Multivariate Analysis of Sites Using Water Invertebrates and Land use as Indicators of the Quality...

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Environmental Bioindicators, 2:161–171, 2007Copyright © Taylor & Francis Group, LLCISSN: 1555-5275 print/ 1555-5267 onlineDOI: 10.1080/15555270701590974

UEBI1555-52751555-5267Environmental Bioindicators, Vol. 2, No. 3, Aug 2007: pp. 0–0Environmental Bioindicators

Multivariate Analysis of Sites Using Water Invertebrates and Land use as Indicators of the

Quality of Biotopes of Mediterranean Relic Plant (Petagnaea gussonei, Apiaceae)

Indicators of biotopes quality of Petagnaea gussoneiDe Castro et al. OLGA DE CASTRO,1 LORENZO GIANGUZZI,2 PAOLO COLOMBO,2 PAOLO DE LUCA,1 GIUSEPPE MARINO,1 AND MARCO GUIDA1

1Università degli Studi di Napoli Federico II, Napoli, Italy2Università degli Studi di Palermo, Palermo, Italy

Petagnaea gussonei is a rhizomatous, perennial scapose hemicryptophyte speciesbelonging to a monospecific genus of Apiaceae. It lives only along woodland rivuletsand is a Tertiary relic localized in the restricted area of the Nebrodi Mountains (NESicily, Italy). Ecological indicators have been employed to understand the status ofeight localities (intermittent rivulets) where the species occurs. In order to evaluateboth the rivulets’ water-quality and landscape use, index of anthropisation (Ia), waterfleas bioassay (TU), and analyses of water macroinvertebrates have been employed.The results obtained for the level of toxicity exclude the presence of macropollutants(TU < 0.001), whereas the analyses of the macroinvertebrates showed the occurrenceof several microorganisms typical of altered environments. Correlations of the resultsabout macroinvertebrates with those related to index of anthropisation for each siteare positive. Levels of Ia range from 1.29 (minimum anthropisation) to 3.01 (maximumanthropisation), with a mean of 2.41. On the basis of such results, we can confirm theeffectiveness of using macroinvertebrates as bioindicators to analyze the status ofwater together with the other indices used to examine the ecological conditions of thebiotopes where the plant occurs.

Keywords: anthropisation index, Daphnia bioassay, intermittent rivulets, macroin-vertebrates analysis, Petagnaea gussonei, Sicily

Introduction

Petagnaea gussonei (Sprengel) Rauschert (Petagnia saniculifolia Guss.) is a perennialsmall plant (Figure 1) occurring in 15 small and vulnerable relic Tertiary populations in arestricted area of Italy (Nebrodi Mountains, NE Sicily) (Figure 1) (Wolff 1913). The rhi-zomatous species belongs to a monospecific genus of Apiaceae, is a scapose hemicrypto-phyte and lives along woodland rivulets. Its extremely scattered distribution in a ratherscanty area seems to be related to its stenoecy (adaptation to restricted environmental con-ditions). In fact, the entity grows in shade stations of small mountain rivulets, on hydro-morph lands which allows for considerable water retention, a fundamental requisite for the

Address correspondence to Olga De Castro, Università degli Studi di Napoli Frederico II, Dip.delle Scienze Biologiche, Sez. Biologia Vegetale, Via Foria 223, I–80139 Napoli, Italy.E-mail: [email protected]

162 De Castro et al.

survival of this species (Figure 2) (Brullo et al. 1976, Gianguzzi et al. 2004). P. gussoneiusually reproduces asexually through stolons, horizontal branches growing out from thebase of the plant that produce new plants from buds at their tips (Gianguzzi and DeCastro, unpublished data). Such a reproductive mechanism allows the plant to colonizethe wet edges of rivulets. Seed production occurs occasionally and the germination rate isvery low.

P. gussonei has been classified as EN (Endangered) according to the InternationalUnion for Conservation of the Nature and Natural Resources (IUCN) Red List Criteria,because there are only a few populations scattered over an area of approximately 400 km2.In addition, it is believed that increased water use in the area for human purposes drawswater away from the species habitat, resulting in a natural decline of the area, habitat qual-ity, number of populations and individuals. The species is listed in Appendix I of the BernConvention and in Annexes II and IV of the EC Habitats Directive.

The goal of this study was to evaluate methodologies which may be used to character-ize the habitat where P. gussonei grows and provide information on criteria to adopt inorder to protect this endangered taxon.

