Relja Beck

According to the Croatian Livestock Centre’ s data (2006) over 100.000 sheep are bred on the islands for the purpose of producing meat (lambs) and milk (cheese). The majority are raised on the islands of Krk, Cres, Rab, Pag and Brac. Autochthonous (island) breeds predominate on these islands while Istrian and crossbreed sheep are represented in smaller numbers. The sheep are kept extensively and feed on the numerous types of Mediterranean plants there. During the winter period, they are fed on hay, concentrats and pelleted alfalfa. The aim of our research was to establish which diseases affect the health of sheep on Croatian islands. The study was carried out on 40 flocks on the islands of Cres, Krk, Rab, Pag and Brac from 2004 to 2007. The following diagnostic methods were used: clinical examination, necropsy, serological blood tests and coprology tests. It was established that they most frequently suffered from parasitical, infectious and digestive system diseases. The following internal parasites were found: gastrointestinal nematodes, Eimeria sp., Protostongylus sp., Dicrocoelium dendriticum, Fasciola hepatica, Paramphistomum sp., Nematodirus sp., Moniezia sp, Haemmonchus contortus. and Trichuris sp.. As far as external parasites are concerned, the presence of Rhipicephalus and Ixodes ticks were established. Invasion of Melophagus ovinus was identified on the island of Rab. The parasites most frequently affect the health of lambs and young sheep. The presence of abscesses caused by puncture wounds was also established in the sheep. Clinical mastitis was diagnosed on all islands indicating the bacteria Staphylococcus aureus as the major causative agent. The following infectious diseases were found: enterotoxemia, actinobacillosis, contagious ecthyma, footrot and the Brucella ovis infection. As far as diseases of the digestive system are concerned, bloat was found, as well as various forms of indigestion, which are a consequence of overfeeding on green grass, figs or acorns. Diarrhoea widely occurs in sheep since they lick sea salt the so-called posolica drifted by a north-eastern wind to the areas where sheep are bred. Most diseases identified in the study can be prevented and controlled by implementing elaborate safety precautions. When choosing precautions, both breeders and veterinarians have to take account of the size of a flock, objective and stage of production, breeding area, epizootiological situation, diseases identified in the flock so far and season.

Background Canine vector-borne diseases (CVBDs) are caused by a wide range of pathogens transmitted by arthropods. They have been an issue of growing importance in recent years; however, there is limited information about the vector-borne pathogens circulating in Portugal. The aim of the present study was to detect canine vector-borne bacteria and protozoa of veterinary and zoonotic importance using molecular methods. Methods One hundred and forty-two dogs from Lisbon, southern Portugal, were tested: 48 dogs from a veterinary hospital clinically suspected of vector-borne diseases and 94 apparently healthy dogs from shelters. Anaplasma spp./Ehrlichia spp., Babesia/Theileria spp., Hepatozoon spp., and Mycoplasma spp. infections were detected by PCR from blood samples and examined under light microscopy. Other information including clinical status and diagnostic test results were collected for each animal. Results Infections were detected by PCR in 48 (33.80%) dogs. Single infections w...

Background The apicomplexan haemoparasite Theileria equi, a causative agent of equine piroplasmosis, is an established pathogen of significant welfare and economic concern within the Croatian equine population. A previous large surveillance study of T. equi has identified two distinct parasite populations, one in the north and one in the south, geographically separated by the Dinaric Alps, which traverse the country. This study aimed to further investigate the genetic diversity within these two populations, focussing on allelic variability of the equi merozoite antigen gene, ema-1. Methods Following nested PCR of DNA isolates, the generated ema-1 amplicons were subsequently sequenced and compared by phylogenetic analysis to available sequences representing previously described ema-1 genotypes (groups A–C). Results Isolates from the southern T. equi population clustered with the existing ema-1 groups A and B. Strikingly, isolates from the northern population clustered into two novel ...

Dr. sc. Iva ŠMIT, dr. med. vet., znanstvena novakinja, dr. sc. Dalibor POTOČNJAK*, dr. med. vet., redoviti profesor (dopisni autor, e-mail: dpotocnjak@vef.hr), dr. sc. Marin TORTI, dr. med. vet., docent, dr. sc. Ivana KIŠ, dr. med. vet., docentica, dr. sc. Nada KUČER, dr. med. vet., izvanredna profesorica, dr. sc. Mirna BRKLJAČIĆ, dr. med. vet., docentica, dr. sc. Martina CRNOGAJ, dr. med. vet., docentica, dr. sc. Jelena GOTIĆ, dr. med. vet., viša asistentica, Veterinarski fakultet Sveučilišta u Zagrebu, Hrvatska; dr. sc. Relja BECK, dr. med. vet., znanstveni savjetnik, Hrvatski veterinarski institut, Zagreb, Hrvatska Uvod

... Authors: Tišljar, Marina; Lukač Novak, Irena; Šimpraga, Borka; Krivec, Gabrijela; Savić, Vladimir; Beck, Relja; Marinculić, Albert; Grabarević, Željko; Beck, Ana; Gudan, Andrea; Artuković, Branka; Krstulović, Fani; Balenović, Mirta. ... Made by: Ivo Batistić and Jadranka Stojanovski. ...

