BIOLOGICAL CONTROL OF PESTS IN BOTANIC GARDEN “GIARDINO DEI SEMPLICI” OF FLORENCE

ALTIERI G.1_{,BIANCHI M.}2_{,CAPACCI A.}2_{,CLAUSER M.}2_{,FABIANI L.}2_{,FERLI S.}2_{,GASPARRINI G.}2_{,GRIGIONI A.}2_{,LANDI M.}2_,

MARRACCINI C.2_{,VALGIMIGLI F.}2_{,VARRIALE S.}2

1_{Istituto Agrario Todi (PG); Studio AGERNOVA- Servizi per l’Agroecologia e la Ricerca} 2_{Orto botanico ‘Giardino dei Semplici’, Museo di Storia Naturale, Università di Firenze}

The presence in the human body of different synthetic chemical residues increases the negative effects that are “aggravating contributory causes” of health risk especially in populations already exposed to other forms of environmental pollution. So the Botanical Garden of Florence, which is located in the city center, began the conversion of cropping system to the biological control of pests to reduce and possibily eliminate the chemical control. The project is very complex: it began with two training courses for the staff of the italian botanical gardens during which the most advanced biological control techiniques were illustrated. The second step was the activation of an agreement with Agernova – Servizi per l’Agroecologia e la Ricerca - to identify pests and organize a series of launches of antagonists and interventions with natural pesticides. Then we monitored parasites in the greenhouses and in the garden, with visual inspections and sticky traps, highlighting a heavy presence of cochineal, mites, thrips and aphids (the latter already parasitized by naturally present wasps), in addition to the inoculation of powdery mildew and black rot and other plant diseases (Botrytis, etc.). The project started in february 2016 by a spray with a compost of Neem oil and potassium soap to reduce the initial population of mealy (Planococcus spp.

Pseudococcus longispinus), blocking the suit young nymphs and washing the sooty mold. On mealybug

outbreaks were introduced larvae Cryptolaemus, a very active predator Coccinellide, with a first launch of “inundative” type, added to the parasitoid Anagyrus pseudococcidi, active on Planococcus spp. Against mites and thrips it was launched the Multipla Preventive inoculation method, with small doses of phytoseiid predatory mites. Roses, suffering from cancer control, were treated with innovative cupric to lingering absorption, cupric with zinc and iron in order to promote scarring and copper and sulfur, to prevent Oidi and Black spot. In the following months were repeated the Cryptolaemus launches localized on the scale insect outbreaks, particularly Pseudococcus longispinus. For red spider mite we introduced predators of Phytoseiulus persimilis species, very fast in the activity and in the reproduction. Three months later the cochineal considerably diminished; at this point the pest control involves the launch of Anagyrus

fusciventris specific parasitoid, to counter the species Pseudococcus longispinus, by a treatment with

Azadirachtin and oil-soap mixture to reduce possible outbreaks, plus the introduction of spider beetles of

Nephus includens species. As regards the thrips, at present there are no major outbreaks therefore it

continues the launches with Amblisejus cucumeris which preys thrips and red spider mite and Ambliseus

swirskii which preys thrips and the rare white fly, the latter now monitored with yellow sticky traps and

towards which there was a first launch of Encarsia formosa and Eretmocerus eremicus.

Results and conclusions: the experiment is giving good results and we try to achive the ambitious goal in collecting data and informations for the scientific community of reference; in fact, in the botanical gardens has increased the sensibility to the use of alternative systems to chemistry for the control of pathogens; but apart from the Giardini botranici Hanbury where since many years they are making a biological control against several diseases and few other situations (such as the biological control of the red palm weevil in the Orto botanico of Catania) there are not many Botanical Gardens which have adopted a rigorous program of biological control for plant diseases.

In addition to sensitizing the staff working in Botanical Gardens, we also aim to motivate the staff who is in charge of public green spaces to pursue the reduction if not he completely elimination of chemical croppings used to contrast plant diseases in urban contests.

At the same time the Botanical Garden of Florence is testing regenerative organic growing techniques with very important results in strengthening plants health. This will also let us eliminate rameic products in the future, for the toxicological studies indicate them as dangerous for the human health, though it is allowed in biological control.

3.6 = PHOSPHORUS AND HEAVY METALS BIOREMEDIATION COMBINED WITH LIPID PRODUCTION BY THE GREEN MICROALGA DESMODESMUS SP.: AN INTEGRATED APPROACH

BRUNO LAURA1_{, RUGNINI LORENZA}1_{, CONGESTRI ROBERTA}1_{, GARREFFA LORENZO}1_{, CIOTOLA VALERIO}1_,

ANTONAROLISIMONETTA2_,GIULIACOSTA3

1_{LBA-Laboratory of Biology of Algae, Department of Biology, via Cracovia 1;} 2_{Department of Chemical}

Sciences and Technologies, Via della Ricerca scientifica 1; 3_{Laboratory of Environmental Engineering,}

Department of Civil Engineering and Computer Science Engineering, via del Politecnico 1; University of Rome “Tor Vergata”, Rome, Italy

Widespread applications of microalgal-based systems in wastewater treatment (WWT) have been proposed since the 50s [1]_{. Anthropogenically derived wastewater can significantly increase the nutrient}

and heavy metal water load posing hazards to the ecosystem and human health [2]_{. The combination of}

algal growth with WWT is a green biotechnology that cuts down the cost of mass production of microalgae and potentially increases the environmental and economic sustainability of WWT process. WWT facilities can exploit microalgae’s ability to reduce the concentration of excess nutrients, such as dissolved nitrogen and phosphorus, as well as heavy metals in wastewater treatment effluents, which is important in view of guaranteeing the good chemical status of the receiving water body (EU Water Framework Directive 2000/60/EC), but also biomass to harvest and process for the bio-product market [3]_.

