Effects of three kinds of Chinese herbs on growth performance, oocysts output and gut microbiota in growing lambs with coccidiosis.

Coccidiosis is a protozoan intestinal disease that reduces the production of the sheep industry and causes large economic losses for sheep. Although chemically synthesised drugs are routinely employed to treat coccidiosis in sheep, the anticoccidial drug resistance and drug residues in edible meat have prompted an urgent search for alternatives. Herein, the anticoccidial properties of diclazuril, a conventional anticoccidial drug, and Allium sativum, Houttuynia cordata and Portulaca oleracea were assessed. Forty 45-day-old lambs naturally infected with Eimeria spp. were selected and randomly divided into five groups. The results showed that the sheep treated for coccidiosis had considerably decreased average daily gain (ADG) during both administration and withdrawal of the drug compared to the control group. Furthermore, at days 14, 21, 28 and 35, respectively, the three herbs and diclazuril had similar anticoccidial effects, with lower oocysts per gram (OPG) than the control group. On day 78, OPG in the three herbal groups was significantly lower than in the diclazuril group. In addition, the abundance and composition of the gut microbiota were changed in sheep treated with the three herbs and diclazuril compared to the untreated sheep. Moreover, some intestinal microorganisms have a correlation with OPG and ADG when using Spearman correlation analysis. In summary, our results suggest that all three herbs produce anticoccidial effects similar to diclazuril and modulate the balance of gut microbiota in growing lambs.

Green synthesis of zinc oxide/Allium sativum nano-composite and its efficacy against murine cryptosporidiosis.

Cryptosporidiosis is a global health problem threats life of immunocompromised patients. Allium sativum (A. sativum) is one of the therapeutic options for cryptosporidiosis. This study develops green synthesized ZnO-NPs based on A. sativum extract, and assesses its therapeutic application in treating experimental cryptosporidiosis in immunosuppressed mice. FTIR, scanning electron microscopy, and zeta analyzer were used for characterization of bio ZnO-NPs. The morphology of prepared materials appeared as sponge with many pores on the whole surface that allows the feasibility of bio ZnO-NPs for different biological activities. Its structural analysis was highly stabilized with negative charge surface which indicated for well distribution into the parasite matrix. Twenty-five immunosuppressed Cryptosporidium parvum infected mice, classified into 5 groups were sacrificed at 21th day after infection with evaluation of parasitological, histopathological, oxidative, and proinflammatory biomarkers. Treated mice groups with 50 and 100 mg/kg of AS/ZnO-NPs showed a highly significant decline (79.9% and 83.23%, respectively) in the total number of expelled oocysts. Both doses revealed actual amelioration of the intestinal, hepatic, and pulmonary histopathological lesions. They also significantly produced an increase in GSH values and improved the changes in NO and MDA levels, and showed high anti-inflammatory properties. This study is the first to report green synthesis of ZnO/A. sativum nano-composite as an effective therapy in treating cryptosporidiosis which gave better results than using A. sativum alone. It provides an economical and environment-friendly approach towards novel delivery synthesis for antiparasitic applications. RESEARCH HIGHLIGHTS: Green synthesis of ZnO-NPs was developed using A. sativum extract. The morphology of prepared ZnO-NPs appeared as sponge with many pores on SEM The study evaluates its therapeutic efficacy against murine cryptosporidiosis The green synthesized ZnO-NPs significantly reduced percent of oocyst shedding, improved the pathological changes, and showed high antioxidant and anti-inflammatory potentials.

The effect of an injectable toltrazuril - gleptoferron (Forceris®) on Cystoisospora suis oocyst excretion and growth of neonatal piglets pre- and post-weaning.

