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Between the years 2000 and 2004, 93,107 sera from 1,997 pig herds in 11 regions of Croatia were tested for the presence of antibodies against brucellosis. Positive results were observed in 67 herds from seven regions (mean individual prevalence: approximately 1%; herd prevalence: 3.4%). The herds from all but two of the infected farms were reared outdoors and thus almost certainly came into contact with wildlife. From 2003 to 2004, 424 sera, which were randomly collected from hunted wild boar (Sus scrofa), were also tested and shown to have a mean seroprevalence of 27.6%. Brucella was isolated from 88 out of 151 serologically positive pigs (58.3%) and 7 of the 93 (7.5%) wild boar which were randomly submitted for bacteriological study. All but three isolates were Brucella suis biovar 2; the others being biovar 3. These results suggest that brucellosis is enzootic in Croatian populations of wild boar. These populations represent a potential disease reservoir for free-range pig farms, as they do in other countries of Central and Western Europe. This is the first report of B. suis biovar 3 in swine and wild boar in Europe, which is an issue of serious concern for public health.

During the 10-year survey (1996–2005), a total of 868 blood samples from different game species in Croatia were analyzed for the presence of leptospiral antibodies. The specific antibodies (AB) were detected in 242 samples (27.88%). According to the species in red deer (Cervus elaphus), the antibodies against six different leptospiral serovars were found in 43 of 226 analyzed sera (19.02%). The most frequent antigen serovars in the deer population were Pomona and Ballum (with the same frequency of 23.6%), whereas the highest titer was recorded for serovar Sejroe (1:800). In the analyzed roe deer (Capreolus capreolus) serum samples, a low level of leptospiral antibodies (6.07%) was determined, with just two AB for antigen serovars—Australis and Sejroe. In wild boar (Sus scrofa), leptospiral antibodies were detected in 151 of 431 samples analyzed (35.03%), with AB for nine antigen serovars. The serovars most frequently found were Australis (48.70%) and Pomona (22.70%), and these serovars also recorded the highest titer (1:3,200). Among brown bear (Ursus arctos) samples, leptospiral antibodies were detected in 25.00% of the samples, with four AB for antigen serovars, of which the most frequent was Icterohaemorrhagiae (>40%). This serovar had the highest recorded titer (1:400). From 112 analyzed red fox (Vulpes vulpes) samples, leptospiral antibodies were found in 35 samples (31.25%). The determined antibodies were specific for four antigen serovars, of which the most frequent (46.2%) and with highest titer (1:1600) was serovar Australis. No antibodies (28/0) were recorded in mouflon (Ovis musimon). The most important game species from an epizootiological point of view in the studied area were certainly wild boar and red foxes. With strong serological reactions, these two species could be emphasized as important hosts for Leptospira interrogans sv. Australis in Croatia, but for their declaration as ‘maintaining hosts,’ isolation of sv. Australis is needed. According to aerial distribution, the highest number of positive samples from different game species was recorded in the central and eastern parts of Croatia, known as the ‘historical natural foci’ of leptospirosis—the regions of Posavina, Podravina, Slavonija, and Baranja. In contrast, the areas of Kordun and Gorski Kotar are declared as leptospira low-risk regions for the game species studied.

During the years 2001 and 2002 on seven localities in Croatia a survey on the prevalence of brucellosis in wild boar was carried out. The survey included 271 (52, 7%) female and 243 (47, 3%) male animals between 7 months and 4 years of age and weighing from 14 to 135 kg. On that occasion 514 blood samples of wild boar were serologically analysed. For serological analysis indirect enzyme immunoassay (iELISA), Rose Bengal test (RBT), complement fixation test (CFT) and slow agglutination test (SAT) were used. In all of the wild boar from all of the localities investigated positive reactions to brucellosis were established. Most of the positive reactions were established by iELISA (13, 6%), then by RBT (11, 5%), CFT (10, 5%) and SAT (8, 9%). Tissue samples of 106 animals: testes samples from 67 animals, uterus tissue from 38 animals and 5 fetuses of piglets from 1 mother were analysed bacteriologically. Brucella suis biovar 2 was isolated from 18 (17, 0%) animals that originated from all of the localities investigated. Isolates were identified by PCR using BRU-LOW primers specific for genus Brucella and primers specific for IS711. Based on our results it could be concluded that in Croatia wild boar are natural vector and/or reservoirs of B. suis biovar 2. This permanent risk factor is hazardious for domestic and wild animals in the Republic of Croatia.