Sarkocistoza je bolest koju uzrokuju protoozoe iz porodice Sarcocystidae. Sporulirane oociste i sporociste u izmetu stalnog nositelja mogu se naci koproloskom pretragom. Sarkociste u muskulaturi preživaca i svinja, ako su velike te zahvacene kalcifikacijom, mogu se naci golim okom, a male se nalaze u obojenim histoloskim rezovima , pri cemu debljina ovojnice pomaže u identifikaciji vrste. Zato sto jaca invazija može uzrokovati razlicita patoloska stanja, iako rijetko, i uginuce životinje, ova parazitarna bolest ima sva svojstva znatnog nedostatka. Ako je životinja kupljena za klanje, a sarkocistoza slucajno dokazana u slabijem stupnju i nije izazvala gotovo nikakve organolepticke promjene u mesu, smatrat cemo da sarkocistoza nije znatan nedostatak zato sto se meso zaklane životinje može koristiti bez ogranicenja. Kako su klinicki znakovi rijetki, a nalaz ove bolesti najcesce slucajan, ova bolest ima sva svojstva skrivenog nedostatka. Pojedini merozoiti u jetri, slezeni i limfnim cvorovima mogu se zapaziti 33 dana nakon inokulacije. Transformirajuci merozoiti i shizonti u miofibrilima nalaze se 45 dana nakon inokulacije i okruženi su dvostrukom membranom. Shizonti prve generacije u endotelnim stanicama nastaju 7 dana nakon inokulacije i zavrsavaju razvoj 15 dana nakon inokulacije, a shizonti druge generacije u endotelu kapilara i malih arterija nastaju 19 dana nakon inokulacije. Shizonti se dijele shizogonijom. Od 24 do 46 dana nakon inokulacije mogu se u perifernom krvotoku naci slobodni merozoiti paralelno sa sazrijevanjem druge generacije shizonta.

Mycobacterial spindle cell 'pseudotumour' has been described only once in cats. This unique proliferation of spindle-shaped histiocytes containing Mycobacterium avium is associated with extensive subcutaneous lesions. We report mycobacterial pseudotumour with invasion of muscular and subcutaneous tissues in a 1-year-old female domestic longhair cat. Lesions involved the facial muscles and nasal cavity, making surgical excision impossible. Necropsy examination revealed additional nodules in the subcutis and muscles of the trunk and submandibular lymph nodes. Genotyping of organisms within these lesions revealed simultaneous infection with Mycobacterium avium subsp. avium and Mycobacterium avium subsp. hominissuis. The microscopical appearance of the granulomas was identical, regardless of the strain of bacterium or anatomical location.

Krpelji su, nakon komaraca, najvažniji vektori bolesti ljudi i životinja u svijetu dok su u Europi najznacajniji prenositelji brojnih uzrocnika bolesti. Detaljna i sustavna istraživanja vrsta krpelja i uzrocnika bolesti koje prenose nisu provedena gotovo 50 godina u Republici Hrvatskoj. U ovom istraživanju analizirana su 192 krpelja prikupljena iz okolisa i sa životinja iz Dalmacije sa ciljem dokazivanja vrste krpelja te zoonotskih uzrocnika koje mogu prenijeti. Prisutnost uzrocnika u krpeljima je istraživana lancanom reakcijom polimerazom i neposrednim sekvenciranjem te su dokazane rikecije Rickettsia masillae i R. aeshlimanii iz skupine pjegavih groznica. Morfoloskom determinacijom 192 krpelja dokazane su: Hylomma marginatum (32), Hyalomma scupense (4), Dermacentor marginatus (4), Ixodes ricinus (5), Rhipicephalus bursa (43), Rhipicephalus sanguineus (56) i Rhipicephalus turanicus (48). Sekvenciranjem 16SrRNA potvrđene su sve vrste osim R. sanguineus, stoga se dovodi pitanje postojanje R. sanguineus u Hrvatskoj, a sami rezultati su u suglasju sa novim istraživanjima koji ovog krpelja svrstavaju u skupinu Rhipicephalus sanguineus sensu lato.

