Anatomy of the olfactory system and potential role for chemical communication in the sound-producing Lusitanian toadfish, Halobatrachus didactylus.

The current study investigated the structure and function of the olfactory system of the Lusitanian toadfish, Halobatrachus didactylus, using histology and electrophysiology (electro-olfactogram [EOG]), respectively. The olfactory system consists of a digitated anterior peduncle, of unknown function, containing the inhalant nostril. This then leads to a U-shaped olfactory chamber with the olfactory epithelium-identified by Gαolf-immunoreactivity-on the ventral surface. A large lacrimal sac is connected to this tube and is likely involved in generating water movement through the olfactory chamber (this species is largely sedentary). The exhalent nostril lies by the eye and is preceded by a bicuspid valve to ensure one-way flow of water. As do other teleosts, H. didactylus had olfactory sensitivity to amino acids and bile acids. Large-amplitude EOG responses were evoked by fluid from the anterior and posterior testicular accessory glands, and bile and intestinal fluids. Anterior gland and intestinal fluids from reproductive males were significantly more potent than those from non-reproductive males. Male urine and skin mucus proved to be the least potent body fluids tested. These results suggest that chemical communication-as well as acoustic communication-may be important in the reproduction of this species and that this may be mediated by the accessory glands and intestinal fluid.

Steroidogenesis by testis and accessory glands of the Lusitanian toadfish, Halobatrachus didactylus, during reproductive season.

In teleost fish sex steroids are essential for gonadal function and have marked effects in reproductive and agonistic behavior and in the expression of secondary sexual characteristics. The Lusitanian toadfish, Halobatrachus didactylus, has two male morphotypes: type I males are territorial nest-holders and have large accessory glands while type II males are smaller, have a relatively large testis and small accessory glands. In the present study, the steroidogenic activity of the testis and accessory testicular glands of the Lusitanian toadfish were examined in vitro as well as their presence in urine. The testis of type I males produced 11-ketotestosterone (11KT) and 11ß-hydroxy-4-androstene-3,17-dione (11ßA) from tritiated 17-hydroxyprogesterone, while those of type II males produced testosterone (T) and 11ß,17ß-dihydroxy-4-andosten-3-one (11ßT), but not 11KT. Additionally, the testis and accessory glands of both morphs produced mostly 5ß,3α-reduced and 17,20α-hydroxylated metabolites. Type I, but not of type II, males synthesised 5ß-reduced androgens in their accessory glands. The presence of 11ßA exclusively in the urine of type I males during reproductive season suggests an association with maintenance of secondary sexual characteristics and behavior in this morph. The urine of both types of males contained two 5α-androstane and 5ß-pregnane glucuronides. Among the latter steroids, those that are 17,21-dihydroxylated are potentially metabolites from cortisol and were found only in type I males during the spawning season. The diversity of metabolites produced by the testis and accessory glands and the presence of some in urine is suggestive of a potential role in chemical communication and reproductive behavior.

Effect of dietary l-glutamine supplementation on the intestinal physiology and growth during Solea senegalensis larval development.

The maturation of the intestinal digestive and absorptive functions might limit the amount of absorbed nutrients to fulfil the high requirements of the fast-growing marine fish larva. Glutamine (Gln) has been described to improve intestinal epithelium functions, due to its involvement in energy metabolism and protein synthesis. The purpose of this study was to evaluate dietary 0.2% Gln supplementation on aspects of intestinal physiology, protein metabolism and growth-related genes expression in Senegalese sole larvae. Experiment was carried out between 12 and 33 days post hatching (DPH) and fish were divided into two experimental groups, one fed Artemia spp. (CTRL) and the other fed Artemia spp. supplemented with Gln (GLN). GLN diet had two times more Gln than the CTRL diet. Samples were collected at 15, 19, 26 and 33 DPH for biometry, histology, and digestive enzymes activity, and at 33 DPH for gene expression, protein metabolism and AA content determination. Growth was significantly higher for Senegalese sole fed GLN diet, supported by differences on protein metabolism and growth-related gene expression. Slight differences were observed between treatments regarding the intestinal physiology. Overall, GLN diet seems to be directed to enhance protein metabolism leading to higher larval growth.

Boat noise impacts Lusitanian toadfish breeding males and reproductive outcome.

