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Towards guidelines for monitoring threatened species of amphibians andreptiles in ItalyGiuseppe M. Carpanetoa; Marco A. Bolognaa; Riccardo Scaleràa Dipartimento di Biologia, Università Roma Tre, Roma, Italy

Online publication date: 22 November 2010

To cite this Article Carpaneto, Giuseppe M. , Bologna, Marco A. and Scalerà, Riccardo(2004) 'Towards guidelines formonitoring threatened species of amphibians and reptiles in Italy', Italian Journal of Zoology, 71: S1, 175 — 183To link to this Article: DOI: 10.1080/11250003.2004.9525555URL: http://dx.doi.org/10.1080/11250003.2004.9525555

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Towards guidelines for monitoring threatened species of amphibians and reptiles in Italy

GIUSEPPE M. CARPANETO MARCO A. BOLOGNA Dipartimento di Biologia, Università Roma Tre, Viale .Marconi 446, 1-00146 Roma (Italy) E-mail: boIogna@bin.uniromaSit, carpanet@uniroma3U

RICCARDO SCALERÀ Via Valentino Mazzola 38, 1-00142 Roma (Italy)

ABSTRACT

After an extensive review of the ecological literature on the Ital­ian amphibians and reptiles, the authors furnish the guidelines for monitoring the most threatened species, discussing criteria and methods suitable to seven major groups defined in taxonomic and ecological terms: salamanders, newts, frogs, terrapins tortoises, lizards and snakes. Each major group is analysed separately, in or­der to choose the most adequate techniques for achieving infor­mation on: (1) habitat distribution and preference, (2) population size and density, (3) home range size, (4) reproductive biology, (5) use of space and time, (6) feeding habits. The aim of the pa­per is to provide wildlife managers with tools and knowledge for conserving amphibians and reptiles at regional and national level.

KEY WORDS: Amphibians - Reptiles - Italy - Monitoring -Wildlife Management - Conservation

ACKNOWLEDGEMENTS

We are grateful to all the authors of the scientific papers pub­lished on this special issue of the Italian Journal of Zoology, ded­icated to the conservation status of threatened amphibian and reptile species of the Italian fauna. In fact, the guidelines given in this paper have been deduced from their contributions to single species. Moreover, we thank all the members of the Italian Her-petological Association (Societas Herpetologica Italica) who im­proved this work through a direct or indirect exchange of infor­mation and ideas. A special thank is given to the Direction of Na­ture Protection (Ministero dell'Ambiente e della Tutela del Territo­rio) who promoted and financed the monitoring research.

INTRODUCTION

The present work was realized for the project "Moni­toring the conservat ion status of the most threatened amphibian and reptile species of the Italian fauna", car­ried out by the Department of Biology, University "Ro­ma Tre", and promoted by the Direction for Nature Pro­tection, Ministry of Environment (convention on 29 No­vember 1999). The aim of this paper is to provide an in­t roduct ion to the guidel ines for moni tor ing the most threatened species of amphibians and reptiles, in accor­dance with the provisions of the DPR no. 357, 8 Sep­tember 1997 (Anicle 7), which regulates the application of the Directive 92/43/EEC.

Amphib ians and reptiles are important e lements of our national biological heritage and represent significant segments of the evolutionary history of the Italian fau­na. They are crucial to the natural functioning of many ecological p rocesses and key componen t s of the ecosystems, either as preys or predators. For all these reasons, amphibians and reptiles deserve special atten­tion in wildlife management. An updated knowledge of their populat ion status and ecology represents a basic step for any intervention on wildlife.

