Efikasnost detoksikujućeg agensa kod T-2 toksikoze brojlera

This investigation was conducted in order to investigate the efficacy of the detoxifying agent Mycofix® Plus (MP) in the prevention and/or alleviation in vivo adverse effects of T-2 toxin in broilers. In addition, the adsorbing potential of MP was estimated in vitro. Mean degradation levels of T-2 toxin with MP in vitro, as measured by HPTLC, varied from 26.06 to 31.02 % and the adsorption ability was elevated in acidic environment (pH 3). In vivo trial was performed on 160 one day old 'Ross' broiler chicks and lasted for 21 days. Birds were divided into 4 equal groups as follows: Group 1 - negative control; Group 2 - positive control - 2 ppm T-2 toxin; Group 3 - 2 ppm T-2 toxin+2 kg/t MP; Group 4 - 2 kg/t MP. Broilers fed the diet containing 2 mg/kg of T-2 toxin without MP developed typical T-2 toxicosis. Birds that were fed the diet containing both T-2 and MP had better performances and no oral ulcerations as the dominant sign of T-2 toxicosis were observed. Histopathological examination of tissues originating from birds fed the diet containing T-2 toxin revealed degenerative changes in the oral and small intestine mucosa, necroses of enterocytes and hepatocytes, as well as depletion of lymphocytes in the bursa Fabricii. Immunohistochemical examination also revealed negative effects of T-2 toxin on cells proliferation in intestineal and bile duct mucosa, as well as on lymphocytes from bursa Fabricii. The macroscopic and microscopic structure of the liver, intestine and bursa Fabricii of broilers fed a diet containing T-2 toxin and MP was mostly preserved. Cutaneous basophile hypersensitivity reaction was weaker in broilers fed mixtures containing 2 mg/kg T-2 toxin.U cilju ispitivanja efikasnosti detoksikujućeg agensa Mycofix® Plus (MP) na preveniranje ili ublažavanje štetnih efekata T-2 toksikoze kod brojlera izvedena su ispitivanja u in vitro uslovima, kao i in vivo ogled. Prosečne vrednosti degradacije T-2 toksina od strane MP u in vitro uslovima, ustanovljene HPTLC metodom, su iznosile 26,06 do 31,02 %, pri čemu je konstatovana veća sposobnost adsorpcije ovog toksina u kiseloj sredini (pH 3). In vivo ogled je bio postavljen na 160 jednodnevnih brojlera provenijencije 'Ross', podeljenih u 4 eksperimentalne grupe sa po 40 jedinki u svakoj i trajao je 21 dan. Praćenjem zdravstvenog stanja, kliničke slike i proizvodnih rezultata, kod brojlera koji su u hrani dobijali umešan T-2 toksin u količini od 2 mg/kg, konstatovani su karakteristični simptomi za T-2 toksikozu. Kod brojlera koji su uz toksin, hranom dobijali iMP, konstatovani su zaštitni efekti u vidu izostanka ulceracija u usnoj duplji i boljih proizvodnih rezultata u odnosu na brojlere koji su dobijali samo T-2 toksin. Patohistološkim ispitivanjima uočeni su negativni efekti T-2 toksina u svim ispitivanim organima u vidu degenerativnih promena na sluzokoži usne duplje i tankog creva, nekroza enterocita i hepatocita kao i deplecija limfocita u Fabricijevoj burzi. Imunohistohemijskim ispitivanjima uočeni su negativni efekti T-2 toksina na proliferativnu aktivnost ćelija mukoze creva, žučnih kanala i limfocita u Fabricijevoj burzi. Kod brojlera koji su putem hrane dobijali T-2 toksin i adsorbens MP, zapaža se uglavnom očuvana makroskopska i mikroskopska struktura jetre, creva i Fabricijeve burze. Reakcija kožne preosetljivosti na PHA je bila slabija kod brojlera koji su hranom dobijali samo T-2 toksin

Distribucija zvezdastih ćelija jetre i njihova uloga u nastanku parazitske fibroze i ciroze jetre kod domaćih životinja