The assessment of plant landscape use was performed with the index of anthropisa-tion (Ia) as employed in Mancini et al. (2005). Daphnia magna Straus (water fleas) immo-bilization test was used as a low-cost preliminary screening method to detect the presence

Figure 1. Geographical distribution of Petagnaea gussonei in Italian Peninsula (Sicily). The areais marked with � and is localized in the Nebrodi Mountains. The area corresponds to about 400km2 in which eight intermittent small rivulets have been analyzed where P. gussonei occurs.Water and macroinvertebrates for each rivulet have been sampled. Present information about thesites in the study can be found in Table 1. The graphical description of P. gussonei is presentwithin Sicily.

Indicators of Biotopes Quality of Petagnaea gussonei 163

of macropollutants in the spring waters (Dodson and Hanazato 1995, Kikuchi et al. 2000).In addition, the macroinvertebrates were sampled and examined because they can be usedas bioindicators (Brown et al. 1997).

Materials and Methods

Study System

The studied area is about 400 km2 in the Nebrodi Mountains (NE Sicily, Italy) (Figure 1).Nebrodi Mountains are characterized by the asymmetry of each side, different modellingof the landscape, very rich vegetation and humid environments (Gianguzzi, 1999). In thisarea eight intermittent rivulets have been sampled (Table 1). The survey of the localitiesunder study was carried out in the Autumn 2005. Several field investigations were per-formed from Spring 2004 to Autumn 2005 to examine the amount of water and the micro-fauna and to verify the accessibility to rivulets. Ecological, phytosociological andgeoserial features of sites studied are widely described in Gianguzzi et al. (2004), with theexception of two localities, only recently individuated (sites 2 and 6 in Table 1).

Figure 2. Scheme of the vegetation along Vallone Calagna (site 7) where Petagnaea gussoneioccurs (from Gianguzzi, 2002): 1) P. gussonei community (Petagnaeetum gussonei); 2) hygrophy-lous woodland of Populus nigra and Salix pedicellata (Ulmo canaescentis-Salicetum gussonei); 3)hazel grooves; 4) deciduous oak woodlands of thermophylous type (Oleo-Quercetum virgilianae).

164

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Toxicity Test

The organism used for water toxicity test was Daphnia magna, a Crustacean (Cladocera)extensively used for tests related to aquatic toxicology for its small size, short life cycleand amenability to lab culture. D. magna is the most sensitive in test aiming to measurepollutants and is often used as a test organism for environmental risk assessment. The testwas performed according to the International Organization for Standardization (ISO 6341,1989). Young females (age 0–24 h) were incubated in freshwater samples, in the dark. Thenumber of immobile water fleas was recorded after 24 h to determine the concentrationneeded to achieve 50% immobilization (EC50). Raw data were analyzed using the soft-ware ToxCalc® version 5.0. Toxicity data were converted to Toxic Units (TU) using theformula: TU=100/L(E)C50.

All the bioassays were performed under static conditions, with no renewal of the testsolution. In order to check the correct procedure and sensitivity of the test, a reference testfrom time to time is required. Quality control tests were performed with the referencetoxicant potassium dichromate (Sigma-Aldrich), using the following dilution series:3.2-1.8-1-0.56-0.32 mg/l.

Assessment of Biological Communities

The sampling of the macroinvertebrates was difficult because most of the rivulets werevery narrow (about 15–80 cm wide). For these reasons each rivulet was sampled in moresites. The macroinventerbrates were collected with a handnet exploring bottom substratasuch as stones, mud and macrovegetation. All collected material was placed in plastic con-tainers and fixed with ethanol (70%). In the laboratory, animal classification was per-formed using a stereoscopic microscope (Wild Heerbrugg 374590) using identificationkeys (Sansoni 1988, Campaioli et al. 1994). Identified macroinvertebrates were comparedwith literature data (Campaioli et al. 1994). According to Campaioli et al. (1994), thegroup of Plecoptera is correlated with a very good quality index of water, the macroinver-tebrates Ephemeroptera and Trichoptera are correlated with good quality index of water,and other groups, such as Irudinea, Anelida, and Diptera, are representative of bad qualityindex of water.

