Development of a PCR assay for detection and identification of Eimeria spp. in cattle.

Eimeria spp. are important coccidian parasites causing diarrhea and significant mortality in cattle worldwide. To date, at least 13 Eimeria species with varying pathogenicity have been identified in cattle. Efficient detection and identification of Eimeria spp. is therefore essential for the prevention and control of bovine coccidiosis. However, the commonly used microscopic examination for Eimeria spp. is time-consuming and requires considerable expertise. In this study, we aligned the nucleotide sequences of the small subunit (SSU) rRNA gene of common Eimeria species and developed a nested PCR assay targeting the polymorphic SSU rRNA region of Eimeria spp. from cattle. Initially, the SSU rRNA gene PCR assay was compared with microscopic examination for sensitivity and detection range of Eimeria species using fecal samples from dairy cattle. Of the 193 fecal samples, 131 (67.9 %) and 78 (40.4 %) were positive for Eimeria by PCR and microscopy, respectively. Sequence analysis of the PCR products identified six Eimeria species, including E. cylindrica (n = 76), E. bovis (n = 54), E. auburnensis (n = 30), E. zuernii (n = 25), E. wyomingensis (n = 10), E. canadensis (n = 1), and co-infections of 2-4 species (n = 55). In contrast, only the first four species and co-infections of 2-3 species were identified by microscopy. The PCR assay was able to detect as few as 50 Eimeria oocysts per gram of feces. Thus, the developed SSU rRNA gene PCR assay has a high sensitivity and allowed easy identification of at least six common Eimeria species and their co-infections in cattle. It should be useful in molecular epidemiological studies of bovine coccidiosis.

Mycotoxins and coccidiosis in poultry - co-occurrence, interaction, and effects.

Avian coccidiosis, a common disease caused by Eimeria species, results in significant losses in global poultry production. Mycotoxins are low-molecular-weight natural products (i.e., small molecules) produced as secondary metabolites by filamentous fungi and they have the potential to economically and significantly affect global poultry production. Little is known about the relationship between mycotoxins and avian coccidiosis, although they often co-occur in the field. This comprehensive review examines the intricate relationship between mycotoxins and avian coccidiosis, in particular how mycotoxins, including aflatoxins, ochratoxins, trichothecenes as well as Fusarium mycotoxins, compromise the health of the poultry flock and open the door to Eimeria parasites in the gut. In addition, this review sheds light on the immunosuppressive effects of mycotoxins, their disruption of cellular signaling pathways, and the consequent exacerbation of coccidiosis infections. The mechanisms of mycotoxin toxicity are also reviewed, emphasizing direct damage to intestinal epithelial cells, impaired nutrient absorption, inflammation, oxidative stress, and changes in the gut microbiota. Finally, the consequences for the prevention and treatment of coccidiosis when mycotoxins are present in the feed are discussed. This review emphasizes the need for effective management strategies to mitigate the combined risks of mycotoxins and coccidiosis and highlights the complexity of diagnosing and controlling these interrelated problems in poultry. The review advocates a holistic approach that includes strict feed management, disease prevention measures and regular monitoring to maintain the health and productivity of poultry against these significant challenges.

Trifolium pratense as a novel phytogenic supplement, is an anticoccidial agent in chickens.

Coccidiosis, caused by a protozoan parasite of the genus Eimeria, is one of the most severe contagious parasite diseases affecting the poultry industry worldwide. Using phytogenics to prevent chicken coccidiosis is a strategy aimed at combating the increasing issue of drug-resistant strains of Eimeria spp. This study demonstrates the anticoccidial activities of a medicinal herb, Trifolium pratense (TP) powder, and its ethanolic extract (designated TPE) against Eimeria spp. TPE exhibited significant suppressive activity against E. maxima oocyst sporulation and E. tenella sporozoite invasion and reproduction in Madin-Darby bovine kidney cells. Furthermore, administration of basal chicken diets containing TP powder or TPE to Eimeria-infected chickens significantly reduced the output of oocysts and severity of intestinal lesions. Dietary supplementation with TP significantly improved relative weight gain in E. tenella- and E. acervulina-infected chickens, while there was no significant improvement in E. maxima-infected chickens. The anticoccidial activities of TP and TPE on E. acervulina, E. tenella and E. maxima were further supported by anticoccidial index scores, which showed greater efficacy than those of amprolium, a commercial coccidiostat used in poultry. TP supplementation positively impacted the primary metabolism of chickens challenged with E. tenella or E. acervulina. The chemical fingerprints of TPE were established using liquid column chromatography; TPE contained 4 major compounds: ononin, sissotrin, formononetin, and biochanin A. In addition, various spectrometric methods were used to ensure the batch-to-batch consistency of TP/TPE. In conclusion, T. pratense is demonstrated to be a novel phytogenic supplement that can be used to control Eimeria-induced coccidiosis in chickens.

