Comparative examination of the serological response to bluetongue virus in diseased ruminants by competitive and double recognition enzime-linked immunosorbent assays

ΔΕΝ ΔΙΑΤΙΘΕΤΑΙ ΠΕΡΙΛΗΨΗBluetongue (BT) is a viral non-contagious disease of ruminants which is transmitted by insects of the genus Culicoides. In recent years, BT has been a serious threat to livestock and to the economies of European countries. In Serbia the disease appeared for the first time in 2001, and after a 12 year period of freedom, it broke out again in 2014. Considering the actuality of this infectious disease, especially the need for prompt and rapid diagnostics, the aim of this paper was to determine the possibility of detecting the serological response in sheep and cattle with manifested clinical signs of the disease using two different methods: double recognition enzyme-linked immunosorbent assay (sELISA) and competitive enzyme-linked immunosorbent assay (cELISA). A total of 105 blood serum samples of cattle and sheep, which had exhibited clinical signs of BT during 2014, were taken for examination from a serum bank. Out of 74 blood serum samples of sheep and 31 blood serum samples of cattle, 52 samples of sheep and 18 samples of cattle tested positive using sELISA, while 50 samples of sheep and 18 samples of cattle gave positive reactions with cELISA. The results confirm the high sensitivity of sELISA which detected 4% more seropositive sheep in comparison with cELISA. Using Cohen’s kappa statistical analysis, almost perfect agreement was determined between the results (k>0,81) obtained by cELISA and sELISA

Electrical characteristics of female and male human skin

Bioimpedance spectroscopy (BIS) is a popular method for characterizing the electrical properties of biological tissues. In this study, BIS measurement data of female and male human skin were analyzed and compared. The electrical characteristics of tissue were followed according to four-parameters of the Cole-Cole model: low frequency resistance R0; high frequency resistance R∞; relaxation time t and parameter a. Individual electrical characteristics of human skin were determined for 30 women and 30 men. The distribution and one-way analysis of variance (one-way ANOVA) of the Cole-Cole parameters R0, R∞, t, a within the human population indicated their different dependence on gender. Parameter a, which is higher in the female subjects (a =0.83±0.03) than in the male subjects (a=0.7±0.05), is strongly dependent on gender (p=0). Parameter R∞ also significantly depends on gender (p=0.002), while t and R0 seem to be slightly related to gender (p>0.05). [Acknowledgments - This work was supported by the Ministry of Education and Science of the Republic of Serbia (41006).

Detection and genotyping of classical swine fever virus isolates in Serbia

Classical swine fever (CSF) is a highly contagious disease of pigs leading to significant economic losses worldwide. Classical swine fever virus can be classified into three genogroups, each consisting of three or four subgroups. However, there is a lack of knowledge on the genotypes of CSFV isolates in Republic of Serbia. This study, based on the sequences analysis of partial E2 gene and 5' non coding region (NCR) of 15 CSFV isolated during 2006-2008 from domestic pigs, revealed that all were clustered into genetic group 2.3. Additionally, we showed that the two most often used real time RT-PCR assays were able to detect all local CSF viruses circulated in Serbia in the last years during intensive vaccination campaign against CSF. [Projekat Ministarstva nauke Republike Srbije, br. TR 31075 and TR 31088

Monoclonal antibodies as a tool for virus antigen characterization and modification

