High potency of linalool-zinc oxide nanocomposite as a new agent for cystic echinococcosis treatment.

This current in vitro, ex vivo, and in vivo research aims to evaluate and analyze the linalool-zinc oxide nanocomposite (Lin-ZNP) for treating cystic echinococcosis. Lin-ZNP was synthesized using an ethanolic solution of polyvinyl alcohol. The protoscolicidal effects of Lin-ZNP were tested on hydatid cyst protoscoleces (PTS) in both in vitro and ex vivo by eosin exclusion test. The study also examined the impact on caspase-3 gene expression and the external structure of PTS. The in vivo effect was measured by examining hydatid cysts' quantity, dimensions, and weight in mice intraperitoneally infected with 0.5 mL of PTS solution containing 1,000 PTS. The antioxidant and inflammatory cytokine gene expression levels were examined using real-time PCR. Lin-ZNP significantly (P < 0.001) killed the PTS in both in vitro and ex vivo in a dose- and time-dependent manner. The treated PTS exhibited creases and protrusions as a result of bleb formation and upregulation in the gene expression of caspase-3. Upon treatment with Lin-ZNP, there was a significant (P < 0.001) reduction in the number, diameter, and weight of the hydatid cysts. Treatment with Lin-ZNP nanocomposite led to a significant increase in the expression of antioxidant genes and a notable decrease in oxidative stress markers, and the expression levels of IL-4 and IL-10. Lin-ZNP has the potential to act as a scolicidal agent and demonstrates promise in controlling hydatid cysts in a mouse model, attributed to its antioxidant and anti-inflammatory properties. However, additional studies in clinical trials are needed to confirm the use of Lin-ZNP for treating hydatidosis.

Potential Anti-Toxoplasmosis Efficiency of Phoenix dactylifera Extracts Loaded on Selenium Nanoparticles.

Background: Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii that infects humans and many types of mammals and birds. Objective: To investigate the effect of selenium nanoparticles (SeNPs) and Phoenix dactylifera (Pd) extracts loaded on SeNPs as a new agent to combat chronic T. gondii infections in murine model as an alternative method to standard Spiramycin drug therapy. Methods: A total of 64 female mice were randomly divided into eight groups: GI: Normal control, GII: Positive control, GIII: infected and treated with Spiramycin, GIV: infected and treated with SeNPs, GV: infected and treated with aqueous extract of Pd, GVI: infected and treated with methanolic extract of Pd, GVII: infected and treated with aqueous extract of Pd loaded on SeNPs, GVIII: infected and treated with methanolic extract of Pd loaded on SeNPs. Date palm (P. dactylifera) fruits were identified and collected from the farms of Saudi Arabia. Preparation and characterization of SeNPs were done. The parasitological, histopathological examinations and biochemical changes were evaluated in all groups. Results: Parasitological results showed significant differences in GVII in comparison to GII while GVIII showed significant differences in comparison to GII and GIII. The histopathological section of the cerebral cortex showed obvious alterations in the infected compared with untreated control groups. Aqueous and methanolic extracts of P. dactylifera loaded on SeNPs treatment showed improvement that indicated by few perivascular cuffing with few inflammatory cell infiltrations. Few granule cells with mild intracellular vacuolation and edema few deformed neurons with deep pyknotic nuclei. Microglia cells expression of Iba-1 and inflammatory cytokines (IL-4, IL-10 and INF-γ) in serum of all groups was higher in GII and lowest in GVIII followed by GVII. Conclusion: SeNPs and P. dactylifera extracts loaded on SeNPs could be a potent agent to combat T. gondii infections.

Molecular Characterization of Fasciola hepatica in Sheep Based on DNA Sequences of Ribosomal ITS-1.

Introduction: World Health Organization (WHO) considers Fascioliasis as a neglected tropical disease that requires global efforts for disease control. Data from the genetic characterization of Fasciola population shed light on the spread of infections among animals which could help in the development of effective parasite control. The aim of the present work was to genetically characterize Fasciola adult worms isolated from sheep in Saudi Arabia by sequence analysis of ITS-1 region. Methods: A total of 12,653 slaughtered sheep in Jeddah city, Saudi Arabia were examined for the presence of Fasciola spp. adult worms. The ITS-1 region of all parasites was amplified and sequenced. Results: Overall, 12 variants DNA sequences were obtained. The variance of isolates ranged from 0.00771 to 0.34405. BLAST search showed that all obtained sequences were Fasciola hepatica and had >99.3% similarity with F. hepatica isolates from Spain and USA (from different hosts other than sheep). Phylogenetic analysis showed that Fasciola isolates were closely related to isolates from different countries. Discussion: The current study showed that F. hepatica was the only spp. isolated from sheep in Jeddah. Further studies from different localities in Saudi Arabia are needed to help in the development of disease control.

