Procena zdravstvenog rizika povezanog sa konzumiranjem gajenih / divljih dagnji iz Bokokotorskog zaliva

Dagnje su veoma važne za ljudsku ishranu, jer predstavljaju jeftin izvor proteina, koji čine oko 60% suve mase mekog tkiva Mytilus galloprovincialis. Njihovim konzumiranjem omogućen je kako jeftin izvor proteina visoke biološke vrednosti, tako i minerala i vitamina. Sa aspekta hranljivosti, dagnja je važan izvor hrane bogat bitnim elementima (Ca i Fe), kao i određenim vitaminima (B1, B2 i B3). Međutim za dagnje je poznato da poseduju veliki kapacitet akumulacije zagađujućih materija pre svega esecijalnih i neesencijalnih elemenata čiji nedostatak, kao i višak može izazvati ozbiljne zdravstvene probleme kod čoveka. Potrošnja ribe i morskih proizvoda u evropskim zemljama, Francuska (29,7 kg po glavi stanovnika godišnje), Nemačka (12,2), Grčka (22,7), Italija (23,1), Portugal (57,4), Norveška (50), Poljska (9,6), je i dalje niska. Kako ne postoje precizni podaci o prosečnoj potrošnji školjki po glavi stanovnika za većinu zemalja, pa ni za Crnu Goru, dozvoljeni nedeljni unos (PTWI) se koristi za izračunavanje koncentracije elemenata koje čovek može uneti u organizam bez posledica po svoje zdravlje. Dagnja M. galloprovincialis široko je rasprostranjena u priobalnim vodama Bokokotorskog zaliva. Uzgaja se na preko 16 farmi stacioniranih unutar zaliva, ali se duž cele obale zaliva takođe mogu naći i divlje dagnje koje lokalno stanovništvo koristi za ličnu potrošnju. Kako je celokupan zaliv konstantno i sve više izložen negativnom antropogenom uticaju, nameće se potreba za redovnom kontrolom živih organizama iz mora koji se koriste u ishrani čoveka. Cilj ovoga rada bio je da se odredi kvalitet dagnje Mytilus galloprovincialis, odnosno da se proceni eventualni zdravstveni rizik putem konzumiranja ove morske vrste (kultivisane/divlje) sa više lokacija Bokokotorskog zaliva. U svim uzorcima školjki određivane su koncentracije gvožđa, cinka, mangana i bakra. Izmerene koncentracije cinka i bakra u kultivisanim/gajenim dagnjama Bokokotorskog zaliva ispod su vrednosti propisanih regulativom o hrani Crne Gore. Najveće koncentracije cinka i bakra izmerene su u uzorcima divljih dagnji sa lokacije Herceg Novi, dok je najveća koncentracija gvožđa izmerena u kultivisanim dagnjama sa lokacija Kukuljina i Krašići, a najveća koncentracija mangana u kultivisanim uzorcima sa lokacije Bijela. Na osnovu minimalnih i maksimalnih izmerenih koncentracija gvožđa, cinka, mangana i bakra u uzorcima divljih/kultivisanih dagnji i podataka za maksimalno dozvoljeni unos ispitivanih elemenata (PTWI) izračunat je dozvoljeni interval nedeljnog konzumiranja dagnji u kilogramima bez posledica po ljudsko zdravlje za svaki pojedinačno ispitivani element. Nedeljna količina dagnji koju je neophodno konzumirati kako bi se dostigla propisana PTWI vrednost varira: za Fe 11.5 – 14.8 kg i 4.8 – 21.2 kg, za Mn 12.4 – 18.8 kg i 13.4 – 26.4 kg, za Cu 147.0 – 243.0 kg i 198.0 – 233.0 kg, i za Zn 7.1 – 21.7 kg i 14.2 – 17.6 kg za divlje i uzgajene dagnje, respektivno. Na osnovi dobijenih podataka jasno je da se radi o velikim količinama dagnji koje treba konzumirati kako bi se dostigle granične vrednosti PTWIa. Ovo ukazuje da divlje/kultivisane dagnje iz Bokokotorskog zaliva, u odnosu na sadržaj Fe, Zn, Mn i Cu u njima, ne predstavljaju opasnost po zdravlje ljudi i bezbedne su za ljudsku upotrebu

