The diet of feral raccoon dog (<Emphasis Type="Italic">Nyctereutes procyonoides</Emphasis>) and native badger (<Emphasis Type="Italic">Meles meles</Emphasis>) and red fox (<Emphasis Type="Italic">Vulpes vulpes</Emphasis>) in Denmark

The raccoon dog (Nyctereutes procyonoides) is an East Asian Canid that has been introduced in Europe. Introduction of alien species is an increasing conservation issue. We examined the diet of a recently established raccoon dog population in Denmark by analysing stomach content in 249 carcasses collected in 2008–2016. Raccoon dog diet was compared to the diet of native badger (Meles meles) and red fox (Vulpes vulpes) in Denmark. The most common food for raccoon dogs were invertebrates (frequency of occurrence, FO 69%), small mammals (FO 68%), birds (FO 41%), fruits (FO 38%), amphibians (FO 36%) and carrions (FO 34%). The occurrence of invertebrates was highest during spring and summer, while fruits, cereals and carrions were eaten most often during autumn and winter. As expected, raccoon dog shared the major food categories with badger and red fox, but generally, it had a wider dietary niche. Overall, dietary overlap between raccoon dog and badger was 0.74 (Pianka index, O_jk). The dietary overlap with red fox was relatively high in all seasons, peaking in summer (O_jk 0.87) and dropping in winter (O_jk 0.79). Despite the dietary overlap between the alien racoon dog and native red fox and badger, the species may coexist due to partitioning of feeding habitats and/or because the red fox is limited by other factors, e.g. diseases and anthropogenic activities. The introduced raccoon dog seems to fit a dietary niche between badger and red foxes in human-dominated landscapes in north-western Europe.     

Transport pathways shape the biogeography of alien freshwater fishes in Australia

Aim

Changing preferences regarding which species humans have transported to new regions can have major consequences for the potential distribution of alien taxa, but the mechanisms shaping these patterns are poorly understood. We assessed the extent to which changes in human preferences for transporting and introducing alien freshwater fishes have altered their biogeography.

Location

Australia.

Methods

We compiled an up‐to‐date database of alien freshwater fishes established in drainages in Australia before and after the number of established alien fish species doubled (pre‐1970 and post‐1970, respectively). Using metacommunity models, we analysed the influence of species traits and drainage features on the distribution of alien fishes that established pre‐ and post‐1970.

Results

Alien fishes in Australia were introduced via four main transport pathways: acclimatization, aquaculture, biocontrol and ornamental trade. The relative importance of each pathway changed substantially between the two periods, accompanied by changes in the distribution of alien fishes and the variables predicting their distribution. Pre‐1970, most species (64%) were introduced by acclimatization societies for purposes such as angling and biocontrol, and these fish have established in inland drainages more heavily impacted by human activities. In contrast, most of the post‐1970 introductions (69%) were ornamental fishes, with most species established in small, north‐eastern, tropical and subtropical coastal drainages.

Main conclusions

Substantial changes in introduction preferences and transport pathways over time have altered both the patterns and underlying processes shaping the biogeography of alien fishes in Australia. Our findings highlight the need for caution when using historical data to infer potential future distributions of alien species. The continuing spread of alien species means traditional biogeographical units may no longer be identifiable in the foreseeable future.

     

Studies of the mechanism of bacterial resistance to complement-mediated killing. V. IgG and F(ab')2 mediate killing of E. coli 0111B4 by the alternative complement pathway without increasing C5b-9 deposition

