Infestazione con il Dicrocoelium dendriticum e con la Fasciola hepatica sul terreno di caccia dei cervi della speciale riserva naturale Gornje podunavlje (lungo Danubio)

Sažetak Metiljavost (fascioloidoza) je parazitsko oboljenje jetre koje se javlja kod jelena u ritskim ravničarsko šumskim lovištima, rjeđe u planinskim. Uzročnici ove bolesti su dvije vrste metilja, koje pripadaju skupini plosnatih crva, i to veliki (Fasciola hepatica seu Distomum hepaticum) i mali metilj (Dicrocoelium dendriticum). Cilj rada je bio utvrditi utjecaj razvoja bolesti na brojnost jelenske divljači, postupke i mogućnosti liječenja, brojnost jelena nakon primjene lijeka tijekom nekoliko godina, kao i utjecaj ove bolesti na kvalitetu turističke ponude u lovištu specijalnog rezervata prirode "Gornje podunavlje“. Po procjeni lovnih stručnjaka i zdravstvenog stanja odstrijeljenih jelena, invadiranost je stalno rasla, od početnih 20% do konačnih 90%. Metiljavost je nedvosmisleno utjecala na pogoršanje ukupnog zdravstvenog stanja populacije jelena, s vrlo visokim stupnjem smrtnosti. Situacija se počela značajno mijenjati 2006. godine, kada se pristupilo tretmanu jelena s antiparazitskim prepartom (Albendazol) i to putem koncentrirane hrane i soli. Primjena ovakvog tretmana je dala vrlo dobre rezultate.Fluke (fascioliasis) is a parasitic liver disease that occurs in deer in flatland forest and wetland plain hunting areas, rarely in the mou- ntain. The causes of this disease are two types of liver fluke, which belong to a group of flatworms: large fluke (Fasciola hepatica seu Distomum hepaticum) and small (or lancet) fluke (Dicrocoelium dendriticum). The aim of this study was to determine: (a) the impact of disease progression in the number of deer population, methods and treatment options, deer population number after a number of drug treatment continuously over several years, so as the impact of this deer disease at the quality of tourism offer in the area of Spe- cial Nature Reserve “Gornje podunavlje”. According to hunting experts’ estimation, based on established health condition of hunted deer, fluke occupancy has steadily increased, from initial 20% up to 90%. Fluke has, undoubtedly, contributed to the overall health status deterioration of deer population, with very high mortality outcome. The situation has significantly changed in 2006, when the deer ware approached with an anti-parasitic (Albendazole) treatment mixed in a concentrated feed and salt. This treatment showed itself to be very successful.Zusammenfassung Leberegelkrenkheit (Fascioliasis) ist eine Parasitenkrankheit der Leber, die bei den Hirschen im Moorland in ebenen Waldjagdgebie- ten, selten in gebirgigen Gebieten, vorkommt. Die Erreger dieser Krankheit sind zwei Leberegel , die in die Gruppe der flachen Würme gehören, u.zw. der große Leberegel (Fasciola hepatica seu Distomum hepaticum) und der kleine Leberegel (Dicrocoelium dendriti- cum). Das Ziel der Untersuchung war (a), den Einfluss der Krankheitsentwicklung auf die Zahl von Hirschwild festzustellen, Art und Möglichkeiten der ärztlichen Behandlung und die Zahl nach der erfolgten Behandlung mit Medikamenten in Kontinuität durch einige Jahren zu bestimmen. Man sollte auch feststellen, welchen Einfluss diese Erkrankung der Hirsche auf die Qualität des touristischen Angebotes im Jagdgebiet im Naturspezialreservat "Gornje podunavlje“ hat. Nach der Schätzung der Jagdexperten und auf Grund des festgestellten Gesundheitszustand der abgeschossenen Hirsche, wuchs der Grad der Krankheit von anfänglichen 20 % auf 90 %. Die Leberegelkrankheit beeinflusste ohne Zweifel negativ den gesamten Gesundheitszusand der Hirsche, mit einem hohen Grad der Mor- talität. Die Situation änderte sich bedeutend, als die Hirsche 2006 mit dem Antiparasitenpräparat (Albendazol) behandelt wurden, dies durch das konzentrierte Futter und durch Salz. Die Anwendung dieser Behandlung erzielte gute Resultate.Sommario La fasciolosi è una malattia da parassita del fegato che si incontra dai cervi sui terreni di caccia del tipo boschivo in pianura, raramen- te in montagna. La causa di questa malattia sono due tipi della fasciola che appartengono al gruppo di platelminta: sono la fasciola grande (Fasciola hepatica seu Distomum hepaticum) e la fasciola piccola (Dicrocoelium dendriticum). Lo scopo di questa ricerca era determinare l’impatto negativo dello sviluppo di malattia al numero di cervi, ai modi e le possibilità della loro guarigione, e il loro numero dopo l’applicazione continua del rimedio durante alcuni anni, ma si voleva anche studiare l’influsso di questa malattia alla qualità dell’offerta turistica nell’ambito del terreno di caccia dei cervi della speciale riserva naturale Gornje podunavlje. Gli esperti di caccia hanno determinato sui campioni di cervi cacciati un aumento continuo del numero di cervi contagiati, dal 20% iniziale fino al (!) 90% finale. Grazie alla fasciolosi le condizioni salute della popolazione di cervi sono molto peggiorate e hanno avuto una percen- tuale alta di mortalità. La situazione è notevolmente cambiata nel 2006 quando gli professionisti hanno applicato l’antiparassitario Albendazol negli alimenti concentrati e nel sale. I risultati sono stati molto soddisfacenti