Baseline information of the status and health of Italian popula t ions of amphibians and reptiles is remarkably sparse. Only some regions or provinces carried out atlas projects focused to gather information on past and pre­sent distribution of these animals at small scale. During the last decades, many Italian populat ions of amphib­ians and reptiles showed a remarkable decline. In fact, many researchers and local observers suggested a de­creasing of population density or a local extinction for several species in the areas they are usual to monitor. Really, some species disappeared from many Italian lo­calities w h e r e they occurred in the past decades , e.g. the fire salamander {Salamandra salamandra) and the yellow-bellied toad {Bombina variegata) from several areas of central Italy (Bologna et al., 2000). These cases of local extinction are probably due to a combination of natural and human-caused factors that worked together producing declines from which some populations could not recover and thus they went to extinction. There is a general agreement among scientists that amphibian and reptile populations are declining mainly because of hu­man-induced factors such as the loss of habitats (espe­cially wet zones and small ponds), the transformation of grasslands into cultivated fields, the pollution of water and the spread of pesticides which reduced their poten­tial preys, i.e. the invertebrate communities, and the in­troduction of non-native predators, such as alien fishes and freshwater turtles. All these factors are due to hu­man activities despite of the law provisions to protect wildlife a national and regional level (Gentilli, 2004; Scalerà, 2004; Scoccianti, 2004). Nevertheless , t he above-ment ioned human-caused factors may intensify natural factors, such as global climatic changes and the consequent lowering of the water-bearing stratum. Ac-

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176 G. M. CARPANETO, M. A. BOLOGNA, R. SCALERÀ

cording to many researchers, although the effect of global climate change on many species, especially am­phibians, is speculative, it has the potential for causing the loss of many species. Drought and the increased ul­traviolet radiation reaching the ground seem to be the major factors adversely affecting amphibian populations. However, it should be emphasized that amphibian and reptile populations also may vary in size because of nat­ural factors, particularly extremes in the weather. The size of their populations may vary, sometimes dramati­cally, from year to year, so what is perceived, as a de­cline may be pan of long-term fluctuations.

A profound need exists for national coordination of regional inventories and population studies, including a national effort to monitor amphibians on parks, national forests, wildlife refuges, and other public lands. Only through long-term monitoring studies will better data on population changes through time and between sites become available. Such data are essential to evaluating the population status and trends of amphibians and rep­tiles in Italy, but also for determining the impact of nat­ural and human-caused factors on the different life-his­tory stages of these animals. Likewise, understanding the roles which amphibians and reptiles play in aquatic and terrestrial ecosystems is essential. It is obvious that early detection of problems is crucial to successful re­medial action. In many ways, a national program of monitoring amphibian and reptile populations is like preventive medicine; the earlier a problem is detected, the greater the likelihood of successful treatment and the lower the cost. A proactive national program based on standardized scientific methodology and applied across all species and habitats will go a long way to­ward ensuring that amphibians and reptiles remain a healthy component of our national biological heritage. Detailed work on the ecology of species and the factors implicated in decline needs to be a continuous work in order to plan a wildlife management based on adequate scientific data and careful planning.

Monitoring research has the following objectives: (1) to define the population status, the detailed distribution and the ecological role of the species in the study area; (2) to record the habitat changes caused by human activities and to suggest their eventual effects on animal popula­tions; (3) to assess die population trends of rare or declin­ing species; (4) to assess the impact of eventual manage­ment decisions on the species and their habitats, as well as the economic costs and human resources required.

To define the population status, researchers should gather data on population size, density and dynamics, sex ratio and age structure, but also on the ecological role of the species in the community (details on home range size, foraging activity, preferred food items, preys and predators).

Methods for monitoring amphibians and reptiles de­pend on the life style of the species. The most adequate methods have to be chosen for each study case, in ac­cordance to the habits of the species, its habitat type and

life cycle. Other factors influencing the method choice are financial budget, time and human resources at dis­posal. Survey recurrence (daily, weekly, monthly, etc.) and period (one season or more) should be defined by the expert charged of the monitoring programme, and may vary in accordance to the phenological features of the species in the study area. Nevertheless, all the suc­cessive monitoring activites should be conducted using the same method, in order to compare results and obtain a long-term scenario of population dynamics. The same methods, as much possible, should be used for monitor­ing a single species in different regions of the country, in order to obtain patterns and trends of population ecolo­gy for that species at national level.

Recruitment of experts is a very important aspect to consider. Personnel must be chosen among specialists having a documented experience on population ecolo­gy of the target species or of other species belonging to the same taxonomic group. Owing to the difficult iden­tification of some species during fieldwork, especially juvenile specimens or larval stages, the expert should be an experienced herpetologist other than a popula­tion ecologist. Therefore, the management authorities should turn to the Italian Herpetological Association, in order to obtain information on the specialists.

