Morphology, endocrinology, and environmental modulation of gonadal sex differentiation in teleost fishes

Successful reproduction by an adult depends on the normal ontogenesis of the gonads, a complex process of cellular and histological differentiation that starts early in life. This process is theoretically predetermined by genetic factors and includes sensitisation of the bipotential gonads to endogenous endocrine factors prior to, during and even after commitment to maleness or femaleness. However, young fish are relatively vulnerable to a host of environmental (physical and chemical) factors that can affect this endogenous endocrine axis, disturbing or even overriding the putative developmental pathway. This sexually lability can be exploited to our advantage for the production of monosex fish populations of the most valuable sex for food production or aquarium fish trade. On the other hand, it represents also a potential path for undesirable influences from endocrine-disrupting chemicals and climatic factors, particularly environmental temperature. This paper provides a detailed account of the early histological process of gonadal sex differentiation, with special reference to gonochoristic species, and reviews the criteria employed to positively identify ovarian and testicular differentiation. It also reviews the development of endocrine competence and sensitivity of the differentiating gonads to exogenous influences in the context of the relative stability of genotypic sex determination in various fish species. Sex differentiation in some species seems to be under strong genetic control and may not require endogenous sex steroid production. Conversely, reliance on endogenous sex steroids for gonadal differentiation is observed in other species and this phenomenon is apparently associated with a higher incidence of environment (mainly temperature)-labile sex differentiation. This revised version was published online in July 2006 with corrections to the Cover Date.     

Detection of post‐F1 fish hybrids in broodstock using molecular markers: approaches for genetic management in aquaculture

Herein, we have developed molecular markers for nuclear genes to use in multiplex‐PCR and PCR‐RFLP, with the goal of characterising hybrid lines derived from crosses between pintado Pseudoplatystoma corruscans and cachara P. reticulatum. These markers, together with others described previously, were used to perform molecular identification analyses as genetic subsidies for Brazilian aquaculture. These analyses were performed due to the problems of high mortality in the offspring reported by the aquaculturist. From a total of 16 broodstock samples, 13 were genetically identified as hybrids; surprisingly, nine of these hybrids were found to be post‐F1 lineages. These data show that the fertility of these animals can seriously affect the cultivated stocks, thus causing financial damage in this aquaculture system. The establishment of PCR‐RFLP and multiplex‐PCR as molecular techniques allows for both the correct management of these animals and the routine monitoring of production and trade of fish hybrids in aquaculture. Consequently, such tools will enable a sustainable development in the aquaculture industry.     

Improvement of Matrinxã, Brycon amazonicus,Larviculture by Exposing Eggs to Triiodothyronine

This work evaluated the effect of triiodothyronine (T₃) on larviculture of matrinxã, Brycon amazonicus. Oocytes of three females were pooled, fertilized with pooled semen of two males and separated in four batches that were immersed in triiodothyronine solutions as follows: M₁ (control – water); M₂ (0.01 mg/L T₃); M₃ (0.05 mg/L T₃); and M₄ (0.1 mg/L T₃). Triiodothyronine did not affect fertilization rate and number of hatched larvae. Weight of hatched larvae was significantly higher in treatments M3 and M4, as well as among larvae sampled at Day 12 in all treatments. After 12 d of rearing, biomass gain was higher in the hormone treatments (M₁ 688 ± 569 mg; M₂ 2436 ± 562 mg; M₃ 3572 ± 569 mg; and M₄ 4129 ± 770 mg). In general, coefficients of variation of weight (CV_w) and length (CV_l) did not differ among treatments and cannibalism was registered between 36 and 72 hours post‐hatching (h.p.h.) without differences among treatments. Larval survival increased in the hormone treatments (M₁ 26.5%; M₂ 37.6%; M₃ 40.6%; and M₄ 40.8%). The results indicate that the immersion of matrinxã eggs in triiodothyronine can promote beneficial effects to its larviculture and indicate promising perspectives for culture of this tropical species.     

Polyarthritis associated with visceral leishmaniasis in a juvenile dog

Canine visceral leishmaniasis (CL) is a zoonosis and a chronic systemic disease characterized by a wide spectrum of clinical signs, including, in rare occasions, polyarthritis. This report describes a case of CL in an 8-month old male boxer dog with a history of lameness, fever and lymphadenopathy. A definitive diagnosis of CL was based on the observation of the Leishmania amastigotes seen concomitantly, and for the first time, in the lymph nodes aspiration smears (in macrophages), synovial fluid (in macrophages and neutrophils) and blood (in neutrophils). Despite this extensive dissemination of the parasite, the animal was successfully treated with a multi-step combination of meglumine antimoniate, aminosidine and allopurinol.     

