The detection of parvoviruses and circoviruses in wild boar and jackal populations

Divlje svinje i zlatni šakali su rasprostranjene vrste divljači, a ujedno i rezervoari velikog broja virusa. Infekcija svinja izazvana svinjskim cirkovirusima 2 i 3 (PCV2 i PCV3) je povezana sa multisistemskim sindromom kržljavosti prasadi, pneumonijom, reproduktivnim problemima, sindromom dermatitisa i nefropatije svinja, a virus se može naći i kod asimptomatski inficiranih jedinki. Cirkovirus pasa (CCV) je identifikovan i kao uzročnik vaskulitisa, hemoragične dijareje i granulomatoznog limfadenitisa pasa, a tačna uloga navedenog virusa u infekcijama, kako pasa tako i divljih mesojeda, još uvek predstavlja predmet velikog broja ispitivanja i smatra se da se može prenositi između ovih populacija životinja. Poznato je da svinjski parvovirus 1 (PPV1) izaziva infekciju kod domaćih i divljih svinja koja se karakteriše pobačajima, rađanjem mumificirane prasadi i sterilitetom, a zahvaljujući molekularnim dijagnostičkim metodama otkriveni su i drugi parvovirusi svinja (PPV2-PPV7) sa još uvek neutvrđenim patogenim potencijalom. Parvovirus pasa (CPV) je uzročnik jednog od najvažnijih virusnih oboljenja ove vrste životinja i on, pored gastroenteritisa, dovodi do sistemske infekcije. Uzorci organa, poreklom od odstreljenih divljih svinja i šakala, su ispitivani primenom PCR u skladu sa protokolima uspostavljenim u Laboratoriji za virusologiju, Katedre za mikrobiologiju Fakulteta veterinarske medicine Univerziteta u Beogradu. Ispitani su zbirni uzorci slezine i limfnih čvorova poreklom od 102 divlje svinje pri čemu je detektovano prisustvo PCV2, PCV3, PPV1, PPV2, PPV5 i PPV7 u 18,9, 13,2, 24,5, 73,6, 3,8 i 41,5 procenata uzoraka, tim redom, a utvrđen je i veliki broj mešovitih infekcija. U zbirnim uzorcima slezine i limfnih čvorova, poreklom od 42 šakala, detektovano je prisustvo CCV u 38,1 i CPV u 19 procenata uzoraka. Navedeni rezultati predstavljaju jedan od prvih nalaza PCV3, PPV5, PPV7 i CCV u uzorcima poreklom od divljih životinja u Srbiji i ovo ispitivanje se nadovezuje na naše ranije studije predstavljajući osnovu za dalja planirana istraživanja koja se odnose na genetsku karakterizaciju detektovanih virusa.Wild boars and golden jackals are widespread species and also represent reservoirs of many viruses. Infections of pigs and wild boar caused by porcine circoviruses 2 and 3 (PCV2 and PCV3) are associated with multisystemic wasting syndrome, pneumonia, reproductive problems, dermatitis and nephropathy syndrome, and can also be found in asymptomatic animals. Canine circovirus (CCV) causes vasculitis, hemorrhagic diarrhea, and granulomatous lymphadenitis, however, its exact role in domestic and wild canid infections is still subject to investigation, and it is possibly transmitted between these animal populations. Porcine parvovirus 1 (PPV1) causes infection in domestic and wild pigs, characterized by abortions, stillbirth, and sterility, while other porcine parvoviruses (PPV2-PPV7) are still of unclear pathogenicity. Furthermore, canine parvovirus (CPV) is one of the most important viral pathogens, causing gastroenteritis and systemic infection in canids. Organ samples from wild boars and jackals were examined by PCR using protocols established in the Laboratory of Virology of the Department of Microbiology, Faculty of Veterinary Medicine, University of Belgrade. Pooled samples of spleen and lymph nodes from 102 wild boars were examined, and PCV2, PCV3, PPV1, PPV2, PPV5, and PPV7 were detected in 18.9%, 13.2%, 24.5%, 73.6%, 3.8% and 41.5% of the samples, respectively, with a notable number of mixed infections. The presence of CCV in 38.1% and CPV in 19% of samples were detected in pooled samples of spleen and lymph nodes originating from 42 jackals. These results represent one of the first findings of PCV3, PPV5, PPV7, and CCV in samples from wildlife in Serbia, and this study is sequel of our previous examinations serving as a basis for further research related to the genetic characterization of detected viruses.Zbornik radova i kratkih sadržaj

Genetska analiza i distribucija parvovirusa (ppvs) detektovanih u organima divljih svinja u Srbiji

