Dogs as a source for the spreading of enteric parasites including zoonotic ones in Giza Province, Egypt.

To investigate the impact of domestic and stray dogs on the transmission of zoonotic and other parasites to humans in contact with them, fecal samples were collected from 80 domestic dogs that presented at a clinic with health disturbances and 220 randomly selected stray dogs housed in shelters. The parasitological examination of these samples revealed infection by six zoonotic and four non-zoonotic parasites in varying percentages. The zoonotic parasites included Ancylostoma caninum, Toxocara canis, Dipylidium caninum, Echinococcus granulosus, Cryptosporidium species, and Giardia cysts and trophozoites. The other parasites included Toxascaris leonina, Trichuris vulpis, Taenia species eggs, and Isospora canis oocysts. The infection rate was higher in stray dogs (60%) than in domestic dogs (40%). Infected dogs in both groups were generally unhealthy, with poor body condition recorded in 13.8% of domestic dogs and 63.6% of stray dogs. The infection rate was higher (92%) among shelter workers than among domestic dog owners (66.7%). Giardia assemblages A and D from dogs and assemblage A from humans, as well as two isolates of Cryptosporidium canis (C. canis), one from dogs and the other from humans, were submitted in the GenBank with the accession numbers OQ870443, OQ870444, and OQ919265 for Giardia and OQ917532 & OQ915519 for C. canis of dogs & human, respectively. In conclusion, domestic and stray dogs play an essential role in transmitting zoonotic parasites to humans in contact with them, and regular deworming and strict hygienic measures are recommended to minimize their impact on human health.

Trichinocidal activity of a novel formulation of curcumin-olive oil nanocomposite in vitro.

Curcumin-olive oil nanocomposite (CO-NC), a novel formulation of nano-curcumin, was produced and characterized. By evaluating the death rate and DNA damage inflicted on adult Trichinella spiralis (T. spiralis) worms using the comet test and Scanning electron microscopy (SEM) analysis, the effectiveness of the substance against these worms was assessed in vitro. The mortality effects of CO-NC on the parasite adult worms were increased with the upgrading in the concentration and exposure time from 1 to 24 h using concentrations from 10 to 100 ppm. LC50 was determined to be 10.0 ppm/18 h, 20.0 ppm/9 h, 40.0 ppm/6 h, 80.0 ppm/2 h, and 100.0 ppm/1 h, while LC100 was 40.0 ppm/24 h, 80.0 ppm/12 h, and 100.0 ppm/6 h. The comet assay was utilized to examine DNA damage in control and dead worms exposed to varying doses. A direct correlation (P ≤ 0.05) was found between the increase in CO-NC dose and the degree of DNA damage as indicated by alterations in DNA % in the tail segment, tail length (µm), tail moment (µm), and olive tail moment with the control samples. The sub-epidermal layer was detached, the cuticle was partially sloughed off, and the usual creases, ridges, and annulations were altered in the T. spiralis exposed worms. As a result, the tested new trichinocidal drug formulation of nano-curcumin on an oil base was confirmed to be an efficient, secure, and environmentally friendly alternative. The medication has the potential to severely and irreversibly harm the DNA and ultrastructural morphology of adult worms.

Virulence of Babesia bigemina in infected cattle (Bos taurus): Molecular and immunological studies.

This study examined 400 tick-infested cattle from the following four governorates in Egypt: Faiyum, Beni Suef, Giza, and Minya. These cattle were examined for blood parasites between January 2021 and April 2022. The infected cattle were classified into four groups based on tick infestations and clinical signs. Blood was drawn for assessing oxidative stress markers as well as for parasitological examination and molecular analysis of the 18S rRNA gene of Babesia bigemina (B. bigemina). We performed a comparison of the levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) between B. bigemina-infected blood samples and non-infected blood samples used as negative controls. Babesia spp. infection increases hemolysis, which in turn increases oxidative stress marker levels and cell-mediated immune response.

