Assessment of natural and artificial propagation of the white-clawed crayfish (Austropotamobius pallipes species complex) in the Alpine region with nuclear and mitochondrial markers

In a joint analysis of nuclear (allozyme) and mitochondrial markers (sequence and restriction fragment length polymorphism (RFLP) analysis of the 12S and 16S genes), five genetically distinct groups of the Austropotamobius pallipes Lereboullet species complex were detected in the Alpine region. The geographical distribution of these evolutionary lineages coincided largely with several taxa (A.p. pallipes, A. berndhauseri, A.(p.) italicus) formerly defined on the basis of morphological characters. A low level of genetic variability was found within these lineages. For some populations, the combination of the two markers gave additional information about the likelihood of their natural or artificial origin. A hybrid zone was detected between A.p. pallipes and A. berndhauseri in the Lake Geneva area. The mosaic distribution of the five evolutionary lineages indicated that conservation efforts should be aimed at the level of local populations.     

Pollen concentrations in central Italy (Ascoli Piceno and Perugia)

Summary Air-borne pollen grains were collected using Hirst volumetric spore traps in both Ascoli Piceno and Perugia during the March–November 1983 period. Similar types of pollens were recorded in both towns. In Perugia, higher total amounts of Cupressaceae/Taxaceae pollens were recorded in March and of Quercus, Gramineae, Urticaceae and Oleaceae in May–June. However, the pollen concentration in the Ascoli Piceno atmosphere was higher in the August–October period owing to the pollen of the ?summer weeds?.

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Riassunto I granuli di polline aerodiffusi sono stati campionati con trappole volumetriche tipo Hirst sia in Ascoli Piceno che in Perugia durante il periodo marzo–novembre 1983. In entrambe le città sono stati rilevati tipi di polline simili. In Perugia sono state registrate concentrazioni di polline in atmosfera più elevate rispetto ad Ascoli Piceno in marzo per la pollinazione di Cupressaceae/Taxaceae e in maggio–giugno per la pollinazione di Quercus, Gramineae, Urticaceae e Oleaceae. In Ascoli Piceno, invece, la concentrazione di polline in atmosfera è state più elevata di Perugia nel periodo agosto–ottobre per la pollinazione delle ?erbe estive?.

     

Reproductive cycle of Pistacia lentiscus (Anacardiaceae) in Southern Italy

ABSTRACT

The reproductive cycle of Pistacia lentiscus L. from southern Italy is described with special emphasis on phenology and reproductive success. The species flowers in spring, its fruits ripen in autumn and has a latent ovary period during the summer. Vegetative growth starts immediately after anthesis (end of March) and finishes by the second week of June. Bud sections showed that flower induction occurred a few weeks after shoot development. However, no change in the overall appearance of the buds was noticeable before the following spring. Flowering is very synchronous within a plant, and a clear overlap in the flowering times of the two sexes occurred in the population studied. Although P. lentiscus seems to be highly reproductive because of the numerous fruits generally found on the plants in autumn, its reproductive success (as percent of flowers which develop fruits with seed) is low. Very large numbers of flowers never develop fruit and drop at different stages during fruit development. Moreover, numerous fruits have no seed because of parthenocarpy, embryo abortion or insect damage. In P. lentiscus, low reproductive success is not uniform within the population because the number of empty fruits per plant is highly variable between the plants. Our observations suggest that rain at flowering can make pollination a limiting factor for fruit set because of the reduction in the quantity of available pollen. Since these results are in agreement with those reported for populations in Israel and Spain, such phenomena could be genetically determined, and therefore common to this species throughout the Mediterranean region.     

Reproduction in an Irish lake population of the crayfish Austropotamobius pallipes (Lerehoullet)

SUMMARY. 1. Mature crayfish were collected in 1984 from an alkaline Irish lake before and after the mating season.
2. There was some correlation between frequency of mating and decrease in vasa deferentia weight in captive males, but in the field vasa deferentia weight losses were highly variable. The data indicate that one third of adult males may not have mated in the wild.
3. All females appear to have spawned, whether or not mated, with an average 85% reduction in ovary weights over the season.
4. Pleopodal egg counts directly after laboratory spawning were on average 81% of ovarian counts; in the field, an estimated 50 days after spawning, they were only 61% of estimated ovarian egg numbers.
5. Average ovarian egg size was positively correlated with female size. Also, larger crayfish had a wider range of egg sizes than smaller crayfish.
6. It is suggested that aggression and competition for shelter among brooding females may result in a progressive loss of eggs.     

Reproductive behavioral changes during the ovarian cycle of lesser bushbabies (Galago moholi) in captivity.