Supplementing broiler diets with bacterial selenium nanoparticles enhancing performance, carcass traits, blood indices, antioxidant status, and caecal microbiota of Eimeria tenella-infected broiler chickens.

Nanomedicine is a critical therapeutic approach for treating most poultry illnesses, particularly parasitic infections. Coccidiosis is a severe protozoan infection affecting poultry; the emergence of drug-resistant Eimeria strains demands the development of new, safe therapies. Consequently, the objective of this work was to investigate the efficacy of the biosynthesized selenium nanoparticles (SeNPs) by Paenibacillus polymyxa (P. polymyxa) against Eimeria tenella (E. tenella) experimental infection in broiler chickens. The prepared SeNPs absorbed the UV at 270 nm were spherical with a size of 26 nm, and had a surface negative charge of -25 mV. One hundred and fifty, 1-day-old male broiler chicks were randomly allocated into 5 groups (30 birds/group with triplicates each) as follows: T1: negative control (noninfected and nontreated with SeNPs); T2: delivered SeNPs (500 µg/kg diet) for 35 successive days, T3: E. tenella-infected (positive control birds), T4: E. tenella-infected and treated with SeNPs (500 µg/kg diet) and T5: E. tenella-infected chicks and treated with anticoccidial agent (sulfadimidine, 16% solution 8 mL/L of drinking water) for 5 successive days. At 14 d of age, each bird in infected groups was orally treated with 3 × 103 sporulated oocyst of E. tenella. SeNPs considerably decreased the number of oocysts in broiler feces compared to positive control and anticoccidial drug, followed by a substantial reduction of parasite phase count in the cecum (15, 10, and 8 for meronts, gamonts, and developing oocysts) when compared with positive control birds. The Eimeria experimental infection lowered the activity of antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx), and reduced glutathione (GSH) while increasing the stress parameters nitric oxide (NO) and malonaldehyde (MDA). Moreover, the production of proinflammatory (TNF-α and IL-6) and apoptotic genes (BcL2 and Cas-3) were significantly elevated. Administrating SeNPs to chicks significantly decreased oxidative stress, inflammation, and apoptotic markers in the cecum tissue. Therefore, growth performance, carcass weights, antioxidant enzymes, and blood properties of infected chicks were enhanced. The findings compared the protecting role of Se-nanoparticles against cecum damages in E. tenella-infected broilers.

Annona muricata Leaf as an Anti-Cryptosporidial Agent: An In Silico Molecular Docking Analysis and In Vivo Studies.

Cryptosporidiosis is a serious parasitic diarrheal disease linked to the occurrence of colorectal cancer in immunocompromised patients. The FDA-approved drug nitazoxanide (NTZ) achieved a temporary effect, and relapses occur. Annona muricata leaf is widely used in traditional medicine to treat a wide range of disorders, including antiparasitic and anticancer effects. So, this study aimed to investigate Annona muricata leaf antiparasitic and anticancer properties compared to NTZ in Cryptosporidium parvum (C. parvum) acutely and chronically infected immunosuppressed mice. A molecular docking analysis was performed to evaluate the effectiveness of some biologically active compounds that represented the pharmacological properties of Annona muricata leaf-rich extract toward C. parvum lactate dehydrogenase compared to NTZ. For the in vivo study, eighty immunosuppressed albino mice were classified into four groups as follows: group I: infected and treated with A. muricata; group II: infected and treated with nitazoxanide; group III: infected and received no treatment; and group IV: were neither infected nor treated. Furthermore, half of the mice in groups I and II received the drugs on the 10th day post-infection (dpi), and the other half received treatment on the 90th day post-infection. Parasitological, histopathological, and immunohistochemical evaluations were performed. The docking analysis showed that the lowest estimated free energy of binding of annonacin, casuarine, L-epigallocatechin, P-coumaric acid, and ellagic acid toward C. parvum LDH, were -6.11, -6.32, -7.51, -7.81, and -9.64 kcal/mol, respectively, while NTZ was -7.03 kcal/mol. Parasitological examination displayed a significantly high difference in C. parvum oocyst mean counts in groups I and II compared to group III (p-value < 0.001), with group I demonstrating the highest efficacy. The analyses of histopathological and immunohistochemical results revealed that group I showed restoration of the normal villous pattern without evidence of dysplasia or malignancy. A. muricata leaf has proved to be a reliable agent for Cryptosporidium treatment. This paper argues for its promising use as an antiparasitic agent and for the prevention of neoplastic sequels of Cryptosporidium infection.

