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Krajem svibnja 2015. godine u Laboratorij za parazitologiju, Hrvatskog veterinarskog instituta, Zagreb, Hrvatska dostavljena je krv ovna iz okolice Benkovca sa sumnjom na piroplazmozu. Ovan je uz potpornu terapiju vitaminima AD3E i B-kompleksa liječen imidokarbom (Imizol® 0,5 mL s/c) i oksitetraciklinom (Geomicin retard® 5 mL i/m) te je za pet dana prestao pokazivati kliničke znakove bolesti. U razmazu krvi pronađena su periferno smještena intraeritrocitarna tjelešca, a sekvenciranjem odsječka 16S rRNK i MSP4 gena dokazali smo vrstu Anaplasma ovis, dok DNK drugih uzročnika nije bila prisutna. Ovim istraživanjem smo po prvi puta dokazali vrstu A. ovis u Hrvatskoj, ali i u Jugoistočnoj Europi. Od danas priznatih osam vrsta anaplazmi, anaplazmozu malih preživača uzrokuju tri vrste A. phagocytophilum, A. capra i A. ovis od kojih je A. phagocytophilum dokazana u Hrvatskoj. Prema navodima veterinara Veterinarske ambulante Benkovac opisani klinički znaci bolesti su česti u ovaca i ovnova dopremljenih s drugih područja Hrvatska i nikada nisu uočeni u životinja mlađih od pet mjeseci. Treba naglasiti da osim Imizolom veterinari na području Benkovca oboljele ovce uvijek liječe i tetraciklinima što predstavlja i lijek izbora, a oboljele životinje uspješno izliječe ukoliko lijekove apliciraju unutar pet dana od pojave kliničkih znakova bolesti. Vektor je krpelj Rhipicephalus bursa prisutan diljem Republike Hrvatske ujedno pronađen i na oboljelom ovnu. Stoga treba očekivati da je uzročnik prisutan i u drugim dijelovima Hrvatske., In late May 2015, a sample of ram blood with suspected piroplasmosis from the vicinity of Benkovac was delivered to the Laboratory for Parasitology of the Croatian Veterinary Institute in Zagreb, Croatia. Therapy with Imidocarb (Imizol 0.5 mL s/c) and oxytetracycline (Geomycin retard 5 mL i/m) was started immediately, with supportive therapy with vitamins AD3E and B-complex. The health status improved and clinical signs disappeared after five days, and the ram fully recovered. Peripherally located intraerythrocyte bodies were found in the blood smear, and Anaplasma ovis was confirmed by sequencing the 16S rRNA and MSP4 gene, while the DNA of other pathogens was not present. This research proved the presence of A. ovis in Croatia and in Southeast Europe for the first time. Of the eight species of Anaplasma recognized today, small ruminant anaplasmosis is caused by three species, A. phagocytophilum, A. capra and A. ovis, and to date only A. phagocytophilum has been proven in Croatia. According to veterinarians from the Benkovac Veterinary practice, the described clinical signs of the disease are common in sheep and rams introduced from non-endemic areas and have never been observed in animals younger than five months. It should be emphasized that in addition to Imisol, veterinarians in the Benkovac area always treat sick sheep with tetracyclines, the drug of choice for Anaplasma treatment, and sick animals are successfully cured if treatment is administered within five days of the onset of clinical signs. The vector is the tick Rhipicephalus bursa, which is present throughout the Republic of Croatia and was also found on the infected ram, so it is to be expected that the causative agent is also present elsewhere in Croatia.

Invasive alien crayfish threaten freshwater biodiversity and native crayfish fauna. However, the invasion success of an otherwise successful crayfish invader may be compromised by local environmental conditions or presence of local pathogens which can affect invader’s physiology, immune status, health, and fitness. Here we report the occurence of idiopathic necrotizing hepatopancreatitis and changes in the immune response in the signal caryfish, one of the most successful freshwater invaders, in a recently invaded Korana River in Croatia. We sampled and histologically analysed 73 signal crayfish individuals collected in three consecutive years (2018-2020) and used several standard immune parameters (encapsulation response, hemocyte count, phenoloxidaze activity and total prophenoloxidaze) to: i) compare immune response of the signal crayfish along its invasion range, ii) analyze effects of specific predictors (water temperature, crayfish abundance and body condition) on its immune response changes. Our results show very high prevalence of lesions in hepatopancreas of signal crayfish (>91%), with 14% of individuals displaying heavy and 51% moderate necroses and/or inflammation in the tubular and interstitial structures of the organ. Heavy histopathological changes in hepatopancreas were correlated with lower organosomatic condition. Immune response exhibited significant differences along the invasion range of the signal crayfish and was mostly affected by water temperature and population abundance. Obtained results offer a baseline for elucidating the role of immunocompetence in the signal crayfish invasion in the Korana River, while aetiology of necrotizing hepatopancreatitis needs to be further investigated to determine its potential impact on signal crayfish invasion success.

