Bisphenol A effects on the host Biomphalaria alexandrina and its parasite Schistosoma mansoni.

Bisphenol A (BPA) is one of the most potent endocrine-disrupting chemicals (EDCs) that adversely affect aquatic organisms. The present investigation explored the effects of exposure to BPA at 0.1 and 1 mgL-1 concentrations on the fecundity of Biomphalaria alexandrina, snail's infection with Schistosoma mansoni, and histology of the ovotestis and topographical structure of S. mansoni cercariae emerged from exposed snails. The 24 h LC50 and LC90 values of BPA against B. alexandrina were 8.31 and 10.88 mgL-1 BPA, respectively. The exposure of snails to 0.1 or 1 mgL-1 BPA did not affect the snail's survival. However, these concentrations caused an increase in the reproductive rate (Ro) of infected snails. A slight decrease in egg production was observed in snails exposed to 0.1 mgL-1 BPA after being infected (infected then exposed). However, a significant increase in egg production was noted in snails exposed to 1 mgL-1 BPA after infection with S. mansoni. Histopathological investigations indicated a clear alteration in the ovotestis tissue structure of exposed and infected-exposed groups compared to the control snails. Chronic exposure to BPA caused pathological alterations in the gametogenic cells. SEM preparations of S. mansoni cercariae emerged from infected-exposed snails showed obvious body malformations. From a public health perspective, BPA pollution may negatively impact schistosomiasis transmission, as indicated by the disturbance in cercarial production and morphology. However, it has adverse effects on the reproduction and architecture of reproductive organs of exposed snails, indicating that B. alexandrina snails are sensitive to sublethal BPA exposure.

Molluscicidal and Larvicidal Potency of N-Heterocylic Analogs against Biomophalaria alexandrina Snails and Schistosoma mansoni Larval Stages.

This work describes the synthesis of quinoline-based N--heterocyclic arenes and their biological evaluation as molluscicides against adult Biomophalaria alexandrina snails as well as larvicides against Schistosoma mansoni larvae (miracidia and cercariae). Molecular docking studies were demonstrated to investigate their affinity for cysteine protease protein as an interesting target for antiparasitics. Compound AEAN showed the best docking results followed by APAN in comparison to the co-crystallized ligand D1R reflected by their binding affinities and RMSD values. The egg production, hatchability of B. alexandrina snails and ultrastructural topography of S. mansoni cercariae using SEM were assessed. Biological evaluations (hatchability and egg-laying capacity) revealed that the quinoline hydrochloride salt CAAQ was the most effective compound against adult B. alexandrina snails, whereas the indolo-quinoline derivative APAN had the most efficiency against miracidia, and the acridinyl derivative AEAA was the most effective against cercariae and caused 100% mortality. CAAQ and AEAA were found to modulate the biological responses of B. alexandrina snails with/without S. mansoni infection and larval stages that will affect S. mansoni infection. AEAA caused deleterious morphological effects on cercariae. CAAQ caused inhibition in the number of eggs/snail/week and reduced reproductive rate to 43.8% in all the experimental groups. CAAQ and AEAA can be recommended as an effective molluscicide of plant origin for the control program of schistosomiasis.

Immune responses, DNA damage and ultrastructural alterations of gills in the marine mussel Lithophaga lithophaga exposed to CuO nanoparticles.

Nanoparticle (NP) pollution is a worldwide problem. Copper oxide nanoparticles (CuO NPs) are one of the most used NPs in a variety of applications, which results in their increased release into the marine environment. In the present work, the marine mussel Lithophaga lithophaga was used as a model organism to evaluate the toxic effects of CuO NPs following 28 days of exposure to sub-lethal concentrations (5 and 20 µg/L). The time points were 1 day of exposure to assess the cell viability, phagocytosis in mussel haemocytes and genotoxicity (DNA damage in gills), 1, 14 and 28 days of exposure to evaluate copper concentrations in water and gills, as well as metallothionein concentration in gills, while gill histology and SEM examination were done after 28 days of exposure. The results indicated that the accumulation of CuO NPs in gills increased with concentration and time. Mussel exposure to CuO NPs increased neutral red uptake. However, the phagocytic abilities decreased in haemocytes with increased concentration. CuO NPs caused DNA damage in the gills even at low concentrations (5 µg/L). CuO NPs caused histopathological alterations in gills, such as brown cell accumulation, necrosis, dwarfism of filaments and ciliary erosion. In conclusion, exposure of the mussel L. lithophaga to CuO NPs led to concentration- and time-dependent responses for all the examined biomarkers. Thus, L. lithophaga may be used as a bioindicator organism in the assessment of CuO NP toxicity.

Serological Survey of Babesia bigemina and Babesia bovis in Cattle and Water Buffaloes from Menoufia Province, Egypt.