Identification and Genetic Characterization of Fasciola hepatica Isolated from Cattle in Jeddah, Saudi Arabia Based on Sequence Analysis of Mitochondrial (COI) Gene.

Background: In Saudi Arabia, more than US$ 0.2 million annual losses are caused by liver condemnations due to fascioliasis. Data obtained from the genetic characterization of Fasciola population sheds light on parasite transmission which could eventually help in development of effective parasite control measures. So, the aim of this study was to investigate the genetic diversity of Fasciola spp. isolated from cattle in Saudi Arabia by sequence analyses of COI gene. Materials and Methods: A total of 325 cows slaughtered at the central municipal abattoir in Jeddah city, Jeddah Province, Saudi Arabia were examined for fascioliasis in the period from 1st of June to 1st of July 2020. DNA was extracted from adult Fasciola worms and used for PCR and DNA sequence using a primer pair targeting COI gene. Analysis of the obtained sequences was done using BLAST search and phylogenetic analysis. Results: Bovine fascioliasis was diagnosed in 18 out of 325 cattle (5.5%). Forty-eight flukes were extracted from infected animals and DNA was successfully amplified from all flukes. Overall 12 different DNA sequences were obtained. BLAST search showed that all obtained sequences were F. hepatica and had >97% similarity with F. hepatica isolates from Tanzania, Europe and Iran. Phylogenetic analysis of the obtained sequences showed that Fasciola isolates from the current study were clustered in one subclade closely related to isolates from North and South Africa and Italy. Conclusion: Reports on the molecular characterization of Fasciola spp. in Saudi Arabia are limited. In the current study, our findings showed that F. hepatica was the only Fasciola species parasitizing cattle in Jeddah city, Saudi Arabia. Further studies using a large number of samples from different localities in Saudi Arabia are needed to provide data that will help the development of control measures against fascioliasis.

Clinical Characteristics, Outcomes and Prognostic Factors for Critical Illness in Hospitalized COVID-19 Patients in Saudi Arabia: A Retrospective Cohort Study.

Background: A good understanding of the possible risk factors for coronavirus disease 19 (COVID-19) severity could help clinicians in identifying patients who need prioritized treatment to prevent disease progression and adverse outcome. In the present study, we aimed to correlate clinical and laboratory characteristics of hospitalized COVID-19 patients to disease outcome in Saudi Arabia. Materials and Methods: The present study included 199 COVID-19 patients admitted to King Fahd Specialist Hospital, Buraydah, Qassim, Saudi Arabia, from April to December 2020. Patients were followed-up until discharge either for recovery or death. Demographic data, clinical data and laboratory results were retrieved from electronic patient records. Results: Critical COVID-19 cases showed higher mean of age and higher prevalence of co-morbid conditions. Fifty-five patients died during the observation period. Risk factors for in hospital death for COVID 19 patients were leukocytosis (OR 1.89, 95% CI 1.008-3.548, p = 0.081), lymphocytopenia (OR 2.152, 95% CI 1.079-4.295, p = 0.020), neutrophilia (OR 1.839, 95% CI 0.951-3.55, p = 0.047), thrombocytopenia (OR 2.152, 95% CI 0.852-5.430, p = 0.085), liver injury (OR 2.689, 95% CI 1.373-4.944, p = 0.003), acute kidney injury (OR 1.248, 95% CI 0.631-2.467 p = 0.319), pancreatic injury (OR 1.973, 95% CI 0.939-4.144, p = 0.056) and high D dimer (OR 2.635, 95% CI 0.747-9.287, p = 0.091). Conclusion: Clinical and laboratory data of COVID-19 patients may help understanding the pathogenesis of the disease and subsequently improve of the outcome of patients by determination of the associated risk factors and recognition of high risk group who are more liable for complications and in hospital death. The present study put an eye on some parameters (laboratory and clinical) that should be alarming signs that the patient is at high risk bad prognosis.