Porcine parvoviruses (PPVs) are diverse and persistently evolving viruses found in domestic pigs and wild boars. Porcine parvovirus 1 (PPV1) causes reproductive problems in adult animals, although the veterinary relevance of PPV2, PPV3, and PPV4 has not been clarified. The detection and sequence analysis of PPVs circulating in wild boar populations in Serbia was performed to determine their phylogenetic relationships and prevalence in 122 organ samples collected during 2018. The DNA of PPV1, PPV2, and PPV3 was detected in 56.6% of the examined samples, whilst PPV4 was not identified. Overall, PPV3 was the most prevalent in 69.6% of the positive samples, followed by PPV1 in 63.8%, and PPV2 in 21.7% samples. Single infections were more common, although concurrent infections were confirmed in 34.8% samples for two, and 10.1% samples for three viruses. Sequence analysis of wild boar PPV1 showed no significant nucleotide differences from domestic pig PPV1 strains detected in Europe and the USA, however separate clustering from strains from China and the NADL-2 strain was demonstrated. Examination of the selected PPV2 sequences might suggest a certain geographical distribution of genetically diverse PPV2 strains considering high similarities to the strains from neighboring countries, and variability in comparison with other reported PPV2 sequences from different parts of the world. Wild boar PPV3 sequences clustered separately from most of the strains detected in wild boars, as well as the original porcine hokovirus strain. It is further noted that genetically different PPV3 strains circulate amongst Serbian domestic pigs and wild boars.Parvovirusi svinja predstavljaju genetski različite viruse koji izazivaju infekcije doma-ćih i divljih svinja. Parvovirus svinja 1 (PPV1) dovodi do pojave reproduktivnih pro-blema kod odraslih jedinki, dok klinički značaj PPV2, PPV3 i PPV4 još uvek nije u potpunosti razjašnjen. Izvršena je detekcija i analiza genetskih sekvenci parvovirusa koji cirkulišu u populaciji divljih svinja u Srbiji u cilju njihove fi logenetske analize i određivanja zastupljenosti u ukupno 122 uzorka organa prikupljenih tokom 2018. go-dine. Prisustvo DNK PPV1, PPV2 i PPV3 detektovano je u 56,6% ispitanih uzoraka, pri čemu prisustvo PPV4 nije utvrđeno. Među pozitivnim uzorcima, PPV3 je pro-centualno najzastupljeniji virus detektovan u 69,6%, dok je prisustvo PPV1 i PPV2 utvrđeno u 63,8%, odnosno u 21,7% pozitivnih uzoraka. Infekcije jednim virusom su češće identifi kovane, međutim, prisustvo mešovitih infekcija sa dva, odnosno tri par-vovirusa zabeleženo je u 34,8% i 10,1% uzoraka. Analizom genetskih sekvenci PPV1 detektovanih kod divljih svinja nisu utvrđene značajnije razlike u odnosu na analogne sekvence PPV1 poreklom od domaćih svinja iz Evrope i SAD, međutim zabeleženo je izdvajanje u zaseban klaster u odnosu na kineske sojeve virusa i soj NADL-2. Ispi-tivanjem sekvenci PPV2 utvrđena je izvesna geografska distribucija genetski različitih sojeva navedenog virusa s obzirom na njihovu veliku sličnost sa sojevima virusa iz su-sednih zemalja. Sekvence PPV3 detektovanih kod divljih svinja su se na fi logenetskom stablu izdvajale u zaseban klaster u odnosu na većinu dostupnih sekvenci navedenog virusa detektovanih kod divljih svinja. Pored toga, zabeleženo je da genetski različiti sojevi PPV3 cirkulišu u populacijama divljih i domaćih svinja u Srbiji

The First Report of mcr-1-Carrying Escherichia coli Originating from Animals in Serbia

