Sex determination in juvenile Caretta caretta is difficult because of the lack of heteromorphic sex chromosome and the absence of dimorphic secondary traits. Up till now the most reliable method of gender determination is laparoscopy, that is an invasive and time consuming surgical technique. Testosterone or vitellogenin concentration are proposed as alternative methods but are still not satisfactory. We tried to predict sex of juvenile loggerhead during winter combining the evaluation of body size with the concentration of sexual steroid hormones and vitellogenin. Our results didn’t evidence any correlation between the variables analyzed confirming the general idea of laparoscopy as essential technique in juvenile Caretta caretta sex determination during winter.

The presence of the μ-opioid receptor (MOR) was investigated in the mare oviduct during oestrus and anoestrus, by means of immunoblotting and immunohistochemistry. Immunoblotting analysis showed that the MOR protein is expressed as 65, 50 and 30 kDa forms in the infundibulum and ampulla both in oestrus and anoestrus, while the 30 kDa form is absent in the isthmus. Moreover, different levels of expression were observed along the ampulla in the two periods examined. Immunohistochemistry revealed MOR in the mucosal epithelium, stromal cells, myocytes and blood vessels. Ciliated cells expressed MOR in the apical cytoplasm and, except for the isthmus of oestrous mares, also in the nucleus. Non-ciliated cells showed MOR only in the isthmus segment during oestrus. Stromal cells showed different immunoreactivity along the oviduct segments and during the oestrous and anoestrous phases. The myosalpinx displayed immunostained myocytes in the intrinsic musculature of the ampulla and in the extrinsic and intrinsic musculature of the isthmus without significant differences between anoestrus and oestrus. Blood vessels expressed MOR in endothelial cells and smooth muscle cells in the isthmus myosalpinx of oestrous mares only. In conclusion, these findings show diverse MOR expression in the three segments constituting the oviduct, as well as changes in MOR expression linked to the mare's physiological condition.

Whether a mammal reproduces seasonally or continuously depends mostly on the environment and its effects on the endocrine pattern. Although the dog was the first species to have been domesticated, little information is available on the changes in reproduction that have occurred since. In this study, we evaluated whether environmental stimuli can act as modulators of male gonadal activity in the dog at the latitude of Bari (Italy). Therefore, for 1 year, serum and seminal testosterone (T) concentrations, together with seminal parameters, were recorded monthly and evaluated in relation to environmental variables such as temperature, humidity, and photoperiod. We found that, in temperate regions, the annual serum T profile is not affected by environmental conditions, whereas seminal T profile peaks in October and reaches its nadir in April. The percentage of progressive motile spermatozoa is also dependent on environmental cues. The results support the intuitive idea that recorded data require a proper analysis to be meaningful. In fact, we found that, in the dog, environmental changes appear to affect male gonadal physiology, and this is clear when recorded data are analyzed monthly; in contrast, pooling data into seasonal groups hides monthly environmental variations.

Malassezia spp. may act as opportunistic skin pathogens in humans and animals. Malassezia pachydermatis proliferation and phospholipase production may play a pathogenic role in the occurrence of skin lesions in dogs. This study investigates the presence of mu-opioid receptor (MOR) in M. pachydermatis strains isolated from healthy dogs and dogs with skin lesions and its effects on phospholipase activity (p.a.). P.a. of 64 M. pachydermatis isolates was evaluated using different concentrations of naloxone (Nx), a MOR antagonist. Isolates were divided into Group A (i.e., 40 isolates from 26 dogs with dermatitis) and Group B (i.e., 24 isolates from 12 healthy dogs). The MOR expression was analyzed by Western blot and immunofluorescence. A statistically higher p.a. than that of the controls was found with isolates in Group A at a Nx concentration of 10(-6) M (P<0.05). No isolate in Group B displayed p.a. in either control samples or in the presence of any Nx concentration. Immunoblotting revealed two positive MOR immunoreactive bands of approximately 65 and 98 kDa. MOR expression and localization was also demonstrated by immunofluorescence in isolates from Groups A and B. This study provides the first evidence of MOR expression on M. pachydermatis cell membranes pointing to its possible role in modulating p.a. production in isolates from dogs with skin lesions

The aim of this study was to demonstrate that POMC and the mu opioid receptor (MOR) were expressed in swine reproductive tract and swine spermatozoa. We found that the sow female reproductive tract expresses both POMC and MOR mRNA and MOR protein but, interestingly, the semen shows the highest expression level for both genes and also the highest MOR protein expression. The presence of POMC and MOR (opioid precursor/opioid receptor) transcripts suggests a local synthesis of opioids that could have paracrine and/or autocrine influences during some crucial points of reproduction such as fertilization and pregnancy. Key words: opioid precursor, opioid receptor, reproduction, swine

Leptin has been proven to be crucial in the ovulatory process. Aims of this study are to assess the expression of leptin receptor (Ob-R) in the ovaries of queens at estrus and to evaluate the capability of leptin in modulating ovarian contractility in vitro. Right ovaries underwent immunoblot analysis. Left ovaries were mounted in an organ bath under physiological condition and exposed to murine leptin (10(-6)M). Immunoblot analysis showed that the queen ovary expresses leptin receptor at estrus. Leptin at the dose of 10(-6)M significantly reduced the contractile activity of the ovary. The presence of ovarian Ob-R and leptin inhibitory effects on ovarian contractility suggest leptin implication in the modulation of ovarian activity, as well as in ovulatory disorders.