Giardia duodenalis is a widespread parasite of mammalian species, including humans. Due to its invariant morphology, investigation on aspects such as host specificity and transmission patterns requires direct genetic characterization of cysts from host samples. The aims of this study were: 1) to determine the intra-assemblage patterns of mutations ; 2) to define multilocus genotypes (MLGs) for assemblages A-F ; and 3) to compare MLGs of human and animal origin in order to estimate the occurrence of zoonotic transmission. A systematic analysis of G. duodenalis isolates from human (n=76) and animal (n=120) was performed by using PCR amplification and direct sequencing of 4 different genes. We found that, with the exception of the small subunit ribosomal RNA gene, the β -giardin (BG), glutamate dehydrogenase (gdh), and triose phosphate isomerase (tpi) genes display high levels of intra-assemblage variability, and define a large number of subtypes, which can be combined to identify MLGs for each assemblage. However, direct sequencing of PCR products also revealed high levels of sequence heterogeneity (the presence of 2 nucleotides at the same position, or mixed positions), a finding that was recently reported by other researchers. This phenomenon appears to be nonrandom with respect to G. duodenalis assemblages. Indeed, mixed positions were absent in all sequences from assemblage A isolates, and, therefore, MLGs can be defined unambiguously. Interestingly, comparison of assemblage A MLGs of human and animal origin showed little support for zoonotic transmission. In assemblage E, mixed positions were detected in all genes, but their number was small (2/11 in BG, 1/12 in tpi, and 3/9 in gdh). The number of isolates of assemblage F tested was too small to draw firm conclusions, albeit mixed positions were rarely observed. These results suggest that MLGs can be determined in the phylogenetically related assemblages A, E and F. On the contrary, in isolates from assemblage B, C and D, mixed positions were more frequently scored (for example, 10/26 in BG, 12/50 in tpi and 15/36 in gdh, among assemblage B isolates). Importantly, isolates that show mixed positions at one gene also show the same feature at the other genes (with some exceptions, however). This makes quite difficult the exact identification of MLGs in those assemblages, and has particularly important implications for genotyping studies of assemblage B isolates. The mechanisms potentially involved in this phenomenon (i.e., true mixed infections, allelic sequence heterogeneity, retention of ancestral polymorphisms and introgression) will be discussed.

Leptospiroza i babezioza pasa mogu pokrenuti fenomen septickog soka koji najcesce zavrsava uginucem. Bolesti se međusobno tesko razlikuju sa klinickog i sa patoloskog aspekta, stoga su nužne mikroskopske, molekularne i seroloske pretrage da se potvrdi je li febrilnu hemoliticku bolest kompliciranu sa septickim sokom pokrenula infekcija protozoonom B. canis ili nekim od serovara bakterije L. interogans. Cilj ovog rada bio je istražiti patoanatomske i histopatoloske podudarnosti i razlike usporedbom nalaza između pet pasa uginulih od babezioze i pet pasa uginulih od leptospiroze. U patoanatomskom nalazu obje skupine hepatomegalija uz edem pluca najcesce je utvrđeni nalaz. Difuzno nekongestivno povecanje slezene neizostavno je u babeziozi dok je renomegalija izazvana akutnom kortikalnom nekrozom potvrđena u svih leptospiroznih pasa. Anemija i blagi ikterus izraženije su u babezioznoj skupini, dok je ikterus jaceg intenziteta uz izostanak anemije potvrđen u leptospiroznoj skupini. U makroskopskom nalazu babezioznih pasa dominirao je edem uz transudaciju u tjelesne supljine dok je leptospirozna skupina u ovom istraživanju pokazala vecu sklonost poremecajima koagulacije sa popratnim krvarenjima. Mikroskopski se babezioza razlikuje od leptospiroze po nalazu merozoita B. canis u eritrocitima, ukoliko psi nisu lijeceni. Također je u babezioznih pasa dokazana znacajnija intravaskularna monocitno- histiocitna leukocitostaza te naglasenija ekstravaskularna hemoliza. Leptospirozni psi u bubrezima razvijaju karakteristicni limfocitno- histiocitni infiltrat na prijelazu kore u srž. Sve ostale lezije u organima vrlo su slicne i temelje se na promjenama u perifernoj redistribuciji krvi uslijed septicnog soka i posljedicnim razvojem hipoksicne nekroze te pojavi diseminirane intravaskularne koagulopatije i krvarenja.

