Spermatozoan ultrastructure and spermatogenesis in aplousobranch ascidians,with some phylogenetic considerations

Spermatozoa and some stages of spermatogenesis were studied for four species of aplousobranch ascidians. Spermatozoa of Clavelina lepadiformis (Müller) (family Clavelinidae) are plesiomorphous in that they have apical acrosomal vesicles and a moderately elongated, cylindrical nucleus. The elongated mitochondrion is twisted ea. 11/2 times around the nucleus. In their ultrastructural morphology, C. lepadiformis sperm conform in some respects to the least-derived ascidian sperm, those of the phlebobranch ascidian Ciona intestinalis L.; however, the sperm of Clavelina lepadiformis have two apomorphies not shared with those of Ciona sp.: (1) the mitochondrion of Clavelina lepadiformis is long and spiralled along the entire nucleus rather than being comparatively compact and not at all helical; (2) the mitochondrial cristae are elongated parallel to the long axis of the nucleus, whereas in Ciona sp. sperm the cristae are unmodified. In Distaplia sp., Aplidium sp. and Synoicum pulmonaria (Ellis and Solander) the spermatozoa are more derived and consist of a proximal cylindrical and a distal corkscrew-like part. The mitochondrion in Distaplia sp. and Aplidium sp. contains electron-dense material and extends in a long thread around the nucleus. In S. pulmonaria the mitochondrion surrounds the anterior part of the nucleus in mature spermatozoa, and an elongated, dense structure displaying fine striation is enclosed in the mitochondrion in late spermatids. The sperm ultrastructural morphology observed in this study is consistent with the majority view that clavelinids are closest to the ancestral ascidian but is also consistent with other conclusions, particularly that the Cionidae are closest to the stem ascidian.     

No cost of male mating experience on female reproductive success in the almond moth, <Emphasis Type="BoldItalic">Cadra cautella</Emphasis> (Lepidoptera; Pyralidae)

Male copulation experience may have a profound impact on female reproductive success if male reproductive investment declines over consecutive copulations and if females are unlikely to re-mate. Male reproductive investment is particularly interesting in lepidopterans because males produce dimorphic sperm: a fertilizing (eupyrene) and a non-fertilising (apyrene) sperm. In two experiments, we explored the lifetime reproductive investment of male almond moths, Cadra cautella (also known as Ephestia cautella) and examined its influence on female reproductive success. In the almond moth, females re-mate infrequently and males transfer sperm in a spermatophore. Attached to the spermatophore is a large chitinous process, the function of which is unknown. One group of males were permitted consecutive copulations with virgin females and the amount of sperm and size of the spermatophore transferred were compared for all females. We found that the number of both eupyrene and apyrene sperm per ejaculate decreased with his increased mating frequency, while the size of the spermatophore process decreased dramatically after the male’s first copulation. In a second experiment, we allowed males to mate with females throughout their lives and then compared female fecundity and fertilisation success. We found no obvious decrease in female fecundity and fertilisation success with increased male copulation experience, despite the likely reduction in male gametic investment. We discuss potential explanations for the development of this enlarged and elaborate first spermatophore of male almond moths given that it confers no clear fitness advantage to females.     

Biliary secretion,retention and excretion of five 14C-labelled polychlorinated biphenyls in the rat

Five ¹⁴C-labelled polychlorinated biphenyls: 2,4′,5-trichlorobiphenyl, 2,2′,4,5′-tetrachlorobiphenyl, 2,2′,4,5,5′-pentachlorobiphenyl, 2,2′,3,4,4′-pentachlorobiphenyl and 2,2′,4,4′,5,5′-hexachlorobiphenyl were administered orally to bile-cannulated rats. The activity secreted in the bile, excreted in the feces and the urine was determined. Residues of radioactivity in certain tissues and the carcass were also measured.The trichlorobiphenyl showed the highest absorption (93.8%±5.4) from the gastrointestinal tract and biliary secretion of radioactivity (87.6%±6.1 of the dose). The hexachlorobiphenyl showed the lowest absorption and biliary secretion, 28.2%±1.4 and 18.6%±1.3, respectively. The urinary excretion was low and the radioactive residues in the eviscerated carcasses increased with the chlorine content of the biphenyls.     

Keeping the blood flowing—plasminogen activator genes and feeding behavior in vampire bats

The blood feeding vampire bats emerged from New World leaf-nosed bats that fed on fruit and insects. Plasminogen activator, a serine protease that regulates blood coagulation, is known to be expressed in the saliva of Desmodus rotundus (common vampire bat) and is thought to be a key enzyme for the emergence of blood feeding in vampire bats. To better understand the evolution of this biological function, we studied the plasminogen activator (PA) genes from all vampire bat species in light of their feeding transition to bird and subsequently mammalian blood. We include the rare species Diphylla ecaudata and Diaemus youngi, where plasminogen activator had not previously been studied and demonstrate that PA gene duplication observed in Desmodus is not essential to the vampire phenotype, but relates to the emergence of predominant mammalian blood feeding in this species. Plasminogen activator has evolved through gene duplication, domain loss, and sequence evolution leading to change in fibrin-specificity and susceptibility to plasminogen activator inhibitor-1. Before undertaking this study, only the four plasminogen activator isoforms from Desmodus were known. The evolution of vampire bat plasminogen activators can now be linked phylogenetically to the transition in feeding behavior among vampire bat species from bird to mammalian blood. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Determination of pesticides of different chemical classes in drinking water of the state of Santa Catarina (Brazil) using solid-phase microextraction coupled to chromatographic determinations

The evaluation of the concentration of pesticides in drinking water presents a real concern. In this study, a simple and rapid method based on solid-phase microextraction (SPME) followed by gas chromatography–mass spectrometry and electron capture detectors was developed aiming at multiclass determination of 23 pesticides regulated by the Brazilian legislation. The extraction was carried out by direct immersion mode (DI-SPME) using DVB/Car/PDMS fiber coating. In order to improve the extraction efficiency, parameters such as temperature, salting-out effect, and extraction time were optimized. The method was evaluated using drinking water samples spiked with the analytes at different concentrations, and it showed good linearity in the range studied. The values obtained for limits of quantification (LOQ) were below the limits established by Brazilian regulations. Accuracy and precision of the method exhibited satisfactory results, providing relative recoveries from 70 to 123.34% at three spiked levels, and the relative standard deviations ranged from 0.53 to 24.8%. The method was applied in 20 drinking water samples from 13 cities in the State of Santa Catarina, Brazil.