Effects of Ketamine hydrochloride on hematological and serum biochemical parameters in Callithrix jacchus

In the present study we have compared the hematological and hematochemical values obtained in adult males of Callithrix jacchus, before and after Ketamine Hydrochloride administration. The blood samples were obtained before and shortly after the Ketamine injection. The comparison between the values before and after treatment reveal no significant differences in most hematological and hematochemical parameters. Only AST and CK were significantly increased after Ketamine exposure, whereas GGT and hematocrit were decreased.
Values obtained from animals before Ketamine Hydrochloride administration are in general agreement with those reported previously by other investigators.

The influence of motivation and social context in the consumption of novel food by young common marmosets (Callithrix jacchus)

The aim of this study was to verify the hypothesis that young naive common marmosets (Callithrix jacchus) learn from experienced members of their family to add novel foods to their diet. The hypothesis was tested in two experiments involving four families, for a total of 21 individuals. In the first experiment the youngest pair of juveniles of each family, ranging from five to seven months of age, was initially presented to a novel food item for 10 minutes. Then the members of their family, already experienced with that particular food, were introduced in a adjacent cage for other 10 minutes. In this cage there was the same food item presented to the naive young. The prediction in this first experiment, was that the quantity of novel food eaten by the naive young would have been greater in the presence of the family-mates eating it in the adjacent cage, than when alone. The second experiment looked at the importance of physical interactions in the consumption of novel food. Also in this case the naive young of each family were left alone with a novel food item for 10 minutes. But after the first part of the session the rest of the family was introduced in the same cage as the naive young. The hypothesis here was the physical contact would have lead to the greater consumption of novel food, compared when alone, by the naive young. The naive young of each family were presented, in each of the two experiments, with a highly-palatable novel food, and a medium-palatable novel food, to look for possible effects of motivation on the consumption of novel food. A control condition, for both kinds of novel foods, regarded the naive subjects left alone with a novel food for two consecutive 10 min half-sessions. Data collected concerned a series of food-relates activities, including exploring, eating, food sharing. A preliminary analysis of the results suggests that the naive young consumed more novel foods during the second 10min half-session in both experiments, than during the correspondent period of time in the controls. Furthermore, there was no significant difference in the consumption of highly-palatable novel food between the two experiments, whereas the young consumed significantly more medium-palatable novel food in the second experiment, where physical contact was possible, than in the first experiment, where only visual proximity was possible. These results will be discussed arguing for the importance of both motivation and social facilitation for the consumption of novel foods by naive young.

The economy of Capuchin monkeys' search behaviour in small and large scale environments

In previous studies aimed at investigating the relationship between memory for a set of locations and the use of spatial strategies, exhaustive search tasks have been implemented using touch-sensitive computer monitors as apparatus and capuchin monkeys (Cebus apella) as subjects. This touch screen procedure requires the subjects to touch all the icons of a set presented on a monitor. Capuchins are free to select any trajectory (the serial selection of one icon after the other) through the set, and are permitted to touch icons already selected. Provided the search is exhaustive they receive a reward. Over the course of task practice monkeys spontaneously reduce the number of redundant moves (i.e. repeats on locations already selected) and show a tendency to exploit some constraints afforded by the spatial arrangement of the search space. These regulatory functions can be related, on the one hand, to the evaluation of costs (in terms of time and/or energy) associated to redundant moves, and, on the other, to the emergence of spatial "strategies" devised to keep track of the locations already explored without overloading the memory system of the organism.
In the present study, the ecological validity of the touch screen based procedure, was evaluated by the use of a search task in a square enclosure with sides of 3 metres. Four capuchin monkeys (individually tested) were presented with a set of small plastic containers hanging from the ceiling of the cage, each containing a hidden bait. This task more closely resembles a natural foraging situation than the touch screen procedure: the distance between the containers is much larger than that between the icons presented on the touch-screen, and negative feedback (the absence of reward) occurs for each redundant move, providing an additional source of feedback. Therefore this task has a more explicit ecological flavour, the costs associated to each redundant move are increased, and the subject has an additional source of feedback onto which regulate its behaviour. The number of exhaustive searches performed spontaneously by the monkeys to recover the baits and the amount of redundant moves were recorded.
Our results were similar to studies using the touch screen methodology. This suggests that touch screen based procedures produce behaviours which are an adequate model of those deployed in more ecologically valid contexts. However, monkeys tested in the large enclosure produced a lower percentage of redundant moves compared with those tested with the touch screen methodology. This suggests that monkeys regulate better their search behaviour when costs are added to redundant moves and more explicit feedback is provided.

Infant maltreatment in a captive colony of Cotton top tamarins (Saguinus oedipus oedipus)

Infant maltreatment is a burning, contemporary socio-psychological and ethological problem which concerns human being as well as several other non-human primates. The cotton top tamarin is an endangered species that experiences, in captivity, a high rate of infant abuse and neglect. In particular, at the New England Regional Primate Research Center, 680 (52%) newborns were killed or rejected by their parents between 1984 and 1993. In the present work an attempt is made to identify the most important factors underlying the abusive parenting behavior and to correlate general behavior and parental competence. This report is based on over 400 hours of focal and ad libitum observations among 56 adult cotton top tamarins and three months of intensive archival study of nursery, veterinary, animal care, genealogical and research records. The most important factors associated with the maltreatment phenomenon were proximate explanatory variables related to parental experience and colony conditions: adults rejected in their own infancy, with little parental experience, in bad health or suffering severe stresses abused more often their offspring than the others did. Other important factors were infant's health condition and an inherited component. Abusing parents did not suffer of evident anomalies in their general behavior; anyway, it was possible to discriminate between "bad" and "good" parents looking at some cues: "bad" parents spent more time resting, scanning outside the cage and in non-social behavior than "good" parents did.

