Parasitology, Animal Science and Zoology, Veterinary
30
Scopus Publications
Scopus Publications
Bridging biology and therapy: translational advances of extracellular vesicles in veterinary clinical practice Walaa A. Gad, Sally Ibrahim, Hiam Nagdy, Bassma S. M. Elsawy, Dina Aboelsoued, et al. Veterinary Research Communications, 2026 Extracellular vesicles (EVs) are small membrane-bound particles released by numerous cell types and are gaining popularity in veterinary medicine due to their extensive biological activities and therapeutic potential. This review summarizes the classification, biogenesis, and molecular cargo of various types of EVs, such as exosomes, microvesicles, and apoptotic bodies, as well as their emerging roles in cellular communication, diagnostics, and therapeutics across a wide range of veterinary applications. Beyond mesenchymal stem cell (MSC)-derived EVs, EVs from immune cells, pathogens, and body fluids show great promise for tissue healing, immunological regulation, infectious disease management, drug delivery systems, vaccine development, and reproductive health. We critically evaluate recent advancements, limitations, and future possibilities in using EVs to improve diagnosis and treatment results in veterinary species. The review’s goal is to provide a comprehensive picture of the rapidly increasing EV landscape and to make it easier to incorporate EV-based technology into clinical veterinary practice.
Coproantigen detection and molecular identification of Cryptosporidium species among newborn and adult farm animals Dina Aboelsoued, Nagwa I. Toaleb, Kadria N. Abdel Megeed AMB Express, 2025 Cryptosporidium sp. is an obligatory intracellular apicomplexan protozoan parasite that causes a disease called cryptosporidiosis with substantial veterinary and medical importance. Therefore, this study aimed to evaluate an early diagnosis of cryptosporidiosis using the anti-Cryptosporidium parvum oocyst immunoglobulin IgG polyclonal antibodies (anti-C. parvum IgG PAbs)-based sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Cryptosporidium oocyst antigens in fecal samples of farm animals in Egypt. Further molecular identification and sequencing were performed for the detected isolates. Eight hundred and twenty fecal samples of farm animals; 102 buffalo calves, 120 cattle calves, 100 lambs and 98 goat kids, 80 buffaloes, 60 cattle, 160 sheep and 100 goats, collected from different small-scale farms and local holders were examined for cryptosporidiosis by Modified Ziehl-Neelsen (MZN) technique. The percentage of positivity was 45.1%, 50%, 20%, 18.4%, 31.25%, 38.3%, 18.8%, and 11% in buffalo calves, cattle calves, lambs, goat kids, adult buffaloes, adult cattle, sheep, and goats, respectively. Molecular identification of Cryptosporidium samples was performed based on COWP gene, revealing the isolates: GenBank: OQ121955.1, OR029973.1 and PP316107.1 which were identical to the C. parvum and GenBank: PP316108.1 and OR029972.1 which were identical to C. hominis and C. andersoni, respectively. Then, C. parvum oocysts were used for preparation of antigens and rabbit immunization. Anti-C. parvum IgG PAbs were purified and characterized by SDS-PAGE and then labeled with horseradish peroxidase (HRP). Anti-C. parvum IgG PAbs in-house sandwich ELISA was prepared, then tested this ELISA on 820 samples and compared results with MZN microscopical examination and a commercial sandwich ELISA kit. In this study, in-house sandwich ELISA scored higher sensitivity of 98%, 100% specificity, validity 99% and relative agreement 98.6% than (92%, 90%, 91% and 91.4%) of MZN and (96%, 95%, 95.5% and 95.7%) of coproantigen commercial sandwich ELISA kit, respectively. Moreover, we used PCR to evaluate the positivity of in-house sandwich ELISA results, and the total PCR positive samples were 263 out of 268 sandwich ELISA positive samples (98.13%). In conclusion, the prepared Anti-C. parvum IgG PAbs based sandwich ELISA offered a simple and accurate diagnostic method for cryptosporidiosis in the fecal samples of different species of farm animals in Egypt with high sensitivity (98%) and specificity (100%). Further studies on this Anti-C. parvum IgG PAbs may help also in the protection against cryptosporidiosis.