<i>Cntn4</i>, a risk gene for neuropsychiatric disorders, modulates hippocampal synaptic plasticity and behavior

Neurodevelopmental and neuropsychiatric disorders, such as autism spectrum disorders (ASD), anorexia nervosa (AN), Alzheimer’s disease (AD), and schizophrenia (SZ), are heterogeneous brain disorders with unknown etiology. Genome wide studies have revealed a wide variety of risk genes for these disorders, indicating a biological link between genetic signaling pathways and brain pathology. A unique risk gene is Contactin 4 (Cntn4), an Ig cell adhesion molecule (IgCAM) gene, which has been associated with several neuropsychiatric disorders including ASD, AN, AD, and SZ. Here, we investigated the Cntn4 gene knockout (KO) mouse model to determine whether memory dysfunction and altered brain plasticity, common neuropsychiatric symptoms, are affected by Cntn4 genetic disruption. For that purpose, we tested if Cntn4 genetic disruption affects CA1 synaptic transmission and the ability to induce LTP in hippocampal slices. Stimulation in CA1 striatum radiatum significantly decreased synaptic potentiation in slices of Cntn4 KO mice. Neuroanatomical analyses showed abnormal dendritic arborization and spines of hippocampal CA1 neurons. Short- and long-term recognition memory, spatial memory, and fear conditioning responses were also assessed. These behavioral studies showed increased contextual fear conditioning in heterozygous and homozygous KO mice, quantified by a gene-dose dependent increase in freezing response. In comparison to wild-type mice, Cntn4-deficient animals froze significantly longer and groomed more, indicative of increased stress responsiveness under these test conditions. Our electrophysiological, neuro-anatomical, and behavioral results in Cntn4 KO mice suggest that Cntn4 has important functions related to fear memory possibly in association with the neuronal morphological and synaptic plasticity changes in hippocampus CA1 neurons