What can be learned from crowding experiments

The traditional view of crowding can be summarised as follows:
When population density increases, aggression and violence will also increase. In the last few years evidence has been gathered to conclude that there is not such a simple relation between amount of space and level of aggression in primates. An alternative "active coping" model has been proposed based on the notion that primates exhibit capacities of behavioural adjustment to environmental changes. This model predicts that aggression may increase somewhat at high densities but that severe forms of aggression will be limited. Recent studies on macaques and chimpanzees that avoid potentially confounding variables, such as novelty of the social and/or physical environment, support this model and suggest an important lesson for managing captive colonies of primates: Only well-established groups of primates when crowded into familiar areas are able to cope with the reduced inter individual distances. The effectiveness of primates in coping with crowding under such conditions should not lead us, however, to conclude that they are relaxed under high densities as social tension does increase when inter individual distances are reduced.

(supported by NIH grants No. RR-00165 and R01-RR09797)

The well-being of Primates in captivity (20 min Video)

E. Visalberghi, Istituto di Psicologia C.N.R., via U. Aldrovandi 16/B 00197 Roma

Researchers should care for the animals they keep for their experiments in their laboratory. In this respect, because of their cognitive abilities, primates deserve special attention and their physical and psychological well being should be monitored and improved. We outline some spects of their behaviour which can be used as indexes of their wellfare and show ways to improve it by enriching their social and physical environment. Regulations (including the Italian ones) concerning the maintenance of primates in the laboratories are briefly described and critically evaluated.
(The above video is available upon request from the Authors)
 

Fossil Primates from Italy: new finds

A. Mottura
Dipartimento di Biologia Animale, Universita'di Torino

L. Rook
Dipartimento di Scienze della Terra, Universita'di Firenze

Primates occur in the Italian fossil record spanning since the Late Miocene until Middle Pleistocene. They belong to the two super families of Hominoidea and Cercopithecoidea. Both have been known in Italy since the last century; the hominoids are represented by the peculiar endemic species Oreopithecus bambolii, while cercopithecids belong to members of the subfamilies Colobinae (Mesopithecus) and Cercopithecinae (Macaca). In recent times a number of new finds allow to a better knowledge for all these taxa.

Oreopithecus
Oreopithecus bambolii is an endemic fossil primate whose precise affinities have been the subject of much debate ever since Gervais' description in 1872. The genus was described on a specimen recovered from the Late Miocene lignities of Monte Bamboli (Grosseto), where it is associated with a peculiar highly endemic fauna. A larger Oreopithecus sample was recovered from a lignite mine in the Baccinello basin (Grosseto), in a faunal assemblage referred as V1. In the same basin, three successive endemic mammal associations are later recognised, indicated as V0, V1 and V2 (in chronological order) respectively. Previous studies showed that the material from Monte Bamboli (where Oreopithecus is associated with a V2 fauna) is indistinguishable from the Baccinello V1 sample. Until recent times, however, a direct comparison of Oreopithecus from different faunas in the same basin has been impossible because it was absent from the Baccinello V2 sample. Some recent finds at Baccinello, from sites with V2 fauna, allowed to analyse the problem. The study of this material allows in fact to conclude that there is no evidence of any significant morphological trend in Oreopithecus through time.

Mesopithecus
It is a small to medium-sized colobine living from the Late Miocene until to Pliocene of southern and central Europe. The genus is generally known by two different species. The oldest is Mesopithecus pentelicus (Vallesian - Turolian; Late Miocene), the second and younger, is M. monspessulanus (Ruscinian to early Villafranchian: Early to Late Pliocene). In spite of the rarity of the Late Miocene Italian localities with fossil mammals, in all such known faunas however Mesopithecus recurs, although it is not abundantly represented. All these oldest remains can be provisionally referred to M. pentelicus, pending the results of the study in progress. The younger and smaller species, M. monspessulanus is recorded in Italy only from Villafranca d'Asti (early Villafranchian). From this locality a new specimen, which improves the overall sample for the latter rarest taxa, has been found recently.

Macaca
The fossil remains of this genus, excluding the sample from Capo Figari (Sardinia), were always fragmentary and mainly represented by skulls (isolated teeth and mandibles), the post-cranial material resulting rare. The fossils belonging to the genus Macaca recovered so far in our peninsula, have been divided into two species: Macaca florentina (cf. sylvana), in north and central Italy, and Macaca majori, an endemic dwarf species from Sardinia. The former is the only circum-mediterranean fossil Macaca which can be clearly distinguished from the actual Macaca sylvanus. The conservative characters of the genus, the scarcity and the fragmentary condition of the findings, make it difficult not only to well establish the number of taxa but also the punctual differences between the extant and extinct forms. More recently, the lignite deposit of Pietrafitta (Perugia), have revealed a rich association of vertebrate fauna attributable to the late Villafranchian (early Pleistocene). Among these finds, there is a certain number of Macaca remains belonging to at least eight individuals and also representing different skeletal parts, both post-cranial and dental parts. The biochronologically calibrated and more representative population is important in the study of problems concerning taxonomy, phylogeny and ecology of this species.