Coprological and molecular prevalence of Cryptosporidium and Giardia in cattle and irrigation water from Beni-Suef Governorate, Egypt Fatma El-zahraa Ramadan Saleh, Hend H. A. M. Abdullah, Dina Aboelsoued Scientific Reports, 2025 Cryptosporidium and Giardia are globally significant protozoan parasites responsible for severe foodborne and waterborne outbreaks, posing substantial zoonotic and environmental risks. This study aimed to comprehensively assess the prevalence of cryptosporidiosis, giardiasis, and co-infections in Beni-Suef Governorate, Egypt, using an integrated diagnostic approach combining microscopy and molecular techniques. Additionally, it was sought to identify associated risk factors in cattle fecal samples. Microscopical examination of 970 cattle fecal samples revealed an overall infection rate of 67.42% (654/970), with Cryptosporidium detected in 42.68% (414/970), Giardia in 11.96% (116/970), and co-infections in 12.78% (124/970) of cases. In irrigation water, Cryptosporidium oocysts and Giardia cysts were detected in 2/24 (8.33%) and 1/24 (4.16%) of samples, respectively. Molecular and phylogenetic analyses identified Cryptosporidium hominis in cattle and, for the first time in Egypt, Cryptosporidium ubiquitum and Cryptosporidium ryanae in irrigation water, while also proving the presence of Cryptosporidium bovis and Giardia assemblage A in cattle. Risk factors, including sex, age, season, and fecal consistency, significantly influenced infection rates, with higher prevalence in females, calves under two months, spring season, and diarrheic feces. These findings underscore the urgent need for One Health-based control strategies, integrating targeted interventions to mitigate the burden of Cryptosporidium and Giardia infections and environmental contamination.
Sarcocystis species: molecular identification and seroprevalence in water buffaloes (Bubalus bubalis) Nagwa I. Toaleb, Raafat M. Shaapan, Haitham Elaadli, Kadria N. Abdel Megeed, Dina Aboelsoued BMC Veterinary Research, 2025 Meat infection with the coccidian protozoan zoonotic Sarcocystis spp. causes public health hazards and high economic loss. The current study aimed to investigate molecular identification, cyst histological examination and seroprevalence of Sarcocystis spp. in water buffaloes slaughtered in the main abattoirs of 6 Egyptian Governorates; Cairo, Giza, Beni-suef, Al-Sharqia, Qalyubia, and El-Beheira. Each buffalo was visually inspected for the presence of Sarcocystis macrocysts and blood samples were collected. Out of 900 examined water buffaloes (Bubalus bubalis), 246 (27.3%) were found to be infected based on macroscopic examination. Histological examination of the macrocyst revealed metrocytes close to the cyst's edge and elongated curving basophilic bradyzoites occupying most of the cyst. Depending on age, we categorized naturally infected buffaloes into 3 groups: young (< 2 y.), adult (2–5 y.) and old (5–10 y.). The highest infection rate was observed in older buffaloes aged 5–10 years, meanwhile female animals exhibited a higher prevalence of infection compared to males. The esophagus had the highest presence of Sarcocystis compared to other organs. Using PCR based on 18S rRNA gene and sequencing, we isolated 3 Sarcocystis species from infected tissues: S. buffalonis, S. fusiformis and S. hirsuta-like. The prepared whole cyst antigen of S. fusiformis was characterized by 9 major polypeptides (140, 120, 78, 66, 53, 39, 32, 24 and 19 kDa) on 10% SDS-PAGE. Using western blotting, we identified 2 common immunogenic reactive bands (66 and 32 kDa) against naturally infected buffalo sera and hyperimmune rat sera. Additionally, sarcocystosis positive seroprevalence rate was 46.4% (418/900) with a sensitivity of 97.6% and specificity of 100% using indirect ELISA based on S. fusiformis whole cyst antigen. In conclusion, this study highlights the molecular identification, cyst histological examination and seroprevalence of Sarcocystis sp. among buffaloes in Egypt. ELISA using the S. fusiformis whole cyst antigen could be adapted to detect antibodies to Sarcocystis sp., in live animals, with an acceptable specificity and sensitivity.