The life habits of the amphibians are very different ac­cording to the species because of their ecological spe­cialization. The adults of some species of Urodela have an active life that takes place exclusively or mainly in water {Proteus, Triturus, Euproctus); other species live mainly or exclusively in terrestrial habitats (Salamandra, Salamandrina, Speleomantes). Also among the anurans (frogs and toads) there are some remarkable differences between the more aquatic (e.g. Rana, Discoglossus, Bombino) and the more terrestrial species (e.g. Bufo, Hyld). These differences exert an influence on the choice of the methods for monitoring the populations.

In the present paper, monitoring criteria and techniques were discussed separately after the definition of seven groups based on a combination of taxonomical and eco­logical features: (1) terrestrial salamanders; (2) aquatic salamanders (newts); (3) frogs (including all the anurans); (4) tortoises (terrestrial turtles), (5) terrapins (aquatic tur­tles); (6) lizards; (7) snakes. Sea turtles were excluded from this analysis because they belong to marine ecosys­tems and should be studied in a very different ambit, i.e. conservation and exploitation of marine resource.

An important synthesis of the studies for monitoring amphibians was offered by Heyer et al. (1994). Other relevant works have been published on the techniques for studying amphibians and reptiles in the field. Some of them are listed herein and grouped according to their specific subject: (1) General methodology (Camp­bell & Christman, 1982; Scott, 1982; Bury & Raphael, 1983; Peterson & Dorcas, 1992; Pianka, 1993; Vitt & Pi-anka, 1994); (2) toe clipping (Hero, 1989; Waichman, 1992); (3) pitfalls e drift fences (Gibbons & Semlitsch, 1982; Bury & Corn, 1987; Mengak & Guynn, 1987); (4)

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GUIDELINES FOR MONITORING THREATENED SPECIES 177

microchip marking (Camper & Dixon, 1988); (5) mark­ing (Kaplan, 1958; Clark, 1971; Emlen, 1988); (6) stom­ach flushing and other techniques for studying the diet (Legler, 1977; Legler & Sullivan, 1979; James, 1990; Ledere & Courtois, 1993; Carretero & Llorente, 2001).

In the last decades, many Italian zoologists have worked on faunistics and ecology of amphibians and reptiles, determining a great improvement of our knowledge. A specific database concerning amphibians and reptiles at national level was realized by the Italian Herpetological Association, who has published the pre­liminary results and distribution maps (Societas Her-petologica Italica, 1996); the final volume of the Atlas Project will be published in the next year. These records were also utilized by the Department of Nature Protection of the Ministry of Environment, to construct a national faunistic database. In the mean time, several Atlas Projects at local level (regions, provinces or mu­nicipalities) have been realized.

Moreover, three important publications dealing with the Italian fauna appeared in the last decade, under the promotion of the Ministry of Environment. The first one (Minelli et ai, 1993) is a Checklist of the species of the Italian fauna, a true milestone for the future zooological research; the second one (Minelli et ai, 2000) is a com­prehensive overview of the Italian fauna and habitat types; the third one (D'Antoni et ai, 2003) provides in­formation and distribution maps for all the animal species included in the Habitat Directive (1992). Anoth­er important publication was promoted by the Ministry of Agriculture and Forestry Politics (Scalerà, 2003) and is a generic treatise on the threatened species of Italian amphibians and reptiles.

Obviously, the data for all such summaries come from the activities of many Italian researchers working with various aspects of the biology of amphibians and rep­tiles and are found in many scientific publications. To il­lustrate these research efforts by examples, we quote the following papers that provide information on sever­al ecological aspects of these animals, and represent a collection of useful data for their monitoring and man­agement: Giacoma, 1983; Cheylan, 1984; Andreone, 1986; Barboult & Mou, 1988; Pavignano, 1988, 1990; Av-ery & Perkins, 1989; Andreone et al., 1990, 1994, 1999; Pavignano et al., 1990; Luiselli & Agrimi, 1991; Capula et al., 1992, 1993, 1994, 2002a, 2002b; Fasola & Canova, 1992; Joly & Giacoma, 1992; Massot et al., 1992; Capula & Luiselli, 1993, 1994a, 1994b, 2002; Giacoma et al., 1993; Rugiero, 1994; Luiselli et al., 1994, 1996, 1997; Gentilli & Zuffi, 1995; Angelici et ai, 1997; Salvidio, 1998; Bressi et al., 1999; Capizzi, 1999; Capula, 1999; Luiselli & Capizzi, 1999; Mazzoni, 1999; Salvidio et al., 1999; Van Damme, 1999; Filippi & Luiselli, 2000; Guidali et al., 2000; Hailey, 2000; Corti et al., 2001; Zuffi, 2001; Bombi & Bologna, 2002; Mazzotti et ai, 2002.