Enquadramento: O quotidiano familiar de uma criança com doença crónica é modificado decorrente das exigências recorrentes da doença. Os irmãos destas crianças podem receber menos atenção, tanto dos pais, quanto da restante família, que podem interessar-se mais pela criança doente e não da mesma forma pelo irmão considerado saudável. Objetivos: Compilar perceções de irmãos de crianças hospitalizadas por doença crónica. Metodologia: Participaram do estudo irmãos de crianças diagnosticadas com doenças crónicas, com idades entre os 8 e os 16 anos. Utilizaram-se entrevistas abertas, que foram gravadas, transcritas e submetidas à análise. Resultados: Apenas 4 irmãos puderam ser entrevistados. Na análise das entrevistas, emergiram 4 categorias de resultados (Enfrentamento da doença do irmão; Reestruturação familiar; Experiências a partir da hospitalização do irmão; e Experiências de exclusão). Conclusão: Os irmãos das crianças com doenças crónicas estão em situação de vulnerabilidade e exclusão, demonstram dificuldades em lidar com as suas perceções e sentimentos, o que mostra a importância de integrar o irmão no processo de enfermagem. Background: The daily family life of a child with chronic disease changes according to the illness’s demands. The siblings of these children may receive less attention from both parents, as well as from the rest of the family, who may focus more on the sick child and not in a similar way on the healthy sibling. Objectives: To compile perceptions of siblings of children hospitalized due to chronic disease. Methodology: Siblings of children diagnosed with chronic diseases, aged between 8 and 16 years, participated in the study. We used open interviews, recording, transcribing, and submitting them to analysis. Results: We were able to interview only four siblings. In the interviews’ analysis, four categories of results (Coping with the sibling’s disease; Family restructuring, Experiences resulting from the sibling’s hospitalization; and Experiences of exclusion) emerged. Conclusion: The siblings of children with chronic diseases live in a situation of vulnerability and exclusion, demonstrate difficulties in dealing with their perceptions and feelings, which proves it is essential to involve the sibling in the nursing process. Marco contextual: La vida diaria familiar de un niño con enfermedad crónica se modifica de acuerdo con las necesidades recurrentes de la enfermedad. Los hermanos de dichos niños pueden recibir menos atención, tanto por parte de los padres como del resto de la familia, quien puede mostrar más interés por el menor enfermo y no tanto por el hermano considerado sano. Objetivos: Recopilar las percepciones de los hermanos de niños hospitalizados por enfermedad crónica. Metodología: Participaron en el estudio hermanos de niños diagnosticados con una enfermedad crónica, con edades comprendidas entre los 8 y los 16 años. Se utilizaron entrevistas abiertas, que se grabaron, transcribieron y sometieron a análisis. Resultados: Solo se pudo entrevistar a 4 hermanos. En el análisis de las entrevistas, surgieron 4 categorías de resultados (Afrontamiento de la enfermedad del hermano; Restructuración familiar; Experiencias a partir de la hospitalización del hermano; y Experiencias de exclusión). Conclusión: Los hermanos de los niños con enfermedades crónicas están en una situación de vulnerabilidad y exclusión, y demostraron dificultades al lidiar con sus percepciones y sentimientos, lo que muestra la importancia de integrar al hermano en el proceso de enfermería.

