Nove virusne bolesti svinja - izazov za mikrobiologe ili za tehnologe

By investigating the "broiler" type of swine rearing, the paper indicates the possibility for the appearance of new diseases of swine which are unique from the etiological aspect. Namely, these are diseases which probably would not have appeared had production remained at the level which existed during the first half of the twentieth century. Defining of the genetic map of swine, formation of global companies for rearing this animal, requirements of animal proteins for the world population, and the evolution of already existing microorganisms under the pressure of ecological and epizootiological factors, resulted in the formation of an arena in which an attempt must be made to at least maintain a balance between a multitude of negative influences, causative agents of diseases on one side, and production characteristics on the other.Razmatrajući nove bolesti svinja koje idu rame uz rame sa novim tehnologijama uzgoja, potrebno je naglasiti da je broj jedinki ove vrste, tokom istorije pratio broj ljudi. Na primer, u najmnogoljudnijoj zemlji sveta, Kini danas se nalazi oko 500 miliona svinja, slede USA sa 61 milion, Brazil sa 31 i Nemačka sa 25 miliona. To je jedan od razloga što se danas smatra da se savremeni uzgoj ove vrste životinja, pratkično ne razlikuje od brojlerskog načina uzgoja peradi. Kod živine je takav način već tradicionalan i mnogi problemi su već rešeni. Međutim, kod svinja, obzirom na anatomske fiziološke a moglo bi se reći i socijalne karakteristike vrste, "brojlerski" način tova tek je u povoju. Oboljenja kao što su klasična kuga svinja Morbus Aujeszky, crveni vetar, slinavka i šap još se uvek javljaju u pojedinim regionima sveta. Međutim, novi metodi u genetici, tehnologiji držanja i ishrane i strategije kontrole oboljenja, ne mogu priuštiti sebi takav luksuz koji će na primer, na celoj farmi ili čak regionu u državi izvršiti depopulaciju svih jedinki zbog pojave neke od navedenih oboljenja

Grip - influenca

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. .Bilo da se radi o ljudima ili životinjama kao pacijentima, u infektologiji postoje oboljenja koja mogu značajno da smanje populaciju prijemčivih vrsta. Svakako da je pažnja infektologa u proteklih nekoliko decenija usmerena ka onim oboljenjima koja mogu svojom pojavom da 'napune' prve strane tabloida. Treba se setiti spongioformnih encefalopatija, SARS-a ili SIDE. Međutim, istorijski gledano jedno od najopasnijih oboljenja je stari dobri poznanik grip ili influenca. U protekloj deceniji se govorilo o ptičjem i svinjskom gripu. Međutim, radi se o jednom oboljenju svojstvenom većem broju vrsta, koje svoje karakteristike duguje, sa jedne strane, biologiji virusa, a sa druge, epidemiološko-epizootiološkim karakteristikama prijemčivih vrsta. Izmene antigenih struktura koje se dešavaju zahvaljujući mehanizmima reasortiranja segmenata gena uslovljavaju pojavu novih kombinacija tipičnih antigenih molekula virusa (hemaglutinina i neuraminidaze). Kao rezultat, ove izmene mogu da daju one kombinacije H i N molekula koji se po prvi put javljaju u prirodi što znači da nijedna osoba niti jedinka prijemčivih vrsta nije imuna. Samim tim, virusi sa ovakvim kombinacijama antigene strukture, mogu da uslove pandemiju i/ili panzootiju neviđenih razmera, pri čemu treba imati na umu i da u pojedinim populacijama, kako ljudi tako i životinja, mortalitet može da bude veoma visok. Ono što je sigurno jeste da su ptice i svinje rezervoari virusa, ali i žive epruvete u kojima se virus menja, prilagođava vrsti-domaćinu i izlazi iz kruga date vrste-domaćina ka ljudima ili drugoj nekoj vrsti. To znači da kod gripa ne postoji tipično oboljenje za neku od vrsta (ptičji i svinjski) već se radi o infektivnoj bolesti koja je karakteristična za nekoliko vrsta i koja ima potencijal da utiče ne samo na brojnost populacije ljudi i životinja već i na socio-ekonomske prilike u regionima sveta.