Comparison of a commercial ELISA, an in-house indirect ELISA, and a dot-ELISA developed for the serodetection of Toxoplasma gondii antibodies in farm animals Nagwa I. Toaleb, Raafat M. Shaapan, Dina Aboelsoued Journal of Veterinary Diagnostic Investigation, 2025 Toxoplasma gondii is a widespread intracellular protozoan that can infect humans and animals. We isolated T. gondii strains from sheep, goats, cattle, buffaloes, and camels to develop and evaluate a modified in-house dot-ELISA for the detection of Toxoplasma antibodies in farm animals, and compared the results with a commercial ELISA (IDvet; gold standard). Animal tissue samples ( n = 430) were examined microscopically, and infected tissues were bioassayed in mice as a viability test. Egyptian Toxoplasma strains were isolated from sheep, cattle, and camels and identified via PCR using the B1 gene (GenBank OR837022.1, OR837021.1, OR837020.1 from sheep, cattle, and camels, respectively). A T. gondii tachyzoite antigen from a sheep strain had the highest potential for the detection of specific T. gondii antibodies. We characterized this antigen using SDS-PAGE and separated it into 10 polypeptides of 96−12 kDa. Our modified in-house dot-ELISA detected T. gondii seropositivity in 172 of 430 (40%) farm animals with a sensitivity of 96.6% and specificity of 100%. The results of our dot-ELISA were confirmed in comparison with those of our indirect ELISA and the commercial ELISA. In a western blot, a predominant immunogenic reactive antigen band of 65 kDa was detected in T. gondii –positive sera of sheep, cattle, buffaloes, and camels; no cross-reaction occurred with antibodies to other parasitic infections or samples from healthy controls. Our modified in-house dot-ELISA is a rapid and simple test that showed promise for the detection of Toxoplasma antibodies in farm animals.
Anticryptosporidial action mechanisms of Launaea spinosa extracts in Cryptosporidium parvum experimentally infected mice in relation to its UHPLC-MS metabolite profile and biochemometric tools Mai M. Elghonemy, Mohamed G. Sharaf El-Din, Dina Aboelsoued, Mohamed F. Abdelhameed, Mohamed A. El-Saied, et al. Plos One, 2025 Background Cryptosporidium parvum, a leading cause of diarrhea, is responsible for millions of food and waterborne illnesses in humans and animals worldwide. Launaea spinosa (Asteraceae family) is a common herb found in the desert of the Mediterranean region, encompassing the peninsula of Sinai. Traditionally, it has been utilized for managing gastrointestinal issues and inflammation. Methods and findings The present study aimed to assess Launaea spinosa (LS) extracts viz. ethyl acetate (LS-EtOAc), ethanol (LS-EtOH), and n-butanol (LS-BuOH), of different polarities against C. parvum in experimentally infected mice based on immunological, biochemical, histo- and immunohistochemical assays. Extracts were characterized via UHPLC-ESI-LIT-Orbitrap-MS and metabolite profiles were subjected to correlation modeling with bioactivities via supervised Partial Least Square (PLS) to identify active agents. Most L. spinosa extracts reduced fecal C. parvum oocyst count and mucosal burden (P < 0.05) than untreated infected mice, with LS-BuOH (200 mg/kg) exerting the highest reduction percentage (97%). These extracts increased immunoglobulin G (IgG) levels in infected and treated mice at all examined days post treatment. Also, the highest Interferon-Gamma (IFN-γ) and Interleukin-15 (IL-15) levels were obtained after 10 days of post inoculation (dPI), which were restored to a healthy state after 21 days, concurrent with a decrease in Tumor Necrosis Factor-Alpha (TNF-α) (P < 0.001). The increased liver enzyme (alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase) levels with infection were likewise reduced with extract administration. The LS extracts caused a significant increase in antioxidant glutathione peroxidase (GSH-Px) and catalase (P < 0.001). Examination of colon tissue revealed that infected-treated mice with LS extracts exhibited a reduction in the expression of cleaved caspase-3, damage score, and degenerative changes. Metabolite profiling of different L. spinosa extracts led to the identification of 86 components, primarily phenolic acids, flavonoids, triterpenoid saponins, and fatty acids, with the first report of sulfated triterpenoid saponins in Launaea genus. PLS regression analysis revealed that bioeffects were significantly positioned close to LS-BuOH extract (R2: 0.9) mostly attributed to triterpenoid saponins and flavonoid glycosides. Conclusions This study demonstrated potential anti-cryptosporidial effects of LS extracts, especially LS-BuOH, suggesting its potential for inclusion in future nutraceuticals aimed at C. parvum treatment.
