Microbiome of Pacific Whiteleg shrimp reveals differential bacterial community composition between Wild, Aquacultured and AHPND/EMS outbreak conditions.

Crustaceans form the second largest subphylum on Earth, which includes Litopeneaus vannamei (Pacific whiteleg shrimp), one of the most cultured shrimp worldwide. Despite efforts to study the shrimp microbiota, little is known about it from shrimp obtained from the open sea and the role that aquaculture plays in microbiota remodeling. Here, the microbiota from the hepatopancreas and intestine of wild type (wt) and aquacultured whiteleg shrimp and pond sediment from hatcheries were characterized using sequencing of seven hypervariable regions of the 16S rRNA gene. Cultured shrimp with AHPND/EMS disease symptoms were also included. We found that (i) microbiota and their predicted metagenomic functions were different between wt and cultured shrimp; (ii) independent of the shrimp source, the microbiota of the hepatopancreas and intestine was different; (iii) the microbial diversity between the sediment and intestines of cultured shrimp was similar; and (iv) associated to an early development of AHPND/EMS disease, we found changes in the microbiome and the appearance of disease-specific bacteria. Notably, under cultured conditions, we identified bacterial taxa enriched in healthy shrimp, such as Faecalibacterium prausnitzii and Pantoea agglomerans, and communities enriched in diseased shrimp, such as Aeromonas taiwanensis, Simiduia agarivorans and Photobacterium angustum.

Expression analysis of heat shock protein 20 and rhoptry-associated protein 1a in sexual stages and kinetes of Babesia bigemina.

Heat shock protein 20 (HSP-20) and rhoptry-associated protein 1a (RAP-1a) are two proteins considered as candidates to be included in vaccines or diagnostics methods for the control of bovine babesiosis. It has been hypothesized that both genes have a basic function in the cellular physiology of erythrocyte-infecting stages; it is not known if they have a functional role in tick stages. The objective of this work was to analyze whether hsp-20 and rap-1a are expressed in sexual stages and kinetes of Babesia bigemina. Purified RNA from sexual stages and kinetes was analyzed by reverse transcriptase (RT)-PCR with specific primers for hsp-20 or rap-1a. Expression analysis was carried out using an indirect immunofluorescence test with specific antibodies against HSP-20 and RAP-1a. Results obtained by RT-PCR showed amplicons for hsp-20 and rap-1a in sexual stages and kinetes. Positive signals were also detected when sexual stages and kinetes were analyzed with specific antibodies for HSP-20 and RAP-1a. The results obtained here confirm the hypothesis that the genes hsp-20 and rap-1a from B. bigemina are expressed in sexual stages and kinetes and stress the importance of these proteins in the cellular physiology of tick stages.