Brucella canis at the territory of Serbia in the period from 2004. to 2011.

This study includes examination of dogs from the territory of Serbia during the period from 2004. To 2011. Most of the dogs were from the territory of Belgrade. The total of 193 blood serum samples of proprietary dogs and 120 blood serum samples of stray dogs were tested for the presence of antibodies against Brucella canis. For diagnostics there was used the method of slow serum agglutination in test tubes with 2- mercaptoethanol. During eight years’ period of investigation, out of 193 tested serums taken from proprietary dogs, 29 serum samples, or 15.03%, had undoubtedly positive titre of 1/200. During 2011. Out of 120 tested blood samples taken from stray dogs from Belgrade territory, positive titre of 1/200 had 8 samples, or 6.67%. The results of this investigation point out to a very high seroprevalence of antibodies against B.canis in dogs population from the teritiry of the Republic of Serbia. [Projekat Ministarstva nauke Republike Srbije, br. TR31088

Evidence of Aujeszky s disease in wild boar in Serbia

Background: Aujeszkys disease is a viral disease of suids caused by Suid Herpesvirus 1. The disease has worldwide distribution with significant economic impact. In Serbia, there is neither an Aujeszkys disease eradication nor national vaccination programme of domestic pigs. Since clinical symptoms of Aujeszkys disease are not specific, it is important to establish a link between clinical signs and presence of ADV active infection in wild boars. The aim of this study was to investigate the possibility of active infection within wild boar showing signs of ADV and also to examine relationship between isolates from domestic pigs and wild boar. Having in mind that virus has not been previously isolated from wild boars in Serbia, we report the first isolation of Suid Herpesvirus 1 from this species in Serbia. Results: Tissue and serum samples from 40 wild boars from eastern Serbia were examined for evidence of Aujeszkys disease (AD). Suid Herpesvirus 1 (SHV1), the cause of AD was isolated on PK15 cell line from three tissue samples, inducing cytopathic effect (CPE) with syncytia forming, and viral genome was detected by polymerase chain reaction (PCR) in eight samples. Genetic analysis of us4, us9 and ul49.5 partial sequences showed high homology between ADV isolates from wild boars and between isolates from wild boars and domestic animals. Neutralizing antibodies were not detected by virus neutralisation test (VNT) in sera from four out of eight PCR positive wild boars suggesting recent infection in those animals. Conclusions: This is the first demonstration of Aujeszkys disease virus (ADV) in the wild boar population in Serbia although seroconversion has been detected previously

Ispitivanje pasivnog imuniteta kod prasadi poreklom od krmača vakcinisanih Kina sojem virusa klasične kuge svinja

