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

Ticks pose a significant threat due to their ability to lay thousands of eggs, which can persist in the environment for extended periods. While the impact of various compounds on adult and larval ticks has been studied, research on their efficacy against tick eggs is limited. This study evaluated the ovicidal activity of commercial acaricides, disinfectants, and natural products against Rhipicephalus annulatus eggs using the egg hatch assay (EHA). Deltamethrin and cypermethrin caused a non-significant inhibition of hatching (IH%), even at concentrations higher than the recommended levels. By contrast, the acaricides chlorpyrifos, phoxim, and amitraz significantly inhibited hatching at all tested concentrations. Ivermectin also demonstrated significant IH% at various concentrations but did not fully inhibit the hatching process. Among the disinfectants tested, Virkon-S®, TH4, and Chlorox showed insignificant effects, whereas formalin achieved an IH% of only 34.1% at a high concentration of 200 mg/mL. Natural products, carvacrol and thymol, exhibited significant ovicidal activity, with a significant IH%. In a semi-field application, phoxim (0.5 mg/mL) and deltamethrin (0.05 mg/mL) were sprayed on tick eggs on pasture soil from a farm. The results indicated that phoxim-treated eggs had a 40% IH%, while deltamethrin-treated eggs showed only an 8.79% IH%. In conclusion, the acaricides amitraz, phoxim, and chlorpyrifos, as well as the natural products carvacrol and thymol, caused significant toxicity to R. annulatus eggs.

Tribolium castaneum is a challenging pest of stored products, causing significant economic losses. The present study explored the efficacy of Coridothymus capitatus essential oil and its primary constituent, carvacrol, as eco-friendly alternatives for managing this pest. To evaluate their insecticidal potential, repellency, fumigant toxicity, and antifeedant properties, progeny inhibition assays were performed. Carvacrol exhibited superior repellency compared to the essential oil, achieving a 92% repellency rate at 2 mg/cm2. Both compounds demonstrated significant fumigant toxicity against T. castaneum, with LC50 values of 168.47 and 106.5 μL/L for the essential oil and carvacrol, respectively, after 24 h. Carvacrol also outperformed the essential oil in antifeedant activity, inducing an 80.7% feeding deterrence at 1.17 mg/g. Moreover, both treatments effectively suppressed the development of the pest’s progeny. These results collectively underscore the potent insecticidal properties of C. capitatus essential oil and carvacrol, particularly carvacrol, as promising candidates for the sustainable management of T. castaneum in stored product protection.

IntroductionRecently, the use of botanicals as an alternative to coccidiostats has been an appealing approach for controlling coccidiosis. Therefore, this study was conducted to evaluate the potential role of aqueous methanolic extract (200 mg/kg) of Krameria lappacea (roots) (KLRE) against infection induced by Eimeria papillata.MethodsA total of 25 male C57BL/6 mice were divided into five groups (I, II, III, IV, and V). On 1st day of the experiment, all groups except groups I (control) and II (non-infected-treated group with KLRE), were inoculated orally with 103 sporulated E. papillata oocysts. On the day of infection, group IV was treated with KLRE. Group V served as an infected-treated group and was treated with amprolium (coccidiostat).ResultsTreatment with extract and coccidiostat was continued for five consecutive days. While not reaching the efficacy level of the reference drug (amprolium), KLRE exhibited notable anticoccidial activity as assessed by key criteria, including oocyst suppression rate, total parasitic stages, and maintenance of nutrient homeostasis. The presence of phenolic and flavonoid compounds in KLRE is thought to be responsible for its positive effects. The Eimeria infection increased the oxidative damage in the jejunum. KLRE treatment significantly increased the activity of catalase and superoxide dismutase. On the contrary, KLRE decreased the level of malondialdehyde and nitric oxide. Moreover, KLRE treatment decreased macrophage infiltration in the mice jejunal tissue, as well as the extent of CD4 T cells and NFkB. E. papillata caused a state of systemic inflammatory response as revealed by the upregulation of inducible nitric oxide synthase (iNOs)-mRNA. Upon treatment with KLRE, the activity of iNOs was reduced from 3.63 to 1.46 fold. Moreover, KLRE was able to downregulate the expression of pro-inflammatory cytokines interferon-γ, nuclear factor kappa B, and interleukin-10 -mRNA by 1.63, 1.64, and 1.38 fold, respectively. Moreover, KLRE showed a significant reduction in the expression of IL-10 protein level from 104.27 ± 8.41 pg/ml to 62.18 ± 3.63 pg/ml.ConclusionCollectively, K. lappacea is a promising herbal medicine that could ameliorate the oxidative stress and inflammation of jejunum, induced by E. papillata infection in mice.

Abstract Coccidiosis is the most prevalent disease-causing widespread economic loss among farm and domestic animals. Currently, several drugs are available for the control of this disease but resistance has been confirmed for all of them. There is an urgent need, therefore, for the identification of new sources as alternative treatments to control coccidiosis. The present work aimed to study the effect of the Persea americana extract (PAE) as an anti-coccidial, anti-oxidant, and anti-apoptotic modulator during murine intestinal Eimeria papillata infection. A total of 25 male mice were divided into five groups, as follows: Group1: Non-infected-non-treated (negative control), Group2: Non-infected-treated group with PAE (500 mg/kg b.w). Group3: Infected-non-treated (positive control), Group4: Infected-treated group with PAE (500 mg/kg b.w.), and Group5: Infected-treated group with Amprolium (120 mg/kg b.w.). Groups (3–5) were orally inoculated with 1 × 103 sporulated E. papillata oocysts. After 60 min of infection, groups (4 and 5) were treated for 5 consecutive days with the recommended doses of PAE and amprolium. The fact that PAE has an anti-coccidial efficacy against intestinal E. papillata infection in mice has been clarified by the reduction of fecal oocyst output on the 5th day post-infection by about 85.41%. Moreover, there is a significant reduction in the size of each parasite stage in the jejunal tissues of the infected-treated group with PAE. PAE counteracted the E. papillata-induced loss of glutathione peroxidase (GPx), superoxide dismutase (SOD), and total antioxidant capacity (TCA). E. papillata infection also induced an increase in the apoptotic cells expressed by caspase-3 which modulated after PAE treatment. Moreover, the mRNA expression of the goblet cell response gene, mucin (MUC2), was upregulated from 0.50 to 1.20-fold after treatment with PAE. Based on our results, PAE is a promising medicinal plant with anti-coccidial, anti-oxidant, and anti-apoptotic activities and could be used as a food additive.

Eimeria spp. are responsible for the economic loss of both domestic and wild animals due to coccidiosis, the most common parasitic disease. The resistance to currently available drugs used to treat coccidiosis has been proven. Medicinal plants that contain physiologically active phytochemicals have been widely used in traditional medicine. Teucrium polium leaf extract (TPLE) has been shown to exhibit pharmacological, antioxidant, and anticoccidial properties in different experiments. Here, our investigation focused on how T. polium leaf extract affected the way that Eimeria papillate caused intestinal injury in mice. Thirty-five male Swiss albino mice were divided into seven groups, as follows: group I: untreated and uninfected (negative control); group II: uninfected, treated group with TPLE (150 mg/kg b.w); and group III: infected untreated (positive control). Groups III–VII were orally administered 103 sporulated E. papillata oocysts. A total of 60 min after infection, groups IV–VI were treated for five successive days with 50, 150, and 250 mg/kg b.w TPLE, respectively, while group VII was treated with amprolium (120 mg/kg b.w.). The mice had been euthanized on the fifth day post-infection, and the jejunum tissues were prepared for histology and oxidative stress studies. A total of 150 mg/kg of TPLE was the most effective dosage, significantly decreasing oocyst output by about 80.5%, accompanied by a significant reduction in the number of developmental parasitic phases in jejunal sections. In addition, the decrease in the number of goblet cells in the jejuna of mice raised after treatment. Also, TPLE greatly diminished the body weight loss of infected mice. Moreover, our research proved that TPLE reduced oxidative damage due to E. papillata infection via decreasing intestinal malondialdehyde (MDA) and nitric oxide (NO) levels and increasing reduced superoxide dismutase (SOD) and glutathione (GSH) levels. These results demonstrated that TPLE had potent anticoccidial properties. TPE’s efficacy as a natural antioxidant has also been demonstrated in reducing oxidative stress and enhancing antioxidant systems to mitigate biochemical and histological changes in the jejunum caused by E. papillata.

