Analiza morfoloških promena, ekspresije i distribucije virusnog antigena u mozgu lisica prirodno inficiranih virusom štenećaka

In this doctoral dissertation brain samples of red foxes (Vulpes vulpes) naturally infected with canine distemper virus (CDV) were examined. After exclusion of animals positive for rabies using direct immunofluorescence test, red foxes' blood serum samples were examined using enzyme-linked immunosorbent assay to detect CDV antibodies. Gross and microscopic examinations of samples of serologically positive animals were performed. The viral antigen expression and distribution in the brain of affected animals were revealed immunohistochemically, as well as the nature of perivascular inflammatory infiltrate. The degree of demyelination was visualized using luxol fast blue staining. Molecular studies were performed to detect the presence of viral RNA. Patomorphological changes were statistically evaluated. Animals positive for rabies were not detected. Presence of CDV antibodies was detected in 36.8% of examined animals with no link between the antibody titer and the amount of pathological changes. Although no gross changes were observed, numerous changes consistent with non-purulent panencephalitis and demyelinating leukoencephalitis were evident microscopically. Immunohistochemical staining revealed moderate to strong positive reaction against viral nucleoprotein (CDV-NP), as well as the presence of T cells in the perivascular inflammatory infiltrate. Demyelination was the most pronounced in the cerebellar white matter. Using the molecular method (RT-PCR) viral RNA was not detected. Statistical evaluation, comparing the mean values of histopathological changes, showed a significant difference between various brain segments.U ovoj doktorskoj disertaciji ispitivani su uzorci mozga crvenih lisica (Vulpes vulpes) prirodno inficiranih virusom štenećaka. Nakon isključivanja životinja pozitivnih na besnilo metodom direktne imunofluorescencije, vršena su ispitivanja uzoraka seruma crvenih lisica imunoenzimskom metodom u cilju utvrđivanja prisustva antitela protiv virusa štenećaka. Rađena su makroskopska i mikroskopska ispitivanja uzoraka mozga serološki pozitivnih jedinki. Ekspresija i distribucija virusnog antigena u mozgu inficiranih lisica utvrđena je imunohistohemijski, kao i ispitivanje prirode inflamatornog ćelijskog infiltrata. Histohemijskim bojenjem LFB metodom određen je stepen demijelinizacije bele moždane mase. Molekularno-genetička ispitivanja korišćena su za dokazivanje virusne RNK. Izvršena je statistička obrada rezultata utvrđenih patomorfoloških promena. Nije utvrđeno prisustvo jedinki pozitivnih na besnilo. Prisustvo antitela utvrđeno je kod 36,8% životinja i nije utvrđen uticaj visine titra antitela na izraženost patomorfoloških promena. Iako nisu utvrđene makroskopske promene, mikroskopski su se uočavale brojne promene u tipu negnojnog panencefalitisa i demijelinizujućeg leukoencefalitisa. Imunohistohemijski, utvrđena je pozitivna reakcija umerenog do jakog intenziteta protiv virusnog nukleoproteina (CDV-NP), kao i prisustvo T ćelija u perivaskularnom infiltratu. Demijelinizacija je bila najizraženija u beloj masi malog mozga. Upotrebom molekularno-genetičke metode (RT-PCR) nije utvrđeno prisustvo virusne RNK. Poređenjem srednjih vrednosti histopatoloških promena statističkom obradom podataka utvrđena je signifikantna razlika između pojedinih segmenata mozga

Morphological characteristics of three classic forms of natural equine glanders - a disease with high zoonotic significance

Equine glanders is a severe bacterial disease known since ancient times. Although eradicated in the most part of the world it is now considered re-emerging. Considering very scarce literature data, we used from formalin fixed collection material: nasal septum, lung and skin specimens from naturally infected horses. Tissues were grossly examined and photographed. Tissue samples, after standard processing, were stained with HE, Congo red and Groccot and microscopically examined. Gross changes include nodules and ulcers in the nasal mucosa with granulation and scarring, pyogranulomas in the lung tissue and nodules and ulcers of the skin. Microscopically marked inflammation of affected tissues with neutrophilic and mononuclear infiltration and fibrous tissue proliferation were seen. As a potent zoonotic agent it has been already used as a biological weapon in the past