In this study the efficacy of an intramuscular formulation of toltrazuril combined with gleptoferron for the control of porcine cystoisosporosis caused by Cystoisospora suis was investigated. The study was carried out on three Belgian farms with a confirmed history of C. suis infections. As none of the farms implemented a standardized toltrazuril treatment regimen for their piglets, the presence of resistant C. suis strains seems improbable. In total 90 litters, representing 1249 piglets, were included in the study and randomly allocated to either the treatment or control group. Piglets in the treatment group received a single intramuscular injection, containing 45 mg toltrazuril and 200 mg gleptoferron, between 1 and 3 days of age. Piglets in the control group received a single injection with only 200 mg gleptoferron. The effect of treatment on oocyst excretion, expressed in oocysts per gram of feces (OPG), average daily weight gain (ADG) and mortality was determined both pre- and post-weaning. A significant decrease in OPG as well as a decrease in the number of litters (pre-weaning) and pens (post-weaning) that tested positive for cystoisosporosis, was observed in the treated animals compared to the controls. Furthermore, treatment resulted in an increased ADG during the period from day 1 to day 21 (p-value: 0.03881). There was no significant difference in mortality observed between the treatment group to the control group (p-value: 0.2167). To our knowledge, this is the first report on the effect of toltrazuril on oocyst excretion after weaning. This finding highlights the potential long-term benefits of the treatment beyond the initial administration.

Aqueous Ozone Exposure Inhibits Sporulation in the Cyclospora cayetanensis Surrogate Eimeria acervulina.

Ozone is a potent disinfecting agent used to treat potable water and wastewater, effectively clearing protozoa such as Giardia and Cryptosporidium spp. It is unclear whether ozone treatment of water or fresh produce can reduce the spread of the emerging parasite Cyclospora cayetanensis, which causes cyclosporiasis in humans. Obtaining viable C. cayetanensis oocysts to evaluate inactivation methods is challenging because we lack the means to propagate them in vitro, because of delays in case reporting, and because health departments typically add inactivating fixatives to clinical specimens. Research in various surrogate organisms has sought to bolster understanding of the biology of C. cayetanensis. Among these surrogates is the poultry parasite Eimeria acervulina, a closely related and easily cultured parasite of economic significance. We used this surrogate to evaluate the consequences of ozone treatment, using the sporulation state as an indicator of infectious potential. Treating with ozonated water acidified with citric acid reduced sporulation ability in a dose-dependent manner; treatment with up to 4.93 mg/L initial concentration of ozone resulted in a 93% inactivation of sporulation by 7 days posttreatment. This developmental arrest was accompanied by transcriptional changes in genes involved in regulating the response to reactive oxygen species (ROS) in a time course that is consistent with the production of oxygen free radicals. This study shows that ozone is highly effective in preventing sporulation of E. acervulina, a model coccidian used as a surrogate for Cyclospora. Furthermore, ozone exposure induced molecular responses to general oxidative stress, documented with several well-characterized antioxidant enzymes.

Prophylactic and Therapeutic Efficacy of Ultrasonicated Rosmarinus officinalis Ethanolic Extract and its Chitosan-Loaded Nanoparticles Against Eimeria tenella Infected Broiler Chickens.

PURPOSE: The in vivo efficacy of ultrasonicated Rosmarinus officinalis ethanolic extract (UROEE) and its chitosan-loaded nanoparticles (UROEE-CsNPs) was investigated as a dietary prophylactic agent and as a therapeutic treatment against Eimeria tenella infected broiler chickens. METHODS: Chickens were infected with 4 × 104 E. tenella oocysts at 21 days old for primary infection and with 8 × 104 oocysts at 35 days old for secondary infection. Eleven experimental groups were conducted. Dietary addition of 100 mg/kg UROEE and 20 mg/kg for CsNPs as well as UROEE-CsNPs were included for prophylactic groups from day 1 to 42. The same doses were used for therapeutic treatment groups for 5 constitutive days. Oocyst output in feces was counted. Histopathological and immunohistochemical studies were conducted. Gene expression of pro-inflammatory cytokines as IFN-γ, IL-1ß and IL-6 as well as anti-inflammatory cytokines as IL-10 and TGF-ß4 was analyzed using semi-quantitative reverse transcriptase-PCR. RESULTS: The results showed an efficacy of UROEE, CsNPs and UROEE-CsNPs in reduction of oocyst excretion and improving the cecal tissue architecture. CD4+ and CD8+ T lymphocytes protein expression were reduced. E. tenella infection lead to upregulation of pro-inflammatory cytokines as IFN-γ, IL-1ß, IL-6 and anti-inflammatory cytokines as TGF-ß4 following primary infection, while their expression was downregulated following secondary infection. CONCLUSION: The dietary prophylactic additives and therapeutic treatments with UROEE, CsNPs and UROEE-CsNPs could decrease the inflammatory response to E. tenella as indicated by oocyst output reduction, histopathological improvements, CD4+ and CD8+ T cells protein expression reduction as well as reducing mRNA expression levels of the tested cytokines following primary and secondary infections. Consequently, these results will help to develop better-combating strategies for the control and prevention of coccidiosis on poultry farms as a dietary prophylactic agent or as a therapeutic treatment.

