Eimeria Species Research Articles

Morphological and molecular characterization of Isospora elliotae n. sp. (Apicomplexa: Eimeriidae) from the Australian magpie Gymnorhina tibicen (Latham, 1801) (Passeriformes: Artamidae) in Western Australia.

A new coccidian species, Isospora elliotae n. sp., from the Australian magpie Gymnorhina tibicen (Latham, 1801) in Western Australia, is described and characterized morphologically and molecularly. Microscopic analysis of a faecal sample identified subspheroidal oocysts (n = 20), 20-22 × 18-20 (20.7 × 18.7); length/width (L/W) ratio 1.05-1.14 (1.10). Wall bi-layered, 1.0-1.3 (1.2) thick, outer layer smooth, c. 2/3 of total thickness. Micropyle and oocyst residuum absent, but usually two polar granules are present. Sporocysts (n = 28) ovoidal, 12-13 × 9-11 (12.6 × 9.7); L/W ratio 1.22-1.35 (1.30). Stieda body present, flattened to half-moon-shaped, c. 0.5 deep × 2.0 wide; sub-Stieda indistinct or barely discernible, c. 1.0 deep × 2.5 wide; para-Stieda body absent; sporocyst residuum present, composed of granules dispersed among the sporozoites. Sporozoites vermiform, with anterior and posterior refractile bodies and nucleus. Segments of three gene loci (18S rRNA, 28S rRNA and COI) were sequenced and I. elliotae n. sp. exhibited 99.8% genetic similarity to Isospora sp. MAH-2013a (KF648870) followed by 99.7% genetic similarity to Isospora neochmiae (Yang, Brice & Ryan, 2016) (KT224380) at the 18S rRNA gene locus. It shared 97.0% genetic similarity with an unnamed Isospora sp. (AY283852) at the 28S rRNA gene locus and it also shared the highest genetic similarity of 99.8% with the unnamed Isospora sp. from an American crow (OL999120) at the COI gene locus. Based on morphological and molecular data, this isolate is a new species named as I. elliotae n. sp.

Brief report: Isospora gilvusi n. sp. (Apicomplexa: Eimeriidae) from the warbling vireo Vireo gilvus Vieillot (Passeriformes: Vireonidae) in Morelia, Mexico.

A new coccidian species, Isospora gilvusi n. sp. (Apicomplexa: Eimeriidae) collected from the warbling vireo Vireo gilvus, is reported from Morelia, Michoacán State, Mexico. Sporulated oöcysts of the new species are spherical to subspherical, 27-31 × 27-29 (30.1 × 28.4) μm, with a length/width (L/W) ratio of 1.1; one or two polar granules are present, but micropyle and oöcyst residuum are absent. Sporocyts are ovoid to drop-shaped, 16-17 × 11-12 (17.0 × 11.5) μm, with a L/W ratio of 1.7; Stieda and sub-Stieda bodies are both present, but para-Stieda body is absent; sporocyst residuum diffuse. At the histological study, endogenous stages were observed in the epithelial cells of the duodenum. This is the second species of Isospora recorded infecting a bird of the family Vireonidae in the New World.

Management and control of coccidiosis in poultry - A review.

Poultry coccidiosis is an intestinal infection caused by an intracellular parasitic protozoan of the genus Eimeria. Coccidia-induced gastrointestinal inflammation results in large economic losses, hence finding methods to decrease its prevalence is critical for industry participants and academic researchers. It has been demonstrated that coccidiosis can be effectively controlled and managed by employing anticoccidial chemical compounds. However, as a result of their extensive use, anticoccidial drug resistance in Eimeria species has raised concerns. Phytochemical/herbal medicines (Artemisia annua, Bidens pilosa, and garlic) seem to be a promising strategy for preventing coccidiosis, in accordance with the "anticoccidial chemical-free" standards. The impact of herbal supplements on poultry coccidiosis is based on the reduction of oocyst output by preventing the proliferation and growth of Eimeria species in chicken gastrointestinal tissues and lowering intestinal permeability via increased epithelial turnover. This review provides a thorough up-to-date assessment of the state of the art and technologies in the prevention and treatment of coccidiosis in chickens, including the most used phytochemical medications, their mode of action, and the applicable legal framework in the European Union.

