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Eimeria bovis is a pathogenic protozoan that causes cattle digestive tract infections, which can cause economic losses to farmers. It is necessary to develop specific and accurate detection methods to conserve livestock and prevent coccidiosis in Indonesia. This study aims to detect E. bovis by nested PCR and determine the relationship with reference sequences. A total of 167 samples of beef cattle feces were taken randomly from community farms spread across 18 provinces in Indonesia. The feces were examined natively, and then the oocysts were purified by the sugar flotation method, extracted by KIT extraction, and amplified by the nPCR technique. Positive samples were followed by sequencing and phylogenetic analysis using MEGA 11 software. This study used two pairs of primers (outer and inner) taken from ITS-1 molecular markers. As many as 96 out of 167 samples (57.5%) were positive for Eimeria spp., and 48 of the 96 samples were positive for Eimeria spp. (50%) were detected to be positive for E. bovis based on the presence of a 238 bp DNA fragment. The results of the phylogenetic analysis showed that the study sample formed a separate cluster from the E. bovis cluster from abroad. In conclusion, E. bovis was detected in 16 out of 18 provinces in this study, and the nPCR technique proved to have better sensitivity and specificity. [ABSTRACT FROM AUTHOR]

1. Two experiments were conducted, the first was to investigate the effect of increasing digesta viscosity by dietary carboxymethyl cellulose (CMC) on the growth performance and intestinal morphology and characteristics of healthy birds. The second experiment evaluated the impact of increased digesta viscosity in birds during an Eimeria spp. challenge.2. In experiment 1, a corn-soybean meal-based basal diet was supplemented with 0, 10 or 20 g/kg CMC at the expense of cornstarch and offered to seven birds in each of eight replicate cages per diet from d 8 to 22 post hatching.3. Increasing digesta viscosity due to dietary CMC linearly reduced (p < 0.05) body weight (BW) gain and the apparent ileal digestibility of nutrients. The relative lengths of the duodenum, jejunum and ileum linearly increased (p < 0.01) with dietary CMC inclusion.4. In experiment 2, on d 14, 256 broiler chickens were randomly assigned to eight replicate cages in a 2 × 2 factorial arrangement of treatments with two CMC concentrations (0 or 10 g CMC/kg of diet), with or without an Eimeria challenge. On d 15, birds in the challenge groups were orally gavaged with a 1 ml solution containing 25,000, 25,000 or 125,000 oocysts of E. maxima, E. tenella and E. acervulina; or 1% PBS, respectively.5. Increasing digesta viscosity in Eimeria-challenged birds decreased the total tract digestibility of dry matter and gross energy (p < 0.05). The ileal gene expression of glucose transporters was upregulated (p < 0.05) in challenged birds that received the CMC-supplemented diet.6. In summary, increased digesta viscosity induced changes in the expression of nutrient transporter genes and decreased nutrient utilisation in Eimeria-challenged birds. [ABSTRACT FROM AUTHOR]

Anticoccidial vaccines comprising living oocysts of Eimeria tenella, Eimeria necatrix, Eimeria maxima, and Eimeria acervulina are used to control coccidiosis. This study explored the potential of IL-1β to act as a molecular adjuvant for enhancing the immunogenicity of Eimeria necatrix and mucosal immunity. We engineered E. necatrix to express a functional chIL-1β (EnIL-1b) and immunized chickens with oocysts of the wild type (EnWT) and tranegenic (EnIL-1b) strains, respectively. The chickens were then challenged with EnWT oocysts to examine the immunogenicity-enhancing potential of chIL-1b. As expected, the oocyst output of EnIL-1β-immunized chickens was significantly reduced compared to those immunized using EnWT. No difference in body weight gain and lesion scores of EnIL-1β and EnWT groups was observed. The parasite load in the small intestine and caeca showed that the invasion and replication of EnIL-1β was not affected. However, the markers of immunogenicity and mucosal barrier, Claudin-1 and avian β-defensin-1, were elevated in EnIL-1binfected chickens. Ectopic expression of chIL-1β in E. necatrix thus appears to improve its immunogenicity and mucosal immunity, without increasing pathogenicity. Our findings support chIL-1β as a candidate for development of effective live-oocyst-based anticoccidial vaccines. [ABSTRACT FROM AUTHOR]

Birds of various families and genera comprise a notable proportion of veterinary clients in veterinary practice. Regarding the scarcity of information on avian coccidia in pet birds, this study was designed to investigate and identify the diversity of gastrointestinal coccidian protozoa in caged pet birds. Accordingly, droppings samples were collected in 2.5% potassium dichromate and examined for the presence of oocysts by wet smear and by flotation with the saturated salt solution. Detected oocysts were further morphologically specified. Special Modified Ziehl-Neelsen stain was used for the detection of Cryptosporidium spp. oocysts. Out of the 145 droppings that were collected for parasitological investigations, it was discovered that a Japanese quail (Coturnix japonica) had a Cryptosporidium sp. infection (1/145: 0.7%). Additionally, Isospora spp. was found in a common mynah and a canary (2/145: 1.4%), while Eimeria sp. was detected in a canary (1/145: 0.7%). Studied birds were clinically normal and no signs of intestinal infection were observed. The oocysts are described morphologically in detail and compared with the available data. Periodic faecal examination should be recommended for monitoring intestinal parasites in pet birds both from a public health perspective for Cryptosporidium and for avian host health in the case of non-zoonotic parasites. Additionally, the identification of parasite/host species composition will enhance our understanding of species diversity and the variety of hosts that coccidian parasites are capable of infecting. [ABSTRACT FROM AUTHOR]

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. This study aimed to evaluate the effects of direct-fed microbials (DFM) on health and growth responses of preweaning Bos indicus × Bos taurus (Gyr × Holstein) crossbred calves. Ninety newborn heifer calves (initial BW of 35 ± 4.0 kg) were used. At birth, calves were ranked by initial BW and parity of the dam and assigned to: (1) whole milk without DFM supplementation (CON; n = 30), (2) whole milk with the addition of 1.0 g/calf per day of a Bacillus -based DFM (BAC; n = 30), or (3) whole milk with the addition of 1.0 g/calf per day of BAC and 1.2 g/calf per day of Enterococcus faecium 669 (MIX; n = 30). Milk was fed individually during the study (77 d), and the BAC and MIX treatments were offered daily throughout the 77-d preweaning period. All calves were offered a starter supplement and corn silage starting on d 1 and 60 of age, respectively. Milk and starter supplement intake were evaluated daily, and BW was recorded on d 0 and at weaning (d 77). Diarrhea and pneumonia were assessed daily, and fecal samples were collected on d 0, 7, 14, 21, and at weaning (d 77) for assessment of the presence of bacterial and protozoal pathogens via qPCR. All data were analyzed using SAS (v. 9.4) with calf as the experimental unit and using single-df orthogonal contrasts (BAC + MIX vs. CON; BAC vs. MIX). Daily feeding of DFM, regardless of type, improved weaning BW. Odds ratio for occurrence of pneumonia was lower for DFM-supplemented calves, but its occurrence did not differ between BAC and MIX calves. No Salmonella spp. or Escherichia coli F41 were detected in any of the calves. The proportion of calves positive for E. coli F17 was greater for DFM calves on d 7 (92% and 96% vs. 81% for BAC, MIX, and CON, respectively), on d 21 (13% and 26% vs. 7% for BAC, MIX, and CON, respectively), and at weaning (48% and 35% vs. 22% for BAC, MIX, and CON, respectively). For Clostridium difficile , more DFM calves were positive on d 7 (65% and 30% vs. 35% for BAC, MIX, and CON, respectively) and 14 (20% and 28% vs. 7% for BAC, MIX, and CON, respectively), but proportion of positive calves was also greater for BAC versus MIX on d 7. More CON calves were positive for Clostridium perfringens on d 14 (14% vs. 3% and 8% for CON, BAC, and MIX, respectively) compared with DFM-fed calves. Incidence of calves positive for C. perfringens was greater in BAC than MIX on d 7 (50% vs. 18%), and greater for MIX than BAC at weaning (9% vs. 0%). For protozoa occurrence, a lower proportion of DFM calves were positive for Cryptosporidium spp. on d 7 (58% and 48% vs. 76% for BAC, MIX, and CON, respectively), but opposite results were observed on d 21 for Cryptosporidium spp. (3% and 11% vs. 0% for BAC, MIX, and CON, respectively) and Eimeria spp. on d 14 (7% and 8% vs. 0% for BAC, MIX, and CON, respectively) and 21 (50% and 59% vs. 38% for BAC, MIX, and CON, respectively). In summary, DFM feeding alleviated the occurrence of pneumonia and improved growth rates, while also modulating the prevalence of bacteria and protozoa in preweaning Gyr × Holstein calves. [ABSTRACT FROM AUTHOR]