The mechanism of antibody-dependent complement-(C) mediated killing of Escherichia coli 0111B4, strain 12015 (12015), was examined. 12015 was resistant to serum killing when incubated in hypogammaglobulinemic serum (H gamma S) or pooled normal human serum (NHS) that had been previously adsorbed to remove specific antibody (Abs NHS). Presensitization with immune rabbit serum or purified immune rabbit IgG resulted in 1 to 3 log killing when 5 X 10(8) colony forming units (CFU)/ml were incubated in 10 to 40% Abs NHS. Binding of 125I-C3 and 131I-C9 to the bacterial surface of the presensitized and the nonpresensitized strain was quantitated when these organisms were incubated in 10, 20, and 40% Abs NHS. Stable binding of up to 3.0 X 10(5) molecules of C3 and 8.0 X 10(4) molecules of C9 to presensitized and nonpresensitized isolates occurred in the highest concentration of serum, but there was no killing without presensitization. Similar results were found when Abs NHS was chelated with ethylene bis glycoltetraacetic acid containing 2 mM MgCl2 (Mg EGTA) to block classical pathway activation, indicating that antibody mediated the bactericidal reaction through the alternative pathway. Deposition of C3 and C9 and killing of 120 15 in 10% Abs NHS or 10% H gamma S was measured after presensitization with increasing amounts of IgG, F(ab')2, or Fab'. There was a dose-dependent increase in C3 deposition and killing, but only minimal change in C9 binding when 1.0 X 10(3) to 3.2 X 10(4) IgG or F(ab')2/CFU were bound to the bacterial surface. In contrast, there was no increase in C3 or C9 binding and no bacterial killing when 1 X 10(3) to 3.4 X 10(4) molecules Fab'/CFU were bound to the bacterial surface. These experiments show that immune IgG and F(ab')2 can mediate killing of E. Coli 0111B4 by the alternative pathway without changing the extent of terminal C component attachment to the bacterial surface.     

Transport and distribution of homocarnosine after intracerebroventricular and intravenous injection in the rat

Rats were injected intracerebroventricularly (i.c.v.) or i.v. with [¹⁴C]homocarnosine (250 nmol). Distribution of the dipeptide in brain structures, transport from the brain to the blood, distribution in peripheral organs, and excretion in the urine were studied by measuring radioactivity in tissue, plasma, and urine samples by liquid scintillation counting 15–120 min after injection. After i.c.v. injection, [¹⁴C]homocarnosine was taken up into all parts of the brain investigated (highest uptake in structures close to the site of injection), it was transported to the blood, and radioactive substances were found in low concentration in muscle, spleen, and liver, in high concentration in the kidneys, and very high concentration in the urine. Investigations using high pressure liquid chromatography (HPLC) showed that no degradation took place in the brain, all radioactivity was found in the homocarnosine fraction. In the plasma 86% of the radioactivity was found in the GABA fraction presumed to be formed by cleavage of the peptide, while in the kidneys 35% and in the urine 40% was found in the GABA fraction. After i.v. injection of [¹⁴C]homocarnosine, no radioactivity was measured in hippocampus, striatum, cerebellum and cerebral cortex 15 min after injection, however, 60 min after injection a very low activity was detected in these structures (estimated intravascular radioactivity subtracted). A low activity was also measured in the spinal cord both 15 and 60 min after injection. When homocarnosine and GABA were separated on HPLC, all radioactivity in brain tissue was found in the GABA fraction, indicating either that [¹⁴C]homocarnosine did not cross the blood-brain barrier in amounts that could be measured with the method used, or that peptide entering the brain was rapidly transported back to the blood. [¹⁴C]Homocarnosine was not taken up either into crude synaptosomal preparations from hippocampus, striatum, cerebellum, cortex and spinal cord, or into slices prepared from the hippocampus and striatum. Transport from the brain to the kidneys and excretion in the urine seems to be a major route for disposal of this peptide in the rat.     

Purification and characterization of a myosin I heavy chain kinase from Acanthamoeba castellanii

In previous work from this laboratory, a partially purified protein kinase from the soil amoeba Acanthamoeba castellanii was shown to phosphorylate the heavy chain of the two single-headed Acanthamoeba myosin isoenzymes, myosin IA and IB, resulting in a 10- to 20-fold increase in their actin-activated Mg2+-ATPase activities (Maruta, H., and Korn, E.D. (1977) J. Biol. Chem. 252, 8329-8332). A myosin I heavy chain kinase has now been purified to near homogeneity from Acanthamoeba by chromatography on DE-52 cellulose, phosphocellulose, and Procion red dye, followed by chromatography on histone-Sepharose. Myosin I heavy chain kinase contains a single polypeptide of 107,000 Da by electrophoretic analysis. Molecular sieve chromatography yields a Stokes radius of 4.1 nm, consistent with a molecular weight of 107,000 for a native protein with a frictional ratio of approximately 1.3:1. The kinase catalyzes the incorporation of 0.9 to 1.0 mol of phosphate into the heavy chain of both myosins IA and IB. Phosphoserine has been shown to be the phosphorylated amino acid in myosin IB. The kinase has highest specific activity toward myosin IA and IB, about 3-4 mumol of phosphate incorporated/min/mg (30 degrees C) at concentrations of myosin I that are well below saturating levels. The kinase also phosphorylates histone 2A, isolated smooth muscle light chains, and, to a very small extent, casein, but has no activity toward phosvitin or myosin II, a third Acanthamoeba myosin isoenzyme with a very different structure from myosin IA and IB. Myosin I heavy chain kinase requires Mg2+ but is not dependent on Ca2+, Ca2+/calmodulin, or cAMP for activity. The kinase undergoes an apparent autophosphorylation.     