Detection and molecular characterization of Trichomonas gallinae recovered from domestic pigeons in Egypt.

Trichomonas gallinae is a protozoan parasite that causes canker in pigeons. Squabs (young pigeons) are frequently infected with T. gallinae and can die because of the infection, while adult pigeons can act as carriers showing no clinical signs. In the present study, 50 squabs, up to 1-month-old, were purchased from pigeon markets in different regions of the Giza governorate, Egypt. Direct wet mount preparations of the oral excretions of the squabs (mouth wash) and Giemsa staining revealed that 64% (32/50) were positive for T. gallinae. Experimental infection of ten squabs with 103 T. gallinae trophozoites/ml resulted in oral lesions on the mouth, tongue, and soft palate, with the presence of yellowish-white nodules (cheese-like) in the oral cavity on the sixth day post-infection in all squabs. A subset of five samples were cultured in modified Diamond's media, their DNA was extracted, and a portion of the ribosomal internal transcribed spacer region (ITS1/5.8S/ITS2) was amplified by polymerase chain reaction (PCR) followed by sequencing. Phylogenetic analysis of the five isolates revealed 64-91% homology with some reference isolates circulating in Egypt and related countries.

Microfluidic-Based Formulation of Essential Oils-Loaded Chitosan Coated PLGA Particles Enhances Their Bioavailability and Nematocidal Activity.

In this study, poly (lactic-co-glycolic) acid (PLGA) particles were synthesized and coated with chitosan. Three essential oil (EO) components (eugenol, linalool, and geraniol) were entrapped inside these PLGA particles by using the continuous flow-focusing microfluidic method and a partially water-miscible solvent mixture (dichloromethane: acetone mixture (1:10)). Encapsulation of EO components in PLGA particles was confirmed by Fourier transform infrared spectroscopy, thermogravimetric analysis, and X-ray diffraction, with encapsulation efficiencies 95.14%, 79.68%, and 71.34% and loading capacities 8.88%, 8.38%, and 5.65% in particles entrapped with eugenol, linalool, and geraniol, respectively. The EO components' dissociation from the loaded particles exhibited an initial burst release in the first 8 h followed by a sustained release phase at significantly slower rates from the coated particles, extending beyond 5 days. The EO components encapsulated in chitosan coated particles up to 5 µg/mL were not cytotoxic to bovine gut cell line (FFKD-1-R) and had no adverse effect on cell growth and membrane integrity compared with free EO components or uncoated particles. Chitosan coated PLGA particles loaded with combined EO components (10 µg/mL) significantly inhibited the motility of the larval stage of Haemonchus contortus and Trichostrongylus axei by 76.9%, and completely inhibited the motility of adult worms (p < 0.05). This nematocidal effect was accompanied by considerable cuticular damage in the treated worms, reflecting a synergistic effect of the combined EO components and an additive effect of chitosan. These results show that encapsulation of EO components, with a potent anthelmintic activity, in chitosan coated PLGA particles improve the bioavailability and efficacy of EO components against ovine gastrointestinal nematodes.

Animal hydatid cyst genotypes as a potential substitute for human hydatid cyst as a source of antigen for diagnosis of zoonotichydatidosis.