Increasing of the extracellular matrix in rats, as well as in humans, occurs as a consequence of hepatic stellate cells (HSCs) activity. The objective of this work was to investigation the role of these cells in the development of fibrosis and liver cirrhosis which occurs as a consequence of infection of sheep and goats with large (Fasciola hepatica) and small (Dicrocoelium dendriticum) fluke. Liver samples taken from 12 cattle and 10 sheep infected under natural conditions with large and small fluke were fixed in formalin and embedded in paraffin. Paraffin clips were stained with hematoxylin- eosin and masson trichrome method, and immunohistochemical method for α-smooth muscle actin (α-SMA). All tested samples were divided into three groups according to histological criteria: livers of infected animals with the first degree of fibrosis, livers of infected animals with the second degree of fibrosis, and livers of infected animals with cirrhosis. Distribution of HSCs depended on the degree of liver fibrosis. Immunohistochemically reactive HSCs were predominantly placed in perisinusoidal space. In liver samples with cirrhosis, HSCs were placed on the periphery of pseudolobulus. Cells of a different shape and size were positive to α-SMA. HSCs play an important role in synthesis of components of extracellular matrix during the development of parasitic fibrosis and liver cirrhosis in domestic animals.Povećanje ekstracelularnog matriksa kod pacova, kao i kod ljudi, nastaje kao posledica aktivacije zvezdastih ćelija jetre (HSCs). Cilj ovog rada je izučavanje uloge ovih ćelija u razvoju fibroze i ciroze jetre koje nastaju kao posledice infekcija ovaca i goveda velikim (Fasciola hepatica) i malim (Dicrocoelium dendriticum) metiljem. Uzorci jetri 12 goveda i 10 ovaca koje su u prirodinim uslovima inficirane velikim i malim metiljem su fiksirani u formalinu i uklopljeni u parafin. Parafinski isečci su bojeni hematoksilin-eozinom i Mason-trihromnom metodom, kao i imunohistohemijskom metodom za α-glatko-mišićni aktin (α-SMA). Svi ispitani uzorci su prema histološkim kriterijumima podeljeni u tri grupe: jetre inficiranih životinja sa I stepenom fibroze, jetre inficiranih životinja sa II stepenom fibroze i jetre inficiranih životinja sa cirozom. Distribucija HSCs je zavisila od stepena fibroze jetre. Imunohistohemijski reaktivne HSCs su bile predominantno smeštene u perisinusoidnim prostorima. U uzorcima jetre sa cirozom HSCs su bile smeštene na periferiji pseudolobulusa. Ćelije različitog oblika i veličine su bile pozitivne na α-SMA. HSCs igraju važnu ulogu u sintezi komponenti ekstracelularnog matriksa tokom razvoja parazitske fibroze i ciroze jetre kod domaćih životinja

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

Patomorfološke promene kod avijarne influence domaćih i divljih ptica – tehnike obdukcije ptica; pravilno uzorkovanje i slanje materijala za dijagnostiku

Usled jesenje migracije ptica ka teritoriji Evrope, postoji kontinui- rani rizik od pojave novih slučajeva visoko patogene avijarne influence (HPAI), kako kod divljih, tako i kod domaćih ptica. Tokom jeseni 2020. godine dijagnostikovano je preko 300 slučajeva HPAI-A(H5) kod divljih ptica i živine u zemljama EU. Broj žarišta HPAI se drastično povećava i dok je proteklih godina glavni uzročnik epidemije bio A(H5N8) podtip virusa, sada je zastupljeno nekoliko podtipova: A(H5N8), A(H5N5) i A(H5N1). Kod infekcije HPAI virusima, tip promena koje se sreću na obdukciji, kao i njihova lokalizacija mogu se donekle razlikovati u zavi- snosti od podtipa virusa i vrste ptica. Promene se javljaju u vidu nekro- za, edema i hemoragija, a najčešće su locirane na plućima, srcu, jetri, slezini, bubrezima, mozgu i koži. Obdukcija ptica sumnjivih na HPAI se obavlja u prostorijama sa biosigurnosnim nivoom 2, noseći, pored standardne zaštitne opreme još i maske N95. Obdukcija se vrši prema standardnim protokolima za obdukciju ptica koji uključuju spoljašnji i unutrašnji pregled leša. Prilikom obdukcije, potrebno je uzeti uzorke tkiva promenjenih organa i ona, koja su namenjena za patohistološ- ku analizu, fiksirati u 10% puferizovanom formalinu najmanje 48 ča- sova. Uzorci ne smeju biti deblji od 0,5 cm, kako bi fiksativ magao da prodre kroz tkivo. Obdukovane leševe ptica treba neškodljivo ukloniti, kao materijal kategorije 1