PURPOSE: Bovine babesiosis causes morbidity in tropical and subtropical countries worldwide. The present study aimed to determine the seroprevalence of Babesia bigemina and B. bovis in cattle and water buffaloes in Menoufia province, where the second-highest population of bovines in Lower Egypt are raised. MATERIALS AND METHODS: A total of 506 blood samples were collected from cattle (N = 262) and water buffaloes (N = 244) in Menoufia province, Egypt. Seroprevalences of B. bigemina and B. bovis in the samples were determined using recombinant Babesia antigen-specific enzyme-linked immunosorbent assays (ELISA). RESULTS: In cattle, the seroprevalences of B. bigemina and B. bovis were 41.60 and 38.17% (37.40 and 35.88% for IgM and 9.54 and 6.11% for IgG), respectively, whereas those of water buffaloes were 35.66 and 31.97% (27.87 and 21.72% for IgM and 15.16 and 15.16% for IgG), respectively. Statistically significant changes in the seroprevalences of the two infective agents were recorded on the basis of region and season of sample collection. CONCLUSION: In conclusion, babesiosis is frequent and presents a threat of an epidemic among bovines in Menoufia province. In turn, control of bovine babesiosis is required because of its potential to detrimentally affect milk and meat production in Menoufia province.

First detection of Anaplasma ovis in sheep and Anaplasma platys-like variants from cattle in Menoufia governorate, Egypt.

Tick-borne diseases are of global economic importance, especially due to the costs associated with disease treatment and productivity losses in livestock. In this study, 244 livestock animals (cattle N = 92, buffaloes N = 86 and sheep N = 66) from Menoufia, Egypt were tested for Anaplasma, Ehrlichia and Babesia species using PCR. Results revealed detection of A. ovis (9.1%) in sheep while Anaplasma spp. (14.1%), A. marginale (15.2%), B. bigemina (6.5%) and B. bovis (5.4%) in cattle. On the other hand, Anaplasma spp. (1.2%), A. marginale (1.2%) and B. bovis (1.2%), were detected in buffaloes. Significantly higher detection rates were observed in cattle for Anaplasma spp. (P = .020), A. marginale (P = .001) and B. bigemina (P = .022) than in buffaloes. Sequence analysis of Anaplasma spp. isolates from cattle, revealed A. platys-like strains. Phylogenetic analyses of the A. platys-like isolates revealed variation among the strains infecting cattle. The A. marginale buffalo isolate, on the other hand, showed some level of divergence from the cattle isolates. This study reports the first detection of A. ovis in sheep and A. platys-like strains in cattle in Menoufia and Egypt at large. The results of the current study provide valuable information on the epidemiology and genetic characteristics of tick-borne pathogens infecting livestock in Egypt.

Molecular and morphological signatures for extreme environmental adaptability of the invasive mussel Brachidontes pharaonis (Fischer, 1870).

Brachidontes pharaonis (Bivalvia:Mytilidae) is one of the most successful Lessepsian migrants. Its extensive populations' expansion and phenotypic plasticity might reshape the Mediterranean biodiversity. Individuals of B. pharaonis were collected from various sites in the Mediterranean Sea and the Red Sea in Egypt. Species-specific primers for Cytochrome Oxidase Subunit 1 gene were designed. They were applied for analysis of mussel's population genetics and assessment of its aquatic environmental DNA (eDNA) abundance. Morphological, allometric and morphometric characteristics were also described. The newly designed primers could efficiently detect the species presence, abundance, and genetic diversity. The Northern Red Sea and north-westward populations exhibited higher nucleotide diversities than southwards. Phylogeny and principal coordinates' analysis (PCoA) detected three geographical categories for B. pharaonis: one of the Indian Ocean, other of the Middle Red Sea and southwards, and the other extends from the Northern Red Sea to the westernmost part of the Mediterranean. Intraspecific differences in the shell shape, colour, and biometrics were noted. The shells were significantly smaller and lighter in rocky habitats than in sandy ones. The morphometric indices and allometry were significantly different between rocky and sandy environments. In general, B. pharaonis genetic and morphological features appeared to contribute much to the species success in versatile habitats.

Morphological and molecular clues for recording the first appearance of Artemia franciscana () in Egypt.

Artemia franciscana is a native species to the New World, and became an exotic species to most parts of the world. The Egyptian hypersaline, continental Qaroun Lake (Fayoum Governorate, Middle of Egypt) is subjected to a gradually increasing salinity rates that approximate or exceed these of seawater. Artemia populations there are known to be parthenogenetic. Yet, these populations started to exhibit abnormal morphologies. Therefore, Qaroun Lake samples of Artemia were subjected to several morphological, biometric, and molecular phylogenetic analyses for accurate species identification and phylogeographic origin approximation. These analyses revealed the existence of the alien sexual species of brine shrimp A. franciscana in Qaroun Lake. The characteristics of the subspherical frontal knob with several spines on the top, ovisac lateral triangular lobe on both sides and its projection together with the biometrics confirmed this species morphotype. DNA barcoding and other molecular analyses based on PCR-based amplification and sequencing of the barcode region of the cytochrome oxidase subunit I gene (COI) exhibited that all the collected samples belong to five haplotypes. Egyptian A. franciscana COI sequences phylogeny and pairwise distances analysis exhibited closer proximity to Latin American strains than to the Northern American ones. A. franciscana presence may be ascribed to the migratory birds present in Qaroun Lake protectorate, since no marine aquaculture activity in Qaroun Lake is known. Therefore, and for the best of our knowledge, this is the first record of the invasive A. franciscana in Egypt.