Phylogenetic Relationships Among Strepsirhini Based on Mitochondrial DNA Sequences

G. Ardito
Istituto di Antropologia, Universita'di Firenze, Italy

Y. Rumpler
Institut d'Embriologie, FacultŽ de Medecine
Universita'L. Pasteur, Strasbourg, France

The evolutionary relationships among the Strepsirhini are one of the most long-standing controversies in primatology. Systematics and phylogeny of this group has been analysed using morphological characters, fossil evidence, ecological/ethological data and chromosomal banding patterns without reaching a consensus view.
Here, a reconstruction of the evolutionary pattern has been attempted by comparing mitochondrial DNA sequences from the cytochrome-b and the small ribosomal subunit (12s rRNA) genes. This work represents the most extensive (7 families, 29 species) study of this taxonomic group conducted at the molecular level by direct sequencing amplified DNA fragments.
Phylogenetic analysis of variable nucleotide positions was performed by several inference methods (parsimony, weighted parsimony, neighbor-joining and maximum likelihood) to verify whether alternative topologies were supported by different tree-building methods.
The evolutionary trees obtained provide insights for (I) the taxonomic recognition of the infraorders Lemuriformes and Lorisiformes, (II) the ancestral separation of Daubentonia, (III) the branching pattern of Cheirogaleidae, Lepilemuridae, Lemuridae and Indriidae. Phylogenetic relationships among each lineage are also described. Moreover, we discuss the phylogenetic implications of comparing data from genes with different structural and functional properties and nucleotide substitution rates. Congruence between different approaches must be viewed as the best way to approach phylogenetic questions permitting to evaluate the reliability of different data sets and tree-building methods.

Non-human Primates genetic typing by polymerase chain reaction

M. Failli, R. Martini
Istituto "IRC srl - Prof. M. Fanfani, 50129 Firenze

DNA analysis and genetic typing in man can be achieved by several techniques: RFLP analysis and DNA Fingerprinting with DNA Probes, Minisatellites and Microsatellites analysis.
Non-human Primates genetics has been greatly improved by new DNA technologies.
DNA typing sequences abundantly described in man have, often, a concrete counterpart even in several non-human primates and can be used, with various efficaciousness in Paternity testing and Primate colony management.
Here we critically revise the various methodologies available in individual typing in order to define genetic variability and to perform Paternity testing in Primates conservation and in colony management projects.
Samples belonging to several Primates species (Galago, Macaca, Papio, Cercopithecus, Presbytis, Callithrix, Cebus and Gorilla) have been tested with a panel of mini and microsatellites generally used in Forensics.
All tests have been performed by PCR amplification of DNA obtained from different materials. The same samples have been also tentatively typed for HLADQA1 and HLADRB systems with the reverse dot-blot hybridization technique.
Tests performed have given information on the possible applications of Minisatellites testing in Old World Monkeys and in Gorilla, and these data partially confirm data we have previously discussed.
Genetic variability (and its possible application in primate conservation and colony management) has been also tested by MAAP techniques (Single primer random amplification PCR).
Samples have been kindly provided by: S. Tofanelli (Galago fixed cytogenetic material); C. Carlˆ (Macaca fascicularis, M. arctoides, Callithrix jacchus); P. Cavagna (Presbytis cristatus and Cercopithecus aethiops immortalised cells, Papio and Gorilla gorilla hairs).

In situ restriction enzymes-nick translation procedure: a method to detect intraspecific variability of constitutive heterochromatin in Cebus.

The karyotypes of all the species belonging to Cebus genus (Cebidae: Platyrrhinae) are characterised by a very large amount of constitutive heterochromatin with both centromeric and intercalar distribution. This component of the genome moreover, reveals a very high level of inter and intraspecific variability. Recently, molecular studies have shown that two different high repeated DNAs, designated CAP A and CAP B, are localised in the intercalar and centromeric heterochromatic regions respectively. These peculiarities make this genus a unique biological model to study the role(s) that the heterochromatin can play in the complex of divergence phenomena.
Differently from the standard C-banding, the cytomolecular techniques, based on the in situ restriction endonucleases-nick translation procedure, are able to show the qualitative differences within this class of genome. We applied this experimental protocol in two subspecies of Cebus apella, C.a. paraguayanus and C.a. xantosternos, digesting metaphase chromosomes from PBL with Hae III and Alu I and detecting the enzymatic activity both by nick translation techniques and Giemsa staining.
The banding pattern induced by Hae III shows a different activity among the centromeric and intercalar heterochromatin, according to molecular data. Moreover the Hae III pattern is different in the two subspecies analysed. This last result in particular seems to confirm the efficiency of the use of cytomolecular techniques to detect differences in heterochromatic satellite DNAs, also at intraspecific level.

Fluorescent in situ hybridisation of human 28S rDNA probes in Prosimians, Old World Monkeys and Apes

P. Cavagna*, P. Finelli**, B. Chiarelli*, M. Rocchi**
* Istituto di Antropologia, Universita'di Firenze, via del Proconsolo 12, 50122 Firenze
** Istituto di Genetica, Universita'di Bari, via Amendola 165/A, 70122 Bari