Anthropogenic noise is a growing threat to marine organisms, including fish. Yet very few studies have addressed the impact of anthropogenic noise on fish reproduction, especially in situ. In this study, we investigated the impacts of boat noise exposure in the reproductive success of wild Lusitanian toadfish (Halobatrachus didactylus), a species that relies on advertisement calls for mate attraction, using behavioural, physiological and reproductive endpoints. Two sets of artificial nests were deployed in the Tagus estuary and exposed to either ambient sound or boat noise during their breeding season. Toadfish males spontaneously used these nests to breed. We inspected nests for occupation and the presence of eggs in six spring low tides (in two years) and assessed male vocal activity and stress responses. Boat noise did not affect nest occupation by males but impacted reproductive success by decreasing the likelihood of receiving eggs, decreasing the number of live eggs and increasing the number of dead eggs, compared to control males. Treatment males also showed depressed vocal activity and slightly higher cortisol levels. The assessment of oxidative stress and energy metabolism-related biomarkers revealed no oxidative damage in noise exposed males despite having lower antioxidant responses and pointed towards a decrease in the activity levels of energy metabolism-related biomarkers. These results suggest that males exposed to boat noise depressed their metabolism and their activity (such as parental care and mate attraction) to cope with an acoustic stressor, consistent with a freezing defensive response/behaviour. Together, our study demonstrates that boat noise has severe impacts on reproductive fitness in Lusitanian toadfish. We argue that, at least fishes that cannot easily avoid noise sources due to their dependence on specific spawning sites, may incur in significant direct fitness costs due to chronic noise exposure.

Reproductive success in the Lusitanian toadfish: Influence of calling activity, male quality and experimental design.

Acoustic signals are sexual ornaments with an established role on mate choice in several taxa, but not in fish. Recent studies have suggested that fish vocal activity may signal male quality and influence male's reproductive success but experimental evidence is lacking. Here we made two experiments to test the hypothesis that vocal activity is essential for male breeding success in a highly vocal fish, the Lusitanian toadfish. We first compared the reproduction success between muted and vocal males. In a second experiment we related male reproduction success with acoustic activity and male quality, including biometric, condition and physiological features. As a proxy for reproductive success we tallied both total number and number of sired eggs, which were correlated. Muting experiments showed that successful mating was dependent on vocalizing. In addition, the number of eggs was positively associated with the male's maximum calling rate. In the second experiment male's reproductive success was positively associated with male condition and negatively related with circulating androgen levels and relative gonad mass, but was not associated with vocal activity. Differences in results may be related with nest design which could have influenced mate choice costs and intra-sexual competition. In the muting experiment nests had a small opening that restrained the large nest-holder but allowed smaller fish, such as females, to pass while in the second experiment fish could move freely. These experiments suggest that a combination of factors, including vocal activity, influence reproductive success in this highly vocal species.

Agonistic sounds signal male quality in the Lusitanian toadfish.

Acoustic communication during agonistic behaviour is widespread in fishes. Yet, compared to other taxa, little is known on the information content of fish agonistic calls and their effect on territorial defence. Lusitanian toadfish males (Halobatrachus didactylus) are highly territorial during the breeding season and use sounds (boatwhistles, BW) to defend nests from intruders. BW present most energy in either the fundamental frequency, set by the contraction rate of the sonic muscles attached to the swimbladder, or in the harmonics, which are multiples of the fundamental frequency. Here we investigated if temporal and spectral features of BW produced during territorial defence reflect aspects of male quality that may be important in resolving disputes. We found that higher mean pulse period (i.e. lower fundamental frequency) reflected higher levels of 11-ketotestosterone (11KT), the main teleost androgen which, in turn, was significantly related with male condition (relative body mass and glycogen content). BW dominant harmonic mean and variability decreased with sonic muscle lipid content. We found no association between BW duration and male quality. Taken together, these results suggest that the spectral content of fish agonistic sounds may signal male features that are key in fight outcome.

A PTH/PTHrP receptor antagonist blocks the hypercalcemic response to estradiol-17beta.