Od brojnih polja primene (dijagnostika, ispitivanje patogeneze infekcija, terapija tumora), u proteklih nekoliko decenija monoklonska antitela (MAT) su se intenzivno koristila i u karakterisanju virusnih antigena. U zavisnosti od procedure obavljanja pojedinih seroloških reakcija, tačnije - u zavisnosti od načina pripreme antigena, moguće je zaključiti da li se kao antigen koristi cela virusna čestica ili se radi o nekoj njenoj supstrukturi. Koristeći Western blot, radio imuno test i precipitaciju u gelu, kao i neutralizacione testove in vivo (neutralizacija u miševima) i in vitro (plak redukcioni test), ustanovljeno je da se u strukturi virusa uzročnika slinavke i šapa nalaze sekvencionalni (najviše 7) i konformacioni (najviše 12) epitopi. Prilikom obavljanja preliminarnih ispitivanja kompetitivnog radio imuno testa, ustanovljeno je da jedno monoklonsko antitelo, kada formira imuni kompleks sa epitopom za koji je specifično, uslovi pojačavanje vezivanja najmanje tri monoklonska antitela koja su specifična za epitope od kojih je jedan bio sekvencionalan, a dva su bila zavisna od konformacije antigene strukture.Of the many fields of application (diagnosis, testing the pathogenesis of infections, cancer therapy) over the past few decades, monoclonal antibody (mAb) have been extensively used in the characterization of viral antigens. Depending on the procedures for carrying out some serological reactions, and depending on the method of preparing antigen in particular, it is possible to investigate whether the antigen used is a whole virus or was it an its substructure.Using Western blot, radio-immunoassay and gel precipitation and neutralization tests in vivo (neutralization in mice) and in vitro (plaque reduction test), it was found that the in the structure of the virus that causes foot and mouth disease, there are sequential (maximum 7) and conformation (maximum 12) epitopes. During the preliminary investigations in order to prepare competitive radio immunoassay test, it was found that one monoclonal antibody, when in immune complex with the epitope for which it is specific, increased binding at least three monoclonal antibodies specific for epitopes of which one was sekvencionalan and two have been dependent on the conformation of the antigen structure

Antiviral drugs resistance - importance for veterinary medicine

Kada se govori o rezistenciji mikroorganizama, naročito u veterinarskoj medicini, najčešće se misli na sojeve bakterija koji su otporni na hemioterapeutike, prevashodno antibiotike. Uobičajeno dodavanje antibiotika stočnoj hrani, neselektivna, nekontrolisana i često bezrazložna upotreba antibiotika u terapiji i preventivi samo su neki od razloga za ovakav prilaz rezistenciji mikroorganizama. Međutim, u proteklih nekoliko decenija, jedna od najintenzivnijih sfera istraživanja farmaceutske industrije su antivirusni preparati. Može slobodno da se kaže da se o virusima koji izazivaju oboljenja sisara praktično sve zna. Od načina replikacije, mehanizama kojima patogeno deluju na tkiva i organe, ponašanja u inficiranim ćelijama, a za neke viruse je poznat i celokupni genetski kod. Međutim, do danas je za komercijalnu upotrebu registrovano tek tridesetak preparata uz male izglede da se repertoar ovih lekova u dogledno vreme značajnije poveća. Mehanizam delovanja antivirusnih lekova najčešće je inhibicija neke virusne aktivnosti koja je neophodna da se odigra infekcija na nivou ćelije. To su inhibicija adsopcije virusa, fuzije virusa sa ćelijom, uklanjanja virusnih omotača, inhibicija virusnih enzima (virusne DNK polimeraze, reverzne transkriptaze, virusnih proteaza, neuraminidaze, dehidrogenaza i hidrolaza) kao i inhibicija sinteze virusne RNK. Pored potencijalnog značaja nastanka rezistentnih virusa u slučaju da se upotrebe antivirusni preparati kod životinja male (luksuzne) prakse, postoji i opasnost metafilaktične primene antivirusnih lekova kod onih vrsta životinja koje se koriste za ishranu ljudi, za profilaksu virusa koji izazivaju značajne zoonoze.Especially in veterinary medicine, resistance to antimicrobial drugs is most frequently referred to bacterial resistance to chemotherapeutics especially antibiotics that we use in order to cure bacterial diseases in animals. Usual animal feed antibiotic supplementation, nonselective, uncontrolled and often with no reason usage of antibiotic in therapy and prophylaxis are just some of the reasons for such approach to microorganism’s resistance. However, in last few decades development of antiviral drugs is one of the most intensive fields of the work of the pharmaceutical industry. One can easily say that there are a few unanswered questions as far as viruses that cause disease of mammals are concerned. As a science, virology is aware of all aspects of virus biology and infectology. Mechanisms of the virus replication, pathogenic pathways in infected hosts and all aspects of virus biology are well known. Moreover, for most of the important viruses, whole genetic base has been elucidated. In spite of that, up to now there are not more than thirty commercially registered antiviral drugs with a little chance this number to increase. Antiviral repertoire most frequently is inhibition of some virus activity such as virus adsorption, fusion with the cell, virus uncoating, inhibition of virus enzymes (DNA polymerase, reverse transcriptase, neuraminidase, protease, dehydrogenase and hydrolase) as well as virus RNA replication. Apart of the potential importance as a part of the therapy in small-luxury veterinary praxis, antivirus drugs are potential hazard if they are used as a part of metaphylaxis in order to prevent diseases in the animal species that we use as a food. At the same time there is a risk of virus becoming resistant in case of using antiviral drugs in the prophylaxis of the zoonoses