Identification and Genetic Characterization of Blastocystis Species in Patients from Makkah, Saudi Arabia.

Background: Blastocystis species (sp.) are gastrointestinal protozoan parasites with high prevalence rates worldwide. Blastocystis sp. show extensive genetic diversity with 17 different subtypes (STs) described to date. A few studies have investigated the prevalence and STs of Blastocystis sp. in Makkah, Saudi Arabia. Therefore, we aimed in this study to identify and characterize subtypes of Blastocystis sp. in the City of Makkah, Saudi Arabia. Methods: Stool samples were collected from 140 patients who presented to King Abdulaziz Hospital, Hera General Hospital and Modern Medical Center in Saudi Arabia. Different microscopic examination methods of patients' stools and molecular analyses (using primers targeting SSU rRNA gene) were performed to identify and characterize STs of Blastocystis sp. Results: Our microscopic examination of stool samples showed that 96/140 patients (68.6%) had Blastocystis sp. infection. Clinical examination of infected patients revealed that 81 patients were symptomatic, whereas 15 were asymptomatic. Next, we isolated DNA from Blastocystis sp.-positive stool samples followed by PCR amplification of small-subunit ribosomal RNA (SSU rRNA) gene and sequence analysis. Our sequence analysis showed that subtype 3 (ST3) was the most prevalent (53.13%) followed by subtype 1 (ST1) (45.83%), whereas subtype 2 (ST2) was the least prevalent (1.04%). Moreover, our results showed that all three STs resulted in more symptomatic than asymptomatic cases. Finally, we identified novel haplotypes which comprised of 8 ST3, 6 ST1, and one ST2 haplotypes. Conclusion: Our identification of several haplotypes in patients' stools confirms the genetic diversity of Blastocystis sp. and may explain the reported low host specificity and differential pathogenicity of Blastocystis sp. We believe that additional molecular epidemiological and genomic studies are needed to understand the prevalence and pathogenicity of different subtypes in humans and animal hosts.

Genotyping of Taenia hydatigena isolated from sheep and goats in KSA based on Cox1 gene.

Sheep and goats are among other herbivorous animals that serve as intermediate hosts (containing the larval stage Cysticercus tenuicollis) of Taenia hydatigena tapeworm. This infection can lead to serious complications or cause death. The genetic diversity and epidemiological significance of cysticercosis due to T. hydatigena is poorly understood. We examined 11,651 goats and 23,542 sheep slaughtered at the municipal abattoir in Makkah, for C. tenuicollis infection. The resulted DNA sequences were compared with previously available sequences from different hosts. Phylogenetic analysis and Pairwise nucleotide variations of cox1 gene were performed. Sheep and goats revealed infection rates of (4.95%) and (4.75%) respectively. DNA sequence analysis of all isolates from both sheep and goats showed that the total haplotypes number was 7. T. hydatigena population with high haplotypes diversity values. The nucleotide diversity was low, while Tajima's D and Fu's tests were negative (with no statistical significance). The present work will give valuable information regarding the prevalence and implementation of control and prevention measures of C. tenuicollis.

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.

A report on tick burden and molecular detection of tick-borne pathogens in cattle blood samples collected from four regions in Saudi Arabia.