Behavioral patterns were quantified in seven heterosexual lesser bushbaby (Galago moholi) pairs during the estrous cycle to determine the relative significance of behavioral and nonbehavioral components of female sexuality in mate attraction. Increases in the occurrence of several male behaviors indicating female attractiveness were initiated during vaginal swelling when the female was sexually nonreceptive. Female behavioral estrus, as indicated by intromission, was confined to a portion of vaginal opening coinciding with proestrous and vaginal estrous smears. Female attractiveness was maintained for much of the period of vaginal opening, whereas female receptivity ended a day or two earlier than attractiveness. Female receptive and proceptive behaviors were not well defined or extensive, and few female behaviors exhibited significant changes during the cycle. Scent-marking behaviors, such as urine washes, and male grooms, were generally elevated outside the behavioral estrous period. In G. moholi, male sexual arousal appears to be elicited primarily by female attractiveness, while behavioral components of female sexuality appear to be less important in influencing mate attraction.     

Performance comparison among multivariate and data mining approaches to model presence/absence of Austropotamobius pallipes complex in Piedmont (North Western Italy)

Freshwater inhabitants in Piedmont (Italy) have been deeply disadvantaged by environmental changes caused by human disturbance. Hence there are engendered species that need human intervention of an entirely different kind – better management through the development of innovative practical tools. The most ecologically important of the river-dwelling invertebrates is a threatened species, the native white-clawed crayfish Austropotamobius pallipes. This is the species that we focused on in our effort to contribute to species conservation. Specifically we contrasted three different techniques of managing data relating to the presence/absence of this species: logistic regression, decision-tree models and artificial neural networks (ANN). Logistic regression and decision tree models (unpruned and pruned) performed worse than ANN. In this case, tree-pruning techniques did not make these models significantly more reliable, but did make the trees less complex and therefore did make the models clearer. ANN performed the best. Therefore we have judged them to be the most effective techniques.     

Histopathological survey of pathogens and commensals of white-clawed crayfish (Austropotamobius pallipes) in England and Wales

A histopathological survey of white-clawed crayfish (Austropotamobius pallipes) from six rivers in England and Wales was conducted between November 2007 and September 2011. A. pallipes bacilliform virus (ApBV) was present in four samples; infected crayfish showed pathological responses ranging from mild hypertrophy of infected nuclei and emargination of chromatin through to loss of architecture of the hepatopancreatic cells. Crayfish were found to be hosts to two different ciliate species, mites, nematodes, digeneans and the microsporidian Thelohania contejeani. The variation in disease presence between populations highlights the importance of conducting appropriate surveys of native crayfish prior to movement of animals to refugia sites for relocation, restocking or breeding programmes.     

Late Quaternary vegetation history at Stracciacappa (Rome, central Italy)

The pollen diagram from Stracciacappa (Sabatini volcanic complex, Rome) provides a record of vegetational and climatic change spanning the last 60000 years, which is the time since when volcanic activity in the crater came to an end. The chronological framework of the sediment core is set by five AMS and three conventional radiocarbon dates; the mean sedimentation rate obtained by radiocarbon measurements was used to extrapolate the age of the record beyond the reach of ¹⁴C dating. The sequence from Stracciacappa provides results of fundamental importance for the understanding of the vegetational changes which occurred during the last pleniglacial period in central Italy, and it can be considered as a reference pollen record for the regional biostratigraphic characterization of this period. The site shows a high climatic sensitivity, particularly highlighted by the development of some pleniglacial oscillations with woodland, which interrupted the succession of steppe and grassland vegetational formations typical of the glacial periods. Unfortunately, due to sedimentation problems and alteration of the top level sediments, the Holocene is only recorded in part. Only for one millennium, from ca. 8300 to 7200 uncalb.p., was a real forest expansion characterized by over 90% arboreal pollen found.     

Variation in the Ichnofauna of the Collio Formation (Lower Permian) in the South-Alpine Region (Northern Italy)

In the Collio Formation (Lower Permian) of the South-Apine region (North Italy) paleobiodiversity consists of tetrapod footprints, invertebrate traces and rarer plants, and is characterized by an absolute poverty of taxa that becomes more acute from the lower to the upper part of the Collio Formation. Regionally, the drop in diversity can be explained by joint tectonic and climatic change; humid evergreen environments became mainly dry. Examination of ichnofossil distribution supports this hypothesis.     

Metal-protein binding patterns in the hepatopancreas of the crayfish (Austropotamobius pallipes) during short term cadmium stress

Cd-binding patterns in the hepatopancreas of the crayfish (Austropotamobius pallipes) have been analysed in response to the administration of a single dose of Cd ions. Four main components, with apparent molecular weights of 3-4, 12, 20 and greater than 70 K daltons, have been isolated from soluble cell fractions. The variation in molar metal/protein ratios of these components is discussed in terms of the uptake, transfer and detoxification of Cd in the hepatopancreas. A model is proposed which describes Cd flux at the subcellular level and indicates that metallothionein-like proteins may mediate in this process.     

