Influence of oil-related environmental pollutants on female reproduction.

The petroleum low-weight aromatic hydrocarbons benzene, toluene, ethylbenzene, m/p-xylene, and o-xylene, also known as BTEX, are among the most common hazardous sources of environmental contamination. This paper reviews the available data concerning the effects of BTEX on different aspects of female reproduction, including the fecundity, ovaries, central nervous system (CNS), oocytes, embryos, oviducts, cytogenetics of somatic and generative cells, intracellular signaling systems, and hypothalamic, pituitary and peripheral reproductive hormones. Analysis of the available literature demonstrates that BTEX can exert negative effects on various female reproductive sites, including the CNS-pituitary-ovarian axis, their signaling molecules and receptors, ovarian follicles, corpora lutea, oocytes, embryos, oviducts, ovarian cycles, fertility, and the viability of offspring. These effects could be due to the ability of BTEX to destroy chromosomes, to affect cell metabolism, including the accumulation of free radicals, and to affect the release of hormonal regulators of reproductive processes and intracellular protein kinases.

Evaluation of cytotoxic, oxidative stress, proinflammatory and genotoxic effect of silver nanoparticles in human lung epithelial cells.

Silver nanoparticles are increasingly used in various products, due to their antibacterial properties. Despite its wide spread use, only little information on possible adverse health effects exists. Therefore, the aim of this study was to assess the toxic potential of silver nanoparticles (<100 nm) in human lung epithelial (A549) cells and the underlying mechanism of its cellular toxicity. Silver nanoparticles induced dose and time-dependent cytotoxicity in A549 cells demonstrated by MTT and LDH assays. Silver nanoparticles were also found to induce oxidative stress in dose and time-dependent manner indicated by depletion of GSH and induction of ROS, LPO, SOD, and catalase. Further, the activities of caspases and the level of proinflammatory cytokines, namely interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) were significantly higher in treated cells. DNA damage, as measured by single cell gel electrophoresis, was also dose and time-dependent signicants in A549 cells. This study investigating the effects of silver nanoparticles in human lung epithelial cells has provided valuable insights into the mechanism of potential toxicity induced by silver nanoparticles and warrants more careful assessment of silver nanoparticles before their industrial applications.

Sensitivity of freshwater pulmonate snail Lymnaea luteola L., to silver nanoparticles.

Toxicity of nanoparticles depends on many factors including size, shape, chemical composition, surface area and surface charge. Silver nanoparticles (AgNPs) are likely to enter the aquatic ecosystems because of their multiple applications and pose a health concern for humans and aquatic species. Therefore, we used a freshwater snail Lymnaea luteola L (L. luteola) to investigate the acute toxicity and genotoxicity of AgNPs in a static-renewal system for 96 h. AgNPs caused molluscicidal activity in L. luteola, with 96-h median lethal concentrations (LC50) (48.10 µg L(-1)). We have observed that AgNPs (36 µg L(-1)) elicited a significant (p<0.01) reduction in glutathione, glutathione-s-transferase and glutathione peroxidase with a concomitant increase in malondialdehyde level and catalase in digestive gland of L. luteola. However, a significant (p<0.01) induction in DNA damage was observed by the alkaline single cell gel electrophoresis in digestive gland cells treated with AgNPs for 24 and 96 h. These results demonstrate that silver nanoparticles are lethal to freshwater snail L. luteola. The oxidative stress biomarkers and comet assay can successfully be used as sensitive tools of aquatic pollution biomonitoring.

Potential ultrastructural changes in rat epididymal cell types induced by Boswellia papyrifera and Boswellia carterii incense.