An experimental study was conducted to investigate the course of classical swine fever (CSF) infection in piglets originating from sows vaccinated with China strain vaccine. The experiment was carried out on 24 piglets (age 28, 35, 44 and 54 days) from vaccinated sows and on 11 non vaccinated piglets, originated from non CSF vaccinated sows. Two piglets from the each age group originating from vaccinated sows were challenged by intramuscular injection with CSF virus. Four piglets of the same age from vaccinated sows, and two piglets derived from unvaccinated sows were added to the challenged group to determine contact (horizontal) infection. After challenge, clinical examination and blood sampling from every animal was carried out on day 0, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29. Blood samples were examined for CSF virus specific antibodies by ELISA test, and for viral antigen i. e. viral RNA by RT-PCR technique. After death or sacrifice pathomorphological changes, presence and distribution of CSF virus antigen were detected in piglet tissue samples by ELISA test. On the basis of the obtained results it can be concluded that not all piglets born to vaccinated sows have maternal antibodies at a detectable level, and the issue of the efficiency of passive immunization needs to be evaluated in the future.Ova istraživanja su izvršena u cilju praćenja toka oboljenja nakon infekcije prasadi virusom klasične kuge svinja (KKS) poreklom od krmača vakcinisanih Kina sojem virusa KKS. Ogled je obavljen na 24 praseta (uzrasta 28, 35, 44 i 54 dana) poreklom od vakcinisanih krmača i na 11 nevakcinisane prasadi poreklom od nevakcinisanih krmača. Po dva praseta iz svake starosne grupe, poreklom od vakcinisanih krmača, su veštački inficirana intramuskularnom aplikacijom virusa KKS. U cilju ustanovljavanja kontaktne (horizontalne) infekcije, direktno inficiranim jedinkama su dodata po četiri praseta istog uzrasta poreklom od vakcinisanih krmača i dva praseta poreklom od nevakcinisanih krmača. Nakon veštačke infekcije, vršen je klinički pregled i uzorkovanje krvi od svake životinje 0, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 i 29 dana ogleda. Uzorci krvi su ispitivani ELISA testom na prisustvo specifičnih antitela protiv virusa KKS, na prisutvo antigena virusa KKS odnosno virusne RNA metodom RT-PCR. Nakon uginuća ili žrtvovanja, vršen je patomorfološki pregled i utvrđivanje prisustva i distribucije antigena virusa u uzorcima tkiva ELISA testom. Na osnovu postignutih rezultata istraživanja može se zaključiti da pojedina prasad, poreklom od vakcinisanih krmača, nemaju detektabilna specifična antitela što nameće razmatranje pitanja efikasnosti pasivne imunizacije

Detection and Phylogenetic Analysis of B2L Gene of ORF Virus from Clinical Cases of Sheep in Serbia

Infection of sheep by the ORF virus (ORFV) is very common in Serbia. ORFV is an economically important viral disease, distributed worldwide. Phylogenetic analysis based on the B2L gene of Serbian ORFV strains from two outbreaks that occurred in Serbia in 2016 is presented in this paper. Crust formation around the lips, nostrils, and udder was noted in all animals from the first outbreak, whilst in the second outbreak, all animals showed swollen and cyanotic lips and muzzle, with no visible crusts. Virus isolation was conducted using Vero cells. Cytopathic effects were evident on the third passage. However, all examined samples were positive using PCR. Phylogenetic analysis of the partial gene sequences (terminal gene regions were not included) encoding B2L gene of Serbian ORFV isolates showed 97.33-100.00% nucleotide and 92.86-100.00% amino acid similarity between each other. However, the viruses were divided into two clusters within the previously recognized Group 2, together with viruses from Croatia, Greece, Finland, China, South Korea and North America. This study is the first report of phylogenetic analysis of ORFV from Serbia and contributes to the data available in the GenBank database. The results of our investigation showed genetic diversity between ORFV strains in Serbia

Suitability of individual and bulk milk samples to investigate the humoral immune response to lumpy skin disease vaccination by ELISA

Background The detection of antibodies against capripoxvirus has become easier with a commercially available ELISA validated for serum and plasma. In order to explore its suitability for immunological investigations on alternative samples, this study targeted milk as sample matrix available through non-invasive sampling. Methods Samples for this study were collected from dairy cows vaccinated against LSD in an area without reported LSD virus circulation. Paired serum and milk (individual and bulk) samples were tested by ELISA without and with modifications of the sample incubation time for the milk samples. For the evaluation of the test specificity, 352 milk samples from a milk repository in Germany were used as negative control. Receiver operating characteristic analysis was performed for determination of the Youden index and determination of the most suitable cut-off value for maximum specificity. Results From 154 analyzed serum samples from Serbia, 75 were detected as positive in the ELISA. Sensitivity and specificity of the ELISA test for milk samples reached values of 88 to 91% using Youden criteria. A cut-off of 10 was determined aiming for maximum specificity. This cut-off value was used for further analysis. Using the protocol for serum, out of 154 milk samples, 38 were detected as positive, number of positive detected milk samples increase up to 48 with modified protocol. Milk samples from Germany reacted negative, except two samples that had borderline results using modified protocol. Significant statistical difference (p < 0.05) was observed between two incubation protocols. The detection of LSD-specific antibodies from bulk milk samples (pools of 2-10 individuals) came along with a reduced sensitivity over the sample of individual animals. Conclusions Results show that the detection of capripoxvirus specific antibodies in milk samples using the commercially available ELISA from IDvet is feasible and can represent a helpful tool for LSDV monitoring programs