Multiplex PCR for Discriminating Host Plant Associations of Hyalesthes obsoletus (Hemiptera: Cixiidae), a Key Vector and Driver of 'Ca. Phytoplasma solani' Epidemiology

Given the ecological and epidemiological specialization of Hyalesthes obsoletus, the principle vector of ‘Candidatus Phytoplasma solani’, the primary objective of this study was to develop molecular tools for discriminating three host plant associations of the vector populations: (i) Convolvulus arvensis–Urtica dioica (Ca–Ud), (ii) Vitex agnus-castus (Vac), and (iii) Crepis foetida (Cf). The genetic diversity of the nearly full-length mitochondrial COI gene (1467 bp) was analyzed and compared among previously reported and newly collected individuals of the three host plant associations on a wide geographic range. Multiplex PCR was designed and evaluated for discriminating H. obsoletus host plant associations based on the size of amplified fragments: 1084 bp for the Cf association, 645 bp for the Ca–Ud association, and 355 bp for the Vac association. Examples of the epidemiological value of combining data on the genetic characteristics of the vector and the pathogen are provided. The method is intended to facilitate an accurate identification of the vector’s phylogenetic lineage, natural host plant preference, and epidemiological transmission routes of ‘Ca. P. solani’. When applied to H. obsoletus specimens collected from cultivated plants within an agroecosystem and combined with ‘Ca. P. solani’ genotyping, the method should provide valuable information on disease epidemiology, source(s) of emergence, and transmission routes

Attitudes of students from the high medical college of professional studies and nurses towards people suffering from dementia

Dementia is characterized by a progressive decrease in cognitive functions, and the term includes different etiologies. Cognitive decline includes loss of memory and deterioration in executive functions, such as planning and organizing skills, sufficient to influence social activities. The aim of this study was to examine and compare the attitudes (knowledge, emotions and behaviour) of students at the High Medical College of Professional Studies and nurses towards people suffering from dementia. The study was de-signed as a qualitative study with the use of a questionnaire. The Dementia Attitudes Scale (DAS) was used in our study. A total of 283 respondents answered the survey: 56.25% were students, and 43.75% were nurses. The internal consistency of the DAS was found to be good with a Cronbach's α of 0.792. In the overall score for attitudes, a significant difference was found between students (100.47±10.91) and nurses (95.51±16.10). The students had a better score regarding questions describing their behaviour towards these individuals (p<0.001) and emotions for these patients (p<0.001). For knowledge, there was no difference between the two groups of subjects (p=0.901). Regarding the overall score, attitudes of students and nurses towards people with dementia were positive. This research suggested that the training of senior team members who then had dementia expertise was a key component in developing attitudes and improving care practices and outcomes for these patients. Continuous education of all medical staff who have contact with people who suffer from dementia is important

Symptomatology, (Co)occurrence and Differential Diagnostic PCR Identification of ‘Ca. Phytoplasma solani’ and ‘Ca. Phytoplasma convolvuli’ in Field Bindweed

Field bindweed (Convolvulus arvensis) is one of the major natural plant hosts and reservoirs of ‘Candidatus Phytoplasma solani’ (‘Ca. P. solani’), the causal agent of plant diseases in diverse agricultural crops, including Bois noir (BN) disease of grapevine. Phylogenetically, the most closely related phytoplasma to ‘Ca. P. solani’, the ‘Ca. P. convolvuli’, induces disease in field bindweed that is known by its symptoms as bindweed yellows (BY). The occurrence, coinfection and symptoms association of the two phytoplasmas in shared host plants were the subject of this study. Specific primers for the amplification of the elongation factor Tu gene (tuf ) were developed for the identification of ‘Ca. P. convolvuli’ (by conventional nested PCR), as well as primers for simultaneous detection of ‘Ca. P. solani’ and ‘Ca. P. convolvuli’ by duplex SYBR Green real-time PCR. Among symptomatic bindweed plants, 25 and 41% were infected with a single phytoplasma species, ‘Ca. P. solani’ and ‘Ca. P. convolvuli’, respectively, while 34% were infected with both phytoplasmas. None of the non-symptomatic control plants carried phytoplasma, while non-symptomatic plants from our previous epidemiological studies in BN-affected vineyards were confirmed to be infected solely with ‘Ca. P. solani’. Stamp gene typing revealed Rqg50 and Rqg31 ‘Ca. P. solani’ genotypes in plants coinfected with ‘Ca. P. convolvuli’, while three diverse genotypes (Rqg50, GGY and Rpm35) were identified in a single locality with symptomatic bindweeds infected solely with ‘Ca. P. solani’. Variations in symptoms and their association with each of the phytoplasmas are described and documented. The symptom of bushy appearance could be single out as specific for ‘Ca. P. convolvuli’ infection, while occurrence of ‘Ca. P. solani’ could not be unequivocally associated with specific alterations in infected bindweeds. The results are discussed in the context of the epidemiological and ecological complexity of ‘Ca. P. solani’-induced diseases and the relationship between the two phytoplasma relatives in shared host plant

