The Intestinal Bacterial Community in the Food Waste-Reducing Larvae of <Emphasis Type="Italic">Hermetia illucens</Emphasis>

As it is known that food waste can be reduced by the larvae of Hermetia illucens (Black soldier fly, BSF), the scientific and commercial value of BSF larvae has increased recently. We hypothesised that the ability of catabolic degradation by BSF larvae might be due to intestinal microorganisms. Herein, we analysed the bacterial communities in the gut of BSF larvae by pyrosequencing of extracting intestinal metagenomic DNA from larvae that had been fed three different diets. The 16S rRNA sequencing results produced 9737, 9723 and 5985 PCR products from larval samples fed food waste, cooked rice and calf forage, respectively. A BLAST search using the EzTaxon program showed that the bacterial community in the gut of larvae fed three different diets was mainly composed of the four phyla with dissimilar proportions. Although the composition of the bacterial communities depended on the different nutrient sources, the identified bacterial strains in the gut of BSF larvae represented unique bacterial species that were unlike the intestinal microflora of other insects. Thus, our study analysed the structure of the bacterial communities in the gut of BSF larvae after three different feedings and assessed the application of particular bacteria for the efficient degradation of organic compounds.     

Melanin and Melanogenesis at Different Life Stages in <Emphasis Type="Italic">Hermetia illucens</Emphasis>

This work has demonstrated that the life stages of the Hermetia illucens fly are characterized by varying melanin contents with the maximum melanin amount being detected in the pupa. It has been observed that active synthesis of the pigment takes place in the prepupa and coincides with the time when the population migrated for pupation. It has been noted that a high manganese content in the prepupa and pupa indicates the important role that this biogenic element plays in organogenesis and melanogenesis in H. illucens. It has been revealed that in the imago and pupa, the melanin demonstrates broad-range antimicrobial activity and contribute to the insect protection against bacterial and fungal infections.     

Dispersal and Burial Behavior in Larvae of <Emphasis Type="Italic">Chrysomya megacephala</Emphasis> and <Emphasis Type="Italic">Chrysomya albiceps</Emphasis> (Diptera,Calliphoridae)

Blowflies use discrete and ephemeral substrates to feed their larva. After they run out of food, the larvae begin to disperse in order to find adequate places for pupation or additional food sources, a process named post-feeding larval dispersal. Briefly state the aspects and why they are important were studied in a circular arena of 25 cm in diameter and covered with wood shavings to a height of 40 cm allowing post-feeding dispersal from the center of the arena. Larvae of both Chrysomya albiceps and C. megacephala were used in five experiments for each species. For each pupa location, determined as distance from the center, depth, and weight were evaluated. Statistical tests were done to verify the relation between weight, depth and distance for pupation and for larvae of two species shows that the media distance is significantly different for two species and for C. megacephala this distance is greater than the distance for C. albiceps. The depth too is different for each species, as the larvae of C. megacephala buries deeper than C. albiceps. With relation of weight, there is no statistic evidence that have any difference between weights for pupation for each species.     

Evaluation of serum protein electrophoresis in greater rhea (<Emphasis Type="Italic">Rhea americana</Emphasis> Linnaeus, 1758)

Serum proteins from 47 healthy greater rheas (Rhea americana; male and female) were separated by electrophoresis in order to characterize normal reference ranges. Determination of total protein concentration was performed through biuret reaction. The mean value of total serum protein was 4.4 g/dL. Absolute concentrations of serum proteins were determined by agarose gel electrophoretic fractioning. Five fractions were analyzed: albumin, alpha, beta1, beta2, and gamma globulins, and the average values in grams per deciliter were 2.39, 0.32, 0.45, 0.30, and 0.79, respectively.     

Size-dependent foraging niches of European Perch <Emphasis Type="Italic">Perca fluviatilis</Emphasis> (Linnaeus, 1758) and North American Yellow Perch <Emphasis Type="Italic">Perca flavescens</Emphasis> (Mitchill, 1814)