In total the samples of blood and kidneys of 379 small rodents and 154 wild swine were analysed. The antibodies to different serovars of leptospirosis were determined in 12.7% of small rodents, most often in the species Mus musculus (34.4%), A. agrestis (14.8%), A. flavicolis (10.8%), C. glareolus (9.4%) and A. sylvaticus (6, 5%). Most frequent were the findings of antibodies to sv. pomona (27.1%), sv. sejroe (20.8%) and sv. australis (14.6%)and the antibodies to sv. hardjo, sv. saxkoebing, sv. tarassovi, sv. grippotyphosa, sv. bataviae and sv. icter-haemorrhagiae were also established. Seventeen (4.5%) isolates were identified, belonging to the serogroups sejroe (10 isolates), pomona (4 isolates), and australis (1 isolate) and one isolate was not identified. In wild swine positive reactions were established in 26% of blood sera analysed. Most frequently the antibodies to sv. pomona (47.5%), sv. australis (40%), sv. grippotyphosa (10%) and sv. icterohaemorrhagiae (2.5%) were established. Thirteen (8.4%) isolates belonging to the serogroups pomona (10 isolates), australis (2 isolates) and icterohaemorrhagiae (1 isolate) were identified.

Eastern Croatia (Kopački rit) is one of the most famous habitat of red deer (Cervus elaphus). Knowledge of normal electrolyte and mineral concentrations, together with other hematological and biochemical values, are essential for appropriate diagnosis of diseases and health control. The aim of this study was to determine mineral and electrolyte concentrations in young red deer population and compare them with adults. Serum total calcium (tCa), inorganic phosphorus (iP), total magnesium (tMg), total protein and albumin were determined by spectrophotometric analysis; and ionized calcium(Ca2+), magnesium (Mg2+), sodium (Na+), potassium (K+) and chloride (Cl-) concentrations were determined by ion-selective electrodes (ISE). Young animals showed significantly higher values of K+, tCa, Ca2+, Mg2+, iP and albumin, while Na+, Cland protein concentrations were lower. There was no significant difference in the tMg concentration and Ca2+/Mg2+ ratio in relation to age. In wild animals, these laboratory analyses are still in their very beginning due to difficulties in collecting data and lack of equipment in veterinary practice. Electrolyte and mineral values determined in this study can be considered reference data for health control and disease diagnosis, Istočni dio Hrvatske (osobito područje Parka prirode Kopački rit) značajno je stanište jelena običnog (Cervus elaphus). Utvrđivanje referentnih vrijednosti za laboratorijske pokazatelje kod brojnih vrsta divljih životinja, pa tako i ove, nije još u potpunosti provedeno. Razlog tome su poteškoće u prikupljanju uzoraka i relativno pomanjkanje opreme u područnim laboratorijima. Utvrđivanje referentnih raspona za serumske odnosno plazmatske koncentracije minerala i elektrolita, uz ostale hematološke i biokemijske pokazatelje predstavlja temelj za objektivno procjenjivanje zdravstvenog stanja. Pri tome je posebno važno precizno utvrditi referentne vrijednosti za svaku pojedinu dobnu kategoriju, kako zbog nepoznavanja fizioloških razlika ne bi došlo do pogrešnog tumačenja laboratorijskih nalaza. Cilj ovog istraživanja bio je odrediti i usporediti serumsku odnosno plazmatsku koncentraciju minerala i elektrolita kod mladih i odraslih jedinki u običnog jelena te utvrditi postoje li značajne razlike ili se mogu koristiti iste referentne vrijednosti za ove pokazatelje u obje dobne skupine. Serumska razina ukupnog kalcija (tCa), anorganskog fosfora (iP), ukupnog magnezija (tMg) te albumina i ukupnih proteina određena je spektrofotometrijski, a plazmatska koncentracija ioniziranog kalcija (Ca2+), magnezija (Mg2+), natrija (Na+), kalija (K+) i klorida (Cl-)određena ion-selektivnim elektrodama (ISE). U mladih jedinki nađene su signifikantno više vrijednosti K+, tCa, Ca2+, Mg2+, iP i albumina, dok su vrijednosti Na+, Cl- i proteina bile niže u odnosu na odrasle jelene. Za tMg i omjer Ca2+/Mg2+ nije ustanovljena značajna razlika kod različitih dobnih kategorija. Dobiveni rezultati mogu se smatrati referentnim za koncentraciju istraživanih minerala i elektrolita u jelena običnog (Cervus elaphus) te biti oslonac u procjeni i praćenju zdravstvenog stanja ove divljači. Pri tome treba istaknuti da je za neke pokazatelje utvrđena fiziološka razlika kod različitih dobnih kategorija pa je za pravilno tumačenje nalaza od najvećeg značenja rabiti referentne vrijednosti dobivene od primjerene dobne skupine.

