Sporulated Oocysts Research Articles

In vitro studies for antiparasitic activities of Punica granatum extract.

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

Label-free quantitative detection and comparative analysis of lysine acetylation during the different life stages of Eimeria tenella.

Proteome-wide lysine acetylation has been documented in apicomplexan parasite Toxoplasma gondii and Plasmodium falciparum. Here, we conducted the first lysine acetylome in unsporulated oocysts (USO), sporulated 7 h oocysts (SO 7h), sporulated oocysts (SO), sporozoites (S), and the second generation merozoites (SMG) of Eimeria tenella through a 4D label-free quantitative technique. Altogether, 8532 lysine acetylation sites on 2325 proteins were identified in E. tenella, among which 5445 sites on 1493 proteins were quantified. In addition, 557, 339, 478, 248, 241, and 424 differentially expressed proteins were identified in the comparisons SO7h vs USO, SO vs SO7h, SO vs USO, S vs SO, SMG vs S, and USO vs SMG, respectively. The bioinformatics analysis of the acetylome showed that the lysine acetylation is widespread on proteins of diverse functions. Moreover, the dynamic changes of lysine acetylome among E. tenella different life stages revealed significant regulation during the whole process of E. tenella growth and stage conversion. This study provides a beginning for the investigation of the regulate role of lysine acetylation in E. tenella and may provide new strategies for anticoccidiosis drug and vaccine development. Raw data are publicly available at iProX with the data set identifier PXD040368.

Tratamento de isosporose em felinos com ponazuril: Relato de caso

The occurrence of gastrointestinal parasites in cats is reported with great frequency, especially when it comes to animals with outdoor access, or living in overcrowded environments, such as catteries and shelters. Infection byIsospora spp in cats begins with the ingestion of sporulated oocysts in the environment or tissues from other infected intermediate vertebrate hosts. Diagnosis is based on the detection of oocysts in cat feces and clinical signs. The recommended treatment for coccidiosis in cats is sulfonamides. Other drugs have been studied as secondary lines of treatment, which is the case of ponazuril and toltrazuril, which have been used in animals to treat isosporosis, toxoplasmosis, neosporosis and equine protozoal meningoencephalitis. There are anecdotal reports of the use of ponazuril in treating Isospora spp. In this study, 21 cats were treated with ponazuril, which proved effective in controlling the disease, since 100% of the cats treated were negative in the coproparasitological examination after the three days of treatment with a dose of 50 mg/kg SID. Furthermore, the animals had considerable improvement in stool consistency after the dosing period.

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.

English

Essential oils are potent medicinal substances which can control multiple diseases by their antioxidant actions. In current research, anticoccidial potential of the Amomum subulatum essential oil was estimated in broiler chicks. 270 broiler chicks were divided equally into 6 groups, each having 3 replicates. All the groups, except infected nonmedicated control group, were given sporulated oocysts of mixed Eimeria spp. First three groups received essential oil of A. subulatum @ 1%, 2% and 3% (v/w) in the feed and their activities was evaluated in comparison to the Symocox® (Toltrazuril) treated groups and infected nonmedicated groups. Feed conversion ratio, feed intake, weight gain, rates of mortality, organ weight ratios, fecal scores, lesion scores and oocyst per gram of feces were taken as the parameters of the study. Blood parameters and serum metabolites were also evaluated. A. subulatum essential oil @ 3% concentration had the best FCR, survival rates, and feed intakes. A. subulatum essential oils also significantly reduced lesion lowered oocyst counts and improved the fecal scores. Hematological and serum profiles were also improved by the 3% essential oil of A. subulatum in the feed. This research concluded that the A. subulatum essential oil @ 3% concentration can be used for the treatment of avian coccidiosis however formulation of dose requires further studies on broiler chicks.