Epidemiological investigation of coccidiosis and associated risk factors in broiler chickens immunized with live anticoccidial vaccines in China.

Coccidiosis is a costly intestinal disease of chickens caused by Eimeria species. This infection is associated with high mortality, reduced feed efficiency, and slowed body weight gain. The diagnosis and control of coccidiosis becomes challenging due to the fact that chickens can be infected by seven different Eimeria species and often occur mixed-species co-infections. Grasping the epidemiology of Eimeria species is crucial to estimate the efficiency of poultry management. This study aimed to explore the distribution of Eimeria species in broiler chickens in China after administering live anticoccidial vaccines. A total of 634 samples were obtained, and the survey results showed that the prevalence of Eimeria was 86.12% (546/634), and the most common species were E. acervulina (65.62%), E. necatrix (50.95%), E. mitis (50.79%), E. tenella (48.42%), and E. praecox (41.80%). Most samples indicated mixed-species infections (an average of 3.29 species per positive sample). Notably, 63.98% of samples contain 3 to 5 Eimeria species within a single fecal sample. The most prevalent combinations were E. acervulina-E. tenella (38.96%) and E. acervulina-E. necatrix (37.22%). Statistical analysis showed that flocks vaccinated with trivalent vaccines were significantly positive for E. necatrix in grower chickens (OR = 3.30, p < 0.05) compared with starter chickens, and tetravalent vaccinated flocks showed that starter chickens demonstrated a higher susceptibility to E. tenella-E. brunetti (OR = 2.03, p < 0.05) and E. acervulina-E. maxima (OR = 2.05, p < 0.05) compared with adult chickens. Geographically, in the case of tetravalent vaccine-immunized flocks, a substantial positive association was observed between E. necatrix infection rates and flocks from eastern (OR = 3.88, p < 0.001), central (OR = 2.65, p = 0.001), and southern China (OR = 3.17, p < 0.001) compared with southwestern China. This study also found a positive association between E. necatrix (OR = 1.64, p < 0.05), E. acervulina (OR = 1.59, p < 0.05), and E. praecox (OR = 1.81, p < 0.05) infection and coccidiosis occurrence compared with non-infected flocks in tetravalent vaccinated flocks. This molecular epidemiological investigation showed a high prevalence of Eimeria species in the field. The emergent species, E. brunetti and E. praecox, might be incorporated into the widely-used live vaccines in the future. These insights could be useful in refining coccidiosis control strategies in the poultry industry.

Age-dependent development trends (models) of intestinal significant microbiota species and Eimeria oocysts in coccidia-challenged broiler chickens as affected by dietary encapsulated organic acids and anticoccidial drugs.

ABSTRACTThe study was conducted to investigate the effect of dietary encapsulated organic acids (EOAs) and anticoccidials on the age-dependent development trend of intestinal Lactobacillus, E. coli, coliforms, and Eimeria in Eimeria spp.-infected broiler chickens from reused litter. In total, 525 mixed-sex 1-day-old broiler chickens were used in an uninfected/un-supplemented control plus a 2 (no EOA or 0.1% EOA) × 3 (no anticoccidial, 0.05% maduramicin, and 0.02% diclazuril) factorial arrangement of treatments as a completely randomized design with five replicates of 15 chickens. Results indicated that the cubic model is the best model for explaining the development trends of the intestinal microbial population in uninfected and infected chickens (affected by the EOAs and anticoccidials). Based on the cubic models, the microbial populations had development trends with a decreasing slope from 1-day-old until the early or middle finisher period. EOAs and anticoccidials, especially their simultaneous usage, improved (P < 0.05) the linear and cubic models' slope (affected negatively by Eimeria infection). A polynomial model (order = 6) was determined as the best model for explaining the EOAs and anticoccidial effects on the trend of intestinal Eimeria oocysts in infected chickens. The infection peak (which happened at 25 days) was reduced by EOAs and anticoccidials, especially their simultaneous usage. In conclusion, cubic and polynomial (order = 6) regressions are the best models fitted for explaining the microbiota and Eimeria oocysts trends, respectively. EOAs and anticoccidials, especially their simultaneous usage, had beneficial effects on the microbiota and Eimeria development trends and gastrointestinal health in coccidia-infected broiler chickens. RESEARCH HIGHLIGHTSCubic regression is the best model for explaining intestinal microbiota development.Polynomial regression is the best model for intestinal Eimeria oocysts development.Age-development trends are affected by dietary encapsulated organic acids and anticoccidials.