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of diclazuril (Clinacox® 0.5%) as a coccidiostat for chickens for fattening and chickens reared for laying. The additive currently on the market complies with the existing conditions of authorisation. The additive remains safe for the target species and the consumer under the authorised conditions of use. The additive is irritant to skin, eyes and respiratory tract but is not a skin sensitiser. Exposure by inhalation cannot be excluded. The FEEDAP Panel cannot conclude on the safety for the environment of diclazuril from Clinacox® 0.5% due to lack of data. Diclazuril from Clinacox® 0.5% at a concentration of 1 mg diclazuril/kg complete feed has the potential to control coccidiosis in chickens for fattening. This conclusion is extended to chickens reared for laying. Development of resistance to diclazuril of field Eimeria spp. strains isolated from chickens should be monitored. [ABSTRACT FROM AUTHOR]

The 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 HIGHLIGHTS Cubic 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. [ABSTRACT FROM AUTHOR]

The codon usage bias (CUB) of genes encoded by different species' genomes varies greatly. The analysis of codon usage patterns enriches our comprehension of genetic and evolutionary characteristics across diverse species. In this study, we performed a genome-wide analysis of CUB and its influencing factors in six sequenced Eimeria species that cause coccidiosis in poultry: Eimeria acervulina, Eimeria necatrix, Eimeria brunetti, Eimeria tenella, Eimeria praecox, and Eimeria maxima. The GC content of protein-coding genes varies between 52.67% and 58.24% among the six Eimeria species. The distribution trend of GC content at different codon positions follows GC1 > GC3 > GC2. Most high-frequency codons tend to end with C/G, except in E. maxima. Additionally, there is a positive correlation between GC3 content and GC3s/C3s, but a significantly negative correlation with A3s. Analysis of the ENC-Plot, neutrality plot, and PR2-bias plot suggests that selection pressure has a stronger influence than mutational pressure on CUB in the six Eimeria genomes. Finally, we identified from 11 to 15 optimal codons, with GCA, CAG, and AGC being the most commonly used optimal codons across these species. This study offers a thorough exploration of the relationships between CUB and selection pressures within the protein-coding genes of Eimeria species. Genetic evolution in these species appears to be influenced by mutations and selection pressures. Additionally, the findings shed light on unique characteristics and evolutionary traits specific to the six Eimeria species. [ABSTRACT FROM AUTHOR]

Background: Over the last decade, extensive use of coccidiostats to treat and control Eimeria infection has developed drug resistance, prompting the search for new alternative therapies. Rhatany is proven to have various pharmacological properties. Objective: The present study aimed to in vitro and in vivo evaluate the effect of Rhatany roots extract (RRE) as an anti-eimerial and anti-apoptotic agent against murine eimeriosis induced by Eimeria papillata. Methods: Phytochemical screening by gas chromatography-mass spectrometry analysis (GC-MS) was used to detect active compounds in RRE. In vitro anti-eimerial activity of RRE (200, 100, 50 mg/ml), amprolium, phenol, Dettol™, and formalin were studied after incubation with non-sporulated Eimeria oocysts. For the in vivo study, twenty-five male C57BL/6 mice were randomly allocated into five groups. Animals in the first group were just given distilled H2O, while those in the second group were given 200 mg/kg RRE for 5 days. The Eimeria parasite’s oocysts were infected into the third, fourth, and fifth groups. For treatment, RRE (200 mg/kg) and amprolium (120 mg/kg) were orally given to the 4th and 5th groups for five days, respectively. All mice were euthanized, on day 5 post-infection, to collect the jejunal tissues under study. Investigations were undertaken into the oocyst output in feces and goblet cells in mice jejuna. Assays for glutathione peroxidase (GPx), hydrogen peroxide (H2O2), and myeloperoxidase (MPO) were also performed. In jejunal tissue, cysteine aspartic acid protease-3 (Caspase-3) was counted using immunohistochemistry, while BCL2-associated X protein (Bax) and B-cell lymphoma-2 (BCL-2) were assayed using ELISA. In addition, mRNA expression of the goblet cell response gene (MUC2) was detected using real-time PCR. Results: Phytochemical screening by GC-MS demonstrated the presence of 22 compounds in the RRE. The in vitro study revealed that RRE significantly inhabited the oocyst sporulation in a dose-dependent manner. By day 5 after infection with the Eimeria parasite, the number of oocysts in mice feces was significantly reduced after RRE treatment (1.308 × 106 ± 1.36 × 105 oocysts/g feces) compared to the infected group (5.387 × 106 ± 4.29 × 105 oocysts/g feces). Moreover, the Eimeria infection reduced the number of goblet cells of mice jejuna and its specific gene, MUC2. The treatment with RRE increased the number of goblet cells/villus from 3.45 ± 0.17 to 6.04 ± 0.23, associated with upregulation for MUC2 from 0.26 to 2.39-fold. Also, the Eimeria experimental infection lowered the activity of the antioxidant enzyme represented by GPx (23.99 ± 3.68 mg/g tissue), while increasing the stress parameters of hydrogen peroxide (0.07 ± 0.01 mM/g) as well as the activity of MPO (66.30 ± 3.74 U/mg). The production of apoptotic markers including Caspase-3 (68.89 ± 2.67 U/g) and Bax (159.05 ± 6.50 pg/ml) was significantly elevated while decreasing the anti-apoptotic marker of BCL2 (0.42 ± 0.07 pg/ml). Our study proved that RRE significantly reduced oxidative stress, and apoptotic markers as well as the inflammatory activity of MPO. Also, antioxidant enzyme and anti-apoptotic activity in the jejunum of E. papillata-infected mice were enhanced after RRE treatment. Conclusion: Our study highlights the potential of RRE as a natural solution for coccidiosis management by modulating apoptosis in E. papillata host cells. However, further research is needed to fully understand the underlying mechanisms and enhance our understanding of its therapeutic efficacy. [ABSTRACT FROM AUTHOR]

Coccidiosis is one of the most prevalent diseases found in local rabbits (Oryctolagus cuniculus), which is caused by the Eimeria. The study aimed to more reliably identify Eimeria species (Eimeria magna) infecting Local Rabbits in Alkarg City, Saudi Arabia, based the method on the molecular properties and morphological and molecular biological techniques. Sub‐spheroidal oocysts measuring 21–27 × 12–16 (24 × 14.4) μm (20 n) and with a length/width (L/W) ratio of 0.9–1.1 (1.0) were identified by microscopic analysis of a fecal sample. Oocysts feature a bi‐layered wall that is 1.0–1.2 (1.1) μm thick. About two‐thirds of the wall's thickness is made up of a smooth outer layer. A polar granule is present, but neither a micropyle nor an oocyst residuum is present. The ovoidal sporozoites measure 15–18 × 8–11 (16.5 × 9.5) μm, have an L/W ratio of 1.6–1.8 (1.7), and take up around 21% of the oocyst's total surface. The mean size of the sub‐Stieda body is 1.4 × 2.3 μm, while the average size of the Stieda body is 0.9 × 1.8 μm. The para‐Stieda body is lacking. Sporocyst residuum appears membrane‐bound and has an uneven form made up of several granules. With two refractile bodies below the striations and pronounced striations at the more pointed end, sporozoites are vermiform, measuring an average of 11.6 × 4.0 μm. The results of the sequencing for the 18S rDNA gene confirmed the species of Eimeria parasites found in the host (rabbits). The current parasite species is closely related to the previously described and deposited E. magna and deeply embedded in the genus Eimeria (family Eimeriidae). According to the findings, single oocyst molecular identification of Eimeria may be accomplished through consistent use of the morphological and molecular results. It is possible to draw the conclusion that the current research supplies relevant facts that help assess the potential infection and future control measures against rabbit coccidiosis to reduce the financial losses that can be incurred by the rabbit industry in Saudi Arabia. [ABSTRACT FROM AUTHOR]

Background: Eimeria is a protozoan parasite that affects poultry, particularly chickens, causing a disease known as coccidiosis. This disease imposes substantial significant economic challenges to the poultry sector. Objectives: The current study aimed to estimate the global prevalence and associated risk factors of Eimeria in domestic chickens. Methods: Multiple databases (Scopus, PubMed, ProQuest, Web of Science and Google Scholar) were searched for articles published until June 2023. The pooled prevalence was estimated using a random‐effects model with a 95% confidence interval. The statistical analysis was conducted using meta packages in R version (3.6.1). Results: In total, 41 articles fulfilled the eligibility criteria. The global pooled prevalence was 44.3% (36.9%–51.8%) with Eimeria tenella (38.7%, 30.1%–47.7%) as the most prevalent species. The highest pooled prevalence was related to the Western Pacific Region (80.5%, 72.6%–87.3%) and urban areas (44.4%, 36.5%–52.6%). Moreover, areas with humid subtropical climates represent the highest overall prevalence (75.8%, 46.6%–95.9%). Conclusion: The necessity for robust and innovative strategies for preventing and managing this disease cannot be overstated. Addressing Eimeria impact is crucial not only for safeguarding poultry health but also for sustaining the economic viability of the poultry industry. [ABSTRACT FROM AUTHOR]