Body size of consumer species is a fundamental trait that influences the trophic ecology of individuals and their contribution to the functioning of freshwater ecosystems. However, the relationship between body size and trophic ecology can be highly variable both within and between closely-related and similarly-sized species. In this study we compared the intra- and interspecific relationship between body size and trophic position for North American Yellow Perch Perca flavescens and European Perch Perca fluviatilis, which share similarities in morphology, life history traits and trophic requirements. We used stable isotope ratios (δ¹⁵N and δ¹³C) to characterize differences in size-dependency of trophic position and to trace consumer foraging history of Yellow Perch in lakes in the Northwestern United States and European Perch in lakes in Germany. The trophic position and stable isotope ratios of Yellow Perch and European Perch steadily increased with total body length, but European Perch were consistently feeding at higher trophic positions than Yellow Perch at a given length. European Perch occupied considerably higher trophic positions (mean trophic position = 3.9) than Yellow Perch (mean trophic position = 2.8). Large European Perch were increasingly piscivorous, whereas large Yellow Perch were more opportunistic and omnivorous predators of invertebrate prey. Overall, the trophic position among individual Yellow Perch varied more strongly than in European Perch. We conclude that both species similarly increase in trophic position with size, but the specific size-dependency of both trophic position and resource use varies with taxonomy and local ecological conditions. Thus, body size as a sole measure of trophic position should be considered cautiously when generalizing across populations and species.     

Distribution of pigments in the shell of the gastropod <Emphasis Type="Italic">Littorina obtusata</Emphasis> (Linnaeus, 1758)

The patterns of pigment distribution in the shell of the gastropod Littorina obtusata were studied. It was determined that the background coloration of the periwinkle shell resulted from the coloration interaction of the external and internal parts of the ostracum. The coloration of the latter can be monochromatic or twolayer. The external and/or internal zones of the ostracum can be colored purple, white, orange, yellow and greenish. Only 11 of 25 possible color combinations were recorded. Four additional versions of shell background coloration resulted from partial depigmentation of external or internal parts of ostracum. The mottling pattern corresponded to white or depigmented lenticular sites in the external part of the ostracum. The bands on the shell may be subdivided into two groups according to their means of formation. The bands that appeared due to redistribution of pigments forming background coloration belonged to the first group, the bands arising as a layer of additional (white, yellow, orange) pigment belonged to the second group. Various band versions could occur simultaneously in the same individual.     

Reproductive parameters of the common genet <Emphasis Type="Italic">Genetta genetta</Emphasis> (Linnaeus, 1758) in Southwest Europe

We used information from 148 litters belonging to 119 different wild and captive female common genets (Genetta genetta) in SW Europe over a period of almost 50 years to study certain reproductive parameters. Data were obtained from a combination of sources including carcasses, live captures, telemetry-based studies, captive genets, and reports in the literature. The circannual pattern of breeding was fairly similar in both wild and captive genets, with births occurring in all months except December, albeit with a large peak in spring and a secondary peak in autumn. The sex ratio at birth was near to 1:1, and mean litter size was 2.16 ± 0.76 cubs/litter with a range of 1–5 cubs. Litters of two cubs were the most frequent both in wild and captive settings. In captivity, we have observed the births of replacement litters and double litters; however, we did not find evidence of wild females that having two litters in the same year. We conclude that the possibility of breeding at any time of the year is facilitated by the continuous spermatogenesis of the males and by the ability of females to enter into estrus after losing a litter prematurely and to breed twice in the same year.     

A record of the Far Eastern crane fly <Emphasis Type="Italic">Tipula</Emphasis> (<Emphasis Type="Italic">Platytipula</Emphasis>) <Emphasis Type="Italic">moiwana</Emphasis> (Diptera,Tipulidae) in Moscow

The crane fly Tipula (Platytipula) moiwana (Matsumura, 1916) occurs in the southern Russian Far East, including Sakhalin Island and the Kuril Islands, in Japan (Hokkaido), and in northeastern China. Recently this species was found in Moscow. The reason for the appearance of this species in Moscow is unknown.     

Characteristics of lipid fractions of larvae of the black soldier fly <Emphasis Type="Italic">Hermetia illucens</Emphasis>

The lipid fraction of larvae of the black soldier fly Hermetia illucens was shown to contain lauric acid (38.43 wt %) and its esters, azelaic and sebacic acids, and azelaic acid dibutyl ester. The dominant compound in the group of identified glycerides was lauric acid monoglyceride (0.70 wt %). Glycerides were also represented by triglycerides and diglycerides of lauric acid. Sterols were represented primarily by phytosterols (over 75%), the major of which was alpha-sitosterol (45%). The identified lipid complex composition is apparently determined by the biological characteristics of the fly Hermetia illucens and ensures antibacterial defence of larvae and stability of lipids at changing ambient temperature.     