Coccidiosis infection and growth performance of broilers in experimental trials: insights from a meta-analysis including modulating factors

An infection by protozoa Eimeria spp. can cause coccidiosis, which negatively affects broiler chicken performance and causes economic and production losses. To understand the effect of coccidiosis on broilers' performance, we evaluated the independent variables and their interactions on the severity of coccidiosis in broilers that cause variation (Δ) of average daily feed intake (ADFI), average daily gain (ADG), and gain per feed (G:F) of broiler chicks using a meta-analysis approach. A database of 55 papers describing 63 experiments was gathered; broilers were challenged by Eimeria species (E. acervulina, E. maxima, E. tenella, and mixed) and at least 2 variables among ADFI, ADG, and feed conversion ratio (FCR) were studied. The variation induced by the challenge was calculated relative to the control group of each experiment. The indirect factors evaluated were days postinfection (DPI), Eimeria type and dose, infection age (IA), bird's mean age in the analyzed period, genetic line, sex, and whether they were raised in a cage or a pen. Graphical, correlation, and variance analyses were performed to evaluate the form of the responses. Then, a linear plateau model was adjusted for each response variable as a function of DPI to determine the consequences of the disease on the variation of performance over time after infection. The impact of the infection challenge on the variation of performance vs. nonchallenge broilers was only impacted by DPI (P < 0.05). The adjustment of the data with the linear plateau model allows us to determine the host response to the coccidiosis disease at different stages. At 5 DPI (acute phase), ΔADFI, ΔADG, ΔG:F were of -19.0; -39.8, and -25.5, respectively. After almost 13 DPI birds achieved the recovery phase for all variables with Δ varying from -19 to -3.75% for ADFI, from -39.8 to -10.5% for ADG, and from -25.5 to -7.24% for G:F. The Eimeria impact was higher in ADG than ADFI in all periods due to Eimeria aggressive action form causing lesions in gut epithelial reducing the use of nutrients and energy. The results can be used as a quantitative approach to determine the consequences of Eimeria spp. on broiler performance.

Epidemiology and identification of Eimeria species affecting poultry in East Gojjam Zone, North West Ethiopia.

Despite the expansion of modernized poultry farming in Ethiopia, the presence of high prevalence of Eimeria species is the bottleneck in the sector causing high morbidity and mortality rate in poultry. The objectives of this study were to estimate the prevalence and identify Eimeria species and investigate the major risk factors. A cross-sectional study was conducted from November 2019 to April 2020 in East Gojjam Zone, North West Ethiopia. A total of 384 chickens were used. Both floatation and McMaster coprological techniques were employed. Univariate and multinomial logistic regression was used to calculate the odds ratio for the associated risk factors. Analysis of variance was used to analyse differences in Eimeria oocyst counts among the groups. Overall prevalence of Eimeria species in poultry from the study area was 26.5%. Age (OR=0.25, p=0.001), management system (OR=12.44, p=0.001) and production system (OR=0.37, p=0.001) were found significantly (p<0.05) associated with the risk of Eimeria species in poultry. The mean Eimeria oocyst count was significantly different by age and management system (F=6.526, p=0.002), (F=5.369, p=0.005), respectively. The mean Eimeria oocyst count was significantly greater in 6-12 weeks (p=0.004) and <6 weeks of age (p=0.025). A total of 6 Eimeria species were identified. Eimeria tenella (46.07%), Eimeria necatrix (24.5%) and Eimeria acervulina (8.82%) were the most common Eimeria species encountered. The prevalence of Eimeria species was higher in poultry in North West Ethiopia. Therefore, tailor-made intervention is required to mitigate risk factors and reduce the prevalence of Eimeria species in poultry from the study area.

