Olivier Henriet

Exploring the diversity of extremely halophilic archaea in food-grade salts

Salting is one of the oldest means of food preservation: adding salt decreases water activity and inhibits microbial development. However, salt is also a source of living bacteria and archaea. The occurrence and diversity of viable archaea in this extreme environment were assessed in 26 food-grade salts from worldwide origin by cultivation on four culture media. Additionally, metagenomic analysis of 16S rRNA gene was performed on nine salts. Viable archaea were observed in 14 salts and colony counts reached more than 10(5)CFU per gram in three salts. All archaeal isolates identified by 16S rRNA gene sequencing belonged to the Halobacteriaceae family and were related to 17 distinct genera among which Haloarcula, Halobacterium and Halorubrum were the most represented. High-throughput sequencing generated extremely different profiles for each salt. Four of them contained a single major genus (Halorubrum, Halonotius or Haloarcula) while the others had three or more genera of similar occurrence. The number of distinct genera per salt ranged from 21 to 27. Halorubrum had a significant contribution to the archaeal diversity in seven salts; this correlates with its frequent occurrence in crystallization ponds. On the contrary, Haloquadratum walsbyi, the halophilic archaea most commonly found in solar salterns, was a minor actor of the food-grade salt diversity. Our results indicate that the occurrence and diversity of viable halophilic archaea in salt can be important, while their fate in the gastrointestinal tract after ingestion remains largely unknown.

Influence of feeding pattern and hydraulic selection pressure to control filamentous bulking in biological treatment of dairy wastewaters

In sequencing batch reactors (SBRs) treating dairy wastewaters, the overgrowth of filamentous bacteria is a frequent cause of operational problems. The present study aimed at understanding to what extent the operating conditions of a SBR can be optimized to convert a bulking activated sludge into a well-settling biomass at low aeration velocity. The abundance of filament morphotypes and floc-formers able to store biopolymers were analysed by PCR-DGGE and 16S amplicon sequencing. The results indicated that a combination of an anaerobic-microaerated feeding pattern with a low selective pressure was beneficial to supress filamentous overgrowth and to form aerobic granules, while increasing the efficiency of suspended solid removal. Average removal efficiencies for total chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP) were 94±2%, 95±1% and 83±13%, respectively.

Filamentous bulking caused by Thiothrix species is efficiently controlled in full-scale wastewater treatment plants by implementing a sludge densification strategy

Filamentous bulking caused by Thiothrix species is responsible for sludge washout and loss of performance in dairy wastewater treatment plants. A long-term study was conducted over 1.5 years to test three different mitigation strategies in a full-scale plant composed of two parallel sequential batch reactors (SBR1 and 2). Strategies based on polyaluminium chloride addition and volatile fatty acids reduction were ineffective to permanently solve the problem. On the contrary, modification of the reactor cycle based on the implementation of a periodic starvation proved efficient to solve the biomass wash-out and drastically reduce the sludge volume index in both reactors. Bacterial diversity analysis using 16S amplicon sequencing and quantitative PCR indicated a reduction of Thiothrix abundance from 51.9 to 1.0% in SBR1 and from 71.8 to 0.6% in SBR2. Simultaneously, the abundance of the glycogen-accumulating bacterium Candidatus Competibacter increased in both reactors. Microscopy analysis confirmed the transition between a bulking sludge towards a granular-like sludge. This study confirms the applicability of a periodic starvation to (1) solve recurring Thiothrix bulking, (2) convert loose aggregates into dense and compact granular-like structures and (3) considerably reduce energy demand for aeration.