Biological Efficiency of the Prepupae <Emphasis Type="Italic">Hermetia illucens</Emphasis> in the Diet of the Young Mozambique Tilapia <Emphasis Type="Italic">Oreochromis mossambicus</Emphasis>

It has been noted that the introduction of small doses of dry prepupae of Hermetia illucens flies into the diet of the young Mozambique tilapia Oreochromis mossambicus (0.5 g/kg feed) had a positive effect on the growth and development of fish. It was found that the average daily gain in the experimental group was 0.7 g. It was established that the blood biochemical parameters were within the norm in the fish of all groups; however, in the experimental tilapia, the erythrocyte sedimentation rate was significantly lower and a tendency to increase in the amount of hemoglobin and glucose was noted. It was revealed that in the seminiferous tubules of fish in the experimental group, the number of mature sperm was 5.9 times higher compared with the control, indicating an increase in the rate of spermatogenesis. It is assumed that the observed effects are due to the high content of manganese in the prepupae present in a biologically active divalent form.     

<Emphasis Type="Italic">In silico</Emphasis> and <Emphasis Type="Italic">in situ</Emphasis> characterization of the zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) <Emphasis Type="Italic">gnrh3</Emphasis> (sGnRH) gene

Background

Gonadotropin releasing hormone (GnRH) is responsible for stimulation of gonadotropic hormone (GtH) in the hypothalamus-pituitary-gonadal axis (HPG). The regulatory mechanisms responsible for brain specificity make the promoter attractive for in silico analysis and reporter gene studies in zebrafish (Danio rerio).

Results

We have characterized a zebrafish [Trp⁷, Leu⁸] or salmon (s) GnRH variant, gnrh 3. The gene includes a 1.6 Kb upstream regulatory region and displays the conserved structure of 4 exons and 3 introns, as seen in other species. An in silico defined enhancer at -976 in the zebrafish promoter, containing adjacent binding sites for Oct-1, CREB and Sp1, was predicted in 2 mammalian and 5 teleost GnRH promoters. Reporter gene studies confirmed the importance of this enhancer for cell specific expression in zebrafish. Interestingly the promoter of human GnRH-I, known as mammalian GnRH (mGnRH), was shown capable of driving cell specific reporter gene expression in transgenic zebrafish.

Conclusions

The characterized zebrafish Gnrh3 decapeptide exhibits complete homology to the Atlantic salmon (Salmo salar) GnRH-III variant. In silico analysis of mammalian and teleost GnRH promoters revealed a conserved enhancer possessing binding sites for Oct-1, CREB and Sp1. Transgenic and transient reporter gene expression in zebrafish larvae, confirmed the importance of the in silico defined zebrafish enhancer at -976. The capability of the human GnRH-I promoter of directing cell specific reporter gene expression in zebrafish supports orthology between GnRH-I and GnRH-III.

     

Prevalence of agglutinating antibodies against<Emphasis Type="Italic"> Listeria monocytogenes</Emphasis> in chicks of the white stork (<Emphasis Type="Italic">Ciconia ciconia</Emphasis> Linnaeus, 1758) in Poland

In 2002 and 2003, blood samples from white stork (Ciconia ciconia) chicks were examined for the presence of antibodies against Listeria monocytogenes. Listeria antibodies were detected in 121 (59%) of 205 chick samples. The probability of Listeria antibodies being present increased with chick age; chicks detected with Listeria antibodies were in better condition than those without the bacterium.     

Molecular Detection of <Emphasis Type="Italic">Xenostrobus securis</Emphasis> and <Emphasis Type="Italic">Mytillus Galloprovincialis</Emphasis> Larvae in Galician Coast (Spain)

The mussel species Xenostrobus securis from New Zealand was detected in the Spanish coast recently, in the mouth of the Verdugo River into the Vigo Ria. In view of the great importance of the farm mussel sector in this region, the presence of this alien species greatly concerned producers and administration authorities, because of its potential medium- or long-term effects on the autochthonous species, Mytilus galloprovincialis, an important marine resource widely exploited in this location. The goal of this study was to develop a DNA-based technique to identify X. securis and M. galloprovincialis larvae in plankton samples, which would allow monitoring for the presence of X. securis in different points of the Vigo Ria. The techniques used were simplex and multiplex polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and fragment analysis. The application of this system to planktonic samples could be an effective means to assess the presence of the alien species, allowing monitoring if its dispersion is increasing, or on the contrary, if its distribution is restricted to the mouth of the Verdugo River, where X. securis was first detected. In addition, the application of this system at different times could be useful to assess the presence of larvae of these two species in the plankton.     