The results of non radioactive in situ hybridization obtained by human ribosomal 28s DNA and Ag-staining are presented. Species belonging to Prosimians, Old World monkeys and Apes have been analysed. For some of these there were no previous observations or definitive results obtained by in situ hybridisation.
Results show for Varecia variegata (Lemuridae) a location of NORs in three pairs of small acrocentric chromosomes during metaphase; a distribution of ribosomal DNA and ribonucleoprotein in few small centres appears during interphase (fibroblasts). For Galago senegalensis moholi (Galaginae) we observed a location of NORs in two pairs of metacentric chromosomes and one pair of acrocentric in telomeric position; a tendency to connect various NORs to form "transverse stripes" appears during interphase (fibroblasts). In Presbytis cristatus (Colobidae) we observed a pericentrometric location of NORs in one single pair of metacentric chromosomes (marked chromosome"). One or two nucleoli with adjacent zones of more condensed ribosomal DNA characterise the interphases (lymphocytes). The constant correspondence between single NORs pair and no more of one-two nucleoli have been observed also in Hylobates lar (Hylobatidae) and Cercopithecus aethiops (Cercopithecidae), that belong to groups distinct from each other and from Presbytis. In Pan troglodytes (Pongidae) and Homo we observed a common pattern of telomeric location on five pairs of acrocentric chromosomes; a common topological scheme in few nucleoli with little grains of condensed ribosomal DNA characterise the interphases (lymphocytes). In Pongo pygmaeus we observed a location of NORs on seven pairs of acrocentric chromosomes; a correspondent topology of rDNA is observed during interphase (lymphocytes). Among catarrhini Pongo appears as the species with more elevate number of NORs, both in absolute as well as in ratio with chromosome number.

The Nucleolus of Presbytis cristatus by Optical-Electronic Microscopy and Image Analysis

We present the results of a morphogical and structural description of nucleolus in Presbytis cristatus (Colobidae 2n=44), obtained from preparations of lymphoblastoid cells transformed by Epstein-Barr virus. This analysis has been particularly devoted to the characterisation of nucleolus in a specific cytogenetic model: a) location of a single NOR pair in a pericentromeric position, as a common feature of old world monkeys; b) high metabolic rate and mitotic activity that result in NORs activation; c) frequent association of NORs observed in metaphase (about 20%) as a result of participating of both NORs to form a single nucleolus during interphase. We have considered also the lack of structural observations about nucleolus in old word monkeys.
Classical cytogenetic techniques have been utilised for optical and fluorescent microscopy: Acridine Orange staining, Ag-staining, Quinacrin/Dapi staining. The techniques of Osmium Ammine staining, and by Uranil Acetate and Lead Citrate EDTA have been utilised for electron microscopy. The results enabled us to evaluate the fibrillar, granular and chromatinic component as well as ribonucleoprotein, DNA and RNA in the nucleolus. Image Analysis (by NIH image 1.57 for Apple computer) applied to EM observations enabled us to consider spatial relations, electronic image colors on densitometric levels and proportional estimates of the outlined structures.

Macaca sylvanus as biological indicator to monitor the Cedar forest of Morocco: the model works !

Mohammed Arahou, Mohammed Mouna
Institute Scientifique, Univ. Mohammed V, Rabat, Morocco

Last year meeting we presented here a theoretical model based on various aspects of the demography of the Macaca sylvanus population and we hypotized that it could be used to document the impact of livestock grazing in the forest. Therefore the progressive desertification of these last North African forest despite some resistance of a few colleagues, now no more with us, we decided to start a pilot project with a number of volunteers from the Universities of Padova, Parma, Milan, Florence and Rabat. We identified an area of 500 sqKm around Azrou, we plotted 17 representative transect of 10 linear Km each and we replicated them 5 times in the summer of 1994 and two in the autumn of the same year. We censused over 3000 monkeys, with an average density of 29 monkeys per sqKm. We identified over 50% of the whole forest with heavy indication of incipient desertification. Density as expected was high in refugee areas, well inside the forest, and low in the peripheral areas, heavily impacted. Males, were highly over-represented as expected in refugee areas. Infant natality was initially unexpectedly high, according to the model, in the summer, but the next censuses in the autumn indicated that infant mortality in the first six month of life is over 4/5 of the total population of infants, a dramatically high figure. From these data we build a few deterministic model of population extinction rates that indicate that if the situation will not change in the future years, the Macaca sylvanus population for the Middle Atlas, the last viable population of this species, will go extinct between 60 and 100 years. The theoretical model we presented last year therefore proved its power, and we intend to continue to use it to document the changes, in terms of impact of this region. More data will be presented on the possible solution to protect these animals and the forests that shelters them.

Climatic influences on mother-infant interactions in captive Japanese macaques

This study evaluated the effects of time of day, ambient temperature, and relative humidity on mother-infant interactions in captive Japanese macaques (Macaca fuscata). Sixty-one mother-infant pairs were observed during the first 12 weeks of the infant life. Temperature, but not time of day and relative humidity, was shown to influence mother-infant interactions. Lower temperatures were associated to increases in the role played by infants in maintaining contact with their mothers and in maternal rejection, but did not affect total time spent in ventroventral contact. These results are interpreted in light of the different thermoregulatory needs of mothers and infants, and highlight the existence of a previously neglected cause for mother-infant conflict. It is hypothesised that climatic variations can have both sort-term effects on social interactions and long-term effects on the development of the infant personality.

Preliminary observations on the behaviour of a preschool children group

Dutch ethologist Frans de Waal was the first one to realise that chimpanzees can make peace after an aggressive interaction. Since de Waal defined such a behaviour as reconciliation, many studies have been carried out on that topic, and at the moment reconciliation has been observed in more than 10 species of primates, both in captivity and in the wild. These data suggest that such a mechanism of restoration of a social relationship is very important and it could have evolved in most monkey's societies. Recently, some studies have been carried out on the human species too, with particular attention to preschool children. These studies provide a good starting point for a further investigations, however they do not consent a comparison of results neither inside the human species nor with other species of primates, because the methodology is different in any of such researches. The present work constitutes a preliminary phase of an ethological study on a group of children in a nursery school at Rosignano (Livorno, Italy). In such a work the same methods are used as in the studies on non-human primates. Preliminary observations are carried out twice a week during free-play hours (8 to 10 a.m.), in order to restrict to a minimum interferences by teachers. Data collection is performed by means of a fixed videocamera. First results provided an ethogram and the behavioural profiles of children. These profiles have been related to the children home environment which has been defined on the basis of information given by the teachers.

