In this study, we performed transcriptome analysis in the cecum tissues of negative control untreated non-challenged (NC), positive control untreated challenged (PC), and Bacillus subtilis (B. subtilis) fed challenged chickens (BS + ET) in order to examine the underlying potential therapeutic mechanisms of Bacillus based probiotic feeding under an experimental Eimeria tenella (E. tenella) infection. Our results for clinical parameters showed that birds in probiotic diet decreased the bloody diarrhea scores, oocyst shedding, and lesion scores compared to positive control birds. RNA-sequencing (RNA-seq) analysis revealed that in total, 2509 up-regulated and 2465 down-regulated differentially expressed genes (DEGs) were detected in the PC group versus NC group comparison. In the comparison of BS + ET group versus PC group, a total of 784 up-regulated and 493 down-regulated DEGs were found. Among them, several DEGs encoding proteins involved in immunity, gut barrier integrity, homeostasis, and metabolism were up-regulated by the treatment of probiotic. Functional analysis of DEGs also revealed that some gene ontology (GO) terms related with immunity, metabolism and cellular development were significantly affected by the exposure of probiotic. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that the DEGs in the cecum of B. subtilis-fed challenged group were mainly participated in the pathways related with immunity and gut barrier integrity, included mitogen-activated protein kinase (MAPK) signaling pathway, toll-like receptor (TLR) signaling pathway, extracellular matrix (ECM)–receptor interaction, tight junction, and so on. Taken together, these results suggest that Bacillus based probiotic modulate the immunity, maintain gut homeostasis as well as barrier system and improve chicken metabolism during E. tenella infection.

In Egypt, tick-borne diseases pose a significant threat to human and animal health, and the threat to dromedaries (Camelus dromedarius), the country's dominant camelid species, is of particular concern. These animals are frequently infested with ticks, and may thus develop tick-borne diseases or become reservoirs of tick-borne pathogens. However, there is a paucity of data on tick infestation in Egyptian camels, especially in the south of the country. Accordingly, we aimed to determine the prevalence of tick infestation in southern Egyptian camel populations (in Luxor and Aswan governorates), and identify the hemoprotozoan parasites carried by camel-infesting ticks. Camels were checked for ticks during veterinary examination at quarantine and household checks, and ticks were collected from infested camels for species identification using morphological examination and PCR analyses. Tick and hemoprotozoan species were identified using Basic Local Alignment Search Tool analysis with subsequent confirmation in phylogenetic analyses. All camel-infesting ticks belonged to the species Hyalomma dromedarii, and were clustered with ticks of this species previously found in Egypt in a phylogenetic tree based on the 16S rRNA gene. Molecular analysis targeting the 18S rRNA gene revealed the presence of hitherto undetected hemoprotozoan parasites, Colpodella spp., in 30/297 (10.1 %) camel-infesting ticks. In phylogenetic analysis, these Colpodella spp. were highly homologous (94-98.6 %) with Colpodella spp. previously deposited in GenBank with accession numbers OQ540590Q, MH208621, and GQ411073, which relate to Colpodella spp. previously detected from Haemaphysalis longicornis, Rhipicephalus haemaphysaloides, and humans in China. PCR analyses with spherical body protein-4 (SBP-4) gene-specific primers revealed Babesia bovis in 16/297 (5 %) of camel-infesting ticks, however, Babesia bigemina and Theileria annulata were not detected. Here, we report the first detection of Colpodella spp. in H. dromedarii in Egypt. Further epidemiological studies are needed to assess the risk to camels and humans, and the transmission dynamics. Based on the high tick infestation rates in Egyptian camels and the identification of previously unreported protozoan hemoparasites in ticks, we consider that the dromedary should be subject to surveillance as a sentinel species for tick-borne diseases in Egypt. Our findings underline the need for surveillance and collecting data on lesser known pathogens circulating in camel-infesting ticks, as part of a public health strategy for dealing with tick-borne diseases in Egypt.

