Biology of <Emphasis Type="Italic">Cheirogaleus major</Emphasis> in a Littoral Rain Forest in Southeast Madagascar

Greater dwarf lemurs (Cheirogaleus major) are small nocturnal primates from the rain forests of eastern Madagascar. I investigated a population of Cheirogaleus major in a littoral rain forest of Southeast Madagascar during 2 rainy seasons to supplement the sparse information available for the species. I collected data on morphology, group composition, sleeping behavior, home range, and social organization via mark/recapture, radio telemetry, and focal individual observations. I identified 2 presumed family groups, and my data from radiotracking revealed a monogamous social organization. In each group, I found an adult pair and its presumed offspring sharing home ranges and sleeping sites together. I also observed gregarious behavior of group members during their nocturnal activity. I found no difference in body measurements between sexes, but body mass and tail circumference increased significantly from November to February, indicating a fatting period before hibernating.     

Comparative analysis of <Emphasis Type="Italic">iol</Emphasis> clusters in <Emphasis Type="Italic">Lactobacillus casei</Emphasis> strains

The present study was designed to expand genetic knowledge of myo -inositol (MI) metabolism in Lactobacillus casei. Twenty-four L. casei isolates of dairy origin were tested for the presence of iol cluster. PCR screening revealed eight strains encoded functions involved in MI utilization, of which one strain was able to use MI as carbon source. To gain a deeper understanding of the function of iol genes, four of the eight observed iol clusters were subjected to the full sequencing procedure. The results showed that the iol cluster was not a common feature among dairy L. casei strains. In addition, the four iol clusters were highly similar to one another in terms of sequence similarity and operon architecture. However, abundant polymorphisms that comprised a majority of synonymous mutations were detected throughout the full sequences. Three of them distributed among iolB, iolC, and iolT genes were found in linkage to MI-negative phenotype. Compared with other bacterial iol clusters, the iol cluster of L. casei showed a high similarity with that of Bacillus subtilis.     

Interactions Between Third Parties and Consortship Partners in Tonkean Macaques (<Emphasis Type="Italic">Macaca tonkeana</Emphasis>)

Sexual competition is potentially disruptive for the cohesion of social groups because stress and conflicts can extend to other group members. The displays and interactions of sexual partners are liable to influence the behavior of group-mates, which may need to observe them to anticipate possible consequences. We studied 2 captive groups of Tonkean macaques (Macaca tonkeana) to test whether group-mates pay more attention to consort partners, modify their activities and social interactions, and exhibit signs of stress during periods of sexual consortships. We found that group-mates approached the top-ranking male more frequently and were more frequently oriented toward the consort pair at the time of consortship than at other times. Group-mates spent less time sleeping, and devoted less time to manipulating the environment and more time to monitoring during consortship. This indicates that consortships may incur costs in individuals not involved in sexual competition. However, Tonkean macaques did not exhibit any signs of increased stress during consortship periods, as their rates of scratching and yawning did not differ between consortship and nonconsortship periods. This study shows that not only direct competitors but also other individuals monitor the behavior of sexual partners. It is likely that group-mates obtain information this way about ongoing action, and take decisions accordingly.     

Social electric signals in freely moving dyads of <Emphasis Type="Italic">Brachyhypopomus pinnicaudatus</Emphasis>

Brachyhypopomus pinnicaudatus (pulse-type weakly electric fish) is a gregarious species that displays reproductive behavior and agonistic encounters between males only during the breeding season. During social interactions, in addition to its basal electric organ discharge (EOD), fish emit social electric signals (SESs) in the contexts of reproduction and intrasexual aggression. We reproduced natural behavior in laboratory settings: SESs recorded in the field are indistinguishable from those observed in our experimental setup. SESs are nocturnal, change seasonally and exhibit sexual dimorphism. This study provides an exhaustive characterization and classification of SESs produced by males and females during the breeding season. In male–female dyads, males produce accelerations and chirps while females interrupt their EODs. The same SESs are observed in male–male dyads. We present a novel, thorough classification of male chirps into four independent types (A, B, C, and M) based on their duration and internal structure. The type M chirp is only observed in male–male dyads. Chirps and interruptions, both in male–female and male–male dyads, are emitted in bouts, which are also grouped throughout the night. Our data suggest the existence of a sophisticated electric dialog during reproductive and aggressive interaction whose precise timing and behavioral significance are being investigated.     

