Florfenicol induces malformations of embryos and causes altered lipid profile, oxidative damage, neurotoxicity, and histological effects on gonads of adult sea urchin, Paracentrotus lividus.

The frequent occurrence of antibiotics in the aquatic environment has engendered negative impacts on non-target organisms. The effects of the veterinary antibiotic florfenicol (FLO) during the embryo-larval development of the sea urchin, Paracentrotus lividus was assessed using four increasing concentrations (1, 2, 5 and 10 mg/L). Furthermore, FLO toxicity to adults was investigated through the analysis of oxidative damage, histopathological alterations, lipid metabolism and acetylcholinesterase activity following an exposure period of 96 h. FLO induced embryotoxicity with estimated EC50 values of 5.75, 7.56 and 3.29 mg/L after 12 h, 24 h and 48 h, respectively. It generated oxidative stress assessed as lipid peroxidation in gonads despite the increased antioxidant activity of catalase (CAT). Neurotoxicity was also evident since the AChE activity significantly decreased. Moreover, FLO affected the lipid metabolism by increasing saturated fatty acid (SFA) and monounsaturated fatty acid proportions (MUFA), except in the group exposed to 5 mg/L. The increase in polyunsaturated fatty acid (PUFA) levels and docosahexaenoic acid (DHA, C22:6n-3) proportions were noted with all FLO concentrations. Eicosapentaenoic acid (EPA, C20:5n-3) decreased, while arachidonic acid (ARA, C20:4n-6) increased in sea urchins exposed to 5 and 10 mg/L FLO. Histopathological alterations of gonadal tissues represent an additional confirmation about the toxicity of this antibiotic that might decrease the reproductive performance of this species. Nevertheless, even if reproduction of sea urchins would be partially successful, the embryotoxicity would compromise the normal development of the embryos with consequences on the population.

Effects of rare earth element samarium doped zinc oxide nanoparticles on Mytilus galloprovincialis (Lamarck, 1819): Filtration rates and histopathology.

BACKGROUND: Doping was reported to improve the photo catalytic performance, antioxidant, antibacterial and other biological properties of nanoparticles. While, improving the nanoparticle properties, doping could change toxicity profile to living organism. Hence, the aim of this work was to assess the effects of samarium doped zinc oxide nanoparticles (Sm doped ZnO NPs) on the edible mussel Mytilus galloprovincialis. METHODS: Sm doped ZnO nanoparticles were characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR) techniques. 156 mussels were exposed during 7 days to a low, intermediate and high concentration of Sm doped ZnO NPs (0.5, 1 and 1.5 mg/L, respectively). The filtration rates were assessed after 1 and 2 h. Histopathological alterations were determined in gills, digestive glands and gonads using a quantitative analysis. RESULTS: The filtration rates decreased in all individuals exposed to Sm doped ZnO NPs, a significant decrease was noted with the low and intermediate concentration (0.5 and 1 mg/L) of Sm doped ZnO NPs after 1 and 2 h, respectively. The histopathological index (Ih) estimated for gills, digestive glands and gonads showed differences depending on the organ and the nanoparticle concentration. The highest Ih were reported for digestive glands and female gonads exposed to the intermediate concentration (1 mg/L) of Sm doped ZnO NPs. As for gills and male gonads, the highest Ih were noted with the high concentration (1.5 mg/L) of Sm doped ZnO NPs. CONCLUSION: Results from this study revealed the toxicity of Sm doped ZnO NPs in Mytilus galloprovincialis gills, digestive glands and gonads. The toxicity induced by this nanoparticle varies depending on the organ and the concentration.

Placobdella nabeulensis sp. nov. (Hirudinea: Glossiphoniidae), a new glossiphoniiform leech from Palearctic North Africa.

BACKGROUND: Herein, we describe a new species of turtle blood-feeding leech, Placobdella nabeulensis sp. nov. from Palearctic North Africa (Tunisia and Algeria). The new species is described based on detailed morphological analyses using light and scanning electron microscopes. RESULTS: Apart from the detailed morphology of the atrium, morphological features alone do not sufficiently separate the species from congeners due to the absence of distinct diagnostic characters. Therefore, we turned to molecular data to better distinguish this new species from other members of the genus and establish a basis for its genetic separation. Four DNA fragments were successfully amplified, including mitochondrial COI and 12S rDNA, as well as nuclear 28S rDNA and histone H3. We then provided the molecular descriptor of the taxon, based on redundant diagnostic nucleotide combinations in DNA sequence alignment within the Folmer region. Results of the phylogenetic analysis and species delimitation methods (ABGD, ASAP, and bPTP) based on the COI locus support the species rank of the Tunisian-Algerian Placobdella. CONCLUSIONS: The new species is most closely related to the European species Placobdella costata (Fr. Müller, 1846) and the present study indicates that Placobdella nabeulensis sp. nov. has likely been confused with the European counterpart in several previous studies. This article is registered at www.zoobank.org under urn:lsid:zoobank.org:pub:4A4B9C1D-2556-430F-8E4B-0CE99F2012F5.