Estradiol (E(2)) increases circulating calcium and phosphate levels in fish, thus acting as a hypercalcemic and hyperphosphatemic factor during periods of high calcium requirements, such as during vitellogenesis. Since parathyroid hormone (PTH)-related protein (PTHrP) has been shown to be calciotropic in fish, we hypothesized that the two hormones could be mediating the same process. Sea bream (Sparus auratus) juveniles receiving a single intraperitoneal injection of piscine PTHrP(1-34) showed an elevation in calcium plasma levels within 24 h. In contrast, injections of the PTH/PTHrP receptor antagonist PTHrP(7-34) decreased circulating levels of calcium in the same period. Intraperitoneal implants of estradiol-17beta (E(2); 10 microg/g) evoked significant increases of circulating plasma levels of calcium and phosphorus and a sustained increases of circulating plasma levels of PTHrP. However, a combined treatment of E(2) and PTHrP(7-34) evoked a markedly lower calcium response compared with E(2) alone. We conclude that PTHrP or a related peptide that binds the PTH/PTHrP receptor mediates, at least in part, the hypercalcemic effect of E(2) in calcium and phosphate balance in fish.

Molecular characterization and expression pattern of zona pellucida proteins in gilthead seabream (Sparus aurata).

The developing oocyte is surrounded by an acellular envelope that is composed of 2-4 isoforms of zona pellucida (ZP) proteins. The ZP proteins comprise the ZP1, ZP2, ZP3, and ZPX isoforms. While ZP1 (ZPB) and ZP3 (ZPC) are present in all species, ZP2 (ZPA) is not found in teleost fish and ZPX is not found in mammals. In the present study, we identify and characterize the ZP1, ZP3 and ZPX isoforms of gilthead seabream. Furthermore, by analyzing the conserved domains, which include the external hydrophobic patch and the internal hydrophobic patch, we show that ZP2 and ZPX are closely related isoforms. ZP proteins are synthesized in either the liver or ovary of most teleosts. Only in rainbow trout has it been shown that zp3 has dual transcription sites. In gilthead seabream, all four mRNA isoforms are transcribed in both the liver and ovary, with zp1a, zp1b, and zp3 being highly expressed in the liver, and zpx being primarily expressed in the ovary. However, determination of the ZP proteins in plasma showed high levels of ZP1b, ZP3, and ZPX, with low or non-detectable levels of ZP1a. In similarity to other teleost ZPs, the hepatic transcription of all four ZP isoforms is under estrogenic control. Previously, we have shown that cortisol can potentiate estrogen-induced ZP synthesis in salmonids, and now we show that this is not the case in the gilthead seabream. The present study shows for the first time the endocrine regulation of a teleost ZPX isoform, and demonstrates the dual-organ transcriptional activities of all the ZP proteins in one species.

Morphometric changes and sex steroid levels during the annual reproductive cycle of the Lusitanian toadfish, Halobatrachus didactylus.

The Lusitanian toadfish has group synchronous oocytes, which grow from November until June-July when they are released probably as a single batch. Blood plasma levels of estradiol-17beta (E(2)) and testosterone (T) increase during vitellogenesis and drop rapidly during final maturation and ovulation, when 17,20beta, 21-trihydroxy-4-pregnen-3-one (17,20beta,21-P) levels increase. The male reproductive apparatus is composed of paired testes and multichambered accessory glands, which secrete mucosubstances and are connected to the spermatic duct. Changes in the gonadosomatic index of males paralleled the females but started to drop slightly earlier. The swimbladder and accessory glands also underwent important seasonal changes in weight reaching a maximum at spawning. T, 11-ketotestosterone (11-KT) and 17,20alpha-dihydroxy-4-pregnen-3-one (17,20alpha-P) were generally low except for a sharp peak in June. 17,20beta,21-P also peaked in June and then declined slowly. 17,20beta-dihydroxy-4-pregnen-3-one (17,20alpha-P) was undetectable in males and females. As with other species of the family two types of males were identified: type I males with smaller testes (ca. 7-fold) and larger accessory glands (ca. 3-fold) and swimbladders than type II. Type I males also had significantly higher (ca. 6-fold) 11-KT levels than type II males. This suggests a role for 11-KT in the development of structures important for reproductive behaviour.

Hormonal control of swimbladder sonic muscle dimorphism in the Lusitanian toadfish Halobatrachus didactylus.

