this study was to evaluate the effectiveness of live attenuated commercial vaccine LIVACOX® T against avian coccidiosis upon parasite exposure through its correlation with productive and economic performance, clinical observation, and oocyst excretion of broiler chickens. For this purpose, 420 1-day-old Cobb chicks were divided into five groups of 84 birds: (G1) unvaccinated and unchallenged control; (G2) vaccinated on day 0; (G3) challenged on day 1; (G4) vaccinated on day 0 and challenged on day 14; and (G5) challenged on day 14. For 28 days, the clinical signs of infection, weight and feed conversion of the birds, and excretion of oocysts in the feces were evaluated. Macroscopic analysis of intestinal lesions in birds was also performed. After vaccination in G2, G3, and G4 as well as after challenge in G3, G4, and G5, there was an increase in oocyst excretion. In the analysis of weight gain, the difference in final weight between groups G3 and G4 is - 105.74 g per bird. Therefore, if we multiply this value by the average number of birds slaughtered per day in a medium/large slaughterhouse (250,000), we have 26,435 kg of chicken meat per day of slaughter, representing 581,570 kg of monthly losses (22 days of slaughter/month), or approximately R$3,489,420.00 (US$872,355.00), considering the commercial value at R$6.00/kg (US$ 1.5/kg). Thus, the productive and economic impact of coccidiosis in broiler chickens is evident, and the importance of vaccination to prevent the occurrence of the disease and reduce subsequent loss is highlighted.

Dietary Supplementation with Eugenol Nanoemulsion Alleviates the Negative Effects of Experimental Coccidiosis on Broiler Chicken's Health and Growth Performance.

The present study investigated the protective efficacy of dietary supplementation with clove essential oil (CEO), its main constituent eugenol (EUG), and their nanoformulated emulsions (Nano-CEO and Nano-EUG) against experimental coccidiosis in broiler chickens. To this aim, various parameters (oocyst number per gram of excreta (OPG), daily weight gain (DWG), daily feed intake (DFI), feed conversion ratio (FCR), serum concentrations of total proteins (TP), albumin (ALB), globulins (GLB), triglycerides (TG), cholesterol (CHO) and glucose (GLU), serum activity of superoxide dismutase (SOD), glutathione s-transferase (GST), and glutathione peroxidase (GPx)] were compared among groups receiving CEO supplemented feed (CEO), Nano-CEO supplemented feed (Nano-CEO), EUG supplemented feed (EUG), Nano-EUG supplemented feed (Nano-EUG), diclazuril supplemented feed (standard treatment, ST), or basal diet [diseased control (d-CON) and healthy control (h-CON)), from days 1-42. Chickens of all groups, except h-CON, were challenged with mixed Eimeria spp. at 14 days of age. Coccidiosis development in d-CON was associated with impaired productivity (lower DWG and higher DFI and FCR relative to h-CON; p < 0.05) and altered serum biochemistry (decreased TP, ALB, and GLB concentrations and SOD, GST, and GPx activities relative to h-CON; p < 0.05). ST effectively controlled coccidiosis infection by significantly decreasing OPG values compared with d-CON (p < 0.05) and maintaining zootechnical and serum biochemical parameters at levels close to (DWG, FCR; p < 0.05) or not different from (DFI, TP, ALB, GLB, SOD, GST, and GPx) those of h-CON. Among the phytogenic supplemented (PS) groups, all showed decreased OPG values compared with d-CON (p < 0.05), with the lowest value being measured in Nano-EUG. All PS groups showed better values of DFI and FCR than d-CON (p < 0.05), but only in Nano-EUG were these parameters, along with DWG, not different from those of ST. Furthermore, Nano-EUG was the only PS group having all serum biochemical values not different (or even slightly improved) relative to ST and h-CON. In conclusion, the tested PS diets, especially Nano-EUG, can limit the deleterious effects of coccidiosis in broiler chickens, due to anticoccidial activity and possibly their reported antioxidant and anti-inflammatory properties, thereby representing a potential green alternative to synthetic anticoccidials.

Synthesis of Silver Nanoparticles by Leaf Extract of Cucumis melo L. and Their In Vitro Antidiabetic and Anticoccidial Activities.

In this study, silver nanoparticles were synthesized using Cucumis melo L. leaf extract via a green synthesis approach and their potential against diabetes and coccidiosis was tested under in vitro conditions. The phytochemical components in the leaf extract reacted with silver nitrate in solution and yielded C. melo-silver nanoparticles (Cm-AgNPs). The synthesis of AgNPs was confirmed via UV-visible spectroscopy by obtaining a peak at 440 nm. The nanoparticles were characterized by their morphology, crystallinity, and the presence of functional groups. In vitro α-amylase and α-glucosidase inhibition assays were carried out at different concentrations in the range of 20 to 100 µg/mL of Cm-AgNPs. The Cm-AgNPs exhibited enzyme inhibitory activity in a concentration-dependent manner. As the concentration of Cm-AgNPs increased the inhibitory activities were also increased linearly and the highest inhibition was observed at 100 µg/mL. The effectiveness of Cm-AgNPs against Eimeria tenalla was assessed by an in vitro 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay using Madin-Darby bovine kidney (MDBK) cell lines. The results revealed that the viability of the oocysts and further sporulation were decreased with the increased concentration of Cm-AgNPs. The AgNPs synthesized from the C. melo leaf extract have shown promising potential against diabetes and coccidiosis, and they could be used in biomedical applications.

Dietary tea tree (Melaleuca alternifolia) essential oil as alternative to antibiotics alleviates experimentally induced Eimeria tenella challenge in Japanese quails.