Ultrastructural study of spermatogenesis and sperm in the African medicinal leech Hirudo troctina Johnson, 1816 (Annelida, Hirudinida).

This paper presents the process of spermatogenesis in the leech Hirudo troctina Johnson, 1816 using light, fluorescent and transmission electron microscopy. At the onset of spermatogenesis in testes, the pear-shaped spermatogonia divide mitotically without full cytokinesis and as a result isogenic groups are formed (clusters, clones) with 2, 4, 8, 16, 32, 64, 128 spermatogonia and, finally, 256 primary spermatocytes occur. The final meiotic divisions of spermatocytes give rise to clones with 1024 spermatids. There are hundreds of developing germ-line clones in each testis. In each clone, the male germ cells divide in full synchrony and they are in the same phase of spermatogenesis. During complex spermiogenesis each spermatid becomes a filiform spermatozoon with a helicoid nucleus, which is characterized by the presence of a long acrosome with two regions - anterior and posterior, which are followed by a helicoid nucleus, a midpiece with only one mitochondrion and a long flagellum. Our results were compared to those on other clitellate annelids that have been studied to date, especially to sperm formation in Hirudo medicinalis Linnaeus, 1785. Only minor differences were found in the length and the diameter of different organelles and the number of spermatids in germ-line clones.

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.

An ultrastructural study of the ovary cord organization and oogenesis in Erpobdella johanssoni (Annelida, Clitellata: Hirudinida).

The aims of the present study were to analyze the ovary cord structure and oogenesis in Erpobdella johanssoni under light, fluorescent and transmission electron microscopy and to compare the obtained results with other clitellate annelids, especially with other arhynchobdellid leeches. Each of the paired ovaries is composed of the ovary wall (ovisac) and several (7-8) short, cone-shaped ovary cords. The ovary cords are of the "Erpobdella" type, i.e. they are short and polarized and five zones containing germ cells at consecutives stages of their development can be distinguished along their long axis. One, huge somatic cell (the apical cell), oogonia and premeiotic germ cells occur at the tip of the apical part of the ovary cord - zone I. Below, in zone II germ cells enter meiosis, whereas in zone III only a few cells continue meiosis and gather nutrients (oocytes), while the rest become nurse cells. In zone IV, huge vitellogenic oocytes form protuberances on the surface of the cord, and degenerating germ cells were observed at the base of the ovary cord (zones IV and V). The germline cells form syncytial cysts in zones I-III. The germline cysts have broadly the same architecture as in the ovaries of all of the clitellate annelids that have been described to date. Each germ cell in a cyst has only one cytoplasmic bridge connecting it to the common cytoplasmic mass - the cytophore. The cytophore is poorly developed, and it has the form of thin, long cytoplasmic strands. The presence of two categories of germ cells suggests a meroistic mode of oogenesis. The germline cysts are closely associated with somatic, follicular cells. There are two subpopulations of follicular cells: one envelops the growing oocytes, while the second is distributed between other germ cells. The entire ovary cord is additionally enveloped by a layer of somatic cells with a spongy appearance - the spongiosa cells. A characteristic feature of vitellogenic oocytes is the condensation of the chromosomes into a karyosome. Fully grown oocytes are excluded from the ovary cords and float freely in the ovisac lumen.

Batracobdella algira Moquin-Tandon, 1846 (Hirudinida: Glossiphoniidae)--morphometric analysis and internal morphology.

There has been very few papers on leeches of Batracobdella algira. The aim of the presently reported study was to provide the measurements and, for the first time, description and characteristics of the reproductive and digestive systems of B. algira. The material used in this study was collected from frogs, Rana saharica Boulenger, 1913, in April 1987 by H. Hotz in Béja in Tunisia (11 individuals) and by R. Ben Ahmed in 2008 from the skin of Bufo mauritanicus Schlegel, 1841 in Nabeul in Tunisia (18 individuals). The relative length of the specimens ranges between 1.48 and 4.07. The digestive system of leeches has a standard structure for the species from the family Glossiphoniidae. Male and female gonopores are separated by two annuli. Ovisacs are as long as 5 neurosomits. The characteristic features of the male reproductive system of Batracobdella are sperm ducts, which are twisted in a small area and placed near to atrium. This study will hopefully contribute to a better understanding of the host-parasite relation.