Primarni obostrani difuzni B-velikostanični limfom bubrega s metastazama u središnjem živčanom sustavu u smeđe medvjedice (Ursus arctos) držane u zatočeništvu.

We present the first report of primary bilateral renal diffuse large B-cell lymphoma with central nervous system metastases, in a 7 year-old, captive female brown bear. After 7 months of progressive neurological disease, the animal was euthanized. On necropsy, neoplastic nodules with infiltrative growth in the kidneys were found. Neoplastic proliferation of a similar pattern was found compressing the cerebellum and brain stem, and growing through the nervus maxillaris and nervus ophtalmicus. Histology revealed a highly cellular round cell neoplasm, with an infiltrative diffuse growth pattern through the kidneys and nervous parenchyma. On immunohistochemistry, the neoplastic cells stained for the B-cell marker Pax-5. Our report suggests that lymphoma should be considered as a differential diagnosis in bears with a chronic progressive neurological disease.Opisan je prvi slučaj bilateralnoga primarnog difuznog B-velikostaničnog limfoma bubrega s metastazama u tkivo središnjega živčanog sustava u sedmogodišnje smeđe medvjedice držane u zatočeništvu. Životinja je eutanizirana zbog sedmomjesečne progresivne neurološke bolesti. Obdukcija je pokazala postojanje nodularne infiltrativne neoplazme u oba bubrega. Također je utvrđeno neoplastično nodularno bujanje u području malog mozga i moždanog debla koje je dovelo do značajne kompresivne atrofije navedenih struktura te zahvatilo vidne i maksilarne živce. Histološka pretraga pokazala je da su bubrezi i mozak zahvaćeni difuznim infiltrativnim rastom neoplastičnih pojedinačnih okruglih stanica. Visoka imunohistokemijska pozitivnost neoplastičnih stanica na Pax-5 marker dokazala je da je riječ o B-staničnom limfomu. Ovaj nalaz trebao bi doprinijeti uvrštavanju limfoma na listu diferencijalnih dijagnoza u slučaju pojave kronične progresivne bolesti centralnog živčanog sustava u medvjeda

Procjena izloženosti kadmiju, olovu i živi pri konzumaciji slobodne divljači u Hrvatskoj

Free-living game can be an important source of dietary cadmium and lead; the question is whether exposure to these two elements is such that it might cause adverse health effects in the consumers. The aim of this study was to estimate dietary exposure to cadmium, lead, and mercury from free-living big game (fallow deer, roe deer, red deer, wild boar, and brown bear), and to mercury from small game (pheasant and hare), hunted in Croatia from 1990 to 2012. The exposure assessment was based on available literature data and our own measurements of metal levels in the tissues of the game, by taking into account different consumption frequencies (four times a year, once a month and once a week). Exposure was expressed as percentage of (provisional) tolerable weekly intake [(P)TWI] values set by the European Food Safety Authority (EFSA). Consumption of game meat (0.002-0.5 % PTWI) and liver (0.005-6 % PTWI) assumed for the general population (four times a year) does not pose a health risk to consumers from the general population, nor does monthly (0.02-6 % PTWI) and weekly (0.1-24 % PTWI) consumption of game meat. However, because of the high percentage of free-living game liver and kidney samples exceeding the legislative limits for cadmium (2-99 %) and lead (1-82 %), people should keep the consumption of certain game species’ offal as low as possible. Children and pregnant and lactating women should avoid eating game offal altogether. Free-living game liver could be an important source of cadmium if consumed on a monthly basis (3-74 % TWI), and if consumed weekly (11-297 % TWI), it could even give rise to toxicological concern.Procjena izloženosti kadmiju, olovu i živi pri konzumaciji slobodne divljači u Hrvatskoj Slobodna divljač značajan je izvor kadmija i olova u prehrani ljudi te se nameće pitanje je li izloženost ovim metalima tolika da može štetno utjecati na zdravlje potrošača. Procijenili smo izloženost kadmiju, olovu i živi pri konzumaciji velike slobodne divljači (jelen lopatar, srna, obični jelen, divlja svinja i smeđi medvjed) te živi pri konzumaciji male divljači (fazan, zec) koja je ulovljena u Hrvatskoj između 1990. i 2012. godine. Procjena izloženosti temelji se na dostupnim literaturnim podacima i našim rezultatima mjerenja metala u tkivima divljači te različitoj učestalosti konzumacije (četiri puta godišnje, jedanput mjesečno, jedanput tjedno). Izloženost je prikazana kao postotak vrijednosti (privremeno) prihvatljivog tjednog unosa [(P)TWI] koju je postavila Europska agencija za sigurnost hrane (EFSA). Izloženost toksičnim metalima pri rijetkoj konzumaciji (pretpostavljena za opću populaciju) mesa (0,002-0,5 % PTWI) i jetre divljači (0,005-6 % PTWI) ne predstavlja zdravstveni rizik za potrošače, kao ni redovita (0,02-6 % PTWI) i česta (0,1-24 % PTWI) konzumacija mesa divljači. Preporuka je što više smanjiti konzumaciju iznutrica nekih vrsta divljači zbog visokog postotka uzoraka jetre, a posebno bubrega, koji prelaze zakonom propisane maksimalne razine kadmija (2-99 %) i olova (1-82 %). Djeca, trudnice i dojilje trebale bi izbjegavati konzumaciju iznutrica divljači. Jetra divljači može biti značajan dodatni izvor kadmija ako se redovito konzumira (3- 74 % PTWI), a čestom konzumacijom jetre (11-297 % PTWI) može se povećati rizik od štetnih učinaka na zdravlje