The microscopic analysis of cytological samples is a simple, fast, cheap and minimally invasive method for diagnosing pathologic changes in cats, used routinely in many veterinary clinics. This research has examined all changes that were diagnosed in cats from cytological samples delivered to the Department of Veterinary Pathology from 2009 to 2018. The diagnosed changes were classified as one of the following categories: “neoplasia”, “inflammation”, “other pathologic changes” or “nondiagnostic samples”, based on the reported criteria for a cytological examination. For each change, breed, age, gender and anatomic location were analysed by using statistics to determine whether these variables were connected to the diagnosed change. The total number of examined samples, which originated from an approximately equal number of males (49%) and females (51%), was 306. The most frequent change was neoplasia (44%), while inflammation (20%), other pathologic changes (18%) and nondiagnostic samples (18%) were detected in a similar number of the cats. Round cell neoplasia was the most frequent neoplasia (33%), followed by mesenchymal (31%) and epithelial neoplasia (28%). The most common breed was domestic short-haired, but there was no connection between the breed and the category. Pathologic changes were diagnosed in altogether 10 organ systems, of which the skin was the most frequent. The average age of the examined cats was 8.4 years (y), with a variation between categories: the average age of the cats with neoplasia was 9.2 y, with inflammation 8.0 y, with other pathologic changes 8.6 y and with nondiagnostic samples 7.7 y. The most frequent type of inflammation was purulent, followed by pyogranulomatous inflammation. Other types of inflammation were diagnosed in a low number of the cats. Reactive hyperplasia of lymph nodes was the most frequent among other diagnosed pathologic changes. In conclusion, the most frequently diagnosed pathologic changes in the cats were neoplasia, which corresponds to literature data. The most frequent neoplasia was round cell neoplasia, which is the most common neoplasia in cats. The skin was the most frequent tissue sampled, probably due to easy accessibility.

Introduction: Apicomplexan hemogregarines of the genus Hepatozoon, described in more than 300 animal species, are transmitted by various hematophagous arthropods. In the current report we provide the first morphological description, pathological changes and molecular characterization of a Hepatozoon sp. from a stone marten. Materials and Methods: A paralyzed young adult wild female stone marten from Western Croatia was euthanized for determination of cause of illness. During necropsy representative tissue samples were collected for histology and molecular analyses. Abscess content was sampled for bacterial culture and ectoparasites for morphological determination. Results: Necropsy revealed cachexia, head and neck abscesses at the site of bite wounds and severe ectoparasitic infestation. Severe emaciation was caused by massive concurrent bacterial and parasitic infections. Cultivation and histology results confirmed β-hemolytic Streptococcus pyogenes and E. coli within multifocal to coalescing subcutaneous purulent lesions. Although no gross lesions were visible in the heart and cervical muscles, microscopic examination revealed numerous Hepatozoon meronts and released merozoites from ruptured meronts associated with granulomatous inflammation. Macrophages often contained 1.17- 3.12 µm round to oval intracytoplasmatic merozoites. Meronts were sporadically found in the liver, spleen and kidneys without an apparent local inflammation. Sequences of 666 bp fragment of 18S rRNA from the spleen, liver, heart and cervical muscle were identical to Hepatozoon “Badger type” (KU198330). Identical Hepatozoon DNA was confirmed with sequencing in Ixodes ricinus ticks from the stone marten, while fleas from the animal were negative. Conclusions: Infection with Hepatozoon sp. from a stone marten showed similar lesions and organ distribution to those described from different Hepatozoon sp. in other marten species.

In Europe, three types of final hosts of Fascioloides magna are recognized. Among wildlife they include red and fallow deer as definitive hosts, wild boar as dead-end host and roe deer, mouflon and chamois as potential aberrant hosts. The course and outcome of infection are highly influenced by the type of the host. In aberrant hosts the fluke does not reach the sexual maturity and continues to migrate through the liver parenchyma without pseudocyst formation, causing an excessive tissue damage and haemorrhages. Such line of events results in the death of the respective animal. In this study we have analysed 45 roe deer livers originating from the area with established fascioloidosis in red deer. Among positive livers with typical gross lesions already described in aberrant hosts, four of livers had completely developed pseudocysts. Each pseudocyst contained two flukes. We did not find any eggs in the pseudocyst content nor in the accompanied faeces. However, grinding of the flukes revealed the presence of the eggs inside the fluke’s reproductive system. This study shows potential change in the host-parasite relationship, and potential adaptation of the fluke on roe deer and vice versa. The ability to form pseudocyst would reduce mortality in roe deer caused by F. magna infection, but could also modify the role of roe deer in the epidemiology of fascioloidosis.