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

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

Assessment of the Activity of Decoquinate and Its Quinoline-O-Carbamate Derivatives against Toxoplasma gondii In Vitro and in Pregnant Mice Infected with T. gondii Oocysts.

The quinolone decoquinate (DCQ) is widely used in veterinary practice for the treatment of bacterial and parasitic infections, most notably, coccidiosis in poultry and in ruminants. We have investigated the effects of treatment of Toxoplasma gondii in infected human foreskin fibroblasts (HFF) with DCQ. This induced distinct alterations in the parasite mitochondrion within 24 h, which persisted even after long-term (500 nM, 52 days) treatment, although there was no parasiticidal effect. Based on the low half-maximal effective concentration (IC50) of 1.1 nM and the high selectivity index of >5000, the efficacy of oral treatment of pregnant mice experimentally infected with T. gondii oocysts with DCQ at 10 mg/kg/day for 5 days was assessed. However, the treatment had detrimental effects, induced higher neonatal mortality than T. gondii infection alone, and did not prevent vertical transmission. Thus, three quinoline-O-carbamate derivatives of DCQ, anticipated to have better physicochemical properties than DCQ, were assessed in vitro. One such compound, RMB060, displayed an exceedingly low IC50 of 0.07 nM, when applied concomitantly with the infection of host cells and had no impact on HFF viability at 10 µM. As was the case for DCQ, RMB060 treatment resulted in the alteration of the mitochondrial matrix and loss of cristae, but the changes became apparent at just 6 h after the commencement of treatment. After 48 h, RMB060 induced the expression of the bradyzoite antigen BAG1, but TEM did not reveal any other features reminiscent of bradyzoites. The exposure of infected cultures to 300 nM RMB060 for 52 days did not result in the complete killing of all tachyzoites, although mitochondria remained ultrastructurally damaged and there was a slower proliferation rate. The treatment of mice infected with T. gondii oocysts with RMB060 did reduce parasite burden in non-pregnant mice and dams, but vertical transmission to pups could not be prevented.

Effect of household bleach on the structure of the sporocyst wall of Toxoplasma gondii.

Toxoplasma gondii oocysts are responsible for food- and water-borne infections in humans worldwide. They are resistant to common chemical disinfectants, including chlorinated products, presumably due to the structure and molecular nature of the oocyst wall but also the sporocyst wall. In this study, we used fluorescence microscopy and transmission electron microscopy to characterise the structure of both the oocyst and sporocyst walls, exposed to household bleach. Bleach removed the outer layer of the oocyst wall and the outer layer of the wall of sporocysts exposed due to rupture of the oocyst wall. The loss of the outer sporocyst wall layer was associated with a decrease in its autofluorescence, which can be linked to the degradation of dityrosine cross-link proteins, and loss of Maclura pomifera lectin-reactive glycoproteins. This study suggests that the inner layers of the oocyst and sporocyst walls are the main structures responsible for the resistance of the parasite to household bleach.