The present paper was written after a critical review of many scientific papers devoted to ecology of am­phibians and reptiles. In particular, the guidelines pro­vided herein were deduced from the contributions giv­

en to the knowledge of single species by the following authors, and published on this special issue of Italian Journal fo Zoology: Andreone et al. (2004a): Salaman­dra lanzai; Scillitani et ai (2004a): Triturus italicus; Bressi (2004a): Proteus anguinus; Andreone et ai (2004b): Pelobates fuscus; Salvidio et ai (2004a): Pelodytes punctatus; Barbieri et ai (2004): Bombino variegata-, Barbieri & Bernini (2004): Rana latastei; Mazzotti (2004): Testudo hermanni; Zuffi (2004): Emys orbicularis; Scillitani et al. (2004b): Cyrtopodion kotschyi; Bressi (2004b): Algyroides nigropunctatus; Di Già & Sindaco (2004), Lapini & Dall'Asta (2004): Lacerto agilis; Salvidio et al. (2004b): Timon lepidus; Lapini et al. (2004): Lacerta horvathi; Bombi & Vignoli (2004): Archaeolacerta bedriagae. Scalerà et al. (2004): Podar-cis filfolensis; Capula (2004): Podarcis raffonei; Luiselli (2004): Vipera ursinii.

TERRESTRIAL SALAMANDERS

Adults can be detected at sight, when they are walking on the ground, weekly surveys along transects in suitable habitats such as forests near streams or lakes, provide an indication of adult numbers and permit the localization of their foraging areas. The survey period, according to the species, may vary from early spring to early autumn, during rainy days, at evening or nightime. The detection of adults is the sole method of censusing the viviparous species, such as Salamandra atra, S. lanzai and Speleo-mantes spp., that lack a larval cycle. On the contrary, the occurrence of eggs and/or larval forms on the bottom of streams, among detritus, dead leaves, stones or aquatic vegetation, allows to localize reproductive sites. The re­productive period can vary from early spring to summer months, according to the altitude above sea level.

To estimate populations of salamanders, adult speci­mens may be marked using toe-clipping or micro-trasponders. The latter are usually injected into the peri­toneal cavity of the animal, through a special tool. Some of these studies have been conducted on small amphibians and indicate that such applications do not influence the survival rate of the animals. Microtraspon-ders allow to gather many data on amphibian ecology because they permit to follow the life history of many single individuals. Marking specimens leads the workers to obtain the home range size of each individual and the population density of the species. The latter is easi­ly calculated by the ratio between the number of indi­viduals and the surface of the habitats suitable for the species in the study area. Through radio-tracking, one may calculate the dispersion ability of the species and thus verify the connectivity or the isolation of single demes in fragmented habitats.

Dietary studies are possible using a technique named "stomach flushing". The latter is used both for salaman­ders and frogs and consists of the introduction of water into the stomach of the animal by a thin catheter, in or-

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178 G. M. CARPANETO. M. A. BOLOGNA, R. SCALERÀ

der to flush out the food remains. Such method must consider the trophic activity of the animals according to their behavioural ecology and weather conditions. Gen­erally speaking, captures should be made some hours after they started the foraging activity, allowing the ani­mals to catch a certain number of preys. On the other hand, captures should not be made too late, when the animals could have ceased their foraging activity and be digesting. The workers have to make some preparatory observations in order to assess well the foraging activity of the animals as this may vary according to local con­ditions and weather. In order to reduce stress and risk of manipulation, stomach flushing should be executed under anaesthesia (the worker should learn by experi­ence with larger animals before treating the smaller ones). Moreover, the operation should be executed as soon as possible, in order to avoid the deterioration of food remains by digestive enzymes.

An alternative method for censusing salamanders (but also frogs) is the analysis of the colour pattern. In fact, several species of amphibians have a unique pattern of colour, such as the shape of dorsal or ventral spots. The method consists of taking pictures of each individual, on dorsal and/or ventral view, then comparing them with the aid of a computer programme. This kind of analysis fits well to the study of some salamanders e.g. Salamandra salamandra and Salamadrina terdigitata.

