Combating the causative agent of amoebic keratitis, Acanthamoeba castellanii, using Padina pavonica alcoholic extract: toxicokinetic and molecular docking approaches.

Natural products play a significant role in providing the current demand as antiparasitic agents, which offer an attractive approach for the discovery of novel drugs. The present study aimed to evaluate in vitro the potential impact of seaweed Padina pavonica (P. pavonica) extract in combating Acanthamoeba castellanii (A. castellanii). The phytochemical constituents of the extract were characterized by Gas chromatography-mass spectrometry. Six concentrations of the algal extract were used to evaluate its antiprotozoal activity at various incubation periods. Our results showed that the extract has significant inhibition against trophozoites and cysts viability, with complete inhibition at the high concentrations. The IC50 of P. pavonica extract was 4.56 and 4.89 µg/mL for trophozoites and cysts, respectively, at 24 h. Morphological alterations of A. castellanii trophozoites/cysts treated with the extract were assessed using inverted and scanning electron microscopes and showed severe damage features upon treatment with the extract at different concentrations. Molecular Docking of extracted compounds against Acanthamoeba cytochrome P450 monooxygenase (AcCYP51) was performed using Autodock vina1.5.6. A pharmacokinetic study using SwissADME was also conducted to investigate the potentiality of the identified bioactive compounds from Padina extract to be orally active drug candidates. In conclusion, this study highlights the in vitro amoebicidal activity of P. pavonica extract against A. castellanii adults and cysts and suggests potential AcCYP51 inhibition.

Genetic relatedness and diversity of Capillaria species infecting bayad (Bagrus bajad) in upper Egypt.

BACKGROUND: This study investigates the genetic characteristics of Capillaria isolates from the infected fish, Bagrus bajad, and their relation to human Capillaria philippinensis using Random Amplified Polymorphic DNA (RAPD-PCR) analysis. Fifteen fish Capillaria were isolated and compared to identified human C. philippinensis using six primers: M-are, M-1, G-7, G-11, G-15, and G-18. RESULTS: All six primers successfully amplified DNA, highlighting their efficacy in distinguishing between human and fish Capillaria isolates. The analysis revealed distinctive banding patterns between fish and human isolates, with variations in size and number of DNA fragments. Additionally, genetic similarity analysis showed intriguing patterns of relatedness, with certain pairs exhibiting high similarity percentages. Comparative assessment of RAPD polymorphism demonstrated consistent findings of 100% polymorphism across all primers. The Unweighted Pair Group Method with Arithmetic Mean Algorithm (UPGMA) evaluated the closest relationship between human and fish isolates. These results underscore the utility of RAPD analysis in delineating the genetic diversity among Capillaria isolates from different hosts. CONCLUSION: Overall, this study contributes to our understanding of the genetic variability and relatedness among Capillaria isolates, shedding light on their evolutionary dynamics and zoonotic potential.

New host record, Sclerophrys regularis (Bufonidae), for Rhabdias africanus (Rhabdiasidae, Kuzmin, 2001) based on molecular and morphological evidence.

With diversity of hosts range, most identified nematode species still lack the crucial connection between morphological and molecular make-up, which is important for precisely classifying the specimens. The present study provides the complete description of Rhabdias africanus in the new host record, Sclerophrys regularis. Fifty toad specimens were collected, and a high prevalence of R. africanus infection (74%) was observed. Morphology and ultrastructure were observed using light and scanning electron microscopes. Morphological characteristics, including peculiarities of the head, the shape and position of the lips, and the number of labial papillae, were described. The length of the body, the esophageal length, the distance from an anterior end to the nerve ring, and the tail length were reduced in the studied samples relative to previously described specimens. Furthermore, some variable matrices that have not previously been described, e.g., ovarian part widening, the nerve ring and its location, and eggs with different stages of larvae, were included in the present study. Genus and species identification was confirmed by comparing partial 12S (619 bp) and ITS (878 bp) gene sequences to those of Rhabdias species deposited in GenBank. The studied species showed a 99.34% resemblance to R. africanus from South Africa. We assume our findings will aid in the molecular identification of adult and larval stages of this genus in amphibians. We strongly recommend further studies on the environmental factors that promote Rhabdias infection and transmission.

Contact lenses contamination by Acanthamoeba spp. in Upper Egypt.