Eimeria erythrorhynchosi n. sp. (Apicomplexa: Eimeriidae) from the American white pelican Pelecanus erythrorhynchos Gmelin, 1789 (Pelecaniformes: Pelecanidae) in Toluca, Mexico

A new coccidian species, Eimeria erythrorhynchosi n. sp. (Apicomplexa: Eimeriidae) collected from the American white pelican Pelecanus erythrorhynchos Gmelin 1789, is reported from Toluca city, Mexico. Sporulated oocysts of the new species are ovoidal, 17.5–19.6 × 14.1−14.8 (18.5 × 14.5) μm, with a length/width (L/W) ratio of 1.0–1.5 (1.25); micropyle present, robust and protruding, W: 4.5–5.5, and polar body (1 or 2), L × W: 2.5 (2–3), associated with the micropyle; oocyst residuum absent. Sporocysts are ovoidal, 10.1–11.2 × 6.4−7.1 (10.7 × 6.8) μm, with a length/width (L/W) ratio of 1.0–1.6 (1.3); Stieda and sub-Stieda bodies are both present, but para-Stieda body is absent; sporocyst residuum of diffuse granules. Sporozoites are vermiform, 9.6 × 2.4 μm. The new species is the seventh species of Eimeria recorded in Pelecaniformes and the third in Pelecanidae.

Characterization of the novel glucose-methanol-choline (GMC) oxidoreductase EnOXIO1 in Eimeria necatrix

Eimeria species are intracellular obligate parasites, among the most common pathogens affecting the intensive poultry industry. Oxidoreductases are members of a class of proteins with redox activity and are widely found in apicomplexan protozoans. However, there have been few reports related to Eimeria species. In this study, total RNA was extracted from the gametocytes of E. necatrix Yangzhou strain to amplify the EnOXIO1 gene using reverse-transcription polymerase chain reaction. After cloning and sequence analysis, the prokaryotic expression vector pET-28a(+)-EnOXIO1 was constructed and transformed into Escherichia coli BL21(DE3), and the recombinant protein rEnOXIO1 was expressed by induction with isopropyl ß-D-1-thiogalactopyranoside. The full length EnOXIO1 gene was 2535 bp encoding 844 amino acids, and the EnOXIO1 protein had a molecular weight of about 100 kDa and was mainly expressed in inclusion bodies. Western blot analysis indicated that the rEnOXIO1 protein had good antigenicity and cross-reactivity and was specifically recognized by a 6 ×HIS labeled monoclonal antibody, mouse anti-recombinant protein polyclonal antibody, and recovery serum from chickens infected with E. necatrix, E. acervulina, and E. tenella sporulated oocysts. The results of laser confocal immunofluorescence localization showed that the EnOXIO1 protein was mainly located on the wall-forming bodies in gametocytes and played an important role in the formation of the oocyst wall. Quantitative PCR analysis revealed that transcript levels of EnOXIO1 were highest in the gametocyte stage. Protein expression levels of EnOXIO1 were higher in the gametocyte stage than in other developmental stages according to western blot analysis. Vaccination of chickens against E. necatrix was achieved with recombinant protein rEnOXIO1, which triggered humoral immunity and antibody production, increased average body weight gain, reduced oocyst output and alleviated lesions after E. necatrix infection. The highest ACI value (172.36) was observed in chickens that received 200 μg rEnOXIO1 compared with other immunization groups.

Morphological and molecular characteristics of a single oocyst for the identification of Eimeria species in domestic rabbits (Oryctolagus cuniculus f. domesticus)

Coccidiosis caused by Eimeria is one of the most common diseases in domestic rabbits (Oryctolagus cuniculus f. domesticus), with 11 Eimeria species in domestic rabbits recognized internationally. To identify Eimeria species more accurately, a method based on the molecular characteristics of a single oocyst with multiple gene loci was established by combining morphological and molecular biology. The results showed that the total infection rate of Eimeria in domestic rabbits was 44.2 % (152/344). Ten Eimeria species were identified in domestic rabbits based on morphological characteristics, namely Eimeria vejdovskyi (39.5 %, 136/344), E. magna (18.0 %, 62/344), E. perforans (17.4 %, 60/344), E. intestinalis (12.5 %, 43/344), E. media (11.9 %, 41/344), E. coecicola (4.4 %, 15/344), E. irresidua (3.8 %, 13/344), E. exigua (2.6 %, 9/344), E. stiedai (2.3 %, 8/344), and E. piriformis (1.5 %, 5/344). The molecular biological identification of Eimeria in domestic rabbits was conducted through single oocyst selection and nested polymerase chain reaction amplification with multiple gene loci. We obtained the sequences of the 18S rRNA, ITS-1 and COI gene loci of E. magna, E. perforans, E. vejdovskyi, E. media, E. intestinalis, and E. coecicola. The results showed that the molecular biology and morphological identification results of single oocysts were consistent and could be used for the molecular identification of Eimeria at the single oocyst level. This study provides an efficient tool for identification of Eimeria in domestic rabbits and the population genetic study of Eimeria in domestic rabbits.