AQUATIC SALAMANDERS

Both adults and larvae of Italian newt species should be detected and/or captured in small waterbasins such as ponds, marshes, swamps and channels. Adults may be censused only during the reproductive period when they occur in water. The latter period is varying accord­ing to latitude, altitude, climatic conditions and food availability. The workers have to make some preparatory observations in the study area, in order to establish the reproductive period of a population and to calculate the duration of its permanence in water. In accordance to the latter and owing to newt concentration in small wa­terbasins, the sample period can be reduced to a low number of surveys. The animals can be captured by fish­net or by special traps located on the water surface. These are simple segments of plastic tubes floating on the water surface, where newts can enter but not escape.

There are three methods for estimating population size and density of newts, according to habitat typolo­gy: (1) a complete census can also be made in small basins with transparent water, where all the individuals can easily be counted without capturing them; (2) a simple count of the individuals can be made in a small basin with deep or dark water where a team of opera­tors counts all the newts as they rise at the surface of water for breathing, before the second reascent of the first individual observed; (3) a temporary removal of all the individuals can be made in small biotopes such as

an isolated pond or a drinking trough; they can be counted, measured, sexed, marked and then released; (4) marking and recapturing a certain number of indi­viduals is the sole method adopted for large, deep or dark waterbasins, such as marshes, swamps and rivers. The first two methods only give the population size and density; only the third and fourth method allow the worker to gather data on reproductive biology, sex ra­tio, age structure, occurrence of pedomorphic speci­mens and other parameters.

Adult specimens may be marked using toe-clipping or microtrasponders, like the terrestrial salamanders. Micro-trasponders also allow to detect the microhabitats where newts take refuge at the end of the aquatic phase of their annual biological cycle. Population density of the species may be calculated as the ratio between the num­ber of individuals and the surface of the waterbasin or the estimated water volume. Through toe-clipping or ra­dio-tracking, one may calculate the dispersion ability of the species on the ground and thus verify the connectiv­ity of single demes occupying isolated ponds. All the Italian newt species fit well to censusing techniques based on the analysis of the colour pattern. In fact, all the Italian species have a spotted belly that represents a sort of "identity card" for each individual. Dietary studies are possible using stomach flushing techniques. Such method should be applied during both day and night-ime, in order to assess the foraging activity of the newts in the study area. Otherwise from terrestrial salaman­ders, the feeding activity of newts is less influenced by weather conditions, and not closed to day or nightime.

A special case is given by Proteus anguinus, a pedo­morphic species adapted to live in subterranean water­basins, restricted to the most northeastern provinces of Italy (Trieste and Gorizia). The only biotopes where this species can be observed are caves crossed by karstic rivers. Owing to the environmental stability of these subterranean habitats, P. anguinus is active and repro­duces all over the year. Monitoring this species is ex­tremely difficult and it requires a good speleological ex­perience. Only adults are usually captured (juveniles are rarely seen), and sexing the specimens is almost im­possible. No marking method has been tested: toe-clip­ping is not valid because this pedomorphic species shows a rapid regeneration.

ANURANS

The adults of anurans show a diverse degree of water dependence: some species spend most of their time in or around water, like green frogs (Rana spp.), Italian stream frog (Rana italica), common frog (Rana tempo-raria), painted frogs (Discoglossus spp.) and yellow-bel­lied toads (Bombina variegata); other species, on the contrary, go to water only for breeding, like tree frogs (Hyla spp.), toads (Bufo spp.) and the agile frog (Rana dalmatind). The monitoring techniques for the most

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aquatic species are not very different from those de­scribed for aquatic salamanders. They are detected and/or captured in small waterbasins such as ponds, marshes, swamps and channels, but also along the shores of lakes, rivers and estuaries. On the contrary, the most terrestrial species can be censused only during the reproductive period when they rush to water and utter specific calls. The latter period is varying according to species and altitude. The workers have to make some preparatory observations in the study area, in order to establish the reproductive period of a population and to calculate the duration of its permanence in water. As for newts, owing to anuran concentration in small water-basins during the breeding season, the sample period can be reduced to a low number of surveys for the ma­jority of species. An exception to the rule is represented by the yellow-bellied toad (Bombina variegata), which breeds several times from April to August. Breeding of this species usually occurs after some days of pelting rain when a number of temporary ponds are formed. Owing to this intermittent reproduction, tadpoles at var­ious stages of developments, juveniles and adults occur together in the same ponds, from spring to late summer. Therefore, for monitoring this species, frequent surveys should be carried out all over its active life period.