TITLE: Effet de l'eau de Javel à usage domestique sur la structure de la paroi du sporocyste de Toxoplasma gondii. ABSTRACT: Les oocystes de Toxoplasma gondii sont responsables chez l'homme d'infections cosmopolites d'origine alimentaire et hydrique. Ils sont résistants aux désinfectants chimiques usuels, notamment aux produits chlorés, vraisemblablement en raison de la structure et de la nature moléculaire de la paroi de l'oocyste mais aussi de celle du sporocyste. Dans cette étude, nous avons utilisé la microscopie à fluorescence et la microscopie électronique à transmission pour caractériser la structure de la paroi des oocystes et des sporocystes exposés à l'eau de Javel à usage domestique. L'eau de Javel élimine la couche externe de la paroi de l'oocyste et la couche externe de la paroi des sporocystes exposés en raison de la rupture de la paroi de l'oocyste. La perte de la couche externe de la paroi du sporocyste est associée à une diminution de son autofluorescence, qui peut être liée à la dégradation de polymères protéiques de dityrosine, et à une perte des glycoprotéines réactives à la lectine Maclura pomifera. Cette étude suggère que les couches internes des parois de l'oocyste et du sporocyste sont les principales structures responsables de la résistance du parasite à l'eau de Javel à usage domestique.

Curcumin reduces enteric isoprostane 8-iso-PGF2α and prostaglandin GF2α in specific pathogen-free Leghorn chickens challenged with Eimeria maxima.

The purpose of this pilot study was to evaluate and determine the concentration of prostaglandin GF2α (PGF2α) and isoprostane 8-iso-PGF2α in plasma and intestine of specific pathogen-free (SPF) Leghorn chickens challenged with Eimeria maxima, with or without dietary supplementation of curcumin using solid-phase microextraction and ultra-performance liquid chromatography/tandem mass spectrometry. Eighty 1-day-old male SPF chickens were randomly allocated to one of four groups with four replicates (n = 5 chickens/replicate). Groups consisted of: (1) Control (no challenge), (2) Curcumin (no challenge), (3) Eimeria maxima (challenge), and (4) Eimeria maxima (challenge) + curcumin. At day 28 of age, all chickens in the challenge groups were orally gavaged with 40,000 sporulated E. maxima oocysts. No significant differences (P > 0.05) were observed in the groups regardless of the treatment or challenge with E. maxima. Enteric levels of both isoprostane 8-iso-PGF2α and PGF2α at 7 days and 9 days post-challenge were significantly increased (P < 0.01) compared to the non-challenge control chickens. Interestingly, the enteric levels of both isoprostane 8-iso-PGF2α and PGF2α at 7 days post-challenge were significantly reduced in chickens fed curcumin, compared to control chickens challenge with E. maxima. At 9 days post-challenge, only levels of isoprostane 8-iso-PGF2α in the enteric samples were significantly reduced in chickens challenged with E. maxima supplemented with curcumin, compared with E. maxima challenge chickens. No differences of isoprostane 8-iso-PGF2α or PGF2α were observed in plasma at both days of evaluation. Similarly, no significant differences were observed between the challenge control or chickens challenge with E. maxima and supplemented with curcumin at both times of evaluation. The results of this pilot study suggests that the antioxidant anti-inflammatory properties of curcumin reduced the oxidative damage and subsequent intestinal mucosal over-production of lipid oxidation products. Further studies to confirm and extend these results in broiler chickens are required.

Rapamycin inhibits pathogen transmission in mosquitoes by promoting immune activation.

Repeated blood meals provide essential nutrients for mosquito egg development and routes for pathogen transmission. The target of rapamycin, the TOR pathway, is essential for vitellogenesis. However, its influence on pathogen transmission remains to be elucidated. Here, we show that rapamycin, an inhibitor of the TOR pathway, effectively suppresses Plasmodium berghei infection in Anopheles stephensi. An. stephensi injected with rapamycin or feeding on rapamycin-treated mice showed increased resistance to P. berghei infection. Exposing An. stephensi to a rapamycin-coated surface not only decreased the numbers of both oocysts and sporozoites but also impaired mosquito survival and fecundity. Transcriptome analysis revealed that the inhibitory effect of rapamycin on parasite infection was through the enhanced activation of immune responses, especially the NF-κB transcription factor REL2, a regulator of the immune pathway and complement system. Knockdown of REL2 in rapamycin-treated mosquitoes abrogated the induction of the complement-like proteins TEP1 and SPCLIP1 and abolished rapamycin-mediated refractoriness to Plasmodium infection. Together, these findings demonstrate a key role of the TOR pathway in regulating mosquito immune responses, thereby influencing vector competence.