Brown bear feeding habits in a poor mast year where supplemental feeding occurs

The diet of free-ranging bears is an important dimension regarding their ecology, affecting their behavior, population structure, and relation with humans. In Croatia, there has been no recent study on the natural food habits of brown bears (Ursus arctos) or the influence of artificial feeding sites on their diet. During 2017, we collected 53 brown bear stomachs from bears in 2 regions of Croatia— Gorski Kotar and Lika—to assess their diet. Plants—Allium ursinum, the Poaceae family, Cornus mas, berries (i.e., Prunus avium, Rubus plicatus), beechnuts (Fagus spp.)—and various plant parts (i.e., dry leaves, buds, conifer needles, and twigs), as well as mushrooms, made up 80% of the percentage of the volume (%V) of all consumed items. Corn (Zea mays) from the feeding sites made up 37% of the bears’ diet (%V), whereas 20% (%V) was meat and 14% (%V) was pome fruits. Scavenged or preyed animal species, such as wild boar (Sus scrofa), horse (Equus caballus), domestic pig (S. scrofa domesticus), cattle (Bos taurus), roe deer (Capreolus capreolus), and small mammals comprised >66% of %FO (frequency of occurrence), but only 20% of %V. Our results showed that food resources (i.e., livestock such as horses, cattle, and pigs, and corn) found at supplemental feeding sites were more frequently chosen by bears than natural food in 2017, a year characterized by almost no beechnut crop. The results showed that subadult bears obtained most food from feeding sites. These 2 patterns suggested that bears may focus on artificial feeding sites to find food in years when natural food sources are depleted, although this should be tested using diet and food availability data collected from several years.info:eu-repo/semantics/publishedVersio

Želučano-crijevni nametnici i prvi nalaz Giardia spp. u slobodno-živućoj populaciji europskoga smeđeg medvjeda (Ursus arctos) u Hrvatskoj.

Ninety-four European brown bear (Ursus arctos) faecal samples, collected in three counties of Croatia, were examined for the presence of gastrointestinal parasites. Five genera were identified, including the nematodes Baylisascaris transfuga and Syngamus sp., and the protozoan enteropathogens Cryptosporidium sp., Eimeria sp., and Giardia sp. Ancylostomatid eggs were also recovered. Cestodes, trematodes and acanthocephalan eggs were absent from all samples. This is the first parasite survey of brown bears in Croatia in thirty years and the first report of Giardia in this species from the region.Devedeset i četiri uzorka izmeta europskoga smeđeg medvjeda (Ursus arctos), sakupljena u tri županije u Hrvatskoj, pregledana su na prisutnost želučano-crijevnih nametnika. Utvrđeno je ukupno 5 rodova, uključujući nematode Baylisascaris transfuga i Syngamus sp., te enteropatogene protozoe Cryptosporidium sp., Eimeria sp. i Giardia sp. Utvrđena su i jaja ankilostomatida dok jaja cestoda, trematoda i akantocefala nisu pronađena. Ovo je prvo parazitološko istraživanje smeđeg medvjeda u Hrvatskoj u posljednjih 30 godina i prvi nalaz Giardia spp. u ove vrste s ovog područja