Mukometra kao posledica ozbiljne stručne greške

An apathetic 14-year-old Hungarian Puli bitch was presented with a seven day history of vaginal discharge and reported ovariohysterectomy (OHE) conducted six years ago. Clinical examination showed elevated body temperature (39.5°C), enlarged abdomen, and vulvar swelling. Despite a reported OHE in the anamnesis, ultrasound examination of the abdomen demonstrated the presence of both ovaries and a large anechogenic zone resembling the uterus. Vaginal cytologic examination and hormonal analysis indicated that the bitch was in the oestrus phase of the cycle. After laparotomy, both intact ovaries and an enlarged uterus with marked adhesions and filled with liquid content were visualised. The cervix was situated in the caudal part of the abdominal cavity and connected to the uterus by only an adhesion stump. After extirpation, uterus and ovarian tissue samples were sent for histological examination. Histological findings of uterus and ovarian tissue samples indicated a diagnosis of mucometra. Obviously, OHE was not conducted professionally, and the procedure performed only prevented conception, but ovaries and uterus continued to be active and caused a serious health disorder after six years. When only a part of the ovaries or uterus is left after OHE, health complications can appear up to ten years later. Our case testifies that even if both ovaries and the entire uterus are left after OHE without communication with cervix and vagina, a bitch can live without noticeable health disorders up to six years. This underlines the importance of lymphatic drainage and resorption processes in the uterus as well as evacuation of uterus content through the vagina.Četrnaest godina stara kuja rase mađarski puli je dovedena u apatičnom stanju i sa anamnezom iscedka iz vagine koji je prisutan 7 dana. Prema rečima vlasnika ovariohisterektomija je urađena pre 6 godina. Kliničkim nalazom je utvrđena povišena telesna temperatura (39,5˚C) uvećan abdomen i otok vulve. Uprkos podacima iz anamneze ultrazvučni pregled je ukazivao na prisustvo oba jajnika i velike anehogene zone koja je ukazivala na prisustvo materice. Citološki nalaz brisa vagine i hormonska analiza su ukazivali na to da je kuja bila u estralnoj fazi ciklusa. Nakon laparotomije potvrđeno je prisustvo oba jajnika i uvećane materice koja je bila ispunjena tečnim sadržajem i sa velikim brojem adhezija. Cerviks je bio lociran u kaudalnom delu abdomena i pričvršćen za uterus samo adhezionim patrljkom. Nakon ekstirpacije uterus i ovarijumi su poslati na histološku analizu, kojom je utvrđena dijagnoza mukometre. Očigledno da prethodno urađena ovariohisterektomija nije bila obavljena stručno i da je urađeni postupak sprečio koncepciju ali su ovarijumi i materica nastavili da budu aktivni što je prouzrokovalo ozbiljan zdravstveni problem 6 godina kasnije. Kada nakon ovariohisterektomije ostanu samo delovi tkiva ovarijuma i uterusa zdravstvene komplikacije se mogu javiti i do 10 godina nakon operacije. Prikazani slučaj svedoči o tome da je, čak i u slučaju kada su nakon operacije ostavljeni oba ovarijuma i uterus bez komunikacije sa cerviksom i vaginom, kuja 6 godina živela bez ozbiljnijih zdravstvenih problema. Ovo naglašava značaj limfne drenaže i procesa resorpcije sadržaja iz materice, pored pražnjenja kroz cerviks i vaginu

Morphological characteristics and expression of estrogen and progesterone receptors in the canine endometrium during the estrus cycle, cystic endometrial hyperplasia and pyometra

The estrus cycle of bitches is divided into four phases: proestrus, estrus, diestrus and anestrus, during which different morphological changes, and also cyclic changes of estrogen and progesterone receptors are present. Several pathological changes can be differentiated on the endometrium, but one of these is the most important - cystic endometrial hyperplasia, which frequently develops into pyometra. The aim of the present study was to describe morphological characteristics, and expression of estrogen and progesterone receptors on the endometrium of mixed-breed bitches during the different phases of the estrus cycle, cystic endometrial hyperplasia and pyometra. The uterus and ovaries of 36 mixed breed bitches in different phases of the estrus cycle and also with cystic endometrial hyperplasia (CEH) and chronic purulent endometritis - pyometra were examined macroscopically, histopathologically, and immunohistochemically for estrogen receptors (ER) and progesterone receptors (PR). During proestrus uterine cells showed a weak reaction for both estrogen and progesterone receptors, but during estrus a large number of uterine cells showed a strong reaction on estrogen receptors and moderate reaction on progesterone receptors. On the contrary, during diestrus the scores for the estrogen receptors decreased, while the progesterone receptors level increased - uterine cells expressed strong reaction for progesterone receptors, and moderate reaction for estrogen receptors. Uterine cells in cystic endometrial hyperplasia expressed a strong reaction for estrogen receptors, and moderate reaction for progesterone receptors, but on the other hand the uterine cells in the uterus with pyometra expressed a moderate to strong reaction for progesterone receptors, and a weak reaction for estrogen receptors. In further investigations it would be interesting to perform quantitative analysis for both estrogen and progesterone receptors during different phases of the estrus cycle and also in the uterus with cystic endometrial hyperplasia and pyometra

