Pharmacological Effects of Grape Leaf Extract Reduce Eimeriosis-Induced Inflammation, Oxidative Status Change, and Goblet Cell Response in the Jejunum of Mice.

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.

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

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

In vivo anticoccidial, antioxidant, and anti-inflammatory activities of avocado fruit, Persea americana (Lauraceae), against Eimeria papillata infection.

Apicomplexan parasites, especially Eimeria sp., are the main intestinal murine pathogens, that lead to severe injuries to farm and domestic animals. Many anticoccidial drugs are available for coccidiosis, which, leads to the development of drug-resistant parasites. Recently, natural products are considered as an alternative agent to control coccidiosis. This study was designed to evaluate the anticoccidial activity of the Persea americana fruit extract (PAFE) in male C57BL/6 mice. A total of 35 male mice were divided into seven equal groups (1, 2, 3, 4, 5, 6, and 7). At day 0, all groups except the first group which served as uninfected-untreated control were infected orally with 1 × 103E. papillata sporulated oocysts. Group 2 served as uninfected-treated control. Group 3 was considered an infected-untreated group. After 60 min of infection, groups 4, 5, and 6 were treated with oral doses of PAFE aqueous methanolic extract (100, 300, and 500 mg/kg of body weight, respectively). Group 7 was treated with amprolium (a reference drug for coccidiosis). PAFE with 500 mg/kg, was the most effective dose, inducing a significant reduction in the output of oocysts in mice feces (by about 85.41%), accompanied by a significant decrease in the number of the developmental parasite stages and a significant elevation of the goblet cells in the jejunal tissues. Upon treatment, a significant change in the oxidative status due to E. papillata infection was observed, where the levels of glutathione (GSH) increased, while, levels of malondialdehyde (MDA) and nitric oxide (NO) were decreased. In addition, the infection significantly upregulated the inflammatory cytokines of interleukin-1ß (IL-1ß), tumor necrosis factor-alpha (TNF-α), and interferon-γ (IFN-γ). This increase in mRNA expression of IL-1ß, TNF-α, and IFN-γ was about 8.3, 10.6, and 4.5-fold, respectively, which significantly downregulated upon treatment. Collectively, P. americana is a promising medicinal plant with anticoccidial, antioxidant, and anti-inflammatory activities and could be used for the treatment of coccidiosis.

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

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

Hepatoprotective activity of Eucalyptus camaldulensis extract in murine malaria mediated by suppression of oxidative and inflammatory processes.

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-1ß 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.

Molecular identification of the rodent-borne pathogen Rodentolepis nana using the genetic markers of ITS-1, 18 S, and 28 S rDNA.

BACKGROUND: Rodentolepis nana (syn. Hymenolepis nana), the most common cyclophyllid tapeworm infecting rodents, is a well-studied gastrointestinal parasite in mice and belongs to the family Hymenolepididae. METHODS: The present study focuses on the molecular analysis for the nuclear genes (ITS-1, 18 S, and 28 S rDNA) used for the accurate recognition of the recovered Rodentolepis species. RESULTS: The annotated partial ITS-1, 18 S, and 28 S rDNA gene regions were deposited in GenBank (gbǀ MW310394.1, gbǀ MW327585.1, and gbǀ MW324479.1, respectively) and further used in the maximum likelihood method (ML) to clarify their genetic relationships at the species level. The interrogation sequence of R. nana was aligned and belonged to the family Hymenolepididae, in the same group as all Hymenolepis species, which were distinct from Cyclophyllidea cestodes, especially species belonging to Anoplocephalidae and Taeniidae. Sequence data support the paraphyly of Hymenolepis species. CONCLUSIONS: The phylogeny supports the availability of the ITS-1, 18 S, and 28 S rDNA genes as reliable genetic markers for evolutionary relationships.

Survey of Dicrocoelium dendriticum infection in imported Romani and local sheep (Ovis aries), and potential epidemiological role in Saudi Arabia.

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.

Heteromicrocotyla polyorchis Unnithan, 1961 (Monogenea: Heteromicrocotylidae), a gill parasite of the yellow-spotted trevally, Carangoides fulvoguttatus (Carangidae) from Saudi Arabia: Morphology and phylogeny.