Diversity of mitochondrial DNA of the endangered white-clawed crayfish (Austropotamobius italicus) in the Po River catchment

1. The strong contraction of the European populations of white‐clawed crayfish has stimulated research of its population genetics. As part of a conservation project to introduce and improve Italian populations of Austropotamobius italicus, the genetic diversity of 16 populations from 10 tributaries of the Po River drainage was evaluated by sequencing a 494 bp length of cytochrome oxidase subunit I (COI mtDNA). 2. The evolutionary tree topologies and the parsimony network of 26 haplotypes separated into two clusters, on the basis of high average genetic distance (6.51 ± 0.42%). This finding indicated the presence of distinct evolutionary lineages, other than different allopatric sources. 3. An ancestral haplotype (Nav) was identified and 13 haplotypes, found mainly as singletons, were independently connected to Nav by short branches with one mutation site. Unimodal mismatch distribution and the goodness‐of‐fit of the observed data reflected a model of sudden population expansion, after the last glacial bottleneck event. 4. A general moderate level of genetic variability (h = 0.22–0.7 and π = 0.045–0.27%) was highlighted within A. italicus populations inhabiting Alpine and Apennine tributaries of the Po River. Analysis of molecular variance (amova ) revealed significant genetic structuring across all hierarchical levels, indicating that a major proportion of genetic variance is structured among localities within drainages. Multiple adjacent localities, strictly connected by the Po River, shared the two most frequent haplotypes (Nav and Laz), while remote populations, far from the Po River inflow, move towards a mtDNA fixation. 5. The results of this study indicate that distinct populations should be treated as separate entities for management. The key implication of the data is that ad hoc measures for successful programmes of recovery and management strategies for A. italicus conservation, such as the phylogeographic study of haplotype distribution at a local scale and the genetic variability of populations used as stock breeders, should be considered.     

Reproductive management of silver foxes (Vulpes vulpes) in captivity

Specific protocols need to be developed that take into consideration the requirements of silver foxes. This study was designed to investigate the reproduction of 26 pairs of silver foxes (Vulpes vulpes). Reproduction parameters (breeding season, age at puberty, fecundity and fertility) of the foxes were examined under experimental conditions, and new techniques to optimize fox breeding were assessed. Staining of the vaginal smears with the double Harris-Schorr stain allowed for the precise detection of estrus and proved to be more reliable than simply measuring vaginal resistance. Ultrasonography was demonstrated to be useful for pregnancy diagnosis and prediction of parturition. Results demonstrate that the methods reported in this paper are easily applicable to similar studies carried out on small groups of animals within the framework of fox animal experimentation.     

Role of the Dinaric Karst (western Balkans) in shaping the phylogeographic structure of the threatened crayfish Austropotamobius torrentium

1. This study examines phylogeography and phylogeny of the threatened stone crayfish, Austropotamobius torrentium, in order to elucidate the role of the Dinaric Karst geology in shaping the evolutionary history and genetic diversity of aquatic fauna in the western Balkans. Mitochondrial 16S rRNA and COI genes were partially sequenced from 188 and 159 crayfish, respectively, sampled from 70 localities. Phylogenetic relationships were reconstructed using four methods of phylogenetic inference. Divergence times between phylogroups were estimated in a Bayesian framework, and their demographic history was examined using neutrality tests and mismatch distribution analysis. 2. Seven geographically localised phylogroups separated by pronounced genetic gaps were found. Five of them have a distribution range within the northern‐central Dinaric (NCD) region, while the remaining two include populations from the southern Balkans (SB) and central and south‐eastern Europe (CSE). The oldest divergence event separated two NCD lineages from the rest of populations in the Late Miocene or Early Pliocene. Divergences amongst the five NCD phylogroups and SB + CSE occurred in the Pliocene. The most recent split separated SB and CSE phylogroups during the Late Pliocene. For both genes, uncorrected pairwise divergences between most of the phylogroups (4.1–8.7% for COI and 1.6–4.8% for 16S rRNA) were of the same range as, or higher than, some of the interspecific distances previously reported for the genus Austropotamobius. 3. Geographically isolated and deeply divergent cryptic monophyletic phylogroups within A. torrentium in the NCD region arose in the course of intensification of Neotectonic movements during the Pliocene and the beginning of the Pleistocene and the development of karstification that has heavily fragmented the palaeohydrography of the area. The results confirm a gradual north–south expansion of stone crayfish during the pre‐Pleistocene that preceded the rapid northward post‐glacial re/colonisation of central Europe (CSE phylogroup) through the Danube drainage. 4. Austropotamobius torrentium comprises morphologically cryptic but molecularly distinct taxa. Considering the relatively small geographical areas they inhabit, the NCD phylogroups of stone crayfish should be given the highest conservation priority.     