Retrospective use of integrative taxonomy in classical biological control: The unintentional introduction of the weevil Rhinusa dieckmanni to North America

A seed-feeding weevil introduced to North America (NA) as a biological control agent of the invasive toadflax Linaria dalmatica (L.) Mill., identified then as Gymnetron antirrhini “Dalmatian host race” and subsequently confirmed as established, was revealed through our study to be a separate species, i.e., Rhinusa dieckmanni (Behne) (Coleoptera: Curculionidae). This weevil species was presumed to be endemic in its native range, with a distribution restricted to Mount Rila in southwestern Bulgaria. We conducted a comprehensive study of seed-feeding weevils associated with L. dalmatica, L. dalmatica ssp. macedonica (Griseb.) D.A. Sutton, L. genistifolia (L.) Mill., and L. grandiflora Desf. across a broad geographic area of their native range. Those results revealed that all four host plants were used by R. dieckmanni and thus the native geographic range of the species is wider than expected, encompassing the Balkans and the Anatolian Plateau. Our observations suggest that phenotypes of this weevil are highly variable and dependent on the seed capsule size of the Linaria host population. The haplotype network based on mitochondrial COII, 16S genes, and nuclear EF 1-α gene genealogy confirmed the conspecific nature of geographically distant weevil populations, that is, R. dieckmanni phenotypes utilizing L. genistifolia, L. dalmatica, and L. grandiflora for larval development. Specimens collected from L. dalmatica in the northwestern USA shared the same haplotypes as samples from L. dalmatica ssp. macedonica in southwestern North Macedonia, supporting the known introduction history of the North American population. Females from these populations have relatively short rostrums, which may limit their reproductive success on North American invasive L. dalmatica with larger seed capsules

Genetic Diversity of Flavescence Dorée Phytoplasmas in Vineyards of Serbia: From the Widespread Occurrence of Autochthonous Map-M51 to the Emergence of Endemic Map-FD2 (Vectotype II) and New Map-FD3 (Vectotype III) Epidemic Genotypes

Flavescence dorée (FD) is a European quarantine disease of grapevine caused by FD phytoplasma (FDp) transmitted by the leafhopper of North American origin Scaphoideus titanus. The disease affects the most important viticultural regions of Europe and all wine-growing regions of Serbia. Unlike the insect vector, the pathogen is native to Europe and associated with several wild host plants among which alder trees as the main source of two out of three map genetic clusters of pathogen variants (Map-FD1 and FD2). Heretofore, the FDp epidemic in Serbian vineyards was thought to be monotypic, i.e., caused by the single genotype of the Map-FD3 cluster, M51, and correlated with clematis as the natural source plant. This study aimed to provide data on genetic diversity, through map and vmpA gene typing, and insights into ecological properties of epidemiological cycles driving the epidemic outbreaks of FD in Serbia today. Map genotyping of 270 grapevine isolates collected from 2017 to 2019 confirmed M51 as autochthonous genotype widespread in all wine producing regions of Serbia and the dominant FDp epidemic genotype in most of the districts (75%, 202/270 isolates), except in north Serbia where multiple outbreaks of M12 Map-FD3 were recorded (54 isolates). Tree of heaven is reported as a new FDp plant reservoir for the Serbian vineyards, hosting the M51 genotype, along with clematis. An outbreak of a new endemic Map-FD3 genotype M144 was documented in grapevine samples from east Serbia (5 isolates), correlating with previous findings of the same genotype in clematis. In addition, single grapevine infections with five new Map-FD3 genotypes (M150-M154) were recorded in central Serbia, thus indicating high endemic potential for new outbreaks. The vmpA typing placed all Map-FD3 isolates into the VmpA-III cluster, i.e., Vectotype III. Finally, we found direct evidence that at least two FDp endemic genotypes, M89 and M148, of the Map-FD2/VmpA-II have escaped from alders and propagated in the grapevine-S. titanus pathosystem in Serbia (Vectotype II). Our findings confirm the high complexity of the FDp ecological cycle and provide evidence of a unique, autochthonous Balkan epidemiology sourced endemically