Boswellia papyrifera and Boswellia carterii, known as Arabian incense, diffuses smoke, contaminating the air, which adversely affects human health. Therefore, this study was designed to ascertain the effect of these plants on histopathological and ultrastructure changes in cauda epididymis of Albino rats. Animals were exposed to 4 g/kg body weight of B. papyrifera and B. carterii daily for 120 days along with suitable controls. Our study indicates a significant reduction in epithelial heights. Cells showed signs of degeneration. The ultrastructural study revealed that the cauda epididymis was affected, including its cell types. Furthermore, a decrease in the size of mitochondria, Golgi complex, and both ERs was observed. In all treated groups, plasma fructose decreased considerably, indicating the sign of reduced energy, vital for motility and other sperm functions. The results of this study suggest that these plants systematically affect cauda epididymal cell types and its lumen through its potential toxicity.

The involvement of SIRT1 and transcription factor NF-κB (p50/p65) in regulation of porcine ovarian cell function.

The role of either mTOR system/enzyme sirtuin1 (SIRT1) or transcription factor NF-κB in the direct control of ovarian function has not been estabished. The aim of our in vitro experiments was to examine the involvement of SIRT1 and the p65 and p50 subunits of NFκB in control of porcine ovarian granulosa cell functions and the interrelationships between SIRT1, NFκB (p65, p50) 30 and FSH in the ovary. Monolayers of primary granulosa cells were transfected with gene constructs encoding either SIRT1 or p65 and p50, and thereafter cultured with, or without, addition of FSH. The accumulation of markers of proliferation (cyclin B1 and cyclin-dependent protein kinase Cdc2/p34) and proteins p50, p65 and SIRT1 in the cells was detected by using SDS-PAGE/Western immunoblotting and immunocytochemistry. The secretion of progesterone (P4) and insulin-like growth factor I (IGF-I) was measured by using radioimmunoassay. It was observed that transfection of cells with a SIRT1 gene construct promoted accumulation of proliferation markers, Cdc2/p34, cyclin B1, decreased accumulation of p50 and p65 and stimulated release of P4 and IGF-I. Co-transfection of cells with cDNA p50 and cDNA p65 enhanced the accumulation of SIRT1 and the release of P4 but did not influence the release of IGF-I. Adding FSH to the culture medium stimulated accumulation of both subunits of NF-κB, as well as accumulation of Cdc2/p34, cyclin B1 and release of both P4 and IGF-I. The ability of FSH to promote NF-κB accumulation, the similarity of the main effects of FSH, SIRT1 and NF-κB, as well as the inability of NF-κB to substantially modify the the majority of FSH effects suggest that SIRT1/NF-κB system could be a mediator of FSH action on ovarian cell functions. On the other hand, SIRT1 was able to inhibit NF-κB and to change stimulatory the effect of FSH on NF-κB from stimulatory to inhibitory. This could suggest the existence of negative feedback control of FSH/NF-κB system by high amounts of SIRT1. Our observations (1) confirm the previous data on proliferation, P4 and IGF-I release in ovarian cells and their up-regulation by FSH, (2) demonstrate the presence of SIRT1, NF-κB/p50 and NF-κB/p65 in these cells, (3) show for the first time the involvement of SIRT1 and NF-κB in direct control of proliferation and secretory activity of ovarian cells, (4) represent the first data on interrelationships between FSH, SIRT1 and NF-κB within the ovary.

Reproductive strategies, karyology, parasites, and taxonomic status of Dugesia populations from Yemen (Platyhelminthes: Tricladida: Dugesiidae).

We present new data on the distribution, reproductive strategies, karyology, and taxonomic status of populations of freshwater planarians from Yemen. Nine populations were sampled and significant differences in their reproductive strategies and karyology are reported. The present study presents the first fully documented record of a naturally sexual, diploid (2n = 18) population of a Dugesia species in the eastern part of the Afrotropical region. Morphological characters combined with karyological data suggest that these Dugesia populations from Yemen represent a new species, which is herein described as Dugesia arabica Harrath and Sluys, sp. nov. This new species is mainly distinguishable from other Dugesia species that are distributed exclusively in the Mediterranean basin and in the eastern part of the Afrotropical region by the presence of the following features: well-developed and cone-shaped penis papilla, housing an ejaculatory duct that runs ventrally and has a subterminal and ventral opening; a considerably expanded and folded section of the bursal canal at the level of the oviducal openings; absence of a layer of longitudinal muscles on the copulatory bursa and the bursal canal. Specimens from two populations from Yemen were infested with a gregarine Protozoon.