Capuchin monkeys' (Cebus apella) performance in a task requiring cooperation

R. Chalmeau
Laboratoire de Neurobiologie et Comportement Universita'Paul Sabatier 118, route de Narbonne, 31062 Toulouse, France

Cooperation is a powerful way of increasing an individual's success and it is present in many animal species. The aim of the present study is to examine the extent to which capuchin monkeys are able to act cooperatively and to understand the role of the individual they cooperate with.
Two groups of capuchin monkeys (N=5 and 6) were tested to assess their ability to solve a task (cooperation task) whose solution required pulling two handles which were too far from one another to be pulled by one monkey only. Before carrying out this experiment, each group was trained to pull a handle (Training phase 1) and to pull two handles (which were both in reach) simultaneously (Training phase 2) in order to get a reward. Nine subjects were successful in training phase 1, and 5 in training phase 2. When training or testing occurred all group members could have access to the area in which the task was presented.
In the cooperation experiment, seven subjects were successful, i.e. pulled one handle while a companion was pulling the second one. To attempt to elucidate how capuchins reach success, and to what extent they possess an understanding of the role of the companion further analyses were carried out. They suggest that social tolerance, as well as explorative and manipulative tendencies were the major factors accounting for success: solvers did not increase their pull actions when a partner was close to or at the other handle, i.e., when cooperation might occur. Capuchins' results contrast with those of chimpanzees faced with the same task (Chalmeau 1994). Chimpanzees were less successful than capuchins but they paid attention to the partner's presence at the handle and coordinated their own behaviour with that of the partner.

Hand preference in the use of a tool by tufted capuchins (Cebus apella)

Manual preferences were examined in a tool-using task in 6 adult capuchin monkeys (Cebus apella). The task consisted of a hanging Plexiglas tube filled in the middle with a sticky reward; to obtain the reward the subject must hold the tube with one hand and insert the stick inside the tube entrance with the other hand. Hand preference was assessed in four components of the task requiring different motor skills: picking up the stick, insert the stick into the tube, pushing the stick to obtain the food, taking the food from the floor. Preferences were weaker when subjects performed simple unimanual actions of picking up the stick or taking food: 2 subjects showed a significant preference for the left hand, 1 for the right hand, 3 with no occurrence. Preferences were more strongly expressed in the components of the task involving complementary bimanual actions: a right hand preference was noted in 4 subjects, a left-hand preference in 2 subjects. These results only partially support for Mac Neilage et al. (1987) theory for a left-hand bias for visually guided movement and a right-hand bias for more complex manipulation in monkeys.

POSTER SESSION

The Primate Gallery in the Natural History Museum of the Universita'of Pisa

The Primate collection in the Natural History Museum of the Universita'of Pisa consists of numerous specimens (131 stuffed and 195 osteological specimens) that represent all the families of the order and 35 of 59 genera.
The collection is extremely important because it includes very rare specimens such as Daubentonia madagascariensis, Indri indri, Brachyteles arachnoides and Nasalis larvatus. For this reason it has been decided to dedicate a whole gallery of the Museum to the Primates exhibition. The exhibition-room follows two different criteria. The first (systematic criterion) is based on explanations, within the glass showcases, that describe the specimen and the general character of its group. The second (thematic-comparative criterion) is based on the presence of a series of wall posters explaining common themes of the exhibited groups and showing their differences. The gallery also includes an exhibition of Scadentia order; in only one glass showcase two stuffed specimens of Tupaia genus are exhibited.

The Primatological collection of the Zoological Museum "La Specola", Universita'of Florence, Italy

* Istituto di Antropologia, Universita'di Firenze
** Museo di Storia Naturale, sez. Zoologia "La Specola", Universita'di Firenze

The primatological collection of the Zoological Museum in Florence is the largest in Italy, with a total number of specimens of 456. These include 258 skulls, 75 skeletons and isolated bones, 205 stuffed specimens, 49 skins and 19 specimens in alcohol. The following families (classified following Corbet and Hill, 1991) are represented (number of species and specimens in brackets): Cheirogaleidae (2 species and 2 specimens), Daubentoniidae (1,2), Indriidae (2,9), Lemuridae (6,22), Lorisidae (7,34), Tarsiidae (1,1), Callithrichidae (10,39), Cebidae (20,93), Cercopithecidae (33,189), Hylobatidae (5,13), Pongidae (3,32), Hominidae (1,20). A large number of specimens were acquired between the end of 19th century and the beginning of 20th century. The largest single collection is that from the Italian Biological Expedition to Somalia (1959-1964), with 33 specimens, and from the expedition of Nello Beccari to the English Guyana (1931-1932), with 27 specimens. Among the oldest collections is a group of 13 specimens donated by the Grand Duchess of Tuscany in 1845-1846. Two typus specimens are present: the holotypus of Cercopithecus boutourlinii, Giglioli, 1887 (now C. mitis boutourlinii) and the paralectotypus of Cercopithecus brazzae, Milne Edwards, 1886 (now C. neglectus).