Microbial processes controlling P availability in forest spodosols as affected by soil depth and soil properties

Because carbon dioxide (CO₂) concentration is rising, increases in plant biomass and productivity of terrestrial ecosystems are expected. However, phosphorus (P) unavailability may disable any potential enhanced growth of plants in forest ecosystems. In response to P scarcity under elevated CO₂, trees may mine deeper the soil to take up more nutrients. In this scope, the ability of deep horizons of forest soils to supply available P to the trees has to be evaluated. The main objective of the present study was to quantify the relative contribution of topsoil horizons and deep horizons to P availability through processes governed by the activity of soil micro-organisms. Since soil properties vary with soil depth, one can therefore assume that the role of microbial processes governing P availability differs between soil layers. More specifically, our initial hypothesis was that deeper soil horizons could substantially contribute to total plant available P in forested ecosystems and that such contribution of deep horizons differs among sites (due to contrasting soil properties). To test this hypothesis, we quantified microbial P and mineralization of P in ‘dead’ soil organic matter to a depth of 120 cm in forest soils contrasting in soil organic matter, soil moisture and aluminum (Al) and iron (Fe) oxides. We also quantified microbiological activity and acid phosphomonoesterase activity. Results showed that the role of microbial processes generally decreases with increasing soil depth. However, the relative contribution of surface (litter and 0–30 cm) and deep (30–120 cm) soil layers to the stocks of available P through microbial processes (51–62 kg P ha^?1) are affected by several soil properties, and the contribution of deep soil layers to these stocks vary between sites (from 29 to 59%). This shows that subsoils should be taken into account when studying the microbial processes governing P availability in forest ecosystems. For the studied soils, microbial P and mineralization of P in ‘dead’ soil organic matter particularly depended on soil organic matter content, soil moisture and, to a minor extent, Al oxides. High Al oxide contents in some sites or in deep soil layers probably result in the stabilization of soil organic compounds thus reducing microbiological activity and mineralization rates. The mineralization process in the litter also appeared to be P-limited and depended on the C:P ratio of soil organic matter. Thus, this study highlighted the effects of soil depth and soil properties on the microbial processes governing P availability in the forest spodosols.     

Synthesis and in-vitro antifungal activity of 6-trifluoromethylpyrazolo[3,4-d]pyrimidines

Thirty 6-trifluoromethylpyrazolo[3,4-d]pyrimidines were synthesised and tested for antifungal activity in vitro against ten phytopathogenic fungi of different taxonomic classes. Six of the compounds had noteworthy activity against Sclerotinia minor, Corticium solani and Phoma betae. The relationships between structure and activity are discussed on the basis of the present and previous studies.     

Predictive value of prepartum serum metabolites for incidence of clinical and subclinical mastitis in grazing primiparous Holstein cows

The aim of this study was to measure changes in biochemical markers in the peripartum period of primiparous Holstein cows diagnosed with subclinical and clinical mastitis. In this study, 37 dairy cows were monitored daily during milking until 60 days postpartum and were categorized according to the occurrence of clinical mastitis (group mastitis (GM), n?=?9) or subclinical mastitis (group subclinical mastitis (GSUB), n?=?10) or absence of symptoms (control group (CG), n?=?18). Blood samples were collected weekly from ?30 to 60 days from calving. Samples were grouped for prepartum (?30 to 0 days from calving), early postpartum (0 to 30 days from calving), and late postpartum (30 to 60 days from calving) periods. Prepartum serum non-esterified fatty acid (NEFA) concentration was higher in GM than in CG (P?<?0.01). In addition, CG had higher prepartum serum glucose concentration than GM (P?=?0.03). In the early postpartum period, aspartate aminotransferase (AST) activity was lower in CG than in GSUB (P?<?0.05), and in the late postpartum period, AST activity was lower in CG than GSUB and GM (P?=?0.01). Somatic cell count was higher during the early and late postpartum periods for GM and GSUB when compared to CG (P?<?0.01). In this study, primiparous cows with low glucose and higher NEFA in the prepartum were more susceptible for mastitis in the early postpartum, probably due to low immune function associated to a more negative energy balance. In sum, increased prepartum serum NEFA concentration and decreased glucose in primiparous cows were associated with clinical mastitis incidence in the postpartum period.     