The first smile: spontaneous smiles in newborn Japanese macaques (<Emphasis Type="Italic">Macaca fuscata</Emphasis>)

Spontaneous smiles are facial movements that are characterized by lip corner raises that occur during irregular sleep or drowsiness without known external or internal causes. They are shown by human infants and infant chimpanzees. These smiles are considered to be the developmental origin of smiling and laughter. There are some case studies showing that spontaneous smiles occur in Japanese macaques. The goals of this study were to investigate whether newborn Japanese macaques show a considerable number of spontaneous smiles thus to examine the mechanism of them. Seven newborn Japanese macaques were observed in a room for an average of 44 min, and incidental sleeping situations were monitored twice. All seven participants showed spontaneous smiles at least once during the observation. They showed 8.29 spontaneous smiles in average (SD = 10.89; 58 smiles in total), all found in the state of REM sleep. Thirty-nine of the 58 smiles were produced on the left side of the mouth. These characteristics were similar to those of spontaneous smiles in human infants. This is the first evidence that macaques as well as hominoids show a considerable number of spontaneous smiles. These phenomena may facilitate the development of the zygomaticus major muscle, which is implicated in smiling-like facial expressions.     

Fitness aspects of transgenic <Emphasis Type="Italic">Aedes fluviatilis</Emphasis> mosquitoes expressing a <Emphasis Type="Italic">Plasmodium</Emphasis>-blocking molecule

Vector-born diseases cause millions of deaths every year globally. Alternatives for the control of diseases such as malaria and dengue fever are urgently needed and the use of transgenic mosquitoes that block parasite/virus is a sound strategy to be used within control programs. However, prior to use transgenic mosquitoes as control tools, it is important to study their fitness since different biological aspects might influence their ability to disseminate and compete with wild populations. We previously reported the construction of four transgenic Aedes fluviatilis mosquito lines expressing a Plasmodium- blocking molecule (mutated bee venom phospholipase A₂–mPLA₂). Presently we studied two aspects of their fitness: body size, that has been used as a fitness-related status, and the expression of major enzymes classes involved in the metabolism of xenobiotics, including insecticides. Body size analysis (recorded by geometric wing morphometrics) indicated that both male and female mosquitoes were larger than the non-transgenic counterparts, suggesting that this characteristic might have an impact on their overall fitness. By contrast, no significant difference in the activity of enzymes related to metabolic insecticide resistance was detected in transgenic mosquitoes. The implication on fitness advantage of these features, towards the implementation of this strategy, is further discussed.     

Population genetic structure of wild and farmed rusa deer (<Emphasis Type="Italic">Cervus timorensis russa</Emphasis>) in New-Caledonia inferred from polymorphic microsatellite loci

Historical records indicate that 12 rusa deer (Cervus timorensis russa) were introduced in New-Caledonia during the 1870s. We used eight polymorphic microsatellite DNA loci to assess the genetic differentiation and diversity of farmed and wild deer populations. Past genetic bottlenecks were detected in both sub-populations, although higher genetic diversity was maintained in farmed populations, probably due to the regular introduction of reproducers from wild populations and from other farms. The genetic structure of farmed and wild populations differed significantly. There was a significant isolation by distance for wild populations, whereas farmed populations were significantly differentiated between farms independently from their geographical proximity. Wild rusa deer consisted of small populations (with effective population sizes ranging between 7 and 19 individuals depending on the methods used), with a low parent–offspring dispersion range (0.20–2.02 km). Genetic tools and direct observations provided congruent estimates of dispersion and population sizes. We discuss the relevance of our results for management purposes.     