Ekspresija KIT receptora na kutanim mastocitomima pasa bez c-kit mutacija

Histopathological examination, grading, immunohistochemical staining and molecular genetic examinations are the proposed criteria that should be used for cutaneous mast cell tumors (CMCTs) classification. The presence of aberrant CD117 expression and mutations of the c-kit proto-oncogene could be an indicative parameter for final histological grading. Determination of the connection between the localization of KIT receptor expression and the histological grade of CMCTs without c-kit proto-oncogene mutations was the main goal of this study. The study included twenty four CMCTs and six control skin samples from 30 dogs of different ages, breed and sex. Formalinfixed and paraffin-embedded tissue samples were stained with hematoxylin-eosin and toluidine blue and immunohistochemically tested for CD117 expression. DNA was extracted from the same paraffin blocks and subsequent polymerase chain reaction amplification was performed using PE1 and PE2 primers. Degree of malignancy was determined based on the presence of mitotic figures, multinucleated cells, bizarre nuclei and karyomegaly in 10 high power fields. Based on histological features, fourteen of 24 CMCTs were of a high histological grade, while ten were classified as a lowgrade malignancy. CD117 cytoplasmic expression was observed in nine of fourteen high-grade malignancy CMCTs, which confirms the link between the aberrant CD117 expression and increased cell proliferation.Histopatološko ispitivanje, gradacija, imunohistohemijsko bojenje i molekularno genetska ispitivanja su predloženi kriterijumi za klasifikaciju kutanih mastocitoma pasa. Prisustvo aberantne ekspresije CD117 i mutacija c-kit protoonkogena moglo bi da bude indikativni parameter za finalnu histopatološku klasifikaciju mastocitoma. Glavni cilj ispitivanja je određivanje povezanosti mesta ekspresije KIT receptora i histološkog gradusa mastocitoma bez c-kit mutacija. Ispitivanjima su obuhvaćena 24 mastocitoma i 6 kontrolnih isečaka kože uzorkovanih od ukupno 30 odraslih pasa različitog uzrasta, rase i pola. Uzorci fiksirani u formalinu i ukalupljeni u parafinske kalupe su bojeni hematoksilin-eozinom, toluidin plavo metodom i imunohistohemijski primenom CD117 antitela. DNK je izolovana iz istih parafinskih kalupa, a PCR amplifikacija je izvršena primenom specifičnih prajmera PE1 i PE2. Stepen malignosti je utvrđen na osnovu prisustva mitotskih figura, multinuklearnih ćelija, ćelija sa bizarnim jedrom i kariomegalije na 10 polja velikog uveličanja. Na osnovu histopatoloških osobina, 14 od 24 mastocitoma su svrstani u visoki stepen maligniteta, dok je 10 mastocitoma niskog stepena maligniteta. Citoplazmatska ekspresija CD117 je uočena kod 9 od 14 mastocitoma visokog stepena maligniteta, što potvrđuje vezu između aberantne ekspresije CD117 i povećane ćelijske proliferacije

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

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

Wooden breast - a novel myopathy recognized in broiler chickens

Abnormally hard breast fillet consistency began to emerge in commercial broiler chickens around 2010. Due to the remarkable muscle hardness, the condition acquired the vernacular name wooden breast myopathy. This myopathy starts to develop after two weeks of age at the earliest and typically proceeds into chronic myodegeneration in three to four weeks of age. The lesion begins focally and typically develops into a diffuse lesion that involves the entire major pectoral muscle. The restricted location of wooden breast lesion in the m. pectoralis major distinguishes it from several other myodegenerative diseases that widely affect the skeletal muscle system and often the cardiac and smooth muscle systems too. Although industry-wide incidence rates are difficult to assess, it has been estimated that approximately 5-10% of commercially produced breast fillets exhibit severe WB. Even at low incidence rates, the costs to industry are substantial, as breast fillets with the wooden breast condition are often downgraded and sold at a discount, used for further processing, or in extreme cases, discarded. Because the etiology of wooden breast is still unclear, in the future, study of the early lesions, pathogenesis and the possible reduction of animal welfare are likely to gain more attention