The different methods we described for estimating population size and density of newts, are also valid for anurans, according to habitat typology: the small biotopes may be investigated through a complete cen­sus, in both clear and dark waters, including a tempo­rary removal of all the individuals occurring there. On the contrary, larger and deep biotopes need a mark­ing/recapturing technique or the use of transponders. Toe-clipping is often used also for anurans, and some species fit well to the colour pattern analysis, e.g. the yellow-bellied toad (Bombina variegata). Micro-transponders also provide information on the home range size and foraging activity of terrestrial species like toads, tree frogs and agile frogs. Moreover, they may al­low to detect the microhabitats where the adults take refuge from cold weather (hibernation) or from summer drought (aestivation). For the most aquatic species, pop­ulation density may be calculated as the ratio between the number of individuals and the surface of the water-basin. For the most terrestrial species, on the contrary, the ratio is between the number of individuals and the surface of the habitats suitable for adult activity. Through toe-clipping or radio-tracking, one may calcu­late the dispersion ability of the species on the ground and thus verify the connectivity of single demes occupy­ing isolated ponds or fragmented forests. Dietary studies are possible using stomach flushing techniques. Such method should be applied during both day and night-ime, in order to assess the foraging activity of the anu­rans in the study area. As a rule, frog and toad activities prevail at nightime; nevertheless, many species are also active during daytime, according to weather conditions. Rainy and cloudy days coincide with an increasing activ­

ity of anurans, from springtime to autumn. Surveys for monitoring the spadefoot (Pelobates fuscus) and the Lataste's frog (Rana latastei) in northern Italy are often carried out during the rainy and wet nights, from March to May. The permanence of these two species in their reproductive sites is very short, and surveys must be car­ried out intensively, all together in the same period.

Population estimate of terrestrial species can be made with the aid of drift fences and pitfall traps. This method consists of positioning a plastic barrier (height 50-60 cm) on the ground, namely a drift fence, which causes the animals to drift into pitfall traps. The basic principle behind this trapping method is that when the animals encounter the barrier, they take the route of least resistance by moving either right or left and fol­lowing the fence, up to drop into a pitfall trap, that is a plastic bucket sunk in the ground every 10 m. Traps must be checked every morning at dawn, in order to avoid exposure to predators or dehydration.

Egg and tadpole counts are other methods used for estimating population size. A worker with great experi­ence is able to deduce the numbers of females, which lay in a pond or identify tadpoles without injuring them. The occurrence of tadpoles at advanced development stages is the only evidence for recognizing a reproduc­tive site. Pelodytes punctatus, the most threatened frog of the Italian fauna, is also a species difficult to detect owing to its elusive behaviour. Its calls are very faint and cannot be heard at distance. Although this species breeds twice a year (early spring and autumn), the per­manence of the adults at their reproductive sites may last only few days and is strongly influenced by the sea­sonal rains. For these reasons, the detection of eggs and tadpoles is the only method for monitoring this species. Each female can lay up to 3 egg strings; in any case, the experts recommend to count a female for each group of strings at a distance of 40 cm.

TORTOISES

The Hermann's tortoise is the only terrestrial turtle na­tive of Italy. It is a Mediterranean species whose former range extended along all the littoral and sublittoral zone of Italy. Several human-caused factors as habitat loss, fires, and harvest for commercial purpose, have re­duced the number and density of its populations. The current populations of this species stretch along the Tyrrhenic coast within fragmented areas of holm oak forests, evergreen maquis and gariga. Other populations occur in Sardinia and Sicily, a few also along the Adriat­ic coast. The status of these populations is often uncer­tain because often altered by a number of specimens released from captivity and belonging to alien sub­species (Testudo bermanni boettgeri) or species (7". graeca, T. ibera and others). Several cases of of hy­bridization were observed between Hermann's tortoise and alien species; the latter were also the vehicule for

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180 G. M. CARPANETO, M. A. BOLOGNA, R. SCALERÀ

the introduction of diseases such as some Herpesvirus, which may compromise the health of native popula­tions. The activity period of the Hermann's tortoise is from March to October: in summer, peaks of activities were recorded at dawn and late afternoon, while in springtime these occur in the middle of the day. Repro­duction may occur once or twice every year, at spring­time or late summer.

