Morphomolecular characterization of Strongylus vulgaris isolated from donkeys with special references to histopathological study on the affected organs.

Equine gastrointestinal tract is infected with Strongylus vulgaris (S. vulgaris) which is highly pathogenic parasite for its harmful effect on cranial mesenteric artery during its migration. So, this study was applied for identification of S. vulgaris in donkeys ultramorphologically and molecularly. In addition to, detection of the pathological effect of larval stage of S. vulgaris on the mesenteric arterial system using histopathology and immunohistochemistry. During the period from September to December; 2019, 60 male and 20 female donkeys at the Giza Zoo was postmortem examined. S. vulgaris adults and larvae were collected from the large intestine and cranial mesenteric arteries (CMAs), respectively. Ultramorphological examination of the collected adults was done using scanning electron microscope (SEM). DNA was extracted from 5 larvae and 6 adults for further conventional PCR studies and sequencing of the internal transcriped spacer 2 (ITS2) gene. The ITS2 gene were amplified and showed bands at 148 base pair (bp). The ITS2 gene nucleotide sequences of all isolates were aligned using Basic Local Alignment Search Tool (BLAST). Histological sections of S. vulgaris affected mesenteric arteries exhibited the presence of the parasite larvae either in the lumen with thrombus formation or attached to the intima. Most of the detected inflammatory cell populations were CD68-positive cells. From these results, it can be concluded that the ribosomal spacers genes could be used as markers for Strongylus species identification in eggs collected from equine feces as a beneficial method of diagnosis. Also, it could be important in disease surveillance, improving preventive measures and developing an effective control strategy.

Evaluation of the cytokines response in buffaloes focused on its milk as a newly emerging indicator tracing for toxocariasis.

This study aimed to evaluate the immune responses and oxidative stress provoked by Toxocara vitulorum infection in buffaloes with special reference to milk parameters as an emerging tool. The use of the milk tool was reported for the first time in tracing T. vitulorum infection in Egyptian buffaloes. Intestine, milk, serum, and liver samples were gathered from flocks in Cairo and Giza districts to evaluate buffalo immune responses provoked by T. vitulorum. The compositional items and somatic cells of milk were monitored. The intestine and milk were evaluated for interleukin IL-1ß, IL-6, and tumor necrosis factor-alpha (TNF-α) by quantitative real-time polymerase chain reaction protocol and the analysis of malondialdehyde (MDA) as an oxidative stress marker. The mean percentages for the total solids, fats, proteins, lactose, salts, pH, and somatic cell count/ml in positive samples were 11.23 ± 0.37, 5.1 ± 0.17, 4.44 ± 0.14, 3.9 ± 0.14, 0.81 ± 0.02, 6.8 ± 0.22, and 4.23 × 106± 1.41 × 105 cells/ml, respectively. A significant difference (p < 0.05) was observed in the mean values of compositional items except for the total protein %, salts %, and pH. For T. vitulorum-contaminated samples, the milk IL-1ß, IL-6, TNF-α, and MDA (nmol/ml) were 7 ± 0.23, 18 ± 0.6, 17 ± 0.56, and 3.7 ± 0.12, respectively (which were less than the values for intestinal cytokines). There is a statistical difference (p < 0.05) between positive and negative samples in the intestinal, milk cytokines, and MDA. This study is an initial investigation of the utilization of intestine and milk cytokines in the evaluation of buffalo toxocariasis.

Evaluation of immune responses and oxidative stress in donkeys: Immunological studies provoked by Parascaris equorum infection.

This study aimed to assess the effects of Parascaris equorum (P. equorum) in infected donkeys through evaluation the oxidative stress and different gene parameters in infected tissues. Fifty donkeys were examined in Giza Zoo abattoir from the period of January to March 2021. Blood and sera samples were collected from each examined donkey. P. equorum were subjected for identification through scanning electron microscope study and the infected tissues were subjected into gene expression analysis using two genes; interleukin 1ß (IL1- ß); and pro-inflammatory cytokines (TNF-α) with assessment of the antioxidant and free radicals released from the animals during the infection. Eighteen donkeys were positive for P. equorum adult or larvae by postmortem examination of the intestine and abdomen with prevalence rate of 36 %. The examined infected donkeys with P. equorum showed significantly higher of Total antioxidant capacity (TAC) levels and the serum malondialdehyde (MDA) 2.45 ± 0.53 than that in non-infected control donkeys. The levels of AST enzyme were 278.54 ± 0.45 while ALT enzyme was 14.97 ± 0.87 which was significantly higher than that of control negative donkeys. The infected donkeys with P. equorum showed significantly upregulation of the TNF-α and IL-1ß which classify according to number of collected worms. The P. equorum infected donkeys exerted at least 100 eggs of parasite in feces. The fecal egg count was marked decreased after treatment with moxidectin. Moxidectin is considered a novel active ingredient that has a marvelous result with high persistency and protection for long time, in addition to, broad spectrum activity and low or no resistance. We recommend the periodical deworming with different molecules as more economic and lifesaving over a single treatment every 12 months parallel with parasitic testing.

Incidence of gastrointestinal parasites in pigeons with an assessment of the nematocidal activity of chitosan nanoparticles against Ascaridia columbae.

In this investigation, the incidence and intensity of gastrointestinal parasites of domestic pigeons were evaluated, additionally, in vitro and in vivo evaluation of the antiparasitic activity of chitosan nanoparticles against the most predominant gut parasite. Therefore, 240 domestic pigeons (160 adults and 80 squabs) obtained from different localities in Giza governorate, Egypt, from February to July 2021, were subjected to parasitological and postmortem examination. The results revealed that 97% of pigeons were vulnerable to single or mixed gastrointestinal parasites. The detected helminths were identified as Capillaria columbae (C. columbae) with a total incidence of (12.5%), Ascaridia columbae (A. columbae) (83.3.%), Heterakis gallinarum (H. gallinarum) (18.7%), Raillietina cesticillus (R. cesticillus) (7.5%), Raillietina echinobothrida (R. echinobothrida) (29%), Choanotaenia infundibulum (C. infundibulum) (22.9%), Davainea proglottina (D. proglottina) (26.6%), and Cotugnia proglottina (C. proglottina) (14.5%). At the same time, the identified protozoan parasites were Trichomonas gallinae (T. gallinae), and Eimeria columbae (E. columbae), with a total incidence of 25 and 79%, respectively. Helminths and Eimeria infections were higher in adults than squabs, while T. gallinae infection was reported with a higher incidence in squabs (62.5%) than adults (6.2%). From our findings, A. columbae was the most predominant gut parasite in the examined pigeons. Thus, it was subjected to in vitro and in vivo treatment with chitosan nanoparticles. Serum and tissue samples were collected from the birds which have been used in the in vitro study to evaluate the oxidative stress markers as malondialdehyde (MDA), Nitric oxide levels and Tumor Necrosis Factor-alpha (TNF-α) and Interleukin-1ß activity also, chitosan nanoparticles- treated worm ultrastructure were determined using scanning electron microscopy (SEM). Finally, we recommend periodic monitoring of pigeon's farm to detect the parasitic infestation, and from our results, we recommend chitosan nanoparticles as a potent nematocidal agent.