Eimeria is among the most harmful parasites that affect domestic animals. The development resistance in Eimeria to the majority of regularly used anticoccidials necessitates development of an effective alternative strategy. This study was therefore aimed to investigate the therapeutic efficacy of essential oils combination, Pelargonium graveolens (geranium) with thymol (PGT) against Eimeria papillata murine model. Three doses of 50, 100, and 150 mg/kg were tested in Eimeria experimentally infected mice to determine the combination's highest effective dose. The results showed that the 150 mg/kg dose was the most effective, reducing the oocyst output by 56.92 % (P ≤ 0.001). This result has been associated with a 66.19 % (P ≤ 0.001) reduction in the jejunal developmental stages. The antioxidant effect of PGT decreased lipid peroxidation (P ≤ 0.05) and increasing reduced glutathione (GSH) (P ≤ 0.001). Moreover, PGT decreased the jejunum's apoptotic cell count (P ≤ 0.001). The in vitro results showed that PGT significantly inhibited oocysts sporulation and destructed 96.33 % of treated oocysts. In conclusion, PGT has therapeutic effect as an anticoccidial alternative to control murine eimeriosis.

Abstract Background Recently, an increasing number of ichthyophthiriasis outbreaks has been reported, leading to high economic losses in fisheries and aquaculture. Although several strategies, including chemotherapeutics and immunoprophylaxis, have been implemented to control the parasite, no effective method is available. Hence, it is crucial to discover novel drug targets and vaccine candidates against Ichthyophthirius multifiliis. For this reason, understanding the parasite stage biology, host–pathogen interactions, molecular factors, regulation of major aspects during the invasion, and signaling pathways of the parasite can promote further prospects for disease management. Unfortunately, functional studies have been hampered in this ciliate due to the lack of robust methods for efficient nucleic acid delivery and genetic manipulation. In the current study, we used antisense technology to investigate the effects of targeted gene knockdown on the development and infectivity of I. multifiliis. Antisense oligonucleotides (ASOs) and their gold nanoconjugates were used to silence the heat shock protein 90 (hsp90) of I. multifiliis. Parasite stages were monitored for motility and development. In addition, the ability of the treated parasites to infect fish and cause disease was evaluated. Results We demonstrated that ASOs were rapidly internalized by I. multifiliis and distributed diffusely throughout the cytosol. Knocking down of I. multifiliis hsp90 dramatically limited the growth and development of the parasite. In vivo exposure of common carp (Cyprinus carpio) showed reduced infectivity of ASO-treated theronts compared with the control group. No mortalities were recorded in the fish groups exposed to theronts pre-treated with ASOs compared with the 100% mortality observed in the non-treated control fish. Conclusion This study presents a gene regulation approach for investigating gene function in I. multifiliis in vitro. In addition, we provide genetic evidence for the crucial role of hsp90 in the growth and development of the parasite, suggesting hsp90 as a novel therapeutic target for successful disease management. Further, this study introduces a useful tool and provides a significant contribution to the assessing and understanding of gene function in I. multifiliis.

The present study investigated the effect of environmental variables on the abundance and distribution of zooplankton in different habitats along the Nile River in Shattura Village. Zooplankton samples were collected from three distinct sites along the Nile River in Shattura Village, each exhibiting different environmental characteristics: Site 1 with vegetation, site 2 lacking vegetation, and site 3 being a drain canal. The study spanned from spring 2020 to winter 2021, during which the physico-chemical parameters of the water were analyzed. Rotifera constituted the majority of the zooplankton (54.73%), followed by Cladocera (20.59%), Copepoda (13.1%), and Ostracoda (8.9%). Among the 52 identified zooplankton species, Rotifera comprised 18 species, Cladocera 13 species, Copepoda 10 species, and Ostracoda 11 species. Site 1 exhibited the highest zooplankton density (44.08%), attributed to the presence of vegetation, followed by site 3 (37.18%), influenced by agricultural drains, and site 2 had the lowest density (18.73%). Zooplankton abundance peaked in summer and declined in winter. Notably, Rotifera abundance increased in populated sites (site 3), whereas other zooplankton groups thrived in less populated areas. Correlation analyses revealed positive associations between Rotifera and Cladocera with electric conductivity, total dissolved solids (TDS), chloride (Cl), calcium (Ca), and sulfate (SO4). Conversely, Copepoda showed positive correlations with water transparency, pH, dissolved oxygen, and biological oxygen demand (BOD). Overall, seasonal variations significantly impacted the zooplankton community, with Rotifera dominating and Ostracoda being the least abundant. High values of the Shannon–Weaver diversity index, richness, and evenness suggested ample food resources and favorable growth conditions. Transparency, conductivity, pH, dissolved oxygen, and BOD were identified as the key influencing parameters on zooplankton abundance. Additionally, vegetation and agricultural drains strongly influenced total zooplankton levels.

The epidemiology of parasite infection in local and imported breeds is quite an essential topic in the meat industry and human health. This study aims to determine the prevalence of Dicrocoelium dendriticum in local sheep breeds (Naemi, Najdi, and Harri) and imported breeds from Romania (Romani breed) and the epidemiology of the infection in Saudi Arabia. Morphological description, the relationship between dicrocoeliasis and sex, age, and histological changes were also presented. A total of 6845 slaughtered sheep at Riyadh Automated slaughterhouse were investigated and followed up for 4 months between 2020–2021. It included 4,680 local breeds and 2,165 imported Romanian breeds. Fecal samples and livers and gallbladders from slaughtered animals were examined for apparent pathological lesions. The results indicated that the infection rate in slaughtered animals was 10.6% in imported Romani sheep and 0.9% in the local Naeimi breed. After identifying the parasite morphologically, negative results were obtained from examining feces, gallbladders, and livers of Najdi and Harry sheep breeds. The mean number of eggs per 20 µL/gallbladder was low (72.78 ± 17.8: 76.11 ± 5.07), medium (334.59 ± 90.6: 292.91 ± 26.63), and high (1113.2 ± 22.3: 1004 ± 143.4) in imported and Naeime sheep, respectively. Significant differences were found between gender and age (males and females were 3.67% and 6.31%; > 2 years 4.39%, 1–2 years 4.22%, and 1 year 3.53%) respectively. Histopathological lesions in the liver were more pronounced. Our survey confirmed the presence of D. dendriticum in imported Romani and local Naeimi sheep, and the potential role of imported sheep in the epidemiology of dicrocoeliasis in Saudi Arabia.

Abstract The aim of the present study was to evaluate in vitro and in vivo the acaricidal activity of two forms of geranium (Pelargonium graveolens) (PG). These two forms were the P. graveolens essential oil nanoemulsion (PGN), and the PG in combination with the sesame oil (SO), PGSO). These forms were first evaluated in vitro for their adulticidal, ovicidal, and larvicidal activities against the different stages of acaricide-resistant Rhipicephalus annulatus (Say). Geranium nanoemulsion was prepared and then characterized by UV–Vis spectrophotometer, and zeta droplet size measurement. The results revealed that LC50 of the PG against the adult ticks was attained at concentration of 7.53% while it was decreased to 1.91% and 5.60% for PGSO and PGN, respectively. Also, the LC50 of PGN and PGSO were reached at concentrations of 1.688 and 0.944%, respectively against the larvae while the LC50 of the PG was reached at concentration of 3.435% for. The combination of PGN with PG exhibited non-significant ovicidal effect meanwhile PGSO showed significant ovicidal effect even at the low concentration (2.5%). The PGSO and PGN formulations were applied in a field trial to control the ticks of the naturally infested cattle. PGSO and PGN significantly reduced the tick burden to 74.83% and 87.97%, respectively at 3 weeks post-application with performance better than the deltamethrin (29.88%). In conclusion, the two PG forms can be used as suitable alternatives to control R. annulatus tick and they need further modifications for effective field application.