Plant Landscape Use

The land use data were established with anthropisation index (Ia) obtained fromCORINE Land Cover database (Krynitz 2000). CORINE category for land use wasretrieved for 1 km radius area, around each sampling site, using the cartographies ofvegetation (scale 1:10000), some of which published in Gianguzzi et al. (2004), othersexpressly carried out (Gianguzzi, unpublished data). The index was calculated as Ia =∑kipi where ki is the specific coefficient for each CORINE land use category and pi isthe relative frequency of each category. The k values were attributed to different catego-ries: 1 - natural woods (with Fagus sylvatica L., Quercus cerris L., Quercus virgiliana(Ten.) Ten., Ostrya carpinifolia Scop.); 2 - meadows and shrubbery; 3 - agriculturalareas (hazel, olive, and orange groves); 4 - urban areas (including road system). Theindex ranged from 1 (minimum change in land use) to 4 (maximum change in land useto human-dominated systems).


Results

Toxicity Test

Daphnia bioassay shows that analyzed rivulets do not present macropollutants as pesti-cides, heavy metals and so on. Toxicological values for each site are not significant, beingTU < 0.001 (Table 2).

Assessment of Biological Communities

According to macroinvertebrates collections, a total of 14 taxa (52 macroinvertebrates)were collected from the eight sites in study (Table 3). Macroinvertebrates distribution isnot constant in the several sampling sites. The community varies from one taxon inTortorici (site 1) to four taxa in other sites, as shown in Table 3. The rivulets are domi-nated by Artropoda, with Diptera primarily, and afterwards by Ephemeroptera, Plecopteraand Trichoptera; other invertebrates groups (Anelida and Gastropoda) were present butnot abundant. Briefly, we summarized the results for each site in Appendix 1.

Plant Landscape Use

Values for Ia ranged from 1.29 (site 3) to 3.01 (site 1), with a mean value of 2.41 (Table 2and Figure 3). Only sites 3 (Torrente Contrada Contrasto), 5 (Torrente Galati-San Basilio),and 8 (Vallone Case Mangalaviti) had predominantly natural conditions for growth ofP. gussonei. In particular, in the first and in the last site, the natural woodland aspects pre-dominate, whereas in the second one shrublands and meadows are present for the mostpart. The other sites are localized in prevalently agricultural areas (especially hazel andchestnut groves), with values of incidence near 80% in the sites 4 (Torrente Fiumetto) and6 (Torrente Monte Cuculone) or even higher in the sites 1 (Fiumara Tortorici), 2(Castell’Umberto), and 7 (Vallone Calagna). On the whole, in the last one, urban areas

Table 2Sampling sites with Italian name and results using the toxicity test with Daphnia biossay

Code Site Water-pH Toxic UnitsIndex of

anthropisationBioindicatorswater-quality

1 Fiumara Tortorici 6.87 < 0.001 3.01 +2 Castell’Umberto 7.13 < 0.001 3 −3 Torrente Contrada

Contrasto7.19 < 0.001 1.29 +

4 Torrente Fiumetto 7.67 < 0.001 2.71 −5 Torrente

Galati-S. Basilio7.81 < 0.001 1.98 +

6 Torrente Monte Cuculone

7.21 < 0.001 2.77 −

7 Vallone Calagna 7.74 < 0.001 2.86 −8 Vallone Case

Mangalaviti7.16 < 0.001 1.68 +

Assessment of biological communities uses the macroinvertebrate as bioindicator (±).


have a great influence and are represented especially where the water-course laps thevillage of Tortorici.

Discussion

It was evident from our study that the analysis of ecological indicators may provideimportant information for the understanding of vulnerable areas. In this case, no ecologi-cal study had been carried out on the area where P. gussonei occurs. In addition to normalindices used (Ia and TU), we have tried to analyze only the macroinvertebrates living inthe rivulets, since it is not possible to use Extended Biotic Index (EBI) or the Italian indexof EBI (Woodiwiss, 1981, Ghetti, 1997), because these can not be used for intermittentsmall rivulets. The choice to consider the macroinvertebrates has been justified, becausethese constitute an important component of biodiversity in a lotic system; in addition theyare very sensitive and respond to both natural and man-induced changes in their environ-ment as reported by De Pauw and Hawkes (1993).