Identification of Eimeria tenella sporozoite immunodominant mimotopes by random phage-display peptide libraries-a proof of concept study.

Coccidiosis, caused by parasites of numerous Eimeria species, has long been recognized as an economically significant disease in the chicken industry worldwide. The rise of anti-coccidian resistance has driven a search for other parasite management techniques. Recombinant antigen vaccination presents a highly feasible alternative. Properly identifying antigens that might trigger a potent immune response is one of the major obstacles to creating a viable genetically modified vaccine. This study evaluated a reverse immunology approach for the identification of B-cell epitopes. Antisera from rabbits and hens inoculated with whole-sporozoites of E. tenella were used to identify Western blot antigens. The rabbit IgG fraction from the anti-sporozoite serum exhibited the highest reactogenicity; consequently, it was purified and utilized to screen two random Phage-display peptide libraries (12 mer and c7c mer). After three panning rounds, 20 clones from each library were randomly selected, their nucleotide sequences acquired, and their reactivity to anti-sporozoite E. tenella serum assessed. The selected peptide clones inferred amino acid sequences matched numerous E. tenella proteins. The extracellular domain of the epidermal growth factor-like (EGF-like) repeats, and the thrombospondin type-I (TSP-1) repeats of E. tenella micronemal protein 4 (EtMIC4) matched with the c7c mer selected clones CNTGSPYEC (2/20) and CMSTGLSSC (1/20) respectively. The clone CSISSLTHC that matched with a conserved hypothetical protein of E. tenella was widely selected (3/20). Selected clones from the 12-mer phage display library AGHTTQFNSKTT (7/20), GPNSAFWAGSER (2/20) and HFAYWWNGVRGP (8/20) showed similarities with a cullin homolog, elongation factor-2 and beta-dynein chain a putative E. tenella protein, respectively. Four immunodominant clones were previously selected and used to immunize rabbits. By ELISA and Western blot, all rabbit anti-clone serums detected E. tenella native antigens. Thus, selected phagotopes contained recombinant E. tenella antigen peptides. Using antibodies against E. tenella sporozoites, this study demonstrated the feasibility of screening Phage-display random peptide libraries for true immunotopes. In addition, this study looked at an approach for finding novel candidates that could be used as an E. tenella recombinant epitope-based vaccine.

Protective efficacy of recombinant proteins AMA1 and IMP1 in rabbits infected with Eimeria intestinalis

Eimeria intestinalis is one of the most pathogenic rabbit coccidia species causing severe intestinal damage and increased risk of secondary infection from opportunistic pathogens, which results in huge economic losses to the rabbit industry. Anticoccidial drugs are currently the main method to control coccidiosis; however, increasing resistance and drug residues have fueled research on anticoccidial vaccines. Apical membrane antigen 1 (AMA1) and immune mapped protein 1 (IMP1), as surface proteins, are associated with host invasion and might have the potential as candidate vaccine antigens. In the present study, recombinant IMP1 (rEiIMP1) and AMA1 (rEiAMA1) from E. intestinalis were expressed using Escherichia coli BL21. The immunoreactivity and immunoprotective effects of rEiIMP1 and rEiAMA1 were then analyzed. Fifty rabbits were grouped randomly (n = 10 per group): The unimmunized-unchallenged control group (sterilized phosphate-buffered saline (PBS)), the unimmunized-challenged control group (sterilized PBS), the vector protein-challenged control group (100 μg of pET-32a vector protein per rabbit), the rEiIMP1 immunized group (100 μg of rEiIMP1 per rabbit), and the rEiAMA1 immunized group (100 μg of rEiAMA1 per rabbit). After two immunizations, the rabbits were challenged with homologous oocysts (except for the unimmunized-unchallenged group). Serum specific antibody levels were assessed weekly throughout the experimental period; and the levels of different cytokines in the serum before the challenge were detected. The clinical symptoms, oocysts output, weight gain, feed conversion ratio (FCR), and lesion scores were recorded after experimental infection, and the anticoccidial indexes (ACIs) were calculated. The results showed that both rEiIMP1 and rEiAMA1 had good immunoreactivity. Rabbits immunized with rEiIMP1 and rEiAMA1 displayed 66.74 % and 63.14 % oocyst reduction, respective land 81.79 % and 78.87 % body weight gain, respectively. The rEiIMP1 and rEiAMA1 groups had lower FCRs (3.77:1 and 4.06:1, respectively) and lesion scores (P = 0.00). The rEiIMP1 and rEiAMA1 showed moderate effects, with an ACI of 152.09 and 147.17, respectively. Immunization induced high levels of anti-rEiIMP1 and -rEiAMA1 antibodies. Rabbits immunized with rEiIMP1 and rEiAMA1 displayed significantly increased interleukin (IL)− 2 (P = 0.00), interferon gamma (IFN)− γ (P = 0.00), and IL− 4 (P = 0.00) levels. Therefore, this study provided potential candidate vaccine antigens for E. intestinalis.