IntroductionTraditional approaches to collecting large-scale biodiversity data pose huge logistical and technical challenges. We aimed to assess how a comparatively simple method based on sequencing environmental DNA (eDNA) characterises global variation in plant diversity and community composition compared with data derived from traditional plant inventory methods.MethodsWe sequenced a short fragment (P6 loop) of the chloroplast trnL intron from from 325 globally distributed soil samples and compared estimates of diversity and composition with those derived from traditional sources based on empirical (GBIF) or extrapolated plant distribution and diversity data.ResultsLarge-scale plant diversity and community composition patterns revealed by sequencing eDNA were broadly in accordance with those derived from traditional sources. The success of the eDNA taxonomy assignment, and the overlap of taxon lists between eDNA and GBIF, was greatest at moderate to high latitudes of the northern hemisphere. On average, around half (mean: 51.5% SD 17.6) of local GBIF records were represented in eDNA databases at the species level, depending on the geographic region.DiscussioneDNA trnL gene sequencing data accurately represent global patterns in plant diversity and composition and thus can provide a basis for large-scale vegetation studies. Important experimental considerations for plant eDNA studies include using a sampling volume and design to maximise the number of taxa detected and optimising the sequencing depth. However, increasing the coverage of reference sequence databases would yield the most significant improvements in the accuracy of taxonomic assignments made using the P6 loop of the trnL region.

Tribolium castaneum is a damaging pest of stored grains, causing significant losses and secreting lethal quinones, which render the grains unfit for human consumption. Chemical insecticides are the most commonly used approach for control; however, they create insecticide resistance and affect the health of humans, animals, and the environment. As a result, it is critical to find an environmentally friendly pest-management strategy. In this study, two naturally occurring chemicals, benzyl alcohol (BA) and benzoyl benzoate (BB), were investigated for insecticidal activity against T. castaneum using different assays (impregnated-paper, contact toxicity, fumigant, and repellency assays). The results showed that BA had a significant insecticidal effect, with the LC50 achieved at a lower concentration in the direct-contact toxicity test (1.77%) than in the impregnated-paper assay (2.63%). BB showed significant effects in the direct-contact toxicity test, with an LC50 of 3.114%, and a lower toxicity in the impregnated-paper assay, with an LC50 of 11.75%. Furthermore, BA exhibited significant fumigant toxicity against T. castaneum, with an LC50 of 6.72 µL/L, whereas BB exhibited modest fumigant toxicity, with an LC50 of 464 µL/L. Additionally, at different concentrations (0.18, 0.09, 0.045, and 0.0225 µL/cm2), BA and BB both showed a notable and potent repelling effect. BA and BB significantly inhibited acetylcholinesterase, reduced glutathione (GSH), and increased malondialdehyde (MDA) in treated T. castaneum. This is the first report of BA insecticidal activity against the red flour beetle. Also, the outcomes of various assays demonstrated that the application of BA induces a potent bio-insecticidal effect. BA may be a promising eco-friendly alternative to control T. castaneum due to its safety and authorization by the EFSA (European Food Safety Authority).

This study investigated the anticoccidial activity of spinach (Spinacia oleracea) whole-plant extract against Eimeria tenella, both in vitro and in vivo. For this purpose, one hundred 8-day-old broiler chicks of both sexes were divided into four groups (n = 25 in each group). Chicks in the first group served as the negative control (non-treated–non-infected). Chicks in the second group were challenged at 18 days old with 5 × 104 E. tenella sporulated oocysts. The third group was challenged with 5 × 104 sporulated E. tenella oocysts at 18 days old after receiving spinach extract at a dose of 50 mg/kg at 8 days old. The fourth group received 0.2 mg/kg diclazuril (Coxiril® 0.2%) in their diet two days before being orally infected with 5 × 104 sporulated E. tenella oocysts and this continued till day 10 post-infection (PI). The growth performance, clinical symptoms, oocyst shedding, histological findings, and biochemical parameters were used to evaluate the efficacy on day 8 PI when the infection was at its peak. A gas chromatography examination revealed that omega-3 fatty acids were the main constituents of the spinach extract, followed by oleic acid, palmitic acid, and phytol, with amounts of 23.37%, 17.53%, 11.26%, and 7.97%, respectively. The in vitro investigation revealed that the spinach extract at concentrations of 10% and 5% inhibited the oocyst sporulation by 52.1% and 45.1%, respectively. The 5% concentration was selected for the in vivo trial based on the results of the in vitro study. The infected–untreated group showed high levels of OPG; lower body weight; a greater number of parasite stages; few goblet cells; decreased SOD, CAT, and GPX levels; and increased MDA and NO levels. The spinach-treated group, on the other hand, showed a significant decrease in oocyst output per gram of feces (OPG), increased body weight, decreased parasitic stages, and a nearly normal number of goblet cells. Additionally, it reduced malondialdehyde (MDA) and nitric oxide (NO), while increasing superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX). In conclusion, spinach produced significant antioxidant effects, increased body weight, reduced the number of oocysts and parasite stages in the caecum, and restored the number of goblet cells relative to those of an uninfected control. Furthermore, spinach extract inhibits the sporulation percentage of E. tenella oocysts. The ethanolic extract of S. oleracea (whole plant) contained high concentrations of fatty acids, palmitic acid, Phytol, betulin, and ursolic aldehyde, all of which are known to regulate the antioxidant pathway and modulate inflammatory processes and may be the main reason for its anticoccidial activity.

One of the most crucial approaches for treating human diseases, particularly parasite infections, is nanomedicine. One of the most significant protozoan diseases that impact farm and domestic animals is coccidiosis. While, amprolium is one of the traditional anticoccidial medication, the advent of drug-resistant strains of Eimeria necessitates the development of novel treatments. The goal of the current investigation was to determine whether biosynthesized selenium nanoparticles (Bio-SeNPs) using Azadirachta indica leaves extract might treat mice with Eimeria papillata infection in the jejunal tissue. Five groups of seven mice each were used, as follows: Group 1: Non-infected-non-treated (negative control). Group 2: Non-infected treated group with Bio-SeNPs (0.5 mg/kg of body weight). Groups 3-5 were orally inoculated with 1×103 sporulated oocysts of E. papillata. Group 3: Infected-non-treated (positive control). Group 4: Infected and treated group with Bio-SeNPs (0.5 mg/kg). Group 5: Infected and treated group with the Amprolium. Groups 4 and 5 daily received oral administration (for 5 days) of Bio-SeNPs and anticoccidial medication, respectively, after infection. Bio-SeNPs caused a considerable reduction in oocyst output in mice feces (97.21%). This was also accompanied by a significant reduction in the number of developmental parasitic stages in the jejunal tissues. Glutathione reduced (GSH), glutathione peroxidase (GPx), and superoxide dismutase (SOD) levels were dramatically reduced by the Eimeria parasite, whereas, nitric oxide (NO) and malonaldehyde (MDA) levels were markedly elevated. The amount of goblet cells and MUC2 gene expression were used as apoptotic indicators, and both were considerably downregulated by infection. However, infection markedly increased the expression of inflammatory cytokines (IL-6 and TNF-α) and the apoptotic genes (Caspase-3 and BCL2). Bio-SeNPs were administrated to mice to drastically lower body weight, oxidative stress, and inflammatory and apoptotic indicators in the jejunal tissue. Our research thus showed the involvement of Bio-SeNPs in protecting mice with E. papillata infections against jejunal damage.

The citrus brown mite, Eutetranychus orientalis (Klein), is a significant pest that is widely distributed in different agricultural systems. It causes significant damage to field crops and fruit trees. The most common method of control is the use of chemical acaricides. However, the extensive use of such chemicals developed resistance in addition to environmental and health hazards. Plant-derived acaricides might therefore be an environmentally friendly alternative to synthetic ones in order to increase agricultural production efficiency and to protect consumer health. Following this prospective, the present study was aimed to evaluate the toxic effects of anise (Pimpinella anisum), rosemary (Rosmarinus officinalis), eucalyptus (Eucalyptus globulus), clove (Eugenia caryophyllus), and garlic (Allium sativum) essential oils (EO) against the eggs and adult’s females of E. orentalis under laboratory conditions. Additionally, Euseius scutalis, a predator mite, was investigated as a bioagent to control E. orientalis under field conditions. GC-MS was used to assess the chemical components of the tested EOs. The results showed that anethole; 9, 12-Octadecadienoic acid (Z, Z); camphor; octatriacontyl pentafluoropropiona; and eugenol, were the major components for anise, rosemary eucalyptus, garlic, and cloves, respectively. Comparably, anise has been shown to be the most effective EO against E. orentalis eggs and adults as it exhibited a high level of efficiency and required the lowest concentration to reach the LC50. Additionally, the three Euseius scutalis release rates significantly reduced the Eutetranychus orientalis population. Ten individuals /seedlings in particular induced the highest reduction (84.95%). Based on our findings, we suggest using Euseius scutalis release and anise essential oil as alternative strategies to control of E. orentalis.