Kadmij i olovo u uzorcima vučje jetre: optimizacija metode razgradnje uzoraka uz pomoć mikrovalova

A microwave-assisted digestion method for the determination of cadmium (Cd) and lead (Pb) by inductively coupled plasma mass spectrometry (ICP-MS) was optimised on certified reference material (CRM) (bovine liver, BCR-185R) and wolf liver samples. Different factors influencing digestion efficiency (temperature, time, composition of the digestion mixture, sample mass) were tested. Validation included linearity (up to 200 μg L-1 for Cd and Pb), detection (0.003 μg L-1 for Cd and 0.035 μg L-1 for Pb), and quantification (0.008 μg L-1 for Cd and 0.081 μg L-1 for Pb) limits. Good agreement between measured and certified values was achieved in all conditions, with recoveries ranging from 94 % to 111 % for Cd and from 95 % to 105 % for Pb. The precision of the method, expressed as relative standard deviation, was up to 3 % for Cd and 8 % for Pb. The best digestion parameters (260 °C, 30 min, 1 mL HNO3+4 mL H2O, 0.1 g of CRM) based on accuracy and precision were applied on two wolf liver samples to evaluate the need for the predigestion step (freeze-drying) and appropriate mass of the sample. Freeze-drying improved precision and minimising the tissue mass to 0.1 g reduced the matrix effect. Using these optimised digestion conditions, we determined Cd and Pb in 40 wolf livers collected in Croatia, and their medians (0.055 μg g-1 and 0.107 μg g-1, respectively) were in the range of previously reported data for the grey wolf.Certifi cirani referenti materijal (CRM) govedskih jetara (BCR-185R) i uzorci vučjih jetara koristili su se za optimizaciju metode razgradnje uzoraka prije mjerenja kadmija (Cd) i olova (Pb) masenom spektrometrijom induktivno spregnute plazme (ICP-MS). Ispitivani su različiti uvjeti koji utječu na učinkovitost razgradnje (temperatura, vrijeme, sastav otopine za razgradnju, masa uzorka). Validacijom je obuhvaćeno ispitivanje linearnosti (do 200 μg L-1 Cd/Pb), granice detekcije (0,003 μg L-1 za Cd, 0.035 μg L-1 za Pb) i kvantifikacije (0,008 μg L-1 za Cd, 0,081 μg L-1 za Pb). Postignuto je dobro slaganje izmjerenih i certificiranih vrijednosti u svim ispitivanim uvjetima uz izračunati raspon iskorištenja 94 % do 111 % za Cd i 95 % do 105 % za Pb. Najveća relativna standardna devijacija, kao mjera preciznosti, iznosila je 3 % za Cd i 8 % za Pb. Najbolji uvjeti za razgradnju (260 °C, 30 min, 1 mL HNO3+4 mL H2O, 0,2 g CRM), izabrani prema točnosti i preciznosti, primijenjeni su na uzorke vučjih jetara pri procjeni odgovarajuće mase uzorka i potrebe za uvođenjem dodatnog koraka prije razgradnje (liofilizacije). Liofilizacija poboljšava preciznost pa je preporučujemo pri pripremi uzoraka tkiva. Također, najmanja masa uzorka izabrana je ne samo zbog bolje preciznosti nego i zbog najmanjeg utjecaja matrice te potrebne količine tkiva. Medijani Cd (0,055 μg g-1) i Pb (0,107 μg g-1 mokre mase) dobiveni analizom 40 uzoraka vučjih jetara skupljenih u Hrvatskoj bili su u rasponu vrijednosti objavljenih u literaturi za sivog vuka