Focal contact assembly through cytoskeletal polymerization: steady state analysis

For many cell types, initial receptor-mediated attachment to a ligand-coated surface is followed by the formation of focal contacts - strong, specialized, discrete adhesive connections between cell and substrate in which receptors are clustered and simultaneously linked to extracellular ligand and cytoskeletal proteins. Since adhesion affects many aspects of cellular physiology including growth, differentiation, and motility, understanding the biochemical factors which regulate focal contact assembly should enhance our understanding of these phenomena. In this paper, we present a mathematical model to examine how receptor-ligand, receptor-cytoskeleton, and cytoskeleton-cytoskeleton interactions affect the formation of receptor clusters which serve as precursors to mature focal contacts. Receptor clustering is presumed to occur through self-recognition of cytoskeletal elements which induce the polymerization of ligand-receptor-cytoskeleton complexes. Polymerization only occurs when the ligand density is above a critical value and a decrease in the receptor-ligand affinity shifts the critical ligand density to higher values. While cytoskeletal protein expression and receptor-cytoskeleton affinity influence the concentration of monomeric complexes, the formation of polymeric ligand-receptor-cytoskeleton aggregates is most sensitive to changes in the self-association affinity between cytoskeletal proteins. We find that a 100-fold enhancement in the affinity between cytoskeletal elements can produce a substantial increase in the total fraction of adhesion receptors associated with focal contact precursors (from 5% to over 90%). Our results suggest that under physiological conditions, cellular control of focal contact assembly most likely occurs through modulation of specific cytoskeletal proteins to solidify cytoskeleton-cytoskeleton connections within precursor focal contact structures.     

Verteilung der A- und B-Zellen sowie des Inselquotienten nach contrainsulärer,antithyreoidaler und antadenohypophysärer Behandlung im Inselorgan der weissen Ratte

Zusammenfassung In einem über 61 Tage laufenden Großversuch wurden 168 erwachsene Ratten in 8 Gruppen geteilt, deren jede anfänglich 21 Tiere enthielt. Im Verlaufe des Versuches verendeten 13 Tiere vorzeitig aus verschiedenen Gründen. Die überlebenden Tiere wurden ausgewertet. Es zeigt sich, daß nach contrainsulärer Behandlung mit Alloxan die B-Zellen des Inselapparates schwer geschädigt, zu einem großen Teil vernichtet sind. Demgegenüber vermehren sich die A-Zellen relativ und absolut, was ein Absinken des nichtnormalisierten Inselquotienten auf ¯Q=0,7983 zur Folge hat. Die alloxandiabetischen Tiere zeigen gegenüber anderen eine signifikant erhöhte Letalität. Schützt man alloxandiabetische Tiere durch chronische antithyreoidale Behandlung mit Methylthiouracil, so kommt es durch Verhinderung der diabetischen Nebennierenhyperplasie und durch Beschränkung der abundanten A-Zellvermehrung zu einer deutlichen Erhöhung der Lebenserwartung.Insgesamt waren bei 155 Ratten N _total=7971 Inseln mit einem Gesamtzellbestand von 319444 A-Zellen und 414 811 B-Zellen ausgezählt worden. Dieses Material war in den 8 Gruppen nichtnormal verteilt, so daß vorerst nur die Verteilungspolygone der A- bzw. B-Zellen und der Inselquotienten nach X ² gruppenweise miteinander verglichen werden konnten. Dieses parameterfreie Verfahren gestattet immerhin schon jetzt die Aussage, daß kombinierte antithyreoidal-antadenohypophysäre Behandlung mit Methylthiouracil + p-Hydroxypropiophenon die mittlere A-Zellzahl/Insel erhöht und somit den Inselquotienten absinken läßt. Daraus kann auf eine Vermaschung der Regelkreise für Inselorgan und Schilddrüse geschlossen und eine Abhängigkeit zumindest der insulären A-Zellen vom hypophysär kontrollierten Endocrinium vermutet werden. Weitere, nach dem gewählten Testverfahren noch nicht statistisch gesicherte Befunde verstärken diesen Eindruck. Eine Vermaschung des B-Zellregelkreises mit dem der Nebennierenrinde kann als gesichert gelten.