Babesiosis, theileriosis and anaplasmosis are among the most commonly reported tick-borne diseases in cattle and are associated with significant economic losses. Through the present study the researchers aimed to report the presence of various pathogens that cause babesiosis, theileriosis and anaplasmosis in cattle collected from different provinces in Saudi Arabia and to report their phylogenetic relationship. A total of 362 blood samples of cattle along with ticks that were present on the cattle were collected from four regions (Riyadh, Al-Kharj, Al-Hasa and Al-Qassim) of Saudi Arabia. Blood samples were screened by polymerase chain reaction (PCR) for the presence of various Babesia, Theileria and Anaplasma species by amplification of their 18S rRNA and/or 23S rRNA genes. A total of 541 ticks were collected and identified from the cattle. These included Hyalomma anatolicum, Hyalomma dromedarii, Hyalomma impeltatum, Hyalomma excavatum, Rhipicephalus annulatus and Rhipicephalus turanicus. Regarding tick-borne pathogens, the overall prevalence was 1.9 % (7/362) for Theileria annulata, (2/362) 0.6 % for Theileria and (21/362) 5.8 % for Anaplasma ovis. Four of the cattle were found to be co-infected with more than one pathogen (1.1 %). We did not detect any Babesia species in the blood of the studied cattle. Prevalence of the Theileria and Anaplasma species was highest in cattle that resided in Riyadh, followed by cattle from Al-Hasa and Al-Qassim. Representative amplified partial-gene sequences of T. annulata (GenBank accession numbers MK826137-39) and A. ovis (GenBank acc. no. MK 880224) were submitted to GenBank. The presence of ticks on cattle was found to be associated with a high prevalence of Theileria spp. (P = 0.02) and Anaplasma ovis (P < 0.001). We report novel genotypes of T. annulata and A. ovis from cattle in Saudi Arabia and we recommend that molecular surveys are undertaken throughout the country to address the prevalence and geographical distribution of tick-borne infections for their effective diagnosis and treatment.

Cell therapy as a new approach on hepatic fibrosis of murine model of Schistosoma mansoni-infection.

BACKGROUND: Schistosomiasis is an acute and chronic disease of the genus Schistosoma triggered by blood flukes. Schistosomiasis is a disease occurring in, or endemic to, tropical and subtropical regions. A new concept was implemented to deal with schistosomiasis from natural plant sources. Curcumin's common name is Turmeric. Curcumin has proven to be main active component in Curcuma longa L. and has a wide range of anti-phrastic effects. Previous studies have shown the role of bone marrow mesenchymal stem cells (BMSCs) therapy in hepatic fibrosis recovery. OBJECTIVE: The current study was, therefore, intended to examine therapeutic role of BMSCs and Turmeric in murine schistosomiasis mansoni. ANIMALS: Mice were divided into five groups: a negative control group (non-infected non-treated), a positive control group (infected non-treated), a BMSCs treated group; Turmeric treated group, and untreated group. BMSCs derived from male mice were injected intraperitoneally into female mice receiving S. mansoni cercariae through the subcutaneous route. Liver histopathology and immuno-histochemical examinations were evaluated. RESULTS: BMSCs intraperitoneal injection resulted in a significant reduction of liver collagen, granuloma size, and significant increase of OV-6 expression in the Schistosomiasis-treated mice group. There was overall improvement in pathological changes of the liver. Unfortunately, group IV showed a mild improvement in the granuloma size and fibrosis compared to corresponding BMSCs treatment group, although with vacuolated liver cells. CONCLUSION AND CLINICAL RELEVANCE: BMSCs have a regenerative potential in liver tissue histopathology by decreasing liver fibrosis and granulomas. Turmeric, by contrast, could not be used as an anti-fibrotic, according to the findings.

Prevalence and histopathological study on cystic hydatidosis in heart and spleen of goat slaughtered at Makkah, Saudi Arabia

Hydatidosis or echinococcosis is considered to be one of the most common zoonotic diseases of the animals. Infection occurs when intermediate hosts such as camel, cattle, sheep, and goats ingested food or water contaminated with eggs from the definitive host (dog). This is a cross-sectional study which was carried out in one of the biggest abattoirs in Makkah in the west of Saudi Arabia. A total number of 38302 goats were examined and recorded at Makkah abattoirs. The examination had been performed to all slaughtered animals on two organs (spleen and heart) for detection of any hydatid cysts during the period from July 2018 until December 2018. The study included also histopathological tissue evaluation. The total infections number of hydatidosis in goats is 0.23%. The infected hearts were 40.35% whereas the infected spleen was 48.48% subsequently in local animals. The imported animals were 2124, the infected animals in heart were 59.64%, whereas the infected animal involving spleen were 51.51%. Meanwhile, results of histopathological examination had shown that most of the hydatid cysts in goats caused progressive focal pressure and degenerative changes in the surrounding tissue.