Immunological and molecular evaluation of zoonotic metacercarial infection in freshwater fish: A cross-sectional analysis.

Improperly cooked fish, carrying active metacercariae (MCs), can pose a significant risk for transmitting fish-borne zoonotic trematodes (FBZTs) to human consumers. This study aimed to enhance our understanding of FBZTs by conducting a comprehensive cross-sectional analysis involving various fish species, such as Nile tilapia (Oreochromis niloticus), African catfish (Clarias gariepinus), and red-belly tilapia (Tilapia zillii). These fish specimens were collected from distinct Egyptian governorates, specifically Giza, Kafr al-Shaykh, and Fayoum. The recovered flukes from experimentally infected domestic pigeons were identified as Prohemistomum vivax, Haplorchis pumilio, and Pygidiopsis genata based on morphological features. Furthermore, the identity of the retrieved adult flukes was confirmed using three species-specific primers for PCR amplification and sequencing analysis of the ITS rDNA region and have been deposited in GenBank with the following accession numbers: P. vivax (OR291421.1 and OR291422.1), P. genata (OP099561.1), and H. pumilio (OM439581.1-OP090510.1). Quantitative real-time PCR targeting the immunological genes Tumor Necrosis Factor-alpha (TNF-alpha) and Interleukin-1 (IL-1Β) was employed to compare the cellular immune response between infected with EMCs and uninfected O. niloticus. The results indicated a significant increase in TNF- and IL-1Β levels in FBZTs-infected vs un-infected fishes. Importantly, the presence of adult flukes and EMCs led to substantial histological alterations in both experimentally infected pigeons and naturally infected fish tissues. These changes included the necrosis of fish muscle bundles and a pronounced inflammatory reaction with muscular necrosis in the digestive tracts of experimentally infected pigeons.

A multidisciplinary study on Clinostomum infections in Nile tilapia: micro-morphology, oxidative stress, immunology, and histopathology.

Yellow grub disease, caused by Clinostomum metacercaria, is an endemic zoonotic infection in freshwater fish, responsible for Halzoun syndrome transmitted through the consumption of raw infected fish. This study aimed to conduct a multidisciplinary investigation integrating detailed morphology, oxidative stress, immunology, and histopathology alteration to advance our understanding of Clinostomum infection. In this annual study, 400 Nile tilapia (Oreochromis niloticus) were collected from the Nile River at Al Bahr Al Aazam, Giza Governorate to assess Clinostomum infection prevalence. Of the examined fish, 160 individuals (40.0%) harboured larval Clinostomum infections. Clinostomum metacercariae were observed in various anatomical locations, with 135 fish (33.8%) in buccal cavities, 21 fish (5.25%) in gill chambers, and 4 fish (1.0%) on the skin. Infection intensity ranged from 2 to 12 cysts per fish, averaging 5 cysts, notably with skin infections characterized by a single cyst in each fish. Macroscopic encysted metacercariae were collected from buccal cavities, gills, and skin. Micro-morphology revealed distinct features in C. complanatum, including an elliptical oral sucker with collar-like rings and large sensory papilla-like structures, contrasting with the absence of these features in C. phalacrocoracis. Oxidative stress, assessed through malondialdehyde (MDA) and nitric oxide levels, revealed an elevation in MDA to 35.13 ± 6 nmol/g and nitric oxide to 25.80 ± 3.12 µmol/g in infected fish. In infected fish, MHC-I gene expression increased approximately 13-fold, MHC-II peaked at 19-fold, and IL-1ß significantly upregulated by 17-fold, compared to control levels. Histopathology detailed associated lesions, such as cyst encapsulation and eosinophilic infiltration. Clinstomiasis and its impacts on fish hosts offer crucial insights to control this emerging fish-borne zoonotic disease, threatening wildlife, aquaculture, and human health.

Ultra-structure, genetic characterization and Immunological approach of fish borne zoonotic trematodes (Family: Heterophyidae) of a redbelly tilapia.