Paternal Care in <Emphasis Type="Italic">Propithecus verreauxi coquereli</Emphasis>

Alloparental behavior is documented for several anthropoid primates, but few researchers have investigated the extent or variability of such behavior in prosimians. We report results from a study of male-infant interactions in 2 groups of Coquerel’s sifaka (Propithecus verreauxi coquereli) at the Duke University Primate Center (DUPC). Both groups contained 1 adult pair, 2 juveniles, and a newborn. The adult males exhibited paternal behavior toward their offspring in the form of grooming and holding the infant, though males differed in the amount of time they spent engaged in these activities. Group differences in the proximity maintained between the infants’ parents suggest that the relationship between adult males and females may help account for the variation. The presence of juveniles appeared to diminish paternal behavior in the group exhibiting a higher overall rate of male-infant interaction.     

Mapping species differences for adventitious rooting in a <Emphasis Type="Italic">Corymbia torelliana</Emphasis> × <Emphasis Type="Italic">Corymbia citriodora</Emphasis> subspecies <Emphasis Type="Italic">variegata</Emphasis> hybrid

Quantitative trait loci (QTL) detection was carried out for adventitious rooting and associated propagation traits in a second-generation outbred Corymbia torelliana × Corymbia citriodora subspecies variegata hybrid family (n = 186). The parental species of this cross are divergent in their capacity to develop roots adventitiously on stem cuttings and their propensity to form lignotubers. For the ten traits studied, there was one or two QTL detected, with some QTL explaining large amounts of phenotypic variation (e.g. 66% for one QTL for percentage rooting), suggesting that major effects influence rooting in this cross. Collocation of QTL for many strongly genetically correlated rooting traits to a single region on linkage group 12 suggested pleiotropy. A three locus model was most parsimonious for linkage group 12, however, as differences in QTL position and lower genetic correlations suggested separate loci for each of the traits of shoot production and root initiation. Species differences were thought to be the major source of phenotypic variation for some rooting rate and root quality traits because of the major QTL effects and up to 59-fold larger homospecific deviations (attributed to species differences) relative to heterospecific deviations (attributed to standing variation within species) evident at some QTL for these traits. A large homospecific/heterospecific ratio at major QTL suggested that the gene action evident in one cross may be indicative of gene action more broadly in hybrids between these species for some traits.     

Combined evaluation of bovine tuberculosis in wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>) and red deer (<Emphasis Type="Italic">Cervus elaphus</Emphasis>) from Central-East Portugal

Bovine tuberculosis (bTB), widely considered a disease of livestock, can also affect large game species despite their poorly understood role in the epidemiology of the endemic state of the disease. The Idanha-a-Nova County, situated in the eastern region of central Portugal, is one of the most important regions for large game hunting activity. In order to investigate the role of large game and the presence of bTB in Idanha-a-Nova, 132 wild boar (Sus scrofa) and 339 red deer (Cervus elaphus) were inspected for bTB-compatible lesions (bTBCL) during the 2008–2009 hunting season. Tissue samples with bTBCL were collected and processed for bacteriological confirmation and spoligotyping of Mycobacterium bovis isolates. In wild boar, bTB lesions were single, mostly calcified (21/29, 72.4%) and situated in the mandibular lymph nodes. Lesions were more severe and largely non-calcified (36/40, 90%) in red deer. Wild boar presented a consistently higher bTB infection rate than the red deer; however, the difference was only significant in the southern B area. This area was significantly affected by bTB for both animal species (up to 72.7% in wild boar and 34.8% in red deer) and requires the urgent implementation of intervention measures to contain the disease. Wild boar may assume an important role in the emergence of new disease foci in distant areas that infected red deer cannot reach. Spoligotyping of M. bovis isolates grouped the strains into three clusters, revealing patterns common to wild boar, red deer, and also to local cattle, indicating possible transmission among different animal species.     

