The discovery of <Emphasis Type="Italic">Caligus</Emphasis><Emphasis Type="Italic">temnodontis</Emphasis> Brian, 1924 (Copepoda: Caligidae) from the bluefish <Emphasis Type="Italic">Pomatomus saltatrix</Emphasis> (Linnaeus) in the eastern Mediterranean Sea

The temnodontis variety of Caligus mauritanicus Brian, 1924 described by Brian (1924) is a valid species known only from a single host species, Pomatomus saltatrix (Linnaeus). New material of this species has been examined from the same host fish caught from Abuqir Bay, Alexandria (Egypt), from Iskenderun Bay (Turkey) and from off the coast of South Africa. Using this material, C. temnodontis Brian, 1924 is redescribed and compared with related species. It is most closely related to the Indo-Pacific species C. pagrosomi Yamaguti, 1939.     

Endoparasite spectrum of wild cats (<Emphasis Type="Italic">Felis silvestris</Emphasis> Schreber, 1777) and domestic cats (<Emphasis Type="Italic">Felis catus</Emphasis> L.) from the Eifel,Pfalz region and Saarland,Germany

Between 1993 and 2002, carcasses from 15 wild cats (Felis silvestris) and 17 domestic cats (Felis catus) from the Eifel region, Pfalz region and the Saarland were collected and examined for endoparasites. Most cats were road casualties (74%), some died from disease (14%), some were shot (3%), or some died of unknown reasons (9%). Three wild cats were too decomposed for parasitological examination. Endoparasites were recovered in 14 wild cats (n = 15) and 11 domestic cats (n = 17). A total of eight endoparasite species were found in wild cats and six in domestic cats. The nematodes Toxocara mystax and Toxascaris leonina and the cestode Taenia taeniaeformis were the most prevalent parasites. Other helminths detected were Capillaria aerophila, Capillaria feliscati, Capillaria plica and Mesocestoides litteratus. The spiruride Petrowospirura petrowi was detected in Germany for the first time. The parasite fauna was more diverse in male than in female cats indicating a male-biased parasitism in the wild cats.     

Muscular architecture of <Emphasis Type="Italic">Milnesium tardigradum </Emphasis>and <Emphasis Type="Italic">Hypsibius </Emphasis>sp. (Eutardigrada,Tardigrada) with some data on <Emphasis Type="Italic">Ramazottius oberhaeuseri</Emphasis>

The architecture of the musculature of the eutardigrade species Milnesium tardigradum Doyère, 1840, Hypsibius sp. and Ramazzottius oberhaeuseri (Doyère in Ann Sci Nat Zool Sér 2(14):269–369, 1840) is investigated by phalloidin staining and confocal laser scanning microscopy. There are methodological problems in staining eutardigrades due to physiological alterations under stress (anhydrobiosis) and due to penetration problems of the cuticle. It is helpful to fix specimens in the state of asphyxy, where animals are stretched following an oxygen shortage in their environment. The musculatures of all three species correspond in their general architecture, but differ in detail, such as in the number of muscles. All muscles are isolated muscle strands. There are on each body side two dorsal and one ventral muscle strands, in addition to a system of dorsoventral, lateral and lateroventral muscles. Seven median ventral attachment points give rise to dorsoventral, ventrolateral and appendage muscles. The appendages receive several muscles originating dorsally and ventrally. The number of muscles and the arrangement differ in each appendage. The fourth appendage shows the greatest differences with a far smaller number of muscles compared to other species. The musculature shows comparably few strict segmental patterns, for example, the musculature of each appendage differs from the other ones. By comparison with literature data on the same species and data of Macrobiotus hufelandi it can be shown that eutardigrades have a roughly comparable muscular architecture, but that there are several differences in detail. Dedicated to Professor Westheide on the occasion of his 70th birthday.     

Microanatomy of the trophosome region of <Emphasis Type="Italic">Paracatenula</Emphasis> cf. <Emphasis Type="Italic">polyhymnia</Emphasis> (Catenulida,Platyhelminthes) and its intracellular symbionts