Actividad anticoccidial in vitro de microencapsulados de aceites esenciales frente a Eimeria spp

The aimof this research was to evaluate the in vitro anticoccidial activity of microencapsulated essential oils from Lepechinia heteromorpha(Briq.) Epling and Salvia sagittataRuiz & Pav., against non-attenuated sporulated oocysts of Eimeriasppof broiler chickens. The treatments were the concentrations of the microencapsulated in 2.5; 5; 10; 20;30 and 50 mg/ml, 2.5% potassium dichromate negative control group and 4% formaldehyde positive control group. The destruction of oocysts in liquid medium was evaluated by microscopic counting and the results were verified after 24 and 48 h of incubation at 28 °C. Data were analyzed by analysis of variance, using SPSS version 25 software and Tukey's test. The results showed highly significant statistical differences between the microencapsulated, as well as in their concentrations, registering the greatest destruction of sporulated oocysts by the microencapsulated essential oil of Salvia sagittataRuiz & Pav. at a concentration of 50 mg/ml at 48 h of incubation, reaching83.5% destruction of oocysts, this value being similar to 4% formaldehyde. Concluding that the microencapsulated essential oils studied destroy up to 83.5% of the sporulated oocysts of Eimeriaspp.

In Vitro: The Effects of the Anticoccidial Activities of Calotropis procera Leaf Extracts on Eimeria stiedae Oocysts Isolated from Rabbits.

Hepatic coccidiosis is an infectious and mortal disease that causes global economic losses in rabbits. The research aimed to assess the efficacy of Calotropis procure leaf extracts on the inhibition of Eimeria stiedae oocysts and to determine the optimal dosage for suppressing the parasite's infective phase. In this experiment, oocyst samples per milliliter were tested, and 6-well plates (2 mL) of 2.5% potassium dichromate solution containing 102 non-sporulated oocysts on Calotropis procera leaf extracts were exposed after 24, 48, 72, and 96 h, and the treatments were as follows: a nontreated control, 25%, 50%, 100%, and 150% of C. procera for oocyst activities. In addition, amprolium was utilized as a reference drug. The Calotropis procera was analyzed by GC-Mass, and results showed that the botanical extract contained 9 chemical components that were able to inhibit the oocysts of E. stiedae at 100% and 150% concentrations by about 78% and 93%, respectively. In general, an increase in the incubation period and a greater dose resulted in a decrease in the inhibition rate. The results showed that C. procera has an effective ability, inhibitory potential, and protective effect on the coccidian oocyst sporulation of E. stiedae. It can be used in the disinfection and sterilization of poultry and rabbit houses to get rid of Eimeria oocysts.

The In Vitro Anticoccidial Activity of Some Herbal Extracts against Eimeria spp. Oocysts Isolated from Piglets

Coccidiosis in pigs seldom results in important economic losses. However, it can influence growth rates in weaners and it is an important hygiene indicator in swine farms. Certain herbs, along with their extracts, have been used over the course of history in traditional medicine. The aim of this study was to evaluate the in vitro anticoccidial effects of Allium sativum L. (garlic), Artemisia absinthium L. (wormwood), Coriandrum sativum L. (coriander), Cucurbita pepo L. (pumpkin), Satureja hortensis L. (summer savory), and Calendula officinalis L. (marigold) against Eimeria suis and Eimeria debliecki oocysts. The stock solution of oocysts (58% E. suis + 42% E. debliecki) was incubated for three days, before adding the tested solutions. The unsporulated Eimeria spp. oocysts were then placed in a 3 mL well plate and incubated for 96 h at 27 °C, in a suspension containing serial dilutions of alcoholic plant extracts (5%, 2.5%, 1.25%, 0.625%, and 0.312%). The percentage of sporulated and destroyed oocysts was recorded every 24 h for 96 h. All extracts had a good in vitro anticoccidial effect against oocysts of Eimeria spp. compared with the control groups. Oocyst sporulation was significantly (p = 0.05) inhibited by the 5% marigold extract (3.6% sporulated oocysts). The same extract had the highest lytic effect on oocysts (65.2% destroyed oocysts). Our results prove that the most effective alcoholic plant extract is the marigold extract, followed, in order of efficiency, by the wormwood, coriander, garlic, pumpkin, and summer savory extracts. This study represents a preliminary contribution for establishing a new generation of natural disinfectants aimed at destroying Eimeria oocysts in the context of swine contamination.

Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenella

Eimeria tenella, an intestinal parasite, has brought huge economic losses to the poultry industry. The prevalence and severity of the development of drug resistance has increased the challenge of coccidiosis control. We previously identified the enolase 2 of E. tenella (EtENO2) was differentially expressed in drug-sensitive (DS) and drug-resistant strains using RNA-seq. In this study, the expression of EtENO2 in diclazuril-resistant (DZR), maduramicin-resistant (MRR), and salinomycin-resistant (SMR) strains was analyzed by quantitative real-time PCR (qRT-PCR) and western blots. EtENO2 was highly expressed in several drug-resistant strains compared with the DS strain. The qRT-PCR showed that the transcription level of EtENO2 in the field-isolated resistant strains was upregulated compared with the DS strain. The enzyme activity results indicated that the catalytic activity of EtENO2 in the drug-resistant strains was higher than in the DS strain. In addition, qRT-PCR and western blots showed that the expression level of EtENO2 was higher in second generation merozoites (SM) and unsporulated oocysts (UO) than that in sporozoites (SZ) and sporulated oocysts (SO). Immunofluorescence localization revealed that EtENO2 was distributed throughout SZ and SM and on the surface of the parasites. After the SZ invasion DF-1 cells, it was also observed on the parasitophorous vacuole membrane. Our secretion experiments found that EtENO2 could be secreted outside the SZ. This study indicated that EtENO2 might be related to the interaction between E. tenella and host cells and be involved in the development of E. tenella resistance to some anticoccidial drugs.

The Anticoccidial Effect of Alcoholic Vitis vinifera Leaf Extracts on Eimeria papillate Oocysts Isolated in Mice In Vitro and In Vivo.

Eimeria spp. causes eimeriosis in the guts of numerous domestic mammals and poultry, and the employment of medication and the effects of certain aspects of synthetic anticoccidials in the treatment of eimeriosis have given rise to the appearance of resistant parasites that require the search for alternate remedies. Natural products, which are safe and have no negative impact on the environment, may be utilized in the therapy of an enormous range of parasitic infections. This research aimed to assess the effectiveness of VVLE on the oocyst sporulation of an E. papillate infection in the mouse jejunum. In addition, obtaining the ideal concentration will interrupt the parasite's life cycle and limit infection. In vitro: Collected unsporulated oocysts (1 × 103) of E. papillata were given six different concentrations (w/v) of Vitis vinifera leaf extract (10, 25, 50, 100, 150, and 200 mg/mL) and toltrazuril (25 mg/mL), three replicates per group, whereas the control group received 2.5% potassium dichromate solution. In vivo: The mice were separated into six groups; the first and second groups did not receive infection, whilst the third, fourth, fifth, and sixth groups were each given 1 × 103 sporulated oocysts of E. papillate in the experiment. In addition, an oral dosage of 100 and 200 mg/kg VVLE were given to the fourth and fifth groups, while the sixth group was given toltrazuril at 25 mg/kg. On the fifth day, unpopulated oocysts were collected from each mouse separately. The incubation period and treatments had considerable impacts on the rate of sporulation. The infrared spectroscopy of V. vinifera extract revealed many expected active classes of chemical compounds. Further, the infection of mice with E. papillata caused an oocyst output of nearly 2 × 104 oocysts/g of faeces. VVLE significantly decreased the oocyst output to nearly 88%. In addition, we detected an inhibitory effect on the sporulation (%) and harm (%) of E. papillata oocysts in a dosage-dependent modality compared with the control group. Furthermore, they destroyed the oocyst morphology in terms of the shape, size, and quantity of sporocysts. The results indicate that grape vines have powerful activity as anticoccidials.

Investigation into the potential of using UV-treated sporulated oocysts of Eimeria tenella as a local solution to immunization of chickens against caecal coccidiosis.