In poultry rearing on deep bedding systems, microorganisms that cause infectious diseases develop within the bedding. This study aimed to determine the physical properties and microbial accumulation capacity of different types of bedding. Methods used included thermo-programmed desorption mass spectrometry, microbiological methods, and scanning electron microscopy. The hygroscopic properties of straw bedding deteriorated by 269.90% from the seventh to the fortysecond day. In the experimental room, where a dry disinfectant was added to the straw bedding, the hygroscopic capacity was 86.70% at the end of the experiment, a difference of 183.2%. Wood shavings bedding, with the addition of disinfectant, exhibited a water absorption capacity 102.82% higher than the control on the forty-second day. Granulated bedding with disinfectant retained moisture 150.33% better compared to the control. At the end of the experiment, the pH of straw bedding was 8.13 in the control group compared to 7.56 in the treated group; for wood shavings was 7.95 versus 7.16; and for granule was 7.35 versus 6.35, due to the use of the disinfectant. At the conclusion of the experiment, granulated bedding exhibited the lowest pH compared to straw and wood shavings, affecting the growth and reproduction of microorganisms. Monitoring of microorganisms in the bedding revealed the presence of bacteria: Escherichia coli, Salmonella typhimurium, Salmonella pullorum, Clostridium perfringens, Staphylococcus aureus, Klebsiella pneumonia, Listeria monocytogenes, as well as microscopic fungi: Fusarium sporotrichioides, Aspergillus niger, and Eimeria oocysts. The use of disinfectant reduced bacterial contamination of straw bedding by 12.35-199.19%, fungal contamination by 633.91-1959.14%, and Eimeria by 676.50%. In the wood shavings bedding, bacterial contamination was reduced by 32.84-257.93%, fungal contamination by 487.08-1098.4%, and Eimeria by 570.21%. Granulated bedding showed reduced bacterial accumulation by 50.93-228.87%, fungal accumulation by 169.89-500.9%, and Eimeria by 301.56%. The practical significance of this study lies in the improved physical properties of poultry bedding and the reduction in bacterial, fungal, and Eimeria oocyst accumulation

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. [ABSTRACT FROM AUTHOR]

Abstract Background This study investigated effects of different methionine (Met) supplementation levels in a reduced protein diet on growth performance, intestinal health, and different physiological parameters in broilers under Eimeria challenge. A total of 600 fourteen-day-old Cobb500 male broilers were challenged with E. maxima, E. tenella, and E. acervulina, and randomly allocated in a 2 × 5 factorial arrangement. Birds received normal protein diets (20% crude protein, NCP) or reduced protein diets (17% crude protein, LCP), containing 2.8, 4.4, 6.0, 7.6, and 9.2 g/kg of Met. Results On 6 and 9 days post inoculation (DPI), increasing Met level linearly improved the growth performance (P

Abstract Background Coccidiosis is one of the most frequently reported diseases in chickens, causing a significant economic impact on the poultry industry. However, there have been no previous studies evaluating the prevalence of this disease in broiler farms in Guangdong province. Therefore, this study aims to conduct an epidemiological investigation into the occurrence of Eimeria species and associated risk factors in intensive management conditions across four regions in Guangdong province, China. A total of 394 fecal samples were collected from 89 broiler farms in Guangdong province. The prevalence of Eimeria species infection was determined using PCR, and the occurrence of Clostridium perfringens type A was assessed using quantitative real-time PCR. Results The results showed an overall prevalence of 98.88% (88/89) at the farm level and 87.06% (343/394) at the flock level. All seven Eimeria species were identified, with E. acervulina (72.53%; 64/89), E. tenella (68.54%; 61/89), and E. mitis (66.29%; 59/89) at the farm level, and E. acervulina (36.55%; 144/394), E. mitis (35.28%; 139/394), and E. tenella (34.01%; 134/394) at the flock level. The predominant species combination observed was a co-infection of all seven Eimeria species (6.74%; 6/89), followed by a combination of E. acervulina, E. tenella, E. mitis, E. necatrix, E. brunetti, and E. maxima (5.62%, 5/89). A combination of E. acervulina, E. tenella, E. mitis, E. necatrix, E. brunetti, and E. praecox (4.49%; 4/89) was also observed at the farm level. Furthermore, the study identified several potential risk factors associated with the prevalence of Eimeria species, including farm location, chicken age, drinking water source, control strategy, and the presence of C. perfringens type A were identified as potential risk factors associated with prevalence of Eimeria species. Univariate and multivariate analyses revealed a significant association between E. necatrix infection and both grower chickens (OR = 10.86; 95% CI: 1.92–61.36; p

This study aims to determine the prevalence of Cryptosporidium and Eimeria spp. oocysts in fish specimens in the river Kura. It was conducted during the 2021–2022 at two sites: Mingachevir reservoir in central Azerbaijan and in Neftchala district where the river finally enters the Caspian Sea through a delta of the Kura River estuary. The diagnosis of oocysts was performed microscopically. Fine smears from the intestine epithelial layers stained by Ziehl-Neelsen for Cryptosporidium oocysts. To identify Eimeria oocysts, each fish's faecal material and intestinal scrapings were examined directly under a light microscope in wet samples on glass slides with a coverslip. Results revealed a prevalence of Cryptosporidium and Eimeria species infections in fish hosts from both territories Rutilus caspicus, Alburnus filippi, Abramis brama orientalis and Carassius gibelio. Of 170 investigated fish specimens, 8.8% (15/170) were infected with Cryptosporidium species oocysts. Eimeria species oocysts were identified in 20.6% (35/170). The presence of Cryptosporidium and Eimeria infections in fish specimens are natural infections. However, their presence in fish species may be attributed to the age of the fish species and water pollution. This is the first report regarding the prevalence of Cryptosporidium oocysts in fish species in Azerbaijan. HIGHLIGHTS Different fish species have been investigated from two territories of the Kura River.; Cryptosporidium and Eimeria spp. oocysts are present in fish species in Azerbaijan.; This is the first report of Cryptosporidium oocysts in fish in Azerbaijan.; Eimeria carpelli is a wide-speared species of fish in Azerbaijan.;

Background: Chicken coccidiosis is a protozoan disease that leads to considerable economic losses in the poultry industry. Live oocyst vaccination is currently the most effective measure for the prevention of coccidiosis. However, it provides limited protection with several drawbacks, such as poor immunological protection and potential reversion to virulence. Therefore, the development of effective and safe vaccines against chicken coccidiosis is still urgently needed. Methods: In this study, a novel oral vaccine against Eimeria tenella was developed by constructing a recombinant Lactobacillus plantarum (NC8) strain expressing the E. tenella RON2 protein. We administered recombinant L. plantarum orally at 3, 4 and 5 days of age and again at 17, 18 and 19 days of age. Meanwhile, each chick in the commercial vaccine group was immunized with 3 × 102 live oocysts of coccidia. A total of 5 × 104 sporulated oocysts of E. tenella were inoculated in each chicken at 30 days. Then, the immunoprotection effect was evaluated after E. tenella infection. Results: The results showed that the proportion of CD4+ and CD8+ T cells, the proliferative ability of spleen lymphocytes, inflammatory cytokine levels and specific antibody titers of chicks immunized with recombinant L. plantarum were significantly increased (P < 0.05). The relative body weight gains were increased and the number of oocysts per gram (OPG) was decreased after E. tenella challenge. Moreover, the lesion scores and histopathological cecum sections showed that recombinant L. plantarum can significantly relieve pathological damage in the cecum. The ACI was 170.89 in the recombinant L. plantarum group, which was higher than the 150.14 in the commercial vaccine group. Conclusions: These above results indicate that L. plantarum expressing RON2 improved humoral and cellular immunity and enhanced immunoprotection against E. tenella. The protective efficacy was superior to that of vaccination with the commercial live oocyst vaccine. This study suggests that recombinant L. plantarum expressing the RON2 protein provides a promising strategy for vaccine development against coccidiosis. [ABSTRACT FROM AUTHOR]

Background: Sarcocystis includes a global group of apicomplexan parasites with two-host life cycle frequently circulating in wildlife and domestic hosts, including humans. Two of the most important wild terrestrial carnivores acting as definitive hosts are the red fox and raccoon dog, due to their wide distribution in Europe and usage of wild and farmed animals as prey. This study was conducted to determine the prevalence of Sarcocystis in hunted red foxes and raccoon dogs from nine regions of the Czech Republic and to identify isolated sporocysts by molecular techniques. Methods: Approximately 5 g of the contents of large intestine from 200 animals (197 red foxes and three raccoon dogs) were examined by flotation centrifugation coprological method. Only samples of 50 red foxes and one raccoon dog positive to Sarcocystis spp. were used for the nested PCR (nPCR) method to amplify a fragment or partial sequence on the cox1 gene. Ten species-specific primer pairs for detection of Sarcocystis spp. using farm animals as intermediate hosts were utilized. Results: In total, 38.1% of the red foxes and 66.7% of the raccoon dogs were positive to Sarcocystis by light microscopy. The molecular characterization resulted in the identification of five species in the red fox: S. arieticanis, S. capracanis, S. cruzi, S. miescheriana, and S. tenella, while the PCR was negative for the sole raccoon dog. The highest intraspecific variation was found for S. miescheriana, while S. tenella was the most prevalent. Co-infections occurred in the large intestine of the red fox. No zoonotic species were found in our samples. Conclusion: This is the first study where the potential role of the red fox and raccoon dogs as spreaders of Sarcocystis to farm animals in the Czech Republic is shown. The use of species-specific primers provides a fast and easy method for screening multiple samples for a particular Sarcocystis species. [ABSTRACT FROM AUTHOR]