Our knowledge of microbial biogeography has advanced in recent years, yet we lack knowledge of the global diversity of some important functional groups. Here, we used environmental DNA from 327 globally collected soil samples to investigate the biodiversity patterns of nitrogen-fixing bacteria by focusing on the nifH gene but also amplifying the general prokaryotic 16S SSU region. Globally, N-fixing prokaryotic communities are driven mainly by climatic conditions, with most groups being positively correlated with stable hot or seasonally humid climates. Among soil parameters, pH, but also soil N content were most often shown to correlate with the diversity of N-fixer groups. However, specific groups of N-fixing prokaryotes show contrasting responses to the same variables, notably in Cyanobacteria that were negatively correlated with stable hot climates, and showed a U-shaped correlation with soil pH, contrary to other N-fixers. Also, the non-N-fixing prokaryotic community composition was differentially correlated with the diversity and abundance of N-fixer groups, showing the often-neglected impact of biotic interactions among bacteria.

Investigations on the parasites that infect migrating birds worldwide are ongoing. To identify the parasite species that infect ardeids, morphological and morphometric characteristics were used. A total of 20 Bubulcus ibis (Ardeidae) specimens were collected from the agricultural lands belonging to the Faculty of Agricultural at Cairo University, which were then examined for nematode parasites. Only one acuariid species, belonging to the Acuariidae family, has been identified, with a prevalence rate of 50 % (10/20) among infected egrets. The Desportesius species isolated from the gizzard of the egret host is morphologically and morphometrically compatible with Desportesius invaginatus, which was previously identified from several Ciconiiformes hosts of Bubulcus ibis (Egypt, India, and Taiwan), Egretta garzetta (France), and Egretta rufescens (USA). Additionally, utilizing the partial small subunit ribosomal RNA (18S rRNA) gene sequence, maximum parsimony based on the Tamura-Nei model was used to infer the phylogeny of the recovered Desportesius species. The query sequences revealed 99.37 % identity for the 18S (MW358651.1) of the previously mentioned D. invaginatus. In addition to clarifying several morphological features of D. invaginatus, this study also provided new DNA data for this species.

Monopisthocotyleans belonging to the family Diplectanidae infect marine fish and are distinguished by a posterior complex haptor. The solider bream Argyrops filamentosus Valenciennes (Perciformes: Sparidae), is one such species that is under consideration caught from the Red Sea (Saudi Arabia). This study is the first description of a diplectanid species from the gills of the soldier bream fish, morphologically by light as well as by molecular analysis of the parasite partial 28S rRNA sequences through multiple alignments and phylogeny by maximum likelihood (ML) analysis which is provided for the first time for the described parasite species. Thirty soldier bream fish were collected from marine water off Saudi Arabia; gills were detached and further analyzed for parasitic infection. A monogenean parasite with a complicated haptor made up of two comparable lamellodiscs, three bilateral pairs of lobes, two pairs of anchors, three bars, and seven pairs of marginal hooks was discovered to naturally infect 21 samples of the examined fish. The molecular analysis of the parasite 28S rRNA and phylogeny revealed a percentage of identities 98.41–77.97 % for Diplectanidae species within a monophyletic clade of Dactylogyridea where a maximum percentage of 98.41 % were obtained for morphologically different sister taxon Lamellodiscus acanthopagri. The results of phylogeny are in line with those obtained through morphological classification, which showed that the parasite identified shared morphological characteristics with Protolamellodiscus senilobatus, a species that had not before been identified by DNA analysis. Under accession number OP419541.1, the obtained parasite sequences were added to the GenBank database.

Giardiasis is considered the most common protozoal infection in humans; it frequently occurs in both developing and developed countries. The present study was designed to investigate the therapeutic and prophylactic effects of Punica granatum peel methanolic extract against experimental giardiasis in comparison to metronidazole (MTZ) in mice infected with Giardia intestinalis. There were five experimental groups, as follows: G1; healthy control group, G2; infected untreated, G3; infected treated with MTZ, G4; Infected treated with pomegranate P. granatum, G5; receiving P. granatum for one week before, during induction of infection than continuous administration for another one week. Giardia cyst per gram stool 7 days and 2 weeks after treatment showed a highly statistically significant difference among the different experimental groups (G3,4,5) versus G2, while the percent reduction of mean Giardia cysts/gm stool 2 weeks after treatment showed a highly significant reduction in G3,4,5 by 92.73, 91.94, and 94.1 %, respectively. The count of Giardia trophozoites in small intestinal content showed a statistically significant difference among G3,4,5 versus G2 respectively with a highly significant reduction in G3,5 and a significant reduction in G4. The percent reduction of mean Giardia trophozoites was approximately 98.69, 58.66, and 95.96 %, respectively. Histopathological examination of small intestines sections of G3 and G4 showed improvement in the pathological lesions including slight restoration of villi/crypt ratio, with partial mucosal ulceration. There was complete healing of intestinal mucosa observable in G5, with a preserved brush border, and no trophozoites were detected. P. granatum peel methanolic extract proved to be valuable in the prevention and treatment of Giardia lamblia infection. Further studies are required to determine the effective therapeutic dose of the extract and its mechanism of action against Giardia lamblia.

Coccidiosis is a parasitic infection threatening poultry products globally. Parasite resistance to drugs is one of the barriers to Eimeria control. Natural products are one of the sources of compounds that prevent parasite infections. The current study was, therefore, conducted to evaluate the effect of Vitis vinifera leaf extract on anti-inflammatory response, oxidative status, and goblet cell response against Eimeria papillate infection in mice. Methanol was used as a solvent for phytochemicals. The mice were divided into six groups: The first group was the control. The second group was uninfected and treated with 200 mg/kg of extract to test toxicity, and the third, fourth, fifth, and sixth groups of mice received 1 × 103 sporulated E. papillate oocysts. The third group received no treatment. The fourth and fifth groups were treated daily with 100 and 200 mg/kg of V. vinifera leaf extract, respectively, while the sixth group received 25 mg/kg of toltrazuril daily via gavage. On day 5 p.i., the animals were sacrificed, and jejunum samples were prepared for analyses of histological sections and oxidative stress. The phytochemical analysis using GC-MS of the extract showed the presence of 12 biologically active compounds. The most effective dose was 200 mg/kg, which significantly decreased the number of parasitic stages in the jejunal sections of the mice. The findings demonstrate that E. papillate infection in mice results in significant histopathological changes in the jejunum, including inflammation, epithelial vacuolation, villi loss, and a decrease in goblet cell density. When infected mice received treatment, the histological injury score within the infected jejunum tissue decreased by 63%, and the goblet cell quantity dramatically increased, approaching the control values. Finally, the extract ameliorated the changes in glutathione and malondialdehyde due to E. papillate infection. The extract was proven to have anti-inflammatory properties and reduce the number of oocysts. Overall, the findings show that V. vinifera leaf extract has significant anticoccidial effects in vivo.

Echinococcosis is a zoonotic disease caused by the genus Echinococcus. Globally, it is one of the most central helminthic diseases. Surgery remains the method of choice to remove cystic Echinococcus. Various sporicidal agents have been used to invalidate the substances in hydatid cysts. Nevertheless, many sporicidal agents cause inflammation and can cause side complications, so their use should be limited. The study aims to evaluate the effectiveness of Vitis vinifera leaf methanolic extract as a sporicidal agent for Echinococcus eggs and protoscolices and determines the best concentration. The mortality and viability of protoscolices were measured in samples exposed to four concentrations of V. vinifera leaf extract (VVLE) (5, 10, 30, and 50 mg/mL) for 5, 10, 20, and 30 min and in eggs exposed to three concentrations (100, 200, and 300 mg/mL) for 24 and 48 h. An infrared spectroscopy chemical test was conducted to assess the presence of numerous expected active components in the extract. The viability of eggs and protoscolices was confirmed using 0.1% eosin staining. Vinifera leaf extract exhibited the decisive sporicidal effect at 100%, 91%, 60%, and 41% after 30 min at concentrations of 50, 30, 10, and 5 mg/mL, and in eggs at 11% and 19% after 24 and 48 h at a concentration of 200 mg/mL, respectively. Increased incubation times and higher dosages often increase mortality. The results exhibited that V. vinifera is effective. This study confirmed that grape leaf extract has high sporicidal activity in vitro. However, more studies are required to determine the exact active chemical and its action mechanism and perform in vivo utilization to confirm these results.