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

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. 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. 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. 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.

Incidence and therapeutic management of Eimeria species infection in Murrah buffalo calves

The present study was undertaken to examine the incidence of Eimeria species infection in Murrah buffalo calves and evaluate the therapeutic management of the disease. In November 2019, 9 to 10 buffaloes calves up to 3 months of age showed symptoms of diarrhoea, loss of appetite, and poor weight gain at an organised dairy farm. Based on history, faecal samples were collected from 20 buffaloes calves and subjected to standard qualitative coprological examination and quantitative analysis using a modified McMaster technique. Out of 20 faecal samples, 90% were positive for coccidia oocysts. Among positive samples, 11% were heavily infected and 16% had a moderate level of infection of Eimeria spp. Six species of Eimeria were identified, E. bareillyi was the most often encountered species, accounting for 37.39%, followed by E. bovis (22.76%), E. zuernii (14.63%), E. ellipsoidalis (13.82%), E. canadensis (13.82%), and E. cylindrica (6.50%). Blood examination of mild and severely affected calves revealed a decrease in the mean values of hemoglobulin, total erythrocyte count, total leucocyte count, platelets and an increase in monocytes as compared with healthy calves. The affected calves were treated with combination of Sulphadiazine and Trimethoprim (Biotrim IV injection) along with supportive treatment for five successive days. All the affected animals responded well to the treatment and showed marked reduction in clinical symptoms as well as oocyst output.

Epidemiology and Risk Factors of Coccidiosis in Broilers: Investigating the Prevalence, Distribution, and Risk Factors Associated with Coccidiosis Outbreaks in Broiler Production

Background: Coccidiosis is highly prevalent parasitic disease posing significant challenges to broiler production worldwide.&#x0D; Objectives: This study aimed to investigate the prevalence, distribution, and risk factors associated with coccidiosis outbreaks in broiler production systems.&#x0D; Methods: A cross-sectional study design was employed, and a total of 15 flocks, comprising 16,300 birds, were examined.&#x0D; Results: The overall prevalence rate of coccidiosis was found to be 66.7%. Among individual farms, Farm 1 reported a prevalence rate of 60.0%, Farm 2 had a rate of 50.0%, and Farm 3 exhibited the highest incidence at 83.3%. The specific species distribution within the positive flocks varied across farms. Farm 1 showed a mixed infection involving Eimeria tenella and Eimeria maxima, while Eimeria necatrix and Eimeria acervulina were found in Farm 2. Farm 3 exhibited a more diverse species distribution, including Eimeria necatrix, Eimeria tenella, Eimeria acervulina, and Eimeria maxima. Furthermore, this study identified several risk factors associated with coccidiosis outbreaks in broiler production. The analysis revealed that larger farm size, inadequate biosecurity measures, and poor feed quality were significantly associated with an increased risk of coccidiosis outbreaks.&#x0D; Conclusion: These findings provide valuable insights into the epidemiology of coccidiosis in broiler production and the factors influencing its occurrence. By targeting the prevalent Eimeria species and addressing the identified risk factors, poultry producers can minimize the impact of coccidiosis on broiler health, welfare, and economic performance.

