Ecology and significance of the red fox (Vulpes vulpes L.) and jackal (Canis aureus L.) as a natural reservoir of multilocular echinococcosis and other intestinal zoonosis in Vojvodina

U periodu od 2015. do 2018. godine ispitane su 223 lisice i 64 šakala sa područja Vojvodine Parazitološkim pregledom konstatovano je prisustvo 9 vrsta crevnih helminata, od toga jedna vrsta metilja (Alaria  alata), četiri vrste pantljičara (Mesocestoides spp., Ecinococcus multilocularis, Taenia pisiformis, Taenia hydatigena) i četiri vrste nematoda (Toxocara canis, Toxascaris leonina, Uncinaria stenocephala, Pterygodermatites affinis). Nematoda Pterygodermatites affinis je prvi put zabeležena u Vojvodini. Izolovane su 4483 jedinke parazita, od toga 3826 kod lisica i 657 kod šakala. Zabeležena je veća zaraženost lisica (79.8%) u odnosu na šakale (57.8%) (p=0.0003). Najveći procenat obe vrste domaćina (Vulpes vulpes -  49.3%;  Canis aureus - 42.2%) bio je zaražen pantljičarama roda Mesocestoides. Kod obe vrste domaćina, vrednosti Berger-Parkerovog indeksa ukazale su na relativno ujednačenu respodelu parazita po vrstama u ukupnom uzorku, dok su sva tri indeksa (Šenonov, Simpsonov i Berger-Parkerov) pokazala su da je područje Bačke sa najvišim biodiverzitetom zajednica helminata. Za vrste Echinococcus multilocularis i Alaria alata utvrđena je statistički značajna razlika u procentu inficiranih lisica u zavisnosti od područja Vojvodine. Prema taksonomskoj strukturi helmintofaune kod lisica je dokazana najveća procentualna zastupljenost mešovite infekcije (61%), dok se broj vrsta po domaćinu kretao od 1 do 5. Registrovane su 42 različite kombinacije vrsta parazita. Kod šakala se takođe najčešće javljala mešovita infekcija (59%), dok se broj vrsta po domaćinu kretao od 1 do 4. Registrovano je 11 kombinacija crevnih parazita.  Koprološki test se u ovom istraživanju pokazao kao nisko osetljiv i manje specifičan za identifikaciju parazita u odnosu  na  direktan  pregled  intestinuma  nakon  autopsije životinje.  Prevalenca infestiranih lisica (56.3%) i šakala (48%) dokazana koprološkom metodom manja je u odnosu na prevalencu prema patološkom nalazu. Kod obe vrste domaćina nije registrovan statistički značajan uticaj pola na prevalencu infekcije. Od identifikovanih vrsta helminata lisica i šakala, više od polovine njih ima zoonotski ili potencijalni zoonotski značaj: Alaria alata,  Mesocestoides spp., Echinococcus multilocularis, Toxocara  canis i Uncinaria stenocephala. Najvažnija među njima je pantljičara E.  multilocularis, uzročnik ehinokokoze, bolesti svrstane na listu zoonoza od prioriteta na području Srbije. Na području Srema dokazana je pojava žarišta multilokularne ehinokokoze, zbog čega je neophodna optimizacija strategije za kontrolu širenja date vrste parazita.During the period from 2015. to 2018. year, 223 red foxes and 64  jackals  from  the  area  of  Vojvodina  were  examined. Parasitological  examination  revealed  9  species  of  intestinal helminths,  thereof  one  species  of  fluke  (Alaria  alata),  four species  of  tapeworms  (Mesocestoides  spp.,  Ecinococcus multilocularis,  Taeniapisiformis,  Taenia hydatigena)  and four species  of  nematodes  (Toxocara  canis,  Toxascaris  leonina, Uncinaria  stenocephala,  Pterygodermatites  affinis).  