Herbal extracts are promising agents against various parasitic diseases, such as malaria. This study aimed to evaluate the ameliorative action of Eucalyptus camaldulensis extract (ECE) against hepatic damage caused by Plasmodium chabaudi infection. Mice were allocated into five groups as follows: two groups served as the control non-infected groups that received distilled water and ECE, respectively; subsequent three groups were infected with 106 P. chabaudi parasitized erythrocytes; the last two groups were infected with the parasite and then treated with ECE and chloroquine. On day 8 post-infection, the parasite count increased inside erythrocytes (59.4% parasitemia in the infected group). Parasitemia was successfully reduced to 9.4% upon ECE treatment. Phytochemical screening using GC mass spectrometry revealed that ECE contained 23 phytochemical components. Total phenolics and flavonoids in ECE were 104 ± 2 and 7.1± 3 µg/mL, respectively, with 57.2% antioxidant activity. ECE ameliorated changes in liver histopathology and enzymatic activity of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase. In addition, ECE prevented oxidative damage induced by the parasite in the liver, as evidenced by the change in the liver concentrations of glutathione, nitric oxide, malondialdehyde, and catalase. Moreover, ECE was able to regulate the expression of liver cytokines, interleukins-1β and 6, as well as IFN-γ mRNA. ECE possesses antiplasmodial, antioxidant, and anti-inflammatory activity against liver injury induced by the parasite P. chabaudi.

Groupers are large predators in the Serranidae family that feed on fish and small invertebrates. Identification of parasitic taxa of groupers was done concerning their morphological and biological characteristics. A total of 30 Cephalopholis sonnerati (Serranidae) specimens were investigated for digenetic trematodes in the Red Sea, Saudi Arabia. Only one bivesiculid species, belonging to the Bivesiculidae family, has been identified, with a prevalence rate of 60% (18/30) among infected fish. The Bivesicula species obtained from the fish host’s intestine is morphologically and morphometric compatible with Bivesicula claviformis, which was previously identified from Epinephelus fasciatus (Serranidae) in North Borneo. In addition, maximum parsimony based on Tamura-Nei model was utilized to determine the phylogeny of the recovered Bivesicula species using partial small subunit ribosomal RNA gene (18S rRNA) and large subunit ribosomal RNA gene (28S rRNA) sequences. The query sequences showed the identities of 99.39% and 94.02% for 18S (AJ287485.1) and 28S rRNA (AY222182.1) of the previously described Bivesicula claviformis, respectively. The present study demonstrated that Bivesicula claviformis is the first reported of this genus as endoparasites from C. sonnerati, as well as providing novel DNA data for this species.

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.

Carvacrol is a monoterpenoid phenol found in many essential oils that has antibacterial, antifungal and antiparasitic activities. Drug loaded-invasome systems are used to deliver drugs utilizing nanoparticles to improve bioavailability, efficacy, and drug release duration. As a result, the present study developed carvacrol-loaded invasomes and evaluated their acaricidal effect against Rhipicephalus annulatus (cattle tick) and Rhipicephalus sanguineus (dog tick). Carvacrol loaded-invasome (CLI) was prepared and characterized using UV/Vis spectrophotometer, zeta potential measurements, Scanning Transmission Electron Microscopy (STEM), Fourier Transform Infrared (FT-IR) Spectroscopy, and Differential Scanning Calorimetry Analysis. CLI (5%) induced significant mortality (100%) in R. annulatus adult ticks with LC50 of 2.60%, whereas the LC50 of pure carvacrol was 4.30%. Carvacrol and CLI were shown to have a significant larvicidal action on both tick species, with LC50s of 0.24 and 0.21% against R. annulatus and 0.27 and 0.23% against R. sanguineus, respectively. Carvacrol and CLI (5%) induced significant repellent activities for 24 h against R. annulatus and R. sanguineus, as evidenced by the rod method and the petri-dish selective area choice method, respectively. High-performance liquid chromatography (HPLC) demonstrated that the CLI form had 3.86 times the permeability of pure carvacrol. Moreover, carvacrol and CLI inhibited acetylcholinesterase activity and decreased glutathione and malonedealdehyde levels in the treated ticks. In conclusion, invasomes significantly improved adulticidal and repellency activities of carvacrol against both tick species.

Coccidiosis in chickens is one of the major problems in the poultry industry, caused by protozoan parasites of the genus Eimeria. The current study used morphological and molecular characteristics to identify Eimeria spp. infecting domestic chickens (Gallus gallus) in the Riyadh region of Saudi Arabia. In this study, 120 domestic poultry were examined and 30 were found to be infected with oocysts of Eimeria spp. (25%). According to the morphology of the recorded oocysts, five species were found. Eimeria necatrix was the first species discovered, and it was distinguished by oblong, ovoid-shaped oocysts with double-layered walls that measured 20 (23–23) and 17 (16–20) μm. The second species was Eimeria maxima, which had oval- to egg-shaped oocysts with double-layered walls and measurements of 28 (26–29) and 23 (20–24) μm. The third species was Eimeria tenella, characterized by oval-shaped oocysts with double-layered walls and measurements of 21 (20–24) × 17 (16–20) μm. Eimeria praecox was the fourth species that was characterized by spherical-shaped oocysts with single-layered walls and measurements of 21 (19–23) × 20 (19–20) μm. Eimeria acervulina was the last species to have oval-shaped oocysts with double-layered walls and measurements of 20 (18–25) and 17 (14–20) μm. The percentages of infection with Eimeria species were as follows: E. tenella, 10.84%; E. necatrix, 5.84%; E. acervulina, 4.16%; E. maxima, 2.5%; and E. praecox, 1.66%. Nested PCR based on the amplification of internal transcribed spacer I (ITS-I) regions confirmed the presence of the five Eimeria species in the examined fecal samples with their specific amplicon sizes: E. necatrix (383 bp), E. maxima (145 bp), E. tenella (278 bp), E. praecopx (116 bp), and E. acervulina (321 bp).

Objectives: The toxicity of cadmium in various animal models, in particular the liver, has been studied. The objective of this research was to determine the antioxidant and the hepatoprotective role of Pleurotus ostreatus (PE) extract in Cadmium-intoxicated rats. Methods: Rats of the first group were used as control animals while rats of the second group were administered PE for 5 days. Animals of the third group were served as Cadmium intoxicated group and finally, rats of the fourth-group were treated orally with PE afterwards; cadmium was administered over 5 consecutive days. By using atomic absorption, the Cadmium concentration was measured in the liver. Hepatic histological changes were estimated through examination of stained tissue sections. The oxidative status in the liver was investigated through determining the concentration of glutathione (GSH), catalase (CAT), malondialdehyde (MDA) and the reactive oxygen species (ROS) in the liver homogenate. Finally the expression of Nrf2-mRNA was determined using real time PCR. Results: Cadmium induced major hepatic histopathological damage, elevated levels of cadmium, and hepatic oxidative stress where the concentration GSH, CAT, MDA and ROS were affected in the liver. Additionally, cadmium administration resulted in substantial up-regulation of Nrf2-mRNA expression. Moreover, PE attenuated the hepatic cadmium intoxication where it improved the histological impairments and the level of accumulated cadmium in liver tissue was decreased. Likewise, the treatment attenuated the oxidative stress and down-regulated the expression of Nrf2. Conclusions: Our results indicated that PE is associated with significant antioxidant and anti-inflammatory activity and alleviated the hepatotoxicity induced by cadmium chloride.

The parasite, Plasmodium sp. is responsible for Malaria, which kills around half a million people annually and its management relies on efforts of cooperation between health care personnel and the public sector. The effect of Beta vulgaris fresh juice on mice infected by the chloroquine-sensitive Plasmodium berghei berghei through in vivo trials was studied in King Fahad Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia. The EDs were estimated for the fresh juice activity and the secondary screening. There were significant differences in the parasitemia level (p = 0.0) between the negative control group and mice treated by B. vulgaris fresh juice and Chloroquine. The mean parasitemia level in infected untreated mice or B. vulgaris fresh juice was significantly differentiated at concentrations ranging from 10% − 80% (p = 0.0). ED50 and ED90 B. vulgaris fresh juice are estimated to be 24.5 (17.8–33.8) and 40.7 (24–68.8) at 5% confidence limits. This is optimistic and stimulates additional investigation on B. vulgaris fresh juice in vitro trials and further extraction of the active components of B. vulgaris against malaria.