Differential diagnosis of Eimeria species in farmed Japanese quails (Coturnix japonica).

Similarly to poultry industry, coccidiosis may cause significant economic losses also in the commercial quail industry, an emerging sector undergoing uneven development around the world. Although scant and mostly dated, the available literature reports detailed morphological and morphometric features of both oocysts and sporocysts of the Eimeria species hitherto recognized in Japanese quails, i.e. E. tsunodai, E. uzura, E. bateri, and E. fluminensis. Mixed infections are very common in the field and require an accurate differential diagnosis of diverse species of coccidia, identifying the highly pathogenic ones, in particular E. tsunodai (localized in the caeca), and E. uzura (localized in both caeca and small intestine). This goal is hampered by time-consuming laboratory procedures involving highly qualified staff and facilities, and poorly compatible with routine management practices in farmed quails. A supplemental difficulty is represented by the lack of nucleotide sequences available in GenBank. To overcome these issues, copromicroscopic and molecular analyses (amplifying the 18S rRNA region, and the internal transcribed spacers regions ITS1-5.8rRNA-ITS2) were performed on oocysts populations separately isolated from pools of 12 caecal and 12 cloacal contents collected from 240 naturally infected laying Japanese quails. Data on morphological and morphometric features of 1,000 sporulated oocysts were statistically compared, demonstrating the presence of different Eimeria species colonizing the 2 intestinal tracts. This result was also confirmed by PCR and phylogenetic analysis of the 18S rRNA gene. Overall results allowed to hypothesize the presence of E. uzura in our Japanese quails. Although a certain identification at species level was not obtained, the present study demonstrates that reasonable turnaround times of monitoring procedures performed on Japanese quail farms, shedding light on the in vivo and post-mortem differential diagnosis of coccidiosis can be achieved, and provide obvious benefits in disease understanding and control.

Gastrointestinal Parasite Infections and Environmental Sustainability of the Ovine Sector: Eimeria spp. Infections and Nitrogen and Phosphorus Excretions in Dairy Sheep in Italy.

In sheep farming, gastrointestinal parasites can be responsible for significant reductions in animal health and production. Nitrogen (N) and phosphorus (P) fecal excretions are the main determining factors for N2O emissions from manure management and may pose other environmental problems, such as the acidification and eutrophication of natural habitats. By using the Mini-FLOTAC technique on fecal samples from sheep of different ages and physiological status from 19 dairy sheep farms in Tuscany (central Italy), gastrointestinal parasite infections were evaluated. The animal N and P fecal contents were also assessed, with the aim of evaluating possible relationships between the identified parasites and the environmental sustainability of the examined farms. The obtained results showed that Eimeria spp. (86.36%) and gastrointestinal strongyle (54.55%) infections are prevalent in the examined farms. Moreover, significantly higher (p ≤ 0.05) P and Eimeria oocyst/gram-of-feces (OPG) values were found in fecal samples from animals < 1 year of age, and a significant (p ≤ 0.05) positive correlation resulted between N content and Eimeria OPG in fecal samples from animals in the first month of lactation. The findings from this study suggest for the first time that Eimeria spp. infections may have an impact on the environmental sustainability of sheep farming.

Pathomorphological Findings and Infectious Diseases in Selected European Brown Hare (Lepus europaeus Pallas, 1778) Populations from Schleswig-Holstein, Germany.