Adenokarcinom crijeva kod smeđeg medvjeda (Ursus arctos) - prikaz slučaja

This article presents the first case of intestinal adenocarcinoma in a free living, culled 12 year old, European female brown bear (Ursus arctos), with its characteristic macropathological and histopathological manifestations. Necropsy revealed thoracic gunshot injuries (compassionate shot) and the poor physical condition of the animal, with body fat loss and musculature atrophy. An infiltrative mass protruded the jejunal wall with infiltration of the mesentery, visceral and parietal peritoneum, and distal parts of the colon and rectum. The histopathological examination presented an infiltrative, unencapsulated, moderately cellular neoplastic mass, composed of an epithelial cell population, forming irregular nests, papillary and ribbon like structures and small number of tubules, surrounded by an extensive desmoplastic reaction. Within the tumor a small number of cysts filled with mucin were noted. The epithelial neoplastic cells exhibited mild anisocytosis and anisokaryosis, and a low mitotic index. Histochemical Van Gieson staining showed strong positivity in the desmoplastic proliferation. The epithelial tumor cells were positively imunostained with cytokerain, and stromal cells with vimentin. The histological features presented are characteristic of intestinal adenocarcinomas, of the mixed tubular and mucinous type, with a prominent scirrous reaction.Ovaj članak prikazuje prvi dijagnosticirani slučaj adenokarcinoma crijeva kod 12 godina stare ženke odstrijeljenog slobodnoživućeg smeđeg medvjeda (Ursus arctos), s karakterističnim makroskopskim i histopatološkim svojstvima. Obdukcijom je utrvđena ustrijelna rana (samilosni hitac) na prsnome košu, jaka mršavost s nedostatkom masnoga tkiva i atrofijom mišića. Na jejunumu je ustanovljena infiltrativna masa koja je infiltrirala stijenku crijeva te zahvatila mezenterij, visceralni peritoneum te distalne dijelove rektuma i kolona. Histopatološkom je pretragom utvrđena infiltrativna, neinkapsulirana, umjereno stanična neoplastična masa koja je građena od epitelnih stanica koje formiraju nepravilna gnijezda, papilarne i trakaste strukture te manji broj tubula okruženih jakom dezmoplastičnom reakcijom. Unutar tumora nalazio se manji broj cista ispunjenih sa sluzi. Epitelne neoplastične stanice pokazivale su slabiji stupanj anizocitoze i anizokarioze te nizak mitotski indeks. Histokemijsko Van Giesonovo bojenje pokazalo je jaku pozitivnu reakciju u području dezmoplastične proliferacije. Imunohistokemijska je pretraga pokazala pozitivnu reakciju epitelnih tumorskih stanica na citokeratin, a vezivnotkivnih stanica na vimentin. Utvrđena su histopatološka svojstva karakteristična za adenokarcinom crijeva, mješoviti tubularni i cistični oblik s jakom skiroznom reakcijom