Surra is a parasitic disease caused by the eukaryotic, unicellular hemoprotozoan, Trypanosoma evansi, which affects the development of animal production and is widespread among both domestic and wild animals. As such, in this research, we studied the antiparasitic activity and the ameliorative impact of Eucalyptus camaldulensis leaf extracts (ELE) against T. evansi-induced brain injury and spleen immune response in mice. As a result, we found that ELE decreased the amount of trypanosomes in the blood and improved the weight loss caused by infection. In addition, ELE reduced the parasite-induced brain and spleen histopathological damage. The parasite affected the levels of dopamine and serotonin, but after treatment with ELE, their concentrations significantly decreased to 154 ± 7 and 258 ± 11 μg/g, respectively. We clearly observed the antioxidant activity of ELE because of its ability to increase the induced change in the brain’s total antioxidant capacity and the nitric oxide level. The histopathological changes in the spleen also improved after ELE application. Based on our results, we concluded that ELE possesses antitrypanosomal antioxidant and protective effects in the brains of mice infected with T. evansi. Additional phytochemical screening and molecular studies are required to understand the mechanism underlying the effect of ELE.

Trypanosomiasis is one of the most important diseases affecting economic development for humans and livestock. The current research aimed to assess the in vitro antitrypanosomal effect of methanol extracts of Indigofera oblongifolia leaf (IME) and to examine the brain response after IME treatment of infected mice. In the in vitro method, IME (1.25 mg/ml) was able to kill all parasites after 30 min of exposure. The percentage of the parasite inhibition reached 100% after 180 min of the IME application. Swiss albino mice were infected with 103 Trypanosoma evansi then were daily treated with 100 mg/Kg IME for four days. IME was able to significantly decrease the number of trypanosomes in mice blood. In addition, IME improved the histopathological changes in mice brain induced by the infection. Furthermore, following treatment with IME, the level of epinephrine and nitric oxide in the mice brain homogenate decreased significantly. Taken together, I. Oblongifolia has antitrypanosomal activity and might enhance the brain response to T. evansi.

Objective: Origanum minutiflorum essential oil was evaluated for its chemical components, in vitro antioxidant, scolicidal and anticancer activities. Methods: Oil was extracted and its components were identified with spectrometry and flame ionisation gas chromatography. The oil antioxidant activity was evaluated by DPPH radical scavenging and β-Carotene/linoleic acid bleaching assay. Scolicidal effect of the oil against protoscolices of hydatid cysts was evaluated at different concentrations and exposure times. Also, different oil concentrations were testififed for its anticancer activities on breast cancer cell line (MCF-7), human lung cancer cell line (A-549) and human hepatocellular carcinoma cell line (HepG2) Results: Analyses of the O. minutiflorum oil identified 64 components with carvacrol being the predominant compound. The oil showed a weak free radical scavenging capacity (IC50 at 310 ± 4.2 µg/mL) whilst it was moderately active in term of the suppression of linoleic acid oxidation (64.6%). All the tested oil concentrations, however, revealed a significant scolicidal activity on the protoscoleces of hydatid cysts. The anticancer activity of O. minutiflorum oil was found more in HepG2 cells followed by A-549 and MCF-7 cells and this was evidenced by the LC50 values. Conclusions: The present work strongly suggests that O. minutiflorum could serve as a potential source of scolicidal and anticancer agents.

Coccidiosis has a considerable economic loss in the poultry industry where the parasite Eimeria affect the animal survival. Natural products emerge as an essential and complementary means of controlling avian coccidiosis, in addition to anticoccidial medications. The current study examined the antioxidant activity of Rumex nervosus leaf extract (RE) in chickens infected with Eimeria tenella. RE was able to decrease the induced weight loss due to infection. Also, RE significantly suppressed the oocysts released in chickens’ faeces. Moreover, the oxidative status due to E. tenella infection had been changed after treatment with RE where the level of glutathione, malondialdehyde and nitric oxide was improved indicating the antioxidant activity of the R. nervosus. Our findings suggested that RE could boost the induced oxidative stress in the caecum of infected birds, but studies are needed to determine the mechanism of RE action. Keywords: Rumex nervosus, Coccidiosis, Chickens, Oxidative status, Caecum

Eimerian parasites are the main intestinal tract pathogens in many animals, which invade and damage the intestinal epithelium causing severe injuries and economic loss. Our study was planned to examine the ameliorative effect of Rumex nervosus leaf extracts (RNE) against Eimeria tenella-induced changes in caecal goblet cells and cytokines of chickens. The infected chickens with E. tenella were treated with 50, 100, 200 mg/Kg RNE, respectively. Amprolium was used as a reference drug. Our result showed that RNE contained 8 phytochemical components that were able to decrease the number of oocysts in bird’s faeces. Also, the number of goblet cells was decreased after infection. This number was increased after RNE treatment. In addition, RNE caused upregulation of the mucin gene, MUC2 after E. tenella infection. Moreover, the infection caused upregulation in the inflammatory cytokines IL1β, IL 6, INF-γ and LiTAF. This increase in mRNA expression of IL1β, IL 6, INF-γ and LiTAF was about 5, 5.6, 4 and 5.8 fold, respectively. Collectively, R. nervosus is a promising medicinal plant with anticoccidial, and anti-inflammatory properties and could be used for the treatment of eimeriosis. Keywords: Eimeria tenella, Rumex nervosus, Goblet cells, Inflammation, Cytokines

Abstract Background Vaccination induces survival of otherwise lethal blood-stage infections of the experimental malaria Plasmodium chabaudi. Blood-stage malaria induces extramedullary erythropoiesis in the liver. This study investigates how vaccination affects the course of malaria-induced expression of erythrocytic genes in the liver. Methods Female Balb/c mice were vaccinated at week 3 and week 1 before challenging with 106 P. chabaudi-parasitized erythrocytes. The non-infectious vaccine consisted of erythrocyte ghosts isolated from P. chabaudi-infected erythrocytes. Gene expression microarrays and quantitative real-time PCR were used to compare mRNA expression of different erythrocytic genes in the liver of vaccination-protected and non-protected mice during infections on days 0, 1, 4, 8, and 11 p.i. Results Global transcriptomics analyses reveal vaccination-induced modifications of malaria-induced increases in hepatic gene expression on days 4 and 11 p.i. On these days, vaccination also alters hepatic expression of the erythropoiesis-involved genes Ermap, Kel, Rhd, Rhag, Slc4a1, Gypa, Add2, Ank1, Epb4.1, Epb4.2, Epb4.9, Spta1, Sptb, Tmod1, Ahsp, Acyp1, Gata1, Gfi1b, Tal1, Klf1, Epor, and Cldn13. In vaccination-protected mice, expression of these genes, except Epb4.1, is significantly higher on day 4 p.i. than in un-protected non-vaccinated mice, reaches maximal expression at peak parasitaemia on day 8 p.i., and is slowed down or even decreased towards the end of crisis phase on day 11 p.i.. After day 1 p.i., Epor expression takes about the same course as that of the other erythroid genes. Hepatic expression of Epo, however, is delayed in both vaccinated and non-vaccinated mice for the first 4 days p.i. and is maximal at significantly higher levels in vaccinated mice on day 8 p.i., before declining towards the end of crisis phase on day 11 p.i. Conclusion The present data indicate that vaccination accelerates malaria-induced erythroblastosis in the liver for 1–2 days. This may contribute to earlier replenishment of peripheral red blood cells by liver-derived reticulocytes, which may favour final survival of otherwise lethal blood-stage malaria, since reticulocytes are not preferred as host cells by P. chabaudi.

Mannan oligosaccharide is one of the major components of the yeast cell wall. In the present study, the production, characterization, and bioactivity of yeast mannan were investigated. Four indigenous yeast isolates were obtained from various kinds of juice collected from local markets in Al-Ahsa, KSA, and analyzed using morphological and biochemical methods. Isolate RY1 showed the highest production of mannan. RY1 was identified as Saccharomyces cerevisiae based on morphological characteristics and sequencing of the 18S rRNA gene (GenBank accession number LC479088.1). Mannan-RY1 was characterized by polymer analytical methods, 13C and 1H nuclear magnetic resonance spectroscopy (NMR), and infrared spectroscopy (IR). Interestingly, the mannan extracted from RY1 showed a significant ability to scavenge hydroxyl radicals and superoxide anions. In addition, mannan was found to have pronounced anti-tumor activity against liver (HepG2) and breast cancer (MCF7) cell lines. The results presented in this study confirm the bio-therapeutic activity of S. cerevisiae mannan, which could be used as a potential drug for cancer treatment.