KJU GROZNICA – EPIZOOTIOLOŠKA ENIGMA

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Application of Nature-Based Solutions in Serbian Protected Area Management for the Attainment of Sustainable Development Goals

Healthy natural ecosystems, such as protected areas (PA), support and sustain biodiversity and human well-being by providing essential ecosystem services and health benefits. However, maintaining such ecosystems to provide these environmental benefits becomes a more and more challenging issue with climate changes, biodiversity loss, land degradation and the continued erosion of the natural capital, particularly in the developing world. The main objective of this research is to establish a methodology for achieving the United Nations Sustainable Development Goal 15 “Life on Land”. The methodology will include the determination of heavy metals in soil and needle samples, and questioners for different stakeholders: local people, local businesses, government, civil society organizations (CSO), and the academic community. Soils are integral components of protected area ecosystems. Soil health has been defined as “the capacity of soil to function as a vital living system, within an ecosystem and land-use boundaries, to sustain plant and animal productivity, maintain or enhance water and air quality, and promote plant and animal health” [1]. This definition speaks to the importance of managing soils, so they remain sustainable for future generations. On the other hand, the conifer needles are good bioindicators of ecosystem conditions. The questioner will allow obtaining data about the social and economic benefits of the PAs. The soil and needle samples from four protected areas in Serbia (Zlatibor, Golija, Tara, Đerdap) were analyzed using Inductively coupled plasma –optical emission spectrometry (ICP-OES) and X-Ray Fluorescence (XRF). Quantitative pollution indices were calculated (Enrichment factor (EF), Contamination factor (Cf), Geoaccumulation index (Igeo), Pollution load index (PLI), and Degree of contamination (Cd)), since they are effective tools for converting the raw environmental data into information relevant to support decision-making [2]. Results of this study provide a scientifically-based overview of the conditions of soil health and health of the forest ecosystem and help to propose nature-based solutions (NBS) for enhancing the sustainability of management, especially in the context of improving ecosystem services and climate change adaptation and mitigation. NBS provide affordable, sustainable, and feasible benefits that contribute to improving soil quality and support several ecosystem services relevant to support public health and social well-being [4]. Results of this study use the United Nations Sustainable Development Goals 15 “Life on Land” as a global framework for establishing the partnership between PA managers, conservationists and local authorities that would enhance health and increase environmental, social and economic benefits of the PAs

Influenza - flu

In epidemiology or in epizootiology, there are some infectious diseases that have potential for significant reduction of the susceptible species population. Over the past few decades, epidemiologists were concentrated on diseases that were 'modern' and made front-page news in tabloids. One should recall diseases like bovine spongiform encephalopathy, SARS and AIDS syndromes. However, we should always be aware of the most dangerous diseases such as our old friend, influenza, or simply, flu. In the past decade, we heard about 'bird' or 'swine' influenza. It is the same disease for different animal species as well as for man. Influenza owes its characteristics to specific virus biology as well as to the epidemiology-epizootiology characteristics of the susceptible species. Antigenic changes that took place thanks to reassortment mechanisms of the viral gene segments cause the onset of the new antigenic combinations of the hemaglutinin and neuraminidase molecules. As a result, new H and/or N antigenic formulas appear for the first time in totally susceptible animal and human populations. That means that in such circumstances, no person in the world is immune to the virus. In that case, such a virus can cause a pandemic with disastrous consequences since influenza is a disease with significant mortality, especially in some segments of the human (as well as animal) population. Birds and swine are virus reservoirs, but these species are at the same time live test tubes in which the virus resides, changes and adapts itself not only to the original species but to other species as well. That means that there is no 'bird' or 'swine' flu. Influenza is an infection of several important animal species as well as man that have potential not only for the reduction of the population size but, in case of the human population, for influencing social and economic life.