Synthesis of orthogonally protected (<Emphasis Type="Italic">3R</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)- and (<Emphasis Type="Italic">3S</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)-4,5-diamino-3-hydroxypentanoic acids

Summary.  The paper describes two methods of the synthesis of ethyl (3R,4S)- and (3S,4S)-4-[(benzyloxycarbonyl)amino]-5-[(tert-butyloxycarbonyl)amino]-3-hydroxypentanoates, useful for the syntheses of edeine analogs. Differently N-protected (S)-2,3-diaminopropanoic acid was used as a substrate in both procedures. The absolute configuration of newly generated asymmetric carbon atoms C-3 in β-hydroxy-γ,δ-diamino products was assigned by means of ¹H NMR spectroscopy after their transformation into corresponding piperidin-2-ones. Received May 24, 2002 Accepted October 10, 2002 Published online December 18, 2002 Acknowledgment The authors are indebted to the Faculty of Chemistry, Technical University of Gdańsk for financial support. Authors' address: Zbigniew Czajgucki, M. Sc., Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Technical University of Gdańsk, 11/12 Narutowicza St., 80-952 Gdańsk, Poland, Fax +48 58 347 11 44, E-mail: zmczaj@wp.pl     

Life cycle of <Emphasis Type="Italic">Aylax hypecoi</Emphasis> (<Emphasis Type="Italic">Insecta: Hymenoptera: Cynipidae</Emphasis>), a gall inducer on <Emphasis Type="Italic">Hypecoum</Emphasis> ssp. (<Emphasis Type="Italic">Papaveraceae</Emphasis>)

The life cycle and developmental stages of Aylax hypecoi (Trotter, 1913, Hymenoptera: Cynipidae: Aylacini) were studied in detail. Aylax hypecoi is known to induce galls in fruits of two Hypecoum species — H. imberbe and H. geslini (Papaveraceae) and the larva develops in host plant fruits. The morphology and development of egg, larva and pupa were investigated, which has previously not been done. The shape and size of terminal-instar larvae and associated galls are sex-specific. Overwintering stage, adult emergence and flying periods, and egg productivity were studied also.     

Pungent odor of the adult skipper butterfly <Emphasis Type="Italic">Erynnis</Emphasis><Emphasis Type="Italic">montanus</Emphasis> (Lepidoptera: Hesperiidae)

Adults of some butterfly species have odors, several components of which are known to have pheromonal or defensive functions. Little is known about the odors of hesperiid butterflies, however. Erynnis montanus (Hesperiidae), especially male adults, emit a pungent odor that is detectable by the human nose. Chemical analysis has revealed that crude extracts of wild individuals contained 10 volatile substances, of which docosane and heneicosane were the main components. Because males contain a significantly larger amount of p-cresol than females, this aromatic compound is characterized as a strong male odor. At the end of the adult occurrence period, p-cresol decreased substantially in each male. Benzothiazole, identified for the first time in lepidopteran adults, was present in both sexes in almost equal amounts. Among the 10 volatile substances detected in males, biased distribution in the wings rather than the body was observed for benzothiazole, heptanal, and p-cresol. Male adults have androconial organs in the costal part of the forewing on which benzothiazole and p-cresol tended to concentrate. However, these compounds were detected not only in other parts of the forewing but also in the hindwing and body, suggesting the presence of undiscovered scent-producing organs.     

<Emphasis Type="Italic">Asthenes dorbignyi</Emphasis> (Passeriformes: Furnariidae) host of <Emphasis Type="Italic">Argas neghmei</Emphasis> (Acari: Argasidae)

Argas neghmei Kohls & Hoogstraal is a Neotropical tick species parasite of poultry and man in the arid western of the southern cone of America in Argentina and Chile. Males, females and nymphs of an argasid were collected from 20 to 22 of June, 2007 in nests of Creamy-breasted Canastero, Asthenes dorbignyi (Reinchenbach) (Passeriformes: Furnariidae) at about 7 km S of La Poma (24°46′S, 66°12′W) and a male tick at 24°48′S, 66°10′W on Nov 27, 2008, Salta province, Argentina. Scanning electron microscope micrographies and sequences of 16S rDNA confirmed that the ticks found were A. neghmei. Development in breeding areas of A. dorbignyi is probably a source of infestation for animal premises and houses. However, the role of other unknown wild birds may also contribute to the invasion of A. neghmei in chicken houses or residences.     