In the northernmost German federal state Schleswig-Holstein, populations of European brown hares (Lepus europaeus Pallas, 1778) show diverse densities and varying courses over the years. To examine differences in pathomorphological findings and infectious diseases as possible reasons for varying population dynamics, we assessed 155 hunted hares from three locations in Schleswig-Holstein from 2016 to 2020. We investigated the association of location, year, age, and sex of animals to certain pathomorphological findings and infectious diseases. Frequent pathomorphological findings were intestinal parasites (63.9%), hepatitis (55.5%), nephritis (31.0%), steatitis (23.2%), enteritis (13.5%), and pneumonia (5.2%). Body condition differed significantly between locations, and the prevalence of pneumonia was significantly higher in females. Enteritis was not detected in 2019, when much more juveniles were sampled. Hepatitis and nephritis occurred significantly more often in 2016 and among adults. Additionally, more adults showed hepatitis with concurrent serotitre for European brown hare syndrome virus (EBHSV), while intestinal parasitosis as well as high excretion rates of coccidia were more common in juveniles. Sampled animals showed high infection rates with Eimeria spp. (96.1%), Trichostrongylus spp. (52.0%), Graphidium strigosum (41.2%), and a high seroprevalence (90.9%) for EBHSV, without severe symptoms. This study revealed a low prevalence of infectious pathogens, but a high prevalence of chronic inflammations of unknown origin in the tested brown hare populations. Overall, our results indicate a rather minor importance of infectious diseases for observed population dynamics of analysed hare populations in Schleswig-Holstein.

MOLECULAR CHARACTERIZATION OF ORNITHOSTRONGYLUS QUADRIRADIATUS ISOLATED FROM A RACING PIGEON (COLUMBA LIVIA DOMESTICA).

An infestation of Ornithostrongylus quadriradiatus, has been found in a racing pigeon Columba livia domestica). In addition, this individual had a mild infection with Trichomonas spp. and Eimeria spp. The bird showed symptoms of increased appetite; unformed, dark- green feces; and lack of typical male behavior, but the general condition of this pigeon was good. The pigeon was euthanized because of a permanent inability to fly due to an injury to the wing. A postmortem examination revealed the presence of very small dark-pink twisted nematodes with the morphology of O. quadriradiatus in the small intestine, mainly the duodenum. Histopathology showed segmental inflammatory changes in the intestines caused by both blood-sucking nematodes and Eimeria spp. The 5.8S ribosomal ribonucleic acid polymerase chain reaction amplicons from deoxyribonucleic acid extracted from O. quadriradiatus had 82.40% identity with Australostrongylus victoriensis and 81.20% with Travassostrongylus spp. This is the first molecular characterization of O. quadriradiatus-a relatively rare pigeon parasite.

Genetic and phenotypic parameters for sexual precocity and parasite resistance traits in Nellore cattle.

Indicator traits of sexual precocity are widely used as selection criteria for the genetic improvement of beef cattle; however, the impact of selection for these traits on resistance to endoparasites and ectoparasites is unknown. Therefore, this study aimed to estimate the genetic and phenotypic parameters for indicator traits of sexual precocity and parasite resistance in Nellore cattle. The sexual precocity traits evaluated were probability of first calving (PFC) and scrotal circumference at 12 and 18 months of age (SC12 and SC18). The resistance-related traits included tick (TC), gastrointestinal nematode egg (NEC), and Eimeria spp. oocyst (EOC) counts. (Co)variance components were estimated by Bayesian inference using multitrait animal models. The mean heritabilities for PFC, SC12, SC18, TC, NEC, and EOC were 0.23, 0.38, 0.42, 0.14, 0.16, and 0.06, respectively, and suggest that selection will change the mean values of these traits over time. The genetic and phenotypic correlations for most pairs formed by a precocity and a resistance trait were not different from zero, suggesting that selection for sexual precocity traits will not result in changes in resistance traits. Thus, selection for indicator traits of sexual precocity does not elicit unfavorable correlated responses in resistance to endoparasites and ectoparasites, and joint selection aimed at improving these traits can be performed using multitrait selection methods, when necessary.

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.

Effects of supplementing a direct-fed microbial containing Enterococcus faecium 669 on performance, health, and metabolic responses of preweaning Holstein dairy calves.

