Linalool-zinc oxide nanocomposite controls Toxoplasma gondii infection through inhibiting inflammation, oxidative stress, and pathogenicity.

The present in vitro and in vivo study aimed to fabricate and characterize linalool-zinc oxide nanoparticles (Lin-ZNP) and evaluate their effectiveness against Toxoplasma gondii infection in terms of inflammation, oxidative stress, and pathogenicity. Lin-ZNP was synthesized using an ethanolic solution of polyvinyl alcohol. The anti-Toxoplasma and cytotoxicity activities of Lin-ZNP were investigated, along with its effects on nitric oxide (NO) production, caspase-3 activity, and pro-inflammatory genes. After treating T. gondii-infected mice with Lin-ZNP for 14 days, the number and size of tissue cysts, antioxidant potential, pro-inflammatory cytokines, and T. gondii pathogenicity-related genes were evaluated by real-time polymerase chain reaction and Western blot analysis. The Lin-ZNP composite showed a reduced tendency with an average size of 105 nm. Lin-ZNP significantly reduced the viability of tachyzoites. The obtained selectivity index higher than 10, indicating high specificity for parasites with low cytotoxicity to normal cells. The Lin-ZNP significantly (p < 0.05) increased the production of NO, caspase-3 activity, and the expression levels of pro-inflammatory genes. Lin-ZNP significantly (p < 0.001) decreased the size and number of tissue cysts and caused a significant reduction in the level of malondialdehyde and a considerable increase (p < 0.001) in antioxidant enzymes and their expression genes. Lin-ZNP significantly downregulated both mRNA and protein expression of the inflammation-related markers associated with the TLRs/NF-κB pathway. The expression levels of the T. gondii pathogenicity-related genes were significantly downregulated (p < 0.05). The recent survey indicated that Lin-ZNP manages T. gondii infection by its antioxidant activity and inhibiting the TLRs/NF-κB pathway without toxicity in mice.

First report of Kudoa species (Myxozoa, Multivalvulida) infection in purple-spotted Bigeye (Priacanthus tayenus) from the Saudi Arabian Gulf.

The purple-spotted bigeye, Priacanthus tayenus, is a marine benthic fish native to the Indian and Pacific Oceans, including the Arabian Gulf in Saudi Arabia. This study identified a myxozoan parasite infecting wild P. tayenus from the Saudi Arabian Gulf. These parasites produced spherical to ovoid-shaped, white plasmodia enclosed within pseudocysts in the fish musculature. The annual infection rate was 5.1%, with the highest prevalence in summer (7.6%), followed by spring (6%), and autumn (2.5%), while no infections were observed in winter. The number of plasmodia per fish ranged from 100 to 150 (135.1 ± 16.2). Their dimensions were 4-4.7 mm (4.3 ± 0.3 mm) in length and 4.5-7 mm (6 ± 1.1 mm) in width. Milky-colored exudates within the plasmodia contained mature spores measuring 8-9 µm (8.6 ± 0.4 µm) x 6-7.5 µm (6.9 ± 0.5 µm). The polar capsules of the spores exhibited dimensions of 2-5 µm (3.5 ± 0.5 µm) x 2.5-4.5 µm (3 ± 0.45 µm). Both morphological and genetic analyses confirmed these plasmodia as a novel Kudoa species. Histopathological examination revealed atrophy in the surrounding muscles without an inflammatory response. This study documents the first occurrence of a novel Kudoa sp. in P. tayenus at the Jubail landing site in Saudi Arabia, emphasizing the need for further surveillance and investigations to elucidate its pathogenesis and implications for wild fish stocks.

Cross-sectional community-based study to assess the awareness of toxoplasmosis in Saudi Arabia.

A few reports have been published and documented low level of awareness on toxoplasmosis among Saudi women. Herein, a cross sectional community based study was undertaken to evaluate basic knowledge on toxoplasmosis among residents in the Eastern province (Sharqiyah). Thisstudy was conducted between December 2022 and January 2023 on 334 females from different ages and educational backgrounds. Analysis of their responses revealed that only (24.9%) had heard about the disease. However, (69.8%) properly identified cats as the source of Toxoplasma gondii (T. gondii), but a smaller percentage (47.7%) knew that they might become infected through handling cat feces, and a few (26.3%) believed that bad hand hygiene can result in T. gondii infection. A few males (n = 26) have also participated, for the first time in Saudi Arabia, and displayed also low level of knowledge on toxoplasmosis. We do recommend establishing educational programs for females, in various Saudi provinces, to raise awareness on toxoplasmosis.