Background: Malaria transmission-blocking vaccines (TBVs) aim to inhibit malaria parasite development in mosquitoes and prevent further transmission to the human host. The putative-secreted ookinete protein 25 (PSOP25), highly conserved in Plasmodium spp., is a promising TBV target. Here, we investigated PvPSOP25 from P. vivax as a TBV candidate using transgenic murine parasite P. berghei and clinical P. vivax isolates. Methods and findings: A transgenic P. berghei line expressing PvPSOP25 (TrPvPSOP25Pb) was generated. Full-length PvPSOP25 was expressed in the yeast Pichia pastoris and used to immunize mice to obtain anti-rPvPSOP25 sera. The transmission-blocking activity of the anti-rPvPSOP25 sera was evaluated through in vitro assays and mosquito-feeding experiments. The antisera generated by immunization with rPvPSOP25 specifically recognized the native PvPSOP25 antigen expressed in TrPvPSOP25Pb ookinetes. In vitro assays showed that the immune sera significantly inhibited exflagellation and ookinete formation of the TrPvPSOP25Pb parasite. Mosquitoes feeding on mice infected with the transgenic parasite and passively transferred with the anti-rPvPSOP25 sera showed a 70.7% reduction in oocyst density compared to the control group. In a direct membrane feeding assay conducted with five clinical P. vivax isolates, the mouse anti-rPvPSOP25 antibodies significantly reduced the oocyst density while showing a negligible influence on mosquito infection prevalence. Conclusions: This study supported the feasibility of transgenic murine malaria parasites expressing P. vivax antigens as a useful tool for evaluating P. vivax TBV candidates. Meanwhile, the moderate transmission-reducing activity of the generated anti-rPvPSOP25 sera necessitates further research to optimize its efficacy. Author summary: The persistence of Plasmodium vivax poses a significant public health concern in certain regions, particularly Southeast Asia. The distinctive biology of Plasmodium vivax presents challenges for its control and eradication efforts. The development of a transmission-blocking vaccine is considered an essential strategy for malaria elimination, while the identification of antigen candidates plays a critical role in vaccine development. In this study, we aimed to evaluate the potential of PvPSOP25 as a transmission-blocking vaccine candidate using the transgenic murine parasite P. berghei. The transmission-reducing activity of anti-rPvPSOP25 sera was evaluated by both in vitro and mosquito-feeding experiments. Additionally, a direct membrane feeding assay using clinical P. vivax isolates from Thailand further validated the moderate transmission-reducing activity exhibited by the anti-rPvPSOP25 antisera. [ABSTRACT FROM AUTHOR]

Background: This study investigated effects of different methionine (Met) supplementation levels in a reduced protein diet on growth performance, intestinal health, and different physiological parameters in broilers under Eimeria challenge. A total of 600 fourteen-day-old Cobb500 male broilers were challenged with E. maxima, E. tenella, and E. acervulina, and randomly allocated in a 2 × 5 factorial arrangement. Birds received normal protein diets (20% crude protein, NCP) or reduced protein diets (17% crude protein, LCP), containing 2.8, 4.4, 6.0, 7.6, and 9.2 g/kg of Met. Results: On 6 and 9 days post inoculation (DPI), increasing Met level linearly improved the growth performance (P < 0.05). Total oocyst shedding linearly increased as Met level increased (P < 0.05). Duodenal villus height (VH):crypt depth (CD) in the LCP groups were higher on 6 DPI (P < 0.01) while lower on 9 DPI (P < 0.05) compared to the NCP groups. Jejunal CD and duodenal VH:CD changed quadratically as Met level increased (P < 0.05). On 6 DPI, liver glutathione (GSH) and glutathione disulfide (GSSG) linearly increased as Met level increased (P < 0.05). On 9 DPI, GSSG quadratically increased, whereas GSH:GSSG quadratically decreased as Met levels increased (P < 0.05). The expression of amino acid transporters linearly decreased as Met level increased (P < 0.05). The expression of zonula occludens 2 and claudin-1 linearly increased on 6 DPI whereas decreased on 9 DPI as Met level increased (P < 0.05). The expressions of cytokines were lower in the LCP groups than the NCP groups (P < 0.05). Interaction effects were found for the expression of IL-10 and TNFα on 6 DPI (P < 0.05), where it only changed quadratically in the NCP group as Met level increased. The expression of Met and folate metabolism genes were lower in the LCP groups than the NCP groups on 9 DPI (P < 0.05). The expression of these genes linearly or quadratically decreased as Met level increased (P < 0.05). Conclusion: These results revealed the regulatory roles of Met in different physiological parameters including oxidative status, intestinal health, and nutrient metabolism in birds fed reduced protein diet and challenged with Eimeria. [ABSTRACT FROM AUTHOR]

Introduction: Eimeria spp. are intracellular protozoan parasites of the phylum Apicomplexa causing economic losses to various wild and domestic animals. An eimerian species infecting Columba livia domestica was identified in this study. Methods: A total of 15 faecal samples were examined by floatation technique, a prevalence rate of 60% was reported. Eimerian oocysts were sporulated in 2.5% potassium dichromate solution then identified using morphological and molecular (DNA amplification of the 18S rRNA and ITS-1 genes) diagnostic techniques. Results: Sporulated oocysts were identified as Eimeria labbeana-like, after morphometry with typical bi-layered wall with spherical to subspherical oocysts morphology. A polar granule is present, but no micropyle or oocyst residuum. Sporocysts are elongated ovoidal with stieda body. Sporocyst residuum with many granules and sporozoites with refractile bodies and nucleus. Both 18S rRNA and ITS-1 sequences have been deposited in GenBank database. DNA sequences from the partial 18S rRNA generated from the oocysts were found to be related to eimerian and isosporan parasites found in domestic pigeons. For the first time, ITS-1 sequences for E. labbeana-like were provided. Conclusion: The necessity of using molecular techniques to describe pigeon intestinal coccidian parasites in conjunction with traditional morphology-based tools was emphasized in this work in order to understand the biology of such parasites. [ABSTRACT FROM AUTHOR]

Avian coccidiosis, a parasitic disease prevalent in poultry, is caused by Eimeria species and leads to significant economic losses. The use of attenuated live oocyst vaccines has been adopted as an alternative to the use of anticoccidial drugs. However, the accurate detection and differentiation of vaccine strains from virulent ones remain challenging. Therefore, this study presents a novel TaqMan polymerase chain reaction (PCR) detection method that offers enhanced sensitivity, specificity, and reproducibility compared with traditional PCR techniques. Through whole-genome resequencing and bioinformatics analysis, we identified a molecular marker gene, Em_marker6, with a unique 21-base pair deletion specific to the Eimeria maxima attenuated vaccine strain. Optimized primers and probes targeting this marker enabled rapid quantification cycle value achievement and high fluorescence intensity. The standard curve's slope of -3.540 and correlation coefficient of 0.9971 confirmed precise quantification capabilities. The TaqMan PCR method detected as few as 30 plasmid DNA copies and 50 oocysts per reaction, outperforming traditional PCR techniques by an order of magnitude. No cross-reactivity was observed with other E. maxima wide-type strains or common intestinal pathogens, ensuring the exclusive detection of the E. maxima EMPY vaccine strain. Weekly testing over 3 weeks demonstrated minimal variability, indicating robust consistency in the method's application. Testing on 61 clinical samples revealed a 57.38% positivity rate for E. maxima species and 13.11% for the vaccine strain. The Em_marker6 gene exhibited genetic stability across multiple generations, confirming the detection method's robust stability for the attenuated E. maxima vaccine strain. This study significantly advances the field of avian coccidiosis research and control by providing a valuable tool for monitoring vaccine purity and preventing inadvertent infections in vaccinated flocks, aligning with global efforts to curb antibiotic use in animal feed. [ABSTRACT FROM AUTHOR]