A total of 294 faecal samples were collected during a 1 year period from animal species housed at the Zagreb Zoo, and tested for the presence of Giardia spp. cysts using an immunofluorescence (IF) assay. Giardia spp. cysts were found in the faeces of Primates (Pan troglodytes, Lemur catta), Carnivora (Acinonyx jubatus, Panthera leo, P. uncia, P. pardus, Chrysocyon brachyurus, Helarctos malayanus, Procyon lotor), Artiodactyla (Bison bonasusr, Pecari tajacu, Syncerus cafer), Hyracoidea (Provacia capensis), Pinnipedia (Halichoerus grypus), Rodentia (Callosciurus prevosti), Marsupialia (Trichosurus vulpecula), and Squamata (Elaphe quatrolienata, Eublepharis macularius). In general, the number of cysts was quite small (with the exception of the raccoon ), therefore the possibility of a passive carriage could not be excluded. DNA was extracted from IF positive samples (n= 26), but successful amplification was obtained only for 20 of them. The Giardia species/assemblages were determined by nested PCR and RFLP and/or -giardin (BG) and triose phosphateβ sequence analysis of fragments of the isomerase (tpi) genes. All samples were typed as Giardia duodenalis. TPI sequencing showed the presence of assemblage B in a squirrel and a rock hyrax (subtype B IV), in a chimpanzee and a bear (a new B subtype), and in the lemur (another B subtype), whereas assemblage A has been found in a pecari and in a raccoon (subtypes closely related to A I). Interestingly, assemblage C was identified in a leopard geko (by sequencing) and in a snow leopard (using PCR-RFLP). Furthermore, PCR-RFLP indicated the presence of assemblage B in faecal samples of a lion and a wolf. BG sequencing showed the presence of assemblage A in a oryx sample and in a second pecari sample (both typed as subtype A2, which is of zoonotic relevance). However, in some cases, the two assays have given contrasting results, and the exact identification of the assemblages was problematic. These samples are currently being analysed using other PCR assays, in an effort to understand the causes of this assemblage “ swapping” . This is the first report on the molecular characterization of Giardia duodenalis from Zoo animals, and has revealed many potential new host species of this widespread parasite.

... Journal. Authors: Beck, Ana; Beck, Relja; Vrkić, Vanja; Ivan, Conrado, Šoštarić, Zuckermann; Marko, Hohšteter; Artuković, Branka; Janicki, Zdravko; Konjević, Dean; Marinculić, Albert;Grabarević, Željko. ... IRB. Made by: Ivo Batistić and Jadranka Stojanovski. Design: Studio8. ...

Sarkocistoze su parazitske bolesti domacih i divljih životinja uzrokovane protozoama iz porodice Sarcocystidae. Nalaz sarkocista u misicju srnece divljaci opisivalo je vise autora. a postotak nalaza ovih cista u misicju srna krece se oko 70%. Iako veliki broj srnece divljaci služi kao posrednik u razvoju ovog parazita, nalaz klinickih znakova pa tako i patohistoloskih promjena u invadiranome misicju je razmjerno rijedak. Između ostaloga u radu je iznesen i prikaz triju slucajeva sarkocistoze u srcanom misicu srnece divljaci obrađene na Veterinarskom fakultetu Sveucilista u Zagrebu. Preventivne mjere u suzbijanju sarkocistoze provode se u konacnih nositelja.

ABSTRACT A reliable epidemiological data regarding the prevalence and species differentiation of coccidian parasites in sheep-farming operations in the North Dalmatia (Croatia) is lacking. The aim of this study was to collected the data and analyze the epidemiological status of ovine coccidiosis in this coastal region of Croatia, and to identify the species involved. Fecal analysis revealed the presence of the coccidian parasites of the genus Eimeria in four sheep flocks farmed in the Zadar County (North Dalmatia - Croatia). Species differentiation identified 9 different Eimeria species. Of the highly pathogenic species, E. crandallis was found in three sheep flocks, and E. ovinoidalis was present in one. Among the other pathogenic coccidial organisms, E. parva and E. faurei were found in two herds, while E. bakunensis was found in one. Non-pathogenic species identified included E. granulosa, E. intricata, E. marsica, and E. pallida. The fecal analysis also revealed a combined presence of five different Eimeria species in two herds, and four different species in other two herds. The epidemiological analysis of 49 lambs indicated that 75.5% of them were shedding coccidian oocysts in feces. Our results show a high prevalence of several different Eimeria spp. in sheep flocks farmed in the North Dalmatia. The presence of several pathogenic species raises a concern for development of clinical coccidiosis in large number of lambs and its potential economic impact for the farmer. These data indicate a need to develop appropriate antiparasitic control programs applicable to the sheep farming practices specific for this Mediterranean region. Key words: coccidia, Eimeria spp., sheep, coastal Croatia

Sažetak: Sarkocistoze su parazitske bolesti domaćih i divljih životinja uzrokovane protozoama iz porodice Sarcocystidae. Nalaz sarkocista u mišićju srneće divljači opisivalo je više autora. a postotak nalaza ovih cista u mišićju srna kreće se oko 70%. Iako veliki ...