The aim of this study was continuous monitoring of the presence of mcr-1 to mcr-5 genes in Enterobacterales isolated from cattle, pigs, and domestic poultry at intensive breeding facilities in Northern Vojvodina, Serbia, from 1 January 1 to 1 October 2020. Out of 2167 examined samples, mcr-1 was observed in five E. coli isolates originating from healthy turkeys. Four isolates belonged to the phylogenetic group B1, and one isolate to the phylogenetic group A. Detected E. coli serogenotypes (somatic O and flagellar H antigens) were O8:H25 and O29:H25. Core-genome multi-locus sequence typing (cgMLST) revealed three ST58 isolates clustering together in Clonal Complex (CC) 155 and two singletons of ST641-CC86 and ST410-CC23, respectively. Clonotyping revealed CH4-32 (n = 3), CH6-53 (n = 1) and CH4-24 (n = 1). In all isolates, the mcr-1 gene was located on a large IncX4 replicon type plasmid. Eight virulence-associated genes (VAGs) typical of avian pathogenic E. coli (APEC) (fyuA, fimH, hlyF, iss, ompT, sitA, traT, iroN) were detected in four isolates. These isolates were investigated for susceptibility to four biocides and revealed MIC values of 0.125% for glutardialdehyde, of 0.00003–0.00006% for chlorohexidine, of 4–6% for isopropanol and of 0.001–0.002% for benzalkonium chloride. All obtained MIC values of the tested biocides were comparable to the reference strain, with no indication of possible resistance. This is the first report of mcr-1.1-carrying E. coli from Serbia. Although only samples from turkeys were mcr-positive in this study, continuous monitoring of livestock samples is advised to prevent a spill-over from animals to humans

Influence of Carob Flour and Carob Bean Gum on Rheological Properties of Cocoa and Carob Pastry Fillings

The aim of this study was to develop a new cocoa and carob based pastry filling and explore the influences of carob flour and carob gum on the rheological and textural properties, specifically (i) the effect of increasing the amount of carob flour and (ii) the effect of carob bean gum naturally present in the carob flour with seeds versus the commercially available carob bean gum. All samples analyzed in this study exhibited shear thinning behavior. The texture analysis revealed a significant (p < 0.01) increase in consistency and firmness in samples with higher amounts of carob flour added, while higher temperatures significantly (p < 0.01) decreased adhesiveness. When comparing naturally occurring and commercially available LBG (locust bean gum), it was concluded that lower concentrations (up to 0.45% w/w) of naturally occurring LBG work just as well at the same concentrations of commercially available LBG, but this effect cannot be confirmed for higher LBG concentrations, nor for rheological properties determined at higher temperatures (80 °C)

A review of some important viral diseases of wild boars

Wild boars are one of the widest-ranging mammals worldwide and represent reservoirs for many important viruses. Disease outbreaks in domestic swine are often described as a consequence of contact with wild boars, and traditional rearing conditions are a particular risk factor. Examples of such diseases include classical swine fever (CSF), African swine fever (ASF), Aujeszky’s disease (AD), and diseases caused by porcine circoviruses and parvoviruses. Some viral infections causing high mortality rates are easily noticeable and thus reported, though many viruses infecting wildlife are insidious impacting survival rates and reproduction in wild animals. Samples from wild boars for laboratory testing are usually collected postmortem and include various tissues or blood sera. The recovery of viable viruses during virus isolation depends on the virus species and the condition of the sample. Since this method does not yield timely results, most diagnostic procedures are based on PCR or antigen detection methods. Serological surveys are inexpensive and appropriate for prevalence studies. When interpreting the results of diagnostic tests, both virus and host characteristics, and the epizootiological situation must be accounted for. Disease control techniques such as fencing or feeding wild boars cause animal aggregation and give rise to population density which favors pathogen maintenance in the environment. Hunting reduces the number of susceptible animals and is helpful as an additional control measure and for sampling. Available data on infectious disease dynamics in wild boars is scarce, and constant knowledge improvement on pathogenesis, clinical symptoms, risk factors, and adequate control measures are required.Divlje svinje su jedna on najrasprostranjenijih vrsta sisara na planeti, a ujedno predstavljaju i rezervoare mnogih značajnih virusa. Pojava oboljenja u populacijama domaćih svinja se javlja kao posledica kontakta sa divljim svinjama pri čemu tradicionalan način uzgoja životinja predstavlja faktor rizika. Primeri takvih obljenja su: klasična kuga svinja, afrička kuga svinja, Aujeckijeva bolest i oboljenja izazvana svinjskim cirkovirusima i parvovirusima. Određene virusne infekcije sa visokom stopom mortaliteta se mogu lako detektovati, a samim tim i prijaviti, međutim neki virusi divljih svinja ne dovode do vidljivih promena što otežava njihovo otkrivanje. Uzorci za laboratorijska ispitivanja poreklom od divljih svinja se najčešće prikupljaju postmortalno i uključuju različita tkiva ili krvni serum. Uspešnost izolacije virusa u kulturi ćelija zavisi od vrste virusa kao i od stanja dostavljenog uzorka. S obzirom da primena navedene metode oduzima vreme, većina procedura se zasniva na PCR ili metodama detekcije antigena. Pored toga, serološke metode su ekonomski isplative i pogodne za izvođenje studija prevalencije. Prilikom interpretacije rezultata laboratorijskih analiza je izuzetno značajno uzeti u obzir više parametara uključujući osobine virusa i domaćina kao i epizootiološku situaciju na terenu. Metode kontrole zaraznih bolesti divljih svinja poput ograđivanja ili dohranjivanja životinja dovode do povećanja gustine populacije što pogoduje transmisiji patogena. Lov dovodi do smanjenja broja osetljivih životinja u određenoj sredini, međutim koristan je kao dodatna mera kontrole i omogućuje prikupljanje uzoraka. Dostupni podaci o dinamici infektivnih oboljenja divljih svinja su ograničeni i neophodno je konstantno izučavanje njihove patogeneze, kliničkih osobenosti, faktora rizika kao i procena primene određenih mera kontrole