This study aimed to evaluate the effects of Enterococcus faecium 669 supplementation on performance, health, parasitological, microbiological, and hematological responses of preweaning dairy calves. Forty-two newborn Holstein female calves (initial body weight [BW] 44 ± 4.5 kg) were used in the present study. At birth, calves were ranked by initial BW and assigned to 1 of 2 treatment groups: (1) whole milk (CON; n = 21) and (2) whole milk with the addition of direct-fed microbial Ent. faecium 669 (DFM; n = 21). During the entire experimental period (63 d), DFM was daily-fed at a rate of 2.5 × 109 cfu/head. All calves were offered a mixture of a starter feed and wheat straw for ad libitum consumption. Supplement intake was evaluated daily, whereas calves were weighed on a weekly basis from d 0 to weaning (d 63). Diarrhea was assessed once a day, and fecal and blood samples were collected for microbiological, parasitological, and hematological responses. All data were analyzed with SAS using calf as the experimental unit. A treatment × week interaction was observed for BW, as DFM-supplemented calves were heavier than CON cohorts on d 56 (+ 4.7 kg) and at weaning on d 63 (+ 4.8 kg). A similar interaction was observed for average daily gain (ADG) and dry matter intake (DMI), with greater ADG for DFM-supplemented calves from d 35 to 42, greater ADG and DMI from d 49 to 56, and greater DMI from d 56 to weaning. Moreover, diarrhea occurrence tended to be lower, whereas rectal temperature was 0.2°C lower for DFM-supplemented calves. Treatment × day interactions were observed for the occurrence and counts of Eimeria spp., as DFM-supplemented calves tended to have a reduced number of positive observations on d 42 of the study versus CON, and a significant reduction in positive animals from d 21 to 42 was observed in the DFM group but not in CON calves. For Cryptosporidium spp., no treatment effects were observed on overall occurrence (%), but DFM-supplemented calves had a greater count of oocyst per gram versus CON. No treatment × day interaction or main treatment effects were observed for any of the blood variables analyzed herein, exception being monocytes concentration. In summary, preweaning Ent. faecium 669 supplementation improved performance, diarrhea occurrence, and reduced the number of calves positively-detected for Eimeria spp.

First report on the molecular prevalence and associated risk factors of Eimeria spp. in dairy cattle in Khon Kaen, Thailand.

Background and Aim: Bovine coccidiosis, caused by the protozoa Eimeria, is an important parasitic cattle disease that affects animal health and has economic impact worldwide. This study was conducted to report the first molecular prevalence and genetic diversity of Eimeria spp. in dairy cattle in Khon Kaen province, Thailand, and to identify the risk factors associated with Eimeria spp. infection. Materials and Methods: From July 2020 to October 2021, 296 fecal samples were collected from dairy cattle divided into three age groups, including <3-month-old calves, 3-month-old to 1-year-old calves, and >1-year-old cattle. Eimeria spp. were identified by multiplex polymerase chain reaction (PCR) amplifying 18S RNA gene and confirmed by DNA sequencing. Information regarding all associated risk factors was collected using questionnaires and analyzed using logistic regression tests in the Statistical Package for the Social Sciences program. Results: Polymerase chain reaction results showed that 104 (35.13%) of 296 samples were positive for Eimeria spp. The <3-month-old calves (46.51%) had the highest infection rate. Moreover, multiplex PCR identified five species of Eimeria, namely, Eimeria bovis (32.69%), Eimeria zuernii (18.26%), Eimeria alabamensis (5.76%), Eimeria ellipsoidalis (3.84%), and Eimeria cylindrica (2.88%). An association was observed between risk factors and Eimeria spp. incidence (p < 0.05). DNA sequencing revealed the similarity of each Eimeria spp. with 91%-100% nucleotide identity. Phylogenetic tree analysis demonstrated the close relationships of clusters of E. bovis and E. zuernii, E. ellipsoidalis, and E. cylindrica and another cluster of E. alabamensis. Conclusion: The results confirm that Eimeria spp. are commonly found in dairy cattle, especially calves. The molecular test could be powerful for species identification. This study also provides epidemiological information for developing future strategies to control bovine coccidiosis.

Implementing real-time immunometabolic assays and immune cell profiling to evaluate systemic immune response variations to Eimeria challenge in three novel layer genetic lines.