Plasmopara obducens – nova pretnja proizvodnji Impatiens walleriana u Srbiji

During 2010, Impatiens walleriana plants with symptoms of downy mildew were collected in a greenhouse in the vicinity of Mionica, Kolubara District. Disease incidence was extremely high, approaching 100%, and wilting and collapse of affected plants was very rapid, resulting in losses of more than 90%. White downy growth produced on the lower leaf surface consisted of hyaline, thin–walled sporangiophores with monopodial branching and numerous, ovoid and hyaline sporangia. Apical branchlets of sporangiophores were at right angles to the main axis, with no apical thickening. Pathogenicity tests included inoculation of young I. walleriana plants by spraying with a sporangial suspension, and downy mildew symptoms were observed after 13 to 15 days. The absence of well–defined spots on the infected impatiens leaves and straight sporangiophores indicated that the pathogen was P. obducens, which was further supported by molecular identification, the 5’–end of the nuclear DNA coding for the large ribosomal subunit (LSU rDNA) was amplified by PCR, using primers NL1 and NL4. A representative isolate, 28–10, was sequenced and phylogenetic analysis showed its grouping with other P. obducens isolates of different origin. Considering that impatiens downy mildew in Serbia is proved to be caused by P. obducens it is necessary to employ adequate phytosanitary measures to prevent further spread of the pathogen.Tokom 2010. godine biljke Impatiens walleriana sa simptomima plamenjače sakupljene su u stakleniku u okolini Mionice, Kolubarski okrug. Učestalost oboljenja bila je izuzetno visoka, blizu 100%, a sušenje i propadanje zaraženih biljaka veoma brzo, što je dovelo do šteta većih od 90%. Bela prevlaka patogena koja se razvijala na naličju listova sastojala se od hijalinskih monopodijalno razgranatih sporangiofora sa tankim zidovima i brojnih, ovoidnih i hijalinskih sporangija. Vrhovi grana sporangiofora granali su se pod pravim uglom u odnosu na glavnu osu, bez vršnog zadebljanja. Testovi patogenosti uključili su inokulacije mladih biljaka I. walleriana prskanjem suspenzijom sporangija, a simptomi plamenjače razvili su se nakon 13-15 dana. Odsustvo definisanih pega na zaraženim listovima impatiensa i prisustvo pravih sporangiofora ukazalo je da je prouzrokovač P. obducens, što je dalje potvrđeno molekularnom identifikacijom. Primenom lančane reakcije polimeraze (polymerase chain reaction, PCR), pomoću prajmera NL1 i NL4, umnožen je kodirajući 5’-kraj DNA velike ribozomalne podjedinice (LSU rDNA). Amplifikovani produkt odabranog izolata 28-10 je sekvencioniran, a filogenetske analize pokazale su njegovo grupisanje sa ostalim izolatima P. obducens različitog porekla. Uzimajući u obzir da je ovim istraživanjima dokazano da je prouzrokovač plamenjače na Impatiens walleriana u Srbiji P. obducens, neophodno je preduzimanje odgovarajućih fitosanitarnih mera u cilju sprečavanja daljeg širenja patogena u našoj zemlji

Epidemiological Role of Dictyophara europaea (Hemiptera: Dictyopharidae) in the Transmission of ‘Candidatus Phytoplasma solani’