Eastern Croatia (Kopacki rit) is one of the most famous habitat of red deer (Cervus elaphus). Knowledge of normal electrolyte and mineral concentrations, together with other hematological and biochemical values, are essential for appropriate diagnosis of diseases and health control. The aim of this study was to determine mineral and electrolyte concentrations in young red deer population and compare them with adults. Serum total calcium (tCa), inorganic phosphorus (iP), total magnesium (tMg), total protein and albumin were determined by spectrophotometric analysis ; and ionized calcium (Ca2+), magnesium (Mg2+), sodium (Na+), potassium (K+) and chloride (Cl-) concentrations were determined by ion-selective electrodes (ISE). Young animals showed significantly higher values of K+, tCa, Ca2+, Mg2+, iP and albumin, while Na+, Cl- and protein concentrations were lower. There was no significant difference in the tMg concentration and Ca2+/Mg2+ ratio in relation to age. In wild animals, these laboratory analyses are still in their very beginning due to difficulties in collecting data and lack of equipment in veterinary practice. Electrolyte and mineral values determined in this study can be considered reference data for health control and disease diagnosis.

During the research period (2001-2005), the lymph nodes of 123 wild boars from seven locations in the Republic of Croatia were tested for the presence of mycobacteria. Mycobacteria were isolated from the lymph nodes of 15 (12.2%) wild boars from fifi ve locations. Specific hybridization showed that 11 (8.9%) isolates belonged to M. avium, 2 (1.6%) isolates were typed as M. fortuitum, and 2 (1.6%) isolates belonged to M. gordonae. All 11 isolates belonging to M. avium sp. were tested by means of the specififi c primers P1 FR300 and P2 FR300. Product size in all isolates was 300 bp, which confirms that M. avium subsp. hominissuis was present in all of them. As in swine from intensive farming, M. avium subsp. hominissuis is a dominant mycrobacteria type in wild boar in Croatia. The results lead to the conclusion that the most important source for wild boar is an environment suitable for the development and survival of M. avium subsp. hominissuis, as confirmed in earlier research studies by other authors., Tijekom istraživanog razdoblja (2001. do 2005.) radi pretrage na prisutnost mikobakterija obrađeni su limfni čvorovi 123 divlje svinje iz sedam lokaliteta u Republici Hrvatskoj. Mikobakterije su izdvojene iz limfnih čvorova 15 (12,2%) divljih svinja iz pet lokaliteta. Specifičnom hibridizacijom dokazano je da 11 (8,9%) izolata pripada vrsti M. avium sp., 2 (1,6%) izolata tipizirana su kao M. fortuitum, a 2 (1,6%) izolata pripadali su vrsti M. gordonae. Svih 11 izolata koji su pripadali M. avium sp. testirani su pomoću specifičnih početnica P1 FR300 i P2 FR300. U svih izolata veličina proizvoda iznosila je 300 parova baza što potvrđuje da se u svih radi o vrsti M. avium subsp. hominissuis. Kao i u svinja iz intenzivnog uzgoja, M. avium subsp. hominissuis predstavlja dominantnu vrstu mikobakterija u divljih svinja u Hrvatskoj. Na temelju dobivenih rezultata može se zaključiti da je po svemu sudeći najvažniji izvor za divlje svinje okoliš koji pogoduje razvoju i opstanku M. Avium subsp. hominissuis, što potvrđuju i ranija istraživanja drugih autora.