Morphological features and kit receptor expression in canine cutaneous mast cell tumor and systemic mastocytosis

Mast cell neoplasia in dogs can occur in two different forms: common as cutaneous tumor, or less common as a systemic form of neoplastic mast cell proliferation - systemic mastocytosis. The aim of this study was to compare the histological and cytological features, KIT receptor expression and presence of c-KIT proto-oncogene mutations in neoplastic cells of dogs with canine cutaneous mast cell tumor (CMCT) and systemic mastocytosis. Microscopical examination of the cytological smears obtained from all selected dogs revealed that cellular specimens were constituted mostly of round cells with a central nuclei and fine to coarse purple cytoplasmic granules. Histopathological examination of skin samples of dogs with CMCT and a dog with systemic mastocytosis showed proliferation of the neoplastic mast cells in the superficial and/or deep dermis. Similar findings were observed in tissue samples derived from lymph nodes, spleen, liver, myocardium and kidneys of a dog with systemic mastocytosis. Three dogs with high grade CMCT as well as one dog with systemic mastocytosis showed cytoplasmic CD117 expression, while 3 dogs with low grade CMCT, had membranous expression of CD117. Based on our study, histological features and cytoplasmic CD117 expression in neoplastic cells of dogs with systemic mastocytosis are similar to those in dogs with high grade CMCTs. Nevertheless, mutations of c-KIT proto-oncogene were not found in tumor samples either from dogs with CMCT or dog with systemic mastocytosis

Patohistološki i imunohistohemijski parametri značajni za prognozu i terapiju tumora pasa i mačaka

The development of tumours implies a disproportion between proliferation, growth and differentiation, which is accompanied by significant genotypic and phenotypic variations in the diseased. The scenario of multiple oncogenic effects on the inhibition of cell growth, apoptosis and maturation, on the stimulation of proliferation, on cell migration and tissue invasion, is responsible for the occurrence of tumours both in humans and in ani­mals. The qualitative and quantitative determination of biological prognostic factors in the tissue of affected cats and dogs directly indicates a possible tumour metastasizing, and consequently also the prognosis of neoplastic disease. Monitoring of clinical and biological prognostic factors is important for therapy and prediction of tumours in humans and animals. The determination of the proliferative potential of tumour cells by immunohistochemical detection of Ki-67 and PCNA is used in grading certain tumours in cats and dogs. It is also possible immunohistochemically to prove factors of angiogenesis and numerous activating and inhibiting proteins in the tumour tissue. In addition to other histological, clinical and biological parameters, they are significant for the further prognosis and therapy of neoplastic diseases.Razvoj tumora podrazumeva nesklad između proliferacije, rasta i diferencijacije, što je praćeno značajnim genotipskim i fenotipskim varijacijama kod obolele jedinke. Brojni onkogeni efekti inhibišu starenje ćelije, apoptozu i maturaciju, a stimulišu proliferaciju, ćelijsku migraciju i tkivnu invaziju kod ljudi i životinja. Kvalitativno i kvantitativno određivanje bioloških prognostičkih faktora u tkivu obolelih pasa i mačaka govori direktno o sposobnosti metastaziranja tumora, a time i o prognozi neoplastične bolesti. Praćenje kliničkih i bioloških prognostičkih faktora značajno je za terapiju i predikciju tumora kod ljudi i životinja. Određivanje proliferativnog potencijala tumorskih ćelija, imunohistohemijskom detekcijom Ki-67 i PCNA koristi se u gradiranju nekih tumora kod pasa i mačaka. Faktore angiogeneze i brojne aktivirajuće i inhibitorne proteine, moguće je takođe imunohistohemijski dokazati u tkivu tumora. Oni su uz ostale histološke, kliničke i biološke parametre značajni za dalju prognozu i terapiju neoplastičnih bolesti

Mastocitom unutar lipoma kod bele toj pudle

Uprkos činjenici da su mastocitomi jedna od najčešćih neoplazmi kod pasa, postoji samo jedan objavljeni slučaj mastocitoma u lipomu. U ovom slučaju, kod 11-godišnje ženke toj pudle primećeno je prisustvo dobro ograničene, pokretne, meko-elastične mase na lateralnoj strani grudnog koša. Histopatološki pregled je pokazao dobro ograničenu masu koja se sastojala od zrelih adipocita između kojih se nalaze skupine ćelija sa granuliranom citoplazmom. Bojenjem toluidin plavim je dokazano prisustvo metahromatskih granula u citoplazmi mastocita. Histopatološke i imunohistohemijske karakteristike ukazale su na to da je ispitani mastocitom niskog stepena maligniteta

Morphological characteristics of skin lesions in cattle naturally infected with lumpy skin disease virus in Serbia