The nematode Pterygodermatites affinis was recorded in Vojvodina for the first time. There were isolated 4483 individuals of the parasite, thereof 3826 at red foxes and 657 at jackals. The higher infestation  of  foxes  (79.8%)  compared  to  jackales  (57.8%) (p=0.0003) was recorded. The highest percentage of both hostspecies  (Vulpes  vulpes-  49.3%;  Canis  aureus-  42.2%)  was infected with tapeworms of the genus  Mesocestoides.  In both host species, the value of the Berger-Parker index indicated a relatively uniform  distribution of parasites by species in the total sample, while all three indices (Shannon, Simpson and BergerParker)  showed  that  the  area  of  Bačka  had  the  highest biodiversity  of  helminth  community.  For  the  species Echinococcus multilocularis and  Alaria  alata  was confirmed a statistically significant difference in the percentage of infected foxes  depending  on  the  area  of  Vojvodina.  According  to  the taxonomic structure of helminth fauna, in foxes was proven the highest percentage of mixed infection (61%), while the number of species per host ranged from 1 to 5. There were registered 12 different combinations of parasite species. In jackals, the mixed infection was also the most common  (59%), while the number of  species  per  host  ranged  from  1  to  4.  There  were  also registered  11  combinations  of  intestinal  parasites.  The coprologic test in this study proved to be low sensitiv and less specific  for  identification  of  parasites  compared  to  direct examination  of  the  intestine  after  autopsy  of  the  animal.  The prevalence of infested foxes (56.3%) and jackals (48%) proven by  the  coprological  method  was  lower  compered  to  the prevalence of patological finding. The statistically significant influence  of  sex  on  the  prevalence  of  infection  was  not registered in neither one of the host species. Of the identified species  of  helminths,  at  foxes  and  jackals,  more  than  half  of them  have  zoonotic  or  potential  zoonotic  significance:  Alaria alata,  Mesocestoides  spp.,  Echinococcus  multilocularis, Toxocara canis  i  Uncinaria stenocephala.  The most important among  them  is  the  tapeworm  E.  multilocularis,  the  cause  of echinococcosis, a disease classified as a zoonosis of priorities in Serbia. In the area of Srem, the occurence of foci of multilocular ecinococcosis  was  proved,  which  is  why  it  is  necessary  to optimize the strategy to control the spread of parasites