Molecular and serological prevalence of Babesia bigemina and Babesia bovis in cattle and water buffalos under small-scale dairy farming in Beheira and Faiyum Provinces, Egypt.

In order to determine the molecular and serological prevalence of Babesia bigemina and Babesia bovis, a total of 247 blood samples were collected from cattle and water buffalos in Beheira and Faiyum Provinces in Egypt and examined by nested polymerase chain reaction (nPCR) and enzyme-linked immunosorbent assay (ELISA). In cattle, the prevalence of B. bigemina and B. bovis was 5.30% and 3.97% by nPCR and 10.60% and 9.27% by ELISA, respectively, whereas those of water buffalos were 10.42% and 4.17% by nPCR and 15.63% and 11.46% by ELISA, respectively. Statistically significant differences in the prevalence of the two infections were observed on the basis of age and health status. Sequencing analysis revealed two genotypes for B. bovis spherical body protein-4. In conclusion, the current data provide valuable information regarding the epidemiology of B. bigemina and B. bovis infections in cattle and water buffalos from Egypt, which can be employed in developing future strategies for disease management and control.

Effect of pollution history on immunological responses and organ histology in the marine mussel Mytilus edulis exposed to cadmium.

The effect of previous toxicant exposure (i.e., exposure history) on an organism's response to re-exposure to the toxicant is of considerable interest. The marine mussel Mytilus edulis was collected from reference and polluted sites in southwest England, and groups of mussels from each site were exposed to 20 µg/L CdCl2 for 0, 1, 4, and 8 days and compared with unexposed controls. End points evaluated were tissue metal and electrolyte concentrations, haemolymph chemistry, haemocyte characteristics [counts, neutral red uptake (NRU), and phagocytosis], histology, and expression of metallothionein gene (mt10) expression in digestive glands. Field-collected animals differed by collection site for some end points at time zero, at which time tissue Fe and Pb concentrations were greater and NRU and condition index lower in mussels from the polluted site. Subsequent exposure to cadmium (Cd) in the laboratory caused Cd accumulation mainly in digestive gland, but there were no site-specific effects on tissue trace-metal concentrations. NRU, phagocytosis, and haemolymph Na(+) and K(+) concentrations differed among sites and Cd treatment, but there were no clear trends. Exposure to Cd resulted in lower Ca(2+) concentrations in gill, digestive gland, and haemolymph in animals from the polluted site compared with controls (Kruskal-Wallis, p ≤ 0.05). Lesions, including necrosis, inflammation, and neoplasia, were observed in animals from the polluted site, but the frequency of these lesions appeared to decrease unexpectedly after Cd exposure. Expression of mt10 increased 3-fold in Cd-exposed animals from the polluted site compared with all other groups (Kruskal-Wallis, p = 0.01). We conclude that Cd exposure affected some immune responses in M. edulis, but pre-exposure history influenced toxicological outcomes of Cd exposure in the laboratory.

Tissue injury and cellular immune responses to mercuric chloride exposure in the common mussel Mytilus edulis: modulation by lipopolysaccharide.

Little is known about the immunotoxic effects of sublethal mercury (Hg) exposure in bivalves, and whether or not stimulation of the immune system exacerbates Hg toxicity. The mussel, Mytilus edulis, was exposed to a total inorganic Hg concentration of 50 microgl(-1) as HgCl2 for up to 11 days compared to no added Hg controls. Then the immune functions of haemocytes, haematology, biochemistry and organ histology were assessed. Selected experiments were then repeated in mussels injected with lipopolysaccharide (LPS) to stimulate the immune system prior to Hg exposure. Hg exposure alone caused Hg accumulation mainly in gills and digestive gland, and to a lesser extent in the adductor muscle and haemolymph. This general pattern of Hg accumulation also occurred during the Hg plus LPS experiments. Hg exposure alone caused a transient increase of phagocytosis, but no Hg-dependent effects on neutral red retention or the cytotoxicity of haemocytes. Changes in haemolymph Na+, K+ and glucose levels were negligible during Hg exposure alone compared to controls. However, histological examination showed tissues injuries consistent with inflammation in the gills, digestive gland and adductor muscle during Hg exposure alone. LPS injection had a significant effect (Kruskal Wallis, P=0.02) on circulating haemocytes numbers at the end of the experiment compared to saline injected controls and the Hg+LPS treatment (mean+/-S.E.M., n=6, cells ml(-1)x10(6)); control, 2.1+/-0.08; LPS treated, 3.18+/-0.36, and LPS+Hg, 2.1+/-0.2. The Hg+LPS treatment also caused a transient decline in superoxide dismutase activity in haemocytes (85% decline, Kruskal Wallis, P=0.006) compared to controls. Target tissue inflammation and injury was greatly increased by the effect of Hg exposure with an LPS injection compared to either treatment alone. We conclude that Hg can cause inflammation in mussels in vivo, and that stimulation with LPS can greatly increase Hg-dependent immunotoxicity.