Effect of methyl gallate on immune response of Biomphalaria alexandrina (Ehrenberg, 1831) snails to infection with Schistosoma mansoni (Sambon, 1907).

Schistosomiasis still affects a lot of people in many developing countries. Reducing the disease dissemination has been the target of various studies. As methyl gallate has antioxidant properties, it is assumed that it can be a good candidate for stimulating the immune response of snails. So, the aim of this work is to investigate the potential of using methyl gallate as an immunostimulant to Biomphalaria alexandrina snails in order to prevent the development of invading miracidia into infective cercariae. The infected snails were exposed to three concentrations of methyl gallate for two periods: 24 and 72 h. The results indicated that the most effective concentration was the lowest one: 125 mg/L of methyl gallate for 72 h, as it reduced both infection rate and mean number of shed cercariae. Also, it increased the total number of snails' hemocytes in hemolymph, which were observed in head-foot region and digestive gland of treated snails surrounding degenerated sporocysts and cercariae. In addition, hydrogen peroxide showed its highest content in tissues of snails exposed to 125 mg/L of methyl gallate for 72 h. In conclusion, methyl gallate can be considered as one of the most promising immunostimulants of B. alexandrina snails against infection with Schistosoma mansoni.

Inactivation of Cryptosporidium parvum oocysts and faecal indicator bacteria in cattle slurry by addition of ammonia.

AIMS: To determine inactivation of Cryptosporidium parvum oocysts and reduction of Escherichia coli and enterococci in cattle slurry added aqueous ammonia. METHODS AND RESULTS: Escherichia coli, enterococci and nonviable C. parvum oocysts (DAPI+PI+) were enumerated every second day for 2 weeks in cattle slurry amended with 60 mmol l-1 aq. ammonia and compared with untreated slurry at three temperatures. Regardless of temperature, the proportion of nonviable C. parvum oocysts increased significantly faster over time in slurry with added ammonia than raw slurry (P = 0·021) corresponding to 62·0% higher inactivation (P = 0·001) at day 14. Additionally, 91·8% fewer E. coli and 27·3% fewer enterococci were observed in slurry added ammonia at day 14 compared to raw slurry. CONCLUSION: The addition of aqueous ammonia to raw slurry significantly reduced the viability of C. parvum oocysts and numbers of bacterial indicators. Hence, ammonia is usable at lower pathogen concentrations in slurry before application to agricultural land. SIGNIFICANCE AND IMPACT OF THE STUDY: Livestock waste is a valuable source of plant nutrients and organic matter, but may contain high concentrations of pathogens like E. coli and Cryptosporidium sp. that can be spread in the environment, and cause disease outbreaks. However, die-off rates of pathogens in organic waste can increase following increasing ammonia concentrations.

Effects of probiotic and Bidens pilosa on the performance and gut health of chicken during induced Eimeria tenella infection.