Ecology and significance of the red fox (Vulpes vulpes L.) and jackal (Canis aureus L.) as a natural reservoir of multilocular echinococcosis and other intestinal zoonosis in Vojvodina

U periodu od 2015. do 2018. godine ispitane su 223 lisice i 64 šakala sa područja Vojvodine Parazitološkim pregledom konstatovano je prisustvo 9 vrsta crevnih helminata, od toga jedna vrsta metilja (Alaria  alata), četiri vrste pantljičara (Mesocestoides spp., Ecinococcus multilocularis, Taenia pisiformis, Taenia hydatigena) i četiri vrste nematoda (Toxocara canis, Toxascaris leonina, Uncinaria stenocephala, Pterygodermatites affinis). Nematoda Pterygodermatites affinis je prvi put zabeležena u Vojvodini. Izolovane su 4483 jedinke parazita, od toga 3826 kod lisica i 657 kod šakala. Zabeležena je veća zaraženost lisica (79.8%) u odnosu na šakale (57.8%) (p=0.0003). Najveći procenat obe vrste domaćina (Vulpes vulpes -  49.3%;  Canis aureus - 42.2%) bio je zaražen pantljičarama roda Mesocestoides. Kod obe vrste domaćina, vrednosti Berger-Parkerovog indeksa ukazale su na relativno ujednačenu respodelu parazita po vrstama u ukupnom uzorku, dok su sva tri indeksa (Šenonov, Simpsonov i Berger-Parkerov) pokazala su da je područje Bačke sa najvišim biodiverzitetom zajednica helminata. Za vrste Echinococcus multilocularis i Alaria alata utvrđena je statistički značajna razlika u procentu inficiranih lisica u zavisnosti od područja Vojvodine. Prema taksonomskoj strukturi helmintofaune kod lisica je dokazana najveća procentualna zastupljenost mešovite infekcije (61%), dok se broj vrsta po domaćinu kretao od 1 do 5. Registrovane su 42 različite kombinacije vrsta parazita. Kod šakala se takođe najčešće javljala mešovita infekcija (59%), dok se broj vrsta po domaćinu kretao od 1 do 4. Registrovano je 11 kombinacija crevnih parazita.  Koprološki test se u ovom istraživanju pokazao kao nisko osetljiv i manje specifičan za identifikaciju parazita u odnosu  na  direktan  pregled  intestinuma  nakon  autopsije životinje.  Prevalenca infestiranih lisica (56.3%) i šakala (48%) dokazana koprološkom metodom manja je u odnosu na prevalencu prema patološkom nalazu. Kod obe vrste domaćina nije registrovan statistički značajan uticaj pola na prevalencu infekcije. Od identifikovanih vrsta helminata lisica i šakala, više od polovine njih ima zoonotski ili potencijalni zoonotski značaj: Alaria alata,  Mesocestoides spp., Echinococcus multilocularis, Toxocara  canis i Uncinaria stenocephala. Najvažnija među njima je pantljičara E.  multilocularis, uzročnik ehinokokoze, bolesti svrstane na listu zoonoza od prioriteta na području Srbije. Na području Srema dokazana je pojava žarišta multilokularne ehinokokoze, zbog čega je neophodna optimizacija strategije za kontrolu širenja date vrste parazita.During the period from 2015. to 2018. year, 223 red foxes and 64  jackals  from  the  area  of  Vojvodina  were  examined. Parasitological  examination  revealed  9  species  of  intestinal helminths,  thereof  one  species  of  fluke  (Alaria  alata),  four species  of  tapeworms  (Mesocestoides  spp.,  Ecinococcus multilocularis,  Taeniapisiformis,  Taenia hydatigena)  and four species  of  nematodes  (Toxocara  canis,  Toxascaris  leonina, Uncinaria  stenocephala,  Pterygodermatites  affinis).  The nematode Pterygodermatites affinis was recorded in Vojvodina for the first time. There were isolated 4483 individuals of the parasite, thereof 3826 at red foxes and 657 at jackals. The higher infestation  of  foxes  (79.8%)  compared  to  jackales  (57.8%) (p=0.0003) was recorded. The highest percentage of both hostspecies  (Vulpes  vulpes-  49.3%;  Canis  aureus-  42.2%)  was infected with tapeworms of the genus  Mesocestoides.  In both host species, the value of the Berger-Parker index indicated a relatively uniform  distribution of parasites by species in the total sample, while all three indices (Shannon, Simpson and BergerParker)  showed  that  the  area  of  Bačka  had  the  highest biodiversity  of  helminth  community.  For  the  species Echinococcus multilocularis and  Alaria  alata  was confirmed a statistically significant difference in the percentage of infected foxes  depending  on  the  area  of  Vojvodina.  According  to  the taxonomic structure of helminth fauna, in foxes was proven the highest percentage of mixed infection (61%), while the number of species per host ranged from 1 to 5. There were registered 12 different combinations of parasite species. In jackals, the mixed infection was also the most common  (59%), while the number of  species  per  host  ranged  from  1  to  4.  There  were  also registered  11  combinations  of  intestinal  parasites.  The coprologic test in this study proved to be low sensitiv and less specific  for  identification  of  parasites  compared  to  direct examination  of  the  intestine  after  autopsy  of  the  animal.  The prevalence of infested foxes (56.3%) and jackals (48%) proven by  the  coprological  method  was  lower  compered  to  the prevalence of patological finding. The statistically significant influence  of  sex  on  the  prevalence  of  infection  was  not registered in neither one of the host species. Of the identified species  of  helminths,  at  foxes  and  jackals,  more  than  half  of them  have  zoonotic  or  potential  zoonotic  significance:  Alaria alata,  Mesocestoides  spp.,  Echinococcus  multilocularis, Toxocara canis  i  Uncinaria stenocephala.  The most important among  them  is  the  tapeworm  E.  multilocularis,  the  cause  of echinococcosis, a disease classified as a zoonosis of priorities in Serbia. In the area of Srem, the occurence of foci of multilocular ecinococcosis  was  proved,  which  is  why  it  is  necessary  to optimize the strategy to control the spread of parasites