In this study, we aim to evaluate the immune response of chickens to UV-treated sporulated oocysts as a means of protection against caecal coccidiosis caused by field strains of Eimeria tenella. Two groups of chicks were immunized using prepared UV-treated oocysts of E. tenella and challenged at day 20 post hatching. The first group was immunized only once at day 1 post hatching, the second group was immunized twice (day 1 and day 8 post hatching). Two non-immunized control groups were used: the first group was challenged with E. tenella, while the second group remained uninfected. The effectiveness of immunization on production and animal health was evaluated by the following criteria: body weight, feed conversion ratio, blood in faeces, mortality, lesion scores and oocyst output. The two immunized groups showed a significantly better performance in body weight, weight gain and lesion scores than the non-immunized group. However, all three groups performed significantly worse than the unchallenged group. The mortality of the non-immunized infected group was high (70%) while mortality in both immunized and unchallenged groups of chickens was significantly lower (range 2.2 to 4.4%) than the infected group (p < 0.05). The production of oocysts in faeces, post-infection, was significantly higher in the non-immunized group compared to the immunized group (p < 0.05) and both were significantly higher than the uninfected group (p < 0.05). In conclusion, immunization by prepared UV-irradiated oocysts is effective in stimulating at least a partial protective immunity in immunized chickens against caecal coccidiosis.

Biological activities of Persea americana: in vitro and in vivo studies

Natural resources emerge as an essential and alternative tool for controlling parasitic agents. The present study was carried out to evaluate in vitro and in vivo effect of Persea americana fruit extract (PAFE) on helminthiasis and coccidiosis. In vitro, anthelmintic activity was performed using PAFE against Allolobophora caliginose. PAFE elicited significant anthelmintic activity comparable to mebendazole with the lesser time taken to paralysis and death for 200 mg/mL. In vitro anticoccidial activity of PAFE, amprolium, phenol, DettolTM, and formalin were studied after incubation with unsporulated oocysts. PAFE was able to significantly inhibit oocyst sporulation in a dose-dependent manner. A total 35 male mice were divided into seven groups. At day 0, all groups except first and second groups were infected with 1×103 E. papillata. Group 1 served as control. Group 2 served as uninfected-treated one. Group 3 was considered an infected-untreated group. After 60 min of infection, groups 4, 5, and 6 were treated with PAFE. Group 7 was treated with amprolium. However, not at par with amprolium, PAFE showed a good anticoccidial activity adjudged based on a significant change in body weight gain, feed intake, oocysts output, and content of carbohydrates and protein. Therefore, PAFE exhibits powerful anthelmintic and anticoccidial effects.

Phytochemical Profile of the Methanolic Leaf Extract of Vernonia amygdalina Del. and In Vitro Anticoccidial Effect of Its Fractions against Eimeria tenella

Phytochemical screening of the crude methanolic leaf extract of Vernonia amygdalina was carried out. The extract and its fractions were tested for in vitro anticoccidial efficacy against the unsporulated oocysts of Eimeria tenella by the oocyst sporulation inhibition model in a 96-well microtitre plate. Phytochemical analysis of the extract revealed the presence of alkaloids, tannins, saponins, glycosides, and triterpenoids. Different concentrations of the extracts, including 250 mg/ml, 125 mg/ml, 62.5 mg/ml, 31.25 mg/ml, 15.625 mg/ml, and 7.8 mg/ml, were tested against freshly prepared unsporulated oocysts of E. tenella. Triplicate assays were conducted for each concentration of the extract to screen for anticoccidial activity. The methanol extract and fractions showed activity against the oocysts of E. tenella in a dose-dependent manner. The sporulation inhibition rate was determined as effective when the result showed more than 50% inhibition at each concentration level of treatment. Only the methanol extract and its butanol fraction at the lowest treatment concentration of 7.8 mg/ml showed an inhibition rate of 55% and 68%, respectively. All treatments: methanol extract (100%), hexane fraction (100%), butanol fraction (100%), and aqueous residue fraction (58%) were active at the highest concentration of 250 mg/ ml. It is concluded that the methanol leaf extract of Vernonia amygdalina and its butanol fraction had better oocyst sporulation inhibition activity against Eimeria tenella and may further be explored for their anticoccidial properties.