Fish-borne zoonotic trematodes (FBZT) are highly significant zoonotic trematodes that can infect humans by eating raw or undercooked fish harboring active metacercaria. In this investigation, FBZT was found in samples of widely cultivated redbelly tilapia (Tilapia zillii) obtained from the Fayum governorate. Encysted metacercaria (EMC) infection was identified in fish belonging to the heterophyid family morphologically. The prevalence of heterophyid EMC was 30.5%. EMC was identified and implemented in a subsequent study on domestic pigeons (Columba livia domestica) carried out to allow adult flukes of Pygidiopsis (P.) genata; P. summa; and Ascocotyle (A.) pindoramensis species in their small intestine. This study presents the first report that combines ultra-structure, molecular approach of three species of heterophyid flukes, ultra-structure using transmission electron microscope in P. genata, and the study of host immunological responses and associated cytokines during Pygidiopsis species infection of pigeons in Egypt. Using Quantitative Real-time PCR (qRT- PCR), the gene expression levels of six cytokines (IL-1, IL-2, IL-6, IL-10, IFN-γ and TGF-ß3) were assessed. The molecular confirmation of P. genata, P. summa, and A. pindoramensis have a registration in the GenBank under accession number MT672308.1, OR083433.1, and OR083431.1, respectively. Throughout the infection, the gut produced cytokines in considerably variable amounts. As a result of the Pygidiopsis species infection in pigeons, our data showed distinctive cytokine alterations, which could aid in figuring out the immunological pathogenesis and host defense mechanism against this infection. This study focused on different types of fish-borne trematodes, particularly the zoonotically important ones.

The phylogenetic position and analysis of Renicola and Apharyngostrigea species isolated from Cattle Egret (Bubulcus ibis).

Cattle Egret (Bubulcus ibis) is one of the most well-known herons in Egypt. It is called the friend of the farmer because it benefits farmers and helps them get rid of insects and worms. It acts as a reservoir for many diseases. Few researchers have discussed the significance of parasitic diseases that affect this wild bird and may lead to mortalities among the population especially the importance of vital organs such as kidneys. Therefore, this study aimed to spotlight parasitic infection-affected herons in Egypt and consider the risks to this beneficial bird. During this study, 23 Bubulcus ibis were captured after their death from Abou Rewash Giza Governorate, Egypt, during the period from February to September (2022). Renicola species (spp.) and Apharyngostrigea spp. are two important digenean parasites that were recovered from the kidneys, and small intestine of the heron Cattle Egret (Bubulcus ibis) with an infection rate of (17.2%) and (11.8%) respectively. Histopathological techniques were used to assess tissue alterations while light microscopy and molecular assays were used to assess the parasites. The parasites' morphological and morphometrical characteristics, as well as polymerase chain reaction and sequencing assays (mitochondrial sections), were investigated for the first time in Egypt. These parasites were given in-depth illustrations and drawings. The distinctive qualities of the two species were discussed. As the first record from Egypt, the nucleotide sequences discovered in this work have been uploaded into the GenBank database (accession numbers: OR021986 and OQ955829). Microscopically, the renal blood vessels had vasculitis, necrosis, and other degenerative alterations. Further research analyzing the health of various heron spp. and environmental deterioration can help to close information gaps about the interactions between parasites, their hosts, and environmental health.

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.

Oxidative stress markers, immune-regulating cytokines, and the pathological evaluation of sheep co-infected with Oestrus ovis and Coenuruscerebralis.

This study aimed to evaluate the immune response using the immune-regulating cytokines as (IL-1ß; MHC-I and MHC-II) associated with co-infected sheep Oestrusovis and Coenurus cerebralis as well as oxidative stress markers (malondialdehyde "MDA" and nitric oxide "NO"). So; sheep samples from different regions in Egypt showed different neurological signs, were examined for detection of the cause of the nervous manifestations. Moreover, the O. ovis and C. cerebralis cysts were collected and identified using scanning electron microscopy. The brain tissues were evaluated for different immunological genes such as MHC-I, MHC-II, and Interleukin-1ß activity using quantitative real-time PCR (qRT-PCR) techniques, where the infected sheep showed higher MHC-I gene expression (10-fold), higher MHC-II gene expression (peaked at 25-fold), and higher IL-1ß gene expression (14-fold) than the control group. The MDA level was significantly increased. Also, stress marker (nitric oxide) levels were significantly higher in infectedsheep than in negative control one. During gross pathology, migrating larvae of O. ovis and C. cerebralis were noticed In such areas, hemorrhages and patches of clotted blood were noticed. cysts with prominent protoscolices were also observed and were attached to the caudal region near the cerebellum.