BACKGROUND: Acanthamoeba spp. are one of the free-living amoeba that spread worldwide causing keratitis. Owing to the increase in the use of lenses, whether for medical or cosmetic purposes, the incidence of disease increases every year. Contamination of the lenses with the Acanthamoeba trophozoites or cysts may lead to eye infection and cause sight-threatening keratitis in human. We isolated Acanthamoeba spp. from new lenses, used lenses, and contact lens disinfecting solutions and identified them based on morphological characteristics and molecular test. METHODS: New and used lenses and contact lens disinfecting solutions were cultured on monogenic media. Light and scanning electron microscope was used to identify Acanthamoeba spp. morphological features. Genotype identification was also evaluated using PCR sequencing of 18S rRNA gene specific primer pair JDP1 and JDP2. RESULTS: A hundred samples were examined, 29 (29%) were infected with Acanthamoeba spp. That belonged to two strains of Acanthamoeba (Acanthamoeba 41 and Acanthamoeba 68). 18S rRNA of the Acanthamoeba 41 had 99.69% sequence identity to Acanthamoeba castellanii clone HDU-JUMS-2, whereas Acanthamoeba 68 had 99.74% similar pattern to that of Acanthamoeba sp. isolate T4 clone ac2t4 that are morphologically identified as Acanthamoeba polyphaga. The obtained data revealed that the isolated strains belong to T4 genotype that was evolutionarily similar to strains isolated in Iran. CONCLUSIONS: Cosmetic lenses and disinfectant solutions are a major transmissible mode for infection. This genotype is common as the cause of Acanthamoeba keratitis. To avoid infection, care must be taken to clean the lenses and their preservative solutions and prevent contamination with the parasite.

Endocrine, Stemness, Proliferative, and Proteolytic Properties of Alarm Cells in Ruby-Red-Fin Shark (Rainbow Shark), Epalzeorhynchos frenatum (Teleostei: Cyprinidae).

The current study investigated the morphological, histochemical, and immunohistochemical characteristics of alarm cells and their precursors in ruby-red-fin shark (rainbow shark), Epalzeorhynchos frenatum (Teleostei: Cyprinidae). Precursor alarm cells were shown to be small, cuboidal, pyramidal, or round in shape, with eosinophilic cytoplasm, resting on the basement membrane of the epidermis. The cells later elongated to become columnar in shape. Subsequently, they enlarged and became large oval-shaped cells. They then underwent shrinkage and vacuolation. The superficial alarm cells were collapsed. Alarm cells were found to have an affinity for different histochemical stains, including bromophenol blue, iron hematoxylin, Sudan black, Mallory triple trichrome, Crossman's trichrome, Safranin O, and Weigert's stains, as well as lipase and alkaline phosphatase. Endocrine properties of the alarm cells were identified by silver staining and synaptophysin immunostaining. Alarm cells exhibited stemness activities and had a strong immunoaffinity for CD117. They had a proteolytic function, as identified by lysosome-specific staining with acridine orange and strong immunoaffinity for matrix metalloproteinase (MMP-9). They also exhibited proliferatively, reflected by immunological staining by proliferating cell nuclear antigen. In conclusion, alarm cells are unique epidermal cells with multiple functions. They play immunological, and endocrine, roles. They also retain stemness and proliferative properties.

An Insight into the Potential Parasitological Effect of Schistosoma mansoni Antigens in Infected Mice: Prophylactic Role of Cercarial Antigen.

Schistosomiasis is a multifactorial disease that includes environmental, behavioral, parasitic, vector, and host factors. This study aimed to assess the protective effect of single and polyvalent antigens from cercarial antigen preparations (CAPs), soluble worm antigen preparations (SWAPs), and soluble egg antigens (SEAs) which were used as candidate vaccines in an experimental model of Schistosoma mansoni-infected mice. The efficiency of the antigens was tested by determining their effects on fecal egg count, egg viability analysis, and tissue egg counts. Histological and morphometric analyses of granulomas in liver and intestine tissues were performed. In the present study, all immunized groups showed a significant reduction in the average fecal egg count and tissue egg load compared with infected mice. The most substantial reduction in fecal egg count was observed in the combined vaccinated group (23.23 ± 3.2). The group vaccinated with CAP before infection showed the highest reduction in tissue egg load (liver and intestine: 85.22 and 91.70%, respectively). Immunized animals showed a highly significant reduction in the numbers of hepatic granulomas compared with the infected non-immunized group. In conclusion, combining these different antigens (CAP, SWAP, and SEA) augments the protective immunity compared with other immunized groups.