AIM: In this study, we have examined the individual and combined protective mechanism of probiotic and Bidens pilosa on the performance and gut health of chickens during Eimeria tenella infection over a 29-day experimental trial. METHODS AND RESULTS: A total of one hundred and fifty 1-day-old chickens were equally distributed into five treatment groups with three biological replicates: two groups were allocated as control groups (control group untreated unchallenged, CG and control positive untreated challenged, CPG) and three groups were fed diets with probiotic (PG), B. pilosa (BPG) and probiotic + B. pilosa (PG + BPG) and challenged with E. tenella. Birds of all groups were assessed for pre and post-infection body weights, oocysts shedding, caecal lesion scores and mRNA expression levels of apoptosis related proteins (Bcl-2, Bax and caspase-3), antioxidant enzymes (CAT and SOD 1), pro-inflammatory cytokines (IL-6 and IL-8) and tight junction proteins (CLDN 1 and ZO 1). Our results revealed that during infection (day 21-29), E. tenella challenged chickens significantly decreased the body weight compared with uninfected control chickens; however, there was no significant effect on body weight of chickens fed with probiotic, B. pilosa and probiotic + B. pilosa was observed. Eimeria tenella challenged untreated birds increased (P < 0·05) oocysts shedding, destructive ratio of caeca and mortality as compared to treated challenged birds. CPG group up-regulated the mRNA expression levels of anti-apoptosis protein Bcl-2 while down-regulated the pro-apoptosis protein Bax relative to PG, BPG and PG + BPG groups. Moreover chickens fed probiotic, B. pilosa and probiotic + B. pilosa diets enhanced the activities of antioxidant enzymes, pro-inflammatory cytokines and tight junction proteins with the comparison of control positive untreated challenged chickens. CONCLUSION: These findings elaborated that feed supplementation of probiotic and B. pilosa (individually or in combination) appeared to be effective in inhibiting the occurrence of disease and decreasing the severity of Eimeria infection in chickens. SIGNIFICANCE AND IMPACT OF THE STUDY: This study explained the underlying anti-coccidial mechanism in which probiotic and B. pilosa (individually and/or in combination) improve the performance of chicken and protect against gut inflammatory responses caused by E. tenella.

Heparin Administered to Anopheles in Membrane Feeding Assays Blocks Plasmodium Development in the Mosquito.

Innovative antimalarial strategies are urgently needed given the alarming evolution of resistance to every single drug developed against Plasmodium parasites. The sulfated glycosaminoglycan heparin has been delivered in membrane feeding assays together with Plasmodium berghei-infected blood to Anopheles stephensi mosquitoes. The transition between ookinete and oocyst pathogen stages in the mosquito has been studied in vivo through oocyst counting in dissected insect midguts, whereas ookinete interactions with heparin have been followed ex vivo by flow cytometry. Heparin interferes with the parasite's ookinete-oocyst transition by binding ookinetes, but it does not affect fertilization. Hypersulfated heparin is a more efficient blocker of ookinete development than native heparin, significantly reducing the number of oocysts per midgut when offered to mosquitoes at 5 µg/mL in membrane feeding assays. Direct delivery of heparin to mosquitoes might represent a new antimalarial strategy of rapid implementation, since it would not require clinical trials for its immediate deployment.

Anticoccidial effect of Fructus Meliae toosendan extract against Eimeria tenella.

CONTEXT: Fructus Meliae toosendan extracts (FMTE) have a good therapeutic effect on coccidiosis, but there is no relevant research on its prophylactic effect on coccidiosis. OBJECTIVE: This study comprehensively evaluates the anticoccidial effect of FMTE. MATERIALS AND METHODS: In vitro, the unsporulated oocysts were treated with serial dilutions of FMTE and incubated for 7 d, and the sporulated oocysts were counted for calculating the median lethal concentration (LC50) of FMTE. In vivo, 180 10-day-old broiler chickens free of coccidiosis were weighted and randomly distributed into six groups: normal group, untreated group, 4 protective groups (positive group and three FMTE groups). From day 10 to day 21, chickens in the three FMTE groups were pre-treated with FMTE at the dosage of 2.5, 5 and 10 g/kg/d, respectively, and chickens in the positive group were pre-treated with qiuliling (10 g/kg/d). On day 14, chickens in all groups except the normal group were orally infected with 1.5 × 104 sporulated oocysts. The clinical symptoms were observed from day 10 to day 21, the anticoccidial index (ACI), tissue lesions, and intestinal microflora were determined on day 21. RESULTS: FMTE showed anti-sporulation effect against E. tenella and the LC50 value was 245.83 µg/mL in vitro. In vivo, FMTE at the dosage of 10 g/kg/d was effective against E. tenella infection, and its ACI value was 162.56, which was higher than the value of positive drug qiuliling (128.81). Discussion and conclusions: FMTE have potent anticoccidial effects, and it presents an alternative anticoccidial agent for avian coccidiosis control.

Efficacy of dietary supplemental insoluble fibrous materials in ameliorating adverse effects of coccidial challenge in broiler chickens.