Table 3Presence of macroinvertebrates taxa in the eight sites in study

Sites and presence of macroinvertebrates

Taxa 1 2 3 4 5 6 7 8

AnnelidaHirudinea

Limnatis sp. XOligochaeta

Lumbricidae X XArtropodaColeoptera

Elminthidae XHelodidae X

DipteraCeratopogonidae XChironomus sp. X X XDixidae XTabanidae X

EphemeropteraBaetis sp. X X XElectrogena sp. X

PlecopteraCapnia sp. X X X XXantoperla sp. X

TrichopteraHydropsychidaePhilopotamidae X X X

GastropodaAncylus sp. X


In detail, this study has revealed that in the localities where P. gussonei occurs, thereare traces of the antrophic impact, as it is obvious for example near Torrente Calagna (site 7)(Table 2 and Figure 3). The analyses carried out in this area showed an elevated index of

Figure 3. Schematic representation of four COoRdination of INformation on Environment (CORINE)land use categories (X-axis) and their relative frequency (y-axis) for the eight sites in study. Value ofthe index of anthropisation (Ia) is present for each site. Ia=∑kipi, where ki is the specific coefficientfor each CORINE land use category and pi is the relative frequency of each category.


anthropisation (equal to 2.86, with 88% of agricultural areas), against only 15% of naturalwoods (Figure 3). Consequently, we can also see a very low structure of microfauna, sinceonly one kind of organism of the Coleoptera family has been found. However, the com-forting, negative result of the analysis on the level of toxicity through the Daphnia bioas-say excludes the presence of macropollutants. This result is confirmed for all the othersites studied (Table 2).

The results of the analyses of the macroinvertebrates are not very good. The check ofdifferent microrganisms, typical of altered environments and not very organized commu-nities (see Appendix 1). Consequently, if the test of the Daphnia bioassay is negative forthe rivulets in study (i.e., no macropollutant), the obtained results from the study of macroin-vertebrates might be considered a potential sign of danger for the qualitative status of theareas under study. In fact, correlating the results of the examined macroinvertebrateswith the index of anthropization of each site, we can see a positive correlation between thisone and the status of an eventual degradation of the waters, except for the site 1 (FiumaraTortorici, Table 2).

From what has been examined in situ and from the obtained results from our analyses,it turns out that the populations of P. gussonei are critically threatened since the greatestdamages until now are due to the uptakes of the sources and of the stratums (generally forhuman utilizations) as well as to the use of weedkillers for the plantations as also con-firmed in Gianguzzi et al. (2004). Among the other observed alterations, the deforesta-tions, the reforestation with exotic species, the illegal dumps in the watercoursesthemselves, the opening of roads and so on can be mentioned. On this basis together withthe data collected by our analyses, we can reconfirm that the conditions of preservation ofP. gussonei in the Nebrodi Mountains area are rather vulnerable. To reduce the evidentrisks which lie with the species, it would be opportune to submit the different stations to amore careful protection since some of them are not included in protected areas. On thissubject an interesting proposal could be to incorporate these stations in the Nebrodi Park,in fact from the obtained results it turns out clearly that the stations which are included inthe Park are not damaged (Table 1 and Table 2).

In conclusion, we would like to note that this study is not intended to show a prefer-ence for these approaches to others for the study of the habitat, but to show that a wholerange of scientific approaches is fundamental in order to have a good knowledge of aregion where there are endangered specimens as P. gussonei.

References

Brown AV, Aguila Y, Brown KB, Fowler WP. 1997. Responses of benthic macroinvertebrates insmall intermittente streams to silvicultural practices. Hydrobiologia 347:119–125.

Brullo S, Grillo M, Guglielmo A. 1976. Osservazione ecologiche preliminari su Petagnia saniculifo-lia Guss., raro endemismo siculo. Giornale Botanico Italiano 110(4–5):293–296.

Campaioli S, Ghetti PF, Minelli A, Ruffo S. 1994. Manuale per il riconoscimento dei macroinverte-brati delle acque dolci italiane. Trento: Provincia Autonoma di Trento. 357 p.

De Pauw N, Hawkes HA. 1993. Biological monitoring of river water quality. In: Walley WJ, Judd S,editors. River water quality monitoring and control. Birmingham: Aston University, p 87–111.

Dodson SI, Hanazato T. 1995. Commentary on effects of anthropogenic and natural organic chemi-cals on development, swimming behaviour, and reproduction of Daphnia, a key member ofaquatic ecosystems. Environ Health Perspect 103(S4):7–11.

EC Habitats Directive. Council Directive 92/43/EEC. Available from: http://ec.europa.eu/environment/nature/nature_conservation/eu_nature_legislation/habitats_directive/index_en.htm. AccessedMay 2007.