The swimbladder and associated sonic muscle of the Lusitanian toadfish Halobatrachus didactylus increase in size throughout life and are, respectively, 25% and 30% larger in type I (nest-holder) males than females, which may generate sexual differences in sound production. Sexual dimorphism in swimbladder is also evident in the morphological features of sonic muscle fibers. During the breeding season, type I males have smaller myofibril contracting zones surrounded by larger sarcoplasm areas compared with females, possibly an adaptation to speed and fatigue resistance for the production of long mating calls. Type II (floater) males show characteristics that are intermediate, but statistically not significantly different, between type I males and females. Six weeks after castration and androgen (testosterone and 11-ketotestosterone) replacement in type I and type II males there were no alterations either in swimbladder mass or fiber morphology. However, 17beta-estradiol induced a significant decrease in swimbladder mass and sarcoplasm area/myofibril area ratio. Six months after castration there was a clear reduction in the seasonal swimbladder hypertrophy in males and induction of sonic fiber morphological characteristics that resemble those occurring in females (low sarcoplasm area/myofibril area ratio). These results suggest that testicular factors are required to initiate sonic muscle hypertrophy and type I sonic fiber phenotype in H. didactylus, but a specific involvement of androgens has not been completely clarified.

Saúde mental na atenção básica / Mental health in basic health care

A escassez de informações no contexto brasileiro sobre ações de Saúde Mental na Atenção Básica tem sido apontada. Este estudo teve como objetivo identificar o perfil diagnóstico e sócio-demográfico de pacientes assistidos em Saúde Mental no Programa de Saúde da Família em Cachoeira, Bahia. Realizou-se um estudo de corte transversal, examinando 100 (cem) prontuários com diagnósticos psiquiátricos atendidos em ambulatório. Encontrou-se que 54por cento dos atendimentos foram do sexo masculino, sendo 67por cento adultos jovens, 49por cento de procedência urbana e 63por cento sem ocupação. Dos diagnósticos encontrados 37por cento foram de esquizofrenia, transtornos esquizotípicos e delirantes e 18por cento de transtornos afetivos. Os achados sugerem a inclusão das ações de Saúde Mental na Atenção Básica, considerando o elevado percentual de transtornos mentais severos e a precária inserção sócio-econômica desses indivíduos no território onde vivem.

The scarcity of information on mental health in basic health care, within the Brazilian context, has been revealed. The objective of this study was to identify the sociodemographic and diagnostic profile of patients treated for mental health in the Family Health Program in Cachoeira, Bahia. A cross-sectional study was carried out assessing 100 case records with psychiatric diagnoses treated as outpatients. The results illustrated that 54% of the patients treated were male, 67% young adults, 49% living in the urban area, and 63% unemployed. From the diagnoses found, 37% related to schizophrenia, schizotypic and delirious disorders, and 18% related to emotional disorders. The findings suggest the need to include mental health procedures in basic health care, considering the high percentage of severe mental disorders and the precarious socioeconomic insertion of these individuals in the place where they live.

Características sociais e de morbidade dos portadores de transtornos mentais internados em hospitais psiquiátricos e gerais da rede pública do Estado da Bahia / Social and morbidity characteristics of all in-patients of the psychiatric and general hospitals into public netservices in the state of Bahia

Introdução: o movimento social de reforma psiquiátrica brasileira vem apontando, há quase 30 anos, a internação em hospital psiquiátrico como de baixa eficácia, violadora de direitos humanos e propondo a diminuição progressiva desses leitos. A implantação, todavia, de serviços substitutivos ao hospital psiquiátrico não pode desconhecer as características dos usuários internados. Objetivo: estudar as características sociais e de morbidade dos pacientes internados em hospitais psiquiátricos e gerais da rede pública do Estado da Bahia.Metodologia: estudo de corte transversal na população dos pacientes psiquiátricos internados na rede pública, estimados em 97% dos portadores de transtornos psiquiátrico no estado. A coleta foi realizada no prontuário dos pacientes, no período de dezembro de 1997 a julho de 1998. O diagnóstico foi estabelecido pela CID 10. Resultados: foram encontrados 2.069 pacientes internados e divididos em dois grupos: o dos internados em hospitais psiquiátricos, HP (1.939[93,7%]) e dos hospitais gerais, HG (130[6,3%]). A rede pública estudada constou de 19 unidades de internação, das quais duas definitivamente fechadas e apresentou uma relação de 1 leito/5.326 habitantes. As unidades públicas foram as que mais leitos tiveram dentre o total encontrado (385[15,7%]). A comparação entre os grupos, HP e HG, apontou particularidade significantes: no HG a população foi mais jovem (31,7 anos), tinha maior percentual no II grau .