The Diagnosis of hydatidosis is still an unsolved issue due to difficulties in obtaining of patient's hydatid cyst appropriate for antigen extraction. This study evaluated the suitability of HC protoscolices somatic antigens (HCPsS-Ag) fractions from animal origin to substitute that extracted from HC of patients in diagnosis of hydatidosis using enzyme-linked immunoelectro-transfer blot and Enzyme-linked immunosorbent assay (ELISA). Eight fractions in HC-G6 from patients react specifically versus HC-G6 infected patient's sera. Five of them (28, 32, 38, 59 and 89 Kilo Dalton (KDa) and two of them (28 KDa and 45 KDa) reacted versus HC-G1 and HC-G4 infected sheep and equine sera, respectively. Six fractions in HCPsS-Ag-G1 of sheep react versus HC-G1 sheep infected sera, four (28, 32, 52 and 58 KDa) and two of them reacted versus HC-G6 and HC-G4 infected patient and equine sera, respectively. Two fractions only in HCPsS-Ag-G4 of equine reacted versus infected human and sheep sera. This fraction displayed the same degree of ELISA value versus different infected sera with a significantly perfect classification for kappa agreement and non-statistically significant difference (p ≥ 0.05) for ELISA Optical density values of the positive samples without cross-reaction versus other parasites antibodies in sera. HCPsS-Ag from HC genotypes that developed in humans and animals as HC-G6 and HC-G1 can substitute each other for diagnosis of infection than antigens extracted from non-zoonotic HC-G4. The fraction at 28 KDa is the only fraction that can be extracted from any animals HC and used in diagnosis of zoonotic hydatidosis.

Nematocidal Effects of a Coriander Essential Oil and Five Pure Principles on the Infective Larvae of Major Ovine Gastrointestinal Nematodes In Vitro.

The anthelmintic effects of extracted coriander oil and five pure essential oil constituents (geraniol, geranyl acetate, eugenol, methyl iso-eugenol, and linalool) were tested, using larval motility assay, on the third-stage larvae (L3s) of Haemonchus contortus, Trichostrongylus axei, Teladorsagia circumcincta, Trichostrongylus colubriformis, Trichostrongylus vitrinus and Cooperia oncophora. Coriander oil and linalool, a major component of tested coriander oil, showed a strong inhibitory efficacy against all species, except C. oncophora with a half maximal inhibitory concentration (IC50) that ranged from 0.56 to 1.41% for the coriander oil and 0.51 to 1.76% for linalool. The coriander oil and linalool combinations conferred a synergistic anthelmintic effect (combination index [CI] <1) on larval motility comparable to positive control (20 mg/mL levamisole) within 24 h (p < 0.05), reduced IC50 values to 0.11-0.49% and induced a considerable structural damage to L3s. Results of the combined treatment were validated by quantitative fluorometric microplate-based assays using Sytox green, propidium iodide and C12-resazurin, which successfully discriminated live/dead larvae. Only Sytox green staining achieved IC50 values comparable to that of the larval motility assay. The cytotoxicity of the combined coriander oil and linalool on Madin-Darby Canine Kidney cells was evaluated using sulforhodamine-B (SRB) assay and showed no significant cytotoxic effect at concentrations < 1%. These results indicate that testing essential oils and their main components may help to find new potential anthelmintic compounds, while at the same time reducing the reliance on synthetic anthelmintics.

Highly specific Electrochemical Sensing of Pseudomonas aeruginosa in patients suffering from corneal ulcers: A comparative study.

Pseudomonas aeruginosa is the most common pathogenic gram-negative bacteria causing corneal ulcers globally. In severe cases, often after trauma and eye injury, corneal destruction progresses rapidly and may be completed within 24-48 h causing blindness. In our preliminary work, we have established an ultrasensitive polyaniline (PANI)/gold nanoparticles (Au NPs)/indium tin oxide (ITO) modified sensor for rapid detection of pyocyanin (PYO) in P. aeruginosa infections with a linear range from 238 µM to 1.9 µM and a detection limit of 500 nM. In the present study, we evaluated the efficiency of the established modified electrochemical sensor in the diagnosis of P. aeruginosa in 50 samples collected from patients suffering from corneal ulcers. The obtained results were compared with the results gained by the screen-printed electrode, conventional techniques, automated identification method, and the amplification of the 16 s rRNA gene by PCR as a gold standard test for P. aeruginosa identification. We have found that the electrochemical detection of PYO by square wave voltammetry technique using PANI/Au NPs modified ITO electrode was the only technique showing 100% agreement with the molecular method in sensitivity, specificity, positive and negative predictive values when compared with the SPE, conventional and automated methods.

