Cytochrome P450, acetylcholinesterase and gonadal histology for evaluating contaminant exposure levels in fishes from a highly eutrophic brackish ecosystem: the Orbetello Lagoon, Italy.

Biochemical markers and ovarian histology were investigated in prespawning females of grass goby (Zosterisessor ophiocephalus) and grey mullet (Mugil cephalus) collected, respectively, in late spring and summer 2000 in four sites of a highly eutrophic brackish ecosystem of central Italy, the Orbetello Lagoon. Exposure to chlorinated and aromatic hydrocarbons was evaluated in fish livers by the somatic liver index (SLI) and by measuring 7-ethoxyresorufin-O-deethylase (EROD) and benzo(a)pyrene monooxygenase (BaPMO) activities. Acetylcholinesterase (AChE) activity was measured in brain and gills to evaluate exposure to organophosphates (OPs) and carbamates (CBs). The gonad somatic index (GSI) was used to confirm ovarian maturation and ovarian histology was investigated as a potential biomarker for environmental effects. Samples from the Western Basin, near a sewage treatment plant (STP) off the town of Orbetello, showed higher SLI values and higher EROD and BaPMO activities than those collected from the Ansedonia Canal (AC) in the Eastern Basin (p<0.05) and respect to those from reference sites: the Albegna River (AR) Delta for grass goby and the Nassa Canal (NC), connected with the sea, for grey mullet both located in the Western Basin as well. Low brain AChE activity was observed in both species from the reference sites (AR and NC) in association with the presence of anomalies in developing oocytes: unexpectedly small in grass goby and irregular disintegrated cytoplasm in grey mullet. The results indicate that the Western Basin is more polluted than the Eastern Basin particularly in the Orbetello where the sewage treatment plant may be a source of aromatic and chlorinated compounds while the Albegna River and the Nassa Canal may be sources of OPs and CBs.

Ascorbic acid induces spectrin reorganization in bull epididymal spermatozoa.

The acrosome, a complex organelle, plays a key regulatory role in the sperm-egg interaction. We have previously shown that ascorbic acid affects both motility and spectrin protein patterns in sperm. In this study, we further characterized the changes in spectrin in sperm challenged with ascorbic acid, using SDS-PAGE, western blots, and immunofluorescence. Ascorbic acid shifts spectrin to a higher-molecular-weight species based on western blot studies. This shift in the spectrin band correlates with a striking series of changes in spectrin immunofluorescence patterns. Upon ascorbic acid challenge, spectrin localization changes, eventually resulting in the formation of vesicles. These vesicles can reach sizes up to five times the original volume of the sperm cell and sometimes show multiple spikes. These findings indicate that a novel process is taking place in the acrosome upon ascorbic acid challenge and suggest that the cytoskeleton may be a useful target for studying and hopefully controlling the sperm-egg interaction.

Submicroscopic mathematical evaluation of spermatozoa in assisted reproduction. 4. The bovine fertilization (Notulae seminologicae 10).

In this paper we apply a modification of the formula of Baccetti et al. (1995) in the evaluation of submicroscopical characteristics of bull spermatozoa used in assisted reproduction. In the present experiment sperm quality is proposed as a useful parameter in predicting the success of fertilization. Our results demonstrate that the percentage of spermatozoa devoid of submicroscopic defects, according to the particular Bayesan formula proposed by us, is clearly correlated with the result of artificial insemination. In fact, the parameter concerning sperm quality obtained in variously successful donors shows a large correlation with fertility power. The synthetic parameters observed are therefore a good tool in the prediction of sperm power in artificial fertilization. The evaluation is mainly concerned with the quality of the acrosomal characters, the status of the chromatin, the shape of mitochondria, the position of the postacrosomal sheath, the perinuclear space and the axonemal pattern. All these characters are expressed with different means in ejaculates. All these data confirm that submicroscopic-mathematical evaluation offers a convincing and reliable diagnosis based upon sperm structure and functions such as acrosomal reaction and cell motility. It has been also demonstrated that sperm quality is a major factor in the success of artificial insemination and it is clearly revealed in the integrity of most of sperm organelles.