The objective of this study was to investigate the haematological and biochemical values of clinical significance for red deer (Cervus elaphus) and provide data for farmed fawns. Blood samples were collected regularly from 34 fawns and compared with 11 adults. The mean blood haemoglobin (Hb), total erythrocyte count (RBC), packed cell volume (PCV), total leukocyte count (WBC) and percent of segmented neutrophils were signifi cantly lower in fawns. Red distribution width (RDW), total platelet count (PLT), plateletcryt (PCT) and percent of eosinophils and lymphocytes were signifi cantly higher in fawns. A vast majority of biochemical parameters were signifi cantly lower in fawns: blood urea nitrogen (BUN), creatinine (CRE), total protein (tPROT), total bilirubine (tBIL) and glucose (GLU) concentration, as a aspartate-aminotransferase (AST) and creatin phosphokinase (CPK) activity, while albumine concentration (ALB), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT) activity showed a signifi cant increase. These results demonstrate the need to use specifi c reference intervals for deer of different ages. The values reported here can be used as a starting point for reference range establishment for clinically healthy young farmed red deer in Croatia.

Monensin is a monovalent Na+-selective carboxylic polyether ionophore antibiotic, a metabolic product of Streptomyces cinnamonensis. Monensin is effective as growth-promoting agent and is used in beef cattle rations. Toxicity results from exposure to undiluted premixes or from mixing errors. Interference with membrane cation transport allows ionophores to form complexes that traverse cell membranes and cause cell death by perturbing the intracellular ionic homeostasis and destabilizing membranes. Monensin toxicity is particularly evident in cardiac and skeletal muscle cells. Lesions are associated with muscle degeneration, acute cardiac failure or chronic cardiovascular insufficiency. No antidote or specific treatment for ionophore toxicoses exist. Acute onset of illness was reported in a beef cattle herd of 70 animals, lethally affecting 18 bullocks, in which monensin had been incorporated in rations considerably exceeding levels prescribed. Deaths occurred 24 h after feeding and were preceded by scarce premonitory signs or more typically, affected animals developed anorexia, diarrhea, cardiac failure with tachycardia, dyspnea, muscular weakness with restricted limb movement and severe muscular pain due to rhabdomyolysis, recumbency, and most strikingly myoglobinuria leading to death after pulmonary congestion and rhabdomyolysis with renal failure. The pathological findings confirmed the clinical and laboratory myoglobinuria showing a mosaic pattern of striated muscles, acute myodegeneration, necrosis, and intrassarcoplasmic lipid deposits in skeletal muscle. Renal histopathology showed extensive deposition of casts of myoglobin. Marked increase was evident in multiple serum enzymes, creatine kinase reaching 377, 640 U/L (normal < 94 U/L). Infectious diseases and poisoning with cardiotoxins such as selenium were excluded. Although the LD50 of monensin for cattle is not firmly established, and deaths occurred at a non-established and different consumption rate, the history of exposure, development of characteristic clinical changes and postmortem findings were consistent with those expected for monensin. To our knowledge, this is the first report of monensin poisoningin Croatia.