Marine catenulid platyhelminths of the genus Paracatenula lack mouth, pharynx and gut. They live in a symbiosis with intracellular bacteria which are restricted to the body region posterior to the brain. The symbiont-housing cells (bacteriocytes) collectively form the trophosome tissue, which functionally replaces the digestive tract. It constitutes the largest part of the body and is the most important synapomorphy of this group. While some other features of the Paracatenula anatomy have already been analyzed, an in-depth analysis of the trophosome region was missing. Here, we identify and characterize the composition of the trophosome and its surrounding tissue by analyzing series of ultra-thin cross-sections of the species Paracatenula cf. polyhymnia. For the first time, a protonephridium is detected in a Paracatenula species, but it is morphologically reduced and most likely not functional. Cells containing needle-like inclusions in the reference species Paracatenula polyhymnia Sterrer and Rieger, 1974 were thought to be sperm, and the inclusions interpreted as the sperm nucleus. Our analysis of similar cells and their inclusions by EDX and Raman microspectroscopy documents an inorganic spicule consisting of a unique magnesium–phosphate compound. Furthermore, we identify the neoblast stem cells located underneath the epidermis. Except for the modifications due to the symbiotic lifestyle and the enigmatic spicule cells, the organization of Paracatenula cf. polyhymnia conforms to that of the Catenulida in all studied aspects. Therefore, this species represents an excellent model system for further studies of host adaptation to an obligate symbiotic lifestyle.     

Cold storage and cryopreservation of hairy root cultures of medicinal plant <Emphasis Type="Italic">Eruca sativa</Emphasis> Mill., <Emphasis Type="Italic">Astragalus membranaceus</Emphasis> and <Emphasis Type="Italic">Gentiana macrophylla</Emphasis> Pall.

To explore the possibility of an effectively long-term preservation of the germplasm of the HR lines of medicinal plant Astragalus membranaceus, Gentiana macrophylla Pall., and Eruca sativa Mill., both cold storage and cryopreservation approaches were attempted and compared. After 5-month cold storage on half strength Murashige and Skoog (1962) (1/2 MS) agar medium (AM), up to 82.9, 75.7, and 100% of the A. membranaceus, G. macrophylla and E. sativa hairy roots (HRs) recovered growth, respectively. The survival rates of A. membranaceus and G. macrophylla HRs significantly decreased, whereas that of E. sativa HR was unchanged with the addition of increased levels of exogenous abscisic acid (ABA) during cold storage. Using the encapsulation–vitrification (EV) method for cryopreservation, the G. macrophylla HRs died, whereas up to 6 and 73% of the A. membranaceus and E. sativa HRs survived, respectively. The HR lines evaluated with both methods showed no significant differences in morphology and growth rate compared with controls that were not subjected to preservation methods. These results suggest that cold storage is a more suitable alternative for the HR lines of the three studied plant species and that specificity of plant species have profound effects on the effectiveness of preservation.     

Identification of a major QTL for time of initial vegetative budbreak in apple (<Emphasis Type="Italic">Malus</Emphasis> x <Emphasis Type="Italic">domestica</Emphasis> Borkh.)

In the Western Cape region of South Africa, dormancy release and the onset of growth does not occur normally in apple (Malus x domestica Borkh.) trees during spring due to the mild winter conditions experienced and fluctuations in temperatures experienced during and between winters. In this region, the application of chemicals to induce the release of dormancy forms part of standard orchard management. Increasing awareness of the environmental impact of chemical sprays and global warming has led to the demand for new apple cultivars better adapted to local climatic conditions. We report the construction of framework genetic maps in two F1 crosses using the low chilling cultivar ‘Anna’ as common male parent and the higher chill requiring cultivars ‘Golden Delicious’ and ‘Sharpe’s Early’ as female parents. The maps were constructed using 320 simple sequence repeats, including 116 new markers developed from expressed sequence tags. These maps were used to identify quantitative trait loci (QTL) for time of initial vegetative budbreak (IVB), a dormancy related characteristic. Time of IVB was assessed four times over a 6-year period in ‘Golden Delicious’ x ‘Anna’ seedlings kept in seedling bags under shade in the nursery. The trait was assessed for 3 years on adult full-sib trees derived from a cross between ‘Sharpe’s Early’ and ‘Anna’ as well as for 3 years on replicates of these seedlings obtained by clonal propagation onto rootstocks. A single major QTL for time of IVB was identified on linkage group (LG) 9. This QTL remained consistent in different genetic backgrounds and at different developmental stages. The QTL may co-localize with a QTL for leaf break identified on LG 3 by Conner et al. (1998), a LG that was, after the implementation of transferable microsatellite markers, shown to be homologous to the LG now known to be LG 9 (Kenis and Keulemans 2004). These results contribute towards a better understanding regarding the genetic control of IVB in apple and will also be used to elucidate the genetic basis of other dormancy related traits such as time of initial reproductive budbreak and number of vegetative and reproductive budbreak.     