The seasonal occurrence of endophytic fungus, <Emphasis Type="Italic">Mycosphaerella buna</Emphasis>, in Japanese beech, <Emphasis Type="Italic">Fagus crenata</Emphasis>

The seasonal occurrence of Mycosphaerella buna in leaves and contiguous organs of Fagus crenata was studied in a Japanese beech forest, Ibaraki, Japan, in 1998 and 1999. Mycosphaerella buna was not isolated from newly developed leaves in May, but it was isolated from asymptomatic leaves after June. The frequency of its occurrence gradually increased until leaffall. In contrast, M. buna was not isolated from overwintered buds, leaf petioles, or contiguous current-year twigs. The spermogonia and pseudothecia were observed in dead leaves after leaffall. The mature pseudothecia were found on dead leaves from May to July. The ascospores produced in the pseudothecia were suggested to infect newly developed leaves.Contribution no. 173, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

<Emphasis Type="Italic">In vitro</Emphasis> propagation and secondary metabolites production in wild germander (<Emphasis Type="Italic">Teucrium polium</Emphasis> L.)

A protocol for in vitro propagation of the wild germander (Teucrium polium L.) was developed. In vitro plants were developed from ex vitro axillary buds. Then, shoot tips were excised and established on Murashige and Skoog medium. Proliferation of shoots was tested with different levels of 6-furfurylaminopurin, 6-benzyladenine, or thiadiazuron. The highest proliferation of T. polium was obtained when 6-benzyladenine and 6-furfurylaminopurin were used at 2.0 and 1.6 mg l⁻¹, respectively. Thiadiazuron gave the lowest response for shoot proliferation. Rooting was experimented at different levels of Indol-3-butric acid, Indol-3-acetic acid, or 1-naphthaleneacetic acid. 1-Naphthaleneacetic was the only growth regulator which promoted root induction. Rooted plants were acclimatized successfully with 75% survival and grown in the greenhouse. In vitro- and in vivo-grown plants were analyzed for essential oil production. In vitro-grown T. polium on MS medium supplemented with 6-benzyladenine and 1-naphthaleneacetic gave higher oil yield than that grown on hormone-free Murashige and Skoog medium. In vivo (wild)-grown T. polium produced different oil yield when collected in different months (April and October). β-caryophyllene, used as a marker compound in the essential oil, was identified and quantified by gas chromatography (GC) analysis. Gas chromatography/mass (GC-MS) spectrometry analysis was also used to identify other components of in vitro cultures and to compare with in vivo-grown plants.     

Variability of esterase patterns in adult flies of the <Emphasis Type="Italic">saltans</Emphasis> species group of <Emphasis Type="Italic">Drosophila</Emphasis> (subgenus <Emphasis Type="Italic">Sophophora</Emphasis>)

Esterases are known for their involvement in several physiological processes and high degree of polymorphism, in many organisms. Such polymorphism has been used to characterize species and species groups and to study genetic changes occurred in their evolutionary history. In the present study, the esterase patterns of 19 strains from 10 species representative of the five subgroups of the saltans species group were analyzed using polyacrylamide gel electrophoresis and α- and β- naphthyl acetates as substrates. Fifty-one esterase bands were detected and classified as 31 α-esterases, 18 β-esterases and two α/β-esterases. On the basis of the inhibition patterns using Malathion and eserine sulfate, 34 bands were classified as carboxylesterases, 14 as acethylesterases and three as cholinesterases. Ten gene loci were tentatively established on the basis of data on band position in the gel, substrate preference and inhibition pattern. Twenty bands were species-specific, the remaining being shared by species from the same or different subgroups. Bands detected exclusively in males and bands with a different frequency or degree of expression between sexes were also detected. In the gels prepared for analysis of gene expression in the body parts (head, thorax and abdomen), the degree of expression of the β-esterases was higher in the thorax, while the α-esterases were expressed predominantly in the abdomen and thorax. A global view of the data available at present on the esterases of the species from the saltans group and their degree of polymorphism are presented, as well as the possibility of using some β-esterases, because of their characteristics in the gels, as markers for species identification.     