Introduction: Evaluating differences in immune responses to Eimeria spp. between poultry genetic lines could be valuable for understanding favorable traits to address coccidiosis, a costly poultry disease. The objective was to compare peripheral blood mononuclear cell (PBMC) immunometabolism and composition during Eimeria challenge in three distinct and highly inbred genetic lines; Leghorn Ghs6, Leghorn Ghs13, and Fayoumi M5.1. Methods: At hatch, 180 chicks (60/ line) were placed in wire-floor cages (10 chicks/cage) and fed a commercial diet. Baseline PBMC were isolated on d21 (10 chicks/line) and 25 chicks/line were inoculated with 10X Merck CocciVac®-B52 (Kenilworth, NJ), creating 6 genetic line × Eimeria groups total. Chicks were euthanized on 1, 3, 7, and 10d post-inoculation (pi; 5 chicks/ line × Eimeria group) for PBMC isolation with body weight and feed intake recorded throughout. Immunometabolic assays to determine PBMC ATP production profiles and glycolytic activity were implemented along with flow cytometric immune cell profiling. Genetic line × Eimeria challenge, and line´challenge fixed effects were analyzed using the MIXED procedure (SAS 9.4; P ≤ 0.05). Results and Discussion: Before inoculation, M5.1 chicks had 14.4-25.4% greater average daily gain (ADG) with 19.0-63.6% increased monocyte/macrophage+, Bu-1+ B cell, and CD3+ T cell populations compared to both Ghs lines (P < 0.0001) but similar immunometabolic phenotype. The Eimeria main effect reduced ADG by 61.3% from 3-7dpi (P = 0.009) except in M5.1 chicks, where no ADG difference due to challenge was found. At 3dpi, Eimeria-challenged M5.1 chicks had 28.9 and 33.2% reduced PBMC CD3+ T cells and CD3+CD8α+ cytotoxic T cells than unchallenged chicks, suggesting early and preferential recruitment from systemic circulation to tissues local to Eimeria challenge (i.e., intestine; P ≤ 0.01). Both Ghs lines displayed 46.4-49.8% T cell reductions at 10dpi with 16.5-58.9% recruitment favoring underlying CD3+CD4+ helper T cells. Immunometabolic responses in Eimeria-challenged Ghs6 and Ghs13 chicks were characterized by a 24.0-31.8% greater proportion of ATP from glycolysis compared to unchallenged counterparts at 10dpi (P = 0.04). These results suggest that variable T cell subtype recruitment timelines in addition to altered systemic immunometabolic requirements may work synergistically to determine favorable immune responses to Eimeria challenge.

Finisher lamb growth and rumen fermentation responses to the plane of nutrition and naturally occurring coccidiosis.

The objective of the present study was to investigate the interaction of plane of nutrition and naturally occurring coccidiosis on finisher lamb growth performance, FAMACHA score, and rumen volatile fatty acid profile. The study included 30 Suffolk, Dorset or Suffolk x Dorset lambs and were divided into 2 groups based on their initial body weight and assigned to 2 feeding groups differing in dietary energy intake to create lambs representing divergent growth curves due to differing nutritional management. Lambs with naturally occurring coccidiosis and healthy lambs were present in both feeding groups making a 2 × 2 factorial arrangement of treatments, (a) high plane of nutrition (HPN) lambs with no clinical coccidiosis diagnosis (HPNH), (b) HPN lambs with clinical coccidiosis (HPNC), (c) low plane of nutrition (LPN) lambs with no clinical coccidiosis diagnosis (LPNH), (d) LPN lambs with clinical coccidiosis (LPNC). Body weight and FAMACHA scores were recorded once every 2 weeks. On d 65 of feeding, lambs were slaughtered, and rumen fluid samples were collected and analyzed for volatile fatty acid concentrations. All response variables were analyzed statistically using a linear mixed effects model with fixed effects for plane of nutrition, health status, and a random effect for initial body weight nested within the pen. The total and average weight gain were not associated with planes of nutrition, health status, or the interaction. Health status had an impact on FAMACHA© score (P = 0.047) and concentration of isobutyrate (P = 0.037) and tended to affect total VFA (P = 0.085) and acetate (P = 0.071) concentrations. The interaction between the plane of nutrition and the health status tended to affect butyrate concentration (P = 0.058). These data support the conclusion that coccidiosis infection impacted on rumen fermentation in a manner independent of the plane of nutrition; however, the translation of these rumen level impacts did not translate to the production responses.