Rapid and highly sensitive detection of pyocyanin biomarker in different Pseudomonas aeruginosa infections using gold nanoparticles modified sensor.

Successful antibiotic treatment of infections relies on accurate and rapid identification of the infectious agents. Pseudomonas aeruginosa is implicated in a wide range of human infections that mostly become complicated and life threating, especially in immunocompromised and critically ill patients. Conventional microbiological methods take more than three days to obtain accurate results. Pyocyanin is a distinctive electroactive biomarker for Pseudomonas aeruginosa. Here, we have prepared polyaniline/gold nanoparticles decorated ITO electrode and tested it to establish a rapid, diagnostic and highly sensitive pyocyanin sensor in a culture of Pseudomonas aeruginosa clinical isolates with high selectivity for traces of pyocyanin when measured in the existence of different interferences like vitamin C, uric acid, and glucose. The scanning electron microscopy and cyclic voltammetry techniques were used to characterize the morphology and electrical conductivity of the constructed electrode. The determined linear range for pyocyanin detection was from 238 µM to 1.9 µM with a detection limit of 500 nM. Compared to the screen-printed electrode used before, the constructed electrode showed a 4-fold enhanced performance. Furthermore, PANI/Au NPs/ITO modified electrodes have demonstrated the ability to detect pyocyanin directly in Pseudomonas aeruginosa culture without any potential interference with other species.

The prevalence of Gasterophilus intestinalis (Diptera: Oestridae) in donkeys (Equus asinus) in Egypt with special reference to larvicidal effects of neem seed oil extract (Azadirachta indica) on third stage larvae.

Gasterophiline larvae are of veterinary and medical importance caused specific equine intestinal myiasis. Gasterophilus intestinalis (Botfly larvae) had a wide geographical distribution. The present study explores the prevalence rate of G. intestinalis 3rd stage larvae in Egypt from January- December 2017; besides, in vitro trials to control of this larvae and evaluation of this trial using Scanning Electron Microscope (SEM) and histopathology of treated larvae. In the present study, the 3rd larval stage of G. intestinalis was found in clusters in the epithelium of the investigated stomach and infested with prevalence rate 97.2%. The highest collected numbers of larvae were found in two months; March and August (570 & 520 larvae) and lowest numbers (200 larvae) were collected in October, November, and December. The calculated LC50 and LC90 values of neem seed extract were 707.9 ppm and 1090.7 ppm. The different alteration was recorded after exposure to oil extract which showed some destruction on cuticle surface as folded and corrugated cuticle, destruction of maxillae with pits on its surface, disfigure and irregularity of cephalic spines. Histopathology of exposed G. intestinalis larvae showed different changes as thinning of cuticle at the different level (exocuticle, endocuticle, cell layers), degeneration of epithelial cells of the gut and different degree of necrosis was described. Life cycle of G.intestinalis was followed up after treatment with neem seed extract.

The prevalence and intensity of external and internal parasites in working donkeys (Equus asinus) in Egypt.

AIM: This study aims to record and update the prevalence and intensity of external and internal parasites in working donkeys (Equus asinus) in Egypt during the period from January to December 2017. MATERIALS AND METHODS: A total of 120 donkeys (10 donkeys each month) were examined at Giza zoo abattoir through bimonthly visits. The examined donkeys were obtained from five governorates (Giza [20], Fayoum [40], Beni Suef [30], Monofia [20], and Assiut [10]). The animals were grouped according to age and sex. RESULTS: All examined donkeys were positive with at least one internal or even external parasitic species. The overall prevalence rate was 100%. A total of 11 helminths species (10 nematodes and 1 metacestode); 7 protozoal and 7 arthropod species were collected. The number of each parasite and intensity of infection with regard to age and sex was recorded. CONCLUSION: All examined donkeys were infected with parasites with an overall prevalence of 100%. So, we recommended following up and continuous treatment of such diseased animal.