Camellia sinensis Aqueous Extract: A Promising Candidate for Hepatic Eimeriosis Treatment in Rabbits.

Eimeria stiedae (E. stiedae) is a common coccidian species that infects the liver and causes economic losses for the rabbit industry. This study aimed to determine the efficiency of green tea aqueous extract (GTE) as a natural treatment for eimeriosis caused by E. stiedae. Male rabbits Cuniculus L. (Oryctolagus) of the New Zealand White rabbit strain (4-4.5 months) were used, as they are suitable for research and conducting experiments. Thirty rabbits were allocated into six groups, with five rabbits in each group; the G1 group (non-infected untreated) served as a negative control group; the G2 group was not infected and treated with 250 mg GTE; the G3 group was not infected and treated with 500 mg GTE; the G4 group was untreated and was infected with 3 × 104 Sporulated E. stiedae oocysts, which served as a positive control group; the G5 group was infected and treated with 250 mg GTE; and the G6 group was infected and treated with 500 mg GTE. The hematological and biochemical analyses of each group of rabbit sera were carried out. Phytochemical analysis was performed to evaluate the active components in GTE leaves using the following methods: IR spectroscopy, liquid chromatography-mass spectrometry (LC-MS), energy-dispersive X-ray spectroscopy (EDX), and scanning electron microscopy. The infected rabbit groups treated with GTE at both doses of 250 and 500 mg/kg exhibited a significant decrease in the extent of E. stiedae oocyst shedding compared with the infected untreated group at 14, 21, and 28 days post-infection. Also, treatment with green tea showed improvement in liver weight compared with the enlarged livers of infected, untreated rabbits. The disturbance in serum liver enzymes' gamma-glutamyl transferase (GGT) and aspartate aminotransferase (AST/GOT) levels, as well as serum glucose, potassium, uric acid, cholesterol, and urea levels, were improved after the treatment of infected rabbit groups with green tea compared with the infected untreated group. Moreover, in this study, the images of the egg stages of the parasite were taken using a fluorescence microscope at 25 µm and 26 µm magnifications. This study provides promising results for the effective cell absorption of the aqueous extract of green tea, which was confirmed in the analyzed images using a scanning electron microscope at 5 µm and 20 µm magnifications.

Antiparasitic activity of FLLL-32 against four Babesia species, B. bovis, B. bigemina, B. divergens and B. caballi, and one Theileria species, Theileria equi in vitro, and Babesia microti in mice.

Introduction: FLLL-32, a synthetic analog of curcumin, is a potent inhibitor of STAT3's constitutive activation in a variety of cancer cells, and its anticancer properties have been demonstrated both in vitro and in vivo. It is also suggested that it might have other pharmacological activities including activity against different parasites. Aim: This study therefore investigated the in vitro antiparasitic activity of FLLL-32 against four pathogenic Babesia species, B. bovis, B. bigemina, B. divergens, and B. caballi, and one Theileria species, Theileria equi. In vivo anti-Babesia microti activity of FLLL-32 was also evaluated in mice. Methods: The FLLL-32, in the growth inhibition assay with a concentration range (0.005-50 µM), was tested for it's activity against these pathogens. The reverse transcription PCR (RT-PCR) assay was used to evaluate the possible effects of FLLL-32 treatment on the mRNA transcription of the target B. bovis genes including S-adenosylhomocysteine hydrolase and histone deacetylase. Results: The in vitro growth of B. bovis, B. bigemina, B. divergens, B. caballi, and T. equi was significantly inhibited in a dose-dependent manner (in all cases, p < 0.05). FLLL-32 exhibits the highest inhibitory effects on B. bovis growth in vitro, and it's IC50 value against this species was 9.57 µM. The RT-PCR results showed that FLLL-32 inhibited the transcription of the B. bovis S-adenosylhomocysteine hydrolase gene. In vivo, the FLLL-32 showed significant inhibition (p < 0.05) of B. microti parasitemia in infected mice with results comparable to that of diminazene aceturate. Parasitemia level in B. microti-infected mice treated with FLLL-32 from day 12 post infection (pi) was reduced to reach zero level at day 16 pi when compared to the infected non-treated mice. Conclusion: The present study demonstrated the antibabesial properties of FLLL-32 and suggested it's usage in the treatment of babesiosis especially when utilized in combination therapy with other antibabesial drugs.