Križanje vukova i pasa u Hrvatskoj

Wolf-dog hybridization is considered as one of the main threats for wolf conservation since the admixture and introgression of domestic genes may disrupt local adaptations and threaten the long term survival of wild wolf populations. We investigated the occurrence of wolf-dog hybridization in Croatia by analyzing a panel of 12 autosomal microsatellite markers using Bayesian admixture tests, and assessed its directionality by the use of maternally and paternally inherited markers in combination with morphometric data and morphological features. A systematic analysis of morphologic features and morphometric data was used to rank the studied individuals into either phenotypic wild-type wolves or suspected hybrids. By combining Bayesian assignment results with phenotypic features, we set three thresholds which differentiated wolves from hybrids with maximized hybrid detection and a minimized chance for false positive hybrid identification. On the basis of phenotype, out of 176 wild canids, 157 (89.2%) were categorized as wolves and 19 (10.8%) as suspected hybrids. On the basis of the Bayesian admixture tests and phenotype together, five (2.8 percent) animals were classified as wolf-dog hybrids, four of them as backcrosses with wolves, and one as a backcross with a dog. Mitochondrial DNA suggested that all hybrids originated from the mating of female wolves and male dogs. Two male hybrids had Y chromosome haplotypes common to both wolves and dogs, while the other two had wolf private Y chromosome haplotypes. One wolf had a dog Y-haplotype, indicating a past introgression of dog genes. All hybrids were found in Dalmatia, where wolves settled recently, and where they live close to humans, with a high rate of human-caused mortality. These conditions are considered as favorable for wolf-dog hybridization. However, we found a low hybridization prevalence in Croatia, which is nonetheless expected to persist as long as the conditions favoring its occurrence are met. The ecological, sociological, conservation and management implications of hybrid occurrence are yet to be determined.Križanje vukova i pasa jedna je od glavnih prijetnji očuvanju vukova jer se miješanjem i introgresijom gena domaćih životinja mogu narušiti lokalne prilagodbe te tako ugroziti dugoročni opstanak populacija vukova u prirodi. Istražili smo pojavu križanja vukova i pasa u Hrvatskoj analizom 12 autosomskih mikrosatelitskih markera Bayesian testom primjese te utvrdili smjer hibridizacije određivanjem markera nasljeđivanih i po majčinskoj i po očinskoj liniji, u kombinaciji s određivanjem morfometrijskih i morfoloških svojstava. Sustavna analiza morfoloških svojstava i morfometrijskih podataka korištena je za rangiranje istraživanih jedinki, bilo kao fenotipske divlje tipove vukova bilo kao sumnjive na križanost. Kombiniranjem rezultata Bayesian statistike s fenotipskim obilježjima postavili smo prag za razlikovanje vukova od križanaca, koji povećava vjerojatnost određivanja križanaca uz smanjivanje vjerojatnosti za pogrešno pozitivno određivanje. Temeljem fenotipa, od 176 divljih kanida 157 (89,2 %) bilo je kategorizirano kao vukovi, dok je 19 (10,8 %) bilo svrstano u sumnjive križance. Temeljem Bayesian testa primjese, pet (2,8 %) životinja bilo je svrstano u križance vuka i psa, četiri od njih određena su kao unatražno križani s vukovima, dok je jedan bio unatražno križan sa psom. Mitohondrijska DNK pokazala je da su svi križanci bili rezultat parenja vučice sa psom. Dva su muška križanca na Y-kromosomu imala haplotipove i od vukova i od pasa, dok su druga dva imala Y-kromosomske haplotipove dosad utvrđene samo kod vukova. Jedan je vuk imao pseći Y-kromosomski haplotip što je uputilo na introgresiju psećih gena. Svi su križanci pronađeni u Dalmaciji, gdje su se vukovi nedavno proširili te gdje žive blizu ljudi, s visokom stopom smrtnosti uzrokovane od ljudi. Te se okolnosti inače smatraju pogodujućima za nastanak križanaca vukova i pasa. Ipak, pronađena učestalost pojave križanaca bila je niska, ali i za očekivati je da će se zadržati sve dok postoje uvjeti koji pogoduju nastanku križanaca. Ekološke, sociološke te značajke hibridizacije za zaštitu i upravljanje populacijom vukova tek treba utvrditi