During the four year period (2001-2004) a coprological monitoring was applied on free-ranging red deer population at eastern Croatia, with purpose to detect presence of trematodes’ developmental stages in faeces. A total number of 2017 samples were analyzed in laboratory by standard flotation and sedimentation method and at the same time eggs per gram (EPG) determination were done. Eggs of three trematode species were detected, i.e. large American liver fluke (Fascioloides magna), liver fluke (Fasciola hepatica) and ruminal trematode (Paramphistomum cervi), with differences in respect to each hunting ground. Correlation between antitrematod treatment and EPG values for all found trematodes is also given in this paper.

Introduction: Babesiosis, caused by several Babesia species, is one of the commonest infectious diseases of dog populations in Croatia. This intraerythrocytic 2, 4-3 x 4-5 μm teardrop shaped protozoan parasite causing malaria-like disease in dogs is well-known amongst veterinary practitioners for more than 50 years. Prevalence of Babesia canis in continental Croatia is connected with specific tick vector Dermacentor reticulatus distribution. After transmission, piriform merozoites are multiplied within erythrocytes causing febrile haemolytic disease. Uncomplicated babesiosis is characterized with mild to moderate haemolytic crisis dependent on parasitemia levels and erythrocytes destruction degree, although some dogs may be asymptomatic. Symptoms of complicated babesiosis except hypoxic and toxic (haemoglobin, bilirubin, and myoglobin) lesions are related to multiple organs dysfunction syndrome development due to overwhelming systemic inflammatory reaction. Paucity information is available about impact of Babesia rossi on the canine myocardium. Babesia rossi is so far considerated as the most pathogenic subspecies of Babesia known to cause cardiac babesiosis in infected dogs form endemic regions of the South Africa. Myocardial lesion descriptions, both clinical and pathological in Babesia canis infection are entirely lacking. In order to clarify impact of Babesia canis on canine heart, we classified cardiac lesions based on the tissue destruction pattern in 20 dogs. These dogs died or were euthanized due to complicated disease, Babesia canis infection was confirmed by PCR and sequencing. Materials and Methods: Necropsy and histological findings from 7 female and 13 male dogs of different breeds and age, ranging from one month to 12 years were classified based on presence of following macroscopic and microscopic cardiac lesions. Lesions found at necropsy included pericardial effusion, epicardial, endocardial and myocardial haemorrhages and necrosis. Microscopic lesions were identified as presence of endomisial oedema, focal necroses of cardiomyocytes, haemorrhages, sludging of parasitized erythrocytes, extracellular piroplasms presence, intravascular leukocytostasis, micro- thrombi, dystrophic mineralisation and myocarditis. Results: Pericardial effusion were found in 6/20 cases ; 2/6 followed focally extensive infarctions, 4/6 in connection with vascular permeability disturbances connected with epicardial haemorrhages. In 12/20 cases epicardial, endocardial and/or myocardial haemorrhages were found. Histologically, most frequent findings were endomisial oedema (16/20 cases), haemorrhages (13/20 cases) and intravascular leukocytostasis (12/20 cases). Free extracellular piroplasms were present in 7/16 cases with microscopically visible parasite erythrocytes. Sludging of parasitized erythrocytes within myocardial capillaries was found to be possible cause of myocardial haemorrhages in 4/20 dogs ; all of these dogs had pronounced endomisial oedema. Focal necroses were found in 6/20 cases, while extensive neutrophilic and histiocytic myocarditis was found in 4/20 cases as respond to focally extensive myocardial necrosis following thrombosis. Discussion: All dogs in this study found to have myocardial lesions. Most prominent lesions with massive myocardial necrosis are connected with coagulopathy, previously described as a possible complication in Babesia canis infection. All other lesions found are of hypoxia origin, connected with pavementing of parasitised erythrocytes against the endothelium(sludging) or due to plasma dilution and endomisial oedema accumulation as described in cases of infection with Babesia rossi. Our findings suggested that Babesia canis seems to be equally pathogenic as Babesia rossi. Cardiac lesions in clinical cases have to be expected and recognised in cases of complicated babesiosis in Croatian dogs. Babesia canis and Babesia rossi are species specific and infectious only for canines, opposed to Babesia divergnes and Babesia microti confirmed as zoonotic strains of bovine and rodent babesiosis. Human babesiosis is characterised as well as a febrile illness similar to malaria. First human case of babesiosis in the world was diagnosed in Croatia (1957) and since then no cases were recorded. One of possible reason, except low prevalence, is use of automated cell readers in hospitals that cannot detect merozoites in erythrocytes and non- specific clinical signs.