Babesia microti alleviates disease manifestations caused by Plasmodium berghei ANKA in murine co-infection model of complicated malaria.

Malaria remains one of the most significant health issues worldwide, accounting for 2.6% of the total global disease burden, and efforts to eliminate this threat continue. The key focus is to develop an efficient and long-term immunity to this disease via vaccination or therapeutic approach, and innovative strategies would enable us to achieve this target. Previously, using a mouse co-infection disease model, cross-protection was illustrated between Babesia microti and Plasmodium chabaudi. Hence, this study was planned to elucidate the impact of acute B. microti Peabody mjr and Plasmodium berghei ANKA co-infection on the consequence of complicated malaria in the C57BL/6J mouse model of malaria. Furthermore, immune response and pathological features were analyzed, and the course of the disease was compared among experimental groups. Our study established that acute B. microti infection activated immunity which was otherwise suppressed by P. berghei. The immunosuppressive tissue microenvironment was counteracted as evidenced by the enhanced immune cell population in co-infected mice, in contrast to P. berghei-infected control mice. Parasite sequestration in the brain, liver, lung, and spleen of co-infected mice was significantly decreased and tissue injury was ameliorated. Meanwhile, the serum levels of IFN-γ, TNF-α, and IL-12p70 were reduced while the secretion of IL-10 was promoted in co-infected mice. Eventually, co-infected mice showed an extended rate of survival. Hereby, the principal cytokines associated with the severity of malaria by P. berghei infection were TNF-α, IFN-γ, and IL-12p70. Moreover, it was evident from our flow cytometry results that innate immunity is crucial and macrophages are at the frontline of immunity against P. berghei infection. Our study recommended further investigations to shed light on the effects of babesiosis in suppressing malaria with the goal of developing Babesia-based therapy against malaria.

Development of a promising antigenic cocktail for the global detection of Babesia caballi in horse by ELISA.

In this study, we designed novel truncated Babesia caballi (B. caballi) recombinant proteins from the previously used B. caballi proteins; 134-Kilodalton Protein (rBC134) and Merozoite Rhoptry 48 Protein (rBC48). Then, we evaluated the diagnostic performance of the newly designed proteins when used as a single antigen or when used as cocktail antigen consists of rBC134 full length (rBC134f) + newly designed rBC48 (rBC48t) or newly designed rBC134 (rBC134t) + rBC48t for the detection of B. caballi infection in horse using indirect enzyme-linked immunosorbent assay (iELISA). We used one dose and a half of each antigen in the cocktail formulas. The serum samples were collected from different endemic areas in addition to the sera collected from horses experimentally infected with B. caballi were used in the present study. Cocktail antigen in full dose of (rBC134f + rBC48t) exhibited the highest optical density (OD) values with B. caballi-infected sera and showed the lowest OD values with normal equine sera or B. caballi, and Theileria equi mixed infected sera in comparison with the single antigen. Interestingly, the same cocktail antigen exhibited the highest concordance rate (76.74%) and kappa value (0.79) in the screening of 200 field serum samples collected from five B. caballi endemic countries, including South Africa (n = 40), Ghana (n = 40), Mongolia (n = 40), Thailand (n = 40), and China (n = 40) using iELISA and the results were compared to those of indirect fluorescent antibody test (IFAT) as a reference. Moreover, the identified promising cocktail full dose antigen (rBC134f + rBC48t) showed that it can detect the infection as early as the 4th day post-infection in sera collected from experimentally infected horses. The obtained results revealed the reliability of the rBC134f + rBC48t cocktail antigen when used in full dose for the detection of specific antibodies to B. caballi in horses which will be useful for epidemiological surveys and control of equine babesiosis.

Gastrointestinal Parasites of Dogs in Egypt: An Update on the Prevalence in Dakahlia Governorate and a Meta-Analysis for the Published Data from the Country.