Dengue virus (DENV) threatens global health; specific antiviral drugs are required to combat it. Such anti-DENV therapeutics can be rapidly developed by repositioning the drugs approved for other indications. This study investigated six medications of different classes drawn from a library of molecules. In silico analyses were performed to determine potential binding affinity for the DENV non-structural protein NS3 protease and NS5 RNA-dependent RNA polymerase (RdRp). Of the six candidates, galidesivir and tadalafil showed the highest binding affinities for the DENV NS3 protease and NS5 RdRp, with tadalafil demonstrating the highest binding affinity. Galidesivir and tadalafil substantially suppressed viral replication in DENV replicon cells without inducing cytotoxicity and showed half-maximal inhibitory concentrations of 10 μM and 2.56 μM, respectively. Both galidesivir and tadalafil effectively suppress DENV infection in human hepatoma and baby hamster kidney cells, and tadalafil demonstrates protease-inhibitory activity. In an AG129 mouse model of DENV infection, both galidesivir and tadalafil reduced viral loads in the serum, with tadalafil producing a notable reduction by day four. Both drugs markedly suppressed DENV replication in the hepatic tissue. Histopathologically, both galidesivir- and tadalafil-treated mice showed alleviation of DENV-induced lesions in the spleen and liver, indicating the potential therapeutic effects of these drugs. These findings highlight the potential of repositioning galidesivir and tadalafil as effective anti-DENV therapies with low cytotoxicity, meeting the urgent global need for new therapeutic agents against this pathogen.

Ticks serve as vectors for numerous pathogens affecting human and animal health and harbor various microorganisms, including Francisella-like endosymbionts (FLEs), whose potential pathogenicity remains uncertain. In Egypt, data on FLEs, particularly in Southern Egypt, are limited, despite the importance of tick-borne pathogen surveillance in livestock. This study represents the first report of FLEs in Southern Egypt and the first detection of FLEs in Rhipicephalus annulatus in Egypt. A total of 726 ticks infesting cattle and camels were collected from various locations in Southern Egypt. Ticks were identified using both morphological and molecular techniques based on the 16S rRNA gene, and FLE detection was performed using PCR assays targeting a fragment of the Francisella 16S rRNA gene. Francisella DNA was detected in 60.1 % (288/479) of the examined ticks, with the highest prevalence in Hyalomma marginatum (100 %, 4/4), followed by Hyalomma dromedarii (74.8 %, 243/325), and R. annulatus (27.3 %, 41/150). Sequencing of 19 positive samples revealed that H. dromedarii (n = 13), H. marginatum (n = 2), and R. annulatus (n = 4) carried FLE sequences with 99-100 % similarity to strains previously identified in ticks worldwide. Given the high prevalence of FLEs in livestock-associated ticks, further research should focus on characterizing these strains, assessing their role in tick-borne pathogen ecology, and evaluating potential implications for human and animal health.

Equine theileriosis represents one of the main and serious health problems affecting equines industry globally, that is caused by tick-borne protozoan parasite called T. equi. This study aimed to assess the sensitivity of three diagnostic tools named: microscopic examination of a blood smear, conventional PCR, and Real-Time PCR (qPCR) to detect T. equi among equine population (n = 116) raised in Giza Governorate, Egypt. Microscopic examination of Giemsa-stained blood smears revealed the infection of 16.4% (19/116) of examined equines by T. equi while conventional PCR and qPCR revealed that 29.3% (34/116) and 43.1% (50/116) of examined equines were infected with T. equi respectively. Our results demonstrated that the qPCR had the highest sensitivity (100%) followed by conventional PCR (68%) while microscopic examination had the lowest sensitivity (38%). Furthermore, the negative predictive value (NPV) of qPCR was the highest (100%) compared to conventional PCR and microscopical examination (80.49% and 68.04% respectively) which revealed that all negative cases detected by qPCR were certainly correct compared to the other two diagnostic assays. Therefore, it is highly recommended to incorporate PCR diagnostic assays (conventional PCR and qPCR) alongside microscopic examination to evaluate the epidemiological status of equine theileriosis.