Isolation and characterization of the <Emphasis Type="Italic">Larix gmelinii </Emphasis><Emphasis Type="Italic">ANGUSTIFOLIA</Emphasis> (<Emphasis Type="Italic">LgAN</Emphasis>) gene

ANGUSTIFOLIA (AN), a plant homolog of C-terminal binding protein, controls the polar elongation of leaf cells and the trichome-branching pattern in Arabidopsis thaliana. In the present study, degenerate PCR was used to isolate an ortholog of AN, referred to as LgAN, from larch (Larix gmelinii). The LgAN cDNA is predicted to encode a protein of 646 amino acids that shows striking sequence similarity to AN proteins from other plants. The predicted amino acid sequence has a conserved NAD-dependent 2-hydroxy acid dehydrogenase (D2-HDH) motif and a plant AN-specific LxCxE/D motif at its N-terminus, as well as a plant-specific long C-terminal region. The LgAN gene is a single-copy gene that is expressed in all larch tissues. Expression of the LgAN cDNA rescued the leaf width and trichome-branching pattern defects in the angustifolia-1 (an-1) mutant of Arabidopsis, showing that the LgAN gene has effects complementary to those of AN. These results suggest that the LgAN gene has the same function as the AN gene.     

Replication timing of large <Emphasis Type="Italic">Sorex granarius</Emphasis> (<Emphasis Type="Italic">Soricidae</Emphasis>, <Emphasis Type="Italic">Eulipotyphla</Emphasis>) telomeres

Previously, we described the unique feature of telomeric regions in Iberian shrew Sorex granarius: its telomeres have two ranges of size, very small (3.8 kb of telomeric repeats on average) and very large discontinuous telomeres (213 kb) interrupted with 18S rDNA. In this study, we have demonstrated extraordinary replication pattern of S. granarius large telomeres that have not been shown before in other studied mammal. Using the ReD-FISH procedure, we observed prolonged, through S period, large telomere replication. Furthermore, revealed ReD-FISH asymmetric signals were probably caused by partial replication of telomeres within an hour of 5-bromodeoxyuridine treatment due to the large size and special organization. We also found that in contrast to the telomeric halo from primary fibroblasts of bovine, mink, and common shrew, telomere halo of S. granarius consists of multiple loops bundled together, some of which contain rDNA. Here, we suggested several replicons firing possibly stochastic in each large telomere. Finally, we performed the TIF assay to reveal DNA damage responses at the telomeres, and along with TIF in nuclei, we found large bodies of telomeric DNA and ?-H2AX in the cytoplasm and on the surface of fibroblasts. We discuss the possibility of additional origin activation together with recombination-dependent replication pathways, mainly homologous recombination including BIR for replication fork stagnation overcoming and further S. granarius large telomere replication.     

Subspecies of the Eeuropean beaver<Emphasis Type="Italic">Castor fiber</Emphasis> Linnaeus, 1758

Subspecific relationships of the European beaverCastor fiber have been obscured by failure to follow the rules of the International Code of Zoological Nomenclature and remain an open issue. Inconsequence in the use of subspecies names caused a nomenclatural as well as a taxonomic confusion. We discuss these controversial and incorrect nomenclatural decisions, and recognize 9 nominal subspecies of European beaver, bearing 9 available names:C. f. fiber Linnaeus, 1758,C. f. galliae Geoffroy, 1803, C. f. albicus Matschie, 1907,C. f. vistulanus Matschie, 1907,C. f. pohlei Serebrennikov, 1929,C. f. birulai Serebrennikov, 1929,C. f. tuvinicus Lavrov, 1969,C. f. belorussicus Lavrov, 1981, andC. f. orientoeuropaeus Lavrov, 1981.     

Hover-flies of the genus <Emphasis Type="Italic">Melanostoma</Emphasis> closely related to <Emphasis Type="Italic">Melanostoma dubium</Emphasis> (Diptera,Syrphidae)

Two new species of the genus Melanostoma Schin., 1860, closely related to M. dubium (Zetterstedt, 1838), are described, and a key to the species of this group is given. Melanostoma clausseni sp. n. and M. tschernovi sp. n. are closely related to M. dubium (Zett.), but differ in the uniformly black abdomen and legs. Melanostoma clausseni sp. n. differs from M. tschernovi sp. n. in the lustrous mesonotum and finely pruinose frons and face (in M. tschernovi, the face, frons, and mesonotum are covered with gray pruinosity). Holotypes of the new species are deposited in the collection of the Siberian Zoological Museum (Novosibirsk).     

Production of transgenic <Emphasis Type="Italic">Pinus armandii</Emphasis> plants harbouring <Emphasis Type="Italic">btCry</Emphasis>III(<Emphasis Type="Italic">A</Emphasis>) gene

A synthetic chimeric gene SbtCryIII(A) encoding the insecticidal protein btCryIII(A), was transformed into Pinus armandii embryos and embryogenic calli using Agrobacterium tumefaciens. Polymerase chain reaction and genomic DNA Southern blot analysis showed that the SbtCryIII(A) gene was integrated into the genome of transgenic Pinus armandii plants, and Northern blot analysis indicated that the SbtCryIII(A) gene was transcribed.