Ciste koje potječu od folikularnog epitela jesu vrećaste kožne strukture ispunjene keratinom, smještene u potkožju ili u dermisu, s kontaktom s epidermisom i bez kontakta. Te su benigne lezije rijedak nalaz u koži pasa, a obilježava ih spor i progresivan koncentrični rast pri kojemu pritiskom oštećuju i zamjenjuju okolno tkivo kože. Postoji više vrsta cista koje se međusobno razlikuju prema vrsti epitela od kojega je građen njihov zid. Sam tip epitela diktira i sastav keratina u cisti stoga se dijele na: infundibularne, istmusne, matrikalne te hibridne ciste koje su građene od sve tri vrste epitela. Klinička važnost folikularnih cisti temelji se na činjenici da narušavaju ljepotu životinje i uzrokuju zabrinutost u vlasnika, jer ih je bez primjene citološke i/ili histološke dijagnostike nemoguće razlikovati od drugih vrsta benignih i malignih novotvorina podrijetlom od folikularnog epitela kao i od ostalih vrsta tumora kože od kojih boluju psi. Nadalje, što je tijek rasta ciste dulji, količina keratina koja se u njoj nakuplja povećava se te doseže kritičnu točku u kojoj dovodi do kompresivne atrofije zida ciste i njegova stanjivanja. Napeta vrećesta struktura ciste s tankim zidom, koja je k tome i prepunjena keratinom, podložna je sponatanim i traumatskim rupturama pri čemu dolazi do izlaska keratina u okolno tkivo. Keratin u dermisu potiče fulminantnu gnojnu, a kasnije i piogranulomatoznu upalnu reakciju koju vlasnici pacijenata najčešće opisuju kao znatnu oteklinu potkožja koja je nastajala tijekom 2 do 3 dana na mjestu manjega potkožnog nodula. Kirurško uklanjanje ciste dovodi do potpunog izlječenja psa i svrstava folikularne ciste u red prognostički najpoželjnijih dermatoloških dijagnoza. Na primjerima dijagnostičkih algoritama, u tri različita slučaja kožnih cisti folikularnog podrijetla, veterinarima praktičarima bit će objašnjene osobitosti građe zida cisti i prikazan njihov sadržaj te specifičnosti citološkog i histopatoloških nalaza. U kratkom teorijskom pregledu dan je osvrt na podrijetlo i etiologiju nastanka ovih rijetkih lezija kože pasa., Follicular cysts are sac-like structures filled with keratin, located in the subcutis or dermis, with or without contact with the epidermis. These benign lesions are a rare finding and are characterised by slow, progressive and concentric growth, causing compression of the adjacent tissue. Microscopic classification is based on the epithelial lining of the cyst and the type of keratin filling the cyst: infundibular, isthmus, matrical and hybrid follicular cysts may be diagnosed. The clinical significance of follicular cysts is disturbance of the exterior of the animal, raising concern in the owners due to the impossibility of distinguishing these lesions from benign and malignant neoplasms of follicular origin, as well as from other tumour types affecting the skin of dogs without cytological and/or histological diagnostics. Further, with the prolonged growth period of the cyst, the quantity of keratin filling the cyst increases, causing compressive atrophy and thinning of the cyst wall. A tense, sac-like cyst is prone to spontaneous and traumatic rupture, with keratin leaking into adjacent tissue.Free keratin in the dermis causes a strong pyogranulomatous inflammatory reaction often described by pet owners as a fist-sized subcutaneous swelling, developing within 24 to 48 hours at the previous location of a smaller subcutaneous nodule. Surgical removal of the cyst leads to complete cure of the patient, making follicular cysts the most favourable dermatological diagnosis. This paper presents a diagnostic algorithm for follicular cysts based on three different cases of canine follicular cysts, with pictures of gross findings from cut sections of the cyst and cytological and histologic pictures. A brief theoretical overview of the origin and aetiology of these rare lesions in the skin of dogs is also presented.