Serological and Molecular Detection of Toxoplasma Gondii among Pregnant Women Giving Birth at the University Of Gondar Specialized Hospital in Gondar Town, Northwest Ethiopia

Objective: Congenital toxoplasmosis can be presented when immunosuppressed women are reactivating of this infection acquired before pregnancy in addition to women recently exposed to this disease during pregnancy, which causes of fetal death, neonatal death or congenital defects. Women mainly get this infection by ingested improperly cooked or raw meat consisting of viable tissue cysts or by contaminated foods and water within sporulated oocysts or through the placenta. Thus, this study aimed to detect Toxoplasma gondii in pregnant women using serologically and molecularly at study area. Methods: A cross-sectional study was conducted from September 2019 to October 2020 by collecting a total of 200 umbilical cord blood and matching discharged placental tissue samples from systematic randomly selected pregnant women giving birth at University of Gondar specialized hospital. Latex agglutination test and nested polymerase chain reaction were used for the detections of this infection. Findings: The overall serological, first polymerase chain reaction and nested polymerase chain reaction findings of this study were 46%, 24% and 10%, respectively. The univariant analysis indicated that seropositivity of Toxoplasma gondii antibodies were also significantly influenced by eating raw meat, eating raw vegetables, drinking raw milk, presence of domestic cat contacts and source of water (P  0.05). In the case of multivariate analysis drinking raw milk and source of water were also significantly associated to the seropositivity of Toxoplasma gondii. Moreover, there were also fair concordant between latex agglutination within first polymerase reaction (Kappa: 0.290) andnested polymerase chain reaction tests of this parasite (Kappa: 0.231). Novelty: This nested polymerase chain reaction and the Toxo-latex agglutination tests had also fair agreements in pregnant women. Moreover, Consumption of raw meat, vegetables and milk; domestic cat contacts and source of water were the potential statically significant associated risk factors for Toxoplasma gondii seropositivity. Conclusions: Generally, these comparative tests confirmed the existence of this infection in pregnant women that played up a pool of public health risks for their embryo and infants. Thus, thoroughly cooking raw meats and vegetables; pasteurized raw milk; avoid domestic cat contacts and boiling of water will be the best prevention and control strategies of this infection. Further works on its genotyping of this pathogen will be also convinced. Keywords: Latex agglutination; Nested polymerase chain reaction; Placental tissue; Pregnant women; Toxoplasma gondii; Umbilical cord blood

Evaluation of the Anticoccidial Activity of Sheep Bile against Eimeria stiedae Oocysts and Sporozoites of Rabbits: An In Vitro Study.

Coccidiosis is one of the most common infectious diseases that causes digestive problems in rabbits, leading to global economic losses. This study was conducted to evaluate the effects of bile obtained from sheep gallbladder on the sporulation and morphology of Eimeria stiedae oocysts and sporozoites affecting rabbit liver cells and to determine the best concentration for sporulation inhibition. Sporulation inhibition per milliliter was measured in samples exposed to five concentrations of sheep bile (SB) in a 2.5% potassium dichromate solution: 12.5%, 25%, 50%, 75%, and 100% concentrations for oocysticidal activity and 125, 250, 500, 750, and 1000 μg/mL concentrations for antisporozoidal activity. A bioassay was performed to assess the in vitro anticoccidial activity of sheep bile against E. stiedae oocysts and sporozoite sporulation. In this assay, six-well plates with 5 mL of bile containing 1000 oocysts showed unsporulated oocysticidal activity after 48, 72, and 96 h and antisporozoidal activity after 12 and 24 h. A chemical assay was performed via infrared spectroscopy to investigate the presence of several anticipated active chemical compounds in sheep bile. Sheep bile was able to inhibit E. stiedae oocysts at 100% and 75% concentrations by about 91% and 81%, respectively. In addition, SB had the highest inhibition of E. stiedae sporozoite viability (92%) at a concentration of 1000 μg/mL and had the lowest inhibition of 8% at a concentration of 125 μg/mL. An increase in the incubation time and a higher dose generally increased the inhibition rate. The results showed that sheep gallbladder bile is effective due to its inhibitory potential and effect on the coccidian oocyst sporulation of E. stiedae. Further studies are needed to determine the precise active chemicals present in SB and their modes of action and application in vivo.