The reproductive performance of female Forest musk deer (Moschus berezovskii) in captivity

Reproductive performance of 750 adult female Forest musk deer (FMD, Moschus berezovskii) was monitored (from 2005 to 2009) on two Forest musk deer farms in Sichuan province, China. The mean (± SEM) lengths of the estrous cycle, pregnancy, and consecutive fawning intervals were 17.5 ± 0.8 d (n = 64), 182.9 ± 0.8 d (n =190), and 363.2 ± 2.4 d (n = 120), respectively. Copulation occurred from the end of October to early March, with the majority (84.1 ± 1.9%) during November and December (P < 0.05). The fawning rate was 68.7 ± 2.2% in yearling hinds and 73.3 ± 1.3% in adult hinds. Fawning occurred from the end of April to early September, with the majority (88. 9 ± 2.2%) during May and June (P < 0.005), including a peak in May (52.0 ± 1.0%). Yearling hinds fawned later (7.3 ± 1.8 d) than adult hinds. Rates of premature delivery and dystocia were 2.7 ± 0.4 and 2.0 ± 0.3%, respectively. There was a 1.07:1 female-to-male ratio at birth (P > 0.05), with 67.7% twins, but only two cases of triplets and one set of quadruplets among 537 fawns. Birth weight averaged 518.7 ± 10.3 g, ranging from 304 to 775 g (n = 240), with male fawns slightly heavier than females (521.1 ± 14.8 vs 516.9 ± 14.3 g). Postnatal and pre-weaning mortality rates were 25.0 ± 1.2 and 17.9 ± 3.1%, respectively. Postnatal mortality and urinary calculus were much more common in male fawns (P < 0.01). The mortality rate due to suppuration and dyspepsia were different (P < 0.001) between postnatal and pre-weaning periods. Due to higher mortality of male fawns, the adult sex ratio was significantly female-biased. In conclusion, captive FMD had similar seasonal patterns of copulation and fawning as wild FMD and other musk deer species.     

Introgression of chukar genes into a reintroduced red-legged partridge (Alectoris rufa) population in central Italy

Insight regarding the genetic origin and composition of the studied population of the red-legged partridge (Alectoris rufa) is likely to provide general and critical information for the appropriate management and possible conservation of the species. The reintroduced population of red-legged partridges living in Pianosa Island (National Park Tuscany Archipelago) has proven to be sustainable: captive-bred individuals, morphologically assigned to the taxon A. rufa, were released to the island approximately 20 years ago, establishing an apparently well-adapted population. We have investigated this population by means of 10 microsatellite loci in order to shed light on its genetic structure. Considering that A. rufa is known to crossbreed with A. chukar, the Pianosa Island population was compared at the molecular level with a red-legged partridge breeding stock (Aulla, MS) as well as with a population of pure A. chukar. Our results indicate that the red-legged partridge population from Pianosa, morphologically identified as A. rufa, is actually partly introgressed with A. chukar, questioning its genetic purity and the possible use of this population as a starting stock for future reintroductions elsewhere.     

Reproductive biology of female titi monkeys (Callicebus moloch) in captivity

Titi monkeys (Callicebus spp., Cebidae) are monogamous neotropical primates that live in family-like groups typically consisting of an adult monogamous pair and one or two young. Knowledge about the reproductive biology of this genus is scanty. This study investigated the reproductive biology of female dusky titi monkeys (Callicebus moloch). An initial analysis characterized reproductive parameters of 32 females from a captive colony maintained for 23 years at the California Regional Primate Research Center (CRPRC). The colony records provided data on reproductive parameters such as interbirth intervals, seasonality, age at first pregnancy, and reproductive rate in captivity. Changes in urinary levels of estrone conjugates (E1C) and pregnanediol-3alpha-glucuronide (PdG) were used to characterize major reproductive events. Urine samples from eleven females were collected during 17 months. The endocrine data were used to examine changes associated with cycling, conception, and the post-partum period as well as to determine the duration of the ovarian cycle and gestation length. The analysis of colony records indicated that females whose infant survived through weaning gave birth at intervals remarkably close to one year, while those who lost their offspring showed a significantly shorter interval. As long as they lived within the family group, mature female offspring did not breed. The analysis of the endocrine profiles indicated that after giving birth to a viable offspring, females undergo a relatively prolonged period of anovulation (approx. 6.5 months), followed by 1-3 non-conceptive cycles (approx. 1 month), after which they conceive and gestate (4.3 months).