Assessment of the usage of biodegradable polymeric matrix in vaginal devices to sustain progesterone release in cows

The usage of timed artificial insemination (TAI) at a low cost leading to better reproductive rates has been the aim of several research groups in the field. Usually during TAI protocols, sustained progesterone (P₄) release devices are employed. Most devices are constituted of a nylon skeleton covered with a silicon layer with P₄. A device based on biopolymers was developed in order to reduce costs and decrease its environmental impact. In this study, we compared the kinetics of sustained progesterone release among devices manufactured with a polymeric blend made of polyhydroxybutyrate‐valerate (PHBV) and poly‐ε‐caprolactone (PCL) (DISP) which were compared with DIB® (Internal Bovine Device) used as the control. In the in vitro and in vivo progesterone release tests, two types of biopolymer‐based devices with a superficial area of 147 cm² were used: DISP8 (46% PHBV, 46% PCL and 8% P₄; n  = 4), DISP10 (45% PHBV, 45% PCL, 10% P₄; n  = 4) and DIB® (1 g P₄, 120 cm² area; n  = 3). The in vitro tests were carried out according to USP XXIII specifications and were performed in a dissolutor sink using an alcohol/water mixture (60/40 v/v) as a release media and samples were collected at 2 min, 2, 4, 8, 12, 24, 48, 60, 72, 84 and 96 h. P₄ concentrations were measured through spectrophotometry in a 244 nm long wave. Three to 3 comparisons of angular coefficients of the straight lines obtained by regression analysis of accumulated P₄ concentrations as a function of square root of time were carried out. Furthermore, the diffusion coefficient values of P₄ were also determined for DISP8 and DISP10. The results showed that the concentrations of P₄ were higher in the DISP10 (774.63 ± 45.26 μg/cm²/t^1/2) compared to DISP8 (566.17 ± 3.68 μg/cm²/t^1/2) (P  < 0.05). However, both DISP10 and DISP8 P₄ concentrations did not differ from DIB® (677.39 ± 16.13 μg/cm²/t^1/2). For the analysis of released quantities per day of the in vitro test, four periods were considered: 0–24, 24–48, 48–72 and 72–96 h. In the first 24 h, DISP8 released significantly less P₄ than DISP10 or DIB®, which did not differ among them. Between 24 and 48 h, DISP10 released significantly more P₄ than DIB®. DISP8 released an intermediate P₄ amount and did not differ significantly from DIB® or DISP10. Between 48 and 72 h, P₄ quantity released by DISP10 was significant higher (P  < 0.01) than that of DIB® and DISP8, which did not differ among themselves. Between 72 and 96 h, DISP10 released significantly more P₄ than DIB®, and DISP8 released an intermediate amount which did not differ from DIB® or DISP10 (P  < 0.01). There was interaction between treatment and time (P  = 0.0024). The diffusion coefficient values were: 1.36 × 10^?8 (cm²/s) for DISP10 and 1.12 × 10^?8 (cm²/s) for DISP8. For the in vivo test, ovariectomized crossbred cows received DIB® (n  = 4) or DISP8 (n  = 8) in an alternate design with a non‐balanced sequence (cross‐over) added of measures repeated in time referring to 16 days of blood samples collection. Samples were analyzed through radioimmunoassay in solid phase using the commercial kit of DPC (Diagnostics Products Corporation). Plasma concentrations of P₄ peaked at 4 h after the placement of the device, this being the only time in which plasma P₄ concentrations differed between DIB® (11.45 ± 1.96) compared with DISP8 (9.23 ± 1.15 ng/mL) (P  = 0.027). On day 8, plasma P₄ concentrations were similar for DIB® (2.44 ± 0.09) and DISP8 (1.89 ± 0.13 ng/mL) (P  = 0.58) showing that both devices were able to keep P₄ concentrations above 1 ng/mL in the plasma of the cow during the 16 day in vivo test. In conclusion, devices manufactured with the blend of PHBV/PCL biopolymers can sustain the release of P₄ in a similar manner as silicon.