Although increasing attention has been attracted to the study and application of biofloc technology (BFT) in aquaculture, few details have been reported about the bacterial community of biofloc and its manipulation strategy for commercial shrimp production. An 8-week trial was conducted to investigate the effects of three input C/N ratios (8:1, 12:1 and 16:1) on the bacterial community of water biofloc and shrimp gut in a commercial BFT tank system with intensive aquaculture of P. vannamei. Each C/N ratio group had three randomly assigned replicate tanks (culture water volume of 30 m³), and each tank was stocked with juvenile shrimp at a density of 300 shrimp m⁻³. The tank systems were operated with zero-water exchange, pH maintenance and biofloc control. During the trial, the microbial biomass and bacterial density of water biofloc showed similar variation trends, with no significant difference under respective biofloc control measures for the three C/N ratio groups. Significant changes were found in the alpha diversity, composition and relative abundance of bacterial communities across the stages of the trial, and they showed differences in water biofloc and shrimp gut among the three C/N ratio groups. Meanwhile, high similarity could be found in the composition of the bacterial community between water biofloc and shrimp gut. Additionally, nitrogen dynamics in culture water showed some differences while shrimp performance showed no significant difference among the three C/N ratio groups. Together, these results confirm that the manipulation of input C/N ratio could affect the bacterial community of both water biofloc and shrimp gut in the environment of a commercial BFT system with intensive production of P. vannamei. Moreover, there should be different operations for the nitrogen dynamics and biofloc management during shrimp production process under different C/N ratios.

Ammonia is the main pollution factor of the aquatic environment in marine shrimp culture systems. In order to demonstrate the feasibility of the combination of biofloc technology and nitrifying biofilter for the ammonia removal, a 70-day production trial was conducted in a simplified pilot-scale hybrid biofloc-based recirculating aquaculture system (biofloc-RAS) with the intensive culture of Litopenaeus vannamei. Nitrogen dynamics and nitrifying microbial communities were investigated in three replicated systems simultaneously under the conditions of high feed loading and zero water exchange. Along with biofloc development in the culture tank and biofilm formation in the nitrifying biofilter during the trial, nitrification could be fastly and effectively established in the system, which was indicated by the dynamics of total ammonia nitrogen (TAN), NO₂^–-N, NO₃^–-N, and total nitrogen (TN) concentrations. Meanwhile, similar nitrifying microorganisms could be found between biofloc and biofilm, despite some differences in abundance, diversity, and composition of ammonia-oxidizing archaea and bacteria and nitrite-oxidizing bacteria. High TAN removal rate could be achieved and was significantly and positively correlated with abundances of these nitrifying microbial communities in both biofloc and biofilm, further indicating that both biofloc and biofilm could contribute highly to nitrification performance of the biofloc-RAS. The results of this study indicate a potential application of the biofloc-RAS in coastal intensive aquaculture.

Frequent harmful cyanobacteria blooms limit the sustainable development of aquaculture. Algicidal bacteria can efficiently control harmful algae without secondary pollution. The algicidal bacteria CZBC1 can lyse Oscillatoria spp. and other harmful cyanobacteria, but its effector mechanism and algicidal threshold are unknown. In this study, we examined the algicidal effect of CZBC1 on O. chlorina, O. tenuis, and O. planctonica by microscopic enumeration and scanning electron microscopy observation. Then, we examined the alginolytic effects of CZBC1 (concentrations 10³–10⁶ colony forming units (cfu)/mL) on these three species (concentrations 10³–10⁶ cells/mL) to determine the effective concentrations of CZBC1 for Oscillatoria spp. alginolysis. Results showed that CZBC1 can directly lyse O. chlorina and O. tenuis but indirectly lyse O. planctonica. When the initial concentration of CZBC1 was 10⁶ cfu/mL, alginolytic effects were high for all three species at all concentrations, and the alginolytic rate could reach 100% in 3–9 days. When the initial concentration of CZBC1 was lower (10³ cfu/mL), its inhibitory effects were delayed by 2–5 days, but the cell counts were significantly decreased compared with the control, evidencing significant alginolysis. In addition, the higher the concentration of the algicidal bacteria suspension, the more significant the alginolytic effects. Our results indicate that CZBC1 has different alginolytic mechanisms for O. chlorina, O. tenuis, and O. planctonica, and that different initial concentrations of CZBC1 have different alginolytic effects on these algal species.