Prognostic Value of LINC-ROR (rs1942347) Variant in Patients with Colon Cancer Harboring BRAF Mutation: A Propensity Score-Matched Analysis.

Emerging studies show that long intergenic non-protein coding RNA, regulator of reprogramming (LINC-ROR) is aberrantly expressed in several types of cancer, including colon cancer (CC). LINC-ROR intronic variant rs1942347 may impact gene regulation and disease phenotype. We aimed to explore the potential association of LINC-ROR (rs1942347) with the clinicopathological features and outcome of CC cases. Archived FFPE (n = 180) CC samples were enrolled. Taq-Man allelic discrimination PCR was used for genotyping in propensity-matched cohorts with/without positive staining for mutant BRAF protein after eliminating confounders bias. The rs1942347*A allele variant was associated with high pathological grade, larger tumor size, distant metastasis, and mortality. Multiple logistic regression analysis adjusted by sex and BRAF mutation showed A/A genotype carriers to have 3 times more risk of early onset of cancer (OR = 3.13, 95%CI = 1.28-7.69, p = 0.034) than T/T genotype carriers. Overall analysis showed that rs1942347*A allele carriers had higher risk of mortality under heterozygote (OR = 2.13, 95%CI = 1.08-4.35, p = 0.003), homozygote (OR = 5.0, 95%CI = 1.69-14.29, p = 0.003), dominant (OR = 3.33, 95%CI = 1.20-9.09, p = 0.003), and recessive (OR = 2.63, 95%CI = 1.37-5.0, p = 0.011) models compared to T/T allele carriers. Stratified analysis by BRAF status revealed that the ancestor T/T allele conferred protection in BRAF mutant CC patients and was associated with a 73-93% reduced risk of mortality under heterozygote/homozygote comparison models. Using Kaplan-Meier curves, carriers of the A/A genotype had shorter survival times than T/T cohorts. The univariate Cox regression model revealed that the A/A genotype was associated with a 3.5 times greater mortality risk than the T/T genotype. However, after adjustment by multiple Cox regression analysis, the risk was insignificant. In conclusion, this is the first study identifying the potential association of the LINC-ROR (rs1942347) variant with CC prognosis.

Parasitological and histopathological examination of Cocktail lovebirds infected with Eimeria aratinga (Apicomplexa: Eimeriidae).

Coccidiosis is a devastating worldwide disease and is considered a dreadful disease in lovebirds. Indeed, a problem has been appeared cocktail lovebirds kept in a private pet birdhouse in Sheikh Zayed City, Giza, Egypt, in the shape of blood-tinged diarrhea, birds huddled together and showing signs of inappetence, ruffled feathers, unable to fly, general weakness and emaciation associated with high mortalities. Therefore, this study aimed to diagnose and find a suitable treatment to overcome such problems. To achieve this aim, blood and droppings samples were collected from infected and healthy birds for parasitological and hematological examinations, and tissue samples were collected from freshly dead birds for postmortem and histopathological examinations. A treatment trial was adopted on 50 infected birds and 25 healthy and parasitological negative birds and groups were classified as follows: group 1) 25 infected birds treated with Diclazuril, group 2) infected birds treated with Coccicure, and group 3) 25 birds kept as control negative reference birds. The parasitological identification revealed the presence of Eimeria aratinga (E. aratinga) oocysts in the infected bird intestine. Finally, we concluded that E. aratinga is a serious protozoon parasite infesting lovebirds revealing severe clinical signs, high mortalities, histopathological changes in the intestine and alteration in blood parameters. Diclazuril is an effective drug in treating E. aratinga in cocktail lovebirds.