Tick-borne protozoan parasites from the genus Babesia continuously emerge worldwide and are recognized as a substantional public health threat for humans. The main causative agents of human babesiosis in Europe are Babesia divergens, B. divergens-like organism, provisionally designated as EU-1, and recently B. microti. In order to estimate the presence of zoonotic B. microti in Croatia we have analyzed 120 spleen samples from small mouse-like rodents (48 woodmice Apodemus sylvaticus, 37 yellow-necked mice Apodemus flavicolis, 2 striped field mice Apodemus agrarius and 33 bank voles Clethrionomys glareolus) which serve as its reservoir. To determine the role of cervids as potential Babesia reservoirs, spleens of 55 red deer (Cervus elaphus) and a single roe deer (Capreolus capreolus) were also analyzed by PCR and subsequent sequencing of the portion of 18S rDNA. B. microti was detected in 6 yellow-necked mice and 2 bank voles (6.6%). Five of these isolates, detected in both rodent species, had rDNA sequence identical to that of the human Jena/Germany strain (Acc.No. EF413181). The other two isolates were identical to Munich B.microti strain (Acc.No. AB071177) isolated from Mus musculus. In 63.63 % of deer samples only Theileria sp., 99.6% identical to Spanish isolate 3185/02 (DQ866842) was identified. In the single roe deer Babesia was detected that is 100% identical to B. capreoli (AY726010) from roe deer and B. divergens (AY098643) from reindeer (Rangifer tarandus tarandus). The results of this study indicate the presence of zoonotic B. microti organisms in mouse-like rodents Apodemus flavicolis and Clethrionomys glareolus and, at the same time, the absence of zoonotic B. divergens isolates from cervids.

U radu je prikazana patoanatomska i patohistoloska slika visceralne lismanioze u sivog vuka (Canis lupus) uzrokovane protozoom Leischmania infantum. Lesina vuka, mužjaka, cetiri godine starog pronađena je u selu Trolokve u zaleđu Trogira zamrznuta na - 20°C te dostavljena u Zavod za opcu patologiju i patolosku morfologiju Veterinarskog fakulteta u Zagrebu gdje je obavljena razudba. Uzorci kože, misicja i svih parenhimskih organa su fiksirani u 10% neutralnom formalinu i uklopljeni u parafin, a potom su bojeni hemalaun eozinom i Grocott-methamin srebro metodama. Iz obje femoralne vene izvađena je krv, nacinjeni su razmasci i obojeni Diff Quick® ; ; ; ; -om (Shendon) za citolosku pretragu, a sterilno izvađena krv iz srca za seroloske parazitoloske pretrage Dot-ELISA i IFAT. Lijevi preskapularni limfni cvor pohranjen je u sterilni PBS za PCR pretragu. Razudbom je utvrđena kaheksija, generalizirana hipotrihoza s mjestimicnom alopecijom te difuzno, suho, perutanje kože, erozije i ulceracije mekusi i kože u podrucju lijevog kuka, izrazito povecanje svih potkožnih limfnih cvorova i jetre, slabije povecanje slezene, dilatacija srca, jaki edem pluca s konsolidacijom u podrucju desnog apikalnog režnja, nefroza oba bubrega, hipertrofija nadbubrežnih žlijezda i atrofija testisa. Histopatoloskim pregledom limfnih cvorova i slezene utvrđena je atrofija bijele pulpe slezene te smanjenje zone T limfocita u folikulima limfnih cvorova. U svim pregledanim tkivima utvrđena je kronicna mononuklearna upala lokalizirana pretežito oko krvnih žila te u intersticiju organa, a u makrofagima kože, limfnih cvorova, slezene, jetre, nadbubrežnih žljezda i testisa nađeni su amastigotni oblici Leismania sp. U razmascima femoralne krvi također su utvrđeni amastigoti Leischmania sp. promjera 5µ ; ; ; ; m unutar makrofaga i slobodni u krvi. Dot-ELISA dala je pozitivan rezultat u razrjeđenju seruma 1:2560, a IFAT 1:1280. Nested PCR-om utvrđeno je da se radi o Leishmania infantum. U radu se opisuje prvi slucaj lismanioze kao uzroka uginuca u sivog vuka (Canis lupus) u svijetu. Nas nalaz u vuka u potpunosti odgovara slici opisanoj u pasa sto ne iznenađuje s obzirom na filogenetsku srodnost ovih vrsta.. Endemsko podrucje lismanioze i južna stanista vuka u Hrvatskoj se geografski preklapaju pa je cilj ovog rada osim prikaza patoanatomske i patohistoloske slike bolesti skrenuti pozornost na mogucu ulogu vuka u silvaticnom održavanju Leishmania infantum u nasoj zemlji.