Rabbit coccidiosis is caused by infection with one or, more commonly, several Eimeria species that parasitize the hepatobiliary ducts or intestinal epithelium of rabbits. Currently, there are eleven internationally recognized species of rabbit coccidia, with the complete mitochondrial (mt) genomes of six species commonly infecting rabbits having been sequenced and annotated. Eimeria kongi was initially discovered in 2011 and prompted a preliminary study on this species. Through traditional morphological analysis, E. kongi was identified as a novel species of rabbit coccidia. To further validate this classification, we sequenced and annotated its mitochondrial genome. The complete mt genome of E. kongi spans 6258 bp and comprises three cytochrome genes (cytb, cox1, cox3), fourteen gene fragments for the large subunit (LSU) rRNA, and nine gene fragments for the small subunit (SSU) rRNA, lacking transfer RNA (tRNA) genes. Moreover, phylogenetic analysis of the mitochondrial genome sequence of E. kongi revealed its clustering with six other species of rabbit coccidia into a monophyletic group. Additionally, E. irresidua and E. flavescens were grouped within the lineage lacking oocyst residuum, consistent with their morphological characteristics. Consistent with multiple molecular phylogenies, in this investigation, E. kongi was further confirmed as a new species of rabbit coccidia. Our research findings are of great significance for the classification of coccidia and for coccidiosis prevention and control in rabbits. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the effect of Calotropis procera (Asclepiadaceae) extract on Eimeria piriformis (E. piriformis) oocysts and sporozoites (coccidiosis) isolated from commercial rabbits. Methods: Calotropis procera extract was obtained from dried, pulverized leaves, and macerated in 80 % methanol for 30 h. The extract was analyzed using infrared spectroscopy (IR). Twelve-well plates of 3 mL containing 1 x 104 non-sporulated oocyst were tested in 6 groups: Negative (untreated) control received 2.5 % potassium dichromate solution. Other groups received four concentrations of C. procera leaf extract (12.5, 25, 50, and 100 mg/mL) for oocysts vitality test. Toltrazuril (25 µl/mL) was used as positive control. Moreover, 250, 500, 750, and 1000 µg/mL of C. procera extract were tested for sporozoites vitality test. The mixture was examined daily for 4 days for oocysts vitality and after 12 and 24 h for sporozoite vitality. Results: Findings of IR showed that extracts contained 5 biologically active chemical components, indicating the presence of alkaloids, flavonoids, saponins, and phenols. The extract showed a significant inhibitory effect on E. piriformis oocysts at 100 mg/mL with suppression rates of approximately 89 % after 96 h, nearly similar to toltrazuril (p ≤ 0.01). Furthermore, C. procera extract showed inhibition at the highest repression (p ≤ 0.01) of 89 % of E. piriformis sporozoites viability at 1000 µg/mL after 24 h. The inhibitory rate increased proportionately with an extended incubation duration and high concentration. Conclusion: C. procera extract is as effective as the positive control in treating coccidiosis in rabbits. However, further studies are necessary to determine the active constituents of C. procera and their mode(s) of action. [ABSTRACT FROM AUTHOR]

Simple Summary: This study investigates the prevalence of gastrointestinal parasites in small ruminant farms (159 sheep and 39 goats) in southern Spain, a key area for the country's livestock production. This research reveals that Eimeria spp. Is the most common parasitic infection, followed by Strongyles. Other parasites, like Moniezia spp., Trichuris spp., and D. dendriticum, are less prevalent but more common in sheep. This study also includes a survey on farmers' management practices, highlighting that regular monitoring through coprological analyses is not common; veterinarians are seldom involved in deworming plans, and anthelmintic treatment is typically applied twice a year in sheep and once in goats. This report suggests that implementing certain management measures could potentially mitigate parasite infections. This constitutes the first report on the epidemiological status of gastrointestinal parasites in small ruminants in southern Spain. The primary population of small ruminants in Spain is concentrated in the southern region, a critical area for the country's livestock production. Indirect economic losses can occur when this livestock is affected by gastrointestinal parasites. This study aimed to determine the prevalence of these parasites in small ruminant herds (159 sheep and 39 goats) through coprological analyses and conducted a survey on farmers' management practices related to gastrointestinal parasite control. The survey results revealed some important aspects: monitoring through coprological analyses is not a common practice; veterinarians are not typically involved in deworming plans; anthelmintic treatment in adults is often applied twice a year in sheep and once a year in goats; and finally, drug rotation was higher in sheep farms. Coprological analyses showed Eimeria spp. as the most common parasitic infection, followed by Strongyles infection. Other parasites like Moniezia spp., Trichuris spp., and D. dendriticum were less important, although their prevalence was higher in sheep than goats. This constitutes the first report on the epidemiological status of gastrointestinal parasites in small ruminants in southern Spain. Based on the survey findings, the introduction of certain management measures on farms could potentially mitigate parasite infections. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Quails are an important alternative to chicken production for protein sources, offering many advantages over other poultry species. However, raising quail faces certain challenges, such as a shortage of specified hatcheries and the lack of local markets for quail eggs and meat, particularly in Egypt. In addition, there is less interest in quail's medication and vaccine production. A significant disease affecting the health and productivity of quails is coccidiosis, which is associated with poor feed conversion ratio, lower growth performance, heightened mortality, and high cost of vaccination and treatment. Attention to quail coccidiosis and its clinical forms, diagnosis, morphological characterization, control, and prevention is very critical for improving quail meat and egg production. This review compiles scientific data on quail coccidiosis, with a focus on literature from Egypt, for classification, data analysis, and processing. [ABSTRACT FROM AUTHOR]

The animals held captive in zoos often face health and well-being issues. Parasitic infections can lead to health problems in wildlife animals by affecting their gastrointestinal tract. Therefore, the present study aimed to identify and evaluate the population of the various Gastrointestinal (GIT) parasites of wild animals enclosed in different zoological gardens in Pakistan. The fresh fecal samples (n = 960) of 20 captive wildlife animals were collected from Marghzar Zoo, Islamabad (n = 340), Ayub National Park, Rawalpindi (n = 221), Lohi Bher Wildlife Park, Rawalpindi (n = 296), and Bansra Galli Wildlife Park, Rawalpindi (n = 103). The samples were obtained from wildlife mammals, including urial (n = 95), blue bull (n = 106), chinkara gazelle (n = 77), zebra (n = 77), hog deer (n = 75), spotted deer (n = 43), blackbuck (n = 58), barking deer (n = 52), red deer (n = 104), yak (n = 44), grey goral (n = 40), lion (n = 37), mouflon sheep (n = 46), red fox (n = 12), bear (n = 37), grey wolf (n = 12), jackal (n = 12), vervet monkey (n = 12), rhesus monkey (n = 12), and langoor (n = 12). Various methods, such as direct smear examination, standard sedimentation, and floatation techniques were applied to detect and identify the endoparasites in the fecal sample. The detailed routine parasitological analysis identified approximately 52 endoparasites in the fecal samples, including Haemonchus contortus, Eimeria bovis, Ostertagia curcumcincta, Strongyloides papillosus, Strogylus equinus, Oxyuris equi, Chabertia ovina, Protostrongylus, and Trichostrongylus vitrines. The obtained results indicated that Lohi Bher Wildlife Park (46.35%) had a higher prevalence of GIT parasites, compared to Marghzar Zoo (33.23%), Bansra Galli Wildlife Park (33.02%), and Ayub National Park (19.45%). The study reports mild to moderate parasitic infection in captive wild animals and that could affect the survivability of the animals in captivity. The findings of the study can be used to formulate a proper health protocol and sanitation management in captive wild animals to control parasitic infections. [ABSTRACT FROM AUTHOR]

The camel has played a role in human civilization since its inception and holds significant importance in the customs and agricultural practices of various nations. This study examined the prevalence of internal parasitic infestations in camels within the Al-Diwaniyah and Al-Najaf provinces of Iraq from December 2021 to September 2022. A total of 200 fecal samples were randomly collected from farm camels, revealing that these animals were affected by one or more types of intestinal parasites. Nematodes exhibited the highest prevalence at 56 %, followed by Protozoa at 28.5 %, Cestodes at 14.5 %, and Trematodes at 1 %. Among these parasites, Trichostrongylus spp. had the highest percentage at 33 %, followed by Moneizia benedeni (12.5 %), Fasciola hepatica (10.5 %), Strongyloides spp. (8 %), Giardia spp. (7 %), Nematodirus spp. (6 %), and Eimeria spp. (6 %). Furthermore, mixed-species or single-species infections in camels were observed, including Anoplcephala perfoliata (4 %), Haemonchus spp. (3.5 %), Dictyocaulus spp. (3 %), Trichuris trichura (2.5 %), Entamoeba spp. (2 %), and Balantidium coli (1 %). Additionally, nested PCR was employed to identify Trichostrongylus spp., with 45.4 % of camels testing positive for this particular parasite. [ABSTRACT FROM AUTHOR]