Fleas and ticks are important vectors of vector-borne pathogens that affect cats and dogs. In the present study ticks and fleas collected from dogs and cats were investigated for presence of vector-borne pathogens and endosymbionts they harbor. The ectoparasites were collected from 14 deceased dogs and 22 deceased cats with gross findings of hemolytic anemia. After the morphological determination of ectoparasites, DNA was extracted for molecular detection of Babesia/Theileria sp., Anaplasma/Ehrlichia sp., Hepatozoon sp., Mycoplasma sp. and Bartonella sp. Ticks Dermacentor reticulatus (4/7, 57%), Ixodes ricinus (4/7, 57%) and Rhipicephalus sanquineus s.l. (1/7, 14%) were collected from 7 out of 14 dogs. Fleas were present in 64% of dogs: Ctenocephalides felis in 67% and C. canis in 33%. In three dogs co-infestation with more than one ectoparasite were detected: (i) C. felis and I. ricinus, (ii) C. canis and D. reticulatus, (iii) D. reticulatus, I. ricinus and R. sanquineus. Babesia canis was detected in four ectoparasites collected from four dogs (4/14, 29%): D. reticulatus (2/4, 50%), I. ricinus (1/4, 25%) and C. canis (1/4, 25%). Rickettsia raoultii was found in a single D. reticulatus (1/14, 7%). Candidatus Midichloria mitochondrii was detected in all collected I. ricinus (4/14, 29%). Two groups of endosymbionts were detected in canine fleas: (i) identical to Wolbachia pipientis, endosymbiont found in C. felis (4/14, 29%) and C. canis (2/14, 14%), (ii) identical to Wolbachia endosymbiont of Xenopsylla sp. found in C. felis (2/14, 14%) and C. canis (1/14, 7%). In two cases co-infections were detected (i) B. canis and W. pipientis in C. canis, (ii) B. canis and Can. Midichloria mitohondrii in I. ricinus. Flea C. felis was present on all examined cats as well as a single I. ricinus tick. Sequences revealed the presence of two endosymbiont groups in C. felis: one identical to W. pipientis (9/22, 41%) and second to W. endosymbiont of Xenopsylla sp. (13/22, 59 %). Anaplasma phagocytophilum was detected in I. ricinus (5%, 1/22), Bartonella clarridgeiae in four (4/6, 67%) and B. hensellae in two fleas (33%). In six cases co-infections were detected (i) W. endosymbiont of Xenopsylla sp. and B. clarridgeiae (4/6, 67%), (ii) W. pipientis with B. hensellae (33%, 2/6).

The lancet liver fluke Dicrocoelium dendriticum occurs in various domestic (sheep, goats, cattle, horses, rabbits, dogs) and wild (deer, wild boar, buffalo, camels) mammals and humans around the globe. Since reports of D. dendriticum in large carnivores are limited, like in the brown bear (Ursus arctos), we performed the study to investigate the presence and the prevalence of D. dendriticum in free ranging brown bears from different Croatian regions. Data on age, gender, location at the time of death, season, intensity of infection and pathohistological changes have been collected from infested animals. Parasites from bears, goat and sheep were further genotyped to investigate possible genetic difference among D. dendriticum isolates. Between February 2014 and December 2016, we sampled the complete gall bladders and liver samples of 136, hunted and dead due to other causes, brown bears across the bear range in Croatia. Each gallbladder and content was washed through the sieve to detect and count each parasite. We found that 60% of the examined brown bears were infected, ranging from 1- 103 parasites (mean=16.31, median=1) without differences in prevalence related to location, gender or age class. Seasonal distribution of Dicrocoelium infestation indicated a significantly higher percentage of infestation in autumn (mid August-December, 72%) compared to spring (February-May, 45% ; p=0.00192). Sequencing of ITS-1 of individual D. dendriticum from 16 bears, 5 sheep and one goat didn’t show any heterogeneity. COX1 sequences were identical from all bears from various regions showing differences in 7 nucleotide position with similarity of 98% with small ruminant isolates. To evaluate possible genetic variability of D. dendriticum within single bear COX1 was sequenced from 3-5 adult flukes from 10 bears without any SNP. Histopathology showed granulation of the surface of the liver and hepatomegaly in infested individuals, as well as enlarged gall bladders with firmer consistency of bile ducts surrounding the parenchyma. In the current survey we have presented evidence that bears could serve as final host of potential new “bear” genotype of D. dendriticum. This finding also suggests possible adaptation of D. dendriticum and unique life cycle of “bear” isolate.

Since ferrets (Mustela putorius furo) were first introduced into Croatia around 15 years ago, their popularity and number has risen continuously. This increase has been met by a concomitant need for additional veterinary diagnostic attention. We conducted a retrospective study on 14 cytological, 31 histological and 23 patoanatomical diagnoses made from samples obtained from 68 individuals of the Croatian ferret population over a 7 year period. Submitted samples originated from 60, 7% male and 39, 3% female animals, with an age median of 5, 1 years and an age range from 1 year to 9 years old. All cases diagnosed at the Department of Veterinary Pathology Zagreb were classified as either neoplasms (n=37), inflammatory lesions (n=15), degenerative disorders (n=10) or miscellaneous lesions (n=6). The most sensitive ferret organ systems to injurious stimuli were the haemolymphatic system, skin and endocrine system. Neoplasia presented as the major type of injury to organs with multicentric lymphoma being the leading diagnosis (diagnosed in 12 cases). Although ferrets are still a relatively rare kind of companion in Croatia, lesions described in our survey correspond to lesions in ferret populations in other countries. As Croatian ferrets are vaccinated and also kept solitary in most cases, infectious diseases were not found in our survey.