In aquaculture, unfavourable conditions experienced during early development may have strong downstream effects on the adult phenotype and fitness. Sensitivity to stress, leading to disease, reduced growth and mortality, is higher in larvae than in adult fish. In this study, conducted on sea bream (Sparus aurata), we evidenced the presence of the mu opioid receptor in gametes and larvae at different developmental stages. Moreover, we evaluated the possibility of reducing the effects of artificially produced stress, altering temperature, salinity and pH, by naloxone (an opioid antagonist) and calcium. Results evidenced that mu opioid receptor is present in larvae and in gametes of both sexes and that, during larval growth, its expression level changes accordingly; furthermore, naloxone/calcium association is efficacious in increasing the survival period of treated larvae compared to controls. We conclude that in sea bream rearing, the use of naloxone/calcium against stress can improve fish farming techniques by reducing larval mortality and consequently increasing productivity.

In this work the slaughter-linked plasma modifications of some stress-related hormones in horses subject to standardized butchering procedures were investigated in order to highlight the compromised animal welfare during pre-slaughter handling. During pre-slaughter, animals show strong hardship behavioural patterns, probably due to being under life-threatening conditions. Blood samples from 12 male horses, ageing from 3 to 5 years, were collected before slaughtering in lairage, and during exsanguination after stunning. Catecholamines, cortisol and beta-endorphin concentrations were assessed in plasma samples by EIA. Results show that plasma beta-endorphin concentration did not increase significantly after stunning, while cortisol (P < 0.05) and catecholamines (P < 0.001) increased significantly. The ratio between the plasma level of norepinephrine and epinephrine decreased significantly (P < 0.001) during the time considered for observation underlining a greater involvement of adrenal medulla in the stress response. Moreover these results suggest that, under stress, the release of beta-endorphin could be different from that of ACTH. (C) 2009 Elsevier Ltd. All rights reserved.

Leptin is a Ob gene product secreted mainly by adipose tissue. Several reports showed leptin production by other tissue including the ovary. The action of leptin is mediated upon binding to its receptor widely expressed in reproductive tissues in different species. In fact, there are growing evidences that leptin plays an important role in the modulation of reproductive functions. Therefore, the aim of this study was to evaluate in the queen, the expression of leptin receptor during the functional ovarian cycle and pregnancy. We found that the ovaries of the queen express leptin receptor in all the examined phases. The highest leptin receptor expression was found in the luteal phase (pseudopregnancy, pregnancy) compared to other phases of the cycle (anestrus, proestrus, estrus). The variations in the expression of leptin receptor suggest a likely implication of leptin in the modulation of ovarian activity, in the examined species.

L’esperienza svolta nella ideazione, gestione e realizzazione del progetto pilota “Tutela e Riproduzione del Riccio di Mare (Paracentrotus lividus) specie innovativa per l’acquacoltura” dalla Algesiro srl di Manfredonia e della Facoltà di Medicina Veterinaria Dipartimento di Benessere Animale dell’Università di Bari è un modello per sviluppare la filiera ittica e la collaborazione tra imprese e mondo della ricerca. Classificato al primo posto della graduatoria nella valutazione di merito, avendo colto il senso del bando ed i suoi obiettivi, il progetto è stato condotto con puntualità ed aggiornamento costante rispetto alle scoperte via via effettuate ed alle tecnologie applicate. L’esperienza si colloca nelle buone pratiche che la Regione Puglia intende adottare e replicare perché il settore delle risorse ittiche possa crescere in maniera sostenibile e nel lungo periodo. L’attenzione posta alla tutela dell’ambiente e delle risorse partendo dalla richiesta di mercato del riccio di mare dimostra che è possibile progettare la gestione della filiera ittica bilanciando gli interventi tenendo conto della pluralità degli interessi e della qualità. Il protocollo sviluppato, primo al mondo, è replicabile in ogni angolo del pianeta e la funzione degli echinodermi, sia in allevamento sia in banchi naturali, contribuisce a tutelare l’ecosistema. Con l’auspicio che esperienze positive come questa possano replicarsi attivando rapporti virtuosi tra imprese, mondo della ricerca, istituzioni e ambiente per la crescita del territorio pugliese