Bois noir, an economically important disease of grapevine yellows that causes significant economic losses in wine production, is associated with ‘Candidatus Phytoplasma solani’ and transmitted to grapevines by cixiids Hyalesthes obsoletus and Reptalus panzeri. Polyphagous planthopper Dictyophara europaea, commonly found in natural habitats, harbors phytoplasmas from distinct groups and is an alternative vector in the open epidemiological cycles of the Flavescence dorée phytoplasma in grapevine in European vineyards. This study addresses the role of D. europaea in the transmission cycle(s) of ‘Ca. P. solani’ among wild habitats, natural reservoir plants, and the vineyard agroecosystem using MLSA and transmission trials with naturally infected adults to grapevine and Catharanthus roseus. The infection rates of D. europaea ranged from 7% to 13% in diverse locations, while reservoir herbaceous plants were infected in the amount of 29%. A total of 13 CaPsol MLSA genotypes were detected in D. europaea (7) and plants (8). Nine of them corresponded to previously identified genotypes. Two new genotypes were found in D. europaea (tuf-b1/S1/V14/Rqg50-sv1 and tuf-b1/S18/V14/Rqg50-sv1) and one in Convolvulus arvensis (tuf-b1/S1/V2-TA/Rqg31-sv1), whereas one was shared by two hosts, Crepis foetida and Daucus carota (tuf-b1/S1/V2-TA/STOL-sv1). Naturally infected D. europaea successfully transmitted the tuf-b1/S1/V2-TA/STOL type to five grapevines and six periwinkles, tuf-b1/S1/V2-TA/Rqg31 to one grapevine, and tuf-b1/S1/V2-TA/Rqg50 to one periwinkle, indicating that D. europaea is an intermediate vector in CaPsol epidemiological cycles

The “code red” for Balkan vineyards: occurrence of Orientus ishidae (Matsumura, 1902) (Hemiptera: Cicadellidae) in Serbia

Orientus ishidae (Matsumura, 1902) (Hemiptera: Cicadellidae), known as the mosaic leafhopper, is an invasive alien species native to the Eastern Palaearctic, but also widespread and well established on the eastern territory of North America. Since its first detection in Europe in 1998, this polyphagous species has expanded its distribution area and rapidly spread through central and western European countries, inhabiting a wide range of broadleaf trees and shrubs (e.g. Gleditsia triacanthos, Salix spp., Corylus spp., Acer spp., Betula spp., Populus spp. and Carpinus spp.), which are usually present in the surrounding of vineyards. Over the last decade, O. ishidae was frequently found within European vineyards and was proven to be able to complete its life cycle on grapevine. Moreover, this leafhopper was shown to be naturally infected with the Flavescence dorée phytoplasma (FDp), a devastating disease which causes significant economic losses in the major vine-producing countries. This study provides data on the occurrence of O. ishidae in Serbia, clearly confirming that it has expanded its distribution range to the Balkan Peninsula which could lead to establishment of new FDp epidemiological cycles inside the local vineyard regions. As a consequence, there could be an increased negative impact on phytosanitary situation with a continual spread of this vector species and FDp epidemic outbreaks in Southeastern Europe

Identification of phenolic compounds in cecidogen and Linaria vulgaris. Changes in phenolic metabolism during gall formation induced by Rhinusa pilosa

Rhinusa pilosa (Gyllenhaal) (Coleoptera, Curculionidae) induces galls in Linaria vulgaris Mill., Yellow or common toadflax (Plantaginaceae). Gall is defined as atypical plant growth induced by host-specific organisms, and represents larval chamber which provides food and protection during insect development. It has been shown that gall formation is triggered by ovoipositional fluid (cecidogen). In order to determine which cecidogen compounds may have bioactive properties, we performed comparative analysis of phenolics profiles of cecidogen and stem extracts of L. vulgaris using UHPLC coupled with OrbiTrap mass analyzer. In addition, we analyzed the effects of feeding, oviposition and early gall development on Class III peroxidase activity (POD, EC 1.11.1.7) and phenolic content in the stems of L. vulgaris during seven days. Out of 55 phenolic compounds identified in cecidogen and L. vulgaris stem, one unknown phenolic glycoside was found only in cecidogen, which produced an MS2 base peak at 387 m/z, and 327 m/z and 267 m/z base peaks at MS3 and MS4 fragmentation, respectively. Gall development during seven days was accompanied by decreased concentrations of hydroxybenzoic, hydroxycinnamic acids and flavonoids and by slight lignin deposition. An oscillatory induction of POD activity, with the first peak obtained 3 h after oviposition, was related to oxidative burst during stem wounding and oviposition. Overall results suggested the importance of phenolics in stem structural changes and regulation of plant metabolism induced by R. pilosa