A partial skeleton of <Emphasis Type="Italic">Cetorhinus parvus</Emphasis> Leriche, 1910 (Chondrichthyes,Cetorhinidae) from the Oligocene of Germany

A partial skeleton of a shark was found in the Frauenweiler clay pit near Rauenberg (Oligocene, Rupelian; Baden-Württemberg, S. Germany). The shark is identified as Cetorhinus parvus Leriche 1910. One tooth, vertebrae, dermal denticles and most of the filter apparatus of the juvenile specimen are described and illustrated. A terminology is introduced for the gill raker elements of the genus Cetorhinus to compare C. parvus and C. maximus.     

Multiple host-ant use by the predatory social parasite <Emphasis Type="Italic">Phengaris</Emphasis> (=<Emphasis Type="Italic">Maculinea</Emphasis>) <Emphasis Type="Italic">arion</Emphasis> (Lepidoptera,Lycaenidae)

Phengaris (=Maculinea) arion is an endangered social parasite of Myrmica ants, and for a very long time was considered as specific to Myrmica sabuleti. Previous studies carried out in Poland suggested some discrepancies within this assumption, and therefore a much more intensive survey was undertaken. The host ant use of P. arion was studied at five sites in different types of biotopes in Poland, i.e. xerothermal grasslands where Thymus pulegioides was used as a larval food plant by the butterfly, and more or less sandy biotopes with Thymus serpyllum. Altogether nine Myrmica species were recorded, and considerable variation in species composition and density of nests was recorded. At four localities M. sabuleti proved to be the most common ant. A total of 529 Myrmica nests were examined, and only 20 of them contained larvae and pupae of P. arion. Host ants belonged to five different species, i.e. M. sabuleti, Myrmica scabrinodis, Myrmica schencki, Myrmica lobicornis and Myrmica hellenica. Only at one site (NE Poland) was a significant heterogeneity in parasitation rates among Myrmica species detected. M. lobicornis was the most often infested ant there, which may suggest local specialisation of the butterfly. Overall low parasitism rates may explain the vulnerability of P. arion in Central Europe but further studies are also necessary.     

Presence of <Emphasis Type="Italic">C. albidus</Emphasis>, <Emphasis Type="Italic">C. laurentii</Emphasis> and <Emphasis Type="Italic">C. uniguttulatus</Emphasis> in Crop and Droppings of Pigeon Lofts (<Emphasis Type="Italic">Columba livia</Emphasis>)

Columba livia is an important reservoir and carrier of Cryptococcus neoformans, Cryptococcus uniguttulatus, Cryptococcus laurentii and Cryptococcus albidus. Upper digestive tract of this species is also known as a habitat for Cryptococcus neoformans. Given the increasing clinical interest of this microorganism, 331 swabs from crop and 174 dropping samples from pigeon lofts in Grand Canary Island have been studied. The obtained results show an extensive presence samples 81 positive (24.47%) of Cryptococcus spp. in analysed crops: 32 (9.66%) for C. neoformans, 24 (7.2%) for C. uniguttulatus, 23 (6.9%) for C. albidus and 2 (0.6%) for C. laurentii. In the same way, Cryptococcus spp was also isolated in 82 (47.13%), dropping samples: C. neoformans in 59 (33.9%), C. uniguttulatus, in 9 (5.17%), C. laurentii in 8 (4.59%) and C. albidus in 6 (3.44%) of the investigated samples, respectively. The cryptococcosis produced by species of cryptococci other than C. neoformans has become more important during the last decade, supporting the study on the role of pigeon in the epidemiology of this disease.     

Chromosome numbers for three species of medicinal leeches (<Emphasis Type="Italic">Hirudo</Emphasis> spp.)

Karyological preparations were made from the testisacs of three medicinal leech species Hirudo medicinalis Linnaeus, 1758, H. verbana Carena, 1820 and H. orientalis Utevsky & Trontelj, 2005 . The samples originated from different populations in Azerbaijan, Kazakhstan, Russian Federation, Ukraine and Uzbekistan. The chromosome numbers were determined both from mitotic and meiotic stages of spermatogenesis using the propionic haematoxylin staining technique. All three species were found to have different haploid chromosome numbers: H. medicinalis 14, H. verbana 13 and H. orientalis 12, thus corroborating the validity of these taxa. The chromosomes can be classified as meta-, submetacentric and acrocentric. The chromosome numbers obtained are similar to that of the related species Haemopis sanguisuga (Linnaeus, 1758), which has 13 pairs.     

