Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.     

Deep‐water cirripedes colonizing dead shells of the cephalopod Nautilus macromphalus from New Caledonian waters

Fossil cephalopods are frequently encrusted by epibionts; however, determining whether encrustation occurred prior to or post‐mortem to the host, and whether the final environment of deposition corresponds to the habitat of encrustation is complex. The present paper describes cirripede epibionts, their calcareous bases and their attachment scars on 6 post‐mortem shells of Nautilus macromphalus, collected from deep water off New Caledonia. The cirripedes have left both cemented calcareous bases of Hexelasma and scars associated with bioerosion and discoloration produced by verrucomorph barnacles. Live cirripedes included a Metaverruca recta, with articulated opercular plates and organic tissue (on a shell that had been exposed on the sea floor for at least 150 years), and specimens of Hexelasma velutinum, one of which was partly attached to an internal surface of a shell. The disposition of verrucomorphs indicates that most Nautilus shells were colonized post‐mortem rather than during a floating stage. However, as cirripedes are known to have colonized living Nautilus, some Hexelasma, preserved only as calcareous eroded bases, may represent specimens that settled on a living Nautilus. The degree of bioerosion and discoloration induced by verrucomorph barnacles varies according to the surface preservation of Nautilus shells, with deeper and discolored traces preserved on old and degraded shells. Traces made by verrucomorphs described here are ellipsoidal and a new ichnotaxon, Anellusichnus ellipticus, is proposed to accommodate them. Importantly, verrucomorphs and other cirripede taxa with membranous bases that were attached to pristine shells may not leave any substantial scars, and, thus, will be difficult to detect in the fossil record.     

Photoperiod and temperature affects seasonal ovarian gene expression of p450 aromatase and gonadotropin receptors in Atlantic Salmon (Salmo salar L.) broodstock

Photoperiod and temperature modulated the seasonal pattern of ovarian gene expression of P450 aromatase, Follicle-Stimulating Hormone receptor and Luteinizing Hormone receptor in Atlantic salmon broodstock.     

Digestive protease activities and free amino acids in white muscle as indicators for feed conversion efficiency and growth rate in Atlantic salmon (Salmo salar L.)

The aim of the present experiment was to screen several biochemical indices in fish and their interrelations in order to select variables for future studies of growth rate and feed conversion. Several parameters [trypsin activity, chymotrypsin activity, free amino acids (FAA) in plasma and white muscle, and RNA and RNA/protein ratio in the white muscle] were measured together with specific growth rate (SGR), feed intake and feed conversion efficiency (FCE) in four groups of diploid or triploid Atlantic salmon (Salmo salar L.) reared under different light regimes. SGR was measured on individually tagged fish, whereas feed intake and feed conversion was estimated on tank basis. A principal component analysis (PCA) explained 80.6% of the variance in the data, using all measured parameters, regardless of ploidy and light regime. Muscle free hydroxyproline showed the highest correlation, alone explaining 55% of SGR variability. The SGR also significantly correlated with trypsin activity (r=0.34), the activity ratio of trypsin to chymotrypsin (T/C) (r=0.39), plasma essential FAA (EAA) (r=0.39), plasma total FAA (TFAA) (r=0.37), the ratio of essential to non-essential FAA (EAA/NEAA) in the white muscle (r=–0.45), muscle RNA (r=–0.45) and RNA/protein ratio (r=–0.41). Tank FCE correlated positively (r=0.97) with SGR, T/C ratio and muscle free hydroxyproline, and negatively (r=–0.90) with muscle EAA/NEAA. The groups reared under continuous light (LL) regime showed significantly higher SGR than simulated natural photoperiod (SNP) groups, and with an apparently higher FCE. A higher growth rate was associated with either a higher consumption rate and/or a higher feed utilization. A negative correlation between muscle RNA concentration and SGR may indicate that increased growth rate under LL regime was not caused by an increased protein deposition rate.     

