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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.

Strongylus vulgaris , a devastating parasitic nematode in equids, causes life-threatening verminous aneurysms that are challenging to diagnose early. This study pioneered integrating nanotechnology into an indirect enzyme-linked immunosorbent assay (i-ELISA) system to enhance the sensitivity and specificity for detecting S. vulgaris larval antigens in equine serum samples, with PCR confirmation of the species. A conventional i-ELISA and an innovative nano-based ELISA were developed using excretory-secretory antigens from adult S. vulgaris worms. The nano-ELISA incorporated gold nanoparticles (17.4–41.4 nm) conjugated with detection antibodies, enabling remarkable signal amplification. Of the 120 examined equines, 100 (83.33%) were positive for S. vulgaris infection. A conventional i-ELISA and an innovative nano-ELISA incorporating 17.4–41.4 nm gold nanoparticles were optimized using S. vulgaris excretory-secretory antigens. Both assays demonstrated high specificity, with no cross-reactivity against sera from animals infected with other helminth parasites. Remarkably, optical density (OD) readings from both i-ELISAs exhibited a positive quantitative correlation with infection intensity. The i-ELISA OD ranged from 0.45–0.74 (G3), 0.75–0.94 (G2), to 0.95–2.5 (G1). The nano-ELISA showed enhanced signal amplification, with OD ranging from 0.40–0.84 (G3), 0.85–0.99 (G2), to 1.0–3.5 (G1). This nanotechnology-amplified ELISA opens new, highly sensitive, and specific techniques for parasitic diagnosis in equine medicine. Its superior performance, facilitated by signal-amplifying gold nanoparticles, illuminates nanotechnology's potential in revolutionizing parasitological diagnostics for enhanced animal health and welfare management.

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 4 th 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 .

Purpose This study aimed to green synthesized copper nanoparticles (CNP) using Lupinus arcticus extract and assess the protoscolicidal effects and the effectiveness of these nanoparticles in experimental animal settings to combat hydatid cysts. Methods The protoscolicidal properties of CNP were examined in vitro on hydatid cyst PS via the eosin exclusion assay. The in vivo effectiveness of CNP at doses of 20, 40, and 80 mg/kg/day for 28 days on hydatid cyst-infected mice was determined by assessing the number, size, weight of hydatid cysts the gene expression levels of caspase-3, antioxidant (glutathione peroxidase (GPx) and superoxide dismutase (SOD)) and inflammatory cytokines (Interleukin-4 (IL-4), IL-10, and inducible nitric oxide synthase (iNOS)) in the infected mice were also assessed by Real-time PCE. Results Analyses revealed that the CNP had a spherical shape, ranging in size from 10 to 85 nm. The half-maximal inhibitory concentration (IC50) values for CNP against protoscoleces ranged from 39.9 to 341.3 µg/mL. Caspase-3 gene expression in the PS treated with CNP at 1/3 IC50, ½ IC50, and IC50 was 1.79-, 3.11-, and 5.39-fold change ( p  < 0.001). Upon administration of CNP, particularly at doses of 80 mg/kg, a significant decrease ( p  < 0.001) in hydatid cysts’ quantity, size, and weight was evident. CNP treatment caused a notable reduction in oxidative stress markers and inflammatory cytokines and increased the gene expression of antioxidant enzymes ( p  < 0.001), whereas it modulated the serum levels of liver function parameters. Conclusion The study indicated the potential of green-synthesized CNP for eliminating the protoscoleces and in managing hydatid cysts through leveraging its antioxidant and anti-inflammatory characteristics. Nevertheless, additional investigations are necessary to elucidate the precise mechanisms of action and evaluate its effectiveness in clinical trials. Clinical Trial Number Not applicable.

This study investigated the prevalence, morphology, molecular identification, and histopathological effects of larval tapeworms (plerocercoids) infecting the skeletal muscles of the Indian halibut ( Psettodes erumei ) collected from the coastal waters of the Arabian Gulf. Numerous oval or round blastocysts, measuring 13–26 mm, were found embedded within the muscular tissues of the Indian halibut, rendering the fish unsuitable for human consumption. Morphological and molecular analyses identified the plerocercoids as Dasyrhynchus giganteus (family Dasyrhynchidae), with an overall prevalence of 15.4%. The seasonal prevalence was the highest in summer (14.6%), followed by spring (10.6%), winter (4.4%), and autumn (3.5%). Infection rates increased with fish size. Histopathological examination revealed fibrous connective tissue capsules surrounding the larvae, causing muscular atrophy and degenerative changes, with few inflammatory eosinophilic cells. Molecular and phylogenetic analysis of the 28S rDNA gene sequences confirmed the specimens as D. giganteus , clustered closely with other sequences of D. giganteus with 100% bootstrap values. This study provided valuable insights into the parasitic infection dynamics, seasonal variation, molecular identification, and histopathological effects, highlighting the importance of monitoring fish for food safety and public health implications.

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.

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. Graphical Abstract

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.

The intestine is considered a habitat for both bacteria and parasites. In this study, many fecal bacterial isolates and the protozoan Blastocystis sp. were recovered from stool samples of individuals with gastrointestinal conditions. Isolated bacteria were tested for extracellular protease production, and the most potent producer was identified by 16SrDNA gene sequencing as P. aeruginosa FZM498. The enzyme was extracted and purified to electrophoretic homogeneity by the DEAE-Sepharose ion-exchanger and SDS-PAGE revealed a major band at 42.15 KDa. It exhibited maximal activity at 35 °C with thermostability at 60 °C (T1/2 = 200.04 min). It was most active at pH 8.0 and stable at 5.0–9.5. Enzymatic activity was greatly stimulated in the presence of Fe2+ ions, but was repressed by Zn2+ and Hg2+ ions. Inhibition by PMSF, TLCK, aprotinin, benzamidine, and SBTI protease reagents suggests a serine protease family. The Vmax and Km dynamic constants against azocasein were 36.232 U/mL and 0.0072 mM, respectively. It exhibited the lowest Km value against the synthetic substrate D-Val-Leu-Lys-pNA among all substrates, indicating a plasmin-like activity. Interestingly, when tested against Blastocystis sp., cysts appeared progressively shrunken, ruptured, and mycelial-like, indicating complete structural collapse with leakage of intracellular contents. The importance of this research is that it is the first study to test the anti-Blastocystis activity of an extracted bacterial serine protease from the gut. This could be a promising, eco-friendly, natural alternative as an anti-Blastocystis agent. The objective of this study was to isolate, purify, and biochemically characterize an extracellular serine protease produced by gut-associated bacteria, as well as to assess its in vitro anti-Blastocystis efficacy as a potential natural and ecologically friendly antiparasitic therapy.