Since the last survey on gastrointestinal (GIT) parasites infecting dogs in Dakahlia governorate, Egypt, was published 40 years ago, the present study detected various GIT parasites in feces of 78 stray dogs in this governorate. Twenty-one dogs (35.9%) had eggs/oocysts of eight different parasites including Toxocara canis (19.2%), Toxascaris leonina (2.6%), hookworms (1.3%), Taenia species (5.1%), Dipylidium caninum (2.6%), Cystoisospora canis (5.1%), Cystoisospora ohioensis (2.6%), and Neospora caninum-like oocysts (1.3%). These results were combined in various meta-analyses with findings of all published surveys on GIT parasites of dogs in Egypt to underline the potential parasitic zoonoses from dogs in the country. Feces and/or gastrointestinal tracts of 19,807 dogs from various Egyptian governorates, but particularly Cairo, have been microscopically tested in 182 datasets published between 1938 and 2022, revealed during our systematic database search. Toxocara canis, interestingly, displayed a twofold higher pooled prevalence (24.7%) when compared to the published global pooled prevalence for T. canis, indicating that dogs represent a major risk for toxocariasis in humans from Egypt. Dipylidium caninum (25.4%) as well as various Taenia species (17.1%) also displayed high pooled prevalences. On the contrary, lower pooled prevalence was estimated for the most important zoonotic taeniid "Echinococcus granulosus" (2.4%) as well as for hookworms (1.8%) in comparison to what has been published from other countries in the region. Relatively high prevalences were estimated for three protozoa detected in dogs and are common to infect children in Egypt; Cryptosporidium (5.5%), Giardia (7.4%), and Entamoeba histolytica (9.8%). In general, the pooled prevalence estimated for various parasites detected in dogs from Egypt has decreased in the recent years, sometimes by as much as one-fifth, but this great decline is statistically insignificant, which should alert the veterinary and public health authorities to continue their efforts for controlling these parasites in a "One Health" approach.

Protective Effect of Litchi chinensis Peel Extract-Prepared Nanoparticles on Rabbits Experimentally Infected with Eimeria stiedae.

The present study used Litchi chinensis peel extract to synthesize silver nanoparticles (AgNPs). This technique is eco-friendly and can be performed in a single step; thus, it has attracted great attention for NPs biosynthesis. Herein, we biosynthesized AgNPs with L. chinensis peel extract and examined their anticoccidial activity in rabbit hepatic coccidiosis induced by E. stiedae infection. Thirty-five rabbits were allocated into seven groups: a healthy group (G1), an infected control group (G2), four groups infected before treatment with 10 mg/kg L. chinensis peel extract-biosynthesized AgNPs (G3, G5) or 50 mg/kg amprolium (G4, G6), and rabbits infected after two weeks of pretreatment with 10 mg/kg L. chinensis eel extract-biosynthesized AgNPs (G7). In this study, both pre-and post-treatment with AgNPs produced a substantial reduction in fecal oocyst output, liver enzyme levels, and histopathological hepatic lesions relative to the infected group. In conclusion, L. chinensis peel extract-prepared AgNPs should be considered harmless and efficient in the cure of hepatic coccidiosis in rabbits.

Repurposing of the Malaria Box for Babesia microti in mice identifies novel active scaffolds against piroplasmosis.

BACKGROUND: An innovative approach has been introduced for identifying and developing novel potent and safe anti-Babesia and anti-Theileria agents for the control of animal piroplasmosis. In the present study, we evaluated the inhibitory effects of Malaria Box (MBox) compounds (n = 8) against the growth of Babesia microti in mice and conducted bioinformatics analysis between the selected hits and the currently used antibabesial drugs, with far-reaching implications for potent combinations. METHODS: A fluorescence assay was used to evaluate the in vivo inhibitory effects of the selected compounds. Bioinformatics analysis was conducted using hierarchical clustering, distance matrix and molecular weight correlation, and PubChem fingerprint. The compounds with in vivo potential efficacy were selected to search for their target in the piroplasm parasites using quantitative PCR (qPCR). RESULTS: Screening the MBox against the in vivo growth of the B. microti parasite enabled the discovery of potent new antipiroplasm drugs, including MMV396693 and MMV665875. Interestingly, statistically significant (P < 0.05) downregulation of cysteine protease mRNA levels was observed in MMV665875-treated Theileria equi in vitro culture in comparison with untreated cultures. MMV396693/clofazimine and MMV665875/atovaquone (AV) showed maximum structural similarity (MSS) with each other. The distance matrix results indicate promising antibabesial efficacy of combination therapies consisting of either MMV665875 and AV or MMV396693 and imidocarb dipropionate (ID). CONCLUSIONS: Inhibitory and hematology assay results suggest that MMV396693 and MMV665875 are potent antipiroplasm monotherapies. The structural similarity results indicate that MMV665875 and MMV396693 have a similar mode of action as AV and ID, respectively. Our findings demonstrated that MBox compounds provide a promising lead for the development of new antibabesial therapeutic alternatives.