Novel Antioxidant Insights of Myricetin on the Performance of Broiler Chickens and Alleviating Experimental Infection with Eimeria spp.: Crosstalk between Oxidative Stress and Inflammation.

In the modern poultry industry, the application of novel phytogenic bioactive compounds with antioxidant potential aims to enhance productivity and quality and to minimize the stress of associated diseases. Herein, myricetin, a natural flavonoid, was evaluated for the first time on broiler chickens' performance, antioxidants and immune modulating functions, and tackling avian coccidiosis. A total of 500 one-day-old chicks were divided into five groups. The negative (NC) and infected control (IC) groups were fed a control diet without additives, and the latter was infected with Eimeria spp. Groups supplemented with myricetin (Myc) were fed a control diet of Myc (200, 400 and 600 mg/kg diet each). On d 14, all chicks except those in NC were challenged with oocysts of mixed Eimeria spp. Significant improvements in the overall growth rate and feed conversion ratio were detected in the group that was fed 600 mg/kg, unlike the IC group. Notably, groups that were fed 400 and 600 mg/kg showed higher total meat antioxidant capacity with an inverse reduction in oxidative and lipid peroxidation biomarkers (hydrogen peroxide: H2O2; reactive oxygen species: ROS; Malondialdehyde: MDA). Of note, the upregulation of glutathione peroxidase; GSH-Px, catalase; CAT, superoxide dismutase; SOD, heme oxygenase-1; HO-1 and NAD(P)H dehydrogenase quinone 1 NQO1 genes in jejunum and muscle were prominently observed with increasing levels of supplemental Myc. At 21 dpi, the severity of coccoidal lesions (p < 0.05) induced by mixed Eimeria spp. and oocyst excretion were greatly reduced in the group that was fed 600 mg/kg of Myc. In the IC group, higher serum levels of C-reactive protein; CRP and nitric oxide; and NO and the upregulated expression of inflammatory biomarkers (interleukin-1ß; IL-1ß, interleukin-6; IL-6, tumor necrosis factor-α; TNF-α, chemotactic cytokines; CCL20, stromal cell-derived factor-1; CXCL13, and avian defensins; AvBD612) were subsided in higher levels in the Myc-fed groups. Taken together, these findings indicate the promising antioxidant role of Myc in modulating immune responses and reducing growth depression associated with coccidia challenges.

Molecular Investigation of Eimeria Species in Broiler Farms in the Province of Vojvodina, Serbia.

Coccidiosis is a significant poultry disease caused by the Eimeria species. This study aims to determine the prevalence of Eimeria spp. on broiler farms in Vojvodina, along with the identification of parasite species, and assess the implemented biosecurity measures. The study was conducted on 100 broiler chicken farms (28 small-sized; 34 medium-sized; 38 large-sized farms) from June 2018 to December 2021. One pooled sample of faeces was collected from three to six-week-old chickens from each farm, and assessment of biosecurity measures was carried out using a questionnaire. Using the PCR method, DNA of Eimeria was found in 59 samples (59%), while 41 samples (41%) were negative. Four species of Eimeria were identified, and their prevalence was the following: E. acervulina (37%), E. maxima (17%), E. mitis (25%) and E. tenella (48%). A significant difference (p < 0.05) was established in the number of oocysts in flocks from small-sized farms compared to medium-sized farms. It was found that regular implementation of disinfection, disinsection and deratisation measures, as well as all the biosecurity measures, can significantly reduce the occurrence of coccidiosis. These results will help to develop better strategies for the control and prevention of coccidiosis on farms.

Gastrointestinal Parasites of Domestic Mammalian Hosts in Southeastern Iran.