<Emphasis Type="Italic">Democricetodon</Emphasis> and <Emphasis Type="Italic">Megacricetodon</Emphasis> (Mammalia,Cricetidae) from the Miocene of Sandelzhausen,Southern Germany

The cricetid species Democricetodon gracilis (Fahlbusch, 1964), Democricetodon mutilus (Fahlbusch, 1964), Megacricetodon bavaricus (Fahlbusch, 1964), and Megacricetodon minor (Lartet, 1851) from Sandelzhausen (Early/Middle Miocene boundary, MN5, Southern Germany) are represented by more than 2,000 molars providing substantial data on the variability in sizes and on variation in morphologies. Temporal and spatial distributions of these species indicate that they are probably immigrants in SE Germany, originating from south-eastern or more southern areas. Except for M. minor, whose origin is not clear and could be regarded as an immigrant from more eastern areas. Using the length of the mesoloph in the second upper molars as an indicator of habitat, D. gracilis and M. minor had a preference for a humid habitat, D. mutilus and M. bavaricus a preference for drier habitats.      

Habitat,density, and spatial distribution of <Emphasis Type="Italic">Rivulus giarettai</Emphasis> (<Emphasis Type="Italic">Actinopterygii</Emphasis>, <Emphasis Type="Italic">Cyprinodontiformes</Emphasis>) in southeastern Brazil

We present data on the habitat, density, and spatial distribution of Rivulus giarettai, and discuss some biotic and abiotic variables related to its abundance in Free Flowing Waters (FFW) and Dam Reservoirs (DR) in palm grove (Mauritia flexuosa) marshes (Veredas) in Uberlandia, Minas Gerais, southeastern Brazil. The mean density (individuals/plot) of R. giarettai was about 13 times higher in FFW than in DR. In FFW, the density of R. giarettai was highest at intermediate amounts of substrate (plant mass) and it was positively rank-correlated with the depth, and the number of arthropods. Individuals occurred in an aggregated distribution. The aggregated pattern could be related to a concentration of individuals in microhabitats neither too much exposed nor completely saturated by plants. R. giarettai was relatively abundant and tolerant to slight man-made habitat modifications. Damming appeared to be especially problematic by negatively affecting its density.     

Photosynthetic and transpiration responses of <Emphasis Type="Italic">in vitro</Emphasis>-regenerated <Emphasis Type="Italic">Solanum nigrum</Emphasis> L. plants to <Emphasis Type="Italic">ex vitro</Emphasis> adaptation

In vitro regeneration of black nightshade (Solanum nigrum L.) plants was achieved through callus-mediated shoot organogenesis followed by 30 d indoor ex vitro adaptation to nutritional stress under environmental ambience and thereafter 6-d outdoor acclimatization in pots prior to field establishment. Relevant physiological parameters including pigment content, chlorophyll a fluorescence, net photosynthetic rate (P _N), transpiration rate (E), and stomatal conductance (g _s) of in vitro-regenerated plants were investigated during the course of ex vitro adaptation. During the first 4 d of indoor transplantation to potting substrate, there was a marginal reduction in the leaf chlorophyll and carotenoid contents but P _N and E were strongly reduced. The stomatal conductance and E/P _N ratio were significantly higher in plants up to 20 d of indoor adaptation than those of comparable age grown naturally from seeds. The shape of the OJIP fluorescence transient varied significantly with acclimatization, and the maximum change was observed at 2.0 ms. The 2.0 ms variable fluorescence (V _j), 30 ms relative fluorescence (M ₀), photon trapping probability (TR₀/Abs), and photosystem II (PSII) trapping rate (TR₀/RC) showed initial disturbance and subsequent stabilization during 30 d of indoor acclimatization. Energy dissipation (DI₀/RC) and electron transport probability (ET₀/TR₀) showed an initial phase of increase during the 4 d after plants were transplanted outdoors. During the 6-d outdoor acclimatization after transfer of plants to soil, no significant change in total chlorophylls and carotenoids, E, and g _s were observed, but P _N improved after reduction on the first d. The OJIP-derived parameters experienced change on the first d but were stabilized quickly thereafter. There was no significant difference between outdoor acclimatized plants and those of the seed-grown plants of comparable age with respect to photosynthetic and fluorescence parameters. Direct transfer of plants without indoor acclimatization, however, showed a completely different trend with respect to P _N, E, and OJIP fluorescence transients. The bearing of this study on optimizing micropropagation is discussed.     