The current experiment was designed to examine effects of dietary supplemental sunflower hulls (SH) and rice hulls (RH) on growth performance, carcass traits, intestinal morphology, lesion score and oocyst shedding in broiler chickens exposed to coccidial challenge. A total of 540 broiler chickens (Ross 308) were assigned to six dietary treatments based on a factorial arrangement (2 × 3) across 1-14, 14-28 and 28-42-d periods. Experimental treatments consisted of broiler chickens without or with coccidial challenge each offered with three different diets: a basal diet or basal diet supplemented with either RH or SH at 40 g/kg diet, respectively. Infection with Eimeria impaired daily weight gain (DWG) and feed conversion ratio (FCR) of broiler chickens during growing period (p < 0.05) while supplementation of SH or RH reduced the adverse effect of coccidiosis so that birds had similar DWG to those fed the basal diet without infection. However, only dietary SH improved the FCR of broilers challenged with coccidiosis. Regardless of coccidial challenge, dietary access to insoluble fibre improved performance of broilers across the growing period (p < 0.05); however, this effect was not observed during the entire rearing period. Relative weights of liver and pancreas were increased in birds subjected to coccidial challenge on d 21 of age (p < 0.05). Moreover, relative weights of the intestinal segments were enhanced (p < 0.05). Furthermore, gizzard weights were higher in birds receiving diets added with fibre (p < 0.05). Infection with coccidiosis decreased villus height and villus height to crypt depth ratio in duodenum of broilers which received the basal diet compared with those fed the same feed without coccidial challenge (p < 0.05). However, supplemental SH could decrease the negative effect of infection on the noted intestinal morphometric attributes. Similarly, a marked reduction was observed for lesion score and faecal oocyst excretion of challenged broilers fed on dietary supplemental fibre (p < 0.05). In conclusion, supplementation of insoluble fibre could ameliorate negative effects of coccidial challenge on DWG of broiler chickens and inclusion of SH in diet of birds exposed to Eimeria infection could be recommended.

Efficacy of probiotic Enterococcus faecium in combination with diclazuril against coccidiosis in experimentally infected broilers.

AIMS: The study was conducted to investigate the combination of a probiotic strain of Enterococcus faecium and diclazuril to control coccidiosis in broilers. METHODS AND RESULTS: A total of 240 one-day-old female broiler chicks were divided into eight groups (30 chicks per group): prophylactic groups (G1, G2 and G3) and therapeutic groups (G4, G5 and G6) and two control groups (untreated infected, G7 and untreated uninfected, G8 controls). In the prophylactic approach, diclazuril alone (G1), probiotic alone (G2) or a mixture of both probiotic and diclazuril (G3) was orally administered to the chicks via drinking water 10 days prior to the infection. However, in the therapeutic approach, G4, G5 and G6 birds were administered diclazuril alone, probiotic alone and diclazuril+probiotic mix, respectively, in drinking water for five consecutive days after the appearance of clinical signs of coccidiosis. Birds of both approaches and G7 were experimentally infected with 25 × 103 Eimeria-sporulated oocysts. Chicks in G3 showed the highest weight gain, the lowest lesion score, a low oocyst count and mortality rate among the challenged groups. Moderate lesion scores and oocyst counts were observed in chickens administered probiotics prophylactically. In the therapeutic approach, broilers in G6 but not G5 displayed a decreased mortality rate and lesion score in comparison to those in G7 and G8. However, the result of the probiotic-treated group was not significantly different from that in the untreated infected control group. CONCLUSION: The probiotic supplementation as a prophylactic approach can decrease the adverse effects of eimerian infection. In addition, the probiotic and diclazuril mix achieved a considerable improvement in the growth performance. Therefore, probiotic plus diclazuril combination achieved a synergistic effect. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigation into the synergism/antagonism between a probiotic and diclazuril as anticoccidial agent and the difference in the timing of administration.

The anticoccidial activity of the fluoroquinolone lomefloxacin against experimental Eimeria tenella infection in broiler chickens.