Correlation between male fertility and acrosin-like protease activity in rats treated with Spartium junceum.

Intraperitoneal injections of extracts of Spartium junceum in adult male rats were shown to reduce the rate of fertility and acrosin enzyme activity. For the latter the drug effects could be divided into two phases: in the first stage a significant increase in proteolytic activity was observed in all experimental animals examined; and in the second stage, there was a general decrease of up to 50% in enzyme hydrolytic properties. There is a notable difficulty in correlating the acrosin activity and the data obtained from mating tests. The effect of drugs on the activity of another acrosomal protease, benzamidine-resistant protease, was assayed. A significant decrease in this enzyme was noted though the modality was different from that of acrosin. Extrapolating the data of the two co-extracted acrosomal proteases, the ratio of acrosin-like protease [E] and benzamidine-resistant protease [E]x, it was possible to correlate the value expressed to the rate of pregnancy. The implications of this finding is discussed although the exact molecular basis of the phenomenon is uncertain.

Evaluation of the antifertility activity of the broom Spartium junceum in the mammalian male.

Male adult rabbits and rats treated with Spartium junceum showed a significant decrease in fertility, demonstrated by a lower number of pregnancies. The target of the drug seems to be the acrosomal protease system, the activity of which appears greatly reduced, while the morphology of testicular cells and epididymal spermatozoa is only partially affected. The antifertility effect is completely reversible.

HIV particles detected in spermatozoa of patients with AIDS.

In this paper the Authors describe the presence of HIV particles on and in mature spermatozoa either ejaculated by AIDS patients or incubated in vitro with HIV. Both kinds of spermatozoa have particles localized around the sperm organelles. In the first case, the nucleoid of the virus can be enveloped by a membrane-like coat or be devoid of it and form buddings in the plasma membrane. In the in vitro infected spermatozoa, only membrane enveloped nucleoids are present, and no process of budding can be found. The Authors conclude considering that the spermatozoa of the AIDS patients can be penetrated by the virus particles in different moments of their life, and show the HIV particles in different stages of their cycle: some of them have freshly penetrated the sperm, and are still contained in a membrane-like coat, others are replicated and are budding through the sperm plasma membrane. On the contrary, in vitro infected spermatozoa have only freshly penetrated virus particles, and lack buddings and membrane-free nucleoids. The presence of the HIV virus in spermatozoa is substantiated by labelling with monoclonal or polyclonal anti-HIV antibodies.

Morphogenesis of the decapitated and decaudated sperm defect in two brothers.

The "decapitated sperm" defect, found in both of two sterile brothers, may be assumed to have a genetic origin. The present material suggests that the term "decapitated spermatozoa" is not exact, because detached heads and tails were found in the brothers' ejaculate that could be regarded as "decapitated tails" and "decaudated heads." The present report describes frequent, more or less advanced stages of detachment. Both heads and tails showed a normal structure in which only the postnuclear region was deficient, lacking basal plate and implantation fossa. A break at a different level of the midpiece, and therefore three kinds of separation, were observed. The defect, according to the present research, must originate in the testicular region, whereas the detachment occurs in the epididymis.

Localization of acrosomal enzymes in Arthropoda, Echinodermata and Vertebrata.

The presence and localization of acrosin and hyaluronidase have been detected by immunocytochemistry in animals belonging to different phyla and characterized by the different structure of the acrosomal complex. Acrosin is widely distributed from Insecta to Echinodermata and Vertebrata; hyaluronidase has a similar diffusion, but seems to be absent from an Insect and from Echinodermata. The presence of extraacrosomal layer and perforatorium seems not to be related to that of the two enzymes, that are absent from Teleost species devoid of acrosome or having only a pseudoacrosome.

Crater defect in human spermatozoa.

This report describes the "crater defect" in human spermatozoa, a malformation that consists of a nuclear and acrosomal invagination present in 100% of the cells, whereas tail structure and motility are fairly normal. The defect occurs during spermiogenesis. A possible concomitance with abnormalities in the microtubular apparatus involved in the sperm molding is discussed.