The Primatological Research in Pisa

Homo sapiens belongs, in all respects, to the Primate order. For this reason primatology is both a valid and a necessary support in the understanding of the human species. Many famous anthropologists (Le Gros Clarke, Leakey, Washburn, for example) were conscious of the importance of primatology. Raffaello Parenti, founding the anthropological school of Pisa, gave attention to the primatological studies as a mean to enrich the knowledge of mankind place in nature. Thanks to his contribution, nowadays primatology in Pisa is vital and rich of initiatives, although the study of primates in our country has always been full of obstacles. Since '80, the anthropological school of Pisa has carried out an intensive research program on several aspects of the biology and behaviour of human and non-human primates. In this report the above mentioned program is shown, which has been developed around three main subjects: the dental microwear of Platyrrhini and Catarrhini, the taxonomy of Lemuroidea and the ethology of human and non-human primates. The last subject has been articulated in several fields: social behaviour of macaques and chimpanzees, parental competence in tamarins, ecology and behaviour of lemurs, maltreatment and reconciliation of children. This work gave the possibility to carry out 12 master theses and 3 doctoral dissertations, over 17 papers, a bibliography on sexual dimorphism in primates which had international diffusion. Moreover an Universita'course in Primatology for Biological Sciences started in 1992. At present, 2 master theses, a doctoral and a post-doctoral researches are in progress, focusing on the ethology of lemurs, Callitrichidae and children. Therefore, the primatological program in Pisa is trying to keep vital a very important field and, at the same time, to promote the diffusion of primatological studies, both on a professional and on a popular level.

Soluble proteins in the gustatory and olfactory systems of Primates

Paolo Pelosi
Istituto di Industrie Agrarie, Universita'degli Studi di Pisa

Soluble proteins, belonging to the superfamily of lipocalins, could act as carriers for chemical stimuli. At present, the physiological function of these proteins has not been understood. Also the occurrence of these proteins in different species of Primates is unknown; attempts to identify proteins of the same class in other animal species, using the available antibodies and cDNA probes has failed in several cases; it is not clear whether the expression of these proteins is limited only to some species or their aminoacid sequences can greatly differ between species.
As a part of our investigation on the occurrence and structure of these proteins across different species of Primates, we have identified, purified and partially sequenced two members of this class from the von Ebner glands of two species of non-human primates, Callithrix jacchus and Cercopithecus aethiops and one member of this class from the human nasal mucus.
In the absence of any functional test, such as ligand-binding activity, we have used the indication of a low molecular weight in denaturing conditions (17-20 kDa) as a first criterion to identify putative lipocalins.
These proteins were purified by classical chromatographic methods, such as ion-exchange and gel filtration. Samples of the purified proteins were then subjected to direct sequential degradation, to determine the aminoacid sequence of their N-terminal region. This information provided clear evidence that these proteins thus purified from the above reported species belonged to the lipocalin family. Molecular weights are 19 kDa in native as well as in denaturing conditions for these proteins and the protein purified from C. aethiops was used to raise polyclonal antibodies in the rabbit. The antiserum, thus obtained, is currently used for the identification and affinity purification of similar proteins from other species of Primates.

Heterochromatin of Eulemur coronatus: FRET analysis and in situ hybridisation

Karyotype evolution of the genus Eulemur (Malagasy lemur, Prosimii) is mainly based on mechanisms of Robertsonian translocation (Rumpler et al., 1989). E. coronatus is the sole species which underwent a significant increase of GC-rich centromeric heterochromatin (Ronchetti et al., 1993). The purpose of this work was a comparative cytochemical analysis, by FRET (Fluorescence Resonance Energy Transfer), a spectrofluorometric technique, and in situ hybridisation, of the heterochromatins of E. coronatus (ECO) and E. macaco (EMA), a species with a much lower amount of C-heterochromatin. In order to minimize differences due to different chromosome condensation, we utilized metaphase plates from the hybrid between ECO and EMA. In order to investigate the heterochromatin structure and relative interspersion of the AT and GC bp we utilised a quantitative evaluation of FRET, after staining of metaphase plates with the AT-specific Hoechst 33258 (donor) and propidium iodide, specific for total DNA (acceptor). The higher values of FRET efficiency for the heterochromatin of ECO seem to be due to a different interspersion of AT and GC along the DNA strands rather than major or minor condensation of chromatin, as showed results obtained after destructive deproteinizing of chromosomes with increasing molarities of NaCl. Experiments of "in situ" hybridisation were performed on ECO and EMA metaphases with some probes from ECO (ECOBamHI, ECOPvuII, ECOHaeIII) and with the telomeric probe (TTAGGG)n. Results showed, for all the ECO probes, a strong signal of hybridisation at the centromere of all chromosomes of ECO. On the contrary EMA appeared negative for ECOBamHI and ECOPvuII, while for ECOHaeIII showed weak signals of hybridisation on some chromosomes. These qualitative differences suggested a higher complexity of the heterochromatin of ECO than EMA, due perhaps to the presence of different families of satellite DNAs in ECO. Moreover, this different composition could justify different centromeric values of FRET between ECO and EMA, supporting the hypothesis of different AT/GC sequences between their heterochromatins. Regards the telomeric probe (TTAGGG)n, both ECO and EMA showed a strong positivity at all the centromeres. This centromeric localisation was emphasised in metacentric chromosomes of ECO. This fact could be due to fusion of acrocentric chromosomes of the ancestral specie, E. fulvus (EFU), with subsequent amplification of retained stretches of telomeric sequences.

An evolutionary hypothesis for the primate immunoglobulin heavy chain gamma genes

M. Del Pero, S. Crovella,
Dip. Biologia Animale, Universita'di Torino, Italy
A. Bottaro
Dip. Biologia Molecolare, Universita'di Siena, Italy and Howard Huges Medical Institute Boston, USA.