Background: Coccidiosis outbreaks in susceptible laying hens can significantly decrease egg production and cause substantial economic loss to the egg industry. The supplementation of poultry diets with chemotherapeutic agents is limited due to antimicrobial resistance and residue in poultry meat or processed products. Therefore, alternative strategies to control coccidiosis are needed, and Artemisia annua (AA) might have the potential to be a phytogenic feed additive, an alternative to anticoccidial agents. This study aimed to investigate the effect of the dietary supplementation of powdered AA on the performance and gut health of laying hens infected with coccidiosis by Eimeria spp. Methods: A total of 225 Hy-Line W-36 laying hens at 23 weeks of age were allocated into 5 treatment groups: 1) control (NC), 2) pair-fed (PF) control, 3) challenged control (CC), 4) CC with dietary inclusion of 0.5% AA (0.5AA), and v) CC with dietary inclusion of 1% AA (1AA). The hens in the CC, 0.5AA, and 1AA groups were orally inoculated with sporulated oocysts of Eimeria maxima (12,500), Eimeria tenella (12,500), and Eimeria acervulina (62,500) at week 25. The PF hens received the same amount of feed consumed by the CC hens from 0--14 days post-inoculation (dpi) of Eimeria spp. The performance of the laying hens, including body weight (BW), hen--day egg production (HDEP), feed intake (FI), and feed conversion ratio (FCR), was measured weekly. Additionally, markers of intestinal health, including gut permeability, lesion score, intestinal morphometry, and immune responses, were evaluated at 6, 14, and 21 dpi. Results: At 6 and 14 dpi, laying hens challenged with Eimeria spp. had a lower BW than PF and NC hens (p < 0.0001). Supplementation of 1% AA improved the HDEP by 8.1% compared to CC hens; however, it was still 15.4% lower than that of PF hens (p < 0.0001). The inclusion of 1% AA did not have any beneficial effect on FI; however, the FCR was improved by 0.61 (2.46) than that of CC hens (3.07; p < 0.0001). The inclusion of 1% AA reduced the severity of the intestinal lesions and increased the recovery of intestinal villi (p < 0.05). Additionally, gut permeability was significantly different between the challenged and non-challenged hens; however, among the challenged hens, the inclusion of AA reduced the gut permeability by 29% compared to CC hens (p < 0.0001). Furthermore, the inclusion of 0.5% AA reduced the inflammatory responses in the infected hens. Conclusion: Dietary inclusion of AA partially restored the performance and gut health of the laying hens and modulated their inflammatory immune response following Eimeria infection; however, further studies are needed to better understand the mode of action and effective dosages to improve the gut health without negative impacts on the performance. [ABSTRACT FROM AUTHOR]

Coccidiosis, an intestinal disease caused by Eimeria parasites, is responsible for major losses in the poultry industry by impacting chicken health. The gut microbiota is associated with health factors, such as nutrient exchange and immune system modulation, requiring understanding on the effects of Eimeria infection on the gut microbiota. This study aimed to determine the effects of Eimeria acervulina infection on the luminal and mucosal microbiota of the cecum (CeL and CeM) and ileum (IlL and IlM) at multiple time points (days 3, 5, 7, 10, and 14) post-infection. E. acervulina infection decreased evenness in CeL microbiota at day 10, increased richness in CeM microbiota at day 3 before decreasing richness at day 14, and decreased richness in IlL microbiota from day 3 to 10. CeL, CeM, and IlL microbiota differed between infected and control birds based on beta diversity at varying time points. Infection reduced relative abundance of bacterial taxa and some predicted metabolic pathways known for short-chain fatty acid production in CeL, CeM, and IlL microbiota, but further understanding of metabolic function is required. Despite E. acervulina primarily targeting the duodenum, our findings demonstrate the infection can impact bacterial diversity and abundance in the cecal and ileal microbiota. [ABSTRACT FROM AUTHOR]

The objectives of this study were to identify the species of psittaciform birds kept in captivity as companion and ornamental animals in Yucatán, Mexico, as well as to estimate the prevalence and intensity of the intestinal parasites. 16 species of psittaciform belonging to 11 different genera were studied, of which three species are locally distributed (Amazona albifrons, Amazona xantholora and Eupsittula nana astec), two are introduced (Amazona autumnalis and Amazona oratrix) and 11 are exotic (Agapornis sp., Amazona auropalliata, Amazona farinosa, Aratinga solstitialis, Barnardius zonaris, Cyanoramphus novaezelandiae, Melopsittacus undulatus, Noephema bouerkii, Nymphicus hollandicus, Psephotus sp. and Psittacula krameri) for the state of Yucatán, Mexico. A total of 84 stool samples were obtained and processed using the centrifugal flotation and McMaster techniques. A prevalence of 14.28 % of birds with intestinal parasites was estimated, reporting the presence of the nematodes Ascaridia spp. (33.3 %), Capillaria spp. (16.6 %) and Heterakis spp. (16.6 %), cestodes Hymenolepis spp. (16.6 %) and Raillietina spp. (8.3 %) and the protozoan Eimeria spp. (16.6 %). The intensity of eggs and oocysts per gram of feces (h/gh and ooq/gh) was from <50 to 600, with high intensities only observed in exotic species such as B. zonaris and C. novaezelandiae. It is concluded that six genera of intestinal parasites were present in psittaciforms from Yucatán, Mexico with nematodes being the ones that appeared most frequently. The highest intensities occurred in exotic bird species, being important in the implementation of deworming and quarantine protocols to prevent parasitic infections in birds. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Coccidiosis is one of the most frequently reported diseases in chickens, causing a significant economic impact on the poultry industry. However, there have been no previous studies evaluating the prevalence of this disease in broiler farms in Guangdong province. Therefore, this study aims to conduct an epidemiological investigation into the occurrence of Eimeria species and associated risk factors in intensive management conditions across four regions in Guangdong province, China. A total of 394 fecal samples were collected from 89 broiler farms in Guangdong province. The prevalence of Eimeria species infection was determined using PCR, and the occurrence of Clostridium perfringens type A was assessed using quantitative real-time PCR. Results: The results showed an overall prevalence of 98.88% (88/89) at the farm level and 87.06% (343/394) at the flock level. All seven Eimeria species were identified, with E. acervulina (72.53%; 64/89), E. tenella (68.54%; 61/89), and E. mitis (66.29%; 59/89) at the farm level, and E. acervulina (36.55%; 144/394), E. mitis (35.28%; 139/394), and E. tenella (34.01%; 134/394) at the flock level. The predominant species combination observed was a co-infection of all seven Eimeria species (6.74%; 6/89), followed by a combination of E. acervulina, E. tenella, E. mitis, E. necatrix, E. brunetti, and E. maxima (5.62%, 5/89). A combination of E. acervulina, E. tenella, E. mitis, E. necatrix, E. brunetti, and E. praecox (4.49%; 4/89) was also observed at the farm level. Furthermore, the study identified several potential risk factors associated with the prevalence of Eimeria species, including farm location, chicken age, drinking water source, control strategy, and the presence of C. perfringens type A were identified as potential risk factors associated with prevalence of Eimeria species. Univariate and multivariate analyses revealed a significant association between E. necatrix infection and both grower chickens (OR = 10.86; 95% CI: 1.92–61.36; p < 0.05) and adult chickens (OR = 24.97; 95% CI: 4.29–145.15; p < 0.001) compared to starter chickens at the farm level. Additionally, farms that used groundwater (OR = 0.27; 95% CI: 0.08–0.94; p < 0.05) were less likely to have E. maxima compared to those that used running water. At the flock level, the prevalence of E. tenella was significantly higher in the Pearl River Delta (OR = 2.48; 95% CI: 1.0–6.15; p = 0.05) compared to eastern Guangdong. Interestingly, flocks with indigenous birds were less likely to have E. brunetti (OR = 0.48; 95% CI: 0.26–0.89; p < 0.05) compared to flocks with indigenous crossbred birds. Furthermore, flocks that used anticoccidial drugs (OR = 0.09; 95% CI: 0.03–0.31; p < 0.001) or a combination of vaccines and anticoccidial drugs (OR = 0.06; 95% CI: 0.01–0.25; p < 0.001) were less likely to be positive for E. tenella compared to flocks that only used vaccines. Finally, flocks with C. perfringens type A infection were significantly more likely to have E. necatrix (OR = 3.26; 95% CI: 1.96–5.43; p < 0.001), E. tenella (OR = 2.14; 95% CI: 1.36–3.36; p < 0.001), E. brunetti (OR = 2.48; 95% CI: 1.45–4.23; p < 0.001), and E. acervulina (OR = 2.62; 95% CI: 1.69–4.06; p < 0.001) compared to flocks without C. perfringens type A. Conclusions: This study conducted an investigation on the prevalence, distribution, and risk factors associated with Eimeria species infection in broiler chickens in Guangdong. The farm-level prevalence of Eimeria species was higher than the previous prevalence figures for other areas and countries. E. brunetti was identified at higher prevalence in Guangdong than previously survived prevalence in different regions in China. Farm location, chicken age, drinking water source, control strategy, and the presence of C. perfringens type A were considered as potential risk factors associated with prevalence of Eimeria species. It is imperative to underscore the necessity for further surveys to delve deeper into the occurrence of Eimeria species under intensive management conditions for different flock purposes. [ABSTRACT FROM AUTHOR]

Willow (Salix spp.) trees, found worldwide, contain secondary metabolites that are valuable as dietary supplements for animal feed and as antiparasitic compounds. We quantified secondary metabolites (phenolics, flavonoids, and salicylic acid) in ethanolic extracts from leaves and branches of three Salix acmophylla Boiss. genotypes and investigated their potential to inhibit Eimeria sp. sporulation, a major concern in ruminants. The total phenolic content of willow leaves and branches was similar in two of three different genotypes. The total flavonoid content of the branches was significantly higher than that of leaves of the same genotype; however, the salicylic acid content was significantly higher in leaves than in branches. Importantly, all extracts exhibited significant inhibition of Eimeria sporulation, where over 70% inhibition was obtained at concentrations as low as 750 mgL−1. The sporulation inhibition by branch or leaf extracts exceeded 80% for leaves and 90% for branches at concentrations above 1250 mgL−1. The study highlights the potential of using Salix extracts as bioactive compounds for biological control of coccidiosis in ruminants. We conclude that all parts and all investigated genotypes of S. acmophylla can provide secondary metabolites that act as a coccidiostat to treat Eimeria in goats. [ABSTRACT FROM AUTHOR]