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Summary In male adult rats treated with alloxan, in which the B-cells of the pancreatic islets are largely destroyed, the A-cells increase in number, both relatively and absolutely. The high lethality in animals with alloxan diabetes can be reduced by chronic antithyroid treatment with methylthiouracil. This protection is based on (a) the prevention of the adrenal hyperplasia characteristic of the diabetic condition, and (b) a reduction in A-cell proliferation.A large material (7971 islets) was statistically evaluated with the result that the average number of A-cells per islet is increased after combined antithyroid-antadenohypophysis treatment with methylthiouracil and p-hydroxypropiophenon. This indicates a functional relationship between islet tissue and thyroid, more specifically a dependency of at least the A-cells on the endocrine system that is controlled by the adenohypophysis. A functional interdependence between B-cells and adrenal cortex seems to be a certainty.

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Herrn Professor Dr. med. W. Bargmann, Direktor des Anatomischen Institutes Kiel, mit herzlichstem Glückwunsch zum 60. Geburtstag gewidmet.

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Mit dankenswerter Unterstützung durch einen Forschungsauftrag des Staatssekretariates für Hochschulwesen der DDR.     

Marking otoliths of Baltic cod (Gadus morhua Linnaeus, 1758) with tetracycline and strontium chloride

Identified were suitable dosages of tetracycline hydrochloride (TET) in three single treatments and three combined treatments of TET with 2 mg/kg strontium chloride (STR) in wild western Baltic cod (Gadus morhua), in terms of (a) obtainable mark qualities (visibility of fluorescent bands), (b) growth assessment, and (c) induced mortality rates. Isotonic NaCl solution was injected in a control group (25 cod per treatment). The results provide the basis for imperative age validation studies of Baltic Sea cod. Cod originating from pound nets near Fehmarn Island were kept in swimming net cages at the harbor of Warnemünde for 1.5 months. Mean initial total length was 28(±3) cm (salinity: 13, water temperature: 13 to 8°C). Overall average growth of surviving cod was 0.8 mm/day. In single TET treatments, lowest mortality rates and best mark quality were observed for TET concentrations of 100 compared to 50 and 25 mg/kg wet mass. Mortality rates of the 100 mg/kg treatment group were remarkably lower than in the control group emphasizing the antibiotic effect of TET. By contrast, the double treatment in the TET‐STR groups resulted in a binding interaction between both markers in the fish body causing either the antibiotic potency being inhibited or TET and STR forming a non‐beneficial chelate (increased mortality), and decreased incorporation of TET in the otolith (reduced visibility of TET bands). Consequently, TET (short‐term marker) and STR (long‐term marker) should not be injected together. Our results demonstrate that the binding interactions between these substances known from homoiotherm animals also apply for poikilotherms such as fish.     

Nonstereogenic alpha-aminoisobutyryl-glycyl dipeptidyl unit nucleates type I' beta-turn in linear peptides in aqueous solution

The use of alpha,alpha-disubstituted amino acids represents a valuable strategy to exercise conformational control in peptides. Incorporation of the nonstereogenic alpha-aminoisobutyryl-glycyl (Aib-Gly) dipeptidyl sequence into i+1 and i+2 positions of an acyclic peptide sequence, originally designed and investigated by Gellman and coworkers, [H-Arg-Tyr-Val-Glu-Val-Yyy-Xxx-Orn-Lys-Ile-Leu-Gln-NH2] nucleates a stable [2:4] left-handed type I' beta-turn in water. NMR spectra show that this newly designed beta-hairpin does not aggregate in water up to a concentration of approximately 1 mM, and that its backbone conformation is superimposable on corresponding hairpins containing the DPro-Gly (literature) and Aib-DAla (this work) sequences. The Aib-Gly turn-inducer sequence eliminates complications because of cis-trans isomerization of Zzz-Pro bonds, and constitutes an attractive alternative to the proteogenic Asn-Gly and nonproteogenic DPro-Gly motifs previously suggested as turn-inducer sequences. These design principles could be exploited to prepare water-soluble beta-hairpin peptides with robust structures and novel function.