Comparative Effect of Allicin and Alcoholic Garlic Extract on the Morphology and Infectivity of Eimeria tenella Oocysts in Chickens.

Avian coccidiosis remains one of the major parasitic diseases that threaten the global poultry industry. Since prevention is superior to treatment, this study focuses on eliminating the infection outside the host. To determine their effect on the viability of Eimeria&amp;nbsp;tenella oocysts in vitro, allicin and alcoholic garlic extract, which are natural, less toxic, and inexpensive products, were compared to KOH 5% (chemical disinfectant) using an in vitro culture system. Three concentrations of allicin (45, 90, and 180 mg/mL) and alcoholic garlic extract (90, 180, and 360 mg/mL, were used. Subsequently, destructive and sporulation-inhibiting effects on Eimeria oocysts were detected using light and electron microscopy. Young chickens were infected with treated sporulated oocysts to determine their effect on infectivity. After 7 days pi, the percentage of excreted oocysts (oocyst shedding) was determined, and the chickens were slaughtered for histopathological examination of the cecal tissues. Under an electron microscope, allicin at a concentration of 180 mg/mL and alcoholic garlic extract at a concentration of 360 mg/mL demonstrate a high oocysticidal activity with severe destruction of the oocyst wall and the appearance of pores. In addition, both concentrations directly affected the infectivity of sporulated oocysts by reducing the shedding of oocysts and the pathological lesions of infected young chickens. We concluded that the ability of Allicin and alcoholic garlic extract to eliminate Eimeria oocysts makes them superior to chemical disinfectants as a disinfectant.

In vitro: Anti-coccidia activity ofCalotropis proceraleaf extract onEimeria papillataoocysts sporulation and sporozoite

AbstractNatural products play an important role as environmentally friendly agents that can be used against parasitic diseases. ManyEimeriaspecies cause eimeriosis in poultry. The negative effects of synthetic anti-coccidiosis medications necessitate the quest for alternative treatments derived from medicinal plants in the treatment of eimeriosis. The study was conducted to evaluate the effects ofCalotropis proceraleaf extract (CPLE) (Madar) on the sporulation ofEimeriaoocysts and sporozoites that affect mammalian jejunum and to obtain the best concentration for sporulation inhibition and infection prevention. Extracts were testedin vitroto prevent oocyst sporulation, wall deformity, and anti-sporozoite activity withEimeria papillata. The plant-chemical compounds analysis of CPLE some active compounds were shown as well as CPLEin vitroeffects at various concentrations (200, 100, 50, 25,12.5, and 6.25 mg/mL), while potassium dichromate solution 2.5% and Toltrazuril 25 mg/mL were administered as the control groups.C. proceraleaf extract showed the highest inhibitory percentage onE. papillataoocyst at 200 mg/mL of extract, approximately 91%. In addition, CPLE showed the sporozoite highest viability inhibitory percentage onE. papillataat 200 mg/mL of extract, approximately 88%, and the lowest efficacy was 5% at 6.25 mg/mL. Also, we noticed the deformation and destruction of the oocyst wall based on the concentration rate. Sporulation inhibition rate is significantly affected by incubation time and treatment concentration ratio. The results showed that Madar has an effective, inhibitory potential, and protective effect on coccidian oocyst sporulation and sporozoites ofE. papillata.