Within-host interactions among coinfecting parasites can have major consequences for individual infection risk and disease severity. However, the impact of these within-host interactions on between-host parasite transmission, and the spatial scales over which they occur, remain unknown. We developed and apply a novel spatially explicit analysis to parasite infection data from a wild wood mouse (Apodemus sylvaticus) population. We previously demonstrated a strong within-host negative interaction between two wood mouse gastrointestinal parasites, the nematode Heligmosomoides polygyrus and the coccidian Eimeria hungaryensis, using drug-treatment experiments. Here, we show this negative within-host interaction can significantly alter the between-host transmission dynamics of E. hungaryensis, but only within spatially restricted neighbourhoods around each host. However, for the closely related species E. apionodes, which experiments show does not interact strongly with H. polygyrus, we did not find any effect on transmission over any spatial scale. Our results demonstrate that the effects of within-host coinfection interactions can ripple out beyond each host to alter the transmission dynamics of the parasites, but only over local scales that likely reflect the spatial dimension of transmission. Hence there may be knock-on consequences of drug treatments impacting the transmission of non-target parasites, altering infection risks even for non-treated individuals in the wider neighbourhood. [ABSTRACT FROM AUTHOR]

Chicken coccidiosis caused by Eimeria spp. can occur on almost all poultry farms, causing huge economic losses to the industry. Genetically manipulated Eimeria parasites as a vaccine vector to deliver viral antigens have been reported. In our preliminary study, transgenic E. acervulina expressing a VP2 gene (Ea-VP2) of the infectious bursal disease virus (IBDV) demonstrated partial protection against IBDV infection. To enhance immune responses, we aimed to increase the VP2 gene copy number in transgenic E. acervulina. In this study, we used a novel plasmid vector carrying a VP2 gene fused with three flag tags and a red fluorescent reporter gene (mCherry). The vector was introduced into Ea-VP2 sporozoites through nucleofection, leading to the generation of Ea-2VP2. Subsequent analysis revealed a notable escalation in the fluorescent rate, increasing from 0.11 to 95.1% following four consecutive passages facilitated by fluorescent-activated cell sorting. Verification via PCR, Western blot, and immunofluorescence confirmed the successful construction of the Ea-2VP2 population. Despite lower fecundity compared to wild-type E. acervulina, Ea-2VP2 maintained immunogenicity. Our research effectively created a transgenic E. acervulina strain transfected sequentially with two copies of the VP2 gene from IBDV. This modification resulted in an increased humoral immune response after primary immunization in chickens. Additionally, it demonstrated a degree of protection within the bursa against IBDV infection. Future studies will focus on further enhancing immune response levels. [ABSTRACT FROM AUTHOR]

This study aims to determine the prevalence of Cryptosporidium and Eimeria spp. oocysts in fish specimens in the river Kura. It was conducted during the 2021-2022 at two sites: Mingachevir reservoir in central Azerbaijan and in Neftchala district where the river finally enters the Caspian Sea through a delta of the Kura River estuary. The diagnosis of oocysts was performed microscopically. Fine smears from the intestine epithelial layers stained by Ziehl-Neelsen for Cryptosporidium oocysts. To identify Eimeria oocysts, each fish’s faecal material and intestinal scrapings were examined directly under a light microscope in wet samples on glass slides with a coverslip. Results revealed a prevalence of Cryptosporidium and Eimeria species infections in fish hosts from both territories Rutilus caspicus, Alburnus filippi, Abramis brama orientalis and Carassius gibelio. Of 170 investigated fish specimens, 8.8% (15/170) were infected with Cryptosporidium species oocysts. Eimeria species oocysts were identified in 20.6% (35/170). The presence of Cryptosporidium and Eimeria infections in fish specimens are natural infections. However, their presence in fish species may be attributed to the age of the fish species and water pollution. This is the first report regarding the prevalence of Cryptosporidium oocysts in fish species in Azerbaijan. [ABSTRACT FROM AUTHOR]

Eimeria are the absolute intestinal parasites belonging to phylum Apicomplexa, causing the intestinal disease known as coccidiosis. It is a lethal disease that affects a large variety of animals and birds including cattle, camel, goats, sheep, poultry, etc. causing severe devastation and mortalities. Controlling poultry coccidiosis by the means of preventive and prophylactic use of chemical drugs is currently being practiced on a commercial scale. Regardless of a history of successful use of these drugs, resistance and public health concerns demand their alternatives. Several alternatives including vaccination, organic acids and botanicals, are being suggested. Several preparations of botanicals have been experimented and found useful, but exact compound identification is a question of interest for scientists. Phenolic compounds are among the most abundant phytochemicals found in diverse structural forms comprising mainly phenolic acids, tannins, flavonoids, and others. Research states that they have the potential to control coccidiosis because of their direct and indirect anticoccidial activities. Phenolics have been reported to control coccidiosis by targeting Eimeria species directly and reducing the damages by antioxidant, immunomodulatory, and anti-inflammatory properties. Phenolics kill Eimeria by engulfing schizonts, rupturing the cell membrane of sporozoites, and inhibiting growth by stopping ATP synthesis mechanisms and other pathways depending on their chemical nature and the subgroup of phenolic compounds. Because of the high potential of being oxidized; they capture the reactive species produced because of cell rupture during rapid schizogony and immune invasion. Phenolics give negative feedback to several inflammatory mediators and reduce the inflammatory destruction of the gut, hence minimizing the loss. Regardless of these activities, phenolics have some drawbacks i.e., they reduce nutrient uptake, disturb carbohydrate disturbance, have cytotoxic effects, and undefined long-term response. Extensive research is needed to investigate the proper anticoccidial compound and reduce the side effects of phenolics to manage coccidiosis in an accurate, rapid, and safe way. [ABSTRACT FROM AUTHOR]

Coccidiosis is an important disease in juvenile fish because of severe intestinal injury during infection. We first reported the mixed infection of intestinal coccidia and its association with health status and pathological findings in juvenile Asian seabass (Lates calcarifer) cultured in Thailand. Two groups of Asian seabass, 60‐day fish and 90‐day fish, were sampled to investigate prevalence and coccidian infection intensity using morphological characterization and PCR. Phylogenetic analysis of 18S rRNA gene amplified from the intestines revealed Eimeria sp. and Cryptosporidium sp. infection. The prevalence of Eimeria sp. and Cryptosporidium sp. in sampled fish was 100%. Clinical outcomes assessed, using health assessment index (HAI) scoring and semi‐quantitative grading of intestinal lesions and inflammation, demonstrated that all fish developed variety of pathology and clinical illness; however, infection intensity in 60‐day fish was significantly higher (p <.05) than 90‐day fish. The HAI score of 60‐day fish was poorer than 90‐day fish, which correlated to a high infection intensity (r =.397), analysed by Pearson correlation coefficient. Overproduction of intestinal oxidants contributing to mucosal injury was examined by nitrotyrosine expression. The high production of reactive nitrogen species indicated severe inflammatory response, and intestinal injuries occurred mainly in the 60‐day fish. [ABSTRACT FROM AUTHOR]