The epidemiology of human giardiasis is complex and the relative contribution of antroponotic and zoonotic transmission is still unclear. The occurrence of zoonotic genotypes of Giardia duodenalis in livestock, pets and wildlife is well established, but conclusive evidences of transmission to humans, either directly or indirectly through water or food contamination, are still lacking. Furthermore, molecular investigations have been frequently based on the analysis of a single genetic marker, an approach that may result in an incorrect identification of species/genotypes. In this study, we investigated how reliable is the identification of genotypes as a function of the genetic markers and/or the origin of the parasite (host). The markers chosen were fragments of the genes encoding for the small subunit ribosomal DNA, b-giardin, glutamate dehydrogenase, triose phosphate isomerase and ORF-C4, which were PCR amplified and sequenced. The G. duodenalis isolates were faecal samples from humans, pets, livestock, rodents and wildlife (n=150). A consistent and reliable assignment of genotypes was observed for the human isolates using the 5 markers, and multilocus genotypes were identified for both assemblages A and B. On the other hand, the characterization of animal isolates yielded a different and more complex picture. In particular, the genotype assignment of Giardia isolates from dogs was problematic, as different markers gave different results in about 30% of cases. A similar result was observed for isolates collected from other hosts, albeit at a lower frequency (5-10%). The practical implications of these results, as well as the possible mechanisms that are responsible for, will be discussed.

Giardia duodenalis is a widespread parasite of mammalian species, including humans. Due to its invariant morphology, investigation on aspects such as host specificity and transmission patterns requires a direct genetic characterization of cysts/trophozoites from host samples. A number of molecular assays have been developed to target fragments of the small subunit ribosomal RNA, the  -giardin, the glutamate dehydrogenase, the elongation factor 1-alpha, and the triose phosphate isomerase genes. Recently, molecular epidemiologic approaches have been complicated by the recognition of intra-isolate sequence heterogeneity (i.e., “ mixed templates” ) that affects subtype identification, and of “ assemblage swapping” , that affects assignment of isolates to specific G. duodenalis assemblages. This raises doubts on the way genotyping data have been interpreted so far, and indicates the need to understand the mechanisms that are responsible for these unexpected findings. Here, we critically review those different mechanisms, and discuss some possible experimental strategies that can be used in future studies.

Introduction Dirofilaria repens is a vector-borne filaroid helminth of carnivorous animals, primarily domesticated dogs. Humans are considered to be accidental hosts in which D. repens rarely reach sexual maturity but induce local inflammation, mainly in subcutaneous and ocular tissues. Methods In the current study, we present the detection of multiple adults of D. repens, endosymbiont Wolbachia sp. and microfilariae by molecular analysis in peripheral tissues and bloodstream of a human host. A subsequent meta-analysis of published literature identified 21 cases of human infection with adult D. repens producing microfilariae. Results Within the study population, there were 13 (59.09%) males, eight (36.36%) females and, in one (4.55%) case, sex was not reported. A total of 11 (50.00%) cases had subcutaneous dirofilariasis, six (27.27%) had ocular dirofiliariasis, with single cases (4.55% each) of genital, mammary, lymphatic and a combination of subcutaneous and pulmonary dirofilariasis described. In one (4.55%) case, the primary anatomical site of adult D. repens could not be found. D. repens microfilariae were detected in the local tissue (local microfilariasis) in 11 (50.00%) cases and the peripheral blood (microfilaremia) in 11 (50.50%) cases. Final identification of D. repens microfilariae was based on morphological detection in 14 (63.64%) cases, and molecular detection in eight (36.36%) cases. Conclusion The results of this study suggest that humans may act as a final host for D. repens, however its role as a source of D. repens infection is less clear.

