Pine processionary moth (Thaumetopoea pityocampa) and great tit (Parus major) in Portugal

In the present work, we studied a common outbreaking Lepidoptera species in Portuguese pine stands – Thaumetopoea pityocampa (Den. & Schiff.) - and one of its potential predators – Parus major (L.). The population dynamics of the immature stages of the Lepidoptera was studied in several types of Pinus pinaster (Aiton) plantations in three different areas: Setúbal Peninsula, Abrantes and National Pine Forest of Leiria. Location and plantation structure was the most important factors determining population density of T. pityocampa. Setubal and Abrantes was highly susceptible to attacks by the Lepidoptera, whereas Leiria had lower densities. Young and homogeneous pine stands was more susceptible to attacks than older and more heterogeneous pines stands. However, a desynchronized population of T. pityocampa, in which the larvae develops during summer instead of during winter, reached high densities also in Leiria. The impact of several mortality factors and climatic conditions on the immature stages of the insect (eggs and larvae), in normal and desynchronized populations are discussed, as well as possible evolutionary implications of the sudden appearance of the new version of T. pityocampa. The break of the pupa diapause and adult emergence times the annual life cycle of this insect. Adults from the desynchronized population emerged earlier than adults from the normal population, which in turn determined the change in the larvae development period. Different factors, potentially affecting the timing of adult emergence in both normal and abnormal populations are also discussed. To study P. Major, nest-boxes were placed in the areas of Setúbal and Leiria and they were monitored during three seasons. The nest-boxes increased the density of breeding and wintering birds in the studied pine plantations, indicating that a lack of natural holes are in fact a limiting factor for this populations. The earliest breeding start for this species was recorded in my study area, indicating that Portuguese coastal pines provide good breeding conditions earlier than in other areas of Europe and North Africa. This leads to an overlap between the end of the larvae stage of T. pityocampa and the beginning of the breeding season of P. major. Key-words: Thaumetopoea pityocampa, Parus major, Pinus pinaster, population dynamics, Portugal.

Enrichment of a PHA producing microbial community in a continuous bioreactor setup

Polyhydroxyalkanoates (PHAs) are biosynthetic polyesters, biodegradable and biocompatible making them of great interest for industrial purposes. The use of low value substrates with mixed microbial communities (MMC) is a strategy currently used to decrease the elevated PHA production costs. PHA production process requires an important step for selection and enrichment of PHA-storing microorganisms which is usually carried out in a Sequencing Batch Reactor (SBR). The aim of this study was to optimize the PHA accumulating culture selection stage using a 2-stage Continuous Stirrer Tank Reactor (CSTR) system. The system was composed by two separate feast and famine bioreactors operated continuously, mimicking the feast and famine phases in a SBR system. Acetate was used as carbon source and biomass seed was highly enriched in Plasticicumulans acidivorans obtained from activated sludge. The system was operated under two different sets of conditions (setup 1 and 2), maintaining a system total retention time of 12 hours and an OLR of 2.25 Cmmol/L.h-1. An average PHB-content of 3.3 % wt was obtained in setup 1 and 4.8% wt in setup 2. Several other experiments were performed in order to better understand the continuous system behaviour, using biomass from the continuous system. With the fed-batch experiment a maximum of 8.1% PHB was stored and the maximum substrate uptake and specific growth rates obtained in the growth experiment (1.15 Cmol Cmol-1.h-1 and 0.53 Cmol Cmol-1.h-1) were close to the ones from continuous system (1.12 Cmol Cmol-1.h-1 and 0.59 Cmol Cmol-1.h-1). The microbial community was characterized trough microscopic visualization, Denaturing Gradient Gel Electrophoresis (DGGE) analysis and Fluorescent in situ hybridization (FISH). The last studied performed mimicked the continuous system by building up a SBR system with all the same operational conditions while adding an extra acetate dosage during the 12 h cycle, simulating the substrate passing from the feast to the famine reactors under continuous operation. It was shown that possibly the continuous system was not able to efficiently select for PHB storing organisms under the operational conditions imposed, although the selected culture was capable of consuming the substrate and grow fast. This main conclusion might have resulted from two major factors affecting the system performance: the ammonium concentration in the Feast reactor and the amount of substrate leaching from the Feast to the Famine reactor.