Kudoa azevedoi n. sp. (Myxozoa, Multivalvulida) from the oocytes of the Atlantic horse mackerel Trachurus trachurus (Perciformes, Carangidae) in Tunisian coasts.

A new species Kudoa azevedoi sp. n. (Myxozoa, Multivalvulida) is described in Trachurus trachurus Linnaeus, 1758 (Carangidae) from fishing harbors in Tunisian coasts using spore morphology and SSU rDNA sequence data. The parasite occurs only in ovaries within oocytes of mature and immature specimens. Spores are quadrate in shape in apical view with rounded edges, having four shell valves and four symmetrical polar capsules. They are of small sizes and measure 3.5±0.41 (3-4.2)×4.5±0.44 (4-5.2) length by width. The polar capsules are pyriform in shape measuring 1.5±0.22 (1.5-2)×0.75±0.14 (0.5-1) µm. Infected oocytes are hypertrophied, whitish colored, and filled with mature spores. Plasmodia are tubular and ramified from the inner membrane toward the center of the oocyte. Phylogenetic analysis based on small subunit ribosomal DNA sequences shows the highest similarity (96%) with the ovary parasite Kudoa ovivora. Some morphological details and spore dimensions support the creation of a new species in the genus Kudoa. Mean prevalence among examined females is of about 55.5%. It varies between localities and length of fish. The present myxosporea is the second Kudoa species reported in fish ovaries.

Erpobdellid leeches (Annelida, Clitellata, Hirudinida) from Tunisia: new records with the description of a new Trocheta species.

Up to now in Tunisia, freshwater Hirudinida are represented by two mainly haematophagous families: Hirudinidae and Glossiphonidae, and a predatory one: the family Erpobdellidae. The present study provides new information on the diversity and taxonomy of erpobdellid leeches. Identification was based, in addition to morphological data, on the length of sperm ducts and the lengths of ovisacs in relation to the neurosomite (ns) and on the shape and size of the male atrium. Five taxons are found. Two subspecies are reported for the first time in the country: Dina punctata punctata Johansson, 1927 and Dina punctata maroccana Nesemann and Neubert, 1994. Tunisian populations of two species, Erpobdella testacea (Savigny, 1820) and Trocheta africana Nesemann and Neubert, 1994, are described, with records of new localities. The new Trocheta tunisiana n. sp. is discovered and described in detail. Trocheta species live in springs in elevated areas while Erpobdella seem to prefer low altitude reservoirs. A comprehensive comparison of the three genera is presented. The disparity between the actual systematics and phylogeny is discussed. This study gives also a detailed distribution of the five species in the north of Tunisia with notes on ecological preference of the genus Dina. Finally a key for the determination of freshwater erpobdellid species from Tunisia is proposed.

Ultrastructure of spermatogenesis and mature spermatozoon of the freshwater planarian Schmidtea mediterranea (Platyhelminthes, Paludicola).

The aim of this study was to characterize for the first time spermatogenesis and spermiogenesis in the freshwater planarian Schmidtea mediterranea using both light and electron microscopy. Starting from the border towards the testis lumen we found types I and II spermatogonia, clusters of primary and secondary spermatocytes, spermatids and free spermatozoa. Light microscope observations show that type I spermatogonia have a large and pale nucleus whereas type II spermatogonia are significantly smaller than the one of type I, and show a darker and central bulky nucleus. At the ultrastructure level, both type I and type II spermatogonia are characterized by a wide nucleus with scanty cytoplasm containing free ribosomes, mitochondria and a dense chromatoid body whereas endoplasmic reticulum and Golgi complex were not observed. The cytoplasm of primary and secondary spermatocytes displays numerous free ribosomes and many endoplasmic reticulum cisternae and Golgi complexes, suggesting that the development of these organelles during spermatogenesis might contribute to the synthesis of hormones and proteins such as testosterone, transcription factors and tubulin. Mature spermatozoa structure closely resembles those of other freshwater triclads with a nucleus, a single fused mitochondrion, a row of cortical microtubules and a pair of flagella conforming to the 9+'1' microtubule pattern described for other Platyhelminthes.