Background: Ethno-veterinary practices could be used as a sustainable developmental tool by integrating traditional phytotherapy and husbandry. Phytotherapeutics are available and used worldwide. However, evidence of their antiparasitic efficacy is currently very limited. Parasitic diseases have a considerable effect on pig production, causing economic losses due to high morbidity and mortality. In this respect, especially smallholders and organic producers face severe challenges. Parasites, as disease causing agents, often outcompete other pathogens in such extensive production systems. A total of 720 faecal samples were collected in two farms from three age categories, i.e. weaners, fatteners, and sows. Flotation (Willis and McMaster method), modified Ziehl–Neelsen stained faecal smear, centrifugal sedimentation, modified Blagg technique, and faecal cultures were used to identify parasites and quantify the parasitic load. Results: The examination confirmed the presence of infections with Eimeria spp., Cryptosporidium spp., Balantioides coli (syn. Balantidium coli), Ascaris suum, Oesophagostomum spp., Strongyloides ransomi, and Trichuris suis, distributed based on age category. A dose of 180 mg/kg bw/day of Allium sativum L. and 90 mg/kg bw/day of Artemisia absinthium L. powders, administered for 10 consecutive days, revealed a strong, taxonomy-based antiprotozoal and anthelmintic activity. Conclusions: The results highlighted the therapeutic potential of both A. sativum and A. absinthium against gastrointestinal parasites in pigs. Their therapeutic effectiveness may be attributed to the content in polyphenols, tocopherols, flavonoids, sterols, sesquiterpene lactones, and sulfoxide. Further research is required to establish the minimal effective dose of both plants against digestive parasites in pigs. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the prevalence of Eimeria species in domestic pigeons and study the gross as well as the microscopic pathology they cause in the intestinal tract of the birds in Makurdi, Benue State. A total of 310 healthy pigeons comprising 171(55.2%) males and 139(44.8%) females; 133(42.9%) were sampled during the dry season and 177(57.1%) were sampled during the wet season from local farmers in Makurdi, Benue State. Faeces from the rectum of each bird were analyzed using the simple flotation method for the detection of Eimeria oocyst. Intestinal gross lesions were noted and intestinal segments were obtained for Histopathological studies. The prevalence of Eimeria species in pigeons of Makurdi was 170/310(54.8%). The infection was significantly higher (p<0.05) in the male Pigeons as compared to the female (p>0.05). There was no significant difference (p>0.05) between the number of pigeons infected in the wet season as compared with the dry season. Gross lesions revealed hemorrhagic enteritis and from the histopathological sections of the intestine, developmental stages of Eimeria species were seen in the mucosa, submucosa, and muscularis mucosa respectively with necrosis of villi and inflammatory cell infiltration. It was concluded that Since Eimeria species are highly prevalent in domestic pigeons and cause severe intestinal lesions, constant Veterinary surveillance is required to prevent economic losses in the pigeon industry. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine the prevalence and to identify the risk factors of Eimeria spp. and Giardia spp. infection in rabbits from the local population of four localities in northern Algeria. Dung samples were collected from 16 farms, totalling 111 rabbits, and were analysed by the flotation method. Additional, data regarding the farms and management practices were obtained by means of a questionnaire used in the surveys and interviews. The results revealed that the prevalence of Eimeriaspp. contamination was 68.75% (11/16) for farms and 58.56% (65/111) for rabbits, respectively. The prevalence of Giardia spp. was respectively 56.25% (9/16) for farms and 11.7% (13/111) for rabbits. The analyses showed that the prevalence of Eimeria spp. was significantly higher in the farms that did not comply with hygiene and non-conventional feeding and watering. However, the prevalence of Giardia spp. was significant in rabbits kept in poor conditions of rearing. In conclusion, this study showed that the prevalence of these two parasites in rabbits from the local population is relevant and may have important implications for the rabbit industry and public health, especially in rural areas. [ABSTRACT FROM AUTHOR]

ABSTRACT: A 35-d study investigated the impact of dietary supplementation with Arginine (Arg) or branched-chain amino acids (BCAA) of broilers receiving low-protein diets whilst infected with mixed Eimeria species. All birds were given the same starter (d0–10) and finisher (d28–35) diets. The 4 grower diets used were a positive control (PC) with adequate protein (18.5%), a low protein diet (NC;16.5% CP), or the NC supplemented with Arg or BCAA. Supplemental AA was added at 50% above the recommended levels. The treatments were in a 4 × 2 factorial arrangement, with 4 diets, with or without Eimeria inoculation on d14. Birds and feed were weighed after inoculation in phases: prepatent (d14–17), acute (d18–21), recovery (d22–28), and compensatory (d29–35). Ileal digesta, jejunum, and breast tissue were collected on d21, 28, and 35. There was no diet × Eimeria inoculation on growth performance at any phase. Infected birds weighed less and consumed less feed (P < 0.05) in all phases. In the prepatent and acute phases, birds on the Arg diets had higher weight gain (P < 0.05) and lower FCR, similar to PC, when compared to NC and BCAA-fed ones. Infection reduced AA digestibility on d21 and 28 (Met and Cys). However, birds that received supplemental AA had higher digestibility (P < 0.05) of their respective supplemented AA on d 21 only. Infected birds had lower (P < 0.05) BO + AT and higher PEPT1 expression on d21. There was a diet × Eimeria interaction (P = 0.004) on gene expression at d28; 4EBP1 genes were significantly downwardly expressed (P < 0.05) in birds fed Arg diet, irrespective of infection. Infected birds exhibited an upward expression (P < 0.05) of Eef2 on d21 and d28 but experienced a downward expression on d35. Supplemental Arg and BCAA had variable effects on growth performance, apparent ileal AA digestibility, and genes of protein synthesis and degradation, but the effect of Arg on promoting weight gain, irrespective of the Eimeria challenge, was more consistent.

For years, there has been an increasing interest in natural alternatives to the conventional coccidiostats applied as feed additives, which have been used for decades to prevent coccidiosis in poultry and fattening rabbits. This study aimed to compare the possible anticoccidial effects of oregano oil to the established substance diclazuril in growing rabbits. The control group (CG) received a non-supplemented basal compound feed, to which either diclazuril (1 mg/kg; DG) or oregano oil (75 mg/kg; OG) was added. In each of the three trials, subgroups of 50 rabbits each were assigned to one of the three experimental groups (CG, DG and OG). Natural Eimeria infection was monitored weekly by fecal oocyst counts and Eimeria species identification following sporulation. Additionally, the performance parameters were determined at the middle and the end of the trials, and the deceased rabbits were subjected to necropsy. Neither oocyst excretion nor the performance parameters differed significantly between the three experimental groups. Eimeria media, Eimeria magna, Eimeria perforans and Eimeria exigua were identified as the occurring species. The highest animal losses (16.0%) occurred in the OG, while the losses were 12.7% in the DG and 12.0% in the CG. However, these differences were not statistically significant. Overall, neither diclazuril nor oregano oil was superior to the non-supplemented feed. This underlines the importance of diagnostics, as this study’s results indicate that in the absence of the highly pathogenic Eimeria species, economic rabbit rearing and fattening is achievable without the use of coccidiostats.

Abstract Eimeria species serve as promising eukaryotic vaccine vectors. And that the location of heterologous antigens in the subcellular components of genetically modified Eimeria may determine the magnitude and type of immune responses. Therefore, our study aimed to target a heterologous fluorescent protein to the cell surface or microneme, two locations where are more effective in inducing protective immunity, of Eimeria tenella and E. acervulina sporozoites. We used an enhanced yellow fluorescent protein (EYFP) as a tagging biomarker, fusing variously with some localization or whole sequences of compartmental proteins for targeting. After acquiring stable transgenic Eimeria populations, we observed EYFP expressing in expected locations with certain strategies. That is, EYFP successfully localized to the surface when it was fused between signal peptides and mature products of surface antigen 1 (SAG1). Furthermore, EYFP was efficiently targeted to the apical end, an optimal location for secretory organelle known as the microneme, when fused to the C terminus of microneme protein 2. Unexpectedly, EYFP exhibited dominantly in the apical end with only weak expression on the surface of the transgenic sporozoites when the parasites were transfected with plasmid with EYFP fused between signal peptides and mature products of E. tenella SAG 13. These strategies worked in both E. tenella and E. acervulina, laying a solid foundation for studying E. tenella and E. acervulina-based live vaccines that can be further tailored to the inclusion of cargo immunogens from other pathogens.

Irrigation water contaminated by human fecal material may elevate the risk of produce contamination with the enteric parasite Cyclospora cayetanensis. Oocysts of C. cayetanensis are resistant to commonly used disinfectants and a method of removing C. cayetanensis from irrigation water would mitigate this risk. We evaluated zero valent iron (ZVI) sand filtration as one such method. We sought to determine if sand filters containing ZVI outperformed those without ZVI. We first evaluated the abundant poultry parasites Eimeria maxima, E. tenella and E. acervulina as surrogates for C. cayetanensis. We determined if a miniaturized gravity fed ZVI-sand filter, scaled to evaluate scarce supplies of C. cayetanensis oocysts, provided useful information about the performance of larger filtration systems. Filters were inoculated with oocysts, rinsed, and the resulting filtrate examined microscopically for oocysts. We performed experiments to measure the effect of varying ZVI concentrations, repeated filter use, simulated agricultural water, and oocyst size and condition. We then compared the performance of mini filters to that of larger, gravity-fed pool filters and found that ZVI-sand filtration was far more effective at removing Eimeria spp. from water when compared to sand filtration, at both scales. Sand mini filters retained 13–54 % of E. acervulina oocysts, and pool filters retained 82 %, but when combined with 50 % (mini filter) or 35 % (pool filter) v/v ZVI, mini filters retained 89–99 % of oocysts and pool filters retained >99 %. The effectiveness of the mini filters increased with increasing ZVI concentration, and the addition of ZVI far outweighed the influence of any other measured variable. We then performed experiments including C. cayetanensis, which provided similar results to those utilizing Eimeria; 59 % of inoculated C. cayetanensis oocysts were retained in sand mini filters, and 97 % in mini filters composed of 35 % v/v ZVI. In sum, ZVI is highly effective in removing oocysts from water and Eimeria is a useful surrogate for C. cayetanensis to assess filtration. ZVI-sand filtration shows promise as a tool to mitigate the risk of C. cayetanensis contamination of irrigation water. Further studies should evaluate the performance of ZVI-sand in pressurized fast filtration systems under a range of field conditions.