Human alveolar echinococcosis (HAE), caused by the metacestode stage of Echinococcus multilocularis, has emerged in many European countries over the last two decades. Here, we report the first data on the new HAE focus with increasing incidence in central Croatia, describe its clinical presentation and outcomes in diagnosed patients, and provide an update on the prevalence and geographic distribution of Echinococcus multilocuaris in red foxes. After the initial case in 2017 from the eastern state border, from 2019 to 2022, five new autochthonous HAE cases were diagnosed, all concentrated in the Bjelovar-Bilogora County (the county incidence in 2019 and 2021: 0.98/105, in 2022: 2.94/105/year; prevalence for 2019–2022: 4.91/105). The age range among four female and two male patients was 37–67 years. The patients’ liver lesions varied in size from 3.1 to 15.5 cm (classification range: P2N0M0–P4N1M0), and one patient had dissemination to the lungs. While there were no fatalities, postop...

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...

Am 17. Dezember 2004 wurden auf der Insel Rab im Umfeld von 50 Meter 16 tote Gansegeier (Gyps fulvus) gefunden. Ein Geier wurde noch lebend geborgen, aber in moribundem Zustand. In diesem Bericht werden die Resultate pathologischer, bakteriologischer, virologischer, parasitologischer und toxikologischer Untersuchung prasentiert, die fur Zwecke des Feststellung der Todesursache der gefundenen Vogel durchgefuhrt wurden. Auf der Grundlage dieser Untersuchungen und auf der Grundlage klinischer Symptome des moribunden Vogels wurde als Todesursache der Gansegeier akute Vergiftung mit Giften aus der Gruppe von Carbofuran festgestellt. In diesem bericht behandeln wir die moglichen Einflusse von Pestiziden auf die Population gefahrdeter Gansegeier in Kroatien.

Raw milk selling from vending machines appeared in Croatia 4-5 years before. Until then milk was bought in the original packaging with declared chemical composition after pasteurization or sterilization process. For milk from vending machines, there is no obligation to declaring the chemical composition of milk. Therefore, this study attempted to determine chemical quality of raw milk (hereinafter milk) from vending machines. The results indicate a large range of monitored chemical parameters.

Tick-borne protozoan parasites from the genus Babesia continuously emerge worldwide and are recognized as a substantional public health threat for humans. The main causative agents of human babesiosis in Europe are Babesia divergens, B. divergens-like organism, provisionally designated as EU-1, and recently B. microti. In order to estimate the presence of zoonotic B. microti in Croatia we have analyzed 120 spleen samples from small mouse-like rodents (48 woodmice Apodemus sylvaticus, 37 yellow-necked mice Apodemus flavicolis, 2 striped field mice Apodemus agrarius and 33 bank voles Clethrionomys glareolus) which serve as its reservoir. To determine the role of cervids as potential Babesia reservoirs, spleens of 55 red deer (Cervus elaphus) and a single roe deer (Capreolus capreolus) were also analyzed by PCR and subsequent sequencing of the portion of 18S rDNA. B. microti was detected in 6 yellow-necked mice and 2 bank voles (6.6%). Five of these isolates, detected in both rodent sp...

During the four year period (2001-2004) a coprological monitoring was applied on free-ranging red deer population at eastern Croatia, with purpose to detect presence of trematodes’ developmental stages in faeces. A total number of 2017 samples were analyzed in laboratory by standard flotation and sedimentation method and at the same time eggs per gram (EPG) determination were done. Eggs of three trematode species were detected, i.e. large American liver fluke (Fascioloides magna), liver fluke (Fasciola hepatica) and ruminal trematode (Paramphistomum cervi), with differences in respect to each hunting ground. Correlation between antitrematod treatment and EPG values for all found trematodes is also given in this paper.

Introduction: Babesiosis, caused by several Babesia species, is one of the commonest infectious diseases of dog populations in Croatia. This intraerythrocytic 2, 4-3 x 4-5 μm teardrop shaped protozoan parasite causing malaria-like disease in dogs is well-known amongst veterinary practitioners for more than 50 years. Prevalence of Babesia canis in continental Croatia is connected with specific tick vector Dermacentor reticulatus distribution. After transmission, piriform merozoites are multiplied within erythrocytes causing febrile haemolytic disease. Uncomplicated babesiosis is characterized with mild to moderate haemolytic crisis dependent on parasitemia levels and erythrocytes destruction degree, although some dogs may be asymptomatic. Symptoms of complicated babesiosis except hypoxic and toxic (haemoglobin, bilirubin, and myoglobin) lesions are related to multiple organs dysfunction syndrome development due to overwhelming systemic inflammatory reaction. Paucity information is av...