Diversity of Hepatozoon species in wild mammals and ticks in Europe

Background: Hepatozoon spp. are tick-borne parasites causing subclinical to clinical disease in wild and domestic animals. Aim of this study was to determine Hepatozoon prevalence and species distribution among wild mammals and ticks in Europe. Methods: Samples of wild mammals and ticks, originating from Austria, Bosnia and Herzegovina, Croatia, Belgium and the Netherlands, were tested with PCR to amplify a ~ 670-bp fragment of the small subunit ribosomal RNA gene. Results: Of the 2801 mammal samples that were used for this study, 370 (13.2%) tested positive. Hepatozooncanis was detected in samples of 178 animals (3 Artiodactyla, 173 Carnivora, 1 Eulipotyphia, 1 Lagomorpha), H.martis in 125 (3 Artiodactyla, 122 Carnivora), H.sciuri in 13 (all Rodentia), Hepatozoon sp. in 47 (among which Hepatozoon sp. Vole isolate, all Rodentia) and H.ayorgbor in 4 (all Rodentia). Regarding origin, 2.9% (6/208) tested positive from Austria, 2.8% (1/36) from Bosnia and Herzegovina, 14.6% (173/1186) from Croatia and 13.9% (190/1371) from Belgium/the Netherlands. Of the 754 ticks collected, 0.0% (0/35) Hyalomma sp., 16.0% (4/25) Dermacentor spp., 0.0% (0/23) Haemaphysalis spp., 5.3% (24/50) Ixodes and 1.4% (3/221) Rhipicephalus spp. tested positive for Hepatozoon (4.2%; 32/754), most often H.canis (n = 22). Conclusions: Hepatozooncanis is most present in mammals (especially in Carnivora such as gray wolves and golden jackals) and ticks, followed by H.martis, which was found merely in stone martens and pine martens. None of the rodent-associated Hepatozoon spp. were detected in the ticks, suggesting the possible implication of other arthropod species or non-vectorial routes in the transmission cycle of the hemoprotozoans in rodents. Our findings of H.canis in ticks other than R.sanguineus add to the observation that other ticks are also involved in the life cycle of Hepatozoon. Now that presence of Hepatozoon has been demonstrated in red foxes, gray wolves, mustelids and rodents from the Netherlands and/or Belgium, veterinary clinicians should be aware of the possibility of spill-over to domestic animals, such as dogs. Graphical Abstract: [Figure not available: see fulltext.]

Utvrđivanje raspona dobi i reproduktivne aktivnosti smeđih medvjedica (Ursus arctos) iz zatočeništva pomoću morfohistološke pretrage jajnika - kratko priopćenje

The study aimed to determine reproductive span by investigation of the ovarian structures in young and elderly captive brown bear females (Ursus arctos). The ovaries of two 2-year-old females were obtained by ovariectomy and during the necropsies of 31 and 36 year old individuals. All the obtained ovaries were examined macroscopically and histologically. Histological examination of the ovaries of young animals (2+ years) revealed the presence of primordial, primary, secondary and tertiary follicles within the ovarian cortex. One ovary showed a mature corpus luteum, indicating recent ovulation, what is, to our knowledge, the first histological proof of the earliest age of ovulation recorded for captive brown bears. Ovarian atrophy accompanied by the development of multiple cystic subsurface epithelial structures (SES) in the case of the old bears in this study indicates that the ovaries of brown bears share similar degenerative and proliferative patterns with domestic canids. The oldest female had records of successful births at the ages of 26 and 28 years. Both recorded birth events represent one of the latest confirmed occurrences of ovulation, conception and birth amongst brown bears.Cilj istraživanja bio je pregledom građe jajnika utvrditi raspon dobi i reproduktivne aktivnosti u mladih i starih ženki smeđega medvjeda (Ursus arctos) iz zatočeništva. Jajnici su dobiveni ovariektomijom dviju dvogodišnjih ženki i tijekom razudbe 31-godišnje te 36-godišnje ženke. Svi su jajnici pregledani makroskopski i histološki. Histološkom pretragom u mladih su životinja u kori jajnika otkriveni primordijalni, primarni, sekundarni i tercijarni folikuli. U jednom je jajniku dokazano zrelo žuto tijelo (corpus luteum), pokazatelj nedavne ovulacije što je, prema nama dostupnim podacima, prvi zabilježen histološki dokaz najranije ovulacije u smeđih medvjeda u zatočeništvu. Atrofija jajnika povezana sa starenjem i praćena razvojem multiplih cističnih supseroznih epitelnih struktura (SES) u starih medvjeda u ovom istraživanju upućuje na to da se u smeđih medvjeda pojavljuju slične degenerativne i proliferativne promjene kao i u domaćih kanida. Najstarija ženka imala je zabilježene uspješne porođaje u dobi od 26 i 28 godina. Oba porođaja pripadaju najkasnijim potvrđenim pojavama uspješne ovulacije, začeća i porođaja u smeđih medvjeda