Effect of thermal treatment on free-living amoeba inactivation.

AIMS: To evaluate the effect of temperature on two amoeba strains of the genera Acanthamoeba and two amoeba strains of the genera Hartmannella separately treated depending on their life stage, trophozoite or cyst, when cells are directly exposed under controlled conditions. METHODS AND RESULTS: For thermal treatments, three temperatures were selected 50, 60 and 70°C, and a microcosm was designed using dialysis bags. The inactivation of each strain was determined using a method based on the most probable number quantification on agar plates. The results showed that for all amoeba strains, thermal treatment was more effective against trophozoites compared with cyst stages. The inactivation patterns showed statistical differences between the two genera analysed at temperatures above 50°C. The effectiveness of the thermal treatments at 60 and 70°C was higher for both life stages of Hartmannella vermiformis strains compared with Acanthamoeba strains, being the most resistant Acanthamoeba cysts. CONCLUSIONS: Free-living amoebae have been isolated in a wide range of environments worldwide due to their capacity to survive under harsh conditions. This capacity is mainly based on the formation of resistant forms, such as double-walled cysts, which confers a high level of resistance as shown here for thermal treatments. SIGNIFICANCE AND IMPACT OF STUDY: Free-living amoebae survival can promote a rapid recolonization of drinking water systems and is a likely source of emerging opportunistic pathogens such as Legionella. Because of that a better understanding of the factors that affect micro-organism inactivation in water systems would allow more efficient application of disinfection treatments.

Microfauna communities as performance indicators for an A/O Shortcut Biological Nitrogen Removal moving-bed biofilm reactor.

The microfauna communities present in the mixed liquor and biofilm of an Anoxic/Oxic Shortcut Biofilm Nitrogen Removal moving-bed biofilm process were characterised in order to optimise process control through the use of bioindicators. The system operated at high ammonium concentrations, with an average of 588 ± 220 mg N-NH4(+) L(-1) in the influent, 161 ± 80 mg L(-1) in the anoxic reactor and 74 ± 71.2 mg L(-1) in the aerobic reactor. Up to 20 different taxa were identified, including ciliates (4), flagellates (11), amoebae (4) and nematodes (1). Compared to conventional wastewater treatment processes (WWTPs), this process can be defined as a flagellates-predominant system with a low diversity of ciliates. Flagellates were mainly dominant in the mixed liquor, demonstrating high tolerance to ammonium and the capacity for survival over a long time under anoxic conditions. The data obtained provide interesting values of maximum and minimum tolerance ranges to ammonium, nitrates and nitrites for the ciliate species Cyclidium glaucoma, Colpoda ecaudata, Vorticella microstoma-complex and Epistylis cf. rotans. The last of these was the only ciliate species that presented a constant and abundant population, almost exclusively in the aerobic biofilm. Epistylis cf. rotans dynamics showed a high negative correlation with ammonium variations and a positive correlation with ammonium removal efficiency. Hence, the results indicate that Epistylis cf. rotans is a good bioindicator of the nitrification process in this system. The study of protozoan communities in unexplored WWTPs sheds light on species ecology and their role under conditions that have been little studied in WWTPs, and could offer new biological management tools.

Tissue inhibitor of metalloproteinases-1 (TIMP-1) is differentially induced in neurons and astrocytes after seizures: evidence for developmental, immediate early gene, and lesion response.

We investigated in vivo the expression of the tissue inhibitor of metalloproteinases-1 (TIMP-1) in the rat CNS after kainate (KA)-induced excitotoxic seizures. In situ hybridization revealed that TIMP-1 mRNA is induced rapidly and massively in most regions of the adult forebrain after KA treatment. Neuronal activity seems to be necessary but not sufficient to trigger TIMP-1 induction, because it is not observed in seizing 10-d-old pups, unlike what is observed in 21- and 35-d-old animals after seizures. The rapid induction of TIMP-1 is not prevented by the inhibitor of protein synthesis cycloheximide, suggesting that, after seizures, TIMP-1 is induced in neurons as an immediate early gene (IEG). The initial neuronal upregulation is followed by enhanced expression in astrocytes, as assessed by double-labeling experiments. In the hippocampus rapid increases in mRNA are followed by relatively delayed (8 hr after KA) increases in TIMP-1 immunoreactivity in the perisomatic and dendro-axonic areas, suggesting secretion of the protein. At 3 d after KA treatment, strong immunoreactivity is found in astrocytes and in the cell bodies and dendro-axonic projections of resistant neurons such as the dentate granule cells. Taken together, the results suggest that TIMP-1 may be instrumental for neurons and astrocytes in coupling early cellular events triggered by seizures with the regulation of long-lasting changes involved in tissue reorganization and/or neuroprotection.