Ispitivanje prisustva adenovirusa pasa u populacijama lisica i šakala

Adenovirusi pasa 1 i 2 (CaDV1 i 2) su genetski i antigenski srodni dvolančani DNK virusi koji ispoljavaju različit tropizam pri čemu se CaDV1 vezuje za receptore ćelija vaskularnog endotela, parenhima jetre i bubrega, dok se CaDV2 umnožava u ćelijama respiratornog, i u određenoj meri crevnog epitela. Infektivni hepatitis pasa je jedna od najznačajnijih zaraznih bolesti ove vrste životinja, a na infekciju CaDV1 osetljivi su i divlji mesojedi dok je oboljenje naročito izraženo kod lisica i protiče sa simptomima encefalitisa. Pseći adenovirus 2 izaziva respiratorno oboljenje i jedan je od uzročnika infektivnog traheobronhitisa pasa. Zlatni šakali (Canis aureus) i lisice (Vulpes vulpes) su široko rasprostranjene vrste divljači, a ujedno i rezervoari velikog broja patogena. Zahvaljujući sveobuhvatnoj vakcinaciji, pojava infektivnog hepatitisa pasa je danas retka i ovo oboljenje se javlja kod nevakcinisanih pasa, odnosno u slučajevima kontakta sa divljim mesojedima. Uzorci poreklom od lisica i šakala odstreljenih na teritoriji AP Vojvodine (Južnog Banata i Zapadne Bačke) analizirani su primenom PCR u skladu sa protokolom za simultanu detekciju CaDV1 i CaDV2 uspostavljenim na Katedri za mikrobiologiju Fakulteta veterinarske medicine Univerziteta u Beogradu. Ispitani su zbirni uzorci organa poreklom od 69 jedinki pri čemu je prisustvo CaDV1 utvrđeno u ukupno 47,8% uzoraka, odnosno kod 34,5% lisica i 57,5% šakala. Za razliku od CaDV1, CaDV2 nije detektovan ni u jednom ispitivanom uzorku. S obzirom da kod odstreljenih životinja nisu utvrđene karakteristične patološke promene, zaključuje se da su inaparentne infekcije izazvane CaDV1 rasprostranjene u populacijama divljih mesojeda u našoj zemlji. Navedeni rezultati predstavljaju najnovije, ako ne i prve podatke o prisustvu psećih adenovirusa kod divljači u Republici Srbiji i predstavljaju osnovu za dalja istraživanja. Uprkos retkoj pojavi infektivnog hepatitisa, vakcinacija pasa ostaje nezaobilazna preventivna mera s obzirom na otpornost CaDV1 u spoljašnjoj sredini, ali i na očiglednu cirkulaciju ovog virusa u populaciji divljih mesojeda.Zbornik radova i kratkih sadržaj

Assessment of Drying Characteristics and Texture in Relation with Micromorphological Traits of Carob (Ceratonia silliqua L.) Pods and Seeds

Carob tree (Ceratonia siliqua L.) is a perennial leguminous evergreen tree native to the coastal regions of the Mediterranean basin and is considered to be an important component of vegetation for economic and environmental reasons. Two constituents of the pod, pulp and seeds, can be used as feed or in food production. In this study, drying characteristics, texture and microstructure of carob pods were studied. Three different carob samples were prepared: whole carob pod, carob pod parts and carob seed. The drying experiments and the modelling showed that carob seeds had the highest drying rate, followed by pod parts and the whole, intact carob fruit. Texture studies showed that the maximum compression force depended on the area of the carob fruit on which compression tests were performed. The seeds showed the highest compression force, followed by the stem zone, the tip and the centre of the fruit. Differences in drying behaviour and texture of carob pods can successfully be interpreted by the micromorphology of the carob pods and seeds. Determining the drying rate, maximum compressive force and micromorphological traits is of great importance for further carob processing (e.g. milling, sieving, carob bean gum production or usage in food or feed products)

Analiza utjecaja mikromorfoloških obilježja plodova i sjemenki na karakteristike sušenja i teksturu rogača (Ceratonia siliqua L.)