Effects of sugars and growth regulators on <Emphasis Type="Italic">in vitro</Emphasis> growth of <Emphasis Type="Italic">Dactylorhiza</Emphasis> species

The influence of sugars and growth regulators on shoot and root growth of Dactylorhiza species was studied under in vitro conditions. The seedling development was stimulated with the application of glucose and sucrose at concentration of 10 g dm⁻³ each. The improvement of shoot growth rate and shoot length was enhanced by cytokinins N ⁶-(2-isopentenyl)adenine or N ⁶-benzyladenine and their combination with auxin indolebutyric acid (IBA). The root growth rate and root length of seedlings increased in the presence of IBA and α-naphthaleneacetic acid. Individual Dactylorhiza species showed statistically significant differences in shoot and root development depending on sugar and growth regulator combinations.     

Social Influences on Group Membership in <Emphasis Type="Italic">Propithecus verreauxi verreauxi</Emphasis>

The size and composition of a social group can influence the reproductive success of its members. I examined the hypothesis that residents actively try to manipulate group size and composition in Verreaux’s sifaka (Propithecus verreauxi verreauxi). I collected behavioral data from 21 individuals in 5 social groups from November 2000 to March 2002 in the Kirindy Forest (C.F.P.F.) of western Madagascar. I investigated the affiliative and agonistic behaviors of resident males and females toward other group members to determine whether residents attempt to manipulate the costs and benefits of group membership. I also examined 3 cases of immigration attempts and 1 case of emigration. As expected, group members facilitated the residency of subordinate members of the opposite sex. For example, females sought out the subordinate males, responded to their calls, and even groomed them more. However, intrasexual relationships were also important for establishing or maintaining residency. The data, in conjunction with the observations of immigration and emigration events, suggest that resident individuals of both sexes attempt to influence group size and composition. I suggest that single–male groups are not the norm in Propithecus verreauxi verreauxi because females encourage, and males do not discourage, multiple males to reside in the group to increase group stability and to reduce the risk of infanticide.     

Response to human presence during nocturnal line transect surveys in fallow deer (<Emphasis Type="Italic">Dama dama</Emphasis>) and wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>)

The use of line transect methodology and portable thermal imaging for ground survey of wildlife should require a good knowledge of the behavioural response of the animals to the presence of an observer, in order to take into account the potential bias in density estimate caused by deviation from the assumption that distances are recorded at the initial position. We used ten fallow deer and eight wild boar fitted with radiocollars to investigate animals’ response during simulated nocturnal line transect surveys, carried out in a Mediterranean plain forest. The experiment consisted in radiolocating a focal animal before and after an observer walked a transect nearby (<100 m). Each transect line was followed using a Global Positioning System (GPS) navigator. We carried out a total of 64 trials on fallow deer and 57 on wild boar. Results showed that despite most of the animals moved significantly in response to the observer (mean ± standard error, wild boar—95.3 ± 10.0 m; fallow deer—149.6 ± 14.2 m), the flying patterns were different in the two species: the reaction of fallow deer turned out to be more intense and directional with respect to that showed by wild boar. Although these results sound explorative, the experiment attempted here, for the first time, is relevant for an appropriate design of nocturnal distance-sampling surveys and gives information about potential bias arising from animal’s behavioural response. We believe that these first results may foster more in-deep analyses which are now made possible with the adoption of GPS technology for animal location.