Ghetti PF. 1997. Manuale di applicazione: Indice Biotico Esteso (I.B.E.). I macroinvertebrati nel con-trollo della qualità degli ambienti di acque correnti. Trento: Provincia Autonoma di Trento. 222 p.

Gianguzzi L. 1999. Flora e vegetazione dei Nebrodi–itinerari didattici. Regione siciliana. Messina:Arti Grafiche Zuccarello. 233 p.

Gianguzzi L. 2002. Osservazioni sulla conservazione in situ di Petagnaea gussonei (Spreng.) Rauschert,paleoendemita esclusivi dei Monti Nebrodi (Sicilia nord-orientale). Inform Bot Ital 34:63–69.

Gianguzzi L, La Mantia A, Lo Presti RM. 2004. Distribuzione, ecologia e status conservativo diPetagnaea gussonei (Sprengel) Rauschert (Apiaceae). Naturalista Sicil 19(1)(S4):205–242.

International Organization for Standardization 6341, 1989. Water quality-Determination of the inhi-bition of the mobility of Daphnia magna Straus (Cladocera, Crustacei). ISO 6341, 01.10.1986.

Kikuchi M, Sasaki Y, Wakabayashi M. 2000. Screening of organophosphate insecticide pollution inwater by using Daphnia magna. Ecotoxicol Environ Saf 47:239–245.

Krynitz M. 2000. Land Cover. Annual Topic Update 1999. Copenhagen: European EnviromentAgency. 25 p.

Mancini L, Formichetti P, Anselmo A, Tancioni L, Marchini S, Sorace A. 2005. Biological quality of run-ning waters in protected areas: the influence of size and land use. Biodiversity Conserv 14:351–364.

Sansoni G. 1988. Atlante per il riconoscimento dei macroinvertebrati dei corsi d’acqua italiani.Trento: Servizio protezione ambiente. 190 p.

ToxCalc® 1996. User’s guide. Version 5.0. McKineyville, CA: Tidepool Scientific Software.Woodiwiss FS. 1981. Biological water assessment methods. Nottingham-Abridged report of work-

ing group expert, Commission of the European Communities, ENV/416/80.Wolff H. 1913. Umbelleferae-Saniculoideae. Das Pflanzenreich 4(228):22–26, 273–274.

Appendix. Results of assessment of biological communities for each site instudy

Code Site Description

1 Fiumara Tortorici The station presents a sufficiently structured community for the presence of two different genus of Plecoptera which are typical of well-oxygenized and unpolluted environments. This information may give evidence of an unpolluted or unaltered environment

2 Castell’Umberto It presents a small community characterized by organisms able to colonize habitats rich in organic detritus; the two species found live in slow flowing or stagnant waters. This aspect and the absence of organisms typical of well structured environments imply an unfavorable judgment and mark a polluted and altered environment

3 Torrente ContradaContrasto

The station presents a structured community of several taxa. Besides the presence of individuals of the genus Capnia, that are typical of well oxygenized and unpolluted environments, determines a sufficient judgment

4 Torrente Fiumetto Taxa belonging to the genus Chironomus and Tabanidae’s family have been found in this site. These organisms are typical of habitats with high organic contamination. For these reasons it is possible to assert that it is very polluted or altered

(Continued)


Appendix. (Continued)

Code Site Description

5 Torrente Galati- S. Basilio

It presents a sufficiently structured community with Plecoptera, Ephemeroptera and a relative abundance of individuals belonging to the family of Philopotamidae that are typical of well oxygenized springs. The judgment may be positive and emphasizes an unpolluted or not considerably altered environment

6 Torrente Monte Cuculone

The station presents a community mostly composed of organisms which prefer highly organic contaminated habitats. Usually Ephemeroptera specimens give evidence for good water quality unlike the presence of Baetis sp. which shows a discreet tolerance to pollution. This aspect, together with the presence of typical polluted environmental forms, determines an unfavorable judgment and marks a polluted or altered environment

7 Vallone Calagna The environment appears characterized by microfauna which includes species of Elmintidae’s family, strictly correlated to torrential, well oxygenized habitats. However the station does not have a structurally organized community and it is possible to assert that it is altered

8 Vallone Case Mangalaviti

The station presents a sufficiently structured community, with Plecoptera and Ephemeroptera taxa. These are indicators of a very good quality of superficial current waters, especially for specimens of genus Capnia. In general the judgment may be positive

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