CNTN4 modulates neural elongation through interplay with APP

The neuronal cell adhesion molecule contactin-4 (CNTN4) is genetically associated with autism spectrum disorder (ASD) and other psychiatric disorders. Cntn4-deficient mouse models have previously shown that CNTN4 plays important roles in axon guidance and synaptic plasticity in the hippocampus. However, the pathogenesis and functional role of CNTN4 in the cortex has not yet been investigated. Our study found a reduction in cortical thickness in the motor cortex of Cntn4 -/- mice, but cortical cell migration and differentiation were unaffected. Significant morphological changes were observed in neurons in the M1 region of the motor cortex, indicating that CNTN4 is also involved in the morphology and spine density of neurons in the motor cortex. Furthermore, mass spectrometry analysis identified an interaction partner for CNTN4, confirming an interaction between CNTN4 and amyloid-precursor protein (APP). Knockout human cells for CNTN4 and/or APP revealed a relationship between CNTN4 and APP. This study demonstrates that CNTN4 contributes to cortical development and that binding and interplay with APP controls neural elongation. This is an important finding for understanding the physiological function of APP, a key protein for Alzheimer's disease. The binding between CNTN4 and APP, which is involved in neurodevelopment, is essential for healthy nerve outgrowth

On the Neurobiological Role of the Autism-related Protein Contactin-6

Disruption in the genes coding for the neural cell adhesion molecules contactin-4 (Cntn4), contactin-5 (Cntn5), and contactin-6 (Cntn6) contribute to an increased risk of neuropsychiatric disorders, particularly autism spectrum disorders (ASD). This indicates that the absence or dysfunction of these proteins may disrupt normal brain development. While Cntn4, Cntn5, and Cntn6 are expressed in distinct but overlapping brain regions in rodents, the gross neuroanatomy of the respective null-mutant mice is not affected. More detailed analyses of specific processes in neurodevelopment did demonstrate impairments in several neurobiological processes, including neurite outgrowth, synaptogenesis, neural survival, guiding projections and terminal branches of axons in forming neural circuits. Although studies so far indicate distinct neuroanatomical expression patterns and behavioral phenotypes in the respective null-mutant mice, very little is still known about the molecular mechanisms underlying the biological pathways in which these Cntns participate. The common structural architecture of the Cntn subgroup indicates the necessity of binding partners in order for them to facilitate any neurobiological functions. While Cntns are membrane-linked, but not membrane-spanning, the involved mechanisms of action must rely on a signalling function of the Cntn protein itself or the interaction with other membrane-spanning proteins. Abundant evidence for contactin-1 (Cntn1) and contactin-2 (Cntn2) show that cis- and trans-interaction with several membrane-spanning proteins is part of their repertoire. Therefore, the aim of this thesis is to unravel the neurobiological mechanisms of Cntn4, Cntn5, and Cntn6, with emphasis on Cntn6. Through proteomics novel and known interacting proteins were identified for these three contactins. The interaction of Cntn6 with the cell adhesion G protein-coupled receptor latrophilin-1 (Lphn1, aka CIRL, ADGRL1) was investigated in detail, showing that Cntn6 inhibited Lphn1-mediated apoptosis. These results suggest that Lphn1 functions as a dependence receptor, which induces neuronal apoptosis only in absence of Cntn6. Indeed, neuroanatomical analysis of the visual cortex of Cntn6 null-mutantmice displayed an increase in apoptosis, demonstrating the significant consequences in vivo. Furthermore, it provides a novel look into modes of action of Cntns and into the consequences for brain development. The identification of cell adhesion properties of Cntn6 in the cerebral cortex points to a broader function for Cntn6 in neurodevelopment than was previously shown. This was further emphasized by the ability of Cntn6 to interact with multiple membrane-spanning proteins with each having several interactions of their own, including the well-known and ASD-implicated Nrxn1-Nlgn1 pathway. This may represent the leading edge for integrated protein networks important for brain development, maturation and plasticity. It may provide valuable starting points for future studies to unravel how absence of Cntns impair neurobiological processes, and ultimately lead to ASD pathophysiology

A current view on contactin-4, -5, and -6: Implications in neurodevelopmental disorders.