Lumpy skin disease (LSD) is a disease of cattle and domestic buffaloes caused by Capripoxvirus which can lead to significant economic losses. Until several years ago it was limited to Africa and the Middle East, but recently it was reported in Turkey (2013), Greece (2015), and in 2016 it spread to Bulgaria, North Macedonia, Albania, Montenegro and Serbia. The aim of this study was to determine the microscopic lesions in the skin of naturally infected animals, highlight their features and compare them to the findings of other authors. Gross lesions in the skin of cattle infected with Lumpy skin disease virus (LSDV) were manifested in the form of skin nodules which were round, raised, clearly circumscribed, firm and randomly distributed in the whole skin including the vulva and udder. Histopathological lesions included degeneration of epithelial cells, hyperkeratosis, spongiosis, and acanthosis present in 85.88%, 81.18%, 84.71%, and 80.0% of samples, respectively. Intracytoplasmic inclusion bodies were noted in keratinocytes of the stratum spinosum in 56.47% of samples, and in hair follicle cells and sebaceous glands epithelial cells in 45.88% of samples. Cutaneous lesions were in the form of inflammatory infiltrate present in the dermis and subcutis in 97.65% of samples. Since there are only a few literature data in this field, this study expands the knowledge relating to morphological alterations in LSD. Based on the characteristic microscopic lesions described in the present study, histopathology should be considered as a very useful method for the diagnosis of LSD

The frequency of foreign bodies in the lower respiratory tract in the regoin of Slavonski Brod

Retrospektivno je analizirana učestalost stranih tijela donjih dišnih putova, u periodu od 1961. do 1987. godine. U tom periodu 81 osoba je aspirirala strana tijela u donje dišne putove, a muškarci čine dvije trećine slučajeva. Djeca do pete godine učestvovala su u 77,8 %, a vegetabilna strana tijela su nađena u 81,5 % slučajeva. U prvih 24 sata ekstrahirana su 80,3%. U larinksu je zaostalo 8,6%, a u traheji 24,7% stranih tijela. Multiplih je nađeno u 13,6% slučajeva. Peroralnom bronhoskopijom odstranjeno je 79,7 %, a traheotomijom i donjom bronhoskopijom 20,3 % stranih tijela. Smrtnost pri ekstrakciji iznosila je 1,3 %, a sedam osoba se ugušilo na putu do bolnice (8,6 %). Godišnja incidencija na 100.000 stanovnika iznosi 2, a mortalitet 0,2.The frequency of foreign bodies in the lower respiratory tract was analyzed for the period 1961 —1987. In that period 81 persons aspirated foreign bodies into the lower respiratory tract. Two thirds of the cases were males, and 77.8 % were children up to the age of five. Vegetable foreign bodies were found in 81,5 % of cases, and 80,3 % of foreign bodies were extracted within the first 24 hours. Foreign bodies were localized in larynx in 8,6 % and in trachea in 24,7 % of cases. Multiple localizations were present in 13,6 % of cases. Foreign bodies were extracted by bronchoscopy in 79,7 % of cases and by tracheotomy and lower bronchoscopy in 20,3 %. Mortality rate during extraction was 1,3%, whereas 7 persons choked on the way to thehospital (8,6%). The annual incidence of foreign bodies in the lower respiratory tract is 0.02 %o and the mortality is 0.002 %o

Immunohistochemical distinguishing between canine peripheral nerve sheath tumors and perivascular wall tumors

Peripheral nerve sheath tumors (PNSTs) comprise a heterogeneous group of neoplasms originating from the elements of the nerve sheath. They are divided into two forms: benign and malignant PNST. Both benign and malignant PNSTs are not very common in domestic animals but they are reported in different animal species. Histologically, PNSTs are composed predominantly of spindle cells arranged in bundles, whorls and sheets, with a different number of pleomorphic cells and mitotic figures. The aim of this study was a reclassification of 17 dog tumor samples initially diagnosed with peripheral nerve sheath tumors using histopathological analysis. The main criterion for reclassification was immunohistochemical positivity for various antigens. PNSTs are often histologically very similar to other spindle cell tumors and immunohistochemistry is required for differential diagnosis. PNSTs generally express vimentin, S-100 protein, glial fibrillary acidic protein (GFAP), collagen IV and laminin. Four tumor samples were positive to muscular marker alpha-SMA and vimentin and negative for S-100 protein and desmin. The spindle cells whirling around some blood vessels were observed in these tumors so they were reclassified as perivascular wall tumors (PWTs). The other 13 tumors were S-100 protein and vimentin positive and alpha-SMA and desmin negative, thus classified as PNST. The use of the immunohistochemical panel is necessary for distinguishing PNSTs from PWTs in routine diagnostics