Schmidtea mediterranea phylogeography: an old species surviving on a few Mediterranean islands?

BACKGROUND: Schmidtea mediterranea (Platyhelminthes, Tricladida, Continenticola) is found in scattered localities on a few islands and in coastal areas of the western Mediterranean. Although S. mediterranea is the object of many regeneration studies, little is known about its evolutionary history. Its present distribution has been proposed to stem from the fragmentation and migration of the Corsica-Sardinia microplate during the formation of the western Mediterranean basin, which implies an ancient origin for the species. To test this hypothesis, we obtained a large number of samples from across its distribution area. Using known and new molecular markers and, for the first time in planarians, a molecular clock, we analysed the genetic variability and demographic parameters within the species and between its sexual and asexual populations to estimate when they diverged. RESULTS: A total of 2 kb from three markers (COI, CYB and a nuclear intron N13) was amplified from ~200 specimens. Molecular data clustered the studied populations into three groups that correspond to the west, central and southeastern geographical locations of the current distribution of S. mediterranea. Mitochondrial genes show low haplotype and nucleotide diversity within populations but demonstrate higher values when all individuals are considered. The nuclear marker shows higher values of genetic diversity than the mitochondrial genes at the population level, but asexual populations present lower variability than the sexual ones. Neutrality tests are significant for some populations. Phylogenetic and dating analyses show the three groups to be monophyletic, with the west group being the basal group. The time when the diversification of the species occurred is between ~20 and ~4 mya, although the asexual nature of the western populations could have affected the dating analyses. CONCLUSIONS: S. mediterranea is an old species that is sparsely distributed in a harsh habitat, which is probably the consequence of the migration of the Corsica-Sardinia block. This species probably adapted to temperate climates in the middle of a changing Mediterranean climate that eventually became dry and hot. These data also suggest that in the mainland localities of Europe and Africa, sexual individuals of S. mediterranea are being replaced by asexual individuals that are either conspecific or are from other species that are better adapted to the Mediterranean climate.

[An ultrastructural study of oogenesis in the planarian Schmidtea mediterranea (Platyhelminthe, Paludicola)]. / Étude ultrastructurale de l'ovogenèse chez la planaire Schmidtea mediterranea (Plathelminthe, Paludicole).

The ovary of the freshwater planarian Schmidtea mediterranea has been studied for the first time using both light and electron microscopy methods. The ultrastructure of the ovary revealed two types of cells: accessory cells and germinal cells at various stages of differentiation, distributed along a maturation axis. Initially, oogonia underwent cytoplasm growth due to the development of organelles, such as endoplasmic reticulum, Golgi complex, and mitochondria, which are all involved in the production of cytoplasmic inclusions or yolk globules. It is shown that the chromatoid body and fibrogranular aggregates may participate in the synthesis of vitelline inclusions. When completely mature, the oocytes have become larger, due to the accumulation of nutritive inclusions, which are round in shape and have a paracrystalline structure. These inclusions are interpreted as being yolk globules and may represent a kind of nutritive material for the developing embryo. These ultrastructural features of the ovary agree with the available phylogenetic tree, based on morphological and karyological characters that considers Schmidtea group as a genus and not a subgenus. The presence of sperm between the oocytes suggests that fertilization may occur within the ovary, representing an uncommon condition within the Triclads, in which fertilization usually takes places outside of the ovaries.