This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this record.Contactins (Cntns) are a six-member subgroup of the immunoglobulin cell adhesion molecule superfamily (IgCAMs) with pronounced brain expression and function. Recent genetic studies of neuropsychiatric disorders have pinpointed contactin-4 (CNTN4), contactin-5 (CNTN5) and contactin-6 (CNTN6) as candidate genes in neurodevelopmental disorders, particularly in autism spectrum disorders (ASDs), but also in intellectual disability, schizophrenia (SCZ), attention-deficit hyperactivity disorder (ADHD), bipolar disorder (BD), alcohol use disorder (AUD) and anorexia nervosa (AN). This suggests that they have important functions during neurodevelopment. This suggestion is supported by data showing that neurite outgrowth, cell survival and neural circuit formation can be affected by disruption of these genes. Here, we review the current genetic data about their involvement in neuropsychiatric disorders and explore studies on how null mutations affect mouse behavior. Finally, we highlight to role of protein-protein interactions in the potential mechanism of action of Cntn4, -5 and -6 and emphasize that complexes with other membrane proteins may play a role in neuronal developmental functions.Authors of this review were supported by EU-AIMS (European Autism Interventions), which receives support from the Innovative Medicines Initiative Joint Undertaking under Grant agreement no.115300, there sources of which are composed of financial contributions from the European Union's Seventh Framework Programed Grant (P7/2007–2013), from the European Federation of Pharmaceutical Industries and Associations Companies' in-kind contributions, and from Autism Speaks. (K.T.E.K. and J.P.H.B.), by a Fellowship from JSPS (A.O.A. and A.Z)

Tanini pitomog kestena (Castanea sativa)

Extract of chestnut is a mixture of tannic and non-tannic components. The most common tannic components are the glycosides castalagin and vescalagin, which have a therapeutic effect. In conducted studies in vitro and in vivo, it was found that the tannin extract of chestnut has anti-diarrhoeal, astringent, antiviral, antimicrobial, antiprotozoal and anthelmintic properties. At the same time, it was found that the tannin extract of chestnut has anti-carcinogenic properties. Tannin interacts with the membrane structures of certain microorganisms, and thus decreases the permeability of the cell membrane. This antimicrobial activity is advantageous in terms of the action of antibiotics, because tannins only cause the inhibition of microflora growth but not its destruction. Ingested tannins at therapeutic concentrations (2-5%) cannot be absorbed and so cannot cause pharmacodynamic effects on the other organic systems. In the intestines, by means of hydrolysis, they are completely biotransformed into polyphenols (gallic, digallic and elastic acids) and glucose. In the urine, only the degradation products of tannin in the form of glucuronide can be identified. In therapeutic concentrations, tannins have an insignificant effect on the skin, while on mucous membranes they act astringently. At higher concentrations, tannins may lead to degradation of superficial, and even deeper layers of mucous membranes and granulation tissue. Given that they do not show teratogenic, mutagenic or carcinogenic effects, everything indicates that tannins should be more often and more extensively used as safe veterinary drugs

Značenje kliničke primjene timola

Thymol (2-isopropyl-5-methylphenol) is a natural compound from a group of monotherpentinic phenols. These phenols are present in the essential oils of many plants of the Labiateae family, which are used in the folk medicine for centuries. During this long-term period, it is established that thymol has, together with antiseptic and spasmolytic, also antihelmintic action. Thymol antimicrobial spectrum is wide because it acts on Gram-positive and Gramnegative bacteria, fungi and yeasts. As a natural alternative to antibiotics, unlike many traditional antibiotics with bactericidal action, thymol attacks bacterial organelles, especially respiratory chain enzymes, which are linked to mesosomes of the cell membranes, which leads to complete destruction of pathogenic aerobic bacteria results due to a blockage of the cellular respiration and death of the bacteria. During numerous in vitro and in vivo studies conducted with thymol, it has been proved that thymol has antibacterial, antifungal, anti-inflammatory, antiseptic, spasmolytic and anthelmintic effects. It has a wide spectrum of effects against both, Gram-positive and Gram-negative bacteria, yeasts and molds. Nowadays, thymol is used successfully both in human and in veterinary medicine. In veterinary practice, it is used with different species of animals in the treatment of respiratory diseases. Also, it is widely used in beekeeping as a good acaricide at very low concentrations. The exact mechanism of acaridical action of thymol is still not sufficiently known. General idea is that timol acaricidal action is mediated via GABA (A) receptor-R, leading to a neurotoxic effect on the octopaminergic nerves of ticks and insects. Most likely, the death of the ticks is the result of their choking. Thymol is rapidly absorbed from the gastrointestinal system, metabolized by oxidation and glucuronidation, and rapidly (in the first 24 hours of application) excreted via urine, and to a small extent in the unchanged and in large part in the form of glucuronides and /or conjugated sulphates. Additionally, it was found that thymol is quickly reabsorbed from the site of administration, promptly metabolized and rapidly excreted from the organism. Thymol is a natural and safe active substance for which MRL is not established, and it is listed in Annex II of the List of substances without withdrawal period. During the investigation of thymol residues in some bee products (honey and wax) it was found that treatments with this natural substance can be considered to be good alternatives for synthetic acaricides, especially because they do not represent a sanitary risk. Thymol has been evaluated 1992 by the Committee of Expert on Flavorings Substances of the Council of Europe. Thymol is listed among the substances that are permitted as flavouring agents. An upper limit for inclusion of thymol in food has been established at 50 mg/kg and in beverages at 10 mg/kg. During clinical trials of the various medicinal preparations containing thymol on bee-keeping communities, at the established therapeutic concentrations, no adverse effects have been observed. Thymol did not show any theratogenic, mutagenic or carcinogenic effects