In this scenario, we isolated a strain of Desmodesmus sp. (VRUC281) from the secondary tank of a municipal wastewater treatment plant. We evaluated its efficiency in removing phosphorus (P), copper (Cu) and nickel (Ni) from the liquid growth medium. After 4 days, total P was lower than 2 mg L-1 _complying

with the European limits for effluent discharge into water bodies, and in 10 days over 90% of P was removed from the liquid medium that contained up to 4.5 mg P L-1_{. In addition, the strain was incubated}

for 48 hours in a mixed solution containing 9.8 and 7.4 mg L-1 _{Cu and Ni, respectively. After exposure, only}

20% of the initial Cu and 60% of the initial Ni were measured in the liquid solutions by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) analysis. Heavy metal adsorption tests were also conducted by using inactivated biomass of the green microalga S. obliquus (strain VRUC280) to analyse the biosorption of copper (Cu) and nickel (Ni). S. obliquus (0.5 gDW) was incubated in 5.0 and 3.9 mg L-1

Cu and Ni solutions, respectively (contact time 0-120 min) and Cu and Ni concentrations were analysed in the solution and also in the microalgae after ultrasound assisted acid extraction [4] _{After 15 minutes, up to}

51% of the initial Cu (407 mg Cu kgDW-1_{) and 47% of Ni (349 mg Ni kgDW}-1_{) were passively adsorbed on the}

cell surface.

The biomass obtained was harvested by sedimentation and centrifugation and used for FAME (fatty acids methyl esters) extraction to evaluate the effects of the presence of heavy metals on lipid accumulation. Our results encourage the research in this field for an efficient and less expensive water clean-up process, by integrating phosphorus and heavy metal removal using microalgae with biomass production, which can be used for biodiesel extraction. This technology can offer an alternative solution to problems of environmental pollution and to the traditional power generation based on fossil fuels, which are generally considered to be unsustainable in the long term.

1) W.J. Oswald, H.B. Gotaas, (1957). Photosynthesis in sewage treatment. Transaction of the American Society of Civil Engineers 122, 73-105.

2)K.S. Kumar, H.U. Dahms, E.J. Won, J.S. Lee, K.H. Shin, (2015). Microalgae- A promising tool for heavy metal remediation. Ecotoxicology and Environmental Safety 113, 329-352.

3) _{M. Kesaano, R.C. Sims, (2014). Algal biofilm based technology for wastewater treatment. Algal Research} 5, 231-240.

4) W.P.C. dos Santos, V. Hatje, D. Santil, A.P. Fernandes, M.G.A. Korn, de M.M. Souza, (2010). Optimization of a centrifugation and ultrasound-assisted procedure for the determination of trace and major elements in marine invertebrates by ICP OES. Microchemical Journal 95, 169-173.

3.6.= IN VITRO SCREENING EVALUATION OF ABSORPTION CAPACITY, TOLERANCE AND TRANSLOCATION

TO CADMIUM IN AUTOTROPHIC POPULUS ALBA CLONES “VILLAFRANCA” MARZILLI M., B.PAURA, COCOZZA C., DI SANTO P., PALUMBO G.

Dipartimento Agricoltura, Ambiente, Alimenti (AAA), Università del Molise, via de Sanctis snc, 86100 Campobasso (CB)

In vitro model system of micropropagated microshoots of the commercial clone ‘Villafranca’ of Populus

alba (L.) was used to explore the metal phytoremediation potential of poplar. The aim of this work was to

investigate accumulation, translocation and tolerance of autotrophic clones Populus alba in response to high concentrations of cadmium (Cd). For this purpose, increasing concentrations of Cd (0, 5, 50 and 250 µM) were administered to micropropagated poplar plantlets exposed to metal treatments for 15 days. To evaluate Cd influence on Poplar after Cd exposure, was revenue the following morphometric and chemical parameters: number of leaves, length of roots and stems, leaf area index (LAI), shoot and root dry biomass, shoot and root Cd content as well as cadmium influence on essential nutrients content in the vegetable tissues. At the end of the treatment, the seedlings were sampled, weighed and finally mineralized after drying in a stove for the determination of Cd content in the respective underground and aboveground portions. Results obtained in this in vitro screening showed various potential of investigated clone for cadmium phytoremediation. Stability of investigated parameters was most expressed by ‘Villafranca’ clone at low and moderate amounts of metal (5µM and 50µM CdSO4) with which Poplar

reacted with highest growth and lowered accumulation of Cd. Seedlings exposed to the highest pollution concentration (250 µM) show more severe effects. Results show that cadmium has affected the development of the seedlings especially at the highest concentrations. The contaminant has influenced drier biomass plants although the plants generally showed good tolerance to metal treatments. However, the tolerance indexes (Ti) were usually up the value 1, suggesting a good tolerance mechanism of poplar under Cd-exposure. Whereas, the plants had higher translocation factor (TF). Cadmium also seemed to affect absorption of some essential nutrients such as Ca, K, Mg, Mn, S, Fe, Ca, Na, Zn and P, limiting and/or changing their absorption. According to these results, it was shown Populus alba L. had strong tolerance and accumulation capability to Cd, therefore it is a potential Cd-hyperaccumulator.