Evaluation and Microanalysis of Parasitic and Bacterial Agents of Egyptian Fresh Sushi, Salmo salar.

The present study aimed to evaluate the quality of fresh sushi in Egypt. Fifty samples of sushi (Salmo salar) were collected from restaurants in Alexandria, Egypt. Paraffin, semi-thin and ultra-thin sections were used for parasitological analysis by light and transmission electron microscopy. Bacteria were isolated by the dilution plate and direct plate methods and identified by a Vitek system. Twenty (40%) of the total examined samples showed microsporidia and helminth metacercariae infections. Histochemical stains showed distinct pinkish-red pyriform microspores embedded in muscular tissue stained with Gram, periodic acid-Schiff (PAS), and Ziehl-Neelsen (ZN) stains. Semi-thin sections showed double membrane xenoma-inducing granulomas containing spores at different developmental stages. Empty sporophorous vesicles and free spores were observed in the electron microscopic images. A bacteriological assay showed forty samples (80%) contaminated with human pathogenic bacteria with the average total bacterial counts ranging from 32 to 526 CFU/g. Four species of human pathogenic bacteria were identified in the examined samples, namely Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, and Serratia plymuthica in 40, 38, 11, and 6 samples, respectively. These constitute the first record of fresh sushi product in Egypt and indicate the potential pathogenicity associated with raw seafood products.

Protozoan and helminthes parasites endorsed by imported camels (Camel dromedaries) to Egypt.

The prevalence and species spectrum of some blood and intestinal parasites affecting imported camels was studied on a total of 120 clinically suspected camels (males) imported to Egypt from Sudan during the period from January till July 2016 in Abu-Simbel quarantine station, Aswan governorate. Blood and fecal samples were collected from all camels under the study. The fecal samples were collected and examined by sedimentation-floatation techniques for detection of parasitic eggs/oocysts. Coprological examination revealed that the prevalence rate of the parasitic infection was 60% (72 out of 120). Eighteen species of helminthes/protozoan parasites eggs/oocysts were encountered stongyles species were the hightest prevalent of nematodes 12.5%. Four genera of flat worms were identified in the present study including Paramphistomum sp. 0.8%, Fasciola sp. 3.3%, Moniezia sp. 7.5% and Dicrocoelim sp. 0.8%. Four species of Eimeria were identified (E. cameli, E. dromedarii, E. rajasthaniand E. pellerdyi) in infected camels the commenst one is E. cameli 15.8%, Cryptospridium sp. and Balatidium coli were recorded with a prevalence rate about 15.8%, 8.3% and 6.7% respectively. Blood smears from jugular vein revealed that 2.5% of camels were infected with Dipetalonema evansi. Wide spectrum and high prevalence of internal parasites were observed in the present study which may be lead to severe economic losses, so the application of control measures and treatment of infected camels with specific and effective drugs during the quarantine period are most important to prevent spreading of parasitic infestation and/or introduction of parasites previously not exist in our country.

The protective role of quince leaf extract against the adverse impacts of ultraviolet--a radiation on some tissues of Clarias gariepinus (Burchell, 1822).

In the present study the protective role of quince leaf extract against the adverse impacts of ultraviolet radiation-A (UVA) on some tissues of Clarias gariepinus (Burchell, 1822) was considered. Fishes were classified into four groups: control, UVR-treated group (for 3days/for 3h/day), UVR-treated group (for 3days/for 3h/day) with adding 10ml of quince extract, and UVR-treated group (for 3days/for 3h/day) with adding 20ml of quince leaf extract. Blood smears and sections of the liver, and skin were processed routinely for H & E paraffin embedding technique. Some UVA-induced malformations were recorded in the red blood cells including crenated cells (Cr), Acanthocytes (Ac), tear drop-like cells (Tr) and sickle cells (Sk). Also, UVA-induced disorganization of the normal architecture of hepatic tissues with lipidosis was evident. Hypertrophy and vacuolated club cells were recorded in skin exposed to UVA. In conclusion, quince leaf extract has a valuable antioxidant protective role to prevent and/or repair the histopathological changes induced by UVA.