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Giant freshwater prawns (Macrobrachium rosenbergii) are commonly found throughout the world. The size of the male giant freshwater prawn is much larger than that of the female. Therefore, understanding the molecular mechanism that underlies the sexual differentiation of M. rosenbergii is of both commercial and scientific importance. Insulin-like androgenic gland hormone (IAG) plays a key role in the differentiation of sex in M. rosenbergii. Although IAG has been investigated, the regulatory relationship between IAG and its binding protein partner, the insulin-like androgenic gland hormone-binding protein (IAGBP), has not been studied in M. rosenbergii. Here, we cloned and characterized the IAGBP from M. rosenbergii (Mr-IAGBP) for the very first time. Transcriptomic analysis showed that Mr-IAGBP mRNA was detected in a wide array of tissues with the highest expression found in the androgenic gland. The importance of IAG in male development was further demonstrated by an increase in IAG transcripts during the development of the androgenic gland and Mr-IAG was only highly transcribed in the androgenic gland of M. rosenbergii. Interestingly, we found that the Mr-IAG gene expression started during the 20th-day larva after hatching stage (LH20), followed (20th-day post-larval stage, PL20) by a gradual elevation of Mr-IAGBP levels. The levels of both genes peaked at the adult stage. The relationship between Mr-IAGBP and Mr-IAG was further analyzed using RNA interference. The injection of Mr-IAGBP double-stranded RNA (dsRNA) significantly reduced the transcription of Mr-IAG, while the amount of Mr-IAGBP mRNA and the translation of IAGBP protein was significantly reduced by the injection of Mr-IAG dsRNA. These results revealed that IAGBP is involved in IAG signaling. Furthermore, our data supports the hypothesis that (IAG and IAGBP)-IAG receptor signaling schemes exist in M. rosenbergii. Our results will provide important information for the further study of determining the sex of M. rosenbergii.

Haemulidae species are morphologically diversified fishes with wondrous and changeable coloration. These species are prevalent in global tropical, subtropical, and temperate inshore reef areas. Previous morphological classification within Haemulidae and some related families was problematic, and no comprehensive molecular evaluation was conducted on these groups. In this study, we revealed the first molecular phylogenetic analysis involving representative species within Haemulidae and the relevant families (Nemipteridae and Teraponidae). The analysis was performed using both mitochondrial DNA (16S rRNA) and nuclear DNA (TMO-4c4) genes to construct maximum parsimony, maximum likelihood, and Bayesian analysis tree topologies. The molecular phylogeny recovered well-resolved relationships within Haemulidae species and the problematic taxa. In the trees, the Haemulidae species (except for genus Hapalogenys) were divided into two distinct sister lineages, including grunts (Haemulinae) and sweetlips (Plectorhynchinae). Hapalogenys was positioned outside the major Haemulidae tribe and formed an independent group, which challenged the traditional taxonomy that Hapalogenys was classified into Haemulidae. The results did agree with most current studies that Hapalogenys was only distantly related to Haemulidae and could potentially be removed from this family. Additionally, the genus Diagramma was observed to be tightly grouped inside the clade Plectorhinchus, indicating its highly close affinity within Plectorhinchus. Regarding interfamily relationships, the phylogenetic constructions suggested distant relationships within two pairs: between Scolopsis and Haemulidae and between Teraponidae and Haemulidae. Scolopsis was revealed to be highly close with Nemipteridae, and Teraponidae was clustered in an independent group, which was in accordance with most current taxonomic studies.

The aim of this study was to investigate effects of dietary feed additives on growth and immunological status of Litopenaeus vannamei under cold exposure. Four experimental diets with 0.2% taurine, 0.01% carnitine, 0.1% cholesterol supplementation and control were fed to L. vannamei (IBW 0.86 AE 0.03 g) for 30 days at 30 AE 1°C before water temperature was dropped to 16°C with a rate of À4°C h À1 . Growth performance parameters including weight gain rate (WGR), feed conversion ratio (FCR) and survival rate before the cold exposure, as well as immunological parameters related to haemocytes including total haemocyte count (THC), percentages of hyaline cells (SC%), granular cells and semigranular cells (SC + GC%), non-specific esterase (NSE) activity and apoptotic cell ratio before and after the cold exposure were determined. The results showed that: (1) WGR, FCE and survival rate were not significantly different among groups before the cold exposure. Most of immunological parameters did not differ either, except the apoptotic cell ratio of carnitine and cholesterol groups was significantly lower than that of control and taurine groups (P < 0.05). (2) After the cold exposure, THC was decreased by 48.1% (P < 0.05), 54.1% (P < 0.05), 30.1% and 2.2%; NSE activity was decreased by 31.7% (P < 0.05), 21.4%, 7.0% and 3.5% in control, taurine, carnitine and cholesterol respectively. (3) Conversely, ROS production was increased by 23.8% (P < 0.05), 15.1%, 7.2% and 13.0%; apoptotic cell ratio was increased by 10.4%, 12.8%, 6.4% and 7.2% in control, taurine, carnitine and cholesterol respectively. (4) An increase in HC %, decrease in SC + GC% in all groups was observed after cold exposure, and was significant in cholesterol group. This study showed that cholesterol, carnitine and taurine affected immunological status of L. vannamei under cold exposure to different degrees, possibly by modulating some immunological parameters, as discussed. Present study provides theoretical data for nutritional strategies to enhance cold resistance of shrimp.

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