In vitro and ex vivo protoscolicidal effect of poly(amidoamine) nanoemulsion against Echinococcus granulosus Dina Aboelsoued, Nagwa I. Toaleb, Sally Ibrahim, Saber Ibrahim Scientific Reports, 2024 Hydatidosis causes a serious health hazard to humans and animals leading to significant economic and veterinary and public health concern worldwide. The present study aimed to evaluate the in vitro and ex vivo protoscolicidal effects of synthesized poly(amidoamine), PAMAM, nanoemulsion. In this study, PAMAM was characterized through dynamic light scattering technique to investigate the particle size and zeta potential of nanoemulsified polymer. For the in vitro and ex vivo assays, we used eosin dye exclusion test and scanning electron microscope (SEM) to evaluate the effects of the prepared and characterized PAMAM nanoemulsion against protoscoleces from Echinococcus granulosus sensu lato G6 (GenBank: OQ443068.1) isolated from livers of naturally infected camels. Various concentrations (0.5, 1, 1.5 and 2 mg/mL) of PAMAM nanoemulsion at different exposure times (5, 10, 20 and 30 min) were tested against protoscolices. Our findings showed that PAMAM nanoemulsion had considerable concentration- and time-dependent protoscolicidal effect at both in vitro and ex vivo experiments. Regarding in vitro assay, PAMAM nanoemulsion had a potent protoscolicidal effect when compared with the control group with a highest protoscolicidal activity observed at the concentration of 2 mg/mL at all exposure times, such that 100% of protoscolices were killed after 20 min of exposure. Also, the mortality of protoscolices was 100% after 30 min of exposure to 1 and 1.5 mg/mL of PAMAM nanoemulsion, in vitro. Concerning ex vivo assay PAMAM nanoemulsion recorded the highest mortality rates at the concentration of 2 mg/mL (55, 99.4 and 100% at 10, 20, 30 min, respectively). Ultrastructure examination of examined protoscolices after 20 min of exposure to PAMAM nanoemulsion showed a complete loss of rostellar hooks, disruption of suckers with disorganization of hooks with partial or complete loss of them, and damage of protoscolices tegument with loss of their integrity in the form of holes and contraction of the soma region were observed in 1.5 and 2 mg/mL of PAMAM, in vitro and ex vivo, showing more damage in the in vitro conditions. It can be concluded that PAMAM nanoemulsion is a promising protoscolicidal agent offering a high protoscolicidal effect at a short exposure time. Further in vivo studies and preclinical animal trials are required to evaluate its efficacy and clinical applications against hydatid cysts.
Serodiagnosis of nasal myasis in camels (Camelus dromedaries) in Egypt using third larval instar affinity-purified glycoprotein Dina Aboelsoued, Nagwa I. Toaleb, Amany M. Mohamed, Kadria N. Abdel Megeed, Sahar Hussein Abdalla Hekal Veterinary Research Communications, 2024 The larvae of Cephalopina titillator cause nasopharyngeal myiasis in camels, which parasitize the living tissues of the nasal and paranasal sinuses, pharynx, and larynx. C. titillator infestation adversely affects camel health, meat, and milk production, and can even cause death. In our study, to improve the immunodiagnosis of camel nasal myiasis, a sensitive and specific enzyme-linked immunosorbent assay (ELISA) was developed and evaluated using the Concanavalin-A (Con-A) affinity purification for the C. titillator-N-acetylglucosamine (Ct-GlucNAc) glycoprotein fraction from third larval instars as an antigen for detecting C. titillator antibodies. Crude antigens were prepared from larval instars of C. titillator and evaluated by indirect ELISA. The third C. titillator larval antigen (L3Ct) had the highest protein content (P < 0.001) and the best diagnostic value; chi-square = 235 (P < 0.001). Four glycoprotein fractions were purified separately from the L3Ct antigen by Con-A purification and evaluated. The Ct-GlucNAc glycoprotein fraction was the fraction of choice with the highest diagnostic accuracy (P < 0.05). Using Ct-GlucNAc as a coating antigen, indirect ELISA showed a 99.3% sensitivity for positive results in camel myiasis samples and 100% specificity for negative results in healthy camel samples. The diagnostic accuracy was 99.7%, and no cross reactivity was detected for other parasitic diseases. The indirect ELISA results were confirmed by the western immunoblotting which was characterized by comparing sera from naturally infested dromedary camels with C. titillator, sera from healthy camels and sera from camels with other parasitic infections (Echinococcus granulosus, Fasciola gigantica, Hard ticks; Hyalomma dromedarii, Trichostronglid sp., Eimeria spp., and Cryptosporidium sp.). Immunoreactive antigenic bands of 63, 50, 30 and 18 kDa were predominantly detected in sera from camels with nasopharyngeal myiasis and didn’t react with healthy and camel’s sera from other parasitic infections. However, seven immunoreactive bands appeared at 120, 70, 63, 48, 35, 29, and 19 kDa in the crude L3Ct antigen. In addition, a positive rate of C. titillator immunodiagnosis was detected by indirect ELISA (48.6%, chi-square = 483, P < 0.001), which was significantly greater than that of postmortem diagnosis (31%). In conclusion, the current study introduces a new diagnostic immunoaffinity glycoprotein fraction of C. titillator 3rd larval instar-based ELISA as a highly accurate, simple and fast method to detect specific antibodies of nasal myiasis in camels.