Detekcija i genotipizacija izolata virusa klasične kuge svinja u Srbiji

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.Klasična kuga svinja (CSF) je visoko kontagiozno oboljenje svinja koje dovodi do značajnih ekonomskih gubitaka širom sveta. Na osnovu genetske strukture, virus klasicne kuge svinja podeljen je utri genogrupe, od kojih svaka ima tri ili četiri podgrupe. Nedostaju podaci o tome koji genotipovi virusa klasične kuge svinja na teritoriji Republike Srbije cirkulišu u prijemčivoj populaciji. Sekvencioniranjem dela E2 gena i 5’nekodirajućeg regiona (NCR) 15 izolata virusa klasične kuge svinja prikupljenih u periodu od 2006-2008 godine, poreklom od domaćih svinja, dokazano je da svi pripadaju genetskoj grupi 2.3. Uz to, dokazano je i da je upotrebom dva najčešće korišćena real time RT-PCR protokola moguće detektovati sve lokalne izolate virusa klasične kuge svinja koji cirkulišu u Srbiji poslednjih godina, u kojoj se uporedo vršila i intenzivna vakcinacija protiv ove bolesti

Epizootiological service: One of the cornerstones of veterinary medicine

Факултет ветеринарске медицине представља један од факултета који пружају практична знања и вештине, а планови и програми појединих предмета дају солидну основу да по дипломирању лекар ветеринарске медицине зна да постави сумњу на неку заразну и паразитску болест, обави хируршку операцију, анализира резултате дијагностичких испитивања телесних флуида, уведе животињу у анестезију, обави телење, анализира резултате испитивања квалитета сточарских производа, апликује терапију у зависности од дијагнозе и потреба животиња, саветује сточара у вези технологије узгоја и биосиурносних мера итд. Кроз сваки од наведених (и ненаведених) послова, провлачи се један заједнички именитељ. Наиме, појединачни случајеви тј. пацијенти и најчешће позитиван исход третмана, не би значили пуно ако се не би анализовали са становишта правилности и учесталости појављивања болести и успеха спроведених мера. Као лекари ветеринарске медицине, често у помоћ позивамо статистичаре који нам са мање или више успеха, користећи статистичке методе, објасне како се појединачни случајеви болести уклапају у целу слику односа епизоотиолошких детерминанти: узрока (микроорганизма), пријемчиве врсте животиња (макроорганизма) и спољашњег фактора. Међутим, биолошки закони који представљају основу ветеринарске медицине често или нису до краја познати или по природи ствари нису елементи које статистичари уче. Отуда је од великог значаја да сваки лекар ветеринарске медицине буде и епизоотиолог и да на основу својих искустава (рада), покуша да пронађе закономерности поремећаја здравља, производних карактеристика и добробити у популацијама животињских врста као и да установи који су то фактори који утичу на појаву ових поремећаја, а са циљем изналажења мера за њихово уклањање и/или смањењe штета. Имајући у виду професионални профил епизоотиолога, може да се каже да епизоотиолошка служба обједињује податке о поремећајима здравља, производних карактеристика и добробити животиња и има задатак да анализује добијене информације, предлаже и спроводи мере за контролу, сузбијање и искорењивање пре свега заразних болести. Епизоотиолошка служба је у највећем броју случајева (земаља) организована и повезана како по вертикали тако и хоризонтално. Свакако, основа су ветеринари на терену који, поред власника и држаоца животиња, први постављају сумњу на присуство неког поремећаја здравља. Следећи ниво су ветеринарски инспектори и епизоотиолози који се налазе у оквиру специјализованих дијагностичких ветеринарских института. На врху епизоотиолошке пирамиде налази се оделење у оквиру Управе за ветерину које обједињује податке са терена али и тесно сарађује са осталим оделењима Управе. На тај начин, омогућава се да се епизиоотиолошка служба прикупља податке не само о заразним и паразитским болестима које често имају зоонотски потенцијал, већ и информације о поремећајима у продуктивности, подизању квалитативног и квантитативног стандарда производа животињског