To check the occurrence of the species and/or find new sites, surveys should be made in the periods of major activity, especially in spring after hibernation (March-June). Individuals are localized at sight, paying attention to the noise they make knocking their carapax during sexual or territorial interactions.

To estimate population size and density, tortoises should be marked and released in the same spot. At least four operators will cross the area walking in paral­lel at a distance of some meters. The best marking method is to make small incisions on the peripherical plates of the scute, according to a conventional code.

TERRAPINS

The only freshwater turtle of the Italian fauna is the European pond terrapin {Emys orbicularis), formerly widespread all over the country but now very local or rare owing to the destruction or alteration of wet zones. In fact, the habitat loss of this species was dramatically reduced by land reclamation for agriculture. The de­struction of small ponds and the deep alteration of marshes and swamps were the man-caused factors in­fluencing the distribution and density of the species.

The annual cycle of the European pond terrapin is strongly influenced by weather conditions: the individu­als may break hibernation in the middle of winter, if weather is favourable and some hot days occur. Popula­tion size and density may be calculated in springtime (March-June) where the species shows the highest peak of activity. During summer, the daily activity becomes bimodal, and the turtles make a break in the middle of the day, during the hottest hours. In summer, they be­come active also at night. Surveys are difficult because of the extreme mistrust of this species which plunges it­self immediately when approached. The workers have to make some preparatory observations in the study area, in order to detect the basking sites and foraging areas, to facilitating both observations and captures. In some areas, owing to turtle concentration in small wa-terbasins, the sample period can be reduced to a low number of surveys. Animals may be counted during basking but can be difficult because of vegetation den­sity. Juveniles are hardly detected because they conceal themselves in the midst of floating and riparial vegeta­tion. The animals can be captured by fishnet or by spe­cial traps located on the water surface, and marked with numbered plastic labels in order to recognize specimens with the aid of field glasses. An important

objective of monitoring activities should be the detec­tion of reproductive sites in order to include them in strictly protected areas. Reproductive sites are well ex­posed land parcels with scattered vegetation and sandy soils; sometimes, they are sand beaches located along a rivercourse or a lake shore, but in many cases they may be very distant from water. The detection of nesting sites requires a very expert personnel.

LIZARDS

Twenty-five species of lizards occur in the Italian fau­na. On the whole, lizards can be found in all the habitat types occurring in Italy, from coastal dunes to alpine grasslands. They are found in forests, ecotones, maquis, bushlands and grasslands, on all the kinds of substra­tum (sand, clay, litter, hard rocks, etc.), including hu­man settlements. They are mainly active at daytime (Lacertidae, Scincidae) but some species are partially nocturnal (Anguidae and Gekkonidae).

Both adults and juveniles can be detected at sight, when they are walking on stones, rocks, walls, and tree trunks or in the midst of grass cover. Weekly surveys along transects in suitable habitats provide an indication of adult numbers and permit the localization of their for­aging areas. The survey period, according to the species, may vary from early spring to early autumn, during sun­ny or rainy days, at day or nightime. The reproductive period can vary from early spring to summer months, according to the altitude above sea level. To detect spec­imens and to achieve a safe taxonomic identification, the workers can use field glasses (8 x 30, 10 x 50), especial­ly on rocky habitats or where more species of related lizards live together. Slow worms and skinks are usually found under large stones or fallen tree trunks; stones and trunks may be a refuge for all lizard species during occasional rainy days in the midst of summer.

In order to estimate the home range size, lizards can be marked and recaptured. Captures are made by a loop positioned on the top of a thin stick. Toe clipping must be avoided because it may reduce the running or climb­ing ability of lizards. Also the injection of microtranspon-ders could be a negative impact on the ability of these animals whose safety is based on fast running and agile climbing. The better technique is marking the animals with an indelible dye or by a common nail-enamel. This method does not assure a permanent marking because the lizard skin is periodically changed. The marked indi­viduals must be sexed, photographed and then released in the same spot where captured because all the lizards are strictly territorial. Population density is easily calcu­lated by the ratio between the number of individuals and the surface of the habitats suitable for the species in the study area. Data for such estimate may be gathered by counting all the specimens encountered along a lin­ear transect during monthly surveys.