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

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

Effect of tunable femtosecond laser pulses on the sporulation of Eimeria species oocysts of chickens: An in vitro study

Parasitic Eimeria species infection is a severe disease in the poultry industry and a major cause of coccidiosis and disease-engendered financial losses. Increasing resistance to drug-based treatment presents a significant problem, as does the use of disinfectants to sanitize poultry farms, because of increasing oocyst resistance to chemical interventions and contamination of poultry products. Reducing the rate of sporulation of Eimeria species oocysts can minimize the spread of the disease. Therefore, we studied the effect of femtosecond laser pulses on oocysts sporulation rate with the goal of developing an effective light-based treatment for coccidiosis, as an alternative to pharmaceutical intervention. Eimeria species oocysts were harvested from the intestines of infected broiler chickens. Unsporulated oocysts obtained were distributed over 96 well plates and irradiated with pulsed femtosecond laser at various wavelengths and exposure times. Irradiation of the oocysts with pulsed 360 to 390 nm femtosecond laser significantly reduced their sporulation rate and altered their morphology compared to untreated controls. Compared to other wavelengths within the 360 to 390 nm spectra, treatment with pulsed 370 nm wavelength was most effective in decreasing oocyst sporulation rate, reducing sporulation to 17%, 15%, and 11%, after 5-, 10-, and 15-min exposure times, respectively. In contrast, irradiation with infrared 720 nm and 800 nm wavelengths had no significant effect on oocysts sporulation rate. These findings clearly show that irradiation with a pulsed 370 nm femtosecond laser effectively reduces Eimeria species oocyst sporulation and that it is possible to develop an effective light-based treatment for coccidiosis.

Amplicon sequencing allows differential quantification of closely related parasite species: an example from rodent Coccidia (Eimeria)

BackgroundQuantifying infection intensity is a common goal in parasitological studies. We have previously shown that the amount of parasite DNA in faecal samples can be a biologically meaningful measure of infection intensity, even if it does not agree well with complementary counts of transmission stages (oocysts in the case of Coccidia). Parasite DNA can be quantified at relatively high throughput using quantitative polymerase chain reaction (qPCR), but amplification needs a high specificity and does not simultaneously distinguish between parasite species. Counting of amplified sequence variants (ASVs) from high-throughput marker gene sequencing using a relatively universal primer pair has the potential to distinguish between closely related co-infecting taxa and to uncover the community diversity, thus being both more specific and more open-ended.MethodsWe here compare qPCR to the sequencing-based amplification using standard PCR and a microfluidics-based PCR to quantify the unicellular parasite Eimeria in experimentally infected mice. We use multiple amplicons to differentially quantify Eimeria spp. in a natural house mouse population.ResultsWe show that sequencing-based quantification has high accuracy. Using a combination of phylogenetic analysis and the co-occurrence network, we distinguish three Eimeria species in naturally infected mice based on multiple marker regions and genes. We investigate geographical and host-related effects on Eimeria spp. community composition and find, as expected, prevalence to be largely explained by sampling locality (farm). Controlling for this effect, the novel approach allowed us to find body condition of mice to be negatively associated with Eimeria spp. abundance.ConclusionsWe conclude that amplicon sequencing provides the underused potential for species distinction and simultaneous quantification of parasites in faecal material. The method allowed us to detect a negative effect of Eimeria infection on the body condition of mice in the natural environment.Graphical

Prevalence of Avian Coccidiosis and Identification of Eimeria spp in Local Broilers and Chickens in Lafia Modern Market, Nassarawa State, Nigeria

Coccidiosis is an important enteric parasitic protozoan disease of poultry. The disease is usually associated with high economic losses to poultry farmers worldwide. This study was conducted to determine the prevalence of coccidiosis, identify the species of circulating Eimeria species, determine the prevalence of Eimeria in respect to breed and gender and determine the deviation of infected birds from normal range of birds slaughtered in Lafia modern market, Nasarawa State, Nigeria. Out of the 204 examined chickens, 36.3% (74/204) of them harbored different Eimeria species. A total of 32.6% (44/74) male and 43.5 (30/74) female were infected. Statistically, no significant difference p&gt;0.05 (χ2=1.559, p=0.2118) was noted between the male and female. Among the local breed, 40.2% (49/74) was infected and for the broilers, 30.5% (25/74) was positive for coccidial infection. Again, there was no statistically significant difference p&gt;0.05 (χ2=1.324, p=0.25) between the local and broiler breeds. In the attempt made to identify the prevalent species of coccidian in the study area, five Eimerian species were identified, namely; Eimeria acervulina, Eimeria brunette, Eimeria mitis, Eimeria necatrix and Eimeria tenella with the prevalence of 14.22%, 5.88%, 1.47%, 6.86%, and 7.84% respectively. Results obtained from this study shows that coccidiosis is an important disease of chickens in the study area and further strategy needs to be implemented to reduce the infection rate of coccidiosis.