Molecular Epidemiology and Species Diversity of Tick-Borne Pathogens of Animals in Egypt: A Systematic Review and Meta-Analysis.

Ticks and tick-borne pathogens (TTBPs) are listed among the most serious concerns harming Egyptian livestock's productivity. Several reports on tick-borne pathogens (TBPs) from various geographical regions in the country were published. However, data on the molecular characterization of TBPs are the most beneficial for understanding the epidemiology of this important group of pathogens. In this study, we present the first meta-analysis on the molecular epidemiology and species diversity of TBPs infecting animals in Egypt. All published studies on TBPs were systematically collected from various databases (PubMed, Scopus, ScienceDirect, the Egyptian Knowledge Bank, and Google Scholar). Data from eligible papers were extracted and subjected to various analyses. Seventy-eight studies were found to be eligible for inclusion. Furthermore, ticks infesting animals that were molecularly screened for their associated pathogens were also included in this study to display high species diversity and underline the high infection risk to animals. Theileria annulata was used as parasite model of TBPs to study the genetic diversity and transmission dynamics across different governorates of Egypt. This study extends cross-comparisons between all published molecular data on TBPs in Egypt and provides resources from Egyptian data in order to better understand parasite epidemiology, species diversity, and disease outcome as well as the development and implementation of prevention and control methods for public health, veterinary care practitioners, and animal owners all over the country.

Identification of Theileria spp. in sheep and goats from Jeddah, Saudi Arabia, using molecular techniques.

BACKGROUND: Thileriosis is a tick -born disease caused by hemoprotozoan parasites which has global veterinary and economic implications. METHODS: Blood samples were collected from 216 sheep and 83 goats from Jeddah, Saudi Arabia, were analyzed to determine whether the animals were infected with Theileria spp. parasites. The parasites were detected using a polymerase chain reaction (PCR) targeting the gene of 18S rRNA followed by sequencing. RESULTS: According to obtained findings, Theileria spp. were detected in sheep (57.8%, 48/83) and goats (51.9%, 112/216). Phylogenetic analysis to sequence data showed that T. ovis identified in this study were found to be closely connected to an isolate from Turkey, with 84.4-99.8% pairwise identity and 52.35-99.79% coverage.

Prevalence of head lice (Pediculus humanus capitis) infestation among schools workers in the Eastern Region, Saudi Arabia.

In a cross-sectional survey in February-May 2019, the prevalence of Pediculosis capitis with demographic data and the behavioral practices were investigated among 750 participants in the Eastern region of Saudi Arabia. Female participation was highly remarked with a percentage of 94.08% compared to that of male one about 5.91%. A deficiency of knowledge about lice infestation was noted especially among illiterate participants raised from their socio-economic levels (p-value = 0.001). Lice infestation reached higher rates in children aged less than 20 years with itching of the hair scalp. The obtained results revealed that 59.33% of the respondents believed that frequency of personal hygiene and washing of hair were the best methods for preventing the lice infestation. However, the treatment of lice infestation using anti-lice agents (p-value = 0.020) was preferred by 14.26% of participants. Although knowledge about the preventive tools for lice infestation (p-value 0.089) was not significantly associated with the experience of infestation but knowledge about the appropriate treatment to kill lice (p-value 0.020) and the wrong practices in the treatment of a head lice infestation were (p-value 0.005) significantly associated with the experience of infestation. Health programs and campaigns preventions are highly advised to increase the awareness of Pediculosis capitis with an effective strategic plan to control, manage, and prevent this disease.

Morphomolecular and pathological study of Scaphanocephalus sp. in new host Siganus argenteus in the Arabian Gulf.