The combined effects of temperature and GnRHa treatment on the final stages of sexual maturation in Atlantic salmon (Salmo salar L.) females

Atlantic salmon (Salmo salar L.) females (2 SW), maturing for the first time, were reared under one of three temperature regimes (high: 14.3 ± 0.5°C; natural: 10.6 ± 1.0°C; and cold: 6.9 ± 1.0°C) in combination with one of two experimental treatments; an injection of GnRH analogue (GnRHa) contained in biodegradable microspheres, or a sham injection (microspheres only). The six experimental groups were then reared under simulated natural photoperiod for 4 weeks. Blood samples were drawn for analysis of plasma steroid levels and the fish were inspected for ovulation weekly. Batches of stripped eggs were incubated in triplicate incubators in raceways until the eyed stage. Treatment with GnRHa resulted in a substantial advancement and synchronization of ovulation at all temperatures, while exposure to cold water also appeared to advance ovulation slightly. While 75% (warm and cold) to 90% (natural) of GnRHa fish ovulated during the 4-week trial, only 30% of sham-treated females exposed to cold water, and none of the sham-treated fish held at higher temperatures, ovulated during this period. Survival rates of embryos to the eyed-stage were significantly higher for broodstock exposed to cold water. Plasma levels of testosterone (T), 17β-oestradiol (E₂), and 17α,20β-dihydroxy-4-pregnen-3-one (17,20βP) were all significantly affected by treatment with GnRHa and, to a lesser extent, temperature. The efficiency of GnRHa in counteracting the negative effects of high temperature on ovulation and the associated changes in circulating sex steroids suggest that temperature inhibition operates at least in part at the brain or pituitary.     

Comparison of Different Glycerol and Egg Yolk Concentrations Added to Tris‐based Extender for the Collared Peccaries (Tayassu tajacu) Semen Freezing

This study aimed to evaluate various concentrations of egg yolk (5, 10, or 20%) in combination with different concentrations of glycerol (3% or 6%) added to a Tris‐based extender on the post‐thaw characteristics of sperm obtained from Tayassu tajacu. For this purpose, semen from 10 sexually male mature collared peccaries was collected by electroejaculation and evaluated for sperm motility, vigour, viability, morphology and functional membrane integrity. The ejaculates were initially extended in Tris‐fructose plus egg yolk (5%, 10% or 20%). After cooling, the semen was added to Tris‐egg yolk plus glycerol (6% or 12%), resulting in a final concentration of 3% or 6% glycerol of the extender. Straws were frozen using liquid nitrogen and thawed in a water bath at 37°C for 30 s. The frozen–thawed semen was evaluated as reported for fresh semen. After thawing, a significant decrease was verified for sperm motility and vigour, for all the samples in comparison with fresh semen. However, no differences were evidenced among treatments for any sperm characteristics evaluated (p > 0.05), except for the combination between 10% egg yolk and 6% glycerol, which provided the worst preservation of functional membrane integrity (p < 0.05). The interactions between higher concentrations of egg yolk (20%) and glycerol (6%) and also between lower concentrations of the same substances (5% egg yolk and 3% glycerol) added to the Tris‐based extender negatively affected the preservation of the normal sperm morphology after thawing (p < 0.05). In conclusion, the use of Tris‐based extender added to 10% or 20% egg yolk plus 3% glycerol is recommended for effective sperm cryopreservation in collared peccaries.     

Immunological studies on urinary bladder tumors of rats and mice

Human neoplasms derived from the same tissue have been previously shown to have tumor associated antigens characterizing that tissue type. Evidence is now presented for the existence of analogous antigens common to both rat bladder papillomas and carcinomas, and for antigens common to mouse bladder carcinomas. Rats immunized with syngeneic urinary bladder papillomas, then challenged with a methylcholanthrene pellet inserted into the bladder, develop (4 to 6 months later) fewer primary bladder tumors than rats immunized with normal bladder tissue.