Essential oils (EOs) have recently attracted more interest due to their insecticidal activities, low harmfulness, and rapid degradation in the environment. Therefore, Lavandula steochas (L. steochas) essential oil was assessed for its chemical constituents, in vitro cytotoxicity, and scolicidal, acaricidal, and insecticidal activities. Using spectrometry and gas chromatography, the components of L. steochas EOs were detected. Additionally, different oil concentrations were tested for their anticancer activities when applied to human embryonic kidney cells (HEK-293 cells) and the human breast cancer cell line MCF-7. The oil’s scolicidal activity against protoscolices of hydatid cysts was evaluated at various concentrations and exposure times. The oil’s adulticidal, larvicidal, and repelling effects on R. annulatus ticks were also investigated at various concentrations, ranging from 0.625 to 10%. Likewise, the larvicidal and pupicidal activities of L. steochas against Musca domestica were estimated at different concentrations. The analyses of L. steochas oil identified camphor as the predominant compound (58.38%). L. steochas oil showed significant cytotoxicity against cancer cells. All of the tested oil concentrations demonstrated significant scolicidal activities against the protoscoleces of hydatid cysts. L. steochas EO (essential oil) showed 100% adulticidal activity against R. annulatus at a 10% concentration with an LC50 of 2.34%, whereas the larvicidal activity was 86.67% and the LC50 was 9.11%. On the other hand, the oil showed no repellent activity against this tick’s larva. Furthermore, L. steochas EO achieved 100% larvicidal and pupicidal effects against M. domestica at a 10% concentration with LC50 values of 1.79% and 1.51%, respectively. In conclusion, the current work suggests that L. steochas EO could serve as a potential source of scolicidal, acaricidal, insecticidal, and anticancer agents.

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.

Tick infestation is a serious problem in many countries since it has an impact on the health of animals used for food production and pets, and frequently affects humans. Therefore, the present study aimed to investigate the acaricidal effects of nanoemulsions of essential oils of Melaleuca alternifolia (tea tree, TT) and Citrus limon (lemon oil, CL) against the different stages (adult, eggs, and larvae) of deltamethrin-resistant Rhipicephalus annulatus ticks. Three forms of these oils were tested: pure oils, nanoemulsions, and a binary combination. Tea tree and lemon oil nanoemulsions were prepared, and their properties were assessed using a zeta droplet size measurement and a UV-Vis spectrophotometer. The results showed that TT and CL exhibited higher adulticidal effects in their pure forms than in their nanoemulsion forms, as demonstrated by the lower concentrations required to achieve LC50 (2.05 and 1.26%, vs. 12.8 and 11.4%, respectively) and LC90 (4.01% and 2.62%, vs. 20.8 and 19.9%, respectively). Significant larvicidal activity was induced by the TTCL combination, and LC50 was reached at a lower concentration (0.79%) than that required for the pure and nanoemulsion forms. The use of pure CL oil was found to have the most effective ovicidal effects. In conclusion, pure TT and CL have potent acaricidal effects against phenotypically resistant R. annulatus isolates. It is interesting that the activity levels of TT and CL EOs’ binary and nanoemulsion forms were lower than those of their individual pure forms.

Hysterothylacium species are perhaps the most abundant and diverse group of marine ascaridoids; however, their life cycle and specific identification in larval stages are difficult. In this study, three members of the genus Hysterothylacium (Ward and Magath, 1917) are morphologically described from the digestive tract of Argyrops spinifer, genetically characterized and their relationship with related taxa are compared and discussed. The former species (Hysterothylacium reliquens Norris and Overstreet, 1975) (p = 10%) is mainly characterized by its large body (male 30.89, females 34.15 mm long), the shape of lips, the presence of lateral alae, a short caecum, and a long ventricular appendix, length of spicules, number and distribution of genital papillae, and tail tip with numerous spines. The other species [fourth (L4) and third (L3) larval stages] (p = 46.66, 26.66%, respectively) are mainly characterized by body length (18.21 and 6.20 mm long, respectively), distinct three lips for L4 and poorly developed for L3, a shorter caecum than the ventricular appendix, tail tip lacks any distinct cuticular projections. Molecular tools were done via sequencing and analyzing target regions of ribosomal [internal transcribed spacer (ITS) and small ribosomal DNA (18S)] and mitochondrial [cytochrome c oxidase subunit 1 (COX1)]. These sequences demonstrate discrimination at the species level and confirm the validity of species determined by morphological identification. All sequenced parasite samples were identified as H. reliquens based on nucleotide sequence comparisons with other ascaridoids. In these analyses, Raphidascarididae formed a sister group to Anisakidae with strong nodal support. On MP trees, a genus-specific cluster with a well-support value was delineated in genera Raphidascaris, Contracaecum, and Hysterothylacium. This study demonstrated that selected gene regions of H. reliquens yielded unique sequences (gbl MZ148786.1, MZ148788.1, and MZ148789.1) that confirmed its taxonomic position in Raphidascarididae. Therefore, morphological and molecular tools are important for accurate diagnosis of genus and species of ascaridoids.

There is a growing awareness of the importance of incorporating behavioral changes into infectious disease treatments. In this study, we evaluated whether behavioral changes in mouse brains were altered following infection with the Trypanosoma evansi parasite. Infection significantly reduced locomotor activity as well as vertical and horizontal movements. In the grip strength test, infected mice showed a lower peak than the control group, and in the rotator test, parasite-infected mice spent less time on the rod compared with control mice. The parasite decreased the concentration of both glutathione and catalase in the brain. In addition, the infection caused a significant increase in the levels of malondialdehyde and nitric oxide. The infection induced marked histopathological changes in the brain parts and considerably increased the concentrations of both dopamine and serotonin in the brains of mice. Collectively, T. evansi infection resulted in an oxidative imbalance in the brain. This resulted in neurodegenerative alterations in the brain, as well as a shift in the behavior of mice.

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.

In Egypt, only five species of Dermaptera (earwigs) have been reported. Based on both the morphological and molecular data of the earwig samples collected from a bakery in Beni-Suef, Egypt, we identified the species as Marava arachidis (Spongiphoridae), a cosmopolitan species with no prior records in Egypt. The current study was designed to analyze its predation capability on newly emerged eggs and larvae of the Rhipicephalus annulatus tick. A laboratory functional response study was set up by applying a predation test with various predator-prey ratios as treatments. This experiment was applied using the undefined mix of collected earwigs and the laboratory-collected eggs and the larvae of R. annulatus. The laboratory results showed that the mean number of predated tick eggs was 18.64 ± 11.18 in 24 h under the highest predator-prey ratio (1:10) examined, accompanied by 12.04 ± 4.38 broken but unconsumed eggs. Moreover, M. arachidis predated an average of 12.32 ± 9.07 tick larvae per day. In contrast, the mean dead larvae increased to 38.4 ± 2.30 per day with the highest predator density (1:10). The number of eggs and larvae consumed increased with the predator density. A linear relationship was detected between earwig density and the consumption rates of tick eggs (R2 = 0.99; p = 0.0001) and larvae (R2 = 0.96; p = 0.003). In conclusion, M. arachidis was first recorded in Egypt. This earwig has predation capability on R. annulatus tick eggs and larvae.

Abstract Eimeriosis is a global poultry health problem. In the current study, we investigated the role of Salvadora persica leaf extracts (SE) against murine eimeriosis induced by Eimeria papillata. The infection induced an oocyst output of 6242 ± 731 oocysts/g feces. After treatment with 300 mg⁄kg SE, the oocysts expelled in feces decreased by approximately 3-fold. In addition, the total number of E. papillata in the parasitic stage decreased in the jejunum of mice after treatment with SE. In addition, SE significantly reduced the number of apoptotic cells by approximately 2-fold in the infected jejunum. SE ameliorated the changes in glutathione, malondialdehyde, and catalase due to E. papillata infection. Finally, SE regulated the cytokine genes, interleukin (IL)-1β, IL-6, interferon-γ, and tumor necrosis factor-α, and the apoptotic genes, B-cell lymphoma-2, Bax, and Caspase-3. SE protects the jejunum from E. papillata induced injury and may have potential therapeutic value as a food additive during eimeriosis.