Little information for parasitic infections of Carangoides fulvoguttatus was recorded. The present study was intended to investigate the gill parasite Heteromicrocotyla polyorchis of this fish and to provide a full morphological description and clarify its taxonomic status through phylogenetic analysis of the 28S rRNA gene region. A total of sixty fish specimens have been collected from the studied area (the Red Sea in Jeddah Province, Saudi Arabia) and gills were isolated and examined for identification of parasites. Using light electron microscopy, the recovered monogenean parasite's morphology was exhaustively characterized and described. Microscope examinations found that this parasite species represent Heteromicrocotyla polyorchis, and it could be distinguished from congeners of the same genus by armed genital atrium and cirrus sac, follicular post-ovarian testes, unique shape and number of clamps on both haptor sides, and the dorsally curved tip of the male copulatory organ. Morphological features were combined with molecular analysis of the 28S rRNA gene region. The selected gene for the isolated Heteromicrocotyla species was analyzed using appropriate primers to assist in phylogeny with those in the GenBank database. The present monogenean species was characterized by unique genetic sequences that were analyzed and deposited in the GenBank for the first time under the accession number MW406473. Phylogenetic analyses reported that the maximum identity between the current Heteromicrocotyla species and taxa of Heteromicrocotylidae was between 91.42 and 92.09% and confirmed its taxonomic status in this family with a well-distinct clade. The present study supported the second report of H. polyorchis as carangid fish ectoparasites and investigates the first appearance in C. fulvoguttatus inhabiting Saudi Arabia.

Cucullanus bulbosus (Lane, 1916) Barreto, 1918 (Nematoda, Cucullanidae) from the common ponyfish Leiognathus equulus (Leiognathidae): Morphology and molecular study.

The common ponyfish Leiognathus equulus is a marine fish species with very high commercial value. Little information is available about its parasitic infections. Based on light and scanning electron microscopy, as well as sequencing and analysis of the partial regions of the ITS-1, 18S rRNA, COX1 genes, were employed for the systematic evaluation of a nematode parasite, which it first isolated from L. equulus in Jeddah Province, Red Sea, Saudi Arabia. Results revealed that this nematode parasite closely resembles the previously described Cucullanus bulbosus. Microscopic examination showed that it distinguished from congeners by the unique structure of hemispherical elevation at pseudobuccal capsule level, the ratio of esophagus/body length, spicules size, presence of pre-cloacal sucker, rod-shaped gubernaculum, and the arrangement of caudal papillae in males. Phylogenetic analyses based on ITS-1, 18S rRNA, and COX1 gene regions were constructed to investigate phylogenetic relationships between this parasite species and other related taxa. Results supported that Cucullanus bulbosus resembles a sister of Cucullanus genypteri, Cucullanus pulcherrimus, Cucullanus bourdini, Cucullanus extraneus, and Cucullanus hainanensis by using different genetic markers. This study provides more information about combining morphological and molecular data to identify Cucullanus species with the first natural occurrence in the common ponyfish inhabited in Saudi Arabia.

Neuro-biochemical changes induced by zinc oxide nanoparticles.

Nanoparticles are now widely used in various aspects of life, especially zinc oxide nanoparticles (ZnNPs) that used in mouth washing, cosmetics, sunscreens, toothpaste and root canal flings. This research aims to determine the impact of ZnNPs on healthy mice's brain tissue. ZnNPs have caused major changes in the brain monoamines (dopamine, norepinephrine and serotonin) and ions such as Ca2+, Na+, K+ and Zn2+. Concerning the histological picture, administration of ZnNPs caused some histopathological impairment in brain tissue. In addition, ZnNPs reduced the level of glutathione and catalase in brain tissue, although an increase in the level of nitrite / nitrate and ROS was observed, while the level of malondialdhyde was not significantly altered. Moreover, ZnNPs induced DNA fragmentation in brain of mice. Collectively, the obtained results revealed that ZnNPs affected the brain levels of investigated monamines, ions, enzymatic and non-enzymatic antioxidants thus they may have potential influence on central nervous system.

Biosynthesized silver nanoparticles regulate the iron status in the spleen of Plasmodium chabaudi-infected mice.

Malaria is a dangerous disease affecting millions around the globe. Biosynthesized nanoparticles are used against a variety of diseases including malaria worldwide. Here, silver nanoparticles (AgNPs) synthesized from the leaf extracts of Indigofera oblongifolia have been used in the treatment of mice infected with Plasmodium chabaudi to evaluate the expression of iron regulatory genes in the spleen. Infrared spectroscopy was used to identify the expected classes of compounds in the extract. AgNPs were able to decrease the parasitemia nearly similar to the used reference drug, chloroquine. In addition, AgNPs significantly decreased the spleen index after infection. Moreover, the iron distribution was increased after the treatment. Finally, AgNPs could regulate the mice spleen iron regulatory genes, Lipocalin 2 (Lcn2), transferrin receptor 1 (TFR1) and hepcidin antimicrobial peptide (Hamp). Taken together, our findings indicate that AgNPs have antimalarial activity and can control the state of iron in spleen. We need further investigations to determine mechanisms of action of the AgNPs.