Atypical antipsychotic clozapine binds fibrinogen and affects fibrin formation

Clozapine is an atypical antipsychotic used for the treatment of schizophrenia. The prescribed target daily doses may reach 900 mg. Literature studies report a connection between clozapine usage and thrombosis development. Our in vitro study aimed to provide insight into molecular bases of this observation, investigating clozapine binding to fibrinogen, the main plasma protein involved in hemostasis. Fibrinogen/clozapine interaction was confirmed by protein fluorescence quenching, with an affinity constant of 1.7 × 105 M−1. Direct interactions did not affect the structure of fibrinogen, nor fibrinogen melting temperature. Clozapine binding affected fibrin formation by reducing coagulation speed and thickness of fibrin fibers suggesting that in the presence of clozapine, fibrinogen may acquire thrombogenic characteristics. Although no difference in fibrin gel porosity was detected, other factors present in the blood may act synergistically with altered fibrin formation to modify fibrin clot, thus increasing the risk for development of thrombosis in patients on clozapine treatment. ORAC and HORAC assays showed that clozapine reduced free radical-induced oxidation of fibrinogen. All observed effects of clozapine on fibrinogen are dose-dependent, with the effect on fibrin formation being more pronounced.This is the peer-reviewed version of the article: Gligorijević, N.; Vasović, T.; Lević, S. M.; Miljević, Č.; Nedić, O.; Nikolić, M. Atypical Antipsychotic Clozapine Binds Fibrinogen and Affects Fibrin Formation. International Journal of Biological Macromolecules 2020, 154, 142–149. [https://doi.org/10.1016/j.ijbiomac.2020.03.119

FIRST RECORD OF PYRGUS CINARAE (LEPIDOPTERA: HESPERIIDAE) IN SERBIA

Pyrgus cinarae (Rambur, 1839) is currently known from eastern Spain, Albania, Macedonia, Greece, Bulgaria, Ukraine and Russia. At the end of June and beginning of July 2014 it was recorded in Serbia for the first time, expanding its known area of distribution in the Balkan Peninsula northwards. These records point out that southern Serbia may include several additional Mediterranean and Submediterranean species, and call for more systematic study and protection of butterflies in the region. The list of butterfly species recorded in Serbia has increased to a total of 199

B. burgdorferi infekcija u krpeljma uklonjenih sa ljudi i prisustvo antitela protiv borelija kod pacijenata infestiranih krpeljima

The primary objectives of this study were (1) to determine the presence of Borrelia burgdorferi infection in ticks removed from patients for the purpose of singling out sites with increased risk of Lyme borreliosis, and (2) to determine the presence of IgM and/or IgG antibodies against B. burgdorferi sensu lato (s. l.) complex in sera of patients who had ticks removed. From 108 ticks removed from patients, all were examined zoologically and a sub-sample of 91 ticks was tested using PCR analysis to determine the presence of DNA indicating B. burgdorferi infection. To detect anti-Borrelia IgM and/ or IgG antibodies in 61 patients bitten by ticks, we used line recombinant immunoblot test. The most common tick identified was Ixodes ricinus. B. burgdorferi s. l. was present in 37 of 91 tested ticks (40.7%). Seroconversion against B. burgdorferi s. l. antigen was detected in 12 of 61 patients (19.7%). Most of the infected ticks were from the province of Vojvodina (11 municipalities), with the city of Novi Sad proving to be the site with the highest number of infected ticks, 6 in total.Glavni cilj ove studije je bio da se ispita prisustvo B. burgdorferi infekcije u krpeljma uklonjenih sa pacijenata radi utvđivanja lokaliteta sa povećanim rizikom za obolevanje od lajm borelioze, kao i ispitivanje prisustva IgM i/ili IgG antitela usmerenih protiv B. burgdorferi s. l. kompleksa u serumu pacijenata infestiranih krpeljima. Sa pacijenata je prikupljeno i zoološki ispitano 108 krpelja. PCR analiza krpelja korišc'ena je za utvrđivanje prisustva B. burgdorferi s. l. kompleks kod 91 krpelja od ukupnih 108. Za otkrivanje IgM i/ili IgG antitela protiv B. burgdorferi s. l. korišćeni su linijski rekombinantni imunoblot testovi. Najčešc'i identifikovani krpelj je Ixodes ricinus. B. burgdorferi s. l. je bila prisutna u 37 od 91 testiranog krpelja (40,7%). Prisustvo IgM/IgG antitela protiv B. burgdorferi s. l. otkriveno je kod 12 od 61 pacijenta (19,7%). Najviše inficiarnih krpelja je sa teritorije AP Vojvodine (11 opština), gde je urbani deo Novog Sada lokalitet sa najvećim brojem inficiranih krpelja - 6