Label-free quantitative proteomic analysis of ethanamizuril-resistant versus -sensitive strains of Eimeria tenella

BackgroundAvian coccidiosis is an important parasitic disease that has serious adverse effects on the global poultry industry. The extensive use of anticoccidial drugs has resulted in an increase in drug resistance. Ethanamizuril (EZL) is a novel triazine with high anticoccidial activity.MethodsWe compared oocyst production and sporulation between EZL-sensitive (S) and EZL-resistant Eimeria tenella strains (R10 and R200) and used label-free quantitative proteomics to identify differentially expressed proteins (DEPs) between these strains.ResultsWe generated two EZL-resistant E. tenella strains: strain R10, which was induced using a constant dose of 10 mg EZL/kg poultry feed, and strain R200, which was generated by gradually increasing the EZL dosage to 200 mg EZL/kg poultry feed. With an increase in resistance, the total oocyst output decreased, but the percentage of sporulation did not change significantly. We identified a total of 7511 peptides and 1282 proteins, and found 152 DEPs in the R10 strain versus the S strain, 426 DEPs in the R200 strain versus the S strain and 494 DEPs in the R200 strain versus the R10 strain. When compared with the S strain, 86 DEPs were found to have consistent trends in both resistant strains. The DEPs were primarily involved in ATP and GTP binding, invasion, and membrane components. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the DEPs suggested that they are involved in transcription and translation processes. Protein–protein interaction network analysis of the 86 DEPs showed that 10 proteins were hubs in the functional interaction network (≥ 8 edges) and five of them were ribosomal proteins.ConclusionsThe results of the present study indicate that the resistance mechanisms of E. tenella against EZL might be related to the transcriptional and translational processes, especially in the factors that inhibit the growth of parasites. The DEPs found in this study provide new insights into the resistance mechanisms of E. tenella against EZL. Further research on these potential targets holds promise for new chemotherapeutic approaches for controlling E. tenella infections.Graphical

White-tailed eagle (Haliaeetus albicilla) as the definitive host of Sarcocystis lutrae in the Czech Republic.

The white-tailed eagle, Haliaeetus albicilla, has been involved in the life cycle of several Sarcocystis species as the intermediate and definitive host. To date, it has been supposed that the eagle might play the role as the definitive host for S. Lutrae, and, herein, we tried to elucidate it based on morphometric and molecular analyses. One out of two eagles harbored oocysts (17.0-17.4 × 11.3-11.9 μm) and sporocysts (11.3-12.3 × 8.3-9.3 μm) in the intestinal mucosa, whose sequences at 18S rRNA, 28S rRNA, ITS1, and cox1 showed similar identity (97.64-100%) to published sequences of S. lutrae from other hosts. The presence of sporulated oocysts in the lamina propria of villi confirms that S. lutrae truly infects the white-tailed eagle. The white-tailed eagle is confirmed as the definitive host of S. lutrae in the Czech Republic.

The Anticoccidial Effects of the Poultry Bile against Eimeria papillata Oocysts of Mice: In vitro Study

Background: Coccidiosis is a costly and wide-spread infectious disease in mammals that causes huge losses in the poultry industry worldwide. The present study was conducted to evaluate the potency of intact poultry gallbladder on the sporulation of Eimeria papillata oocysts of mice and to determine the best concentration for the activity of poultry bile on the inhibition of sporulation in oocysts. Methods: In this test, we used potassium dichromate solution (2.5%) as control group. Unpopulated oocysts were exposed (1 x 103/3 ml) to four concentrations of poultry bile (w/v; 10%, 25%, 50%, and 100%). It was incubated in petri dishes for 48, 72 and 96 hours at 25-29°C, with 60-80% humidity. The sporulation was examined under an inverted microscope at 40 x to confirm oocyst sporulation. Result: Showed through infrared spectroscopy of poultry bile that there are many expected active classes of chemical compounds. At a 100% concentration of poultry bile, it was able to inhibit the Eimeria papillata oocysts by about 98% in potassium dichromate solution. Also, it observed its ability to inhibit sporulation of oocysts in a dose-dependent manner at concentrations of 50%, 25%, and 10% of bile, the rate of inhibition was (71.7%, 33.11% and 19.88%), respectively. We found a direct relationship between inhibition and sporulation related to the increasing time period.