Identifikacija optimizovanom taqman probom qpcr metodom gena rezistencije na scrapie kod ovaca na teritoriji Republike Srbije

Scrapie is an infectious neurodegenerative disease affecting the central nervous system of sheep and goats that belongs to transmissible spongiform encephalopathies. The disease is caused by the accumulation of proteinase-resistant isoform of the prion protein. The sheep predisposition to scrapie is associated with polymorphisms of the PrP gene. Genetic susceptibility to scrapie is mainly related to codons 136, 154, and 171. ARR sheep are strongly scrapie resistant and VRQ genotype is the most susceptible. Many countries have scrapie eradication programs based on using rams with resistant genotype. The eradication program has not yet been implemented in the Republic of Serbia. To examine the genetic makeup of sheep in Serbia related to scrapie, we optimized TaqMan probes of real-time polymerase chain reaction (qPCR) technique for three codons. Blood samples from 100 sheep were analyzed by qPCR and the majority of the examined sheep were AA homozygous for the 136 codon. For codon 154 the most frequent genotype was RR and for codon 171 the most frequent genotype was QQ.Ovčija svrabež je infektivno neurodegenerativno oboljenje iz grupe transmisivnih encefalopatija koje uzrokuje promene u moždanom tkivu ovaca i koza. Oboljenje je uzrokovano akumulacijom proteinaza-rezistentne izoforme prion proteina. Osetljivost ovaca na ovo oboljenje uslovljena je polimorfi zmom PrP gena. Genetska otpornost na ovčiju svrabež prevashodno je uslovljena kodonima 136, 154 i 171. Najotpornije su ovce sa ARR genotipom, dok se ovce sa VRQ genotipom smatraju najosetljivijim. Mnoge zemlje su sprovele programe eradikacije ovčije svrabeži bazirane na korišćenju priplodnih ovnova koji imaju rezistentan genotip. Ovakav program još nije sproveden u Republici Srbiji. U cilju ispitivanja genetskog statusa ovaca u Srbiji u pogledu otpornosti na ovčiju svrabež, optimizovali smo TaqMan probu, kvantitativne lančane reakcije polimeraze (qPCR) tehnike za sva tri kodona. Ispitivanjem uzoroka krvi od 100 ovaca primenom RT-PCR ustanovljeno je da je većina ovaca AA homozigot za kodon 136. Za kodon 154 najčešći genotip je RR, a za kodon 171 genotip QQ

Histopathological characteristics and expression of cdv-np antigen in the brain of serologically positive spontaneously infected red foxes (vulpes vulpes) in western Serbia

Canine distemper virus (CDV) is a worldwide distributed RNA virus that can cause severe disease in carnivore and non-carnivore species. Red foxes are highly susceptible and may act as a reservoir of the virus. As in other wild species, distemper in red foxes can manifest as acute, systemic and chronic nervous form. In the present study, we detected antibodies against CDV among red foxes in Western Serbia, and analyzed histopathologically and immunohistochemically for CDV nuclear protein antigen (CDV-NP) brain samples derived from seropositive animals. Seroprevalence of CDV antibodies was 36.8%. Histopathological changes included gliosis, neuronal degeneration, satellitosis, mononuclear inflammation, demyelination and presence of inclusion bodies. Immunostaining showed a diffuse presence of CDV-NP antigen, mainly in the cytoplasm of astrocytes and neurons. Results of this work contribute to the opinion that red foxes act as a potential reservoir of CDV and underline the importance of routine vaccination of dogs that could come in close contact with these animals. Potential active surveillance program would give a better insight in the degree of CDV infection in wildlife