In this study, we developed a rapid and effective method for enriching the culture of nitrifying bioflocs (NBF) from aquacultural brackish water. The self-designed mixotrophic mediums with a single or mixed addition of sodium acetate, sodium citrate, and sucrose were used to investigate the enrichment process and nitrification efficiency of NBF in small-scale reactors. The results showed that NBF with an MLVSSs from 1170.4 mg L⁻¹ to 2588.0 mg L⁻¹ were successfully enriched in a period of less than 16 days. The citrate group performed the fastest enrichment time of 10 days, while the sucrose group had the highest biomass of 2588.0 ± 384.7 mg L⁻¹. In situ testing showed that the highest nitrification efficiency was achieved in the citrate group, with an ammonia oxidation rate of 1.45 ± 0.34 mg N L⁻¹ h⁻¹, a net nitrification rate of 2.02 ± 0.20 mg N L⁻¹ h⁻¹, and a specific nitrification rate of 0.72 ± 0.14 mg N g⁻¹ h⁻¹. Metagenomic sequencing revealed that Nitrosomonas (0.0~1.0%) and Nitrobacter (10.1~26.5%) were dominant genera for AOB and NOB, respectively, both of which had the highest relative abundances in the citrate group. Linear regression analysis further demonstrated significantly positive linear relations between nitrification efficiencies and nitrifying bacterial genera and gene abundance in NBF. The results of this study provide an efficient enrichment culture method of NBF for the operation of biofloc technology aquaculture systems, which will further promote its wide application in modern intensive aquaculture.

Given the widespread use of commercial probiotics in aquaculture, it is important to evaluate the quality and environmental effects of these additives. Here, the effects of a Bacillus probiotic compound, BG4, on Penaeus vannamei survival rate, water quality factors, and microbial communities were assessed. An analysis of the BG4 powder confirmed the content of probiotic bacteria: 50.2% Bacillus licheniformis, 48.4% Bacillus subtilis, and 1.4% Bacillus amyloliquefaciens, consistent with the information on the product packaging. The effective bacterial quantity (1.1 × 10⁹ CFU/g) was higher than that indicated on the product label (2 × 10⁸ CFU/g). BG4 was added to a sterilized aquaculture water system, according to the indicated bacterial quantity; after 7 days, the maximum ammonia nitrogen, phosphate, and chemical oxygen demand degradation rates were 36.3%, 28.9%, and 15.2%, respectively. In the shrimp culture experiment, the survival rate of P. vannamei and water quality did not differ significantly between the BG4 and control groups. The bacterial quantity and high-throughput sequencing results indicated that Bacillus content in BG4 group decreased continuously from 3.5 × 10⁴ CFU/mL to 6.9 × 10² CFU/mL. To ensure the desired effect of probiotics in commercial aquaculture applications, additional quality evaluations and scientific assessments are needed.

In this study, an eight-week culture trial of Penaeus vannamei juveniles was conducted in commercial intensive systems to compare the impacts of biochar and molasses addition on biofloc nitrifying bacteria and inorganic nitrogen dynamics under limited water exchange conditions. During the trial, the biofloc concentration (in terms of VSS and TSS), quantities of total bacteria (TB) and total Vibrio (TV), and ratio of TV/TB in the culture water were lower in the biochar group compared to the molasses group. Metagenomic sequencing analysis revealed that the bacterial community composition of bioflocs showed higher α-diversity and complexity in the biochar group compared to the molasses group. Moreover, the abundance of nitrifying bacterial genera and functional genes in bioflocs was higher in the biochar group compared to the molasses group. Inorganic nitrogen dynamics showed that NH₄⁺-N and NO₂⁻-N were better controlled in the biochar group compared to the molasses group, as reflected by lower peaks of NH₄⁺-N and NO₂⁻-N and higher NO₃⁻-N concentrations. Excellent production performance of shrimp was achieved, which in turn proved the reliable effect of biochar addition on the mediation of inorganic nitrogen transformation through nitrifying bacteria. These results showed that biochar addition could promote biofloc nitrifying bacteria and nitrification to more effectively control harmful nitrogen for shrimp efficient production. This study provides a practical example for the biochar application in biofloc-based systems for intensive aquaculture.