Serologic evidence of exposure to porcine parvovirus (PPV) in the wild boar (Sus scrofa) in Croatia was investigated. Serum samples from 219 wild boars captured during 2003 from 12 different locations in the Republic of Croatia were tested by using a commercial enzyme-linked immunoassay (ELISA) and a hemagglutination inhibition (HI) test. Antibodies to PPV were detected in 91 (41.6%) of tested samples and positive results were detected in wild boar from all sample locations. Adults had a significantly higher prevalence (70%) than juveniles (31%; P

Iznesenim radom po prvi je put obavljeno sveobuhvatno istra`ivanje zdravstvenog i genetičkog statusa divljih svinja koje je obuhvatilo područje 7 županija u Republici Hrvatskoj. Sveukupno je pretražen materijal od 745 životinja, od čega je najveći broj svinja odstrijeljenih u lovu 495 komada, zatim uzoraka krvi prikupljenih od živih životinja 264 komada te lešina svinja nađenih u lovištu 31 komad. Odstrijeljene svinje su razvrstane po dobi, spolu i tjelesnoj masi, a lešine su obra|ene holoptićkom i metoptićkom patoanatomskom pretragom. Materijal prikupljen tijekom postmortalne pretrage, ali i krvni serum dobiven intravitalno, podvrgnuti su laboratorijskim pretragama. Od ukupnog broja pregledanih ženskih životinja 72,8% bilo je gravidno. Pozitivni nalaz na svinjsku kugu pretragom slezena dalo je 4,8% pretraženih životinja, dok je serološkom pretragom na protutijela pozitivno reagiralo 39,3% seruma. Pretragama seruma na parvovirusnu infekciju svinja, PRRS, brucelozu i leptospirozu pozitivan titar dalo je 38%, 10,8%, 14% odnosno 30,6% pretraženih seruma. Od pretraženih limfnih čvorova na mikroorganizme iz skupine mikobakterija, 10,6% kulturelno je dalo pozitivan nalaz. Parazitološkim pretragama ukupno je determinirano 12 vrsta iz koljena nematoda, 1 iz akantocefala, 3 iz razreda trematoda, 2 iz razreda cestoda, 3 vrste iz skupine protozoa, 1 iz skupine uši, 2 iz skupine krpelja i jedna vrsta iz skupine šugaraca. Procjenom fenotipa svih pretraženih životinja, odstrijeljenih, uginulih i onih ulovljenih u lovke, utvr|ena je znatna promjena fenotipskog izra`aja i to u smislu pojave nepoželjnih obilježja domaće svinje. Svi dobiveni rezultati raspravljeni su te su predložene neke nove smjernice., The research in the presented work was conducted in the hunting ground areas involving 7 counties of the Republic of Croatia. In total, the number of 745 animals was investigated, the majority being represented by material of shot animals during the hunt – 495, followed by samples collected from live animals – 264, and cadavers found in hunting grounds – 31. Animals harvested during the hunt were classified according to age and sex, and their body weight was measured. All the carcasses were studied by pathoanatomical means. The material collected during the postmortem examination as well as the sera obtained intra-vitam were submitted to consecutive laboratory tests. Of the entire number of examined female animals 72,8% were found to be pregnant. A positive result for hog cholera was presented in 4,8% of examined spleen specimens while on serological examination 39,3% were found to have titers of antibodies. In serum reaction for morbus Aujeszky 30,4% was presented with positive titer while for parvovirus infection, PRRS, brucellosis and leptospirosis positive titer was found in 38%, 10,8%, 14%, and 30,6% of examined sera, respectively. On lymph node cultural examination for mycobacterium microorganisms 10,6% was presented as positive. By parasitological means a total of 12 species of phylum nematode, 1 species of phylum acanthocephala, 3 from the class of trematode, 2 from the class of cestode, 3 species from the group of protozoa, 1 species of lice, 2 species of ticks and one species of mites were determined. All of the obtained results are discussed and the need of further investigation is proposed.