Carob tree (Ceratonia siliqua L.) is a perennial leguminous evergreen tree native to the coastal regions of the Mediterranean basin and is considered to be an important component of vegetation for economic and environmental reasons. Two constituents of the pod, pulp and seeds, can be used as feed or in food production. In this study, drying characteristics, texture and microstructure of carob pods were studied. Three different carob samples were prepared: whole carob pod, carob pod parts and carob seed. The drying experiments and the modelling showed that carob seeds had the highest drying rate, followed by pod parts and the whole, intact carob fruit. Texture studies showed that the maximum compression force depended on the area of the carob fruit on which compression tests were performed. The seeds showed the highest compression force, followed by the stem zone, the tip and the centre of the fruit. Differences in drying behaviour and texture of carob pods can successfully be interpreted by the micromorphology of the carob pods and seeds. Determining the drying rate, maximum compressive force and micromorphological traits is of great importance for further carob processing (e.g. milling, sieving, carob bean gum production or usage in food or feed products).Rogač (Ceratonia siliqua L.) je višegodišnja mahunasta zimzelena vrsta rasprostranjena u mediteranskom obalnom području, a iz ekonomskih i ekoloških razloga smatra se važnom komponentom vegetacije tog područja. U proizvodnji prehrambenih proizvoda i stočne hrane koriste se dva dijela mahune rogača: pulpa i sjemenke. U ovom su radu ispitani sljedeći parametri: karakteristike sušenja, tekstura i mikromorfološka obilježja mahuna rogača. Upotrijebljeni su uzorci cijelih mahuna, dijelova mahuna i sjemenki rogača. Rezultati sušenja i aproksimacije eksperimentalno dobivenih podataka matematičkim modelima pokazuju da se sjemenke rogača suše sporije od dijelova mahune ili cijele mahune. Analizom teksture utvrđeno je da je maksimalna sila kompresije ovisila o dijelu mahune na kojem su provedeni kompresijski testovi. Najveću kompresijsku silu imale su sjemenke rogača, zatim peteljka, vrh i središte mahune. Razlike u karakteristikama sušenja i teksturi sjemenki, dijelova mahuna i cijelih mahuna objašnjene su njihovim mikromorfološkim obilježjima. Za daljnju preradu (npr. mljevenje, sušenje, proizvodnju karuba gume ili primjenu u prehrambenim proizvodima i stočnoj hrani) važno je odrediti brzinu sušenja, maksimalnu silu kompresije i mikromorfološka obilježja rogača

The First Report of mcr-1-Carrying Escherichia coli Originating from Animals in Serbia

The aim of this study was continuous monitoring of the presence of mcr-1 to mcr-5 genes in Enterobacterales isolated from cattle, pigs, and domestic poultry at intensive breeding facilities in Northern Vojvodina, Serbia, from 1 January 1 to 1 October 2020. Out of 2167 examined samples, mcr-1 was observed in five E. coli isolates originating from healthy turkeys. Four isolates belonged to the phylogenetic group B1, and one isolate to the phylogenetic group A. Detected E. coli serogenotypes (somatic O and flagellar H antigens) were O8:H25 and O29:H25. Core-genome multi-locus sequence typing (cgMLST) revealed three ST58 isolates clustering together in Clonal Complex (CC) 155 and two singletons of ST641-CC86 and ST410-CC23, respectively. Clonotyping revealed CH4-32 (n = 3), CH6-53 (n = 1) and CH4-24 (n = 1). In all isolates, the mcr-1 gene was located on a large IncX4 replicon type plasmid. Eight virulence-associated genes (VAGs) typical of avian pathogenic E. coli (APEC) (fyuA, fimH, hlyF, iss, ompT, sitA, traT, iroN) were detected in four isolates. These isolates were investigated for susceptibility to four biocides and revealed MIC values of 0.125% for glutardialdehyde, of 0.00003-0.00006% for chlorohexidine, of 4-6% for isopropanol and of 0.001-0.002% for benzalkonium chloride. All obtained MIC values of the tested biocides were comparable to the reference strain, with no indication of possible resistance. This is the first report of mcr-1.1-carrying E. coli from Serbia. Although only samples from turkeys were mcr-positive in this study, continuous monitoring of livestock samples is advised to prevent a spill-over from animals to humans