Abstract: Monitoring wolves (Canis lupus) is a difficult and often expensive task due to high mobility,pack dynamic, shyness and nocturnal activity of this species. Wolves communicate acoustically trough howling, within pack and with packs of the neighbourhood. A wolf howl is a low frequency vocalization that can be transmitted over long distances and thus be used
for monitoring tasks. Animated howling survey is a current method to monitor wolves indifferent areas all over the world. Animated howling, however, may be invasive to residential wolf packs and could create possible negative reactions from local human population. Here we show that it is possible to detect wolves by recording spontaneous howling events. We measured the sound pressure level of wolf howls on captive individuals and we further found that simulated howling may be recorded and clearly identified up to a distance of 3 km. We finally conducted non-invasive acoustic detection of wolves in a free ranging population. The use of passive sound recorders may provide a powerful non-invasive tool for future wolf monitoring and thus help to established sustainable management plans for this species.

Abstract: 1. The recent recovery of the wolf in southern Europe has not yet removed the risk
of local extinction. Wolf populations are fragmented and often comprise fewer than
500 individuals. In North America, northern and eastern Europe, wolves feed maiiily
on wild herbivores. In southern Europe, this canid has apparently adapted to feed
also on fruit, rubbish, livestock, small and medium-size mammals.
2. The main conservation problem lies with predation o n domestic ~ingulates,w liich
leads to extensive killing of wolves. The reintroduction of wild large herbivores has
been advocated as a means of reducing attacks on livestock, but predatiori on the
latter may remain high if domestic ungulates are locally abundant.
3. Our synthesis of 15 studies, published in the last 15 years, on food habits of the
wolf in southern Europe, has shown that ungulates have been the main diet component
overall. A significant inverse correlation was found between the occurrence (%) of
wild and domestic ungulates in the diet. The presence of relatively few wild ungulate
species was necessary to reduce predation on livestock.
4. Selection of wild and domestic ungulate prey was influenced mainly by their local
abundance, but also by their accessibility. Feeding dependence on rubbish was local
and rare. In Italy, the consumption of riibbish/fruit and that of ungulates was significantly
negatively correlated. Diet breadth increased as the presence of large prey
in tlie diet decreased.
5. The simultaneous reintroduction of severa1 wild ungulate species is likely to reduce
predation on livestock and may prove to be one of the most effective conservation
measures.

Abstract: Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. The extraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artifi- cial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids. We recommend extracting faecal steroids by simply suspending the faeces in a high percentage of a primary alcohol (for glucocorticoid metabolites 80% aqueous methanol proved best suited for virtually all mammalian species tested so far). Not only does the procedure significantly increase the total amount of recovered radioactivity, it also increases the percentage of unconjugated metabolites, which are more likely to be recognized by the antibodies used in various immunoassays. The advantages of this extraction procedure are clear: it is very easy to use (no evaporation step is needed), it yields high recoveries and variation based on the extraction procedure is reduced to a minimum.