Gastrointestinal parasites (GIP) are a major cause of disease and production loss in livestock. Some have zoonotic potential, so production animals can be a source of human infections. We describe the prevalence of GIP in domestic mammals in Southeastern Iran. Fresh fecal samples (n = 200) collected from cattle (n = 88), sheep (n = 50), goats (n = 23), camels (n = 30), donkeys (n = 5), horse (n = 1), and dogs (n = 3) were subjected to conventional coprological examination for the detection of protozoan (oo)cysts and helminth ova. Overall, 83% (166/200) of the samples were positive for one or more GIP. Helminths were found in dogs, donkeys, sheep (42%), camels (37%), goats (30%), and cattle (19%), but not in the horse. Protozoa were found in cattle (82%), goats (78%), sheep (60%), and camels (13%), but not in donkeys, dogs, or the horse. Lambs were 3.5 times more likely to be infected by protozoa than sheep (OR = 3.5, 95% CI: 1.05-11.66), whereas sheep were at higher odds of being infected by helminths than lambs (OR = 4.09, 95% CI: 1.06-16.59). This is the first study assessing the prevalence of GIP in domestic mammals in Southeastern Iran.

Prevalence, coinfection, and risk factors associated with Fasciola hepatica and other gastrointestinal parasites in cattle from the Peruvian Amazon.

Background and Aim: Extensive cattle rearing is a major source of economy for the inhabitants of the Amazon region of Peru. Milk and meat production is generally affected by the prevalence of various parasites, including hepatic and gastrointestinal parasites, as these products provide ideal conditions for parasitic growth. This poses a serious public health threat. This study aimed to estimate the prevalence, coinfection, and risk factors associated with the liver fluke (Fasciola hepatica) and other gastrointestinal parasites in cattle from the Amazon region of Peru. Materials and Methods: Fecal samples obtained from 1450 bovine specimens were analyzed using flotation and sedimentation methods to identify parasites, including Eimeria spp., strongyle-type eggs (STEs), and F. hepatica. We collected information about the specimens, including age, sex, origin, breed, category, frequency of deworming, farm size, herd size, water sources, and rearing system by conducting simple inspections and interviewing owners. The data obtained were statistically evaluated using the Chi-square test (p < 0.05) to determine the association between the qualitative variables. We also calculated the odds ratio at a 95% confidence interval to identify the risk factors. Results: We observed that F. hepatica, Eimeria spp., and STEs were 45.6%, 39.8%, and 35.3% prevalent, respectively. We found risk factors related to distomatosis in the animals from Huambo, where the drinking water sources are mainly streams, ditches, and rivers, while the specimens from Valle Chico were predisposed to coccidiosis. Further, the risk factors related to the presence of STEs in feces were age (61-90 months), origin (Valle Chico), herd size (<50 animals), and type of extensive rearing. Furthermore, significant coinfection was observed between Eimeria spp. and STEs. Conclusion: The high percentages of parasites in cattle observed were related to epidemiological factors, such as the origin of the sample, water sources, age, herd size, and extensive breeding. Similarly, the presence of STEs was a risk factor for contracting coccidiosis. Our future goals include investigating these parasites using a larger sample size and identifying more risk factors using more sensitive and specific diagnostic tests.

Effect of Supplementation of Lambs with Whole Cottonseed: Impact on Serum Biomarkers and Infection by Gastrointestinal Parasites under Field Conditions.

The purpose of this trial was to evaluate serum levels of oxidative stress biomarkers and biochemical analytes in crossbred lambs during the rearing phase in an integrated crop-livestock system (ICLS) to control gastrointestinal parasites. The experiment used 36 crossbred lambs (cross: Ile de France × White Dorper × Texel) divided into two groups. The WCS group was supplemented with whole cottonseed (WCS), and controls had no supplementation. Body weight, blood collection, and fecal analysis of nematode eggs and Eimeria oocysts counting per gram of feces were performed for each animal within 84 days of experiment. The following serum analytes were determined: total protein, albumin, globulin, cholesterol, haptoglobin, and 10 oxidative stress biomarkers: cupric reducing antioxidant capacity, ferric reducing ability of plasma, trolox equivalent antioxidant capacity, thiol, uric acid, paraoxonase-1, total oxidant status, ferric-xylenol orange, advanced oxidation protein products, and reactive oxygen metabolites derived compounds. The inclusion of WCS suggested the benefit in controlling infection as well as inducing an increase in antioxidants and a decrease in oxidants in lambs naturally infected by gastrointestinal parasites. The combination of WCS and ICLS could be a useful tool in controlling gastrointestinal parasite infection without affecting the production performance.