Isospora speciosae n. sp. (Apicomplexa: Eimeriidae) from the black-polled yellowthroat Geothlypis speciosa Sclater, 1859 (Passeriformes: Parulidae) in Ciénegas del Lerma, Mexico.

A new coccidian species, Isospora speciosae n. sp. (Apicomplexa: Eimeriidae) collected from the black-polled yellowthroat Geothlypis speciosa Sclater, is reported from the marsh Ciénegas del Lerma Natural Protected Area, Mexico. Sporulated oöcysts of the new species are subespherical to ovoidal, 24-26 × 21-23 (25.7 × 22.2) μm, with a length/width (L/W) ratio of 1.1; one or two polar granules are present, but micropyle and oöcyst residuum are absent. Sporocyts are ovoidal, 17-19 × 9-11 (18.7 × 10.2) μm, with a L/W ratio of 1.8; Stieda and sub-Stieda bodies are both present, but para-Stieda body is absent; sporocyst residuum compact. The new species is the sixth species of Isospora recorded in a bird of the family Parulidae in the New World.

Effect of Nutrients and Herbal Products on Immune Status of Broilers, Experimentally Infected with Mixed Eimeria Species

The present study was conducted to seek herbal alternatives and nutritional modulation to solve the problem of coccidiosis, a deadly protozoan disease of broilers. For the purpose, an experiment of 35 days’ duration (days 0-21 pre-infection period and days 22-35 post-infection period) was conducted to test the efficacy of herbal/natural products/dietary nutrients for the management of mixed coccidial infection prevalent under field conditions. For this purpose wheat grass juice, chelated mineral supplement, herbal anticoccidial and seeds of Embelia ribes without and with sodium bicarbonate, organic chromium, herbal vitamin E-Selenium without and with organic chromium were used to study the immunological parameters of un-infected and infected broilers with 60,000 sporulated oocysts of mixed Eimeria species. Results of the experiment revealed that LST in terms of delta optical density of T- and B-cells was maximum in wheat grass juice while ELISA titre against Newcastle Disease was maximum in herbal vitamin E-Selenium group. All the types of dietary modulation were found to be effective up to different extent in the management of poultry coccidiosis. Further research should focus on micro-array and proteomic technologies to solve the problem.

First report of Eimeria myoxi in the garden dormouse (Eliomys quercinus Linnaeus, 1766) from Doñana Natural Area (Andalusia, SW Spain).

This work reports for the first time the presence and molecular characterization of Eimeria myoxi in the garden dormouse (Eliomys quercinus) from the Doñana Natural Area (Andalusia, SW Spain). Fresh faecal samples were collected from a total of 28 garden dormice, which were caught following current guidelines for the ethical use of animals in research, and processing by a standard flotation technique with saturated saline solution. Then, wet drops were examined microscopically, and the number of oocysts was semi-quantified. Eimeria oocysts were observed in 16 of the 28 (57.1%) faecal samples, showing most of them a very low number of oocysts (≤1 oocyst per microscopic field × 400). The unsporulated oocysts visualized in 16 faecal samples were subspherical and of length 19.2±1.2μm and width 17.4±1.1μm, being morphologically compatible with E. myoxi. This finding was supported by molecular analysis of the small subunit ribosomal RNA (SSU-rRNA) gene, identifying the same species in 22 of the 28 (78.6%) dormice, including 15 samples in which oocyst size was compatible with E. myoxi. Moreover, the subsequent analyses of the apicoplast open reading frame 470 (ORF470) and the mitochondrial cytochrome c oxidase subunit I (COI) genes confirmed the molecular identification of the isolates as E. myoxi. The phylogeny analyses were consistent with previous phylogenetic studies and support the existence of three lineages of rodent-infecting Eimeria species.