Background: Trypanosomiasis is a neglected tropical disease, transmitted by blood-sucking insects and can affect humans and animals, depending on the species of Trypanosoma parasite. Trypanosoma has acquired resistance to the majority of drugs used; hence, alternative medicines are required. Indigofera oblongifolia leaf extract (IOE) has been shown to treat blood stage malaria. Here, IOE was used to demonstrate its effect on Trypanosoma evansi-infected mice. Methods: Analysis of IOE by gas chromatography–mass spectrometry showed the presence of many active components like flavonoids and phenolics. The mice were divided into three groups as follows: vehicle control, T. evansi-infected mice and T. evansi-infected-treated mice. Results: The findings demonstrate a significant effect of IOE treatment on T. evansi-infected mice. Parasitemia was decreased by 70%, weight loss was reduced, and splenomegaly was significantly decreased. Additionally, IOE improved the histological architecture of the spleen, as shown by the improved histological injury score post-treatment. Anemia was apparent during the course of infection in T. evansi-infected mice; this was reversed upon treatment with IOE to almost the normal level of hemoglobin and erythrocytes. Reduced glutathione and catalase were also ameliorated upon IOE treatment compared to T. evansi-infected mice. Conclusion: Overall, this study shows the ameliorative role of IOE against T. evansi-induced spleen injury in mice. Keywords: Trypanosomiasis, Mouse, Spleen, Indigofera oblongifolia

The green synthesis method is one of the most economic and ecofriendly approach for preparation of metal oxide nanoparticles. In the current study, Moringa oleifera seed was used for synthesis of Ce2O3/M. oleifera (Ce2O3/MNCs) nano-composite. The bio-composite was characterized using FT-IR, XRD, SEM and HR-TEM. The FTIR analysis confirmed the phytochemical involvement in bio-composite. Its crystalline and size was well demonstrated through X-ray Diffraction and HR-TEM. The TEM images revealed these particles in circle shape with average size of 30 nm. The present investigation showed that Ce2O3/MNCs was toxic to B. alexandrina snails with LC50 of 314.5 mg/L. The survival and the reproductive rates of the snails were significantly reduced after exposing to ¼ and ½ of LC50 of Ce2O3/MNCs. The present study showed that Ce2O3/MNCs has significant ovicidal and larvicidal activities. Also, the exposure to ½ of LC50 of Ce2O3/MNCs showed alterations in the tegmental architectures of the head-foot region, in addition it caused significant damages in both of the hermaphrodite and digestive glands of B. alexandrina. Conclusively, Ce2O3/MNCs nano-composite could be utilized as a new molluscicidal agent for the snails of schistosomiasis.

Little information is available about the biodiversity of parasitic copepods in the Arabian Gulf. The present study aimed to provide new information about different parasitic copepods gathered from Parupeneus rubescens caught in the Arabian Gulf (Saudi Arabia). Copepods collected from the infected fish were studied using light microscopy and scanning electron microscopy and then examined using standard staining and measuring techniques. Phylogenetic analyses were conducted based on the partial 28S rRNA gene sequences from other copepod species retrieved from GenBank. Two copepod species, Hatschekia sargi Brian, 1902 and Hatschekia leptoscari Yamaguti, 1939, were identified as naturally infected the gills of fish. Here we present a phylogenetic analysis of the recovered copepod species to confirm their taxonomic position in the Hatschekiidae family within Siphonostomatoida and suggest the monophyletic origin this family. Sequencing the 28S rRNA gene query identified a close relationship of the recovered Hatschekia species to Hatschekia japonica (gb| KR048858.1). This study identifies a new host for Hatschekia species isolated from Saudi Arabia and performs the first molecular analysis of their 28S rRNA genes.

Malaria is an important health problem in subtropical and tropical areas around the world. Infection with protozoan parasites of the Plasmodium genus, which grow inside host erythrocytes, causes malaria and may lead to morbidity and mortality. Liver tissue plays an important role in the pathogenesis of malaria and is closely involved in parasitic pre-erythrocytic development. Numerous published studies have demonstrated that the liver is not only the source of Plasmodium parasites prior to erythrocytic growth but is also a primary immune effector toward the blood stage of the malaria life cycle. Despite efforts to improve antimalarial drugs and vaccines, Plasmodium species that cause severe malaria are being detected increasingly frequently in endemic regions. In this study, Salvia officinalis (S. officinalis) leaf extract was employed to synthesize silver nanoparticles (Ag-NPs). This method is eco-friendly and represents a single-step technique for the biosynthetic process; therefore, it has attracted considerable attention. Accordingly, we biosynthesized Ag-NPs with extract of the S. officinalis leaf and examined the antimalarial activity of these nanoparticles in a murine model of Plasmodium chabaudi malaria (P. chabaudi malaria). Forty mice were chosen and classified into four types: infected group, healthy control, pretreated mice infected after treatment with 50 mg/kg of S. officinalis leaf extract-biosynthesized Ag-NPs for two weeks, and post-treated mice infected before treatment with 50 mg/kg of S. officinalis leaf extract-biosynthesized Ag-NPs (administered daily for 7 d). In this study, both pre-treatment and post-treatment with Ag-NPs produced a substantial reduction in parasitemia relative to the infected group. We investigated the antiplasmodial and hepatoprotective effects of S. officinalis leaf extract-biosynthesized Ag-NPs on P. chabaudi-induced inflammation and hepatic oxidative stress markers.

Globally, the economic losses due to hard ticks infestation and the control of the associated diseases have been calculated at USD $13.9-18.7 billion per year. The economic impact is related to its direct damage to the skins, blood loss, anemia, severe immunological reactions and indirect losses that related to the effects of hemoparasites, cost of treatment for clinical cases and expenses incurred in the control of ticks. The current study evaluated the acaricidal activities of fennel Foeniculum vulgare essential oil and its main components; trans-anethole and fenchone; against R. annulatus. GC-MS analysis revealed that this oil contained 16 components representing 99.9% of the total identified compounds with E-anethole being the predominant component(64.29%), followed by fenchone (9.94%). The fennel oil and trans-anethole showed significant acaricidal activities. The LC50 of the fennel oil was attained at concentrations of 12.96% for adult ticks and 1.75% for tick larvae meanwhile the LC50 of trans-anethole was reached at concentrations of 2.36% for adult tick and 0.56% for tick larvae. On the contrary, fenchone showed no any significant adulticidal activities and its LC50 attained at a concentration of 9.11% for tick larvae. Regarding repellence activities, trans-anethole achieved 100% repellency at the concentration of 10% while fennel showed 86% repellency at the same concentration. Fenchone showed no repellency effect. Treatment of larvae with fennel, trans-anethole, and fenchone LC50 concentrations significantly inhibited the acetylcholinesterase activity. Meanwhile, glutathione s-transferase activity was significantly decreased in fennel treated larvae but no significant effect was found in the larvae of trans-anethole and fenchone groups. These results indicate that the acaricide effect of fennel oil may attributed to its high content of trans-anethole. This was supported by potent adulticidal, larvicidal, and repellency effects of trans-anethole against Rhipeciphalus annulatus tick and therefore it could be included in the list of acaricide of plant origin.

Abstract Natural products are ecofriendly agents that can be used against parasitic diseases. Eimeria species cause eimeriosis in many birds and mammals and resistance to available medications used in the treatment of eimeriosis is emerging. We investigated the in vitro and in vivo activity of Morus nigra leaf extracts (MNLE) against sporulation of oocysts and infection of mice with Eimeria papillata. Phytochemical analysis of MNLE showed the presence of seven compounds and the in vitro effects of MNLE, amprolium, DettolTM, formalin, ethanol, and phenol were studied after incubation with oocysts before sporulation. Furthermore, infection of mice with E. papillata induced an oocyst output of approximately 12 × 105 oocysts/g of feces. MNLE significantly decreased oocyst output to approximately 86% and the total number of parasitic stages in the jejunum by approximately 87%. In addition, the reduction in the number of goblet cells in the jejuna of mice was increased after treatment. These findings suggest that mulberry exhibited powerful anticoccidial activity.