Dietary studies are possible using faeces analysis or

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stomach flushing. The latter consists of the introduction of water into the stomach of the animal by a thin catheter, in order to flush out the food remains. Such method must consider the trophic activity of the animals according to their behaviourial ecology and weather conditions. Generally speaking, captures should be made some hours after they started the foraging activity, allow­ing the animals to catch a certain number of preys. On the other hand, captures should not be made too late, when the animals could have ceased their foraging activ­ity and be digesting. The workers have to make some preparatory observations in order to assess well the for­aging activity of the animals as this may vary according to local conditions and weather. In order to reduce stress and risk of manipulation, stomach flushing should be ex­ecuted under anaesthesia (the worker should learn by experience with larger animals before treating the small­er ones). Moreover, the operation should be executed as soon as possible, in order to avoid the deterioration of food remains by digestive enzymes.

A special case is given by the Kotschy's gekko (Cirto-dactylus kotschyt), which occurs only in a restricted area of Puglie region, and lives along stone-walls. Dif­ferently from the other Italian species of Gekkonidae, it is a diurnal species and can be censused like a wall lizard (Podarcis spp.) in springtime, but it becomes nocturnal in summer when daily temperatures are too high. Stone-walls are treated as linear transects and the operator has to count all the individuals detected along them. Females live together forming small harems (usu­ally 4) within the range of a dominant male. The latter occupies a wall tract of about 10 m and is well recog­nizable for morphological and ethological characters. The technique for calculating the population size of this species is based on male counts assuming that for each male recorded, 4 females occur in its range.

Another special case is Podarcis raffonei, an endemic species with an extremely reduced range. It lives in the Aeolian Islands, mainly on some islets uninhabited by man. This rare species is associated to rocky areas cov­ered by a scattered bushy vegetation. It shows two ac­tivity peaks in March-June and in September-October. Owing to hot and dry conditions, the species is not ac­tive in summer months from July to August. To estimate population size and density, surveys are random and last 4 hours. Marking must be avoided because it could harm these threatened populations.

SNAKES

Snakes can be found in all the habitat types occurring in Italy, from coastal evergreen maquis to alpine grass­lands. They are found in forests, ecotones, maquis, bushlands and grasslands. They are mainly active at daytime but some species become nocturnal in summer months, during the hottest period. In order to monitor snake populations, the surveys can be conducted using

the method named 'time-constrained searching' without a defined linear transect. In fact, these animals show an aggregate distribution and linear transect could pass out of their home range. Captures are made by hand or with the aid of a Y stick having a gum garnish on the tip. Vipers can be handled from the tail but not from the neck to avoid anatomic damage. The survey period, according to the species, may vary from early spring to early autumn, during sunny or rainy days, at day or nightime. The reproductive period can vary from early spring to summer months, according to the altitude above sea level. Some species, like smooth snakes {Coronella) are usually found under large stones or fall­en tree trunks; stones and trunks may be a refuge for other species, mainly the juveniles, during occasional rainy days in the midst of summer.

In order to estimate the home range size, snakes can be marked and recaptured. All the captured specimens are measured, sexed and marked by the method named 'ventral scale clipping' i.e. removing one or more ventral scales according to a conventional code). This method, widely used by many authors, allows to recognize forev­er all the marked individuals. Adult females can be pal­pated for assessing their reproductive status and the number of eggs or offspring. The snakes must be re­leased in the same spot where captured because they are strictly territorial. Dietary studies are possible through palpation and inducing the snake to throw up the prey.

A special case is given by the meadow viper (.Vipera ursinii), whose Italian range is reduced to some moun­tain areas of Central Apennines (from 1500 to 2300 m a.s.l.). The activity period of this viper is very short (from end-April to half-September). Surveys for moni­toring the species should be carried out in May-June in the special habitat where it lives, i.e. montane bushland dominated by juniper. The snake is usually detected at the base of these plants, half concealed by vegetation, in the warmest hours of the day (from 10.00 to 16.00).

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