порекла и ризика по добробит животиња. Исто тако, епизоотиолошка служба у оквиру Управе за ветерину сарађује са Међународним Уредом за епизоотије (ОИЕ), поштујући принципе и критеријуме за размену података и међусобно извештавање Уреда и земаља чланица ОИЕ-а. Епизоотиолошка служба сарађује и са епидемиолошком службом у циљу контроле и сузбијања зооноза.The Faculty of Veterinary Medicine is one of the „hands on“ faculties whose curriculum offers a sound base for a graduate to diagnose an infective or parasitic disease, perform a surgical intervention, analyze results of body fluids tests, assist delivery, assess the quality of livestock products, apply treatment according to the diagnosis and need of the animal, offer council to the farmers, etc. In all the listed (and not listed) duties there is a common denominator, as individual cases would not be of relevance if not observed within the framework of disease regularity and incidence, as well as success of the performed measures. Doctors of veterinary medicine very often appeal to the help of statisticians which to a higher or lesser extent, with the aid of statistical methods, explain how individual cases fit within the whole picture of epizootiological factors: cause (microorganisms), susceptible animal (macro organism), and environmental factors. However, biological rules which represent the core of veterinary medicine very often are not elucidated, or are not taken into account by the statisticians. Thus, the importance for every veterinarian to find the epizootiologist within, and according to his/her knowledge and experience try to define the rules according to which a disease develops or production and welfare are affected. At the same time the veterinarian should be able to define which factors affect the presence of these disorders, all with the aim of prevention and/or damage mitigation. Bearing in mind the professional profile of an epizootiologist, it can be said that the epizootiological service encompasses data on health disorders, productive characteristics and animal welfare and within the Ministry of Agriculture has a role to analyze the obtained data, suggest and carry out control measures, combat and eradicate above all infectious diseases. Also, the Directorate for Veterinary Affairs forwards data to the International Office thus collaborating with the epidemiological service with the aim of zoonoses control and eradication

Simulation of a classical swine fever outbreak in rural areas of the Republic of Serbia

Several different strategies for control and eradication of classical swine fever (CSF) were compared using a Monte Carlo method-based simulation model. The control strategy analysed in this paper, in addition to other CSF control measures, includes application of biosecurity measures on pig farms and rural backyard holdings. Elements of the control strategy are based on applicable regulations and include the simulation of detection of the disease, setting up the protected and surveillance zones, standstill of pig movements and restricted movement of animals, vehicles, equipment, and people with strong control measures in protection and surveillance zones, euthanasia of susceptible pigs, protective vaccination of pigs, compensation etc. During the simulation, different output parameters were compared such as: duration of epidemic of a disease, number of affected holdings and animals, direct costs such as those for dead or culled animals, costs of surveillance, disposal of infectious materials, cleaning and disinfection. Depopulation of affected animals with early diagnostics and vaccination in the protection and surveillance zone proved to be the most effective measures to stop the spread and for the eradication of the disease. Moreover, during the simulation, systematic implementation of biosecurity measures in all pig production clusters was demonstrated to be an appropriate strategy for sustainable control of CSF and the establishment of a stable epidemiological situation