This study examined the effects of Tribulus terrestris (TT) on sex reversal in guppy, Poecilia reticulata. The objective of this study was to introduce a new environmentally friendly method for masculinization in P. reticulata. Since male guppy has higher commercial value than female. TT is a natural, non-toxic herb which helps enhance testosterone levels in human and animals. It was prepared in a laboratory in France. Different concentration (0.0, 0.05, 0.1 and 0.15 g L(-1)) of TT was investigated for sex reversal in the Poecilia reticulata. TT extract was administered by immersion of newly born offspring once weekly for two months. Among the dosages used in the present study 0.15 g L(-1) TT was the most effective dosage that ensured maximum male ratio (80%, p < 0.01). Although, sex ratios of 0.05 and 0.1 g L(-1) TT were not significantly different from the expected 1:1 ratio, in these two groups treatment with TT also result in higher number of males (58.25 and 59.77%, respectively), than control (p > 0.05). Total survival rates in all treatments and control were uniformly high ranging from 83 to 87% (p > 0.05). It is concluded that TT has no negative effect on survival rate of P. reticulata. All groups of TT-treated fish exhibited successful growth acceleration comparing to the control group, but only TT treatment at the concentration of 0.15 and 0.1 g L(-1) TT significantly improved growth rate of P. reticulata (p < 0.01). Histological examinations revealed that testes of fish treated with TT-extract contained all stages of spermatogenesis. Sex reversal in P. reticulata demonstrated that TT treated new-born progenies showed successful sex reversal, spermatogenesis and better growth rate than untreated progenies.

We have examined the effects of Tribulus terrestris (TT), a non-toxic herb, on sex reversal in the Convict Cichlid Cichlasoma nigrofasciatum with the aim of introducing a new environmentally friendly method for masculinization in C. nigrofasciatum. TT is a natural plant product that elevates the testosterone levels in humans and animals. Different concentrations (0.0, 0.10, 0.20 and 0.30 g/l) of TT extract were tested for their effect on sex reversal in C. nigrofasciatum by immersing newly hatched offspring once weekly for 2 months in TT extract. Of the dosages used in the present study, 0.30 g/l TT was the most effective in terms of masculinization, resulting in a maximum male ratio of 87.23% (P < 0.001). Sex ratios of 79 and 85% at 0.10 and 0.20 g/l TT, respectively, were also significantly different from the expected 1:1 ratio (P < 0.001). Histological examinations revealed that the testes of fish treated with TT extract contained all stages of spermatogenesis, clearly demonstrating that the administration of TT extract to C. nigrofasciatum stimulated spermatogenesis. Total survival rates in all treatments and the control were uniformly high, ranging from 88.57 to 90% (P > 0.05). We concluded that TT had no negative effect on the survival rate of C. nigrofasciatum. In addition, all groups of TT-treated fish exhibited successful growth acceleration compared to the control group, but only the 0.30 g/l TT treatment significantly improved the growth rate of C. nigrofasciatum. (P < 0.01). Sex reversal in C. nigrofasciatum demonstrated that TT-treated 0-day-old larvae showed successful sex reversal, spermatogenesis and a better growth rate than untreated progenies.

The main objective of this study was to describe the early gonadal development and to examine the process of sex differentiation in male and female P. conchonius under laboratory conditions. First evidence of primordial germ cells was observed on the day of hatching. The sex differentiation in leptotene, zygotene, pachytene and diplotene stages was clearly detected. Differentiation started from the mid-mid posterior part of the gonads. Actual sex differentiation occurred between 18-21 days and 36-40 days post-hatching in females and males, respectively. Histological sex differentiation differences were clear between males and females; in males, gonads had a smooth surface, were less stained, arrow-shaped, with germ cells located alone in the stroma, and number of germ cells ranging from two to ten per section, whereas in females, gonads had a rough surface, were more stained, wider, with germ cells multiplying rapidly and forming clusters, and number of germ cells ranged from 2 to 58 per section. The numbers of germ cells within the two gonad types were significantly different in favour of females (P < 0.05). Here early developmental stages of the gonads from 0 to 56 days post-hatch are explained.

Testicular germ cells of channel catfish, Ictalurus punctatus, and blue catfish, I. furcatus were separated into four layers with Percoll density gradient centrifugation, containing different cell types (40% in the first layer were spermatogonial stem cells, SSCs). Expression of seventeen genes was analyzed for cells from different layers by real-time quantitative PCR. Pfkfb4, Urod, Plzf, Integrin6, IntegrinV, Thy1 and Cdh1 genes showed the same expression change pattern in both channel and blue catfish as these genes were down-regulated in the spermatocytes and even more so in spermatids. Plzf and Integrin6 had especially high expression in SSCs and can be used as SSCs specific markers. Sox2 gene was up-regulated in spermatocytes and even more highly up-regulated in spermatids, which indicated it could be a spermatid marker. In contrast to channel catfish, Id4, Smad5 and Prdm14 gene expressions were strongly down-regulated in spermatocyte cells, but up-regulated in spermatid cells in blue catfish. Smad5 gene was down-regulated in spermatocytes, but up-regulated in both spermatogonia and spermatids, allowing identification as a marker for spermatocytes in blue catfish. Oct4, Id4, Gfrα2, Pum2 and Prdm14 genes showed different expression patterns in the testicular germ cells of channel and blue catfish. This may be a partial explanation to the differing responses of channel catfish and blue catfish to induced spawning technologies. The SSCs specific markers can be used for further SSCs labeling, which can increase the SSCs sorting efficiency and be applied in various studies involving SSCs and other germ cells.