Presence of <Emphasis Type="Italic">Ixodes neuquenensis</Emphasis> Ringuelet, 1947 (Acari: Ixodidae) on the endangered Neotropical marsupial Monito del monte (<Emphasis Type="Italic">Dromiciops gliroides</Emphasis> Thomas, 1894, Microbiotheria: Microbiotheriidae) at Chiloé Island,Chile

The presence of Ixodes neuquenensis (Ringuelet, Notas Mus la Plata 12:207–216, 1947) (Acari: Ixodidae) parasitizing populations of Dromiciops gliroides Thomas, 1894 (Microbiotheria: Microbiotheriidae) at Chiloé Island confirms that this tick species is established in Chile. No preference of the ticks for sex or age of the host was observed.     

Overexpression of the Chitosanase Gene in <Emphasis Type="Italic">Fusarium solani</Emphasis> via <Emphasis Type="Italic">Agrobacterium</Emphasis><Emphasis Type="Italic">tumefaciens</Emphasis>-Mediated Transformation

To overexpress the chitosanase gene (csn) in F. solani, a vector based on pCAMBIA 1300 was constructed. The csn gene, which is under control of the Aspergillus nidulans gpdA promoter and A. nidulans trpC terminator, was introduced back into the F. solani genome by Agrobacterium tumefaciens-mediated transformation, and the herbicide-resistance gene bar from Streptomyces hygroscopicus was used as the selection marker. Transformants which showed a significant increase in chitosanase production (~2.1-fold than control) were obtained. Southern blot analysis indicated that most transformants had a single-copy T-DNA integration.     

Isolation of <Emphasis Type="Italic">madA</Emphasis> homologs in <Emphasis Type="Italic">Pilobolus crystallinus</Emphasis>

Pilobolus crystallinus shows unique photoresponses at various growing stages. cDNAs for putative photoreceptors were cloned from this fungus. Three genes named pcmada1, pcmada2, and pcmada3 were identified from the PCR fragments, and amplified with degenerated primers for the LOV domain, which is conserved in many blue-light receptors. Deduced amino acid sequences for PCMADA1, PCMADA2, and PCMADA3 had one light-oxygen-voltage (LOV)-sensing and two PER-ARNT-SIM (PAS) domains. A zinc finger DNA-binding motif was conserved in the C-terminals of PCMADA1 and PCMADA3. However, PCMADA2 lacked the zinc finger motif. Expression of pcmada1 was suppressed by blue light whereas that of pcmada3 was promoted by blue-light irradiation.     

Gynogenesis in the vine cacti <Emphasis Type="Italic">Hylocereus</Emphasis> and <Emphasis Type="Italic">Selenicereus</Emphasis> (Cactaceae)

Gynogenesis was investigated on the allotetraploid Selenicereus megalanthus and the diploid Hylocereus polyrhizus and Hylocereus undatus vine cactus species. Unpollinated ovules from developing flower buds containing microspores at middle uninucleate developmental stage were cultured on MS basal medium containing 2,4-D/TDZ with different sucrose concentrations. Ovule size increased under dark culture conditions in all the three species and the level of response was species and sucrose concentration dependent. The best responses were achieved in the two S. megalanthus accessions, E-123 and J-80, at 0.18 and 0.26 M sucrose. Only ovule enlargement was obtained in H. undatus and both ovule enlargement and callus were obtained in H. polyrhizus. Development in both species ceased and embryoids were not formed. Plant regeneration was directly and indirectly obtained in both S. megalanthus accessions. Ploidy level was determined for a total of 29 S. megalanthus gynogenic plants using flow cytometry: 15 were found to be dihaploid (plants with the gametophytic chromosome number) and the other 14 were found to have higher ploidy levels. This is the first report of successful gynogenesis in Cactaceae. The dihaploids of S. megalanthus successfully produced by ovule culture techniques opens new perspectives in vine cacti breeding.     

<Emphasis Type="Italic">Bacillus anthracis</Emphasis>, <Emphasis Type="Italic">Francisella tularensis</Emphasis> and <Emphasis Type="Italic">Yersinia pestis</Emphasis>. The most important bacterial warfare agents — <Emphasis Type="Italic">review</Emphasis>

There are three most important bacterial causative agents of serious infections that could be misused for warfare purposes: Bacillus anthracis (the causative agent of anthrax) is the most frequently mentioned one; however, Fracisella tularensis (causing tularemia) and Yersinia pestis (the causative agent of plague) are further bacterial agents enlisted by Centers for Disease Control and Prevention into the category A of potential biological weapons. This review intends to summarize basic information about these bacterial agents. Military aspects of their pathogenesis and the detection techniques suitable for field use are discussed.     

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