In this study, Scaphanocephalus parasite (Platyhelminthes: Heterophyidae) metacercariae were found in Siganus argenteus (forktail rabbitfish or streamlined spinefoot) in the Arabian Gulf of Jubail province, Saudi Arabia. The findings may constitute new host and locality records for this parasite. Based on the number of black spots containing parasite cysts per fish, our study indicated that Siganus argenteus had high infection intensities of encysted metacercariae belonging to the genus Scaphanocephalus. Of the 3500 S. argenteus specimens examined, 800 (22.9%) showed multiple black cysts over the entire body surface, including the membranous parts of fins, while none were seen on the internal organs. The prevalence of infection was highest in summer (June-August) (8.8%). The excysted metacercariae differed morphologically from previously identified Scaphanocephalus spp. Molecular analysis of rDNA showed 100% identity with an unnamed Scaphanocephalus sp. reported in Caribbean fish. Therefore, our findings may indicate a new species of this previously rarely recorded fish parasite. The histopathological examination revealed that the encysted parasites were restricted to the dermal layer of the skin and surrounded by melanophores and a fibrous connective tissue capsule, with focal myositis and Zenker's necrosis in the underlying muscle tissue. The characteristically 'winged' parasite was clearly observed within the cysts. The high prevalence of Scaphanocephalus infection in siganid fish we detected requires further epidemiological study.

Prevalence Rate and Molecular Characteristics of Oestrus ovis L. (Diptera, Oestridae) in Sheep and Goats from Riyadh, Saudi Arabia.

Heads of sheep (n = 600) and goats (n = 800) slaughtered at Al-Aziziah Abattoir in Riyadh, Saudi Arabia, were inspected for the presence of O. ovis larvae (L). Heads were split along the longitudinal axes, and larvae (L1, L2, and L3) were gathered. The infestation rate was significantly higher in goats (44.5%; 356/800) than that in sheep (22.3%; 134/600). Out of the 151 collected larvae from sheep, 0% were L1, 1.3% were L2, and 98.7% were L3. Out of the total of 468 larvae from goats, 0% were L1, 1.2% were L2, and 98.8% were L3. The infestation rate was significantly higher in males than that in females. Myiasis-causing larvae collected from Riyadh, Saudi Arabia, were authenticated as O. ovis, according to morphological characteristics. Polymerase chain reaction (PCR) amplification of a partial fragment (600 bp) of the mitochondrial cytochrome c oxidase subunit I (mtCOI) gene further confirmed the species. Phylogenetic analysis based on the partial mtCOI gene sequence demonstrated that 23 unique sequences showed high similarity based on nucleotide pairs of O. ovis accessions retrieved from GenBank.

Anti-coccidial effect of Commiphora molmol in the domestic rabbit (Oryctolagus cuniculus domesticus L.).

This study investigated the effect of oleo-gum-resin of Commiphora monlmol Engler (Family: Burceraceae) known as Myrrh and the commercial extract known as Mirazid as a treatment against hepatic coccidiosis induced by the parasite Eimeria stiedae in domestic rabbits. Rabbits were infected with 1000 parasite sporulated oocysts and subjected to two treatment regimens, using crude myrrh suspension and the oleo-resin extract, mirazid, each administered at 500 mg/kg rabbit body weight. Treatments of infected rabbits resulted in significant reduction of the mean oocyst numbers in rabbit faeces by 52.38% in the crude-treated rabbits and by 90.90% mirazid-treated rabbits, compared to the untreated infected rabbits at day 21 post-infection (pi). At day 28 pi no oocysts parasite were observed in the faeces of rabbits. Both treatments resulted in significant recovery of infected rabbits from all symptoms of infection compared to the untreated infected group and healthy control groups. Histopathological examination of liver showed remarkable improvement in all histopathological parameters in G5 and G8 compared with the infected untreated G2. These included an almost complete healing of the hemorrhagic tissue and partial healing of the endothelial lining and hepatocytes encircling the central vein, the hepatocytes laminate regained their original radial shape and disappearance of fat vacuoles from the tissue and remarkable reduction in lymphocytes infiltration, decreased hyperplasia of the epithelial cells with significant decreasing of the parasite stage numbers. Results also indicate that mirazid was more effective than crude myrrh, probably due to higher content of purified active ingredients.