Four Immunoglobulin gamma heavy chain isotypes are present both in man and mouse. The functional and structural features of the different isotypes, as well as the mechanisms regulating their expression, share several analogies in the two species. However, the phylogenetic relationships between the genes are not known, because of the complex concerted evolution of the Ig multigene locus.
In this work, we present data obtained from Southern blot analysis of the gamma genes in several primate species, including prosimians (Lemur catta), New World Monkeys (Saguinus oedipus) and Old World Monkeys (Cercopithecus aethiops and Macaca fascicularis). DNA samples were digested with restriction endonucleases and hybridized with several probes from the human IgG genes.
The preliminary results obtained show that a single IgG gene exists in prosimians (Lemur) and probably in New World Monkeys (Saguinus). That gene underwent subsequent duplications during the Catharrine radiation, reaching the complex structure observed in the two Cercopithecinae species.
These data suggest that the functional IgG isotypes complexity was independently shaped during the primate and rodent evolution by common selective pressures, yielding similar properties in the two evolutionary lineages.

Highly Repeated DNA Sequences Analysis of Tupaia glis (Tupaiidae, Scandentia)

The phylogenetic relationships between tree-shrews (Tupaiidae) and primates have been a longly debated issue of primate evolution since the beginning of this century. Initially considered as unspecialized placental mammals of the order Insectivora, tree-shrews were then classified as members of the family Tupaiidae in the order Primates. More recently, some authors have proposed to place the tree-shrews in the separate order Scandentia.
Satellite DNA sequences have been shown to be good indicators of phylogenetic relationships among several primate species. To ascertain the taxonomic position of Tupaiidae with respect to primates and insectivores, highly repeated DNA sequences of Tupaia glis (Tupaiidae, Scandentia) have been therefore analysed with restriction endonucleases and southern blotting techniques. Five highly repeated DNA fragments have been isolated from T. glis and hybridised with genomic DNAs of several non-human primate species and one insectivore (E. europaeus).
The preliminary findings suggest that the isolated highly repeated DNA fragments of T. glis are specific alpha-satellite DNAs, probably newly evolved in tree-shrews, and absent both in primates and insectivores. Moreover, an ÔancestralÕ highly repeated DNA sequence which hybridised with all prosimians did not gave positive signals if used as probe against T. glis DNA.
The results so far obtained provide further taxonomic cues for the classification of the Tupaiidae in the order Scandentia.

DNA hair typing: a RAPD taxonomic reconstruction of Primates

Recently many efforts have been paid to study the molecular taxonomy and phylogeny of Primates; one of the major difficulty is represented by the DNA sample collection, especially for those species becoming day by day rare. Blood collection is therefore very hard to perform and it is necessary to find out other DNA sources. For this reason we decided to extract DNA from hair bulb.
In this study we analysed the DNA of about 40 species ranging from Lemur catta to Homo sapiens, using a PCR-based technique known as RAPD (Random Amplified Polymorphic DNA). We used 3 single primers and the amplification patterns, resolved on standard agarose gel, were analysed in order to obtain distance matrix. These matrix were then employed for generating the dendrograms, through the NTSYS package of cluster analysis.
The RAPD technique, together with DNA hair extraction, seems to provide an useful and very fast method to study the molecular taxonomic relationships among Primates species; we feel, although this is only a preliminary work, that RAPD could be important to find out specific DNA sequences, especially for those species so far not extensively studied at the molecular level.

Coinvolgimento della corteccia frontale nel controllo oculomotorio durante le fasi del sonno

C. Lucchetti
Istituto di Fisiologia, Universita'di Trieste

In uno studio inteso ad approfondire gli aspetti oculomotori a livello della corteccia frontale dorsomediale nella scimmia (Macaca fascicularis) abbiamo registrato l'attivitˆ elettrica di 340 neuroni, di cui 25 durante sia la fase di veglia che durante la fase di transizione veglia-sonno.
Gli animali, in accordo con la direttiva europea, erano condizionati ad eseguire sia compiti di saccade che di fissazione. Il movimento oculare veniva derivato con la tecnica del campo magnetico ed i potenziali d'azione erano registrati con i comuni sistemi elettrofisiologici. Si derivava inoltre l'attivitˆ elettromiografica dei muscoli nucali, per definire lo stato comportamentale. Tutti i segnali erano digitalizzati e memorizzati mediante sistema MAC-GWI SSII per ulteriori analisi.
Tutte le 25 cellule presentavano una attivitˆ fasica post-saccadica, selettiva spazialmente durante il compito di saccade. Durante la fase di dormiveglia, quando il movimento oculare da saccadico diveniva un movimento lento, la scarica si modificava in quanto da fasica diventava tonica e perdeva le caratteristiche di selettivitˆ spaziale, in quanto era indipendente dalla direzione del movimento lento.
Le ipotesi che si possono fare sono: 1) l'attivitˆ cellulare  espressione di una attivitˆ propiocettiva; 2) la scarica neuronale  espressione di una attivazione centrale che entra nei processi di genesi dei movimenti oculari durante le fasi di sonno. Pensare che un recettore periferico cambi la sua attivitˆ in rapporto allo stato comportamentale non  sostenibile, in quanto l'attivitˆ recettoriale non si modifica al variare dello stato eccitatorio centrale. Inoltre non  ancora ben definita la funzione di eventuali propiocettori nei muscoli extraoculari nel primate non umano. La seconda ipotesi sembra pi attendibile in quanto attivitˆ similari sono state giˆ scoperte a livello del collicolo superiore e delle strutture del mesencefalo. Il prevalente orientamento verticale dei movimenti lenti durante la fase di dormiveglia potrebbe essere la conseguenza di una ridotta eccitabilitˆ delle strutture coinvolte nella genesi dei movimenti orizzontali.

Ricerche finanziate dal MURST 40% 60%, e svolte con l'apporto del CNR-ENEA (Centro di Medicina Sperimentale).