Atypical antipsychotic clozapine binds fibrinogen and affects fibrin formation

Clozapine is an atypical antipsychotic used for the treatment of schizophrenia. Prescribed daily doses of clozapine may reach over 900 mg/day. Some studies reported a connection between clozapine usage and the development of thrombosis. Our in vitro study aimed to provide insight into molecular bases of this observation, investigating clozapine binding to isolated fibrinogen, the main protein involved in hemostasis. Fibrinogen/clozapine interaction was confirmed by protein fluorescence quenching, with affinity constant calculated to be 1.7 9 105 M1 and the number of binding sites more than one. Direct interactions do not affect the structure of fibrinogen, as determined by UV-VIS spectrometry and Fourier-transform infrared spectroscopy, nor fibrinogen melting temperature, examined by fluorescence spectroscopy. However, clozapine binding affected fibrin formation, by reducing coagulation speed and thickness of fibrin fibers. This behavior suggests that in the presence of clozapine, fibrinogen may acquire thrombogenic characteristics. Although no difference in fibrin gel porosity was detected, other factors present in the blood may act synergistically with altered fibrin formation to modify fibrin clot, thus increasing the risk for development of thrombosis in individuals on clozapine treatment. By ORAC and HORAC antioxidant assays, we found that clozapine efficiently protects fibrinogen from free-radicals oxidation. Since the effect of clozapine on fibrin formation is dose-dependent, it seems that the dosage of the medication could be the main factor that determines if clozapine will have a more positive or negative effect on fibrinogen and coagulation process in vivo

Supplementary data for the article: Šunderić, M.; Vasović, T.; Milčić, M.; Miljević, Č.; Nedić, O.; Nikolić, M. R.; Gligorijević, N. Antipsychotic Clozapine Binding to Alpha-2-Macroglobulin Protects Interacting Partners against Oxidation and Preserves the Anti-Proteinase Activity of the Protein. International Journal of Biological Macromolecules 2021, 183, 502–512. https://doi.org/10.1016/j.ijbiomac.2021.04.155.

Supplementary material for: [https://doi.org/10.1016/j.ijbiomac.2021.04.155]Related to published version: [https://cherry.chem.bg.ac.rs/handle/123456789/4538

Uranium contents from bomb craters

Depleted uranium (DU) is a by-product of uranium enrichment process and its use is very dangerous and harmful. We measured uranium concentration in the surface soil (0-5 cm depth) from bomb craters caused by cruise missiles in which DU might have been used as counterweights during NATO attack. Total uranium concentration and isotopic ratio were determined using y-spectrometry and inductively coupled plasma method. Obtained results were in the range 1.7 - 22 mg/kg dry soil that was comparable to the uranium values found in off-side locations of Serbian soils

Human Hemoglobin and Antipsychotics Clozapine, Ziprasidone and Sertindole: Friends or Foes?

Packed with hemoglobin, an essential protein for oxygen transport, human erythrocytes are a suitable model system for testing the pleiotropic effects of lipophilic drugs. Our study investigated the interaction between antipsychotic drugs clozapine, ziprasidone, sertindole, and human hemoglobin under simulated physiological conditions. Analysis of protein fluorescence quenching at different temperatures and data obtained from the van’t Hoff diagram and molecular docking indicate that the interactions are static and that the tetrameric human hemoglobin has one binding site for all drugs in the central cavity near αβ interfaces and is dominantly mediated through hydrophobic forces. The association constants were lower-moderate strength (~10^4 M^-1), the highest observed for clozapine (2.2 x 10^4 M^-1 at 25 °C). The clozapine binding showed “friendly” effects: increased α-helical content, a higher melting point, and protein protection from free radical-mediated oxidation. On the other hand, bound ziprasidone and sertindole had a slightly pro-oxidative effect, increasing ferrihemoglobin content, a possible “foe”. Since the interaction of proteins with drugs plays a vital role in their pharmacokinetic and pharmacodynamic properties, the physiological significance of the obtained findings is briefly discussed