Coccidiosis is a crucial parasitic disease of the poultry industry. As a result of the enormous global economic losses and the increased resistance to the conventional anticoccidial agents, there is a continuous need to find new anticoccidials. Here, the anticoccidial effect of the fluoroquinolone lomefloxacin versus diclazuril in experimentally infected broilers was tested for the treatment of Eimeria tenella infection. Ninety 14-day-old Cobb strain broiler chickens were allocated into five groups, each with 18 chicks. Group 1 (G1) was separated as an uninfected negative control and received no treatment; group 2 (G2), infected untreated (positive control); group 3 (G3), infected and treated with lomefloxacin at a dose rate of 100 ppm in drinking water; group 4 (G4), infected and treated with diclazuril at a dose rate of 2.5 ppm in drinking water; group 5 (G5), infected and treated with lomefloxacin at a dose rate of 100 ppm plus diclazuril at dose rate of 2.5 ppm in drinking water. Clinical signs, mortality rates, number of oocysts per gram of faeces (OPG), growth performance parameters (weight gain: WG and feed conversion ratio: FCR), lesion scoring, haematological and serum biochemical analyses, antioxidant biomarkers and histopathologic inspection of the caeca were used as evaluation criteria for the anticoccidial efficacy of both lomefloxacin and diclazuril. The findings herein showed that administration of lomefloxacin and/or diclazuril improved growth performance parameters (WG, FCR) and significantly (P ≤ 0.05) reduced OPG, and diminished the severity of bloody diarrhoea and mortalities. Additionally, haematological indices and serum biochemical parameters such as ALT, AST, ALP, creatinine, uric acid, total proteins, albumin and globulin were improved. Finally, a significant elevation in the levels of the antioxidant biomarkers was observed in the chicks of G3, G4 and G5 as compared with those of G2.

Effect of diet supplemented with coconut essential oil on performance and villus histomorphology in broiler exposed to avian coccidiosis.

The current research study was designed to determine the inclusion of 2% dietary essential coconut oil with and without coccidiosis challenge on performance, carcass characteristics, and intestinal histomorphology in broilers. A total of 560 broiler chicks were divided into 4 groups and then subdivided into 5 replicates. Coconut oil was used at 2% in feed, whereas coccidiosis challenged was introduced using 30,000 oocysts. The other four groups were designated as G1 (without coconut oil and without oocysts), G2 (without coconut oil with oocysts), G3 (with coconut oil without oocysts), and G4 (with coconut oil and with oocysts). The results revealed that the overall feed consumption was significantly (P < 0.01) increased in G1 and G2 than G3 and G4 groups. Overall weight gain was significantly (P < 0.01) higher in G3 compared with all other groups. Significantly (P < 0.01) better feed conversion ratio was recorded at the finisher phase in G3 and G4 groups in comparison with G1 and G2. The villus length, width, and surface area were higher (P < 0.01) in G3 compared with G2. Based on the findings of the present study, it was concluded that the use of 2% coconut oil in broiler feed improved growth performance and villus histology during coccidial challenge.

Calcium-dependent Protein Kinases in Malaria Parasite Development and Infection.

Apicomplexan parasites have challenged researchers for nearly a century. A major challenge to developing efficient treatments and vaccines is the parasite's ability to change its cellular and molecular makeup to develop intracellular and extracellular niches in its hosts. Ca2+ signaling is an important messenger for the egress of the malaria parasite from the infected erythrocyte, gametogenesis, ookinete motility in the mosquito, and sporozoite invasion of mammalian hepatocytes. Calcium-dependent protein kinases (CDPKs) have crucial functions in calcium signaling at various stages of the parasite's life cycle; this therefore makes them attractive drug targets against malaria. Here, we summarize the functions of the various CDPK isoforms in relation to the malaria life cycle by emphasizing the molecular mechanism of developmental progression within host tissues. We also discuss the current development of anti-malarial drugs, such as how specific bumped kinase inhibitors (BKIs) for parasite CDPKs have been shown to reduce infection in Toxoplasma gondii, Cryptosporidium parvum, and Plasmodium falciparum. Our suggested combinations of BKIs, artemisinin derivatives with peroxide bridge, and inhibitors on the Ca(2+)-ATPase PfATP6 as a potential target should be inspected further as a treatment against malaria.