Image analysis of fluorescent R banding for Presbytis cristatus chromosomes

Cytogenetic preparations obtained from lymphoblastoid cells transformed by Epstein-Barr virus of Presbytis cristatus (Colobidae "n=44) have been studied by quinacrine, DA/DAPI banding and R fluorescent banding. In particular the analysis concentrate on R bands induced by Chromomycin A3/Methyl green, a specific staining for GC/AT rich chromatinic blocks in the scaffold structure of the chromosome. Images of preparations have been digitalized by a CCD camera in close circuit or by direct scanning of photographic negatives at high optical resolution (800-1200 dpi); NIH Image 1.57 software for Apple computer has been utilised to analyse the images.
In particular evaluations of chromosomal profile and density as well as frequency distribution of densitometric levels have been performed. The problems of background exclusion, variability between chromatids and between homologous chromosomes in different position of the metaphase plate have been considered.
The possibility to study the same chromosome at different levels of condensation in different metaphase stages made it possible to analyse the densitometric variations in the GC/AT bands.

NOR heteromorphism in Macaca nemestrina nemestrina/ Macaca nemestrina leonina genealogy

Cytogenetic analysis of Macaca nemestrina nemestrina/Macaca nemestrina leonina blood samples from the same family group showed us an etheromorphic variant of the marked chromosome present in the offspring father-daughter. The etheromorphism observed consists in an unequal duplication of NOR due to the presence of a satellite band in the achromatic region. Cytogenetic analysis have been performed by Giemsa stain, C and G banding, Ag staining for NORs, and have been supported by morphometric evaluations and statistical tests. Chromosomal dimensions have been measured by Image Analysis software (NIH Image 1.57 for Apple computers), and images have been digitalyzed by scansion at 400 dpi of photographic prints; 25 metaphase plates have been analysed. Morphometric analysis showed a significant alteration of dimensional parameters among homologous chromosomes: in the variant chromosome we observed respectively a larger NOR band and a minor length of the "long arm" while the "short arm" resulted equal between the homologous chromosomes. We argue about the cause of this variation as a translocation or a paracentric inversion in agreement with data of interphase association of homologous chromosomes. Finally we have considered the potential effects on fertility and variations on the sex ratio of the offspring, as observed in human genealogies for analogous chromosomal variations.

Preliminary study of Varecia varecia and Lemur catta vocalizations.

A preliminary analysis of vocalizations emitted by 3 Lemurids: Lemur catta and Varecia variegata (V.v.variegata variegata andV.v.rubra) was carried out on subjects living in the Parco Natura Viva of Pastrengo (VR) Italy.
The Lemur catta group consists of 6 animals; 2 adults (one male and one female), 2 subadults (male and female) and 2 twins born on 9 march 1995. The Varecia variegata rubra group consists of 5 animals: 3 adults (1 male and 2 females) and two subadults of both sexes. The Varecia variegata variegata group consists in 4 animals: One adult male and one female, one young female and a baby male born in 1995. These lemurs live in contiguous cages each consisting of am outdoor and am indoor room, with suitable structures for the physical exercise of animals.
Vocalizations were recorded with a Sony TC-D5PRO and Marantz CP 130 tape recorder and with Sony CCD-TR55E videotape recorder, provided with a Sennheiser K3U+ME 88 microphone. The analysis have been performed with Canary 1.1.1 program on Macintosh IIci computer. Samples were obtained through a total of two hours of recording of observation during different times of the day, starting from february 1995 until today. Vocalizations were recorded from 2 - 3 m distance. These of baby Varecia variegata variegata were also recorded during the first hours of life by a microphone put in the nest.
During the 4 days of permanence in the nest of the mother and the baby 17 hours of observations were recorded; afterwards observations were continued in the same way as for the rest of the group.
The qualitative and quantitative study of vocalisations permitted to describe the diurnal and seasonal cycle of vocalizations and their frequency per time unit. A list of vocalisations was edited to analyze phonetics and semantic variability for further comparative study of the vocal emissions of both species. Categories of recorded vocalisations were: calls of intragroup and intergroup communication, alarming or startling vocal emission, mother-offspring (and vice versa) contact vocal emissions, subordination and mating vocalisations (the letter recoreded only in V.v.variegat).

* Dipartimento di Biologia Animale, Universita' di Torino.
** H.S.R. di Milano

Are individual differences present in infant calls of Lemur catta ?

Contact calls of adult Lemur catta exhibit individual differences in some acustic parameters (Macedonia, 1986). To study when and how these differences of vocalizations emerge, we performed a sonographic analysis of the infant calls emitted by five young born since 1993 in a colony of L. catta housed at the Zoological Garden "Cittˆ di Pistoia".
Calls produced in the first three months of life were recorded at different hours during daytime, using a Marantz tape-recorder connected with an Audiotecnica unidirezional microphone. Six calls for subjects where then converted through an Audiomedia analog-digital converter and then treated by Canary 1.1.1 (Cornell Laboratory of Ornitology, New York), both on Macintosh LCIII. The acustic parameters considered were: duration, delay to peak amplitude, starting frequency, peak frequency, ending frequency, highest frequency, and lowest frequency, all measured on fundamental.
One can observe that: (i) frequencies tend to decreasewith age; (ii) lowest and ending frequencies often correspond, as well as highest and starting ones. Infant calls show the frequency modulation of the 'early-high' adult contact call.
Between subjects variability do not significantly exceed within subjects variability for any of the parameters considered. It may be that (i) individual recognition has a lesser function for young than for adults; or (ii) individuality is reflected in acustic parameters that we have not considered; or (iii) our analysis is not sensitive enought to detect them.