Influence of Fasciola Hepatica on Serum Biochemical Parameters and Vascular and Biliary System of Sheep Liver

Background: The aim of this study was to evaluate the functional capacity of the liver based on the activity of specific enzymes and bilirubin in serum and also to investigate the influence of mechanical and toxic effects of Fasciola hepatica on the structures of the blood vessels and biliary tract in the sheep liver.Methods: Blood samples and liver of 63 indigenous sheep of Pramenka breed, slaughtered in the period from March to December 2009 were used. Based on parasitological findings in the liver, all animals were divided into two groups: control (n=34) and infected group (n=29). For investigation and description of pathological changes in sheep liver, naturally infected with F. hepatica, corrosion cast technique was used.Results: Biochemical analysis of tested parameters showed a significant elevation (P≤0.05) of serum gamma-glutamyl transferase (GGT), total bilirubin (TBIL) and direct bilirubin (DBIL) in infected sheep group comparing with the control group. No significant differences were observed for activity of aspartate aminotranferase (AST) between groups. Vascular and biliary systems of the liver were found to be affected.Conclusion: Results of biochemical analysis are consistent with pathological findings and measuring of tested parameters could be used in early diagnosis of sheep fasciolosis and to test the effectiveness of anthelmintic therapy. Corrosion cast technique is very useful for investigation of pathological changes and neoangiogenesis of vascular and biliary system in sheep liver, caused by mechanical and toxic effects of F. hepatica

Contactin-5 expression during development and wiring of the thalamocortical system

The gene encoding the neural cell adhesion molecule Cntn5 (a.k.a. NB-2) has been put forward as a candidate in neurodevelopmental disorders, like autism spectrum disorder (ASD), by recent genetic findings. Little is known about the expression pattern and function of the gene, and its functional involvement in brain development has remained elusive. So far, most research has focused on its early postnatal expression in the auditory system, where the absence of Cntn5 causes abnormal responses to acoustic stimuli and a decrease in fiber density. The current study shows that the Cntn5 gene is expressed in forebrain structures during embryonic development, starting at E15.5, and that it continues to be expressed into adulthood. Sites of strong expression included the thalamus, the caudate putamen (CPu) and to a lesser extent layer Va of the cerebral cortex. Cntn5-positive thalamic nuclei include the laterodorsal (LD), ventrolateral (VL) and posterior group (Po), which contain glutamatergic neurons. Visualization of the expression pattern through the Tau-LacZ fusion protein coded by an insert in the Cntn5 gene, demonstrated that Cntn5-positive nuclei of the thalamus project to the cortex, based on co-localization with thalamocortical markers L1 and Calretinin. These results indicate that the cell adhesion functions of Cntn5 are exploited for circuit formation and connectivity in early development and for synaptic maintenance during adulthood. Subtle alterations in the formation of the thalamocortical circuit may contribute to neurodevelopmental disorders, such as ASD