Coccidiosis is one of the most common infectious diseases seen in Japanese quails. The current study was conducted to evaluate the impact of tea tree essential oil (TTEO) on growth performance and intestinal health of quails in response to Eimeria tenella challenge. A total of 250 Japanese quails were divided into five treatments: untreated uninfected (negative control); untreated infected (positive control); infected + Amprolium; infected and 1% TTEO; infected and 2% TTEO. Except negative control, all groups were orally dosed with 5 × 104 sporulated oocysts of E. tenella. The results revealed that supplementation of 1% TTEO and treatment of amprolium improved feed intake, weight gain and feed conversion ratio in infected quails compared to the positive control. Similarly, lesion score and mortality was significantly (p < 0.01) reduced in quails supplemented with 2% TTEO and amprolium treated birds. Moreover, oocysts counts and histological features of caecum in infected birds were reversed in 1% TTEO and amprolium treatment. The histological findings of amprolium and 1% TTEO supplemented quails showed intact intestinal villi with mild sloughed epithelium. In conclusion, 1% TTEO can be safely used to control coccidiosis in Japanese quails as natural effective compound.

High prevalence of pathogenic Eimeria spp. and the main risk factors associated with infection in goats from a semiarid region of Northeastern Brazil.

This study aimed to analyze the epidemiology of infections caused by Eimeria spp. in goats bred in an extensive production system in Northeast Brazil. To this end, an epidemiological survey was performed, wherein 350 fecal samples were collected from goats ranging from 3 months to 8 years old across 30 farms. The number of oocysts per gram of feces (OPG) was counted using the Mini-FLOTAC method. Species were identified based on their morphological and morphometric characteristics. Frequencies between animal categories were assessed using the chi-square test and known risk factors after multivariate analysis. Eimeria oocysts were found in 99.4% (346/348) of samples and 100% (30/30) of the farms. The OPG was influenced by the age of the animals; therefore, young goats (up to 6 months old) eliminated more oocysts (2302.6 + 5454.8) than adult goats (881.7 + 1551.5). Three pathogenic species were identified, with two demonstrating a high prevalence: Eimeria arloingi (88.83%) and Eimeria christenseni (44.7%), in addition to Eimeria caprina (3.72%). In the study, the habit of ground feeding and the use of cisterns as water sources were identified as risk factors, with odds ratios of 4.04 and 9.82, respectively. However, cleaning facilities at shorter intervals was a protective factor (OR = 0.19). We concluded that there was a high prevalence of Eimeria infection in goats bred in extensive production systems, even those in semiarid regions. This high prevalence can be attributed to the congregation of animals in shaded areas and the substantial accumulation of fecal matter at night.

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.

Molecular characterization and immune protection by cystathionine ß-synthase from Eimeria tenella.

Eimeria tenella is an obligate intracellular apicomplexan parasite that causes avian coccidiosis and leads to severe economic losses in the global poultry industry. Cystathionine ß-synthase (CBS) and cystathionine γ-lyase (CGL) act together to generate H2S in the reverse transsulfuration pathway. In this study, E. tenella Cystathionine ß-synthase (EtCBS) was cloned using rapid amplification of cDNA 5'-ends (5'RACE) and characterized, and its immunoprotective effects were evaluated. The recombinant EtCBS protein (rEtCBS) was expressed and successfully recognized by anti-sporozoites (Spz) protein rabbit serum. EtCBS mRNA levels were highest in Spz by qPCR, and the protein expression levels were higher in unsporulated oocysts (UO) than in other stages by Western blot. Indirect immunofluorescence showed that EtCBS protein was found on the surface of Spz and second-generation merozoites (Mrz). The invasion inhibition assays showed that rabbit anti-rEtCBS polyclonal antibodies effectively inhibited parasite invasion host cells. Chickens immunized with rEtCBS protein showed prominently increased weight gains and decreased oocyst output compared to nonimmunized and infected control group. The results suggest that EtCBS could be a potential vaccine candidate against E. tenella.

Differential expression of microRNAs in the caecal content and faeces of broiler chickens experimentally infected with Eimeria.

Coccidiosis caused by Eimeria spp. incurs significant morbidity and mortality in chickens, and is thus of great economic importance. Post-mortem intestinal lesion scoring remains one of the most common means of diagnosis; therefore alternative, non-invasive methods of diagnosis and monitoring would be highly desirable. Micro-RNAs (miRNAs) have been shown to be stable in faeces of human and animal species with expression altered in gastrointestinal disease. We hypothesized that miRNA is stable in caecal content of chickens, and that differential miRNA expression patterns would be seen in Eimeria-infected versus uninfected individuals. Initially, RNA was extracted from Eimeria tenella-infected (n = 3; 7 days post infection) and uninfected (n = 3) chicken caecal content to demonstrate miRNA stability. Subsequently, next-generation miRNA sequencing was performed on caecal content from E. tenella-infected chickens with high (lesion score (LS) 3-4; n = 3) or low (LS1; n = 3) levels of pathology, and uninfected controls (n = 3). Comparative analysis identified 19 miRNAs that exhibited significantly altered expression in the caecal content of E. tenella, infected chickens versus uninfected chickens (t-test, False Discovery Rate (FDR) < 0.05). Eight of these miRNAs showed significant up-regulation in infection (fold change of 9.8-105, FDR <0.05). Quantitative PCR was performed using separate biological replicates to confirm differential regulation in eight of these miRNA candidates in caecal and faecal content. This work has identified a panel of miRNA candidates which may be appropriate for use as non-invasive faecal markers of active caecal coccidiosis without the need for culling. RESEARCH HIGHLIGHTSE. tenella induced differential miRNA expression in caecal content and faeces.